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Effects of monensin in transition Holstein dairy cows fed diets containing citrus pulp

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Effects of monensin in transition Holstein dairy cows fed diets containing citrus pulp
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Melendez, Pedro G., 1966-
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English
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xiii, 193 leaves : ill. ; 29 cm.

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Calcium ( jstor )
Calving ( jstor )
Dairy cattle ( jstor )
Ketosis ( jstor )
Lactation ( jstor )
Milk ( jstor )
Milk production ( jstor )
Milk yield ( jstor )
Propionates ( jstor )
Rumen ( jstor )
City of Gainesville ( local )
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bibliography ( marcgt )
theses ( marcgt )
non-fiction ( marcgt )

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Thesis:
Thesis (Ph. D.)--University of Florida, 2004.
Bibliography:
Includes bibliographical references.
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Printout.
General Note:
Vita.
Statement of Responsibility:
by Pedro G. Melendez.

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EFFECTS OF MONENSIN IN TRANSITION HOLSTEIN DAIR COWS FED DIETS
CONTAINING CITRUS PULP





By

PEDRO G. MELENDEZ






















A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE
UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY

UNIVERSITY OF FLORIDA

2004














To Oscar and Eliana, my parents,



I wish to dedicate this dissertation to those who guided my life. They have

been suffering for being far away from my kids, my wife and my-self. Tears, efforts,

and no resting have characterized their lives; but they should be very proud for having

a son who at this moment has become faculty of one of the most prestigious Colleges

of Veterinary Medicine of the USA.

Dad and Mom, this is not mine, this is yours. Without you I would have been

nothing. I love you so much ard please pardon me for not being with you.














ACKNOWLEDGMENTS

I wish to express my sincere gratitude to:

Dr. Art Donovan (my adviser and friend) for his excellent advice, support and

enthusiasm on my project and my entire graduate program at UF.

Dr. Jesse Goff, distinguished USDA researcher, for accepting to be part of my

committee, for his expertise and friendship.

Dr. Carlos Risco, Professor of Veterinary Medicine for his valuable comments

and help and for his friendship.

Dr. Ramon Littell, Professor of Statistics, IFAS, for accepting to be part of my

committee and for his valuable help on the statistical approach.

Dr. Louis Archbald, Professor of Veterinary Medicine and LACS graduate

coordinator, for accepting to be part of my committee and for his advice on my entire

program.

Dr. Charles Courtney and the Graduate School of the College of Veterinary

Medicine, UF.

The Department of Large Animal Clinical Science and Dr. Eleanor Green

Pharmacia Animal Health for their financial support.

Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile.

Mr. Don Bennink, Dr. Jeniffer McHale, Bobby, and the staff of North Florida

Holsteins Dairy Farm.



iii








ELANCO Animal Health, USA and Mr. Leo Richardson and Richard Tucker.

Dr. Todd Duffield for his comments and for sending me a copy of his

dissertation.

Dr. Julian Bartolome, for his unconditional friendship.

Dr. Victor Monterroso, for his valuable comments, loyalty, and support during

the most difficult part of my staying in USA.

Rural Animal Medicine Service, Dr. Owen Rae, chairman of RAMS, Delores,

residents, internships and OPS students for their help.

Shelly Lanhart, Chris Sissle and Christina Herejk for their help on my program

and the lab analysis.

Friends from different countries for their friendship (Gina, Antonio, Martin,

Billy, etc).

My friends in Chile for their continuous contact with me during this time and

their friendship.

My brother, grandmother, nieces and nephew for their support and feelings

about us.

My father, Oscar and my mother, Eliana, for their entire life of efforts, and

affection.

My wife, Maria Ester, for her support and love and for being a wonderful and

great woman, mother and wife.

Ignacio, Diego and Elisa, for their love and for being the most important reason

to be here.





iv














TABLE OF CONTENTS

page

ACKNOWLEDGMENTS ................... ......... .................................. ........ iii

A B STR A C T ........................................................................................ xii

CHAPTER

1 INTRODUCTION.................................................... ......................1

2 LITERATURE REVIEW .............................................. .................4

The Transition Period......................................................................4
Physiological and Metabolic Changes during the Transition
P eriod .......................................... .................................. ............. .5
Dry Matter Intake..................................................................5..
Glucose and Lipid Metabolism....................................................6
Calving-Related Disorders................................................................ 8
Milk Fever, Parturient Paresis, Hypocalcemia............................... 15
Retained Fetal Membranes-Metritis Complex.............................. .. 23
Abomasal Disorders............................................................... 30
Mastitis............... ...... .................................................37
Lameness.............................. .......................................... 38
C ulling.................................................... ......... ........ ... ..... ..40
Ketosis-Fatty Liver Complex .....................................................42
M onensin .................................................. ...... ....... .............. .......59
Rumen Microbial Ecosystem.....................................................60
Volatile Fatty Acids................................................................65
Monensin and Rumen Fermentation ..............................................65
Citrus Pulp and Pectin Fermentation .............................................68
Monensin, Calving-Related Disorders, Reproductive
and Productive Responses........................................................ 69
Transition Cow Feed Management........................................................77
Energy................................................................... ....78
Protein ............................................................. ... .. ............ 80
Minerals and Vitamins..............................................................82
Dry Cow Feed Management and Body Condition Score.................................85
Fresh Cow Feed Management and Body Condition Score..............................86




v








3 ST U D Y 1......................................................................................91
Introduction ....................................................................................91
Material and Methods......................................................................92
R esults ..................................................... ............ .................. ...97
D iscussion.................... ................ .......................... ......................98


4 STUDY 2...................................................................................118
Introduction .......... ............................................................118
Material and Methods...................................................................119
Results and Discussion...................................................................123

5 STU D Y 3................................................................................ ... 140
Introduction....................................................................... ......140
Material and Methods....................................................................141
Results and Discussion ...................................................................144


6 STUDY 4 .................................. ........ .......... ........... ...............155
Introduction ........................................................ ............ ......... 155
Material and Methods ....................................................................156
Results and Discussion....................................................................160


REFERENCE LIST...............................................................................169

BIBLIOGRAPHICAL SKETCH...............................................................192
























vi













LIST OF TABLES


Table page

2-1. Predicted changes in dry matter intake for Holstein dairy
cows during the last three weeks prior to calving...................................... 5

2-2. Effect of hormones on carbohydrates and lipids metabolites
on dairy cattle............................................................................. ... 10

2-3. Summary of relationships among Calving-Related Disorders ....................... 11

2-4. Case definition, incidence and economic losses of
calving-related disorders..................................................................14

2-5. Ketone body concentrations for clinical and subclinical
ketosis according to different authors...................................................56

2-6. Ketone body field test comparison..................................................... 57

2-7. Relative volumes and number of microbial organisms.................................60

2-8. Susceptibility and resistance of ruminal bacteria to monensin....................67

2-9. Minimum requirements for dry, prepartum and fresh cows........................88

2-10. Minimum trace mineral and vitamins requirements for dry,
prepartum and fresh cows.................................................................89

2-11. Target Body Condition Scores (BCS) Scale 1-5.......................................90

3-1. Diet composition dry cow far-off, dry cow transition and lactating
Transition cow s........................................................................... ...105

3-2. Nutrient content dry cow far-off, dry-cow transition and lactating
transition diets..................................................................................106

3-3. Lactational Incidence of Calving-Related Disorders ......................................107

3-4. Summary of Logistic Regression Modeling for CRD................................... 108



vii








3-5. Reproductive responses for cows treated with or without monensin.....................109

4-1. Diet composition dry cow far-off, dry cow transition and lactating
transition cow s................................................................................ 127

4-2. Nutrient content dry cow far-off, dry-cow transition
and lactating transition diets .........................................................................128

4-3. Least Squares Means S.E.M. for rumen pH, acetate, propionate,
butyrate, valerate, total VFA, ammonia, D and L-lactate and
Acetate:Propionate by time in primiparous treated and control cows
at 10 d pp ................................................. .................................... 129

4-4. Least Squares Means S.E.M. for rumen pH, acetate, propionate,
butyrate, valerate, total VFA, ammonia, D and L-lactate and
Acetate:Propionate by time in multiparous treated and control cows
at 10 d pp.......................................................................... ............130

4-5. Least Squares Means S.E.M. for serum B-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling after
a.m. feeding in primiparous treated and control cows at 10 d pp................... 131

4-6. Least Squaresd Means S.E.M. for serum B-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling after
a.m. feeding in multiparous treated and control cows at 10 d pp.....................132

4-7. Least Squares Means S.E.M. for serum B-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling in
primiparous treated and control cows between dry-off and 21 d pp.................33

4-8. Least Squares Means S.E.M. for serum B-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling in
multiparous treated and control cows between dry-off and 21 d pp.................34

5-1. Diet composition dry cow far-off, dry cow transition and lactating
Transition cows..................................................................... .......147

5-2. Nutrient content dry cow far-off, dry-cow transition and lactating
transition diets...................................................................................148

5-3. Least Squares Means S.E.M. of ME305 and Real Milk Yield
(kg/lactation) by treatment and parity.......................................................49

6-1. Diet composition dry cow far-off, dry cow transition and lactating
Transition cow s............................................................................. 163

6-2. Nutrient content dry cow far-off, dry-cow transition and lactating


viii








transition diets............... ....................................................... .......164

6-3. Receiver-operating characteristic (ROC) analysis for milk BHB
at three different levels of serum BHB (gold standard)................................... 65

6-4. Sensitivity, specificity, positive predictive value, negative
predictive value of milk BHB test based on a serum BHB
cut-off value of 1.0 mmol/L..........................................................166

6-5. Summary of logistic regression modeling...................................................167










































ix













LIST OF FIGURES


Figure page

2-1. Pathways of glucose, amino acids and fatty acids
metabolism intersect at the citric acid cycle...............................................58

2-2. Estimated energy density required in diets of transition
cows to meet requirements for maintenance and gestation.......................... 79

2-3. Estimated percentage crude protein required in diets of transition
cows to meet requirements for maintenance and gestation...........................81

3-1. Body Condition Score by treatments at assignment and at
calving................................................... ...................... .............. 110

3-2. Daily milk yield up to 20 dpp in first lactation cows by
treatm ent............................................................................ ..111

3-3. Daily milk yield up-to 20 d pp in third lactation cows
or older by treatment............................................................... ...112

3-4. Test day milk yield by parity 1 and 2....................................................,113

3-5. Test day milk yield by parity 3 or more................................................ 114

3-6. Test day Fat % by parity 3 or more.........................................................115

3-7. Test day Protein % by parity 3 or more.................................................... 116

3-8. Survival curve for risk of non pregnancy.................................................17

4-1. BCS at dry-off and at calving in treated and control
group by parity............................................................................ ...135

4-2. Glucose, BHB, NEFA concentration in monensin and
control primiparous cows by time after feeding at 10 d pp ...........................136

4-3. Glucose, BHB, NEFA concentration in monensin and
control multiparous cows by time after feeding at 10 d pp.............................137



x








4-4. Glucose, BHB, NEFA concentration in monensin and control
primiparous cows by time between dry-off and 21 d pp.......................................138


4-5. Glucose, BHB, NEFA concentration in monensin and control
multiparous cows by time between dry-off and 21 d pp......................139

5-1. Body Condition Score by treatments and by parity at assignment
and at calving............................................................................... ... 150

5-2. Test day milk production by treatment in primiparous cows..........................151

5-3. Test day milk production by treatment in multiparous cows.............................152

5-4. Glucose, BHB, NEFA concentration in monensin and control
primiparous cows by time between dry-off and 21 d pp.......................................153

5-5. Glucose, BHB, NEFA concentration in monensin and control
multiparous cows by time between dry-off and 21 d pp...............................154

6-1. ROC curve for milk BHB for the detection of subclinical
ketosis under a gold standard of serum BHB > 1.0 mmol/L.............................168





























XI












Abstract of Dissertation Presented to the Graduate School of the University of Florida in
Partial Fulfillment of the Requirements for the Degree of
Doctor of Philosophy

EFFECTS OF MONENSIN IN TRANSITION HOLSTEIN DAIRY COWS FED DIETS
CONTAINING CITRUS PULP

By

Pedro G. Melendez

May 2004

The objective of this research was to determine the effect of monensin-controlled

release capsule on the incidence of periparturient disorders, cow performance, and blood

metabolites in Holstein cows fed diets containing citrus pulp. Four studies were con-

ducted on a dairy farm located in Florida. In all four studies, cow were assigned

randomly at dryoff (50 to 70 days before expected parturition [BEP]) to one of two

groups. The treated group received orally a monensin capsule (CRC Rumensin,

ELANCO Animal Health, Guelph, ON) that releases 300 mg of monensin per day for 95

days. Control cows were matched by parity and received no treatment. Studies 1, 2, and 3

began between July and August 2001. In study 1, 580 cows were assigned to two groups.

Outcome variables were incidence of periparturient disorders, milk yield and solids, body

condition score (BCS) at calving, and reproductive responses. In study 2, 60 cows were

randomly assigned to two groups. Outcome variables were milk yield and BCS at

calving. Blood samples were collected at assignment; 21 d BEP; at calving; and at 7, 14,

and 21 d after calving. In study 3, 300 cows were assigned similarly. At 14 days

postpartum, a milk sample was obtained for beta hydroxyl butyrate (BHB). In a


xii








subsample of 50 cows per group, a blood sample for BHB determination was taken. In

study 4, 24 cows (in March 2003) were assigned similarly. A blood sample was obtained

and BCS was conducted at assignment; on day 21 BEO; at calving; and at 7, 14, and 21 d

postpartum. A rumen fluid and blood sampling scheme was carried out on day 10

postpartum. The first sample was obtained before the morning meal. Three more samples

were taken every 2 hours. Outcome variables in rumen samples were pH; and

concentrations of NH3, acetic, propionic, butyric, and L and D-lactic acids. Outcome

variables in blood samples for studies 2 and 4 were non-esterified fatty acids (NEFA),

BHB, and glucose.

In study 1, monensin improved milk yield, decreased the incidence of metritis and

increased the incidence of dystocia in third and older parity. Milk solids were decreased

by treatment. In study 2, monensin increased BCS at calving in multiparous cows. In

study 3, monensin decreased the proportion of cows with subclinical ketosis. In study 4,

monensin decreased the level of BHB and NEFA; and increased glucose levels. Minor

changes in rumen fermentation were detected. It is concluded that monensin improved

slightly milk production and metabolism performance of transition dairy cows fed diets

containing citrus pulp.
















xiii













CHAPTER 1
INTRODUCTION

The transition period, defined as the last 3 weeks before parturition to 3 weeks after

parturition is the most stressful and challenging stage of the lactation cycle (Grummer,

1995; Drackley, 1999). During the last 10 years, research on this topic have been copious,

and for first time the National Reseach Council (NRC, 2001) has included a chapter

covering the most important and unique aspects of transition dairy cows. Since most of the

metabolic disorders of dairy cows occur during this time (Goff and Horst, 1997b;

Drackley, 1999), strategies to prevent calving-related disorders have been focused on the

nutritional and feeding management aspects of the prepartum transition dairy cow (Goff

and Horst, 1997b; Gerloff, 2000; Oetzel, 2000).

The use of feed additives has become a common practice in dairy cattle. They

improve palatability, enhance rumen fermentation, optimize rumen digestion of the fiber,

reduce mobilization of adipose tissue, and maintain calcium and phosphorus homeostasis

during the first few days of lactation (Santos, 1999). However, fed additives do not replace

the optimum levels of nutrients, but complement the effect of a good dietary management.

Unbalanced diets or poor management will not be corrected by the use of additives

(Gerloff, 2000).

Sodium monensin is an additive ionophore antibiotic that selectively modifies the

ruminal flora and improves the digestive efficiency in cattle, increasing productive

parameters and decreasing health problems. Monensin increases ruminal propionate



1





2

production, reduces production of methane and has a sparing effect on ruminal protein

digestion (Richardson et al., 1976; Yang and Russell, 1993; Nagaraja et al., 1997; Becket et

al., 1998; Duffield et al., 1998a). These metabolic changes have been related with a decrease

in the incidence of some metabolic disease such as ketosis and fatty liver and an increase in

milk yield. Monensin, used as a controlled-release capsule, during the prepartum period,

decreased the acetate to propionate ratio, increased the concentration of blood glucose,

decreased serum NEFA concentrations, increased ruminal pH, decreased the ruminal butyrate

production and subsequently decreased the BHB concentration (Duffield et al. 1998a,1998b;

Green et al., 1999). Most of these studies were conducted in Canada and other countries,

demonstrating an improvement of the overall performance of lactating dairy cows.

Unfortunate, monensin is not allowed in lactating dairy cattle in the United States.

Citrus pulp is an energy concentrate by-product produced in subtropical regions, of

which south central Florida remain the largest area of production. Citrus pulp is a common

by-product used in diets for dairy and beef cattle in Florida and other southern states

(Arthington et al., 2002). Citrus pulp is composed mostly by pectin, which is indigested by

mammalian enzymes, but can be rapidly fermented by ruminal microbes (Hall, 1997).

Diets based on citrus pulp consistently demonstrated an increase in milk fat content and

milk urea nitrogen, but not in milk yield, when compared with diets richer in starch

(Belibasakis and Tsirgogianni, 1996; Leiva et al., 2000).

Pectin-fermenting bacteria are gram-negative monensin-resisting bacteria (Nagaraja

et al., 1997; Stewart et al., 1997). The positive effect of monensin has been demonstrated

on diets rich in starch (Nagaraja et al., 1997). However, no trials have been conducted to

test the effect of monensin on transition cows fed diets based on citrus pulp. Since





3

monensin does not affect pectin-fermenting bacteria, it is reasonable to assume that dairy

cows fed monensin and diets rich in citrus pulp should not modify extensively their

pectin rumen fermentation. In addition, monensin still would improve starch and other

soluble carbohydrate rumen fermentation, with the consequent increase in rumen

propionate and blood glucose and decrease in ketone body formation and fat

mobilization.

Therefore, the objectives of this research program were

* To determine the effect of a monensin controlled-release capsule inserted at dry-off
on the incidence of calving-related disorders, milk production and reproductive
responses in transition dairy cows fed diets containing citrus pulp.
* To establish the effect of a monensin controlled-release capsule inserted at dry-off on
rumen metabolites and blood energy-related metabolites in transition dairy cows fed
diets containing citrus pulp.
To evaluate a milk ketone body test strip and to determine the incidence of subclinical
ketosis at 14 days postpartum in transition dairy cows fed diets containing citrus pulp
and that received a bolus of monensin at dry-off.












CHAPTER 2
LITERATURE REVIEW

The Transition Period of Dairy Cows

The transition period in dairy cows is defined as the last three weeks before

parturition to three weeks after parturition (Grummer, 1995). It is characterized by

tremendous metabolic and endocrine adjustments that the cows must experience from late

gestation to early lactation (Drackley et al., 2001). Perhaps the most important

physiological change occurring during this period is the decrease in dry matter intake

around parturition and the sudden increase in nutrients that cows need for milk production

(Drackley, 1999; Ingvartsen and Andersen 2000). As a result of these remarkable changes,

most of the infectious diseases and metabolic disorders occur during this time (Goff and

Horst, 1997b; Drackley, 1999). Milk fever, ketosis, retained fetal membranes (RFM),

metritis and displacement of the abomasum (DA) primarily affect cows within the first

two weeks of lactation (Drackley, 1999). Physical and metabolic stresses of pregnancy,

calving and lactation contribute to the decrease in host resistance during the periparturient

period (Mallard et al., 1998). During two weeks before and after parturition the T-cells

populations exhibit a significant decline, which contribute to the immunosuppession in

dairy cows at calving (Kimura et al., 1999). This immunosuppression during the

periparturient period leads to increased susceptibility to mastitis and other infectious

diseases (Mallard et al., 1998). Other diseases that are not clinically apparent during the

first two weeks of lactation (laminitis, ovarian cysts, endometritis) can be traced back to

insults that occurred during early lactation (Goff and Horst, 1997b).


4





5

Physiological and Metabolic Changes during the Transition Period

Dry Matter Intake

Dry matter intake (DMI) is a function of animal and dietary factors affecting

hunger and satiety (Allen, 2000). Dry matter intake starts to decrease a few weeks before

parturition with the lowest level occurring at calving (Ingvartsen and Andersen, 2000).

Average values for the prefresh transition period have been reported to range between 1.7

and 2.0% of body weight (BW) (Hayirli et al., 1999). However this is not a constant value

and it can be influenced by the ration that is fed (concentration of nutrients), the stage of

the transition period, body condition score (BCS) and parity (Hayirli et al., 2002). Dry

matter intake decreases about 32% during the final three weeks of gestation, and 89% of

that decline occurs at five to seven days before calving (Hayirli et al., 2002). Most cows

rapidly increase DMI for the first three weeks after calving (Ingvartsen and Andersen,

2000). As a percentage of body weight, heifers consume less feed than cows at 21 days

before calving. By the time of calving, intake is more similar (Table 2-1) (Hayirli et al.,

1999).

Table 2-1. Predicted changes in dry matter intake for Holstein dairy cows duringthe last 3
weeks prior to calving
Cows DMI (Kg/d) DMI, % BW % Change
D-21 D-l D-21 D-1
Heifer (605 kg) 10.3 7.4 1.71 1.22 -28
Cow (740 kg) 14.4 9.8 1.94 1.33 -32
Adapted from: Hayirli, A., R. R. Grummer, E. Nordheim, P. Crump, D. K. Beede, M. J.
VandeHaar, L. H. Kilmer,, J. K. Drackley, D. J. Carroll, G. A. Varga, and S. S. Donkin.
1999. Prediction equations for dry matter intake of transition cows fed diets that vary in
nutrient composition. J. Dairy Sci. 82(Suppl. 1):113.


The National Research Council (2001) reported that DMI as percentage of body

weight during the last 21 days of gestation for heifers is equal to 1.71 0.69 e 035t





6

and for cows is equal to 1.97 0.75 e .16t, where "t" is equal to days of pregnancy minus

280.

Glucose and Lipid Metabolism

Glucose and amino acids are the major fuel supply of the developing fetus in

ruminants. Glucose and amino acids are also needed by the mammary gland for lactose

and milk protein synthesis, respectively (Herdt, 2000). Glucose demand in Holstein cows

has been estimated at 1000 to 1100 g/d during the last 21 d of gestation, but increases

sharply after calving to approximately 2500 g/d at 21 d postpartum (Drackley et al., 2001).

Ruminants are not entirely dependent on dietary glucose; as a result they are in a constant

stage of gluconeogenesis (Herdt, 2002). The liver serves as a linchpin in adaptation to the

maintenance of body fuel supplies and consequently it is the key regulator of glucose

supply to the tissues (Herdt, 2000). The major gluconeogenic precursor in ruminants is

propionic acid produced in the rumen. Its contribution to gluconeogenesis has been

estimated to be 32 to 73% (Seal and Reynolds, 1993). Liver uptake ofpropionate by portal

circulation is almost 100% (Herdt, 2002); however the capacity of the liver to convert

propionate to glucose seems to be responsive to the amount ofpropionate supplied and the

physiological stage of the animal (Drackley et al., 2001). Hepatic propionate metabolism

is modulated during the transition period. As an example, hepatic blood flow in cows

increases 84% from 11 d prepartum to 11 d postpartum (Reynolds et al., 2000). In

addition, propionate conversion to glucose by the liver is 19 and 29% greater at day 1 and

21 postpartum, respectively, than at day 21 prepartum (Overton et al., 1998).

Amino acids, lactate and glycerol are secondary substrates for gluconeogenesis in

ruminants (Herdt, 2002). Contribution to glucose production has been estimated to be 10

to 30% for amino acids, 15% for lactate and small amounts for glycerol (Seal and





7

Reynolds, 1993). Similar to propionate, the contribution of these secondary glucose

precursors is partially dependent upon their supply and metabolic adaptation of transition

dairy cows (Drackley et al., 2001). Skeletal muscle, and to a lesser extent, skin, through

suppression of tissue protein synthesis and possibly increased proteolysis, serves as a

labile pool of amino acids that is mobilized to support increased gluconeogenesis during

the transition period (Bell, 1995; Bell et al., 2000). Alanine and glutamine account for 40

to 60% of the glucogenic potential of all the amino acids; therefore they typically make

the greatest contribution to glucose synthesis (Drackley et al., 2001). Alanine conversion

to glucose at 1 and 21 days postpartum was 198 and 150%, respectively, of that at 21 day

prepartum (Overton et al., 1998). Lactate utilization for gluconeogenesis primarily

represents recycling of carbons because most circulating lactate is formed either during

catabolism of glucose by peripheral tissues or by partial catabolism of propionate by

visceral peripheral tissues (Drackley et al., 2001). However, when non-forage fiber

sources and monensin were fed to transition prepartum cows, pyruvate carboxylase

expression was significantly induced at calving, suggesting an increased capacity of

peripartum cows for gluconeogenesis from lactate (Williams et al., 2003).

Non-esterified fatty acids (NEFA) concentrations are maximum at parturition

(0.9 to 1.2 mEq/L) with a slow decrease after 3 days postpartum (Melendez et al., 2002).

This finding corroborates the elevated fat mobilization occurring around parturition in

dairy cattle. Extreme rates of lipid mobilization lead to increased uptake of NEFA by liver

and increased triglyceride (TG) accumulation (Drackley, 1999). When blood glucose

concentrations increase lipogenesis predominates over lypolysis. This results in

suppression of NEFA release from adipose tissue (Herdt, 2000). The effect of glucose on

adipose tissue is related to insulin secretion and its role in glycerol synthesis, which is





8

essential for triglyceride assembly (Herdt, 2000). When glucose concentration decreases,

as occurs just after calving, NEFA mobilization from adipose tissue is stimulated (Herdt,

2000; Melendez et al., 2002). When blood glucose levels are sufficient, glucose flow is

favored into the Krebs cycle, therefore its precursors are slowly accumulated in

mitochondria. In liver, the excess of citrate is mobilized out of the mitochondria and

converted to malonyl CoA. This intermediate inhibits the enzyme carnitine acyltransferase

I (CAT I), which inhibits the oxidation of fatty acids and indirectly stimulates TG

synthesis. Contrary, when glucose is low, malonyl CoA decreases, CAT I is activated,

favoring the transport of NEFA into the mitochondria, with the consequent increase in

ketogenesis (Nelson and Cox, 2000).

Endocrine regulation of gluconeogenesis, ketogenesis and lipid metabolism

includes insulin, glucagon, somatotropin, catecholamines, cortisol, thyroid hormones and

leptin (Herdt, 2000, Drackley et al., 2001). Glucose levels in a prepartum dairy cow are

high until parturition (Grum et al., 1996). As a result, insulin concentrations are higher

before calving than after calving and glucagon experiences the opposite pattern (Herdt,

2000; Nelson and Cox, 2000; Drackley et al., 2001). Somatotropin is lower before calving

than post calving with a peak at parturition (Grum et al., 1996). Table 2-2 summarizes the

effect of different hormones on carbohydrate and lipid intermediary metabolism in dairy

cattle.


Calving-Related Disorders

The majority of diseases that affect dairy cows occurs during the peripartum

period, consequently they are also called calving-related disorders (Risco and Melendez,

2002). In general, health disorders present low heritabilities (h2= 0 0.05) and

environmental management plays the most important role in decreasing or preventing their





9

incidence. Some exceptions are lameness (h2=0.16) and ketosis (h2= 0.39) and the

selection on conformation traits can help reduce the incidence of disease, although genetic

correlations are low (Van Dorp et al., 1998).

There are several studies that have described the relationship and risk factors

among calving-related disorders in dairy cattle (Table 2-3) (Curtis et al., 1984; Curtis et

al., 1985; Erb et al., 1985; Grohn et al., 1989; Erb and Gr6hn, 1988; Correa et al., 1990;

Correa et al., 1993; Bruun et al., 2002; Melendez et al., 2003a; Melendez et al., 2003b).

Results in general have been consistent, but some weaknesses of these studies have been

the variability of the case definition. In an effort to homogenize criteria definitions, Kelton

et al. (1998) recommended some guidelines for recording and calculating selected clinical

diseases in dairy cattle (Table 2-4).

Calving-related disorders result in significant economic losses to dairy producers

through reduction in reproductive performance and milk yield during the subsequent

lactation, cost of treatments and increased culling (Curtis et al., 1984; Curtis et al., 1985;

Erb et al., 1985; Grohn et al., 1989; Erb and Gr6hn, 1988; Correa et al., 1990; Correa et

al., 1993; Bruun et al., 2002; Risco and Melendez, 2002).

The most relevant calving-related disorders are milk fever or hypocalcemia,

retained fetal membranes, metritis, ketosis, displacement of the abomasum, mastitis and

lameness (Correa et al., 1993; Goff and Horst, 1997b; Risco and Melendez, 2002).

In Table 2-4 case definitions, economic losses, epidemiology and recommended

guidelines for recording and calculating selected calving-related disorders in dairy cattle

are reported.





10

Table 2-2. Effect of hormones on carbohydrates and lipids intermediary metabolism on
dairy cattle.
Hormone Effect on carbohydrates Effect on lipids

Insulin T glucose transport into cells 4, lipolysis
4- gluconeogenesis T lipogenesis
f glycogen synthesis
4- glycogenolysis
t glycolisis

Glucagon T gluconeogenesis T lipolysis
t glycogenolysis I ketogenesis ?
T glucose export
4 glycolisis
-1 glycogen synthesis

Catecholamines 1 glycogenolysis T lipolysis
T gluconeogenesis
T glucagons secretion
4 insulin secretion

Growth t blood glucose 4- lipogenesis
Hormone T NEFA mobilization

Cortisol 1 gluconeogenesis from proteins T lipolysis

Adapted from:
Herdt, T. H. 2000. Ruminant adaptation to negative energy balance. Influences on the
etiology of ketosis and fatty liver. Vet. Clin. North Am. Food Anim. Pract. 16:215-230.
Nelson, D. L. and M. M. Cox. 2000. Oxidation of fatty acids. Pages 598-622. In
Lehninger Principles of biochemistry. Third edition. Worth Publishers, New York, NY
10010.
Drackley, J. K., T. R. Overton, and G. N. Douglas. 2001. Adaptations of glucose and long-
chain fatty acid metabolism in liver of dairy cows during the periparturient period. J.
Dairy Sci. 84(E. Suppl.):E100-E112.
Herdt, T. H. 2002. Gastrointestinal physiology and metabolism. Postabsorptive nutrient
utilization. Pages 303-322 in Textbook of Veterinary Physiology. Third Edition. J.
Cunningham. W.B. Saunders Company





11


Table 2-3 Summary of relationships among calving-related disorders
Author Disease Risk factors Association
Curtis et Milk fever Parity Positive
al., 1984 Estimated transmited Positive
ability
Increased ccrude protein Negative
in dry period

Erb et al., RFM Milk fever OR= 2.0
1985 Parity Positive

Milk fever Parity Positive

Metritis Milk fever OR= 1.6
RFM OR= 5.8

Curtis et Milk fever Parity Positive
al., 1985
RFM Milk fever OR= 4.0
Parity Positive

Metritis RFM OR= 5.7
LDA OR= 3.6

LDA Ketosis OR= 11.9

Ketosis LDA OR= 53.5
RFM OR= 16.4
Milk fever OR= 23.6

Gr6hn et Milk fever Parity Positive
al., 1989 Milk yield Positive

Udder edema Parity Negative
Milk yield Positive
RFM OR= 2.6
Mastitis OR= 3.8

Milk Fever OR= 2.5
Abomasum Ketosis OR= 5.7
disorders Hypomagnesemia OR= 7.0
RFM OR= 2.4
Metritis OR= 2.5
Mastitis OR= 3.6





12

Table 2-3 Continued
Author Disease Risk factors Association

Gr6hn et Ketosis Parity Positive
al., 1989 Milk yield Positive
continued Milk fever OR= 1.6
LDA OR= 2.5
Metritis OR= 2.3
Mastitis OR= 1.4
Lameness OR= 2.4

Correa et Milk fever Lead feeding OR= 2.4
al., 1990
RFM Low Ca diets OR= 1.7
Farmer treatments OR= 1.7

Metritis Dystocia OR= 5.6
RFM OR= 86.5

LDA Metritis OR= 43.7
Leading feeding OR= 4.4

Ketosis Milk fever OR= 41.5
Dry fat cows OR= 3.1

Mastitis RFM OR= 8.7
Milk fever OR= 31.3

Correa et RFM Dystocia OR= 2.2
al., 1993 Twinning OR= 3.4

Metritis Dystocia OR= 2.1
RFM OR= 6.0
Ketosis OR= 1.7


Ketosis Milk fever OR= 2.4


LDA Milk fever OR= 2.3
Ketosis OR= 13.8
Dystocia OR= 2.3


Dystocia Twinning OR= 10.5
Milk fever OR= 2.6





13

Table 2-3 Continued
Author Disease Risk factors Association
Collard et Laminitis Energy balance postpartum Negative
al., 2000
Digestive Energy balance postpartum Negative

Bruun et Metritis Dystocia Positive
al., 2002 Reproductive disease Positive
Retained placenta Positive

Schnier et Mastits Warm housing system Positive
al., 2002
Metritis Cold housing system Negative

Melendez Ovarian cysts Lameness Positive
et al.,
2003a

Melendez Ketosis Displacement of abomasums Positive
et al.,
2003b Retained fetal membranes Positive
Displacement of
abomasums Ketosis Positive
Parity Positive
Metritis Retained fetal membranes Positive
Parity Negative





14

Table 2-4. Case definition, incidence and economic losses ofcalving-related disorders
Disease Case definition Incidence Economic losses

Milk fever Calcium deficiency causing Median 6.5% $335 per case
progressive neuromuscular Range .03 -
dysfunction with flaccid paralysis, 22.3%
circulatory collapse, and depression of
consciousness
RFM Fetal membranes visible at the vulva Median 8.6% $285 per case
or in vagina or uterus by vaginal Range 1.3 -
examination more than 24 h after 39.2%
parturition
Metritis Abnormal cervical discharge, vaginal Median 10.1% Treatment,
discharge, or both or uterine content. Range 2.2 increased days
New case if cow did not have a case 37.3% open and culling
during the preceding 30 days
Ketosis Primary: Decreased appetite, elevated Median 4.8% $145 per case
milk, urine or breath ketones in the Range 1.3 -
absence of other disease 18.3%
LDA Decreased appetite accompanied by an Median 1.7% $340 per case.
audible, high pitched tympanic Range 0.3 Milk losses 250-
resonance (ping) by percussion of the 6.3% 2000 kg/
left abdominal wall between the 9th lactation
and 12th ribs
Ovarian Smooth, rounded structure greater Median 8.0% $39 per case
cysts than 25 mm in diameter in one or both Range 1.0 -
ovaries non pregnant cows 16.0%
Lameness Episode of abnormal gait attributable Median 7.0% $302 per case
to either the foot or leg regardless of Rangel.8 -30%
etiology or duration
Mastitis Visually abnormal milk secretion from Median 14.2%
one or more quarters with or without Range 1.7 -
signs of inflammation of the udder. 54.6%
New case following 8 days of normal
milk
Adapted from: Kelton, D. F., K. D. Lissemore, and R. E. Martin. 1998. Recommendations
for recording and calculating the incidence of selected clinical diseases of dairy cattle. J.
Dairy Sci. 81:2502-2509.





15

Milk Fever, Parturient Paresis, Hypocalcemia

Milk fever (MF), is a non-febrile metabolic disease affecting milking cows in

which acute calcium deficiency causes progressive neuromuscular dysfunction with

flaccid paralysis, circulatory collapse, and depression of consciousness (Oetzel and Goff,

1999).

The reported frequency of MF, based on 33 citations from 1979 to 1995 ranged

from 0.03% to 22.3%. The median lactational incidence was 6.5% (Kelton et al., 1998).

However, Oetzel and Goff (1999) established that annual incidence rate within herds may

vary from 2% to 60%. The incidence of MF reported in a Florida commercial dairy herd

using anionic diets during the prepartum period was 0.42% (2/477) (Melendez et al.,

2003b). Approximately 75% of all cases of MF occur within 24 hours of calving. An

additional 12% occur 24 to 48 hours after calving. Some cases (about 6%) occur at the

time of delivery and cause dystocia because hypocalcemia induce uterine atony (Oetzel

and Goff, 1999). Only 3% of cases occur prepartum, and only 4% occur more than 48

hours postpartum (Oetzel, 1988).

Hypocalcemia may be clinical or subclinical. Clinical signs of MF are not seen

until calcium is about 4 mg/100 ml (Goff and Horst, 1997b). Subclinical hypocalcemia

affects about 50% of all adult dairy cattle. In this case, plasma Ca concentration of

periparturient cows remained < 7.5 mg/dL, even up to 10 days after calving (Goff et al.,

1996). This condition may lead to decreased dry matter intake after calving, increased risk

of secondary diseases, decreased milk production and decreased fertility (Goff and Horst,

1997b).

Economic losses from MF include prophylactic or clinical treatment, lost milk

production, poor reproductive performance, and culling, which are estimated to total





16

approximately $335 per case (Dohoo and Martin, 1984; Erb et al., 1985; Bigras-Poulin et

al., 1990; Guard, 1994; Gr6hn et al., 1998; Kelton et al., 1998).

Several risk factors related to MF have been identified. Breed, age and milk yield

are the most important risk factors for MF in dairy cattle. Jersey and Guernsey cows are

the most susceptible to MF; Holstein and Brown Swiss are moderately susceptible; and

Ayrshire and Milking Shorthorns are the least susceptible (Oetzel and Goff, 1999). The

incidence of MF increases with parity and with higher levels of milk production,

regardless of breed (Oetzel and Goff, 1999).

Most cows are in negative Ca balance during the early weeks of lactation because

more Ca leaves the body via milk, endogenous fecal loss, and urine than is absorbed from

the diet. This is because the intestinal mechanisms for absorbing calcium are not fully

adapted to lactation and also because dry matter intake is less than favorable (Oetzel and

Goff, 1999). Bone Ca mobilization is stimulated by a concerted effort of PTH and

1,25(OH)2D, but intestinal Ca absorption is controlled by 1,25(OH)2D alone. During the

dry period, these mechanisms for replenishing plasma Ca are relatively inactive. Thus,

nearly all cows experience some degree ofhypocalcemia during the first days after calving

as the intestine and bone adapt to lactation (Goff et al., 1996). The adaptation process

begins with dramatic increases in the plasma concentrations of PTH and 1,25(OH)2D at

the onset of hypocalcemia. About 24 hours of 1,25(OH)2D stimulation is required before

intestinal Ca transport is increased significantly. Bone Ca resorption (recruitment and

activation of osteoclasts) is not significantly increased until after about 48 hours of PTH

stimulation. In cows with MF, these adaptive processes can be even more prolonged

(Horst et al., 1994). Magnesium status is another factor influencing the risk of

hypocalcemia. Low blood magnesium levels can reduce PTH secretion from the





17

parathyroid glands and can also alter the responsiveness of tissues to PTH. High dietary

potassium reduces ruminal magnesium absorption (Oetzel and Goff, 1999). Increase

dietary P intake (> 80 mg/day) increases the P in blood (- 8 mg/dL), which has a direct

inhibitory effect on the renal enzymes that catalyze production of 1,25(OH)2D. This

reduced production of 1,25(OH)2D further reduces intestinal Ca absorption mechanism

prepartum (Horst et al., 1994; Oetzel and Goff, 1999).

One of the most important determinants of MF risk is the acid-base status of the

animal at the time of parturition. Metabolic alkalosis appears to alter the physiologic

activity of PTH so that bone resorption and production of 1,25(OH)2D are impaired, thus

reducing the ability of the animal to successfully adjust to increased calcium demands

(Oetzel and Goff, 1999).

Milk fever is confirmed by low serum calcium concentrations. Clinical signs may

begin as total blood calcium values fall below 7.5 mg/dL (< 1.8 mmol/L). However more

than half of all mature dairy cows will have total blood calcium concentrations below 7.5

mg/dL (< 1.8 mmol/L) after calving without any evidence of clinical signs (Goff, 1999a;

Oetzel and Goff, 1999). Generally, cows that are recumbent and unable to rise as a result

of low blood Ca will have plasma Ca concentration less than 5 mg/dL, and some will be

down as low as 2 mg/dL; below this level is generally incompatible with life (Goff,

1999a). Milk fever presents three stages of clinical symptoms. Animals in stage I usually

have levels of 5.5 to 7.5 mg/dL of Ca. Animals in stage II typically have a total Ca

concentration of 3.5 to 6.5 mg/dL. Animals in stage III may have Ca levels as low as 1.0

mg/dL. Blood concentrations of P are typically below normal and magnesium

concentrations are usually high. The most important differential diagnosis is related to the

downer cow syndrome (Oetzel and Goff, 1999).





18

Stage I may be treated with either oral calcium supplements or intravenous

calcium salts. Animals in stage II or II require immediate treatment with intravenous

calcium salts (Oetzel and Goff, 1999). The fastest way to restore normal plasma Ca

concentration is to administer an intravenous injection of Ca salts (Goff, 1999a). Calcium

gluconate or borogluconate is the standard intravenous treatment in cows. Five hundred ml

of these products provide 10.8 g of calcium. Commercial preparations usually supply

between 8.5 to 11.5 g Ca/500 ml. They may also contain Mg, phosphite and glucose. All

the preparations effectively raise total and ionized Ca concentrations in blood (Goff,

1999a; Oetzel and Goff, 1999). Intravenous Ca should always be administered slowly to

prevent sudden cardiac arrest due to hypercalcemia (Oetzel and Goff, 1999). Calcium

should be administered at a rate of 1 g/minute (Goff, 1999a). Approximately 60% of

recumbent animals affected with uncomplicated MF will get up within 30 minutes after a

single intravenous dose with calcium salts. Another 15% can be expected to rise within the

next 2 hours. Full restoration of normal calcium homeostasis usually requires 2 or 3 days.

About 10% of dairy cows with MF stay recumbent for over 24 hours but eventually

recover (Oetzel and Goff, 1999).

Ca salts can also be injected subcutaneously. The serum Ca concentration achieved

is not the same as with intravenous administration with similar dose. However, as with

intravenous Ca injection, the subcutaneous dose of Ca increases Ca in blood for 4 to 5

hours only (Goff, 1999a). Greater amount of Ca can cause local tissue necrosis; then Ca

injections should be limited to 1 to 1.5 g Ca (50 to 75 mL of most commercial

preparations) per site. Calcium chloride solutions are not well tolerated subcutaneously.

Ca solutions containing glucose may also be slightly more injurious. Preparations are

available for intramuscular administration of Ca in the form of calcium levulinate or





19

calcium lactate, which tend to be less injurious to tissues than other forms of calcium

(Goff, 1999a).

A variety of oral calcium salt preparations are available for cattle (Oetzel and Goff,

1999). Oral Ca supplement must be readily soluble in water (digestive fluids) to reach the

minimum amount for passive transport (- 6 mmol/L) (Goff, 1999a). About 4 g of calcium

will be absorbed and enter the bloodstream of a cow given an oral solution containing 50 g

of calcium chloride (Oetzel and Goff, 1999). Calcium chloride is the most soluble of the

Ca salts; calcium propionate, calcium formate, calcium acetate, calcium gluconate and

calcium lactate are also soluble enough. Calcium hydroxide, calcium oxide and calcium

carbonate are relatively insoluble and unsuitable for treating hypocalcemia. Ca that is not

absorbed by passive diffusion is still available for absorption by active Ca transport in the

small intestine, but this absorption is not rapid enough to be of aid in the treatment of

hypocalcemia (Goff, 1999a). Oral Ca products typically contain between 25 and 100 g of

Ca (Oetzel and Goff, 1999). Calcium chloride and calcium propionate are the most

common products used in the treatment and prevention of hypocalcemia in cattle (Goff,

1999a). Oral administration of 50 g of calcium from calcium chloride as a drench in 250

mL of water raises plasma Ca concentrations to the same extent as 4 g of Ca as calcium

chloride administered intravenously. Conversely, 100 g of Ca orally is equivalent to

between 8 and 10 g of calcium administered intravenously. Calcium chloride increases

plasma Ca better than calcium propionate (Goff and Horst, 1993). Calcium chloride is

slightly more effective and takes up less volume than calcium propionate; it also is an

acidifying agent. This mild metabolic acidosis can help enhance Ca homeostasis.

However, severe metabolic acidosis can be produced when repeated treatments of calcium

chloride are administered. Furthermore, calcium chloride is very irritating to mucous





20

membranes and lesions can be produced in the upper digestive tract, rumen and

abomasum (Goff 1999a). Calcium absorption in the rumen is less efficient than in the

intestine because the volume of fluid in rumen will rapidly dilute the Ca concentration to a

value less than 6 mmol/L required for passive absorption. Pastes and gels reduce the

amount of Ca likely to bypass the rumen. Thus, in general, more Ca must be administered

by gel or paste to obtain the same rise in plasma Ca as is achieved by an oral Ca drench.

However, drenches present the disadvantage that some cows will aspirate the solution,

which can lead to severe aspiration pneumonia (Goff, 1999a). Calcium propionate is

effective and less irritating to tissues than is calcium chloride. It does not induce metabolic

acidosis, so larger amounts of Ca can be given. Furthermore, it supplies the cow with a

gluconeogenic precursor (propionate) (Goff, 1999a). Oral treatments increase Ca within

30 to 60 minutes of administration, and plasma Ca concentrations remain elevated for

about 6 hours. Calcium chloride acts a little faster, but calcium propionate may act a little

longer (Goff, 1999a).

For many years, the traditional method of preventing MF in dairy cows was the

restriction of dietary intake of Ca during the prepartum period. Ca diets with < 15-20 g of

Ca/d fed during the last 10 days of gestation, followed by a postpartum diet that is high in

Ca have been recommended. These diets will greatly reduce the risk of MF (Horst et al.,

1994; Joyce et al., 1997; Vagnoni and Oetzel, 1998 ; Oetzel and Goff, 1999). According to

NRC (2001), the requirements of Ca for a 680 kg mature dry pregnant cow (maintenance

plus last 2 month of gestation) is 0.45% dry matter basis.

Oral and intramuscular doses of vitamin D3 have also prevented MF successfully.

However, repeated treatments may lead to toxicity (Jorgensen, 1974; Markusfeld, 1989).

Parathyriod hormone has been also reported to prevent parturient paresis in dairy cows.





21

Parathyroid hormone administration increases plasma concentration of 1,25 dihydroxy

vitamin D and hydroxyproline prior to parturition, suggesting that both intestinal Ca

absorption and bone Ca resorption are increased by administration of the hormone (Goff et

al., 1989).

Dietary cation-anion difference has been defined as the difference in

milliequivalents of cations and anions per kilogram of dry matter and has a direct impact

on blood acid-base metabolism (Block, 1994). Important dietary cations are sodium (Na),

potassium (K), calcium (Ca) and magnesium (Mg); important dietary anions are chloride

(Cl), sulfur (S) and phosphorus (P). Several methods of calculating DCAD have been

utilized, including the following equations:

DCAD (mEq)= (Na + K)-(Cl + S) (3-1)

DCAD (mEq)= (Na + K) (Cl) (3-2)

DCAD (mEq)= (Na + K + 0.15 Ca + 0.15 Mg)-( Cl + 0.2 S + 0.3 P) (3-3)

DCAD (mEq)= (Na + K + 0.15 Ca + 0.15 Mg) (Cl + 0.25 S + 0.5 P) (3-4)

Equations 3-1 and 3-2 are used more commonly, but it must be kept in mind that

Ca, Mg, and P absorbed from the diet will also influence blood pH. Equations 3-3 and 3-4

take into account new data on the bioavailability of all the potential strong ions.

Theoretically it should be the most accurate, but it is has not been widely applied (Goff

and Horst, 1998a; Oetzel and Goff, 1999).

In lactating dairy cows, increasing dietary cation-anion difference increased DMI

and milk production in early and midlactation. These effects were not observed in late

lactation. However, increased DCAD affected acid-base parameters in urine at all stages

of lactation (Delaquis and Block, 1995).





22

Diets fed prior to parturition that evoke an acidic response in the animal reduce

MF risk, whereas diets that evoke an alkaline response increase it. Thus, low DCAD diets

cause metabolic acidosis and reduce the risk of MF. A diet can have a low DCAD because

it is low in cations, high in anions, or a combination of both (Oetzel and Goff, 1999). As

DCAD decreases, H+ increases, HCO'3 decreases and pH decreases. These changes are

accompanied by a reduction in urinary HCO'3 excretion and urinary pH as compensatory

mechanisms. Furthermore, low DCAD prepartum can mitigate hypocalcemia via increased

urinary Ca reabsorption, serum ionized Ca, and responsiveness to Ca homeostatic

hormones (Block, 1994; Vagnoni and Oetzel, 1998). Typical diets fed to dry cows have a

DCAD of about +50 to +250 mEq/kg of diet dry matter (using equation 1). In common

feedstuffs, potassium is the most variable of the ions in the DCAD equation and it is

usually the most important determinant of DCAD in non-supplemented feed (Oetzel and

Goff, 1999). The successful use of dietary anions to prevent MF has suggested that diets

that are high in cations, especially Na and K, increase the susceptibility of cows to MF. A

good first step in formulating a low DCAD prepartum diet is to reduce dietary potassium

to less than 1.5% of dry matter. Once the cation content has been reduced as much as

possible by diet selection, anions can then be added to further reduce DCAD to the desired

end point (Oetzel and Goff, 1999). Commonly used anion sources are calcium chloride,

ammonium chloride, magnesium sulfate, ammonium sulfate, and calcium sulfate. Anionic

salts can be unpalatable and are always accompanied by a cation, which, depending on its

rate of absorption, will counteract some of the effects of the anions (Goff and Horst,

1998a, 1998b). Other anion sources include mineral acids such as hydrochloric or sulfuric

acid (Oetzel and Goff, 1999). However these acids in a liquid form are corrosive and

dangerous to handle. Commercial preparations of HCI mixed into common feed





23

ingredients as a premix could offer an inexpensive, safe and palatable alternative to

anionic salts (Goff and Horst, 1998b). Optimal acidification generally occurs when anions

are added to achieve a final DCAD (using equation 1) between -50 to -150 mEq/kg of dry

matter. The strong negative relationship (r2=0.95) between urinary pH and net acid

excretion by cows fed the diets containing anionic salts suggests that urinary pH

measurement might be a useful tool to assess the degree of metabolic acidosis that is

imposed by dietary anionic salts (Vagnoni and Oetzel, 1998). An advantage of this

approach is that it accounts for inaccuracies in mineral analyses and for unexpected

changes in forage mineral content. Urinary pH can be evaluated by obtaining urine from a

group representing about 10% of the pre-calving cows. Urinary pH values below 5.5

indicate overacidification and DCAD should be increased. The optimal urinary pH is

between 6.0 and 6.5 for Holstein cows and between 5.8 and 6.2 for Jersey cows. Over 6.5

is considered inadequate acidification and suggests that a lower DCAD is required. In

herds experiencing MF the urine of close-up dry cows will be very alkaline with a pH

above 8.0. Most accurate results will be obtained by collecting urine samples at a standard

time, preferably within a few hours after feeding (Goff and Horst, 1998a; Oetzel and Goff,

1999).

Retained Fetal Membranes Metritis Complex

Retained Fetal Membranes (RFM) is defined as the lack of detachment of fetal

membranes from the maternal caruncles within the first 12 to 24 hours after calving

(Grunert, 1986; Eiler, 1997). Van Werven et al., (1992) reported that 77.3% of cows

expel the fetal membranes by 6 hours. Normal expulsion of fetal membranes requires that

the maternal and fetal tissues undergo maturation and a loosening process, which is

completed by 2 to 5 days before the end of an average gestation. These changes include





24

collagenization of connective tissue, reduction in blood supply, appearance of polynuclear

giant cells, loosening of tissues, and contraction of the uterine musculature (Grunert, 1986;

Youngquist and Braun, 1993). Fifty-nine percent of RFM were spontaneously expelled

between 5 and 7 days postpartum and 94% at days 11-13 postpartum (Van Werven et al.,

1992). Most RFM are expelled by 4 to 10 days postpartum after sufficient necrosis of the

caruncular tissue (Youngquist and Braun, 1993). Significant changes in the placentome

must take place before expulsion of the fetal membranes can occur. Increased collagenase

and other protease activities have been described in the normal uterus. This has resulted in

a massive breakdown of collagen and other proteins during uterine involution (Sharpe et

al., 1990). However, in cows experiencing RFM collagenolysis and proteolysis is

diminished (Eiler, 1997).

Some studies have reported an average incidence of RFM from 4 to 11% with a

range of 2-55% (Paisley et al., 1986; Joosten et al., 1988). The incidence of RFM, based

on 50 citations from 1979 to 1995 ranged from 1.3% to 39.2%. The median was 8.6%

(Kelton et al., 1998). Economic losses of a case of RFM, including direct cost such as

treatment (32%), lost milk production (40%), increased culling rate (19%) and increased

days open (9%), have ranged from $106 to $285 (Barlett et al., 1986a;Joosten et al., 1988;

Guard, 1994).

Joosten et al., (1991) have reported a number of risk factors for RFM such as

dystocia, parity, abnormal gestation duration, season and sire of the calf. In other studies,

cows that developed parturient paresis were 4.0 to 4.2 times more likely to experience

RFM than normal cows (Curtis et al., 1985; Erb et al., 1985).

Hypocalcemia has been related to dystocia, RFM and uterine prolapse in dairy

cattle (Curtis et al., 1983; Risco et al., 1984; Correa et al., 1993; Risco et al., 1994a). Erb





25

et al., (1985) determined that cows suffering hypocalcemia were 4.2 times and 2.0 times

more likely to have dystocia and RFM, respectively.

A recently explored, but old concept related to RFM is the immunological aspect

involved in the pathogenesis of this reproductive condition. The old theory suggests that

fetal membranes must be recognized as "foreign" tissues and rejected by the immune

system after parturition to cause expulsion of fetal membranes (Gunnink, 1984).

Neutrophils isolated from cows that experienced RFM had significantly lower function

than cows without RFM, before calving and during the first 2 weeks postpartum. In

addition, Interleukin-8, a potent neutrophil chemoattractant, was lower at calving in cows

with RFM (5112 pg/ml) than cows without RFM (13411 pg/ml). These findings suggest

that neutrophil function around parturition is a determining factor for the development of

RFM in dairy cattle (Kimura et al., 2002).

Retained fetal membranes have been the major factor that predispose cattle to

metritis. The majority of affected cows show no serious clinical signs other than a

transient decrease in appetite and milk production. However, 20% to 25% of cows

affected by RFM develop moderate to severe metritis (Joosten et al., 1988).

The relative risk of cows with RFM developing metritis was 2.8 and the

attributable risk was 28% as compared with cows without RFM (Bartlett et al., 1986). In

several epidemiological studies using path analysis methodology cows with RFM have

been around 6.0 times more likely to develop metritis than normal cows (Correa at al.,

1993; Curtis et al., 1985; Erb et al., 1985). Overall, studies using path analysis and risk

assessment indicated consistently that dystocia, nutrition, metabolic disorders and mainly

RFM increased the likelihood that a cow would develop metritis (Lewis, 1997; Bruun et

al., 2002; Melendez et al., 2003b).





26

Uterine infections are one of the most frequent disorders affecting dairy cows

during the post partum period (Youngquist and Shore, 1997). They are a major cause of

economic losses to the cattle industry being related with systemic disease, decrease

reproductive efficiency, reduced milk production, increased replacement costs and

reduced genetic progress (Kelton et al., 1998). The total cost to producers for each

lactating dairy cow with a uterine infection was estimated at $106 (Barlett et al., 1986a).

In a review by Fourichon et al., (1999), only two studies out often showed losses in milk

production between 100 to 270 kg per lactation in cows with metritis. Typically, no milk

loss was associated with metritis. Rajala-Schultz and Grihn (1998) using data from

37,776 Finish Ayrshire dairy cows determined that when metritis was treated as one

disease complex, regardless of the time of its occurrence, metritis had no significant effect

on milk yield. However, when early and late metritis were analyzed separately, the time of

disease occurrence had an effect on milk yield. Late metritis was not associated with milk

loss yet early metritis accounted for 46.2 kg less milk less in cows with metritis within 28

days after calving.

Barlett et al. (1986a) in Michigan herds determined that cows with metritis were

1.3 times more likely to be culled than were cows without metritis. A more recent study in

Holstein cows in New York State demonstrated that metritis had no effect on the risk of

culling. In contrast, other diseases such as mastitis, MF, RFM, LDA, ketosis and ovarian

cysts significantly affected culling at different stages of lactation (Gr6hn et al., 1998).

The diagnosis of metritis has been very subjective and has been applied to clinical

conditions that range from cows that are nearly normal to those affected by severe, life

threatening sepsis (Youngquist and Shore, 1997). Metritis has been defined as an

inflammation of all layers of the uterus typically developed within a few days to several





27

weeks after calving. The condition is characterized by an abnormal cervical discharge,

vaginal discharge, or both (Lewis, 1997; Youngquist and Shore, 1997; Kelton et al.,

1998). The condition may be local or systemic. A condition called toxic or gangrenous

metritis, that occurs almost exclusively in the puerperal period and is often associated with

clostridial infections, is characterized by foul-smelling, watery uterine discharge, severe

drop in milk production and systemic symptomatology (Gilbert and Schwark, 1992; Olson

et al., 1986; Smith et al., 1998).

Several microbiological risk factors have been associated with uterine infections.

The uterus has an anaerobic environment. Cows with RFM present a higher incidence of

coliforms, and other environmental bacteria, clostridia, Archenobacter pyogenes, and

gram-negative anaerobes than normal cows (Olson et al., 1986). Recent studies have

confirmed that A. pyogenes, either alone or in combination with anaerobic bacteria

(Fusobacterium necrophorum and Bacteroides spp.) can act to induce uterine infections in

cows during the puerperal or luteal phase (Del Vecchio et al., 1992). These bacteria

produce postpuerperal metritis or a more delayed typical endometritis (Olson et al., 1986;

Lewis, 1997; Youngquist and Shore, 1997). Postpuerperal metritis eventually become a

chronic problem. Affected cattle do not present a systemic disease but they may have a

mucopurulent, fetid vulvar discharge.

The incidence of uterine infections varies considerably among studies, and the

average incidence is not an especially meaningful statistics (Lewis, 1997). This variation

is due to poorly described diagnostic methods and lack of good definition of a case. Based

on 43 citations from 1979 to 1995, the frequency of metritis ranged from 2.2% to 37.3%.

The median was 10.1% (Kelton et al., 1998). Gr6hn et al., (1998) determined an incidence

of metritis of 4.2% in 7523 dairy cows in New York state.





28

Uterine infections can reduce the reproductive efficiency of dairy cows thereby

increasing herd health cost (Lewis, 1997). Metritis increased days from calving to first

estrus by 6.9 days, 7.3 days calving to first service interval, 15.4 days first to last service

interval, 0.3 services per conception and 18 days calving to conception interval (Barlett et

al., 1986a). Uterine infections alter uterine involution and reduce ovarian follicular

development during the early postpartum period, which may prolong the interval from

calving to estrus and AI, but the mechanism and the repeatability of the effect of the

condition on ovarian follicles is unclear (Del Vecchio et al., 1994; Lewis, 1997).

Uterine infections are a highly complex process. The exact causes of uterine

infections are unknown, but several predisposing factors have been associated with the

disease (Lewis, 1997). Cows with dystocia, RFM, twins or still-births, overconditioning

and various metabolic and digestive disorders increase the risk of metritis (Curtis et al.,

1985; Erb et al., 1985; Olson et al., 1986; Correa et al., 1993; Lewis, 1997; Youngquist

and Shore; 1997; Bruun et al., 2002; Melendez et al., 2003b). Other risk factor that might

be associated with the pathogenesis of metritis is the immunosupression occurring around

parturition. T-cell sub-populations have been demonstrated to decline at calving and they

do not return to pre-calving levels until two weeks after parturition (Kimura et al., 1999).

Treatment of RFM has been based on different protocols. Removal by gentle

traction has long been the conventional method, but the procedure may be followed by

severe uterine infections and impaired fertility (Bolinder et al., 1988). A single dose of

oxytocin does not reduce the presence of RFM (Miller and Lodge, 1984). Exogenous

estrogen is of questionable therapeutic value because plasma concentrations of estrogen

are elevated in cows when the placenta is retained (Pimental et al., 1987). Analogs of

Prostaglandin F2 alpha are recommended as well, but Fenprostalene does not change





29

myometrial activity between days 1 and 4 after calving, suggesting that uterotonic agents

are unlikely to promote placental expulsion. In addition, the levels of PGFM are higher in

cows with RFM than control cows (Burton et al., 1987; Risco et al., 1994b). Treatment of

cows with RFM with intrauterine tetracycline may reduce fertility (Youngquist and Braun,

1993). Parenteral administration of antibiotic agents is indicated in cases of sepsis

associated with RFM (Smith et al., 1998), however, the residues in milk must be

considered (Dinsmore et al., 1996).

Treatment of the uterine infections traditionally has been based on the use of local

and/or systemic antibacterial compounds. However, according to different trials, the

results have been controversial (Gilbert and Schwark, 1992; Youngquist, and Braun, 1993;

Pugh et al., 1994; Smith et al., 1998).

Uterine infections should be prevented by proper nutritional management during

the dry period, allowing cows to calve in an uncontaminated environment, and employing

strict sanitation if assistance is required during delivery (Youngquist and Braun, 1993;

Lewis 1997). Ultimately, the competence of the immune system has to be considered in

the prevention of the metritis complex. Uterine trauma, such as dystocia, manual removal

of RFM and intrauterine infusions, reduced the phagocytic activity of uterine and blood

neutrophils (Cai et al., 1994). Changes in immune function are consistent with changes in

estradiol and progesterone concentrations around calving and during the estrous cycle of

cows and ewes. Prostaglandins and other arachidonic acid metabolites might be important

mediators of resistance or susceptibility to uterine infections (Lewis 1997; Goff and Horst,

1997). Repeated doses of prostaglandin F2alpha beyond 7 days postpartum in cows with

metritis resultes in less acute response protein concentrations and lower diameter of





30

uterine horns. This response might be explained for the effect of PGF2alpha on smooth

musculature contraction of the uterus (Melendez et al., 2003c).

Abomasal Disorders

Displacements, dilatations, and volvulus of the abomasum are the most commonly

encountered disorders of the gastrointestinal tract in modem dairy operations (Trent,

1990). Displacement can be on the left side (LDA) or the right side (RDA). (Fecteau et al.,

1999). Omental attachments of the abomasum prevent true torsion around the long axis of

the abomasum, with rotation occurring around an axis through the supporting lesser

omentum. Therefore, a more accurate term for the syndrome is "abomasal volvulus",

rather than torsion. Any right-sided displacement that requires further manipulation to free

the pylorus and duodenum may be considered for practical purposes to be a volvulus

(Trent, 1990).

The prevalence of abomasal displacement among dairy herds is variable depending

on geographic location, management practices, climate and other factors (Fecteau et al.,

1999). Left displacement is the most common of the three syndromes, constituting 85 to

95.8% of the total cases of displacements or volvulus (Trent, 1990).

Left displacement of abomasum

In LDA, the abomasum slides under the rumen and dorsally along the left body

wall. The result is a partial impairment of abomasal outflow, leading to abomasal gas

accumulation, electrolyte pooling with subsequent systemic alterations, and depressed

gastrointestinal motility and appetite (Fecteau et al., 1999). A simple case definition is a

cow with decreased appetite accompanied by a progressive decrease in milk production.

An audible, high pitched tympanic resonance (ping) produced by percussion of the left

abdominal wall between the 9h and 12h ribs is a characteristically used to diagnose this





31

condition (Kelton et al., 1998). Feces are often softer than normal. Rectal temperature,

respiratory and heart rate are generally normal (Fecteau et al., 1999).

Left displacement of the abomasum occurs most commonly two weeks pre- to 8

weeks postpartum. The incidence of LDA, based on 22 citations from 1982 to 1995

ranged from a postpartum incidence rate of 0.3% to 6.3%. The median incidence rate was

1.7% (Kelton et al., 1998). Detilleux et al., (1997), in New York State, determined an

incidence of LDA that ranged from 2.1 to 8.7%. The mean number of days of lactation at

which LDA was diagnosed was 20.6. In New York state, Gr6hn et al., (1998) observed a

lactational incidence risk of LDA of 5.3%. Cameron et al., (1998) in 1170 multiparous

Holstein cows from 67 high producing dairy herds in Michigan, found an incidence of

LDA of 6% in primiparous and 7% in multiparous cows. Cows with twins presented an

incidence of 11 to 12 %. Ostergaard and Gr6hn (1999) in three Danish research herds

(4414 lactations) determined an incidence of LDA in first lactation cows of 0.6% and in

older cows 1.2%, with a first diagnosis at 19 days in milk (mean) or 17 days in milk

(median). In Florida, Massey et al, (1993) and Melendez et al. (2003b) found an LDA

incidence of 2.4% and 3.9%, respectively.

Economic losses from LDA include lost milk production and the cost of the

surgery which is estimated to total approximately $340 per case (Guard, 1994; Kelton et

al., 1998). The effect of LDA on test day milk yields of Holstein cows have been studied

by several authors. Detilleux et al., (1997) in New York State and using 12,572 Holstein

cows between parity 1 and 6, determined that from calving to 60 days after diagnosis,

cows with LDA yielded on average 557 kg less milk than did cows without LDA. Thirty

percent of losses occurred before diagnosis and milk loss increased as parity and

productivity increased. In each parity, the lactation curves of cows with LDA were





32

depressed in early lactation compared with the curves for cows without LDA. Cows with

LDA experienced severe losses of milk yield because the disease occurred during peak

yield, which might explain the severity of the losses. Milk yield returned to a normal range

at 20 to 45 d after diagnosis. Furthermore, cows with LDA were 1.8 times more likely to

have another disease than healthy cows. Ostergaard and Grohn (1999), in Denmark, found

that cows with LDA compared with healthy cows had an average milk loss within the first

6 weeks after diagnosis of 4.6 and 5.2 kg/d for primiparous and multiparous cows,

respectively. Three weeks before diagnosis, only multiparous cows with LDA showed

lower milk yield than normal cows (approximately 4.0 kg/d). Also, this study

demonstrated cows that experienced LDA presented the highest body weight loss (69 kg)

and the pre-disease level for accurate estimation of losses has to be considered.

Displaced abomasum is also a risk factor for culling in early lactation (1 to 30

days), but not in late lactation. Cows with LDA had a relative risk of culling 2.4 times

higher than normal cows during the first 30 days of lactation. After that period of lactation

the differences were not significant (Grohn et al.,1998).

Left displacement of the abomasum is a multifactorial disease where different risk

factors have been established. Several epidemiological and clinical trials have been

conducted to determine association among factors and physiopathological mechanism.

Cows experiencing MF, dystocia and ketosis increased the odds for LDA by 2.3, 2.3 and

13.8 respectively, when compared to normal cows (Correa et al., 1993). In another study

conducted by Cameron et al., (1998) in Michigan, individual cow and herd risk factors

were examined. Significant factors associated with an increased risk of LDA in the

individual cow model included high body condition score, winter season, and plasma

NEFA concentration > 0.3 meq/L between 35 and 3 d prepartum. The risk of LDA





33

decreased as lactation number increased. At the level of the herd, factors associated

positively with risk of LDA were Predicted Transmitted Ability (PTA) for milk

production, BCS, winter and summer seasons, and precalving rations containing energy

densities > 1.65 Meal of Enl/kg of DM. Feed bunk management considering bunk space,

feed availability and freshness was associated negatively with the risk of LDA.

Nutrition has been implicated as one of the most important risk factors in the

etiology of LDA. The transition period from two weeks prepartum through two to four wk

postpartum is the major risk period for the occurrence of LDA (Shaver, 1997). This might

be explained by a low feed consumption during the transition period where rumen fill is

decreased. The decline of dry matter intake is about 35% over the final week prepartum

(Bertics et al., 1992). Furthermore, as pregnancy progresses, the growing uterus occupies

an increasing amount of the abdominal cavity. This forces the abomasum forward and

slightly to the left side of the cow. After calving, if the smaller rumen does not move into

its normal position on the left ventral floor of the abdomen, the abomasum is able to slide

under it (Goff and Horst, 1997b).

In Florida, Massey et al., (1993) reported that hypocalcemic cows at parturition

(total serum calcium < 7.9 mg/dl) were 4.9 times more likely to develop LDA than

normocalcemic cows. Hypocalcemia is known to cause abomasal atony. Abomasal atony

is an absolute prerequisite to LDA (Fecteau et al., 1999). Strategies to prevent

hypocalcemia at parturition may be useful for the prevention of LDA (Shaver, 1997).

Ketosis is the strongest identified risk factor associated with LDA. Curtis et al.,

(1985) reported that cows with uncomplicated ketosis were 11.9 times more likely to

develop an LDA than normal cows. Correa et al., (1993) found that cows with ketosis

were 13.8 times more likely to develop an LDA than normal cows. Melendez et al.





34

(2003b) found that cows with clinical ketosis were 50 times more likely to develop LDA

than cows without ketosis. However this is not a cause-effect relationship. Ketosis is a

typical metabolic disease explained by low DMI, low levels of glucose, negative energy

balance and a high level of fat mobilization in early postpartum cow (Herdt and Gerloff,

1999; Herdt 2000; Risco and Melendez, 2002). Cows with low DMI at day 1 prepartum

had reduced DMI at day 21 postpartum (Grummer, 1995). Body condition score at calving

is another factor related with DMI and health status of the dairy cows. Cows with excess

BCS at parturition are at increased risk to hypocalcemia, ketosis and LDA (Cameron et al.,

1998; Heuer et al., 1999). In addition, cows with longer and more extreme period of

negative energy balance have increased risk of digestive problems, including LDA and

laminitis (Collard at al., 2000).

The review of Shaver (1997) establishes that there are no conclusive studies

reporting that increasing concentrate intake during the last two to three weeks prepartum

reduces postpartum disorders. Curtis et al. (1985), reported that cows with increased

concentrate consumption during the dry period were at lower risk of LDA (OR=0.3 to 0.4)

and ketosis (OR=0.2 to 0.8), respectively. However, Correa et al. (1990), reported that

cows exposed to diets in concentrates were at increased risk of LDA (OR=4.4) and MF

(OR=2.4). Shaver (1997) recommends that until more data are available, an adequate

prepartum concentrate level should be 0.5% of body weight with an upper limit of 0.75%

of body weight. Pre-calving rations containing energy densities > 1.65 Meal of NEL/kg of

DM are associated positively with risk of LDA. These diets are richer in concentrate than

diets < 1.65 Meal of NEL/kg of DM (Cameron et al., 1998). High concentrate diets, rapid

introduction of concentrate in the immediate pre or postpartum period, rations high in corn

silage or low in crude fiber are factors that affect abomasal motility or enhance gas





35

production (Nocek et al., 1983; Markusfeld, 1986; Trent, 1990). Dawson et al. (1992),

reported that cows fed ground alfalfa hay and concentrate in a TMR in early postpartum

were at higher risk for LDA than were cows fed the standard herd ration of sorghum silage

and concentrate mixed plus loose alfalfa hay. The lack of physical form reduces chewing

activity, ruminal fill, motility and fiber mat formation and increases ruminal VFA

concentration, all of which may affect the etiology of LDA. Concentrate DM can be

increased at the rate of 0.20 to 0.25 kg/d until peak lactation is reached. Concentrates

should be fed three to four times daily. Feeding a TMR to control forage:concentrate ratio

is recommended. A transition group TMR with a higher effective fiber content for early

postpartum cows is also recommended. The importance of physical form of fiber as a risk

factor for LDA is likely to be greatest during the early postpartum period because of the

physiologic and metabolic changes in the transition period. The lack of physical form

reduces chewing activity, ruminal fill, motility, fiber mat formation, and increases ruminal

VFA concentration, all of which may affect the etiology of LDA (Shaver et al., 1986;

Bauchemin, 1991; Mertens, 1992; Muller, 1992; Shaver, 1997; Varga et al., 1998;

Heinrichs et al., 1999).

Different forage program for dry cows are used under field conditions, but data are

limited regarding their impact on the incidence of LDA (Shaver, 1997). Nocek et al.

(1983) evaluated three dry cow forage programs consisting of long hay, hay and corn

silage (50% each one) and corn silage. The incidence risk for LDA was 3.0, 4.3 and 6.3%,

respectively. The incidence risk for ketosis was 9.1%, 6.3 and 6.4%, respectively. Shaver

(1997) recommends that rations with 100% corn silage as forage source should not be fed

to dry cows. The controlled use of corn silage as a component of forage programs for dry

cows may be beneficial.





36

The importance of bunk management practices that limit feed intake in the

etiology of LDA is likely to be greatest during the early postpartum period because of the

coinciding events of the transition period. TMR mixing can alter the physical form of the

fiber in the diet. Excess TMR mixing may grind coarse particles and cause a lack of fiber

physical form. Furthermore, excess particle size can allow the cows to sort the TMR in the

feed bunk which can also cause the same problem (Shaver, 1997; Vargas et al., 1998;

Heinrichs et al., 1999; Melendez et al., 2002; Melendez et al., 2003d).

Right dilatation of the abomasum and abomasal volvulus (RDA-AV)

Right displacement and volvulus are less common conditions constituting 5 to 15%

of the total cases of displacements or volvulus (Trent, 1990).

Cattle with AV are more likely to appear systemically affected than those with

RDA or LDA. Shock, hypovolemia and pain may be present associated with distention

and necrosis of the abomasum, or severe electrolyte and acid-base imbalances, or both.

There is an abrupt decline in milk production. Feces are scant and often dark and

diarrhreal (Fecteau et al., 1999).

Many of the predisposing factors suggested for LDA, such as those that act by

altering motility or promoting gas build up, have been suggested as causative for RDA as

well. Right displacement of the abomasum is more diffusely distributed throughout the

lactation period than are LDAs (Trent, 1990). Both RDA and AV typically result in

hypochloremic, hypokalemic, metabolic alkalosis, paradoxical aciduria and are frequently

associated with hypocalcemia. However these metabolic changes are more pronounced in

AV than RDA (Fecteau et al., 1999).





37

Mastitis

Mastitis is the most common and costly disease of dairy cows. The major

economic loss of all forms of mastitis results from reduced milk production (De Graves

and Fetrow, 1993). Mastitis losses have been estimated at $2 billion per year in the U.S.A

or approximately $200 per cow annually (Fetrow and Anderson, 1987).

Mastitis may be clinical or subclinical. Clinical mastitis is defined as visually

abnormal milk secretion (e.g., clots, flakes, or watery) from one or more quarters. It might

or might not be accompanied by signs of inflammation of the udder tissue (e.g., heat,

swelling, or discoloration of the skin (Kelton et al., 1998). Subclinical mastitis is defined

as the presence of pathogenic microorganisms in the milk and a somatic cell count (SCC)

above 500,000 cells/ml. Reduced milk production associated with subclinical mastitis

accounts for approximately 70% of the economic loss. Treatment costs, culling, and

reduced productivity associated with clinical mastitis are responsible for the remaining

losses (Radostits et al., 1994). There is a straight line negative relationship between a

logarithmic transformation of SCC (LS) and milk production. Estimates of milk

production losses range from 3 to 6% with each unit increase of the LS (Barlett et al.,

1990). Losses associated with the treatment of clinical mastitis have been estimated at

more than $100 per episode (Hoblet et al., 1991).

The reported frequency of clinical mastitis, based on 62 citations from 1982 to

1996 ranged from a lactational incidence of 1.7% to 54.6%. A new case was defined when

abnormal milk in a different or the same quarter followed at least 8 days of normal milk

(Kelton et al., 1998).

The causes of bovine mastitis have been classified as major pathogens and minor

pathogens. The major mastitis pathogens can be further classified as causes of contagious





38

or environmental mastitis. The most common contagious mastitis pathogens are

Streptococcus. agalactiae and Staphylococcus. aureus. Environmental mastitis refers to

infections caused by two categories of organisms, coliform and streptococcal species

(Radostits et al., 1994). There is increasing evidence that, as the contagious pathogens are

progressively controlled in a herd, the incidence of clinical cases caused by coliforms

organisms increases (Jones, 1990).

Mastitis is a multifactorial disease where three major factors are involved: host

resistance, microbial agents, and the environment. The general resistance by the host is

related to genetic predisposition, anatomic characteristics, nutritional status, stage of

lactation and parity (Radostits et al., 1994). Periparturient diseases, such as dystocia,

parturient paresis, RFM and ketosis have all been identified as risk factors for the

subsequent development of mastitis. Cows with MF are 5.0 times more likely to develop

clinical mastitis than normal cows (Curtis et al., 1983).

Lameness

Lameness is one of the most prevalent disease in dairy cattle operations (Shearer,

1996) Lameness is a multifactorial condition and costly disease where many factors have

been identified. Nutrition, genetics or conformation, facilities, environment and hygiene,

behavior and management are the most important risk factors in the etiology of lameness

in cattle (Greenough et al., 1997). As examples, in a study conducted at the University of

Florida, cows that were fed a diet low in prepartum energy and high in postpartum energy

developed subclinical laminitis. This feeding protocol was associated with a significantly

higher rate of rumen acidosis than rations with relatively low postpartum energy

(Donovan et al., in press). In another study conducted in Canada, the heritability of





39

lameness was calculated at 0.16. The genetic and phenotypic correlation between 305-d

milk yield and lameness was 0.24 and 0.04 respectively (Van Dorp et al., 1998).

Prevalence and incidence of lameness have been reported worldwide with a large

variation in the values. Different methodologies have been used and results must be

interpreted with caution. The frequency of lameness, based on 39 citations from 1972 to

1995 ranged from 1.8% to a mean annual incidence rate of 30% (Kelton et al., 1998).

Wells et al. (1993), in Wisconsin and Minnesota, Kaneene and Hurd (1990), in Michigan,

and Miller and Dorn (1990), in Ohio reported an annual incidence of lameness of 6.6, 6.9,

and 5.1 cases per 100 cows at risk, respectively. Throughout the United States, the USDA-

APHIS (1997) showed that the annual incidence of lameness in herds less than 100 cows

was 15.2% and over 100 cows was 18.8%. The region with the highest incidence was the

Northeast (21.2%) and with the lowest incidence was the Southeast (8.6%). In Florida, at

the Dairy Research herd of the University of Florida, the incidence of lameness was

determined to be 51 cases per 100 cows in a year (Shearer and Elliot, 1998).

The cost of lameness is related to treatment cost, losses in milk production,

discarded milk, decrease in dry matter intake, reproductive problems, culling, opportunity

costs and susceptibility to other diseases. A cost of a case of interdigital lameness has been

calculated as $160, a case of digital lameness $392 and a case of sole ulcer $700

(Greenough et al., 1997). Guard (1994) estimated an average cost of a case of lameness to

total approximately $302. Deluyker et al. (1991) reported that lame Holstein cows had a

drop in milk production of 1.7 kg/day during a 4-week period around diagnosis. In Finish

Ayrshire cows, Rajala-Schultz et al. (1999b) found that lameness affected milk

production. In this study, cows with foot and leg disorders experienced milk losses that





40

varied between 1.5 to 2.8 kg/day during the first 2 weeks after the diagnosis. However the

lactational incidence of lameness was as low as 2.1% over a population of 23,416 cows.

Lameness has been described more likely to occur during the first 60 to 90 days of

lactation (Rowlands et al., 1985). Gr6hn et al. (1990a) described a lactational incidence of

lameness of 1.9%, and the median day postpartum of diagnosis of the lameness was 65

days.

Although lameness has not been strongly related with calving-related disorders it

has been considered as a risk factor for some postpartum diseases in dairy cattle. Peeler et

al. (1994) found that cows that experienced dystocia were 1.47 times more likely to

become lame. Furthermore, lame cows were 1.45 times and 1.42 times more likely to

develop mastitis and anestrus than normal cows, respectively. In a logistic regression

model for different periparturient diseases, lame cows were 6.1 times more likely to

develop early metritis than non-lame cows. Finally, in a Florida study, lame cows within

the first 30 days postpartum were 2.63 times more likely to develop ovarian cysts and 0.43

times less likely to become pregnant than non-lame parity-matched cows (Melendez et al.,

2003a).

Culling

Culling is defined as the removal of animals from a dairy production enterprise.

Culling provides the opportunity to progress in genetic improvement, but it can also

represent a substantial loss to the producer (Radostits et al., 1994). Dairy herds can be

expected to cull between 20 and 35% of their cows each year. Genetic progress can be

also increased if the generation interval is shortened, reproductive performance is high,

and calf mortality is low (Radostits et al., 1994).





41

Reasons for culling are death, selling for dairy purposes, low production, poor

reproduction, udder problems, feet and legs problems and miscellaneous (Radostits et al.,

1994). Death was associated with downer cow syndrome. Low production was explained

by low PTA for milk, previous milk production per day and calving season (summer);

poor reproduction was explained mainly by number of services, calf survival, cystic

ovaries and abortion; udder problems were explained by previous milk per day, current

milk per day, mastitis and teat problems; feet and legs problems were mainly explained by

PTA Milk, dystocia and foot or leg problem; miscellaneous reasons were explained by

PTA milk, LDA and mastitis (Milian-Suazo et al., 1989).

In New York Holstein state, older Holstein cows were at much higher risk of

being culled than younger animals. Calving season had no effect on culling. Higher milk

yield was protective against culling. Once a cow had conceived again, her risk of culling

dropped sharply. In all models, mastitis was an important risk factor throughout lactation.

Milk fever, RFM, displacement of abomasum, ketosis and ovarian cysts also significantly

affected culling at different stages of lactation. Metritis had no effect on culling. These

results indicated that diseases have an important impact on the actual decision to cull and

the timing of culling. Parity, milk yield, and conception status are also important factors in

culling decisions (Gr6hn et al., 1998). In Finnish Ayrshire cattle, the farmer's knowledge

of the cow's pregnancy status had a significant effect on culling. The earlier the farmer

knew a cow was pregnant, the smaller was the risk of culling. If a cow had not been bred

at all, her risk of culling was 10 times higher than if she was inseminated once. The effect

of parity decreased when pregnancy status and number of inseminations were added to the

model. Mastitis, teat injuries and lameness had the greatest effect on culling, followed by

anestrus, ovarian cysts and MF. In general, the effect of diseases decreased when





42

reproductive performance was also accounted for in the model. Milk yield had a

significant effect on culling decisions, depending on the stage of lactation. In late lactation

the highest producers were at lowest risk of being culled and the lowest producers had the

highest risk. Milk yield and parity were interrelated in their effects on culling (Rajala-

Schultz and Gr6hn, 1999a; 1999b).

Ketosis and fatty liver

Ketosis is defined as a metabolic disease characterized by high levels of ketone

bodies affecting cattle, sheep and goats. Ketosis affects dairy cows in the period from

parturition to 6 weeks postpartum (Herdt and Gerloff, 1999). There are two types of

ketosis, primary and secondary. Cattle with primary clinical ketosis havea decreased

appetite and elevated serum, milk, urine or breath ketones in the absence of another

concurrent disease (Kelton et al., 1998).

The ketone bodies, acetone, acetoacetate, and B-hydroxybutyrate (BHB), are

formed in the liver during oxidation of fatty acids (Nelson and Cox, 2000). Acetone, a 3

carbon compound, is produced in small quantities and is exhaled to the environment. The

first produced ketone body is acetoacetate (4 carbons), which either is reversibly reduced

to BHB in mitochondria, or is enzymatically or spontaneously decarboxylated to acetone

(Nelson and Cox, 2000).

Ketosis may be clinical or subclinical. Subclinical ketosis is defined as a

preclinical stage characterized by elevated blood ketone body concentrations without

clinical signs such as loss of appetite, hard feces, or dullness (Anderson, 1988; Duffield,

2000; Geishauser et al., 2001).

The lactational incidence rate of clinical ketosis, based on 36 citations from 1979

to 1995, ranged from 1.3% to 18.3%. The median lactational incidence rate was 4.8%





43

(Kelton et al., 1998). Rajala-Shultz et al., (1999) reported in Finish Ayrshire cattle an

overall incidence of 3.3%, however, in some herds, ketosis can be a particular problem

and can affect a large proportion of at-risk cows (Herdt and Gerloff, 1999). Gr6hn et al.,

(1998) reported an overall incidence of 5.0% in New York Holstein cattle. They

determined a range between 4 to 22% among 8 herds as well (Gr6hn et al. (1999).

Ostergaard and Gr6hn, (1999), in Danish cattle determined an incidence of ketosis of

2.0% in primiparous cows and 10.0% in multiparous cows. Al-Rawashdeh, (1999) in

Jordanian dairy cattle determined an overall prevalence of mild ketonemia (0.9 1.7

mmol/1 of BHB) of 22% and severe ketonemia (> 1.7 mmol/1 of BHB) of 3.8%.

More than 90% of subclinical ketosis cases occur in the first and second months

after calving. During this period, approximately 40% of all cows are affected by

subclinical ketosis at least once, although the incidence and prevalence are highest in the

first and second weeks after parturition (Duffield, 2000; Geishauser et al., 2001). In a

Finish study, the odds of contracting ketosis during the first 39 days in milk was higher

than cows beyond 39 days postpartum (Schnier et al., 2002).

Economic losses from ketosis include treatment of clinically ill cows, lost milk

production, increased days open, and increased culling, and were estimated to total

approximately $145 per case (Guard, 1994). Seven studies found short- or long- term

reduction in milk yield associated with clinical ketosis, whereas four reported no losses

(Fourichon et al., 1999). Five studies found losses associated with ketotic status according

to a diagnostic test, whereas two reported no losses (Fourichon et al., 1999). These

discrepancies might be explained by the different statistical methods used to analyze the

effect of the disease on milk production. Rajala-Shultz et al. (1999), using repeated

measures, based on a mixed model with a special parametric structure for the covariance





44

matrices, found a significant negative effect of ketosis on milk production. The milk-

reducing effect started even before the diagnosis of clinical ketosis. Milk losses continued

for at least 2 weeks after diagnosis and the overall losses during the entire lactation ranged

from 126 kg in parity 1 to 535 kg in parity 4 and an average total loss per cow of 353 kg.

Daily milk loss within the first two weeks after diagnosis was 3.0, 4.0, 3.3, and 5.3 kg/d

for parities 1, 2, 3, and 4 or higher, respectively. In parity 4 or higher, the milk loss

continued for the rest of the lactation. This could be an indication that the energy

requirements of the cow were not met. However, healthy cows overall produced less milk

than ketotic cows in the same parity. This might indicate that cows with ketosis are, in

general, higher yielding and that the milk loss often is only temporary. The same

conclusion was observed in New York Holstein cattle when they did a modeling of the

effect of ketosis on milk yield, using a mixed model with four different covariance

structures Gr6hn et al., (1999). By using projected 305-day milk yield, ketosis had no

effect on milk production; but by using a monthly test-day milk as a repeated

measurements and four covariance structures, ketotic cows yielded significantly less milk

per day both before and immediately after diagnosis than did non-ketotic cows. In another

study, Ostergaard and Gr6hn (1999), in Danish cattle, found that ketotic cows had

significantly higher milk yield before the disease that did healthy cows. Multiparous cows

lost between 1.5 and 4.2 kg/day of milk between 1 to 4 weeks after diagnosis of ketosis.

Furthermore, the body weight of cows with ketosis was lower after diagnosis than was that

of healthy cows. In Swiss cattle, a significant decrease in milk production (442 to 654 kg

of energy-corrected milk/305-day period per cow) was associated with acetone or BHB in

excess of threshold values of > 0.40 mmol/L and > 2.3 mmol/L, respectively (Reist et al.,

2003).





45

The effect of ketosis on fertility has been described by Andersson et al., (1991).

They found, in Swedish cattle, that the interval from calving to first service was about 5

days longer in cows with acetone concentrations > 2.0 mmol/L, while the interval to the

last service was shortest at 0.40 to 1.0 mmol/L. The risk for cystic ovaries was markedly

increased (odd ratio = 8.7) in first lactation cows with acetone concentrations > 2.0

mmol/L. In herds with a high incidence of ketosis, primiparous cows had a period 6 days

longer from calving to first service and the period from calving to the last service was 12

days longer than normal cows. Recently, Gillund et al. (2001), found in a Norwegian herd

producing 6,175 kg milk/cow/lactation that 20% of the animals were ketotic as measured

by an Acetone test. The mean days in milk for the first occurrence of ketosis was 29.4.

Cows that did not experience an event of ketosis before first insemination were 1.6 times

more likely to conceive than cows that were ketotic during this early postpartum period.

Ketotic cows had higher BCS than healthy cows before the disease was diagnosed and lost

more BCS than did the latter after ketosis had occurred. Finally, the interval from calving

to conception or number of services per conception did not differ between ketotic and

nonketotic cows. In a Swiss study (Reist et al., 2003), high milk acetone concentration

(>0.4 mmol/L) was associated with 3.2 times higher risk of endometritis. Low plasma

glucose, high serum BHB (> 2.3 mmol/L) and high milk acetone concentrations (> 0.4

mmol/L) during the first week after parturition were indicators of increased risk for ketosis

later during lactation. High BCS (> 3.5; scale 1-5) prior to parturition was associated with

high concentrations of ketones in both milk (acetone > 0.4 mmol/L) and serum (BHB >

2.3 mmol/L). Cows with high BCS prepartum were 3.77 (CI 95%: 1.63-8.71) times more

likely to have milk acetone levels > 0.4 mmol/L and 6.54 (CI 95%: 2.0-21.4) times more

likely to have serum BHB levels > 2.3 mmol/L during the first week postpartum.





46

The effect ofketosis on culling was determined described in Holstein cattle in New

York State by two groups of researchers in two different time periods (Milian-Suazo et al.,

1988; Gr6hn et al., 1998) They found that ketotic cows were more likely to be culled

throughout lactation than were nonketotic cows. However, Gillun et al. (2001) in Norway

found no difference in culling rate between ketotic and nonketotic cows.

During early lactation dairy cows experience a typical negative energy balance

characterized by mobilization of NEFA from adipose tissue (Goff and Horst, 1997b; Herdt

and Gerloff, 1999; Herdt, 2000). This is explained by low DMI at parturition and slower

increase in DMI than in milk production during the early postpartum period. Energy

required for maintenance and milk production exceeds the amount of energy the cow can

obtain from dietary sources (Goff and Horst, 1997b).

Herdt and Gerloff, (1999) establish that distinct metabolic types of ketosis might

exist dependent upon hepatic patterns of NEFA metabolism. Glucose availability is an

important factor in the pathogenesis of clinical ketosis. When glucose availability is very

low, entry of NEFA into a ketogenic liver pathway is favored, but when glucose is high,

esterification and fat deposition are favored (Herdt, 2000). Two factors determine glucose

availability: (1) rate of gluconeogenesis and (2) availability of gluconeogenic substrate.

Rate of gluconeogenesis is increased by the abundance of pyruvate carboxylase during the

early transition period followed by increased abundance of phosphoenol pyruvate

carboxykinase (PEPCK) during the postpartum period (Greenfield et al., 2000). Rate of

gluconeogenesis may be impaired in those cases of ketosis that develop within the first

week postpartum. Evidence suggests that hepatic fat accumulation prior to calving may

interfere directly or indirectly with gluconeogenesis. It has been reported that activity of

PEPCK is decreased in cows that developed fatty liver postpartum (Rukkwamsuk et al.,





47

1999b). In addition, lipid accumulation dramatically decreases ureagenesis with increased

blood ammonia concentrations, which decreases gluconeogenesis from propionate but not

from alanine (Overton et al., 1999).

The entire process of ketogenesis, from adipose mobilization to mitochondrial fatty

acid transport, appears to be affected by the opposing actions of insulin and glucagon,

making the insulin:glucagon (I:G) ratio an important determinant of ketogenesis. Cows in

early lactation have low serum insulin concentrations in the face of normal glucagon

concentrations, resulting in a reduction in the I:G ratio. This low I:G ratio promotes

adipose tissue mobilization and ketogenesis. Ketone bodies, however, appear to stimulate

insulin secretion and therefore may raise the I:G ratio, creating a negative feedback on

ketone body production (De Boer et al., 1985; Herbein et al., 1985; Herdt, 2000). The net

result of these changes and balances is that during periods of negative carbohydrate and

energy balances, ruminants normally have moderately elevated blood ketone body

concentrations and moderately depressed blood glucose concentrations (Herdt and Emery,

1992; Herdt, 2000).

In average in cows, dry matter intake decreases precipitously by 30% on day 1 or 2

before calving and does not recover until 1 to 2 days after calving. Liver triglycerides

(TG) are increased 6 fold by the day of calving (11.8% DM) as compared before calving.

By 4 weeks into lactation the liver TG are 5 fold higher than before calving (10.2% DM).

Plasma NEFA are 5 (1.1 mEq/L) and 1.5 (0.36 mEq/L) times prepartum levels (25 d

prepartum; 0.2 mEq/L) at calving and 35 days postpartum, respectively (Bremmer et al.,

2000). Triglyceride and lipid accumulation in the liver is a much earlier phenomenon than

previously assumed. Elevation of plasma NEFA concentrations starts prior to DMI

depression, on day 5 before parturition. Liver TG infiltration does not occur until the





48

concentration of plasma NEFA is maximized on day 1 after calving (Vazquez-Aflon et al.,

1994). One explanation for this phenomenon is that the rate of hepatic acid esterification

exceeds the rate of TG disappearance via hydrolysis plus the export of newly synthesized

TG as very low density lipoprotein (Grummer, 1993; Hocquette and Bauchart, 1999).

Identification of the biochemical mechanism that limits efficient hepatic oxidation of fatty

acids remains elusive (Goff and Horst, 1997b). One of the theories was that microsomal

TG transfer protein (MPT), which is responsible for transfer of TG into the growing

VLDL particle, might be deficient or inactive in ruminant liver. However, Bremmer et al

(2000) found no correlation between MTP activity, mass, or mRNA with either liver TG

or plasma NEFA on d 2 postpartum. They concluded that MTP probably does not play a

role in the etiology of fatty liver that occurs in dairy cows at calving. Fatty liver can occur

very rapidly. Within 48 hours, hepatic TG levels can increase from less than 5% wet

weight to more than 25% under conditions of extreme adipose mobilization (Gerloff and

Herdt, 1999).

Ketosis cases that develop later during lactation, near the time of peak milk

production, may be of a different metabolic type. Gluconeogenic substrates are simply

insufficient to meet the demands of milk production. This results in high NEFA

concentrations, with a large portion being directed into ketogenesis, rather than

esterification. This kind of ketosis is not responsive to experimental glucagon therapy

(Herdt and Gerloff, 1999). Plasma glucose increased 11.5 and 9.0 mg/dl during week 1

and 2 after glucagon infusions. Nonesterified fatty acids and BHB were not affected (She

et al., 1999; Hippen et al., 1999a). However, in another experiment, Hippen et al. (1999b),

demonstrated that glucagon decreased the degree of fatty liver in early lactation and the

incidence of ketosis after alleviation of fatty liver. Membrane fatty acid composition in





49

liver could impact regulation of NEFA and glucose metabolism in the liver (Drackley et

al., 2001). In cows with fatty liver, content of C 18:1 increased and C 18:2 decreased

relative to prepartum values. In addition, elevated concentration of circulating NEFA is

associated with increased liver concentrations of palmitic, oleic, and linoleic acids, but not

stearic acid (Rukkwamsuk et al., 1999c; 2000).

Methods of diagnosis of ketosis-fatty-liver complex deserve attention. Ketosis is

usually diagnosed based on clinical sings and the level of ketone bodies in urine or milk.

Subclinical ketosis is determined not simply by the presence of ketone bodies, but an

abnormal level of circulating ketone bodies (Duffield, 2000). Ketone bodies can be

measured in blood, urine and milk. In Table 2-5 ketone body concentration from different

studies are reported.

When ketone bodies are measured quantitatively, a defined threshold must be used

to separate normal animals from those with subclinical ketosis (Duffield, 2000). Several

field testing kits have been developed with different sensitivity and specificity values. In

Table 2-6 two studies are summarized.

Subclinical ketosis may start at serum BHB concentrations above 1.0 mmol/L

(10.4 mg/dL) and clinical ketosis at 2.6 mmol/L (27 mg/dL), however these levels are

extremely variable among animals (Duffield, 2000). Serum concentration of 1.4 mmol/L

of BHB (15 mg/dL) or greater in the first 2 weeks postpartum was found to cause a three

fold greater risk for cows to subsequently develop either clinical ketosis or abomasal

displacement. In addition, cows having serum BHB concentrations at or above 2.0

mmol/L (21 mg/dL) within 7 days prior to the first Dairy Herd Improvement (DHI) test

produced over 4 kg less milk on DHI test day (Duffield, 1997). Despite its instability,

blood acetoacetate levels have been used by some authors to identify animals with





50

subclinical ketosis (Duffield, 2000). A threshold of 500 Pmol/L (5.0 mg/dL) of blood

acetoacetate in cows with clinical ketosis has been reported (Baird, 1982). This threshold

would approximate 3.9 mmol/L (40 mg/dL) of BHB.

Potential therapeutic approaches to fatty liver include (1) limiting the fatty acid

supply by limiting adipose mobilization, (2) limiting hepatic fractional extraction (uptake)

of fatty acids, (3) increasing oxidative metabolism of fatty acids, or (4) increasing the rate

of triglyceride secretion from the liver (Herdt and Emery, 1992). Treatment of ketosis is

based on agents that reestablish a normal appetite and restore normal blood concentrations

of glucose and ketone bodies (Herdt and Gerloff, 1999).

Administration of 500 mL of 50% glucose or dextrose solution, intravenously, is a

common therapy for bovine ketosis (Herdt and Emery, 1992). Other therapeutic

alternatives are glucose precursors. They can be administered orally to ruminants. These

include glycerol, propylene glycol and propionate. Glucocorticoid therapy is also

effective. Ketotic cows treated with glucocorticoids are less subject to relapses than are

those treated with IV glucose alone, although relapses can still occur (Herdt and Gerloff,

1999). Dexamethasone and 9-Flurprednisolone acetate at a dose of 1.33 mg/45 kg body

weight and Flumethasone at a dose of 0.33 mg/45 kg of body weight are recommended

(Herdt and Emery, 1992). Insulin, in conjuction with glucocorticoids, may be a more

effective therapy. Insulin is a powerful antiketogenic agent and also suppresses NEFA

mobilization. A long acting form of insulin should be used at a dose of 200 to 300 IU per

animal, repeated as necessary at 24 to 48 hours intervals. Glucocorticoids or other agents

have to be provided to counteract the hypoglycemic effects of insulin (Herdt and Gerloff,

1999).





51

To prevent ketosis-fatty liver complex, fat cows at parturition should be avoided.

Dry cows should be fed a diet to maintain weight, not to lose weight (Gerloff and Herdt,

1999). Cows overfed during the dry period have higher concentrations of plasma

postpartum NEFA as a result of greater lipolysis after parturition. Cows experiencing

more severe negative energy balance results in a high liver TG concentration (Grummer,

1993; Rukkwamsuk et al., 1998, Rukkwamsuk et al., 1999a). Rukkwamsuk et al. (1999a)

demonstrated that cows overfed before parturition, had lower percentages of oleic acid and

higher percentages of linoleic acid than normal cows in liver, but percentages of other

fatty acids were similar. One week after parturition the percentages of palmitic and oleic

acids were higher than one week before parturition. They concluded that the increased

lipolysis after calving increased the hepatic TG concentration and shifted hepatic fatty acid

composition. Kaneene et al. (1997) showed that metabolic events related to negative

energy balance were related to increased risk of metritis and RFM. Higher energy

consumption during the last weeks of the dry period (more grain content of the ration) was

related to reduced disease risk at parturition. Finally, they concluded that serum NEFA

and cholesterol concentrations have potential as indicators of disease risk in dairy cows.

Scoring the body condition of the cows at dry-off and managing the nutritional

program of transition cows is a useful tool to avoid fatty liver development and related

problems (Gerloff and Herdt, 1999). Two adequate body condition score charts are

available for Holstein dairy cattle. One chart was developed by Edmonson et al. (1989)

and the other was developed by Ferguson et al. (1994). It has been demonstrated that high

body condition score at parturition negatively affects reproductive parameters, milk yield

and health of the cows during the current lactation (Butler and Smith, 1989; Domecq et

al., 1997a; Domecq et al., 1997b; Markusfeld et al., 1997; Heuer et al., 1999).





52

Niacin is a vitamin belonging to the B group. As an additive it can reduce lipid

mobilization during early lactation, minimize incidence of ketosis and improve milk yield

and fat synthesis (Hutjens, 1991). In several studies in which 6 to 12 g/day of niacin was

supplemented to dairy cows fed diets with added fat, niacin increased milk production and

concentration of milk fat and protein. Niacin or nicotinic acid has been also suggested as a

treatment or preventive for fatty liver in dairy cows during the prepartum period; but there

is little research evidence to confirm its use as a preventive agent for fatty liver (Gerloff

and Herdt, 1999). Niacin is antilipolytic and may decrease plasma concentrations of

NEFA and BHB (Dufva et al., 1983). Nevertheless, in another study, niacin either alone or

in combination with high dietary non-fiber carbohydrates (NFC), did not influence

lactation performance or metabolic status of cows. Increased dietary NFC significantly

lowered plasma NEFA and BHB concentrations, and significantly elevated liver glycogen

(Minor et al., 1998). In another study, Drackley et al. (1998) determined that nicotinic acid

increased NEFA when it was added to a control diet but decreased NEFA when it was

added to a diet supplemented with fat. Although data seem to be controversial with respect

to niacin in transition cows, the addition of 6 to 12 g/d of niacin to diets during the first 2

to 3 month of lactation has been recommended. Heavier dairy cows (BCS > 3.5) have

shown optimal response; thin cows are less responsive. Niacin should be supplemented 2-

3 weeks before parturiton to 10-12 weeks after parturition, but it should be analyzed on a

cost vs response basis (Hutjens, 1992).

Propylene glycol is a glucogenic compound used to treat ketosis in postpartum

dairy cows (Studer et al., 1993). A high proportion of propylene glycol by-passes the

rumen and is absorbed in the small intestine; the rest is metabolized to propionate.

Ruminal molar percentage of acetate decreased and acetate to propionate ratio also





53

decreased as propylene glycol dose was increased, indicating ruminal conversion of

propylene glycol to propionate (Grummer et al., 1994; Christensen et al., 1997). In liver,

propylene glycol is converted to glucose, primarily via the lactaldehyde pathway and

subsequent oxidation to lactate (Nelson and Cox, 2000).

Recommended doses of propylene glycol have been in the range of 250 to 400 g,

administered orally, twice a day (Herdt and Gerloff, 1999). Toxicity has been seen with

higher doses. Neurological symptoms develop in 2 to 4 hours and resolved by 24 hours

after dosing. Ataxia is typical. Depression and recumbence are also experienced. Serum

and cerebrospinal fluid osmolality is increased. The median toxic dose ofpropylene glycol

in cattle is 2.6 g/kg of body weight (Pintchuk et al., 1993).

Administration of propylene glycol before parturition has demonstrated to decrease

hepatic fat accumulation and ketone bodies formation. One liter of propylene glycol given

orally once daily during the last 10 days prepartum increased plasma glucose in the treated

group before parturition, but glucose levels were similar between treated and control

group, after calving. Plasma NEFA concentration was 403 and 234 itM, and plasma

insulin concentrations were 0.354 and 0.679 ng/ml for control and treated cows,

respectively from 1 to 7 days prepartum. Plasma NEFA tended to be lower in treated cows

at 1 to 21 days post partum. In the treated group, hepatic TG accumulation was reduced 32

and 42% at 1 and 21 days post partum, respectively. Prepartum plasma glucose, NEFA,

BHB and insulin were strongly correlated with liver TG at 1 day postpartum (r = -0.49,

0.45, 0.36, and -0.49, respectively). Milk production and composition through 21 days

postpartum were not different between the treated and control groups (Studer et al., 1993).

Grummer et al., (1994) conducted an experiment to determine the effectiveness of various

doses of propylene glycol in reducing plasma NEFA concentrations during restriction of





54

feed intake in pregnant Holstein heifers 90 days prior to calving. The results showed that

propylene glycol linearly increased glucose and insulin and decreased BHB and NEFA in

blood. They concluded that the dose of 296 ml of propylene glycol was almost as effective

as a dose of 887 ml in reducing lipid mobilization during restricted feed intake. After that,

the same research group (Christensen et al., 1997) demonstrated that administration of

propylene glycol as an oral drench or mixed with concentrate resulted in higher serum

insulin and lower plasma NEFA concentrations than did feeding propylene glycol as part

of a TMR system. Finally, Laranja da Fonseca et al. (1998) found that cows fed 300 mg/d

propylene glycol orally had an increased milk yield of 2.5 kg/day, glucose was similar,

BHB was lower by almost 2 mg/dl and NEFA were lower by 0.031 mEq/L than cows

without the supplement; however these differences were not statistically significant.

Propionate is produced in the rumen after fermentation of starch, fiber, and protein.

It is the major glucose precursor in ruminants that are in positive energy balance and is the

major substrate used for gluconeogenesis, followed by lactate and amino acids. Its

contribution to total glucose production has been consistently measured at 30 to 50%.

Most of the propionate absorbed from the rumen is metabolized in the rumen epithelium

or liver. (Herdt, 1988). A small portion escapes hepatic and rumen epithelial metabolism,

passes into the systemic blood circulation, and is extracted by peripheral tissues (Miettinen

and Huhtanen, 1996).

After being absorbed by the rumen epithelial wall, propionate is transported via

the portal vein to the liver where it is converted to glucose via pyruvate and oxaloacetate.

Propionate decreases the ruminal absorption of acetate and butyrate. Propionate is

incorporated into the Krebs cycle through succinyl coenzyme A and is converted to





55

oxaloacetate and finally to glucose by the liver (Casse et al., 1994). In Figure 2-1 major

energy biochemical pathways in cattle are summarized.

Nutritionally, propionate can be supplied orally in the form of sodium propionate

or calcium propionate. Calcium propionate has been studied as a calcium-energy substrate

for dairy cows (Melendez et al., 2002a). Calcium propionate is a gluconeogenic precursor

as well as a source of calcium, which will be absorbed by the small intestine and may help

to prevent clinical and subclinical hypocalcemia (Goff et al., 1996). Calcium propionate is

a compound poorly fermented by the rumen microorganism. At parturition, calcium

propionate increases blood glucose 24 hrs after its administration, and reduces BHB and

NEFA during the first two days postpartum. Furthermore, calcium propionate increases

blood calcium, reduces the incidence of clinical and subclinical hypocalcemia and

increases milk yield by 3.8 kg/d during the first 2 weeks after calving (Higgins et al.,

1996). In transition cows fed anionic salts prepartum, a calcium propionate (510 g) plus

propylene glycol (400 g) drench did not affect postpartum concentrations of Ca, P, Mg,

glucose, NEFA, or BHB (Melendez et al., 2002a).





56

Table 2-5. Ketone body concentrations for clinical and subclinical ketosis according to
different authors
Reference Class Ketone body and level


Sauer et al., 1989 subclinical blood,ketones >0.9 mmol/L

Anderson et al., 1991 hyperketonemic milk, Ac > 0.4 mmol/L

Gustafsson et al., 1993 hyperketonemic milk, Ac > 2.0 mmol/L

Gustafsson hyperketonemic milk, Ac > 1.4 mmol/L
and Emanuelson, 1996

Shpigel et al., 1996 clinical urine, AcAc > 60 mg/dL

Duffield,et al., 1998a subclinical serum, BHB >1.2 mmol/L

Geishauser et al., 1998 subclinical serum BHB > 1.2 mmol/L

Jorritsma et al., 1998 subclinical milk, BHB 1.0 mmol/L

Al-Rawashdeh, 1999 mild ketonemia serum, BHB 0.9-1.7 mmol/L
severe ketonemia serum, BHB > 1.7 mmol/L

Green et al., 1999 subclinical serum BHB > 1.2 mmol/L

Herdt and Gerloff, 1999 clinical serum, ketones >35 mg/dL
serum, BHB > 25 mg/dL

Geishauser et al., 2000 subclinical seru, BHB > 1.4 mmol/L

Heuer et al., 2001 subclinical milk Acetone > 0.4 mmol/L

Melendez et al., 2002a hyperketonemia plasma, BHB > 10 mg/dL

Reist et al., 2003 subclinical milk, Acetone > 0.4 mmol/L
serum, BHB > 2.3 mmol/L
Ac= Acetone
AcAc= acetoacetate
BHB= P-hydroxy butyrate
Ketones= total ketone bodies (Ac, AcAc, BHB)





57

Table 2-6. Ketone body field test comparison
Product Ketone body Sample sent) spec2) PV(+)(3) PV(-)(4)
Ketolac (T) BHB Milk
> 50 (pmol /L) 91.9 54.9 28.8 97.2
> 100 72.4 89.4 42.7 94.3
> 200 44.8 96.8 73.6 89.8
> 500 17.2 100 100 85.9
> 1000 3.4 100 100 84.0

Ketostix (t) Acetoacetate (*) Urine 4.6 100 100 84.2

Ketocheck (t) Acetoacetate (*) Milk 27.6 100 100 87.5

Utrecht Acetoacetate (*) Milk 42.5 100 100 89.8
powder (t)

Bioketone (t Acetoacetate (*) Milk 33.3 99.8 96.7 88.4

Pink Test #) Acetoacetate Milk
>100 (xjmol /L) 76 93 60 96
>300 38 98 78 92

Ketolac (#) BHB Milk
>50 (p.mol/L) 91 56 21 98
>100 80 76 30 97
>200 59 90 43 94

Uriscan ( Acetoacetate
>500 (pmol /L) Milk 13 100 0 90

Rapignost () Acetoacetate Milk 3 100 0 91
>500 (gmol/L )
(1) Sensitivity,
(2) Specificity
(3) :Positive Predictive Value
(4) :Negative predictive Value
() : Geishauser et al., (1998).
(#) :Geishauser et al., (2000).
(*): Qualitative test (yes/no)





58

Figure 2-1. Pathways of glucose, amino acids and fatty acids metabolism intersect at the
citric acid cycle


C3 Propylene
Glycol Triglyceride


jC6 Glucose


Lactate
Glycerol C16-C18 NEFA's
Alanine
Other Amino
Acids



C2 Acetate


C3


0-2a C4 Ketone Bodies

Aspartate
and other
aminoacids


acid
cycle






Propionate C Glutamate and other
E 1 amino acids
Adapeted from: Herdt, T. H. 1988. Fuel homeostasis in the ruminant. Vet. Clin. North
Am. Food Anim. Pract. 4:213-231.





59

Monensin


lonophores are compounds that collapse the ion gradients across cellular

membranes. Those that are specific for microorganisms can serve as antibiotics. These

compounds shuttle ions across the membranes and kill microbial cells by disrupting

secondary transport processes and energy-conserving reactions (Nelson and Cox, 2000).

Monensin is one of these antibiotics that act as a sodium carrying ionophore. It is a cyclic

peptide that surrounds Na+ and neutralizes its positive charge. The peptide carries Na+

across membranes down its concentration gradient and deflates that gradient (Nelson and

Cox, 2000). The ionophore-cation complex attaches to bacteria and becomes solubilized in

the lipid bilayer of the cell membrane. Once solubilized in the cell membrane, the

complex cation is exchanged for a proton. This culminates in a greater intracellular Na+

concentration, reduced intracellular K+ concentration and lower intracellular pH. Finally,

bacteria are forced to utilize cellular transport systems to dissipate the intracellular H+ and

Na+. This reduces energy reserves, resulting in lowered capability for cell division and

protein synthesis. Cell death comes from acidification of the cytoplasm (McGuffey et al.,

2001).

Monensin was discovered in an in vitro rumen batch fermentation screening

program (Richardson et al., 1976). Monensin enhances propionate production in a high-

roughage and high-grain diets by 49% and 76%, respectively (Van Maanen et al., 1978).

Prange et al. (1978) also demonstrated that monensin enhanced rumen propionic acid

production by 44% in Holstein steers fed a 30% concentrate diet. These studies confirmed

that the early observations of monensin-mediated in vitro rumen propionate production

and in vivo enhancements of rumen molar percentage of propionate are in fact indicative

of elevated in vivo propionic acid production rates. Monensin works by inhibiting





60

hydrogen producing-bacteria such as Ruminococcus and Butyrivibrio and thereby favoring

propionate producers (Van Soest, 1994).

Rumen Microbial Ecosystem

The adult rumen evolved to aid the digestion of grasses. The rumen microbes

hydrolyze the plant celluloses, hemicelluloses, pectins, fructosans, starches and other

polysaccharides to monomeric or dimeric sugars which are further fermented to give

various products such as acetic, propionic, and butyric acids, methane and carbon dioxide

(Hobson, 1997).

Rumen microorganisms are predominantly strict anaerobes. Some oxygen can be

tolerated as long as the fermentation is sufficiently active to facilitate the disposal of

oxygen and the potential (Eh) of the medium remains within normal limits (-250 to -450

mV). Oxygen introduced through feed and water may diffuse across the rumen wall and

affect organisms near the wall, where oxygen serves as an electron acceptor. Facultative

anaerobes consume oxygen and help to maintain the low Eh (Van Soest, 1994).

Rumen microorganisms are represented by bacteria, protozoa and fungi. In Table

2-7, relative volume and number of microbial organisms are shown.


Table.2-7. Relative volumes and number of microbial organisms
Group Number Mean cell Net mass Generation % of total rumen
mL volume (g3) (mg/dL) time microbial mass
Small bacteria 1 x 1010 1 1600 20 min 60-90
Selenomonads 1 x 108 30 300
Large bacteria 1 x 106 250 25
Protozoa 3 x 109 1 x 109 300 8 h 10-40
Fungi 1 x 104 1 x 105 24 h 5-10
Adapted from: Van Soest, P.J. 1994.Microbes in the gut. Pages 253-280 in Nutritional
ecology of the ruminant. Second edition. P. J. Van Soest. Cornell University Press.





61

According to Stewart et al. (1997) the major groups of rumen bacteria are:
- Prevotella species (formerly Bacteroides ruminicola): strictly anaerobic, gram-

negative bacteria. They degrade starch, xylans and pectins. Fermentation products

include acetate, succinate and propionate. They are sensitive to monensin, but

mutations to resistance occur readily.

- Ruminobacter (Bacteroides) amylophilus: Rare, but it appears to be the predominant

starch digester. It also possesses significant proteolytic activity. Major fermentation

products are acetate and succinate.

- Fibrobacter (Bacteroides) succinogenes: One of the most widespread celluloytic

bacteria of the rumen. Major fermentation products are acetate and succinate.

- Succinovibrio dextrinosolvens: Gram-negative rod ferments pectins and dextrins.

Major products are acetate and succinate.

- Succinimonas amylolytica: Gram-negative bacteria associated with starch digestion.

Major products are acetate and propionate.

- Spirochaetes: The account for 1% to 6% of the total viable bacteria count in bovine

rumen liquor. The rumen spirichaetes are predominantly treponemes. They ferment

pectins and small sugars. Major products are acetate, formate and succinate.

- Anaerovibrio lipolytica: Gram-negative rods that hydrolyze lipids and ferment

fructose, glycerol and lactate. Major products are propionate, succinate, lactate, CO2

and H2.

- Selenomonas ruminantium: This gram-negative curve rod constitutes 22-51% of the

total viable count. Some strains ferment starch, others ferment small sugars, glycerol

or lactate. Another important activity of S. ruminantium is the decarboxylation of

succinate to propionate. Major products are lactate, propionate and acetate. They

appear to be relatively resistant to monensin.





62

- Mitsuokella multiacidus: These gram-negative bacteria were previously regarded as

Bacteroides ruminicola. They produce lactate as a major fermentation product.

- Megasphaera elsdenii: These gram-negative cocci are mainly found in the rumen of

young animals. They ferment small sugars, glucose, fructose, and mostly lactate. This

bacterium is thought to play a major role in the production of branched-chain volatile

fatty acids. End products are butyrate, propionate, isobutyrate, valerate, CO2 and some

H2.

- Ruminococcus species: Represented mostly by two species involved in plant fiber

degradation in the rumen, R. albus and R. flavefaciens. They are gram-positive

cellulolytic bacteria. Major products are acetate, succinate, CO2 and H2. They are

extremely sensitive to monensin.

- Streptococcus bovis: Gram-positive cocci capable of very rapid growth. It is the most

rapidly acting amylolytic bacteria of the rumen. The major fermentation product is

lactate. Minor products are acetate, formate and CO2. Lactate production increases by

the conditions that develop when large amounts of starch are fed and the rumen pH

falls.

- Butyrivibrio fibrisolvens: Gram-negative, strictly anaerobic rods. Predominant bacteria

in the rumen of cows fed on a variety of different rations, ranging from alfalfa hay to

grain mixture. It is the major butyrate-producing species in the rumen, but they also

produce acetate, succinate, CO2 and H2. They ferment cellulose and xylans.

- Lachnospira multipara: Gram-positive curve rod degrading pectins. Major products

are formate, acetate, lactate, CO2 and H2.





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- Clostridium species: They are not predominant bacteria, but 3 cellulolytic species have

been described. They ferment starch and cellulose. Major products are formate,

butyrate, acetate, CO2 and H2.

- Eubacterium species: Gram-positive short rods. They produce acetate from CO2 and

H2. They account for 5% of the total bacteria in rumen.

- Lactobacillus species: More common in young animals fed with milk and in older

animals fed concentrates. Major product is lactate.

- Bifidobacterium species: They catabolize glucose, resulting in the production of acetic

and lactic acids.

- Non-sugar-fermenting bacteria: They are a large population of rumen bacterial species

that utilize organic compounds other than sugar. Major products are succinate,

propionate and acetate.

- Large bacteria: Little is known about this family. They ferment small sugars and

produce acids and gases.

- Methanogens: These organisms are not bacteria; in several respects they more closely

resemble eukaryotes. Methane is formed from CO2 and H2, but other potential

substrates include formate, acetate, and methanol.

According to Williams and Colleman (1997), in addition to the bacteria in the

rumen there are many larger organisms (5-250 tm) belonging to the group of protozoa.

The largest most obvious and most important protozoa are the ciliates, of which there are

two groups, holotrichs and entodiniomorphs. Despite the metabolic diversity of the ciliates

and their large biomass in the rumen, the protozoa, unlike the bacteria, are not essential for

the survival and development of the host ruminant. However, rumen fermentation is more

stable when an active ciliate population is present. Ciliates are selectively retained in the





64

rumen. Whereas bacteria flow completely from the rumen, protozoa flow only 20-40% of

that rate; however, approximately 25% of the microbial protein available to the host is

protozoal in origin. Approximately 1/4 to 1/3 of fiber degradation in the rumen is

protozoal. Bacterial ingestion and proteolysis by the protozoa decreases the efficiency of

nitrogen utilization by the host. An important consequence of the protozoa is their ability

to ingest starch and soluble sugars, preventing the alternative rapid bacterial fermentation

to lactic acid. They have an important role in regulating rumen lactate metabolism and in

preventing lactic acidosis. Some protozoa have been demonstrated to be sensitive to

monensin.

According to Van Soest (1994) and Orpin and Joblin (1997), yeast and anaerobic

fungi have long been known to be normal inhabitants of the rumen. Fungi are much more

particle-associated and involved in fibrolytic digestion than are protozoa. Fungi secrete a

more soluble cellulase complex than do rumen bacteria, and the mechanism of enzyme-

substrate interaction differ as well. The fungi produce volatile fatty acids, gas, and traces

of ethanol and lactate.

Volatile Fatty Acids

Volatile fatty acids (VFA) are end products of anaerobic microbial metabolism in

the rumen (Nagaraja et al., 1997). Major VFA in descending order of abundance are

acetic, propionic, butyric, isobutyric, valeric and isovaleric. The proportion of acetic,

propionic, and butyric acids can be markedly influenced by diet and the status of the

methanogen population in the rumen. Lactic acid is important when starch is part of the

diet, and is itself fermented to acetate, propionate, and butyrate (Van Soest, 1994). Rumen

concentrations of VFA are regulated by a balance between production and absorption

whereby increased production rate induces higher VFA concentrations (Nagaraja et al.,





65

1997). Volatile fatty acid production rates vary diurnally as a consequence of eating

pattern, therefore VFA rumen concentrations and pH vary as well. Fermentation peaks

about 4 hours after feeding on a hay diet but occurs sooner if the diet contains some

concentrate. The major factor affecting VFA absorption is their concentration (Van Soest,

1994). In general, acetate concentration in the rumen ranges between 60 to 70 mmol/L,

propionate between 18 to 25 mmol/L and butyrate between 10 to 15 mmol/L (Nagaraja et

al., 1997)

Monensin and Rumen Fermentation

The most consistent and well-documented fermentation alteration observed upon

monensin feeding is the increased molar proportion of propionic acid with a concurrent

decrease in the molar proportion of acetate and butyrate in the VFA produced in the rumen

(Richardson et al., 1976). Addition of cellobiose and monensin to an in vitro fermentation

batch demonstrated that total concentrations of volatile fatty acids were increased.

Monensin treatment increased propionate and total VFA concentrations and decreased L-

lactate concentrations. A lag in organic acid utilization was observed in fermentations,

which was most likely due to bacteria that were resistant to monensin and that

preferentially utilized soluble nutrients other than organic acids (Callaway and Martin,

1997).

The increase in rumen propionate is accompanied by a reduction in the amount of

methane produced in the rumen (Nagaraja et al., 1997). Ionophore antibiotics are not

inhibitors of methanogenic bacteria. They are believed to reduce precursors of

methanogenesis (H2, Co2 and formate) (Russel and Martin, 1984); however monensin

inhibits methanogenesis from formate, probably as a result of inhibition of nickel uptake





66

by methanogenic bacteria and increased heat increment and the apparent digestibility of

metabolizable energy (Wedegaertner and Johnson, 1983; Oscar and Spears, 1990).

Monensin affects nitrogen metabolism by decreasing ammonia production in

cattle fed a forage -based diet. This might be a result of reduced proteolysis, degradation

of peptides and deamination of amino acids in the rumen (Yang and Russell, 1993;

Nagaraja et al., 1997). Under monensin, the specific activity of ammonia production by

mixed ruminal bacteria is decreased by more than 30% and this decrease corresponds to

about a 10-fold decrease in the numbers of bacteria that ferment peptides and amino acids

as an energy source for growth (Yang and Russell, 1993). Ionophores have also been

demonstrated to decrease ruminal urease activity (Starnes et al., 1984).

Monensin prevents lactic acid build-up in the rumen by its selectivity towards

gram-positive bacteria. The major lactic acid-producing bacteria (S. bovis and

Lactobacillus ssp.) are inhibited while ruminal lactic acid-fermenting bacteria (gram-

negative) are unaffected (Nagaraja et al., 1997). This effect was not repeated in a more

recent study that used a monensin controlled-release capsule or a premix in dairy cattle

(Mutsvangwa et al., 2002).

Monensin also decreases the incidence of frothy bloat as a result of reduction in

microbial slime and gas production (Lowe et al., 1991; Nagaraja et al., 1997). Monensin

reduces or modulates feed intake in both grain-fed and forage-fed cattle, probably due to

low palatability or decrease in rumen turnover rate of liquids and solids, and consequently

increase of ruminal fill (Stock et al., 1995; Nagaraja et al., 1997).





67

Table 2-8. Susceptibility and resistance ofruminal bacteria to monensin.
Species Gram Cell wall Monensin
reaction type susceptibility
Hydrogen and formic acid producers
Lachnospira multiparus + + (0.38)a
Ruminococcus albus + + (0.38)
Ruminococcus flavefaciens + + (0.38)
Butyric acid producers
Butyrivibriofibrisolvens + + (0.38)
Eubacterium cellulosolvens + + + (0.38)
Eubacterium ruminantium + + + (0.38-1.5)
Lactic acid producers
Lactobacillus ruminis + + + (1.5-3.0)
Lactobacillus vitulinus + + + (0.38-1.5)
Streptococcus bovis + + + (0.38-12.0)
Ammonia producers
Clostridium aminophilum + NDb ND
Clostridium sticklandii + + ND
Peptostreptococcus anaerobius + + ND
Succinic and propionic acid producers
Anaerovibrio lipolytica (>48.0)
Fibrobacter succinogenes (>20.0)
Megasphaera elsdenii (>48.0)
Prevotella ruminicola (>48.0)
Ruminobacter amylophilus (>48.0)
Selenomonas ruminantium (>48.0)
Succinimonas amylolytica (>48.0)
Succinivibrio dextrinosolvens (>48.0)
Methane producers
Methanobrevibacter ruminantium NAC NA (>20.0)
Methanobacteriumformicium NA NA (>20.0)
Methanosarcina barkeri NA NA (>20.0)
Adapted from: Nagaraja, T.G., C.J. Newbold, C.J. Van Nevel, and D.I. Demeyer. 1997.
Manipulation of ruminal fermentation. Pages 523-632 in The rumen microbial ecosystem.
Second edition. P.N. Hobson and C.S. Stewart. Blackie Academic & Professional.
a Values in parenthesis are minimum inhibitory concentrations in gtg ml"'
b Not determined
c Not applicable





68

Citrus Pulp and Pectin Fermentation
Citrus pulp is an energy concentrate by-product produced in subtropical regions,

of which south central Florida remains the largest area of production (Arthington et al.,

2002). Citrus pulp is composed of pectin, that is galacturonic acid, arabinose, galactose,

and rhamnose, which are not digested by mammalian enzymes, but can be rapidly

fermented by ruminal microbes (Hall, 1997).

Pectin can inflate ADF values when rich-pectin feeds, like citrus pulp, are

analyzed by using traditional laboratory methods. Plant carbohydrates can be divided into

the neutral detergent fiber fraction (NDF) and the neutral detergent soluble carbohydrate

fraction (NDSC). The NDF fraction is composed of hemicellulose and cellulose. The

NDSC fraction is composed by organic acids, sugars, starches, fructans, pectin and 3-

glucans. Fructans, pectin and B-glucans form the neutral detergent soluble fiber fraction

(NDSF). This fraction can be determined by a simple method that gives good precision

(Hall et al., 1997; Hall et al., 1998). Citrus pulp contains about 34.5 % of NDSF, while

forages contain between 12 to 22% (Hall et al.,.1997).

From in vitro fermentation experiments citrus pulp gives the lowest acetate to

propionate ratio and the highest propionate molar proportion as compared with beet pulp,

soybean hulls, mature and immature alfalfa stems, and mature and immature alfalfa leaves

(Hall et al., 1998).

Diets based on citrus pulp consistently demonstrate an increase in milk fat

content and milk urea nitrogen, but not in milk yield, when compared with diets richer in

starch (Belibasakis and Tsirgogianni, 1996; Leiva et al., 2000). Although higher levels of

milk fat have been suggested to be related to a higher production of acetate in the rumen,

molar proportion and concentration of acetate and propionate did not differ between

experimental diets containing citrus pulp and corn, respectively (Leiva et al., 2000).





69

Since pectin-fermenting bacteria (Prevotella spp, Fibrobacter succinogenes,

Succiniovibrio dextrinosolvens) are gram-negative monensin-resistant bacteria (Nagaraja

et al., 1997; Stewart et al., 1997), it is reasonable to assume that in dairy cows fed

monensin and diets rich in citrus pulp ruminal pectin fermentation should not be altered

significantly.

Monensin, Calving-Related Disorders, Reproductive and Productive Responses

Ionophores have been available for use in food animals for over 20 years,

however, limited information is available about their effects in lactating dairy cows

(Duffield, 1997). During the last 5 years, more information about the use of monensin in

dairy cattle has been developed. Mostly this research has been conducted in Canada and

Australia. Monensin is not allowed for use in lactating dairy cattle in the United States.

One of the earliest reports showing the positive effect of monensin on ketosis

demonstrated that monensin at a rate of 250 mg per cow per day, successfully controlled

an outbreak of ketosis in 18 Friesian cows (Rogers and Hope-Cawdery, 1980). In a well-

designed study conducted in Canada by Sauer et al (1989), cows of second lactation or

older were gradually introduced to a monensin-containing concentrate 1 week prepartum

and fed complete diets containing 0, 15 or 30 g monensin/ton of dry matter for 3 weeks

postpartum (low and high monensin). Acetate molar proportion decreased, while

propionate increased significantly between controls and the high monensin-treated group.

The acetate to propionate ratio was 2.49 and 1.46 in the control and high monensin

groups, respectively. Monensin also decreased the levels of serum BHB in cows in the

high monensin group compared with the control group (3.9 mg/dL vs 7.2 mg/dL,

respectively). Milk production was similar, but milk fat was lower in the high monensin

group than the control group (3.71% vs 4.12%, respectively).





70

Monensin primarily has been used as a powder mixed with concentrates.

Recently a capsule of slow release of monensin has been developed. In Australia, Lean at

al. (1994) and Abe et al. (1994) conducted two studies using a slow-release intraruminal

capsule containing 32 g of sodium monensin. Since that time, capsules have been

available in Australia and Canada.

Lean at al. (1994) postulated that feeding sodium monensin may result in

additional glucose flux and reduced risk of reproductive failure associated with

hypoglycemia and that monensin feeding would increase milk and milk solid yields. They

used a total of 1061 lactating dairy cows from six different herds and management. One

herd was fed a total mixed ration, three were fed on pasture and supplemented

concentrates and two herds were primarily fed on pasture. Capsule was administered

within 7 days of calving. Conception rate at first service, days to first estrus and calving-

to-conception interval did not differ significantly between untreated and monensin-treated

cows. Monensin treatment significantly increased milk yield only in one herd. Milk fat

and protein were not significantly affected by treatement. Abe et al. (1994) wanted to

evaluate the effects of a sodium monensin controlled-release capsule on metabolic

responses and on resumption of ovarian cyclicity. Cows treated with monensin had

significantly lower plasma BHB and tended to have higher plasma glucose than untreated

cows. Treatment did not significantly influence plasma NEFA, urea nitrogen or

cholesterol concentrations. Time to first estrus was significant shorter for untreated than

for treated cows, however inclusion of BCS immediately after calving as a covariate

resulted in non-significant differences. Time to first ovulation did not significantly differ

between groups.





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In another Australian study, where the controlled-release capsule of monensin

was given at 50 7 days before expected parturition, Stephenson et al. (1997) found that

treated cows (n=12) had lower plasma concentrations of glucose, NEFA and BHB than

control cows (n=12) before calving. However, no significant differences in plasma

concentrations of glucose, NEFA and BHB were found between groups after parturition.

Cows were fed on pasture and supplemented concentrate at a level of 4 to 10 kg/cow/day.

Later, Beckett et al. (1998) conducted a randomized clinical trial including 1109

cows from 12 Australian dairy herds. They postulated that treatment administered as an

intraruminal bolus 40 days before expected parturition and 50 days after calving would

reduce the risk of postpartum reproductive failure, reduce the incidence of common

periparturient clinical disorders, and increase the production of milk and milk solids.

Treatment did not significantly alter any reproductive outcome; 54.5% of treated cows and

58.2% of control cows were pregnant at first service. Treatment with monensin did not

reduce the risk of dystocia, ketosis, milk fever, infectious diseases, metritis, RFM,

lameness, or abortion nor did treatment increase the risk of mastitis. However, milk

production was increased in treated cows (0.75 L/d) after adjustment for herd. Milk fat

yield was similar between groups.

In New Zealand, cows from 3 herds fed on pasture received a controlled-release

capsule of monensin 1 month prior to the start of artificial insemination. The objectives of

the study were to determine whether treatment with controlled-release capsules of

monensin improved milk production and reproductive performance of cows calving in

spring and fed pasture. Treated cows produced more fat, protein and milk per day (19.1 vs

17.7 L) during the second month of monensin activity than control cows. Overall

pregnancy rate to first and second service were not different between treated and control





72

cows. Blood urea nitrogen was elevated in the treatment group and NEFA were elevated

in the 2nd and 3rd month after treatment. There was no significant effect on blood

concentrations of glucose, albumin, or BHB, except in the 4h month where BHB was

higher in the treated group (1.75 mmol/L) than control group (1.35 mmol/L) (Hayes et al.,

1996). According to Duffield (2000), these levels are indicative of subclinical ketosis

(BHB > 1.0 mmol/L), which might suggest that cows under those conditions were exposed

to marked energy imbalances.

In Italy, Ramanzin et al. (1997) using 4 primiparous Holstein cows in a Latin

square design with two forage to concentrate ratios (70:30 and 50:50, respectively) and

two concentrations of powder sodium monensin (0 and 300 mg/d/cow, offered

postpartum) determined that monensin tended to depress feed intake and milk fat content

without affecting milk production. Ruminal propionate percentage was increased more by

the addition of monensin to the low forage diet that to the high-forage diet. Serum urea

and NEFA concentrations tended to decrease when monensin was added to the low forage

diet.

In England, Phipps et al. (2000) conducted two experiments to examine the effect

of monensin on feed intake and milk production in Holstein-Friesian cows. In experiment

1, they used 60 lactating cows during weeks 7 to 26 of lactation. There were 4 treatment

groups receiving a 1 kg/d supplement containing 0, 150, 300 or 450 mg of monensin. Dry

matter intake was not different among groups, however milk production was higher in the

groups receiving 150, 300 and 450 mg/d of monensin. Percentage, but not absolute

content (g/d), of fat and protein were lower in the treated groups than in the control group.

In the second experiment, they used 69 multiparous and 29 primiparous Holstein cows

that received either 0 or 300 mg/d of monensin. Cows were evaluated during 2 lactations.





73

Cows evaluated during the second lactation also started to receive monensin 3 weeks

before expected parturion. Milk production was not different between groups in both

lactations, however percentage of fat and protein milk was lower in treated cows than

control cows in both lactations. The use of monensin before calving decreased the BHB

concentration in weeks 2 to 4 and 6 to 8 postcalving. Levels of BHB were between 380

and 524 umol/L, values lower than the cut-off for subclinical ketosis (Duffield, 2000).

Monensin also reduced acetoacetate values in weeks 2 to 4 and increased glucose values

in weeks 2 to 4 and 6 to 8 postpartum. However, there was no difference in these

metabolites in weeks 10 to 12.

In another European study, Van der Werf et al. (1998) in the Netherlands

examined the efficacy of premix monensin on milk production and feed efficiency during

the first 20 weeks of lactation. In a first experiment they used 64 Holstein cows assigned

to 4 groups that received 0, 150, 300, or 450 mg/day of monensin mixed with concentrates

from 5 to 24 weeks postpartum. In a second experiment they utilized 58 Holstein and 22

Jersey cows that were allocated either to a control group or to a treatment group that

received 300 mg/day of monensin from 5 to 36 weeks postpartum during to consecutive

lactations. In trial 1 monensin caused a decrease in fat content for cows fed 450 mg/day.

Feed intake of treated cows tended to be lower than controls. Body weight gain was higher

for cows fed 450 mg/day of monensin. Acetoacetate, BHB and glucose did not differ

among groups. In trial 2, monensin increased milk production during the first lactation in

Holsteins, but not in Jersey cows. At day 56 of treatment a larger decrease in acetoacetate

and BHB and an increase in glucose were found for the treatment group. During the

second lactation acetoacetate and BHB were significantly lower and glucose was higher

for cows in the treatment group than controls. Productive responses were not different.





74

During the last 5 years the group from University of Guelph, Canada, has

developed a strong line of research in transition dairy cows using a controlled-release

capsule of monensin starting in the prepartum transition period. The overall research

project used 1010 dry cows and pregnant heifers from 25 dairy farms near Guelph,

Ontario, Canada. The monensin capsule was inserted 3 weeks prior to expected calving

and released 300 mg of monensin during approximately 95 days. The first report

determined that monensin treatment significantly reduced serum BHB concentrations

between 150 and 200 pmol/L at week 1, 2 and 3 postpartum. Treatment also significantly

raised serum glucose concentrations during weeks 1 and 2 of lactation, with an increase in

serum glucose concentrations of 0.16 to 0.22 mmol/L. Monensin treatment significantly

reduced aspartate aminotransferase activity during the entire postcalving period as well,

especially in weeks 2 and 3 postpartum. In addition, monensin reduced the loss of BCS

after parturition. No significant treatment effects were found for calcium, phosphorus, or

total protein (Duffield et al., 1998a). The authors also reported that incidence and

prevalence of subclinical ketosis (BHB > 1200 Lmol/L) were significantly reduced by

about 50% (odds ratio= 0.46) by monensin treatment. In addition, monensin was also

significant in reducing the incidence of subclinical ketosis by using two other serum

thresholds (1400 and 2000 p.mol/L). The prevalence of subclinical ketosis was also lower

in the treated group than control cows when milk ketone tests were used as the diagnosis

method (Duffield et al., 1998b).

In a third report, the authors evaluated the effect of monensin on milk production

and milk components at the first three Dairy Herd Improvement Association (DHIA) tests.

Treatment with monensin increased milk production, but this effect was dependent on

body condition score prior to calving. Cows with a BCS < 3.0 (scale 1-5) did not have





75

increased production in response to monensin treatment. Cows with a BCS between 3.25

and 3.75 produced significantly more milk at the second DHIA test (0.85 kg), and cows

with a BCS > 4.0 produced significantly more milk than did controls for all three DHIA

tests (1.25 kg/d) Treatment with monensin had no significant effect on either milk fat or

milk protein percentage (Duffield et al., 1999a). After this, Duffield et al., (1999b)

determined the effect of a monensin-controlled release capsule on cow health and

reproductive performance. They found, in 503 cows receiving the bolus, that monensin

significantly reduced the incidence of displacement of the abomasum (OR=0.41-0.84) and

multiple illnesses (OR=0.38-0.89), defined as having more than one of retained placenta,

milk fever, metritis, endometritis, mastitis, ketosis, displaced abomasums, digestive,

lameness, respiratory disease, or other disease. Treatment was not statistically significant

in reducing the incidence of clinical ketosis (P=0.11), defined as reduced feed intake,

testing positive in a milk ketone test, and the absence of any other disease. Treatment with

monensin had no significant effect on the interval from calving to first service. Similarly,

treatment had no effect on the interval calving to conception. Service per conception and

conception rate at first service did not differ between groups as well.

In an effort to complement results on the prevention of subclinical ketosis in

lactating dairy cows, Green et al. (1999) designed an experiment to study subclinical

ketosis in periparturient dairy cows and the antiketogenic effects of monensin. In this

study, the authors induced subclinical ketosis (n=41) through 10% feed restriction and

blood BHB quantification (using a blood BHB threshold of 1200 pmol/L). No treatment

effects were found for daily milk production, milk components, dry matter intake, net

energy balance, body weight and BCS. Rumen pH for the postpartum period was higher

for cows treated with monensin. Cows treated with monensin had lower concentration of





76

rumen butyrate. The rumen concentrations of acetate and propionate were not different.

Mean serum BHB concentration did not differ between groups during the 2 week

postpartum period, although cows treated with monensin tended (P < 0.06) to have a lower

concentration than controls. During the 6 week postpartum period cows treated with

monensin had significantly lower BHB concentration than controls. Control cows had a

serum BHB concentration that exceeded 1200 gmol/L by week throughout the postpartum

period. They concluded that monensin maintained energy-related blood metabolites,

reduced the occurrence of subclinical ketosis, and improved rumen health in the early

lactation dairy cow.

The effect of a monensin controlled-release capsule on diet digestibility and

nitrogen utilization was investigated in 16 multiparous dairy cows between approximately

10 and 3 days precalving and 3 and 9 days postpartum. Monensin decreased significantly

fecal nitrogen output and improved nitrogen balance. The control group lost on average

77.8 g/day of nitrogen, compared with 44.9 g/day in the treated group. The positive effect

of monensin on NDF and ADF digestibility precalving and nitrogen digestibility

postcalving was also significant (Plaizier et al., 2000).

The same Canadian group, studied the effects of monensin administered either as

a controlled release capsule or as a premix, on attenuating grain-induced subacute ruminal

acidosis and on ruminal fermentation characteristics in Holstein cows receiving a total

mixed ration (Mutsvangwa et al., 2002). They used six multiparous Holstein cows in a

two-treatment, two-period crossover design with 6 week periods. Experiment 1 used a

monensin bolus and experiment 2 used a monensin premix. Authors induced subacute

rumen acidosis between day 22 and 32 of each experimental period, by restricting feed

intake to 85% and substituting 15% of the diet with grain pellets of wheat and barley.





77

Neither monensin bolus nor premix had an effect on ruminal pH and VFA concentrations

under conditions of subacute rumen acidosis.

Since monesin is not allowed in lactating dairy cattle in the United States, only a

few studies have been conducted in this country. These studies have been under

supervision of the Food and Drug Administration. One of these studies was conducted to

evaluate the effects of monensin on nitrogen metabolism and the Cornell Net

Carbohydrate and Protein System (CNCPS) in cows fed fresh forage diets. Thirty Holstein

cows in mid lactation (8 with ruminal fistulas) where gradually introduced to a fresh

forage diet. Fifteen cows each were allocated to a control and a treatment group that

received 350 mg/cow per day of monensin in the P.M. concentrate feeding. A 7 day fecal

and urine collection period and a 3 day rumen sampling period were conducted with the

fistulated cows. Monensin increased milk production by 1.85 kg/d. Ruminal ammonia and

the acetate to propionate ratio decreased with the addition of monensin. Monensin

decreased fecal N output, and increased apparent N digestibility by 5.4%. It was

concluded that monensin has the potential to increase the efficiency of N utilization in

dairy cows fed fresh forage and to decrease fecal N excretion. The results suggest that

monensin spared amino acids from wasteful degradation in the rumen (Ruiz et al., 2001).

Transition Cow Feed Management

Feeding a cow during the transition period is a challenge due to the nutritional and

physiological changes that occur during this period (Grummer, 1999). Just recently

nutrient requirements for transition dairy cows have been defined (NRC, 2001). This

information was possible based on research that has been abundant during the past five

years. There have been tremendous changes in how to approach dry cow nutrition,

particularly in the areas of dry matter intake, protein and energy requirements, metabolic





78

diseases and the most effective ways to group and manage dry cows. Critical physiologic

events that have to be targeted during the transition period include adaptation of the rumen

to the higher energy diet that will be fed in early lactation, maintenance of normal blood

calcium concentration, a strong immune system throughout the peripartum period, and

maintenance of a slightly positive energy balance up to the time of calving (Goff and

Horst, 1997b; Oetzel, 1998).

Energy
Energy balance of a transition cow is determined by subtracting energy

requirements for maintenance and gestation from energy intake. During the transition

period, feed intake is decreasing at a time when energy requirements are increasing due to

growth of the conceptus. Consequently, to maintain the energy balance the energy density

of the diet should increase (Grummer, 1999). In Figure 2-2 the net energy of lactation

(NE1) needed in diets of cows and heifers to meet requirements for maintenance and

gestation during the transition period is shown. Heifers need higher dietary energy density

due to lower feed intake and the additional energy requirements to support growth (NRC,

2001).

Increasing energy density may stimulate papillae growth and increase acid

absorption from the rumen, adapt the microbial population to higher starch diets, increase

blood insulin, decrease fatty acid mobilization from adipose tissue and increase dry matter

intake (Grummer, 1999).

Grain has to be introduced to the cow's ration for at least 3 weeks before the due

date and for heifers this should be 5 weeks. The energy density should be between 1.56-

1.62 Mcal/kg of NE1 (NRC, 2001).





79

















2.5

2.25 -

2-


1.75 -
S1.5
1.25 1.25A--A-A--A-A|



0.75 -- Heifer (-mammary)

0.5 ---- Heifer (+mammary)

0.25 -- Cow (- mammary)
-A- Cow (+ mammary)
0 1
21 19 17 15 13 11 9 7 5 3 1
day relative to calving

Figure 2-2. Estimated energy density required in diets of transition cows to meet
requirements for maintenance and gestation according to NRC,
(2001)





80

Increasing dietary NFC or decreasing NDF during the transition period stimulates

DMI. When energy density of the diet increased from 1.3 to 1.54 Meal ENI/kg DM and

crude protein increased from 13 to 16% at about 3 weeks prior to calving, DMI increased

in 30% (Emery, 1993). Plasma fatty acids decreased from 346 to 228 .tM and liver

triglyceride decreased from 15 to 9 mg/g wet tissue (Dyk et al., 1995; VandeHaar et al.,

1995). In another study, cows fed high NFC (40-42%) diets consumed more dry matter

during the prepartum period, had lower plasma NEFA, had reduced BHB level and there

was a reduction in liver triglyceride (Minor et al., 1996). Finally, VandeHaar et al. (1999)

found that increasing nutrient-density of prepartum diets did not decrease DMI. There

were less NEFA in plasma (176 ws. 233 pM) and more insulin-like growth factor-I (472

vs. 390 ng/ml plasma) during the last two weeks prepartum and less triglyceride in liver at

parturition (0.9 vs. 1.5% wet tissue basis). They concluded that increasing the energy and

protein density up to 1.6 Mcal of NEl/kg and 16% CP in diets during the last month before

parturition improves nutrient balance of cattle prepartum and decreases hepatic lipid

content at parturition.

Protein

The protein concentration needed in the diet to meet requirements for maintenance

and gestation during the transition period is shown in Figure 2-3.

The results about crude protein levels in precalving cows are controversial. Curtis

et al. (1985) indicated that feeding protein above NRC (1989) during the final 3 weeks

prepartum decreased the risk of RFM and uncomplicated ketosis. Mahanna (1997)

recommends increasing CP to 14.5% 15.5% to begin preparing for high DMI during the

lactation phase. However, Santos et al. (1999a,b) found precalving heifers fed CP up to

14.7% benefited with higher milk yield and milk fat production, but precalving cows did





81

not benefit from a diet with more than 12.7% CP. In other studies, feeding higher protein

during the transition period reduced feed intake or milk yield postpartum (Crawley and

Kilmer, 1995; Donkin et al., 1998).






20



16 -



12 AAA


U
8 -
-- Heifer (-mammary)
--- Heifer (+mammary)
4 Cow (-mammary)
--- Cow (+ mammary)



21 19 17 15 13 11 9 7 5 3 1

day relative to calving


Figure 2-3. Estimated protein density required in diets of transition cows to meet
requirements for maintenance and gestation according to NRC,
(2001)





82

Minerals and Vitamins

Minerals are essential for production, reproduction and life of the animals. The

daily dietary requirement is dependent on the amount of dietary mineral that is absorbed

into the tissues (Goff, 2000). The requirement of the cow can be described by the dietary

requirement equal to maintenance plus pregnancy plus growth plus lactation divided by

the absorption co-efficient. In this equation, the upper portion can be obtained from NRC

(2001) or other table requirements. However the real challenge is to determine the co-

efficient of absorption for the minerals.

Coefficient of absorption for Ca was 0.38 in NRC (1989) and 0.45 in NRC (1978).

ARC (1980) determined a co-efficient of 0.68. Maybe a single coefficient is not

appropriate. Availability of Ca in forages, concentrates or mineral supplements is different

(Goff, 2000).

Coefficient of absorption for P was 0.50 in NRC (1989) and 0.65 in NRC (1978).

Rest of the world use 0.60 to 0.75.

The average coefficient of absorption for Mg from a wide variety of natural

feedstuffs fed to ruminants averaged 0.294 with a standard deviation of 0.135. The co-

efficient of absorption for Mg from inorganic sources should be 0.50 based on Mg oxide.

The coefficient for Mg absorption should be decreased when K is high in the diet (Goff,

2000)

Dietary Cl is absorbed with at least 80% and closer to 100% efficiency. Often Cl

anions accompany the movement of Na cations (Goff, 2000). Plants contain only small

amount of Na. Salt needs to be added to the diet, otherwise cows produce less milk.

Animals can tolerate very high levels of salt in the diet if water is provided and kidneys

are functioning. High dietary NaCI will reduce feed intake in animals (Goff, 2000).





83

Nearly all of the dietary K is absorbed. Intracellular uptake of K following a meal

helps buffer blood K concentration. The factorial approach as described for Ca, P and Mg

does not work for Na, K and Cl (Goff, 2000)

Rumen microbes need 0.2-0.22% S to operate efficiently. Excessive dietary S can

interfere with absorption of Cu and Se and may become toxic (Goff, 2000).

Cobalt is a component of vitamin B12. Microbes in rumen are the only natural

source of vitamin B12. Rumen microbes need 0.11% Co to perform efficiently (Goff,

2000).

Between 1 and 5% of dietary Cu will be absorbed by adult cattle. A diet high in Zn

(>1000 mg/kg), S and Mo can block Cu absorption. The Cu:Mo ratio should be 2:1 (Goff,

2000).

Selenium is part of glutathione peroxidase and in conjunction with vitamin E act

as antioxidant compounds. Selenium is also critical to thyroid hormone metabolism.

Selenium can cause deficiency or toxicity. Diets containing 0.1 ppm of Se are

recommended but field studies suggest this is not enough. Legally Se can be added up to

0.3 ppm (Goff, 2000).

Depending on the DMI the dietary requirement of iodine should be about 0.25-0.5

mg/kg DM (Goff, 2000).

Iron excess (250-500 mg Fe/kg DM) interfere with Cu and Zn absorption. NRC

(1989) recommends dietary Fe not to exceed 1000 mg/kg DM. This may be too high

(Goff, 2000).

Chromium is essential for normal glucose metabolism. The amount of Cr required

in the diet for optimal performance is unclear. More research is required (Goff, 2000).





84

The dietary coefficient of absorption for Zn is estimated to be 0.15. The major

dietary factors that can modify the efficiency of absorption of dietary Zn are interactions

of Zn with other metal ions (Cu, Fe) and the presence of organic chelating agents in the

diet (Goff, 2000).

The proportion of Mn absorbed from the diet generally is between 0.5 and 1%.

High dietary Ca, K or P increase Mn excretion in feces (Goff, 2000).

Mahanna (1997) recommends 0.3 ppm of Se dry matter basis. Potassium should

be not over 1.0% in the total ration for dry cows, but K Na and Mg should be 1.5, 0.5

and 0.35% of DM respectively in milking cows during heat stress. Nitrogen to sulfur ratio

should be 11 to 13:1 in the total ration to meet rumen bacterial needs. Goff and Horst

(1998a) recommend that dietary magnesium should be set at 0.4% (higher than NRC,

1989). Magnesium sulfate or magnesium chloride is recommended because they could be

an effective source of anions and magnesium.

Furthermore, the diet should supply between 35 to 50 g of phosphorus daily

(-0.4%). More than 80 g of P/day will inhibit renal synthesis of 1,25-dihydroxyvitamin D

which can induce MF. Dietary S should not exceed 0.4%. Adding more sulfate is a poor

choice because it is fairly ineffective acidifying agent. Dietary Cl can nearly always be

raised to 0.5% with little effect on dry matter intake. High dietary Ca concentrations (1.0-

1.2%) are desirable with anionic salts. Good results have been achieved by feeding as high

as 180 g Ca/day In one study, Joyce et al. (1997) determined that pre-calving cows fed

diets based on alfalfa (typically high in Ca) with a DCAD of- 7 had the lowest urine pH

prepartum and the highest concentrations of ionized Ca in blood. Also, this group

experienced the lowest incidence of calving related disorders. Results indicated that alfalfa

when supplemented with anionic salts is a viable forage for prepartum dairy cows.





85

Vitamin levels should be met to optimize milk production. Vitamin A should be

provided at rate of 3600 IU per kg DM/day. Vitamin D 900 IU per kg/DM day and

vitamin E 14 IU per kg/DM day (Mahanna, 1997).

Dry Cow Feed Management and Body Condition Score

Body condition scoring is a useful tool for monitoring the nutritional management

of dairy cows (energy density and intake). Using a scale 1 to 5 (Edmonson et al., 1989;

Ferguson et al., 1994) a program can be established. Beede (1997) recommends that, at

dry-off, cows should have a BCS 3.0 to 3.25. If BCS is lower, the ration should be

adjusted during the last 100 days of lactation and not during the dry period. If many cows

are over-conditioned, a fat cow group for late lactation should be established and a diet

containing 1.54 Mcal/Kg NE1/DM or less should be fed. If many cows are

underconditioned a thin cow group should be established and a diet with proper NEl

content to target the desired weight gain should be fed (one unit BCS s 57 kg BW).

Cows should have a total dry period length of at least 6 to 8 weeks. A dry period of

less than 6 weeks results in lower milk yields in the next lactation (Funk et al., 1987)

Two groups of dry cows are recommended; the early dry group (8 to 3 weeks

before expected calving) and a close-up group (3 weeks to calving). If early dry cows are

in proper condition and eating more than predicted to meet their requirements, it may be

necessary to lower the energy and nutrient density of the ration by re-formulating the diet.

Never try to reduce BCS or body weight of dry cows during any stage of the dry period (

Beede, 1997). If BCS is correct at dry-off (3.0-3.25) then cows should gain about 0.3-0.45

kg/d to increase BCS by 0.25 to 0.35 units during the early period.

In the close-up period, if DMI is around 11.3 kg or higher, a diet containing 1.6

Mcal/kg NE1 but no higher, and 14-15% of Crude Protein should be fed (NRC, 2001). A





86

separate close-up group for pregnant heifers might be a beneficial management strategy in

farms if adequate facilities are available (Grant and Albright, 1995). Diet formulation

should be based on 10 to 11 kg of DM intake. If cows are overeating it is not a big

problem because this is a short period of time. Cows should be in a positive energy status

and not losing weight. Feed should always be available in the bunk (24 h a day) in a form

of TMR to ensure adequate control of nutrient composition and consumption. Fermentable

grains (coarsely ground dry corn, hominy, or high moisture corn) to enhance development

of the ruminal epithelia should be fed (Beede, 1997). Cows should calve with a BCS of

3.5-3.75 and heifers 3.25-3.5 to minimize obstetrical trauma (Beede, 1997; Studer, 1998;

Grummer, 1999).

Fresh Cow Feed Management and Body Condition Score

The primary goal for early fresh cows is to maximize carbohydrate, protein and

nutrient intake and provide adequate fiber to meet requirements for increasing milk

production (Beede, 1997).

Mahanna (1997) recommends that cows should reach maximun daily DMI no later

than 10 weeks postpartum. Cows milked three times a day should eat about 5 to 6% more

DM per day. For every 1 kg of expected milk production cows should eat at least 0.45 kg

of DM. Eating less than this causes excessive body condition loss. Forage DM intake

should be near 2% of the cow's BW. Acid detergent fiber should be at least 19-21% and

NDF at least 28-30% or 0.9% of BW. Particle size should be long enough to stimulate 30

minutes of cud chewing time per kg of DM. Total mixed ration DM content should be

between 50 to 75%. Cows should reduce their DMI about 3.3% for every 1IC over 240C.

Heat stress starts when the temperature exceeds 26.7 C and relative humidity exceeds





87

80%. Clean water should be provided expecting cows to drink 2 liters for each 0.45 kg of

milk. Enough feed bunk space should be also provided.

Cows should peak in milk production 8 to 10 weeks after calving. First-calf heifers

should peak within 75% the production of older cows. For each extra 1 kg of milk at peak

production the average cow will produce 200 to 220 kg more milk for the entire lactation

period. Milk protein:fat ratio should be near 0.85-0.88 for Holstein cows. Forage intake

using a good quality roughage should be maximized.

Energy density of the top cows should be 1.76 Mcal/kg of EN1. Non fiber

carbohydrates levels should be between 35 and 42% with 25 and 35% starch in the total

ration.




Full Text
185
Paisley, L. G., W. D. Mickelson, and P. B. Anderson. 1986. Mechanisms and therapy for
retained fetal membranes and uterine intections of cows: A review. Theriogenology 25-353-
381.
Peeler, E. J., M. J. Otte, and R. J. Esslemont. 1994. Inter-relationships of periparturient
diseases in dairy cows. Vet. Rec. 134: 129-132.
Phipps, R.H., J.I.D. Wilkinson, L.J. Jonker, M. Tarrant, A.K. Jones, and A. Hodge. 2000.
Effect of monensin on milk production of Holstein-Friesian dairy cows. J. Dairy Sci. 83:2789-
2794.
Pimentel, S., S. G.Evans, W. C. Wagner. 1987. Placental synthesis of estrogens at parturition
and placental retention in the cow. Theriogenology 28:755-766.
Pintchuk P. A., F. D. Galey, L. W. George. 1993. Propylene toxicity in adult dairy cows. J.
Vet. Intern. Med. 7:150.
Plaizier, J. C.; A. Martin, T. Duffield, R. Bagg, P. Dick, and B. W. McBride. 2000. Effect of a
prepartum administration of monensin in a controlled-release capsule on apparent
digestibilities and nitrogen utilization in transition dairy cows. J. Dairy Sci. 83:2918-2925.
Prenge, R.W., C.L. Davis, and J.H. Clarke. 1978. Propionate production in the rumen of
Holstein steers fed either a control or monensin supplemented diet. J. Anim. Sci.
46:11201124.
Pugh, D. G., M. Q. Lowder, and J. G. Wenzel. 1994. Restrospective analysis of the
management of 78 cases of postpartum metritis in the cow. Theriogenology 42:455-463.
Radostits, O. M., K. E. Leslie, and J. Fetrow. 1994b. Culling and genetic improvement
programs for dairy herds. Pages 159-182. In Herd Health: Food Animal Production Medicine.
Second Edition. W. B. Sauders Company
Radostits, O. M., K. E. Leslie, and J. Fetrow. 1994c. Mastitis control in dairy herds.
Pages229-276. In Herd Health: Food Animal Production Medicine. Second Edition. W. B.
Sauders Company.
Rajala-Shultz, P. J., and Y. T. Grohn. 1998. Effects of dystocia, retained placenta and metritis
on milk yield in dairy cows. J. Dairy Sci. 81:3172-3181.
Rajala-Shultz, P. J., and Y. T. Grohn. 1999a. Culling of dairy cows. Part II. Effects of diseases
and reproductive performance on culling in Finnish Ayrshire cows. Prev. Vet. Med. 41:279-
294.
Rajala-Shultz, P. J., and Y. T. Grohn. 1999b. Culling of dairy cows. Part III. Effects of
diseases, pregnancy status and milk yield on culling in Finnish Ayrshire cows. Prev. Vet.
Med. 41:295-309.


167
Table 6-5 Summary of logistic regression modeling
Model
Dependent
Independent
OR1
Coefficients
95%CI2 P-value
Ketolactia
BHB milk
Monensin (yes)
0.68
0.53-0.80
0.014
> 200 pmol/L
Parity (primiparous)
1.05
0.85 1.34
0.39
Milk yield (< 30.4 1)
0.69
0.33 1.42
0.30
BCS (< 3.25)
0.63
0.32-1.26
0.19
1 Odds Ratio
2 95% Confidence Interval (Odd Ratio)


7
Reynolds, 1993). Similar to propionate, the contribution of these secondary glucose
precursors is partially dependent upon their supply and metabolic adaptation of transition
dairy cows (Drackley et al., 2001). Skeletal muscle, and to a lesser extent, skin, through
suppression of tissue protein synthesis and possibly increased proteolysis, serves as a
labile pool of amino acids that is mobilized to support increased gluconeogenesis during
the transition period (Bell, 1995; Bell et al., 2000). Alanine and glutamine account for 40
to 60% of the glucogenic potential of all the amino acids; therefore they typically make
the greatest contribution to glucose synthesis (Drackley et al., 2001). Alanine conversion
to glucose at 1 and 21 days postpartum was 198 and 150%, respectively, of that at 21 day
prepartum (Overton et al., 1998). Lactate utilization for gluconeogenesis primarily
represents recycling of carbons because most circulating lactate is formed either during
catabolism of glucose by peripheral tissues or by partial catabolism of propionate by
visceral peripheral tissues (Drackley et al., 2001). However, when non-forage fiber
sources and monensin were fed to transition prepartum cows, pyruvate carboxylase
expression was significantly induced at calving, suggesting an increased capacity of
peripartum cows for gluconeogenesis from lactate (Williams et al., 2003).
Non-esterified fatty acids (NEFA) concentrations are maximum at parturition
(0.9 to 1.2 mEq/L) with a slow decrease after 3 days postpartum (Melendez et al., 2002).
This finding corroborates the elevated fat mobilization occurring around parturition in
dairy cattle. Extreme rates of lipid mobilization lead to increased uptake of NEFA by liver
and increased triglyceride (TG) accumulation (Drackley, 1999). When blood glucose
concentrations increase lipogenesis predominates over lypolysis. This results in
suppression of NEFA release from adipose tissue (Herdt, 2000). The effect of glucose on
adipose tissue is related to insulin secretion and its role in glycerol synthesis, which is


43
(Kelton et al., 1998). Rajala-Shultz et al., (1999) reported in Finish Ayrshire cattle an
overall incidence of 3.3%, however, in some herds, ketosis can be a particular problem
and can affect a large proportion of at-risk cows (Herdt and Gerloff, 1999). Grohn et al.,
(1998) reported an overall incidence of 5.0% in New York Holstein cattle. They
determined a range between 4 to 22% among 8 herds as well (Grohn et al. (1999).
Gstergaard and Grohn, (1999), in Danish cattle determined an incidence of ketosis of
2.0% in primiparous cows and 10.0% in multiparous cows. Al-Rawashdeh, (1999) in
Jordanian dairy cattle determined an overall prevalence of mild ketonemia (0.9 1.7
mmol/1 of BHB) of 22% and severe ketonemia (> 1.7 mmol/1 of BHB) of 3.8%.
More than 90% of subclinical ketosis cases occur in the first and second months
after calving. During this period, approximately 40% of all cows are affected by
subclinical ketosis at least once, although the incidence and prevalence are highest in the
first and second weeks after parturition (Duffield, 2000; Geishauser et al., 2001). In a
Finish study, the odds of contracting ketosis during the first 39 days in milk was higher
than cows beyond 39 days postpartum (Schnier et al., 2002).
Economic losses from ketosis include treatment of clinically ill cows, lost milk
production, increased days open, and increased culling, and were estimated to total
approximately $145 per case (Guard, 1994). Seven studies found short- or long- term
reduction in milk yield associated with clinical ketosis, whereas four reported no losses
(Fourichon et al., 1999). Five studies found losses associated with ketotic status according
to a diagnostic test, whereas two reported no losses (Fourichon et al., 1999). These
discrepancies might be explained by the different statistical methods used to analyze the
effect of the disease on milk production. Rajala-Shultz et al. (1999), using repeated
measures, based on a mixed model with a special parametric structure for the covariance


164
Table 6-2. Nutrient content of far-off dry cow, transition dry cows and lactation
transition diets
Nutrient
Dry cow
far-off
Dry cow
transition
Lactating
transition
CP(%DM) '
15.52
17.91
18.60
UndegP(%CP) 2
26.38
35.31
30.34
DegP(%CP)2
73.62
64.69
69.66
SolP(%CP)2
-
36.72
39.77
NEL(Mcal/kg)3
0.84
1.69
1.69
ADF(%DM) 1
24.54
25.13
23.66
NDF(%DM) 1
36.04
36.20
34.63
NFC(%DM) 2
32.94
31.55
34.13
Starch(%DM) 2
11.05
14.96
14.90
Lipid (%DM) 1
2.85
6.20
2.36
Ca(%DM)1
0.74
1.27
1.10
P(%DM) 1
0.30
0.35
0.46
Mg(%DM) 1
0.32
0.36
0.36
K(%DM) 1
1.28
1.10
1.46
Na(%DM) 1
0.09
0.09
0.58
C1(%DM) 1
0.18
0.43
0.48
S(%DM) 1
0.24
0.40
0.22
Forage in diet (%DM)
60.21
48.10
50.23
Cation-Anion (meq/kg DM) 4
165.90
-51.70
351.90
1 Laboratory nutritional analysis
2 Values from feed composition tables
3 From formulas after laboratory analysis
4 From formula (Na+ + K+) (CF + S')


132
Table 4-6. Least squares means S.E.M. for serum B-hydroxy butyrate, non-esterified fatty
acids and glucose by time of sampling after A.M. feeding in multiparous treated and
control cows at 10 d pp
Time
Group
BHB
mmol/L
NEFA
meq/L
Glucose
mg/dL
TO
M
0.83 0.10
1.01 0.08
48.9 2.20
C
1.01 0.13
1.04 0.11
46.9 3.10
T2
M
0.83 0.10
0.59 0.08f
51.2 2.20
C
0.96 0.13
0.92 0.11t
46.3 3.10
T4
M
0.770.10n
0.60 0.08
48.6 2.20
C
1.05 0.13+t
0.71 0.11
47.3 3.10
T6
M
0.820.10t
0.60 0.08
47.4 2.20
TFTTTTr
C
1.18 0.13+
0.60 0.11
48.6 3.10
' (P <0.1)
+ (p < 0.05)
TO: at A.M. feeding; T2: 2 h post feeding; T4: 4 h post feeding; T6: 6 h post feeding


186
Rajala-Shultz, P. J., Y. T. Grhn, and C. E. McCulloch. 1999. Effects of milk fever, ketosis
and lameness on milk yield in dairy cows. J. Dairy Sci. 82:288-294.
Ramanzin, M., L. Bailoni, S. Schiavon, and G. Bittante. 1997. Effect of monensin on milk
production and efficiency of dairy cows fed two diets differing in forage to concentrate raios
J. Dairy Sci. 80:1136-1142.
Reist, M., Erdin, D., von Euw, D, Tschmperlin, K., Leuenberger, H., Hammon, H., Knzi,
N., and Blum, J. 2003. Use of threshold serum and milk ketone concentrations to identify risk
for ketosis and endometritis in high-yielding dairy cows. Am. J. Vet. Res. 64:188-194.
Reynolds, C.K., P.C. Aikman, D.J. Humphries, and D.E. Beever. 2000. Splanchnic
metabolism in transition dairy cows. J. Dairy Sci. 83(Suppl. 1): 257. (Abstr.)
Richardson, L.F., A.P. Raun, E.L. Potter, C.O. Cooley, and R.P. Rathmacher. 1976. Effect of
monensin on rumen fermentation in vitro and in vivo. J. Anim. Sci. 43:657-664.
Risco, C. A., L. F. Archbald, J. Elliott, T. Tran, and P. Chavatte. 1994a. Effect of hormonal
treatment on fertility in dairy cows with dystocia or retained fetal membranes at parturition. J.
Dairy Sci. 77:2562-2569.
Risco, C. A., M. Drost, W. W. Thatcher, J. Savio, M. J. Thatcher. 1994b. Effects of calving-
related disorders on prostaglandin, calcium, ovarian activity and uterine involution in
postpartum dairy cows. Theriogenology 42:183-203.
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monensin on the performance and nitrogen utilization of lactating dairy cows consuming fresh
forage. J. Dairy Sci. 84:1717-1727.
Rukkwamsuk, T., M.J.H. Geelen, T.A.M. Kruip, and T. Wensing. 2000. Interrelation of fatty
acid composition in adipose tissue, serum, and liver of dairy cows during the development of
fatty liver postpartum. J. Dairy Sci. 83:52-59.


70
Monensin primarily has been used as a powder mixed with concentrates.
Recently a capsule of slow release of monensin has been developed. In Australia, Lean at
al. (1994) and Abe et al. (1994) conducted two studies using a slow-release intraruminal
capsule containing 32 g of sodium monensin. Since that time, capsules have been
available in Australia and Canada.
Lean at al. (1994) postulated that feeding sodium monensin may result in
additional glucose flux and reduced risk of reproductive failure associated with
hypoglycemia and that monensin feeding would increase milk and milk solid yields. They
used a total of 1061 lactating dairy cows from six different herds and management. One
herd was fed a total mixed ration, three were fed on pasture and supplemented
concentrates and two herds were primarily fed on pasture. Capsule was administered
within 7 days of calving. Conception rate at first service, days to first estrus and calving-
to-conception interval did not differ significantly between untreated and monensin-treated
cows. Monensin treatment significantly increased milk yield only in one herd. Milk fat
and protein were not significantly affected by treatement. Abe et al. (1994) wanted to
evaluate the effects of a sodium monensin controlled-release capsule on metabolic
responses and on resumption of ovarian cyclicity. Cows treated with monensin had
significantly lower plasma BHB and tended to have higher plasma glucose than untreated
cows. Treatment did not significantly influence plasma NEFA, urea nitrogen or
cholesterol concentrations. Time to first estrus was significant shorter for untreated than
for treated cows, however inclusion of BCS immediately after calving as a covariate
resulted in non-significant differences. Time to first ovulation did not significantly differ
between groups.


LIST OF TABLES
Table page
2-1. Predicted changes in dry matter intake for Holstein dairy
cows during the last three weeks prior to calving 5
2-2. Effect of hormones on carbohydrates and lipids metabolites
on dairy cattle 10
2-3. Summary of relationships among Calving-Related Disorders 11
2-4. Case definition, incidence and economic losses of
calving-related disorders 14
2-5. Ketone body concentrations for clinical and subclinical
ketosis according to different authors 56
2-6. Ketone body field test comparison 57
2-7. Relative volumes and number of microbial organisms 60
2-8. Susceptibility and resistance of ruminal bacteria to monensin 67
2-9. Minimum requirements for dry, prepartum and fresh cows 88
2-10. Minimum trace mineral and vitamins requirements for dry,
prepartum and fresh cows 89
2-11. Target Body Condition Scores (BCS) Scale 1-5 90
3-1. Diet composition dry cow far-off, dry cow transition and lactating
Transition cows 105
3-2. Nutrient content dry cow far-off, dry-cow transition and lactating
transition diets 106
3-3. Lactational Incidence of Calving-Related Disorders 107
3-4. Summary of Logistic Regression Modeling for CRD 108
VII


6
and for cows is equal to 1.97 0.75 e016t, where t is equal to days of pregnancy minus
280.
Glucose and Lipid Metabolism
Glucose and amino acids are the major fuel supply of the developing fetus in
ruminants. Glucose and amino acids are also needed by the mammary gland for lactose
and milk protein synthesis, respectively (Herdt, 2000). Glucose demand in Holstein cows
has been estimated at 1000 to 1100 g/d during the last 21 d of gestation, but increases
sharply after calving to approximately 2500 g/d at 21 d postpartum (Drackley et al., 2001).
Ruminants are not entirely dependent on dietary glucose; as a result they are in a constant
stage of gluconeogenesis (Herdt, 2002). The liver serves as a linchpin in adaptation to the
maintenance of body fuel supplies and consequently it is the key regulator of glucose
supply to the tissues (Herdt, 2000). The major gluconeogenic precursor in ruminants is
propionic acid produced in the rumen. Its contribution to gluconeogenesis has been
estimated to be 32 to 73% (Seal and Reynolds, 1993). Liver uptake of propionate by portal
circulation is almost 100% (Herdt, 2002); however the capacity of the liver to convert
propionate to glucose seems to be responsive to the amount of propionate supplied and the
physiological stage of the animal (Drackley et ah, 2001). Hepatic propionate metabolism
is modulated during the transition period. As an example, hepatic blood flow in cows
increases 84% from lid prepartum to 11 d postpartum (Reynolds et ah, 2000). In
addition, propionate conversion to glucose by the liver is 19 and 29% greater at day 1 and
21 postpartum, respectively, than at day 21 prepartum (Overton et ah, 1998).
Amino acids, lactate and glycerol are secondary substrates for gluconeogenesis in
ruminants (Herdt, 2002). Contribution to glucose production has been estimated to be 10
to 30% for amino acids, 15% for lactate and small amounts for glycerol (Seal and



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3 STUDY 1 91
Introduction 91
Material and Methods 92
Results 97
Discussion 98
4 STUDY 2 118
Introduction 118
Material and Methods 119
Results and Discussion 123
5 STUDY 3 140
Introduction 140
Material and Methods 141
Results and Discussion 144
6 STUDY 4 155
Introduction 155
Material and Methods 156
Results and Discussion 160
REFERENCE LIST 169
BIBLIOGRAPHICAL SKETCH 192
VI


144
were considered significant at P < 0.05. Statistical analysis was conducted using the Proc Mixed
procedure of SAS 8.2 (SAS, 2001).
Mixed models for repeated measures were defined as:
yijkim= B + Ti + Timej + Park +BCS, + Dysm + (Time*T)ij + (T*BCS)¡, +
(T*Dys)jm + (Par *Time*T)ijk + eijklrn
Where:
y¡jkim= Blood or rumen metabolites
Tj= fixed effect of treatment
Cow (T¡)j = random effect of cow nested in treatment
Time k = fixed effect of time
Par i = fixed effect of parity
BCS m = random effect of Body Condition Score at Calving
Dysm = random effect of dystocia
(Time *T) ¡k = interaction time and treatment
(T*BCS)n = interaction treatment and body condition score at calving
(T*Dys)¡m = interaction treatment and dystocia
(Par *Time*T) k¡i = interaction time, parity and treatment
eijkim random error term
ANOVA Mixed models were defined as:
yijk= p + Tj + Parj + (Par*T)j¡ + eijk
Where:
y¡jk= BCS at calving or accumulated real and ME 305 milk production
T¡= fixed effect of treatment
Cow (Tj)j = random effect of cow nested in treatment
Parj = fixed effect of parity
(Par *T) ¡j = interaction parity and treatment
e¡jk = random error term
Results and Discussion
Body Condition Score
In Figure 5.1, BCS at assignment and at calving, by treatment and by parity are reported.
Body condition score at assignment and milk yield of previous lactation were similar between
groups within parity (P > 0.05). There was no treatment effect on BCS at calving in primiparous


172
Correa, M. T., C. R. Curtis, H. N. Erb, J. M. Scarlett, and R. D. Smith. 1990. An ecological
analysis of risk factors for postpartum disorders of Holstein-Friesian cows from thirty-two
New York farms. J. Dairy Sci. 73:1515-1524.
Correa, M. T., H. Erb, and J. Scarlett. 1993. Path analysis for seven postpartum disorders of
Holstein cows. J. Dairy Sci. 76:1305-1312.
Cox, D. R. 1972. Regression models and life tables (with discussion). J. of Royal Statistical
Soc.: Series B, 34:187-220.
Crawley, D. D., and L. H. Kilmer. 1995. Effect of level and source of rumen degradable
protein fed prepartum on postpartum performance of dairy cows. J. Dairy Sci. 78(Suppl.
1):266.
Curtis, C. R., H. N. Erb, C. J. Sniffen, and R. D. Smith. 1984. Epidemiology of parturient
paresis: predisposing factors with emphasis on dry cow feeding and management. J. Dairy
Sci. 67:817-825.
Curtis, C. R., H. N. Erb, C. J. Sniffen, R. D. Smith, and D. S. Kronfeld. 1985. Path analysis of
dry period nutrition, postpartum metabolic and reproductive disorders, and mastitis in
Holstein cows. J Dairy Sci. 68:2347-2360
Curtis, C. R.; H. N. Erb, C. J. Sniffen, R. D. Smith, P. A. Powers, M. C. Smith, M. E. White,,
R. B. Hillman, and E. J. Pearson. 1983. Association of parturient hypocalcemia with eight
periparturient disorders in Holstein cows. J. Amer. Vet. Med. Assoc. 183:559-561.
Dawson, L. J., E. P. Aalseth, L. E. Rice, and G. D. Adams. 1992. Influence of fiber form in a
completed mixed ration on incidence of left displaced abomasum in postpartum dairy cows. J.
Amer. Vet. Med. Assoc. 200:1989-1992.
De Boer, G., A. Trenkle, and J. W. Young. 1985. Glucagon, insulin, growth hormone, and
some blood metabolites during energy restriction ketonemia of lactating cows. J. Dairy Sci.
68:326-337.
Dechow, C.D.; G.W. Rogers, and J.S. Clay. 2001. Heritabilities and correlations among body
condition scores, production traits, and reproductive performance. J. Dairy Sci. 84:266-275.
DeGraves, F. J. and J. Fetrow. 1993. Economic of mastitis and mastitis control. Vet. Clinics
North Amer. 9: 421-434.
Delaquis, A. M., and E. Block. 1995. Dietary cation-anion difference, acid-base status,
mineral metabolism, renal function, and milk production of lactating cows. J. Dairy Sci.
78:2259-2284.
Deluyker, H. A., J. M. Gay, L. D. Weaver, and A. S. Azari. 1991. Change of milk yield with
clinical diseases for a high producing dairy herd. J. Dairy Sci. 74:436-445.


55
oxaloacetate and finally to glucose by the liver (Casse et al., 1994). In Figure 2-1 major
energy biochemical pathways in cattle are summarized.
Nutritionally, propionate can be supplied orally in the form of sodium propionate
or calcium propionate. Calcium propionate has been studied as a calcium-energy substrate
for dairy cows (Melendez et al., 2002a). Calcium propionate is a gluconeogenic precursor
as well as a source of calcium, which will be absorbed by the small intestine and may help
to prevent clinical and subclinical hypocalcemia (Goff et al., 1996). Calcium propionate is
a compound poorly fermented by the rumen microorganism. At parturition, calcium
propionate increases blood glucose 24 hrs after its administration, and reduces BHB and
NEFA during the first two days postpartum. Furthermore, calcium propionate increases
blood calcium, reduces the incidence of clinical and subclinical hypocalcemia and
increases milk yield by 3.8 kg/d during the first 2 weeks after calving (Higgins et al.,
1996). In transition cows fed anionic salts prepartum, a calcium propionate (510 g) plus
propylene glycol (400 g) drench did not affect postpartum concentrations of Ca, P, Mg,
glucose, NEFA, or BHB (Melendez et al., 2002a).


60
hydrogen producing-bacteria such as Ruminococcus and Butyrivibrio and thereby favoring
propionate producers (Van Soest, 1994).
Rumen Microbial Ecosystem
The adult rumen evolved to aid the digestion of grasses. The rumen microbes
hydrolyze the plant celluloses, hemicelluloses, pectins, ffuctosans, starches and other
polysaccharides to monomeric or dimeric sugars which are further fermented to give
various products such as acetic, propionic, and butyric acids, methane and carbon dioxide
(Hobson, 1997).
Rumen microorganisms are predominantly strict anaerobes. Some oxygen can be
tolerated as long as the fermentation is sufficiently active to facilitate the disposal of
oxygen and the potential (Eh) of the medium remains within normal limits (-250 to -450
mV). Oxygen introduced through feed and water may diffuse across the rumen wall and
affect organisms near the wall, where oxygen serves as an electron acceptor. Facultative
anaerobes consume oxygen and help to maintain the low Eh (Van Soest, 1994).
Rumen microorganisms are represented by bacteria, protozoa and fungi. In Table
2-7, relative volume and number of microbial organisms are shown.
Table.2-7. Relative volumes and number of microbial organisms
Group
Number
mL
Mean cell
volume (p3)
Net mass
(mg/dL)
Generation
time
% of total rumen
microbial mass
Small bacteria
1 x 10IU
1
1600
20 min
60-90
Selenomonads
1 x 108
30
300
-
-
Large bacteria
1 x 106
250
25
-
-
Protozoa
3x 109
1 x 109
300
8 h
10-40
Fungi
1 x 104
1 x 105
-
24 h
5-10
Adapted from: Van Soest, P.J. 1994.Microbes in the gut. Pages 253-280 in Nutritional
ecology of the ruminant. Second edition. P. J. Van Soest. Cornell University Press.


177
Greenfield, R.B., M.J. Cecava, and S.S. Donkin. 2000. Changes in mRNA expression for
gluconeogenic enzymes in liver of dairy cattle during the transition to lactation J Dairy Sci
83:1228-1236.
Greenough, P. R., A. D. Weaver, D. M. Broom, R. J. Esslemont, and F. A. Galindo. 1997.
Basic concepts of bovine lameness. Pages 3-13. In Lameness in cattle. P. R. Greenough and
A. D. Weaver. Third Edition. W.B. Saunders Company.
Greiner, M., D. Pfeiffer, and R.D. Smith. 2000. Principles and practical application of the
receiver-operating characteristic analysis for diagnostic tests. Prev. Vet. Med. 45:23-41.
Grohn Y. T., S. W. Eicker, V. Ducrocq, and J. A. Herd. 1998. Effect of diseases on the
culling of Holstein dairy cows in New York state. J. Dairy Sci. 81:966-978.
Grhn Y. T., H. N. Erb, C. E. McCulloch, and H. S. Saloniemi. 1989. Epidemiology of
metabolic disorders in dairy cattle: Association among host characteristics, disease, and
production. J. Dairy Sci. 72:1876-1885.
Grohn Y. T., H. N. Erb, C. E. McCulloch, and H. S. Saloniemi. 1990a. Epidemiology of
reproductive disorders in dairy cattle: associations among host characteristics, disease and
production. Prev. Vet. Med. 8:25-39.
Grohn Y. T., J. J. McDermott, Y. H. Schukken, J. A. Herd, and S. W. Eicker. 1999.
Analysis of correlated continuous repeated observations: modelling the effect of ketosis on
milk yield in dairy cows. Prev. Vet. Med. 39:137-153.
Gram, D. E., J. K. Drackley, R. S. Younker, D. W. LaCount, and J. J. Veenhuizen. 1996.
Nutrition during the dry period and hepatic lipid metabolism of periparturient dairy cows. J.
Dairy Sci. 79:1850-1864.
Grammer, R. R. 1993. Etiology of lipid-related metabolic disorders in periparturient dairy
cows. J. Dairy Sci. 76:3882-3896.
Grummer, R. R. 1995. Impact of changes in organic nutrient metabolism on feeding the
transition dairy cows. J. Anim. Sci. 73:2820-2833.
Grammer, R. R. 1999. Energy and protein nutrition of the transition dairy cow. In Dry cow
nutrition. 35th Annual Conference of the American Association of Bovine Practitioners.
Nashville, TN.
Grammer R. R., J. C. Winkler, S. J. Bertics, and V. A. Studer. 1994. Effect of propylene
glycol dosage during feed restriction on metabolites in blood of prepartum Holstein heifers. J.
Dairy Sci. 77:3618-3623.
Granert, E. 1986. Etiology and pathogenesis of retained placenta. Pages 237-In Current
Therapy in Theriogenology 2. D. A. Morrow .Philadelphia, WB Saunders.


37
Mastitis
Mastitis is the most common and costly disease of dairy cows. The major
economic loss of all forms of mastitis results from reduced milk production (De Graves
and Fetrow, 1993). Mastitis losses have been estimated at $2 billion per year in the U.S.A
or approximately $200 per cow annually (Fetrow and Anderson, 1987).
Mastitis may be clinical or subclinical. Clinical mastitis is defined as visually
abnormal milk secretion (e.g., clots, flakes, or watery) from one or more quarters. It might
or might not be accompanied by signs of inflammation of the udder tissue (e.g., heat,
swelling, or discoloration of the skin (Kelton et al., 1998). Subclinical mastitis is defined
as the presence of pathogenic microorganisms in the milk and a somatic cell count (SCC)
above 500,000 cells/ml. Reduced milk production associated with subclinical mastitis
accounts for approximately 70% of the economic loss. Treatment costs, culling, and
reduced productivity associated with clinical mastitis are responsible for the remaining
losses (Radostits et al., 1994). There is a straight line negative relationship between a
logarithmic transformation of SCC (LS) and milk production. Estimates of milk
production losses range from 3 to 6% with each unit increase of the LS (Barlett et al.,
1990). Losses associated with the treatment of clinical mastitis have been estimated at
more than $100 per episode (Hoblet et al., 1991).
The reported frequency of clinical mastitis, based on 62 citations from 1982 to
1996 ranged from a lactational incidence of 1.7% to 54.6%. A new case was defined when
abnormal milk in a different or the same quarter followed at least 8 days of normal milk
(Kelton et al., 1998).
The causes of bovine mastitis have been classified as major pathogens and minor
pathogens. The major mastitis pathogens can be further classified as causes of contagious


189
Studer, E. 1998. A veterinary perspective of on-farm evaluation of nutrition and reproduction
J. Dairy Sci. 81:872-876.
Studer V. A., R. R. Grummer, S. J. Bertics, and C. K. Reynolds. 1993. Effect of prepartum
propylene glycol administration on periparturient fatty liver in dairy cows. J. Dairy Sci
76:2931-2939.
Trent, A. M. 1990. Surgery of the bovine abomasum. Vet. Clin. North Am. Food Anim. Pract
6:399-448.
United States Department of Agriculture Animal and Plant Health Inspection Service -
Veterinary Services. 1997. Papillomatous digital dermatitis on U.S. dairy operations. NAHMS
Dairy 96. USDA-APHIS-VS, Ft. Collins, CO. 28 p.
Vagnoni, D. B., and G. R. Oetzel. 1998. Effects of dietary cation-anion difference on the acid-
base status of dry cows. J. Dairy Sci. 81:1643-1652.
Vallimont, J.E., G.A. Varga, A. Arieli, T.W. Cassidy, and K.A. Cummins. 2001. Effects of
prepartum somatotropin and monensin on metabolism and production of periparturient
Holstein dairy cows. J. Dairy Sci. 84:2607-2621.
VandeHaar, M. J., B. K. Sharma, G. Yousif, T. H. Herdt, R. S. Emery, M. S. Allen, and J. S.
Liesman. 1995. Prepartum diets more nutrient-dense than recommended by NRC improve
nutritional status of peripartum cows. J. Dairy Sci. 78(Suppl. 1):264.
VandeHaar, M. J., G. Yousif, B. K. Sharma,, T. H. Herdt, R. S. Emery, M. S. Allen, and J. S.
Liesman. 1999.Effect of energy and protein density of prepartum diets on fat and protein
metabolism of dairy cattle in the periparturient period. J. Dairy Sci. 82:1282-1295.
Van der Werf, J. H. J., L. J. Jonker, and J. K. Oldenbroek. 1998. Effect of monensin on milk
production by Holstein and Jersey cows. J. Dairy Sci. 81:427-433.
Van Dorp, T. E., J. C. M. Dekkers, S. W. Martin, and J. P. Noordhuizen. 1998. Genetic
parameters of health disorders, and relationships with 305-day milk yield and conformation
traits of registered Holstein cows. J. Dairy Sci. 81:2264-2270.
Van Maanen, R.W., J.H. Herbein, A.D. McGilliard, and J.W. Young. 1978. Effects of
monensin on in vivo rumen propionate production and blood glucose kinetics in cattle. J.
Nutr. 108:1002-1007.
Van Soest, P.J. 1994.Microbes in the gut. Pages 253-280 in Nutritional ecology of the
ruminant. Second edition. P. J. Van Soest. Cornell University Press.
Van Werven, T., Y. H. Schukken, J. Lloyd, A. Brand, H. Heeringa, and M. Shea. 1992. The
effect of duration of retained placenta on reproduction, milk production, postpartum disease
and culling rate. Theriogenology 37:1191-1203.


NEFA (mEq/L) Glucose (mmol / L) BHB (mmol/L)
137
2.5
2.25
2
Control
Figure 4-3. Effect of monensin treatment on glucose, BHB, and NEFA concentration in
multiparous cows as a function of time after feeding at 10 d pp


101
(Duffield et al., 2002). In addition, the transition diet supplied 1.67 Mcal/kg DM of NE1
and glucose precursors such as milk whey, sodium propionate and propylene glycol
(Metaxerol and Lactowhey) (see Table 3.1) and perhaps this diet was sufficient to
keep an adequate energy balance, normal glycemia and low incidence of clinical ketosis
in transition cows without monensin.
The incidence of DA was within the normal ranges of 0.3 to 6.3% reported in the
literature (Kelton et al., 1998). There was no effect of treatment even though the control
group had an incidence 2 times higher than treated groups. The sample size relative to
incidence of DA may have reduced the power to detect differences due to treatments
(power=14% or 1433 animals per group). Similar results were reported in one of the
Canadian studies. They found an incidence of DA almost double in the control vs treated
group (5.3% vs 3.0%, respectively) with a P < 0.20 (Duffield et al., 1999b).
Culling rate was similar between groups (28.6% respectively) (P > 0.05). Similar
conclusion was reported in a Canadian study (Duffield, 2000).0verall, culling rate is
within the normal values reported in the scientific literature (Radostits, 1994). Milk
fever, RFM, displacement of abomasum, ketosis and ovarian cysts significantly affect
culling at different stages of lactation. Metritis has no effect on culling (Radostits, 1994;
Grohn et al., 1998). Since the incidence of milk fever, RFM, DA and clinical ketosis was
similar between treated and control animals it is reasonable the fact that culling rate was
similar between groups.
Body Condition Score
An interesting finding of the present study was that monensin given at dry-off
improved slightly BCS at calving in older cows. Within parity, control and treated cows
had similar BCS at dry-off (P>0.05). However the difference was significant for


44
matrices, found a significant negative effect of ketosis on milk production. The milk-
reducing effect started even before the diagnosis of clinical ketosis. Milk losses continued
for at least 2 weeks after diagnosis and the overall losses during the entire lactation ranged
from 126 kg in parity 1 to 535 kg in parity 4 and an average total loss per cow of 353 kg.
Daily milk loss within the first two weeks after diagnosis was 3.0, 4.0, 3.3, and 5.3 kg/d
for parities 1, 2, 3, and 4 or higher, respectively. In parity 4 or higher, the milk loss
continued for the rest of the lactation. This could be an indication that the energy
requirements of the cow were not met. However, healthy cows overall produced less milk
than ketotic cows in the same parity. This might indicate that cows with ketosis are, in
general, higher yielding and that the milk loss often is only temporary. The same
conclusion was observed in New York Holstein cattle when they did a modeling of the
effect of ketosis on milk yield, using a mixed model with four different covariance
structures Grohn et al., (1999). By using projected 305-day milk yield, ketosis had no
effect on milk production; but by using a monthly test-day milk as a repeated
measurements and four covariance structures, ketotic cows yielded significantly less milk
per day both before and immediately after diagnosis than did non-ketotic cows. In another
study, 0stergaard and Grohn (1999), in Danish cattle, found that ketotic cows had
significantly higher milk yield before the disease that did healthy cows. Multiparous cows
lost between 1.5 and 4.2 kg/day of milk between 1 to 4 weeks after diagnosis of ketosis.
Furthermore, the body weight of cows with ketosis was lower after diagnosis than was that
of healthy cows. In Swiss cattle, a significant decrease in milk production (442 to 654 kg
of energy-corrected milk/305-day period per cow) was associated with acetone or BHB in
excess of threshold values of > 0.40 mmol/L and > 2.3 mmol/L, respectively (Reist et al.,
2003).


124
monensin treatment is decreased by more than 30% and this decrease corresponded to about a
10-fold decrease in the numbers of bacteria that ferment peptides and amino acids as an energy
source for growth (Yang and Russell, 1993). Ionophores have also been demonstrated to
decrease ruminal urease activity (Starnes et al., 1984). Other authors did not find positive
effects of monensin on ammonia production; however their sample size per groups was only 8
cows or less per group (Ramanzin et al., 1997: Plaizeir et al., 2000).
Blood Metabolites
At assignment, cows did not differ in BCS, days BEP, parity and previous lactation
mature equivalent milk production. Body condition score changed over time, with an
interaction of treatment by parity. Treatment affected BCS significantly only in multiparous
cows (Figure 4.1) (P < 0.01). Perhaps monensin altered the tissue composition of the daily gain
in adult cows, favoring the deposition of fat instead of protein. Indeed, daily gains in adult
cattle contain more fat, less protein, and less water than daily gain in growing cattle (Williams
et al., 1989).
In Table 4.5 and 4.6 least squares means and SEM for blood metabolite concentrations
at 10 d pp for primiparous and multiparous cows in treated and control groups are shown. In
Figures 4.2 and 4.3 blood metabolite concentrations patterns at 10 d pp for primiparous and
multiparous cows in treated and control groups are illustrated. In Tables 4.7 and 4.8 least
squares means and SEM for blood metabolites concentrations between dry-off and 21 d pp for
primiparous and multiparous cows in treated and control groups are shown. In Figures 4.4 and
4.5 blood metabolite concentrations patterns between dry-off and 21 d pp for primiparous and
multiparous cows in treated and control groups are illustrated.


76
rumen butyrate. The rumen concentrations of acetate and propionate were not different.
Mean serum BHB concentration did not differ between groups during the 2 week
postpartum period, although cows treated with monensin tended (P < 0.06) to have a lower
concentration than controls. During the 6 week postpartum period cows treated with
monensin had significantly lower BHB concentration than controls. Control cows had a
serum BHB concentration that exceeded 1200 pmol/L by week throughout the postpartum
period. They concluded that monensin maintained energy-related blood metabolites,
reduced the occurrence of subclinical ketosis, and improved rumen health in the early
lactation dairy cow.
The effect of a monensin controlled-release capsule on diet digestibility and
nitrogen utilization was investigated in 16 multiparous dairy cows between approximately
10 and 3 days precalving and 3 and 9 days postpartum. Monensin decreased significantly
fecal nitrogen output and improved nitrogen balance. The control group lost on average
77.8 g/day of nitrogen, compared with 44.9 g/day in the treated group. The positive effect
of monensin on NDF and ADF digestibility precalving and nitrogen digestibility
postcalving was also significant (Plaizier et al., 2000).
The same Canadian group, studied the effects of monensin administered either as
a controlled release capsule or as a premix, on attenuating grain-induced subacute ruminal
acidosis and on ruminal fermentation characteristics in Holstein cows receiving a total
mixed ration (Mutsvangwa et al., 2002). They used six multiparous Holstein cows in a
two-treatment, two-period crossover design with 6 week periods. Experiment 1 used a
monensin bolus and experiment 2 used a monensin premix. Authors induced subacute
rumen acidosis between day 22 and 32 of each experimental period, by restricting feed
intake to 85% and substituting 15% of the diet with grain pellets of wheat and barley.


15
Milk Fever. Parturient Paresis. Hypocalcemia
Milk fever (MF), is a non-febrile metabolic disease affecting milking cows in
which acute calcium deficiency causes progressive neuromuscular dysfunction with
flaccid paralysis, circulatory collapse, and depression of consciousness (Oetzel and Goff,
1999).
The reported frequency of MF, based on 33 citations from 1979 to 1995 ranged
from 0.03% to 22.3%. The median lactational incidence was 6.5% (Kelton et al., 1998).
However, Oetzel and Goff (1999) established that annual incidence rate within herds may
vary from 2% to 60%. The incidence of MF reported in a Florida commercial dairy herd
using anionic diets during the prepartum period was 0.42% (2/477) (Melendez et al.,
2003b). Approximately 75% of all cases of MF occur within 24 hours of calving. An
additional 12% occur 24 to 48 hours after calving. Some cases (about 6%) occur at the
time of delivery and cause dystocia because hypocalcemia induce uterine atony (Oetzel
and Goff, 1999). Only 3% of cases occur prepartum, and only 4% occur more than 48
hours postpartum (Oetzel, 1988).
Hypocalcemia may be clinical or subclinical. Clinical signs of MF are not seen
until calcium is about 4 mg/100 ml (Goff and Horst, 1997b). Subclinical hypocalcemia
affects about 50% of all adult dairy cattle. In this case, plasma Ca concentration of
periparturient cows remained < 7.5 mg/dL, even up to 10 days after calving (Goff et al.,
1996). This condition may lead to decreased dry matter intake after calving, increased risk
of secondary diseases, decreased milk production and decreased fertility (Goff and Horst,
1997b).
Economic losses from MF include prophylactic or clinical treatment, lost milk
production, poor reproductive performance, and culling, which are estimated to total


102
multiparous cows at calving (P < 0.05). The effect of monensin as a growth promotant in
females has been described in beef and dairy cattle (Baile et al., 1982; Simpson et ah,
1998). In dairy heifers, monensin did not change body composition (Meinert et ah,
1992), but in adult cows monensin increased either body weight, BCS or reduced BCS
losses after calving (Duffield et ah, 1998a; Van Der Werf et ah, 1998; Phipps et ah,
2000), which is in agreement with the present study. Monensin did not increase BCS at
calving in first lactation cows (P > 0.05), but did increase it in second lactation and older
cows (P < 0.05). This improvement did not, however, put cows in a detrimental obese
condition.
Milk Yield and Milk Solids
In the present study, monensin increased milk yield slightly. These findings have
been consistent with previous studies using monensin as a powder or as a slow-release
bolus (Lean et ah, 1994; Hayes et ah, 1996; Van Der Werf et ah, 1998; Duffield et ah,
1999a; Phipps et ah, 2000; Mutsvangwa et ah, 2002). Other studies did not find a
positive effect of monensin on milk production (Abe et ah, 1994; Green et ah, 1999;
Vallimont et ah, 2001). Monensin increased milk yield during the entire lactation, based
on test day milk yield, in third lactation or older animals, but not in first lactation cows.
The positive effect of treatment in the short and long term might be explained by two
different physiologic mechanisms of the ionophore. The bolus lasts for 95 days,
therefore the short term positive response might be related to a direct effect of monensin
during the first 20 d of lactation, which is concomitant with the last 20 to 30 days of the
duration of the bolus. If monensin, during the early postpartum period, increased the
levels of rumen propionate, and therefore increased the levels of blood glucose as
reported by other studies (Richardson et al., 1976; Schelling, 1984), it is reasonable to


63
- Clostridium species: They are not predominant bacteria, but 3 cellulolytic species have
been described. They ferment starch and cellulose. Major products are formate,
butyrate, acetate, C02 and H2.
- Eubacterium species: Gram-positive short rods. They produce acetate from C02 and
H2. They account for 5% of the total bacteria in rumen.
- Lactobacillus species: More common in young animals fed with milk and in older
animals fed concentrates. Major product is lactate.
- Bifidobacterium species: They catabolize glucose, resulting in the production of acetic
and lactic acids.
- Non-sugar-fermenting bacteria: They are a large population of rumen bacterial species
that utilize organic compounds other than sugar. Major products are succinate,
propionate and acetate.
- Large bacteria: Little is known about this family. They ferment small sugars and
produce acids and gases.
- Methanogens: These organisms are not bacteria; in several respects they more closely
resemble eukaryotes. Methane is formed from C02 and H2, but other potential
substrates include formate, acetate, and methanol.
According to Williams and Colleman (1997), in addition to the bacteria in the
rumen there are many larger organisms (5-250 pm) belonging to the group of protozoa.
The largest most obvious and most important protozoa are the ciliates, of which there are
two groups, holotrichs and entodiniomorphs. Despite the metabolic diversity of the ciliates
and their large biomass in the rumen, the protozoa, unlike the bacteria, are not essential for
the survival and development of the host ruminant. However, rumen fermentation is more
stable when an active ciliate population is present. Ciliates are selectively retained in the


45
The effect of ketosis on fertility has been described by Andersson et al., (1991).
They found, in Swedish cattle, that the interval from calving to first service was about 5
days longer in cows with acetone concentrations > 2.0 mmol/L, while the interval to the
last service was shortest at 0.40 to 1.0 mmol/L. The risk for cystic ovaries was markedly
increased (odd ratio = 8.7) in first lactation cows with acetone concentrations > 2.0
mmol/L. In herds with a high incidence of ketosis, primiparous cows had a period 6 days
longer from calving to first service and the period from calving to the last service was 12
days longer than normal cows. Recently, Gillund et al. (2001), found in a Norwegian herd
producing 6,175 kg milk/cow/lactation that 20% of the animals were ketotic as measured
by an Acetone test. The mean days in milk for the first occurrence of ketosis was 29.4.
Cows that did not experience an event of ketosis before first insemination were 1.6 times
more likely to conceive than cows that were ketotic during this early postpartum period.
Ketotic cows had higher BCS than healthy cows before the disease was diagnosed and lost
more BCS than did the latter after ketosis had occurred. Finally, the interval from calving
to conception or number of services per conception did not differ between ketotic and
nonketotic cows. In a Swiss study (Reist et al., 2003), high milk acetone concentration
(>0.4 mmol/L) was associated with 3.2 times higher risk of endometritis. Low plasma
glucose, high serum BHB (> 2.3 mmol/L) and high milk acetone concentrations (> 0.4
mmol/L) during the first week after parturition were indicators of increased risk for ketosis
later during lactation. High BCS (> 3.5; scale 1-5) prior to parturition was associated with
high concentrations of ketones in both milk (acetone > 0.4 mmol/L) and serum (BHB >
2.3 mmol/L). Cows with high BCS prepartum were 3.77 (Cl 95%: 1.63-8.71) times more
likely to have milk acetone levels > 0.4 mmol/L and 6.54 (Cl 95%: 2.0-21.4) times more
likely to have serum BHB levels > 2.3 mmol/L during the first week postpartum.


8
essential for triglyceride assembly (Herdt, 2000). When glucose concentration decreases,
as occurs just after calving, NEFA mobilization from adipose tissue is stimulated (Herdt,
2000; Melendez et al., 2002). When blood glucose levels are sufficient, glucose flow is
favored into the Krebs cycle, therefore its precursors are slowly accumulated in
mitochondria. In liver, the excess of citrate is mobilized out of the mitochondria and
converted to malonyl CoA. This intermediate inhibits the enzyme carnitine acyltransferase
I (CAT I), which inhibits the oxidation of fatty acids and indirectly stimulates TG
synthesis. Contrary, when glucose is low, malonyl CoA decreases, CAT I is activated,
favoring the transport of NEFA into the mitochondria, with the consequent increase in
ketogenesis (Nelson and Cox, 2000).
Endocrine regulation of gluconeogenesis, ketogenesis and lipid metabolism
includes insulin, glucagon, somatotropin, catecholamines, cortisol, thyroid hormones and
leptin (Herdt, 2000, Drackley et al., 2001). Glucose levels in a prepartum dairy cow are
high until parturition (Grum et al., 1996). As a result, insulin concentrations are higher
before calving than after calving and glucagon experiences the opposite pattern (Herdt,
2000; Nelson and Cox, 2000; Drackley et al., 2001). Somatotropin is lower before calving
than post calving with a peak at parturition (Grum et al., 1996). Table 2-2 summarizes the
effect of different hormones on carbohydrate and lipid intermediary metabolism in dairy
cattle.
Calving-Related Disorders
The majority of diseases that affect dairy cows occurs during the peripartum
period, consequently they are also called calving-related disorders (Risco and Melendez,
2002). In general, health disorders present low heritabilities (h2= 0 0.05) and
environmental management plays the most important role in decreasing or preventing their


173
Del Vecchio, R. P., D. J. Matsas, S. Fortin, D. P. Sponenberg, and G. S. Lewis. 1994.
Spontaneous uterine infections are associated with elevated prostaglandin F2 alpha metabolite
concentrations in postpartum dairy cows. Theriogenology 41:413-421.
Del Vecchio, R. P., D. J. Matzas, T. J. Inzana, D. P. Sponenberg, and G. S. Lewis. 1992. Effect
of intrauterine bacterial infusions and subsequent endometritis on prostaglandin F2 alpha
metabolite concentrations in postpartum beef cows. J. Anim. Sci. 70:3158-3162.
Detilleux, J. C., Y. T. Grhn, S. W. Eicker, and R. L. Quaas. 1997. Effect of left displaced
abomasum on test day milk yields of Holstein cows. J. Dairy Sci. 80:121-126.
Dinsmore, R. P., R. Stevens, M. Cattell, M. Salman, and F. Sundlof. 1996. Oxytetracycline
residues in milk after intrauterine treatment of cows with retained fetal membranes. J. Amer.
Vet. Med. Assoc. 209:1753-1755.
Dohoo, I. R., and S. W. Martin. 1984. Disease, production and culling in Holstein-Friesian
cows. IV. Effects of disease on production. Prev. Vet. Med. 2:755-770.
Dohoo, I.R., Martin, W., Stryhn, H. 2003. Screening and diagnostic tests. Pages 101-109 in
Veterinary epidemiologic research. I. Dohoo, W. Martin, H. Stryhn. AVC Inc. Charlottetown,
Prince Edward Island, Canada.
Domecq, J. J., A. L. Skidmore, J. W. Lloyd, and J. B. Kaneene. 1997a. Relationships between
body condition scores and milk yield in a large dairy herd of high yielding Holstein cows. J.
Dairy Sci. 80:101-112.
Domecq, J. J., A. L. Skidmore, J. W. Lloyd, and J. B. Kaneene. 1997b. Relationships between
body condition scores and conception at first artificial insemination in a large dairy herd of
high yielding Holstein cows. J. Dairy Sci. 80:113-120.
Donkin, S. S., M. J. Cecava, and T. R. Johnson. 1998. Protein requirements of transition dairy
cows. J. Dairy Sci. (Midwest Section Abstracts):79.
Donovan, G. A., G. M. DeChant, C. A. Risco, and T. Q. Tran. 1998. Nutritional effects on
laminitis and lameness in dairy cattle. Pages 78 90 in Proc. 35,h Annual Florida Dairy
Production Conference. University of Florida, Gainesville, FL. May 5.
Drackley, J. K. 1999. Biology of dairy cows during the transition period: the final frontier?. J.
Dairy Sci. 82:2259-2273.
Drackley, J. K., D. W. LaCount, J. P. Elliot, T. H. Klusmeyer, T. R. Overton, J. H. Clark, and
S. A. Blum. 1998. Supplemental fat and nicotinic acid for Holstein cows during an entire
lactation. J. Dairy Sci. 81:201-214.


30
uterine horns. This response might be explained for the effect of PGF2alpha on smooth
musculature contraction of the uterus (Melendez et al., 2003c).
Abomasal Disorders
Displacements, dilatations, and volvulus of the abomasum are the most commonly
encountered disorders of the gastrointestinal tract in modem dairy operations (Trent,
1990). Displacement can be on the left side (LDA) or the right side (RDA). (Fecteau et al.,
1999). Omental attachments of the abomasum prevent true torsion around the long axis of
the abomasum, with rotation occurring around an axis through the supporting lesser
omentum. Therefore, a more accurate term for the syndrome is abomasal volvulus,
rather than torsion. Any right-sided displacement that requires further manipulation to free
the pylorus and duodenum may be considered for practical purposes to be a volvulus
(Trent, 1990).
The prevalence of abomasal displacement among dairy herds is variable depending
on geographic location, management practices, climate and other factors (Fecteau et al.,
1999). Left displacement is the most common of the three syndromes, constituting 85 to
95.8% of the total cases of displacements or volvulus (Trent, 1990).
Left displacement of abomasum
In LDA, the abomasum slides under the rumen and dorsally along the left body
wall. The result is a partial impairment of abomasal outflow, leading to abomasal gas
accumulation, electrolyte pooling with subsequent systemic alterations, and depressed
gastrointestinal motility and appetite (Fecteau et al., 1999). A simple case definition is a
cow with decreased appetite accompanied by a progressive decrease in milk production.
An audible, high pitched tympanic resonance (ping) produced by percussion of the left
abdominal wall between the 9th and 12th ribs is a characteristically used to diagnose this


114
Figure 3-5. Least squares means and SEM for test day milk yield by parity 3 or
greater. P < 0.05


CHAPTER 5
EFFECT OF A MONENSIN-CONTROLLED RELEASE CAPSULE ON ENERGY STATUS
IN FLORIDA TRANSITION HOLSTEIN COWS
Introduction
The transition period, defined as the three weeks before and after parturition (Grummer,
1995), is characterized by tremendous physiological and metabolic changes in dairy cows. Cows
experience a certain degree of hypocalcemia with levels of total plasma calcium concentrations <
7.5 mg/dl (Goff, 1999). At calving, non-esterified fatty acids (NEFA) may reach concentrations
> 1.0 mEq/L (Grummer, 1993; Herdt and Gerloff, 1999). Within the first two weeks postpartum
B-hydroxybutyrate (BHB) may reach plasma concentrations > 1000 pmol/L and plasma glucose
< 55 mg/dL (Duffield et al., 1998a; Vallimont et al., 2001). Consequently, if prevention is not
considered during this period cows are at higher risk of developing calving-related disorders (CRD)
(Goff and Horst, 1997b; Kelton et ah, 1998).
Citrus pulp is a common energy concentrate by-product used in dairy cattle diets in
Florida and other states (Arthington et ah, 2002). It contains a high proportion of pectin, which
is not digested by mammalian enzymes, but can be rapidly fermented by ruminal
microorganisms (Hall, 1997).
Monensin is an ionophore that affects rumen fermentation (Van Maanen et ah, 1978),
increasing propionic acid production with a concurrent decrease in the molar proportion of
acetate and butyrate (Richardson et ah, 1976). As a result, monensin has been used to prevent
ketosis and other disorders in dairy cattle (Duffield et ah, 1998a; 1999b; Green et ah, 1999).
140


130
Table 4-4. Least squares means S.E.M. for rumen pH, acetate, propionate, butyrate, valerate,
total VFA, ammonia, D and L-lactate and Acetate:Propionate by time
in multiparous treated and control cows at 10 d pp
._MM 1 4 J t ^r
PH
Ac1
Pr2
Bu3
VFA5
NHj
L-Lac6
D-Lac7
A/P8
mMol/L
M9
6.84
53.2
20.6
9.02
0.92
86.8
1.95
0.03
0.02
2.66
SEM
0.08
3.60
2.17
0.69
0.10
6.60
0.34
0.08
0.12
0.09
TO
c10
7.01
53.4
22.8
8.50
0.84
88.9
2.10
0.04
0.02
2.41
SEM
ii
0.12
5.09
3.07
0.98
0.14
9.31
0.49
0.11
0.17
0.14
6.55
62.3
28.3
11.1
1.47
106.6
4.79
0.20
0.28
2.21
M
0.08
3.60
2.17
0.69
0.10
6.60
0.34
0.08
0.12
0.09
T2
SEM
6.61
62.2
29.3
10.5
1.25
107.2
4.22
0.02
0.03
2.14
C
0.12
5.09
3.07
0.98
0.14
9.31
0.49
0.11
0.17
0.14
SEM
6.48
65.2
27.5
11.5
1.39
108.8
4.28*
0.22
0.30
2.41
M
0.08
3.60
2.17
0.69
0.10
6.60
0.34
0.08
0.12
0.09
T4
SEM
6.52
68.5
32.9
11.9
1.47
119.0
5.01*
0.03
0.05
2.10
C
0.12
5.09
3.07
0.98
0.14
9.31
0.49
0.11
0.17
0.14
SEM
6.46
66.0
27.2
11.6
1.30
109.3
2.80t
0.05
0.04
2.48
M
0.08
3.60
2.17
0.69
0.10
6.60
0.34
0.08
0.12
0.09
T6
SEM
6.48
63.6
29.6
10.9
1.25
109.3
4.0p
0.05
0.02
2.19
C
0.12
5.09
3.07
0.98
0.14
9.31
0.49
0.11
0.17
0.14
SEM
* (P < 0.15)
f (P < 0.05)
1 Acetate,2 Propionate,3 Butyrate,4 Valerate,5 Total Volatile Fatty Acids,6 L-Lactate,7 D-
Lactate,8 Acetate to Propionate Ratio, 9 Monensin, 10 Control, 11 Standard Error of Mean, T0=
time 0, 7:00 A.M. before feeding; T2= time 2, 2 h after A.M. feeding; T4= time 4, 4 h after
A.M. feeding; T6= time 6, 6 h after A.M. feeding.


51
To prevent ketosis-fatty liver complex, fat cows at parturition should be avoided.
Dry cows should be fed a diet to maintain weight, not to lose weight (Gerloff and Herdt,
1999). Cows overfed during the dry period have higher concentrations of plasma
postpartum NEFA as a result of greater lipolysis after parturition. Cows experiencing
more severe negative energy balance results in a high liver TG concentration (Grummer,
1993; Rukkwamsuk et al., 1998, Rukkwamsuk et al., 1999a). Rukkwamsuk et al. (1999a)
demonstrated that cows overfed before parturition, had lower percentages of oleic acid and
higher percentages of linoleic acid than normal cows in liver, but percentages of other
fatty acids were similar. One week after parturition the percentages of palmitic and oleic
acids were higher than one week before parturition. They concluded that the increased
lipolysis after calving increased the hepatic TG concentration and shifted hepatic fatty acid
composition. Kaneene et al. (1997) showed that metabolic events related to negative
energy balance were related to increased risk of metritis and RFM. Higher energy
consumption during the last weeks of the dry period (more grain content of the ration) was
related to reduced disease risk at parturition. Finally, they concluded that serum NEFA
and cholesterol concentrations have potential as indicators of disease risk in dairy cows.
Scoring the body condition of the cows at dry-off and managing the nutritional
program of transition cows is a useful tool to avoid fatty liver development and related
problems (Gerloff and Herdt, 1999). Two adequate body condition score charts are
available for Holstein dairy cattle. One chart was developed by Edmonson et al. (1989)
and the other was developed by Ferguson et al. (1994). It has been demonstrated that high
body condition score at parturition negatively affects reproductive parameters, milk yield
and health of the cows during the current lactation (Butler and Smith, 1989; Domecq et
al., 1997a; Domecq et al., 1997b; Markusfeld et al., 1997; Heuer et al., 1999).


54
feed intake in pregnant Holstein heifers 90 days prior to calving. The results showed that
propylene glycol linearly increased glucose and insulin and decreased BHB and NEFA in
blood. They concluded that the dose of 296 ml of propylene glycol was almost as effective
as a dose of 887 ml in reducing lipid mobilization during restricted feed intake. After that,
the same research group (Christensen et al., 1997) demonstrated that administration of
propylene glycol as an oral drench or mixed with concentrate resulted in higher serum
insulin and lower plasma NEFA concentrations than did feeding propylene glycol as part
of a TMR system. Finally, Laranja da Fonseca et al. (1998) found that cows fed 300 mg/d
propylene glycol orally had an increased milk yield of 2.5 kg/day, glucose was similar,
BHB was lower by almost 2 mg/dl and NEFA were lower by 0.031 mEq/L than cows
without the supplement; however these differences were not statistically significant.
Propionate is produced in the rumen after fermentation of starch, fiber, and protein.
It is the major glucose precursor in ruminants that are in positive energy balance and is the
major substrate used for gluconeogenesis, followed by lactate and amino acids. Its
contribution to total glucose production has been consistently measured at 30 to 50%.
Most of the propionate absorbed from the rumen is metabolized in the rumen epithelium
or liver. (Herdt, 1988). A small portion escapes hepatic and rumen epithelial metabolism,
passes into the systemic blood circulation, and is extracted by peripheral tissues (Miettinen
and Huhtanen, 1996).
After being absorbed by the rumen epithelial wall, propionate is transported via
the portal vein to the liver where it is converted to glucose via pyruvate and oxaloacetate.
Propionate decreases the ruminal absorption of acetate and butyrate. Propionate is
incorporated into the Krebs cycle through succinyl coenzyme A and is converted to


ELANCO Animal Health, USA and Mr. Leo Richardson and Richard Tucker.
Dr. Todd Duffield for his comments and for sending me a copy of his
dissertation.
Dr. Julian Bartolom, for his unconditional friendship.
Dr. Victor Monterroso, for his valuable comments, loyalty, and support during
the most difficult part of my staying in USA.
Rural Animal Medicine Service, Dr. Owen Rae, chairman of RAMS, Delores,
residents, internships and OPS students for their help.
Shelly Lanhart, Chris Sissle and Christina Herejk for their help on my program
and the lab analysis.
Friends from different countries for their friendship (Gina, Antonio, Martin,
Billy, etc).
My friends in Chile for their continuous contact with me during this time and
their friendship.
My brother, grandmother, nieces and nephew for their support and feelings
about us.
My father, Oscar and my mother, Eliana, for their entire life of efforts, and
affection.
My wife, Maria Ester, for her support and love and for being a wonderful and
great woman, mother and wife.
Ignacio, Diego and Elisa, for their love and for being the most important reason
to be here.
IV


93
Cows were dried-off between 50 to 70 days before expected parturition (BEP) and
housed in a dry-lot (Far-off cows). They were fed a typical Florida dry cow diet (Table
3.1 and 3.2). At 21 days BEP, dry cows were moved from the far-off pen to a different
dry-lot with adequate feed-bunk space and shade (close-up cows). Twice a day, they
received a diet containing citrus pulp with a DCAD of -51.7 mEq/kg DM using the
equation DCAD (mEq)= (Na + K) ( Cl + S) (Tables 3.1 and 3.2).
Cows calved in the close-up pen and the calf were immediately separated from
the dam at parturition. If the cow needed calving assistance she was moved to a
maternity bam. Cows were processed within 12 h postpartum on a routine basis which
included recording BCS, udder score (for presence of edema), reproductive tract status
(trauma or lacerations), and whether the cow was suspected of having RFM. If cows
developed either RFM or milk fever they were treated and remained in the hospital bam
until recovery. After postpartum processing, cows were moved to a postpartum lot and
fed a diet higher in forage NDF (Tables 3.1 and 3.2).
Beginning 60 days postpartum, cows received bST (Posilac, 500 mg
sometribove zinc, subcutaneously; Monsanto, St Louis, Missouri) every 14 days during
the entire lactation.
Reproductive management consisted of a voluntary waiting period of 80 days
until first insemination. First service was synchronized and subjected to a timed artificial
insemination protocol. Thereafter, cows were identified in estrus using either visual
observation or a computerized pedometer system (Afimilk, S.A.E. Afikim, Kibbutz
Afikim, 15148, Israel) and bred on heat detection. Pregnancy diagnosis was performed
by herd veterinarians by palpation per rectum of the uterus and its contents at
approximately 42 to 49 d after insemination.


42
reproductive performance was also accounted for in the model. Milk yield had a
significant effect on culling decisions, depending on the stage of lactation. In late lactation
the highest producers were at lowest risk of being culled and the lowest producers had the
highest risk. Milk yield and parity were interrelated in their effects on culling (Rajala-
Schultz and Grdhn, 1999a; 1999b).
Ketosis and fatty liver
Ketosis is defined as a metabolic disease characterized by high levels of ketone
bodies affecting cattle, sheep and goats. Ketosis affects dairy cows in the period from
parturition to 6 weeks postpartum (Herdt and Gerloff, 1999). There are two types of
ketosis, primary and secondary. Cattle with primary clinical ketosis havea decreased
appetite and elevated serum, milk, urine or breath ketones in the absence of another
concurrent disease (Kelton et al., 1998).
The ketone bodies, acetone, acetoacetate, and B-hydroxybutyrate (BHB), are
formed in the liver during oxidation of fatty acids (Nelson and Cox, 2000). Acetone, a 3
carbon compound, is produced in small quantities and is exhaled to the environment. The
first produced ketone body is acetoacetate (4 carbons), which either is reversibly reduced
to BHB in mitochondria, or is enzymatically or spontaneously decarboxylated to acetone
(Nelson and Cox, 2000).
Ketosis may be clinical or subclinical. Subclinical ketosis is defined as a
preclinical stage characterized by elevated blood ketone body concentrations without
clinical signs such as loss of appetite, hard feces, or dullness (Anderson, 1988; Duffield,
2000; Geishauser et ah, 2001).
The lactational incidence rate of clinical ketosis, based on 36 citations from 1979
to 1995, ranged from 1.3% to 18.3%. The median lactational incidence rate was 4.8%


89
Table 2-10. Minimum trace mineral and vitamins requirements for dry, and prepartum
cows
Nutrient
Far-off dry cows
Close-up dry cows
Cobalt (ppm)
0.11
0.11
Copper (ppm)
16
16
Iodine (ppm)
0.4
0.4
Iron (ppm)
26
26
Manganese (ppm)
22
22
Selenium (ppm)
0.30
0.30
Zinc (ppm)
30
30
Vitamin A (IU/kg)
5500
6500
Vitamin D (IU/kg)
1500
1700
Vitamin E (IU/kg)
80
88
Adapted from: National Research Council. 2001. Nutrient requirements of dairy cattle, 7th
revised edition. Washington, D.C.: National Academy Press.


174
Drackley, J. K., T. R. Overton, and G. N. Douglas. 2001. Adaptations of glucose and long-
chain fatty acid metabolism in liver of dairy cows during the periparturient period J Dairv
Sci. 84(E. Suppl.):E100-El 12.
Duffield, T. 1997. Effects of a monensin controlled released capsule on energy metabolism,
health, and production in lactating dairy cattle. Thesis dissertation, University of Guelph,
Guelph, ON, Canada.
Duffield, T. 2000. Subclinical ketosis in lactating dairy cattle. Vet. Clin. North Am. Food
Anim. Pract. 16:231-253.
Duffield, T. F., R. Bagg, L. DesCoteaux, E. Bouchard, M. Brodeur, D. DuTremblay, G.
Keefe, S. LeBlanc, and P. Dick. 2002. Prepartum monensin for the reduction of energy
associated disease in postpartum dairy cows. J. Dairy Sci. 85:397-405.
Duffield, T. F., S. LeBlanc, R. Bagg, K. E. Leslie, J. Ten Hag and P. Dick. 2003. Effect of a
monensin controlled release capsule on metabolic parameters in transition dairy cows. J.
Dairy Sci. 86:1171-1176.
Duffield, T. F., K. E. Leslie, D. Sandals, K. Lissemore, B. W. McBride, J. H. Lumsden, P.
Dick, and R. Bagg. 1999a. Effect of prepartum administration of monensin in a controlled-
release capsule on milk production and milk components in early lactation. J. Dairy Sci.
82:272-279.
Duffield, T. F., K. E. Leslie, D. Sandals, K. Lissemore, B. W. McBride, J. H. Lumsden, P.
Dick, and R. Bagg. 1999b. Effect of a monensin-controlled release capsule on cow health and
reproductive performance. J. Dairy Sci. 82:2377-2384.
Duffield, T. F., D. Sandals, K. E. Leslie, K. Lissemore, B. W. McBride, J. H. Lumsden, P.
Dick, and R. Bagg. 1998a. Effect of prepartum administration of monensin in a controlled-
release capsule on postpartum energy indicators in lactating dairy cows. J. Dairy Sci. 81:2354-
2361.
Duffield, T. F., D. Sandals, K. E. Leslie, K. Lissemore, B. W. McBride, J. H. Lumsden, P.
Dick, and R. Bagg. 1998b. Efficacy of monensin for the prevention of subclinical ketosis in
lactating dairy cows. J. Dairy Sci. 81:2866-2873.
Duvfa, G. S., E. E. Bartley, A. D. Dayton, and D. O. Riddell. 1983. Effect of niacin
supplementation on milk production and ketosis of dairy cattle. J. Dairy Sci. 66:2329-2336.
Dyk, P. B., R. S. Emery, J. L. Liesman, H. F. Bucholtz, and M. J. VandeHaar. 1995.
Prepartum non-esterified fatty acids in plasma are higher in cows developing periparturient
health problems. J. Dairy Sci. 78(Suppl.l):264.
Edmonson, A. J., I. J. Lean, L. D. Weaver, T. Farver, and G. Webster. 1989. A body condition
scoring chart for Holstein dairy cows. J. Dairy Sci. 72:68-78.


158
Blood and Milk Sampling
At 14 days postpartum from each experimental cow, a composite milk sample was
obtained from the milking unit at the parlor during the morning milking (7:00 a.m. to 3:00
p.m.). Milk was collected in 50 ml plastic tubes, placed in a container with ice packs and
carried to the farm laboratory facilities. Immediately after milking, the cow was separated
and placed in a chute. In a random sub-sample of 50 cows per group a blood sample for
BHB determination was taken from the coccygeal vein, using an evacuated tube without
anticoagulant (Vacutainer, Becton Dickinson, Rutherford, NJ).
Laboratory Analysis
Milk samples were tested for BHB using a milk ketone body test strip (Ketolac00,
Nagoya, Japan). The strip is introduced in the milk sample and is interpreted 3 minutes
later. The result is a semi quantitative colorimetric enzymatic reaction based on the
conversion of BHB to acetoacetate by the enzyme fi-hydroxy butyrate dehydrogenase
(Nelson and Cow, 2000). A color scale defines 0, 50, 100, 200, 500 and 1000 pmol/L of
BHBA. Two-hundred pmol/L of BHB is considered a positive reaction (subclinical ketosis).
Blood samples were centrifuged at 4000 rpm for 10 minutes. Serum was separated and
stored in plastic tubes and frozen at 20C until analysis was performed.
Serum BHB was based on an enzymatic-colorimetric method (Williamson and
Mellanby, 1974), using a commercial kit (Pointe Scientific, Inc. BHA Set. Lincoln Park,
MI, USA).
Statistical Analysis
The null hypothesis was that there is no difference in the concentration of serum
BHB metabolites or the proportion of cows with a positive milk ketone test between


52
Niacin is a vitamin belonging to the B group. As an additive it can reduce lipid
mobilization during early lactation, minimize incidence of ketosis and improve milk yield
and fat synthesis (Hutjens, 1991). In several studies in which 6 to 12 g/day of niacin was
supplemented to dairy cows fed diets with added fat, niacin increased milk production and
concentration of milk fat and protein. Niacin or nicotinic acid has been also suggested as a
treatment or preventive for fatty liver in dairy cows during the prepartum period; but there
is little research evidence to confirm its use as a preventive agent for fatty liver (Gerloff
and Herdt, 1999). Niacin is antilipolytic and may decrease plasma concentrations of
NEFA and BHB (DufVa et al., 1983). Nevertheless, in another study, niacin either alone or
in combination with high dietary non-fiber carbohydrates (NFC), did not influence
lactation performance or metabolic status of cows. Increased dietary NFC significantly
lowered plasma NEFA and BHB concentrations, and significantly elevated liver glycogen
(Minor et al., 1998). In another study, Drackley et al. (1998) determined that nicotinic acid
increased NEFA when it was added to a control diet but decreased NEFA when it was
added to a diet supplemented with fat. Although data seem to be controversial with respect
to niacin in transition cows, the addition of 6 to 12 g/d of niacin to diets during the first 2
to 3 month of lactation has been recommended. Heavier dairy cows (BCS > 3.5) have
shown optimal response; thin cows are less responsive. Niacin should be supplemented 2-
3 weeks before parturiton to 10-12 weeks after parturition, but it should be analyzed on a
cost vs response basis (Hutjens, 1992).
Propylene glycol is a glucogenic compound used to treat ketosis in postpartum
dairy cows (Studer et al., 1993). A high proportion of propylene glycol by-passes the
rumen and is absorbed in the small intestine; the rest is metabolized to propionate.
Ruminal molar percentage of acetate decreased and acetate to propionate ratio also


87
80%. Clean water should be provided expecting cows to drink 2 liters for each 0.45 kg of
milk. Enough feed bunk space should be also provided.
Cows should peak in milk production 8 to 10 weeks after calving. First-calf heifers
should peak within 75% the production of older cows. For each extra 1 kg of milk at peak
production the average cow will produce 200 to 220 kg more milk for the entire lactation
period. Milk proteimfat ratio should be near 0.85-0.88 for Holstein cows. Forage intake
using a good quality roughage should be maximized.
Energy density of the top cows should be 1.76 Mcal/kg of EN1. Non fiber
carbohydrates levels should be between 35 and 42% with 25 and 35% starch in the total
ration.


61
According to Stewart et al. (1997) the major groups of rumen bacteria are:
Prevotella species (formerly Bacteroides ruminicola): strictly anaerobic, gram
negative bacteria. They degrade starch, xylans and pectins. Fermentation products
include acetate, succinate and propionate. They are sensitive to monensin, but
mutations to resistance occur readily.
Ruminobacter (Bacteroides) amylophilus: Rare, but it appears to be the predominant
starch digester. It also possesses significant proteolytic activity. Major fermentation
products are acetate and succinate.
Fibrobacter (Bacteroides) succinogenes: One of the most widespread celluloytic
bacteria of the rumen. Major fermentation products are acetate and succinate.
Succinovibrio dextrinosolvens: Gram-negative rod ferments pectins and dextrins.
Major products are acetate and succinate.
Succinimonas amylolytica: Gram-negative bacteria associated with starch digestion.
Major products are acetate and propionate.
Spirochaetes: The account for 1% to 6% of the total viable bacteria count in bovine
rumen liquor. The rumen spirichaetes are predominantly treponemes. They ferment
pectins and small sugars. Major products are acetate, formate and succinate.
Anaerovibrio lipolytica: Gram-negative rods that hydrolyze lipids and ferment
fructose, glycerol and lactate. Major products are propionate, succinate, lactate, CO2
and H2.
Selenomonas ruminantium: This gram-negative curve rod constitutes 22-51% of the
total viable count. Some strains ferment starch, others ferment small sugars, glycerol
or lactate. Another important activity of S. ruminantium is the decarboxylation of
succinate to propionate. Major products are lactate, propionate and acetate. They
appear to be relatively resistant to monensin.


CHAPTER 3
EFFECT OF A MONENSIN-CONTROLLED RELEASE CAPSULE ON THE
INCIDENCE OF CALVING RELATED DISORDERS, FERTILITY AND MILK
YIELD IN FLORIDA HOLSTEIN TRANSITION COWS
Introduction
The transition period in dairy cows is defined as the three weeks before and three
weeks after parturition and is characterized by tremendous physiological and metabolic
demands where calving is a major component of this stage (Grummer, 1995).
Consequently, calving-related disorders (CRD) such as milk fever (MF), retained fetal
membranes (RFM), metritis, fatty liver, ketosis, left and right displacement of
abomasum (DA), mastitis, and lameness are common conditions, that have a negative
economic impact on dairy herds (Goff and Horst, 1997b; Kelton et al., 1998). The
negative impact of CRD on reproductive performance and milk production has been
well documented (Rajala-Schultz and Grohn,1998; Rajala-Schultz et al., 1999;
Fourichon et al., 1999; Grdhn et al., 1999). Therefore any attempt to decrease the
incidence of CRD will positively affect milk yield and fertility in dairy cows.
Citrus pulp is an energy concentrate by-product produced in subtropical regions,
of which south central Florida remains the largest area of production and it is a common
component of dairy cattle diets in Florida and other southern states (Arthington et al.,
2002). Citrus pulp is predominantly composed of pectin, which is indigestable by
mammalian enzymes, but can be rapidly fermented by ruminal microbes (Hall, 1997).
Monensin is an ionophore that affects rumen fermentation patterns (Van Maanen
et al., 1978). The most consistent and well-documented fermentation alteration observed
91


103
suggest that this mechanism is plausible to explain changes in milk production. The long
term mechanism might be explained by the positive effect of monensin on BCS at
calving. Therefore, cows might experience an enhanced energy status during the entire
lactation. Indeed, cows in fair BCS at calving are higher producers than thinner and
obese cows (Duffield et al., 1999a). In addition, phenotypic correlations between BCS at
calving and milk yield are slightly positive in Holstein cows (Dechow et al., 2001).
Monensin consistently decreased milk fat % during the entire lactation. These
results are partially in accordance with previous studies (Hayes et al., 1996; Van Der
Werf et al., 1998; Vallimont et al., 2001). Since this effect was observed during the
entire lactation and only on third parity or older cows, it is reasonable to suggest that this
difference might be due to a dilution effect of higher milk production in the same group
of animals.
Reproductive Responses
Monensin had a tendency to improve conception rate at first service (P=0.12).
The rest of the variables were not significantly different. There are no previous studies
reporting positive effects of monensin on reproductive responses (Lean et al., 1994;
Hayes et al., 1996; Duffield et al., 1999b). An explanation for this controversy is the
multifactorial nature of fertility and its low heritability values in dairy cows (Weigel and
Rekaya, 2000; Veerkamp et al., 2001). In the present study, unlike other studies
(Duffield et al., 1998a, 1998b, 1999a; 1999b), treated and control cows were handled
very homogeneously. They were housed in the same environment and were fed the same
diet during the same period of time. Reproductive management was exactly the same
and one large source of variation was the application of the bolus. Therefore, under these
conditions the tendency for better CR at first service in cows treated with monensin


31
condition (Kelton et al., 1998). Feces are often softer than normal. Rectal temperature,
respiratory and heart rate are generally normal (Fecteau et al., 1999).
Left displacement of the abomasum occurs most commonly two weeks pre- to 8
weeks postpartum. The incidence of LDA, based on 22 citations from 1982 to 1995
ranged from a postpartum incidence rate of 0.3% to 6.3%. The median incidence rate was
1.7% (Kelton et al., 1998). Detilleux et al., (1997), in New York State, determined an
incidence of LDA that ranged from 2.1 to 8.7%. The mean number of days of lactation at
which LDA was diagnosed was 20.6. In New York state, Grdhn et al., (1998) observed a
lactational incidence risk of LDA of 5.3%. Cameron et al., (1998) in 1170 multiparous
Holstein cows from 67 high producing dairy herds in Michigan, found an incidence of
LDA of 6% in primiparous and 7% in multiparous cows. Cows with twins presented an
incidence of 11 to 12 %. 0stergaard and Grohn (1999) in three Danish research herds
(4414 lactations) determined an incidence of LDA in first lactation cows of 0.6% and in
older cows 1.2%, with a first diagnosis at 19 days in milk (mean) or 17 days in milk
(median). In Florida, Massey et al, (1993) and Melendez et al. (2003b) found an LDA
incidence of 2.4% and 3.9%, respectively.
Economic losses from LDA include lost milk production and the cost of the
surgery which is estimated to total approximately $340 per case (Guard, 1994; Kelton et
al., 1998). The effect of LDA on test day milk yields of Holstein cows have been studied
by several authors. Detilleux et al., (1997) in New York State and using 12,572 Holstein
cows between parity 1 and 6, determined that from calving to 60 days after diagnosis,
cows with LDA yielded on average 557 kg less milk than did cows without LDA. Thirty
percent of losses occurred before diagnosis and milk loss increased as parity and
productivity increased. In each parity, the lactation curves of cows with LDA were


143
Blood Sampling
Blood samples were taken from the coccygeal vein into an evacuated tube without
anticoagulant for serum, heparinized tubes for plasma metabolites and NaF for glucose analysis
(Vacutainer, Becton Dickinson, Rutherford, NJ). Samples were collected on the day of
assignment (50 to 70 days BEP) before receiving any treatment, on day 21 BEP, at calving, and
at 7, 14, and 21 d after calving at the same time of the day.
Laboratory Analysis
Blood samples were centrifuged at 4000 rpm for 10 minutes. Plasma and serum were
separated and stored in plastic tubes and frozen at 20C until analysis was performed.
Serum NEFA, plasma BHB and serum glucose were measured. Nonesterified fatty acids
were determined by an enzymatic-colorimetric method, using a commercial kit (NEFA-C kit
WAKO, Osaka, Japan). Beta-hydroxybutyrate was determined by an enzymatic-colorimetric
method, using a commercial kit (Pointe Scientific, Inc. BHA Set. Lincoln Park, MI, USA).
Glucose was determined using an enzymatic colorimetric method (Mutarotase GOD) (Autokit
Glucose, WAKO, Osaka, Japan). Absorbance were measured in a microplate reader (Dynex,
MRX Revelation, Chantilly, VI, USA) using the corresponding length waves for each
particular metabolite.
Statistical Analysis
The null hypothesis was that there is no difference in the concentration of blood
metabolites between treatment groups. Blood metabolites and test day milk yield were analyzed
constructing mixed models for repeated measures. Body condition score at calving,
accumulated real and ME305 milk yield were analyzed by ANOVA mixed model. Variables


26
Uterine infections are one of the most frequent disorders affecting dairy cows
during the post partum period (Youngquist and Shore, 1997). They are a major cause of
economic losses to the cattle industry being related with systemic disease, decrease
reproductive efficiency, reduced milk production, increased replacement costs and
reduced genetic progress (Kelton et al., 1998). The total cost to producers for each
lactating dairy cow with a uterine infection was estimated at $106 (Barlett et ah, 1986a).
In a review by Fourichon et ah, (1999), only two studies out of ten showed losses in milk
production between 100 to 270 kg per lactation in cows with metritis. Typically, no milk
loss was associated with metritis. Rajala-Schultz and Grhn (1998) using data from
37,776 Finish Ayrshire dairy cows determined that when metritis was treated as one
disease complex, regardless of the time of its occurrence, metritis had no significant effect
on milk yield. However, when early and late metritis were analyzed separately, the time of
disease occurrence had an effect on milk yield. Late metritis was not associated with milk
loss yet early metritis accounted for 46.2 kg less milk less in cows with metritis within 28
days after calving.
Barlett et ah (1986a) in Michigan herds determined that cows with metritis were
1.3 times more likely to be culled than were cows without metritis. A more recent study in
Holstein cows in New York State demonstrated that metritis had no effect on the risk of
culling. In contrast, other diseases such as mastitis, MF, RFM, LDA, ketosis and ovarian
cysts significantly affected culling at different stages of lactation (Grhn et ah, 1998).
The diagnosis of metritis has been very subjective and has been applied to clinical
conditions that range from cows that are nearly normal to those affected by severe, life
threatening sepsis (Youngquist and Shore, 1997). Metritis has been defined as an
inflammation of all layers of the uterus typically developed within a few days to several


57
Table 2-6. Ketone body field test comparison
Product
Ketone body
Sample
sen(1)
spec(2)
PV(+)(J)
PV(-)(4)
'Ketolac5
BHB
Milk
>50
(pmol /L)
91.9
54.9
28.8
97.2
> 100
72.4
89.4
42.7
94.3
>200
44.8
96.8
73.6
89.8
>500
17.2
100
100
85.9
> 1000
3.4
100
100
84.0
Ketostix(t)
Acetoacetate (*)
Urine
4.6
100
100
84.2
Ketocheck (t)
Acetoacetate (*)
Milk
27.6
100
100
87.5
Utrecht
Acetoacetate (*)
Milk
42.5
100
100
89.8
powder(t)
Bioketone (f>
Acetoacetate (*)
Milk
33.3
99.8
96.7
88.4
Pink Test(#)
Acetoacetate
Milk
>100
(pmol /L)
76
93
60
96
>300
38
98
78
92
Ketolac(#)
BHB
Milk
>50
(pmol /L)
91
56
21
98
>100
80
76
30
97
>200
59
90
43
94
Uriscan(#)
Acetoacetate
>500
(pmol /L)
Milk
13
100
0
90
Rapignost{lt)
Acetoacetate
Milk
3
100
0
91
>500
(pmol /L)
(1) : Sensitivity,
(2) : Specificity
(3) : Positive Predictive Value
(4) : Negative predictive Value
(t): Geishauser et al., (1998).
(#): Geishauser et al., (2000).
(*): Qualitative test (yes/no)


127
Table 4-1. Diet composition of dry cow far-off, dry cow transition and lactating
transition cows
Feed
Dry cows
far-off
Dry-cows
transition
Lactating
transition
Alafalfa hay
-
18.00
10.54
Coastal hay
16.82
2.24
10.08
Com silage
22.17
25.60
28.32
Ryegrass silage
22.50
9.88
-
Com hominy
7.22
9.14
16.96
Citrus pulp
5.20
7.18
7.84
Soybean meal 48
5.56
8.68
12.35
Wet brewers grain
8.79
7.71
5.61
Soy hull pellet
8.30
7.28
5.40
Lactowhey 1
1.93
-
-
Springer minerals
-
4.29
-
Dry cow minerals
l.5l
-
-
Lactating minerals
1 A x J ..
-
-
2.90
1 Ammoniated whey (61.5% CP)


162
studies did not use diets based on citrus pulp, the present findings are in accordance with the
Canadian and Australian research using monensin as a bolus (Abe et al., 1994; Stephenson
et al., 1997; Van Der Werf et al., 1998; Duffield et al., 1998b; Green et al., 1999; Duffield
et al., 2003). Therefore, because pectin-fermenting bacteria are gram-negative monensin-
resistant bacteria (Nagaraja et al., 1997; Stewart et al., 1997), monensin still improved the
energy status of transition cows with a lower rate of ketogenesis than control cows.
Milk yield at 14 d pp and BCS at calving were predictors of serum BHB
concentrations (P < 0.05). Both predictors were related positively with BHB in serum. The
higher the milk yield at 14 d postpartum and the higher the BCS at calving the higher the
concentration of BHB in serum at d 14 postpartum. However, milk yield at 14 d pp and
BCS at calving were not associated with a positive result of milk BHB (Table 6.5) (P >
0.05). This might be explained because the milk ketone test is only a semi-quantitative
colorimetric method.
It is concluded that a monensin-slow release capsule inserted at dry-off in cows fed
citrus pulp decreased the proportion of cows developing subclinical ketosis at 14 d pp, both
using serum and milk BHB determination.


5
Physiological and Metabolic Changes during the Transition Period
Dry Matter Intake
Dry matter intake (DMI) is a function of animal and dietary factors affecting
hunger and satiety (Allen, 2000). Dry matter intake starts to decrease a few weeks before
parturition with the lowest level occurring at calving (Ingvartsen and Andersen, 2000).
Average values for the preffesh transition period have been reported to range between 1.7
and 2.0% of body weight (BW) (Hayirli et al., 1999). However this is not a constant value
and it can be influenced by the ration that is fed (concentration of nutrients), the stage of
the transition period, body condition score (BCS) and parity (Hayirli et al., 2002). Dry
matter intake decreases about 32% during the final three weeks of gestation, and 89% of
that decline occurs at five to seven days before calving (Hayirli et al., 2002). Most cows
rapidly increase DMI for the first three weeks after calving (Ingvartsen and Andersen,
2000). As a percentage of body weight, heifers consume less feed than cows at 21 days
before calving. By the time of calving, intake is more similar (Table 2-1) (Hayirli et al.,
1999).
Table 2-1. Predicted changes in dry matter intake for Holstein dairy cows duringthe last 3
weeks prior to calving
Cows
DMI (Kg/d)
DMI, % BW
% Change
D-21
D-l
D-21
D-l
Heifer (605 kg)
10.3
7.4
1.71
1.22
-28
Cow (740 kg)
14.4
9.8
1.94
1.33
-32
Adapted from: Hayirli, A., R. R. Grummer, E. Nordheim, P. Crump, D. K. Beede, M. J.
VandeHaar, L. H. Kilmer,, J. K. Drackley, D. J. Carroll, G. A. Varga, and S. S. Donkin.
1999. Prediction equations for dry matter intake of transition cows fed diets that vary in
nutrient composition. J. Dairy Sci. 82(Suppl. 1):113.
The National Research Council (2001) reported that DMI as percentage of body
weight during the last 21 days of gestation for heifers is equal to 1.71 0.69 e 0 35t


133
Table 4-7. Least squares means S.E.M. for serum 13-hydroxy butyrate, non-esterified fatty
acids and glucose by time of sampling in primiparous treated and control cows
between dry-off and 21 d pp
Time
Group
BHB
mmol/L
NEFA
meq/L
Glucose
mg/dL
0
M
0.21 0.24
0.25 0.27
72.90 9.15
C
0.23 0.14
0.48 0.15
67.16 5.28
1
M
0.57 0.24
1.050.27t
74.79 9.15
C
0.55 0.14
1.16 0.15t
72.62 5.27
2
M
0.52 0.24n
0.81 0.27
58.59 9.15
C
0.90 0.13t+
1.24 0.15
60.01 4.89
3
M
0.68 0.24
0.78 0.27*
56.85 9.15
C
1.00 0.14
1.420.15t
52.87 5.27
4
M
0.36 0.24*
0.44 0.27t+
53.03 9.15
C
0.90 0.14t
0.91 0.15**
50.21 4.89
5
M
0.51 0.24
0.57 0.27
59.68 9.15
C
0.71 0.14
0.64 0.15
56.57 5.28
n (P <0.1)
f (p < 0.05)
0: assignment, dry-off; 1: 21 d before expected parturition; 2: Calving; 3: 7 d pp; 4: 14 d pp 5
21 d pp


191
Youngquist, R.S.; and M. D. Shore. 1997. Postpartum uterine infecctions. Pages 335-340 In
Current therapy in large animal theriogenology. R.S. Youngquist. W.B. Saunders Company.
Philadelphia, PA.


Abstract of Dissertation Presented to the Graduate School of the University of Florida in
Partial Fulfillment of the Requirements for the Degree of
Doctor of Philosophy
EFFECTS OF MONENSIN IN TRANSITION HOLSTEIN DAIRY COWS FED DIETS
CONTAINING CITRUS PULP
By
Pedro G. Melendez
May 2004
The objective of this research was to determine the effect of monensin-controlled
release capsule on the incidence of periparturient disorders, cow performance, and blood
metabolites in Holstein cows fed diets containing citrus pulp. Four studies were con
ducted on a dairy farm located in Florida. In all four studies, cow were assigned
randomly at dryoff (50 to 70 days before expected parturition [BEP]) to one of two
groups. The treated group received orally a monensin capsule (CRC Rumensin,
ELANCO Animal Health, Guelph, ON) that releases 300 mg of monensin per day for 95
days. Control cows were matched by parity and received no treatment. Studies 1, 2, and 3
began between July and August 2001. In study 1, 580 cows were assigned to two groups.
Outcome variables were incidence of periparturient disorders, milk yield and solids, body
condition score (BCS) at calving, and reproductive responses. In study 2, 60 cows were
randomly assigned to two groups. Outcome variables were milk yield and BCS at
calving. Blood samples were collected at assignment; 21 d BEP; at calving; and at 7, 14,
and 21 d after calving. In study 3, 300 cows were assigned similarly. At 14 days
postpartum, a milk sample was obtained for beta hydroxyl butyrate (BHB). In a
Xll


190
Varga, G. A., H. M. Dann, V. A. Ishler. 1998. The use of fiber concentrations for ration
formulation. J. Dairy Sci. 81:3063-3074.
Vazquez-Aon, M., S. Bertics, M. Luck, R. R. Grummer and J. Pinheiro. 1994. Peripartum
liver triglyceride and plasma metabolites in dairy cows. J. Dairy Sci. 77:1521-1528.
Veerkamp, R.F., E.P.C. Koenen, G. de Jong. 2001. Genetic correlations among body
condition score, yield, and fertility in first-parity cows estimated by random regression
models. J. Dairy Sci. 84: 2327-2335.
Waltner, S.S., J.P. McNamara, and J.K. Hillers. 1993. Relationships of body condition score
to production variables in high producing Holstein dairy cattle. J. Dairy Sci. 76:3410-3419.
Wells, S., A. Trent, W. Marsh, and R. Robinson. 1993. Prevalence and severity of lameness in
lactating dairy cows in a sample of Minnesota and Wisconsin herds. J. Amer. Vet. Med.
Assoc. 202:78-82.
Wedegaertner, T.C., and D.E. Johnson. 1983. Monensin effects on digestibility,
methanogenesis and heat increment of a cracked com-silage diet fed to steers. J. Anim. Sci.
57:168-185.
Weigel, K.A., R.G. Rekaya. 2000. Genetic parameters for reproductive traits of Holstein cattle
in California and Minnesota. J. Dairy Sci. 83: 1072-1080.
Williams, A.G. and G.S. Coleman. 1997. The rumen protozoa. Pages 73-139 in The rumen
microbial ecosystem. Second edition. P.N. Hobson and C.S. Stewart. Blackie Academic &
Professional
Williams, C.B., P.A. Oltenacu, and C.J. Sniff. 1989. Refinements in determining the energy
value of body tissue reserves and tissue gains from growth. J. Dairy Sci. 72:264-269.
Williams, E.L., M.M. Pickett, L.C. Griel, K.S. Heiler, G.A. Varga, and S.S. Donkin. 2003.
Effect of prepartum dietary carbohydrate source and monensin on expression of gluneogenic
enzymes in liver of transition dairy cows. J. Dairy Sci. 86(Suppl. 1):M180. (Abstr.)
Williamson, D. H. and J. Mellanby. 1974. D-3-hydroxybutyrate. Page 1836 in Methods of
enzymatic analysis. H. U. Bergmeyer, ed. Academic Press, London, England.
Yang, C.M.J., and J.B. Russell. 1993. The effect of monensin supplementation on ruminal
ammonia accumulation in vivo and the numbers of amino acids-fermenting bacteria. J. Anim.
Sci. 71:3470-3476.
Youngquist, R. S., W. F. Braun. 1993. Abnormalities of the tubular genital organs. Vet. Clin.
North Am. Food Anim. Pract. 9:309-322.


34
(2003b) found that cows with clinical ketosis were ~ 50 times more likely to develop LDA
than cows without ketosis. However this is not a cause-effect relationship. Ketosis is a
typical metabolic disease explained by low DMI, low levels of glucose, negative energy
balance and a high level of fat mobilization in early postpartum cow (Herdt and Gerloff,
1999; Herdt 2000; Risco and Melendez, 2002). Cows with low DMI at day 1 prepartum
had reduced DMI at day 21 postpartum (Grummer, 1995). Body condition score at calving
is another factor related with DMI and health status of the dairy cows. Cows with excess
BCS at parturition are at increased risk to hypocalcemia, ketosis and LDA (Cameron et ah,
1998; Heuer et ah, 1999). In addition, cows with longer and more extreme period of
negative energy balance have increased risk of digestive problems, including LDA and
laminitis (Collard at ah, 2000).
The review of Shaver (1997) establishes that there are no conclusive studies
reporting that increasing concentrate intake during the last two to three weeks prepartum
reduces postpartum disorders. Curtis et ah (1985), reported that cows with increased
concentrate consumption during the dry period were at lower risk of LDA (OR=0.3 to 0.4)
and ketosis (OR=0.2 to 0.8), respectively. However, Correa et ah (1990), reported that
cows exposed to diets in concentrates were at increased risk of LDA (OR=4.4) and MF
(OR=2.4). Shaver (1997) recommends that until more data are available, an adequate
prepartum concentrate level should be 0.5% of body weight with an upper limit of 0.75%
of body weight. Pre-calving rations containing energy densities > 1.65 Meal of NEL/kg of
DM are associated positively with risk of LDA. These diets are richer in concentrate than
diets < 1.65 Meal of NEL/kg of DM (Cameron et ah, 1998). High concentrate diets, rapid
introduction of concentrate in the immediate pre or postpartum period, rations high in com
silage or low in cmde fiber are factors that affect abomasal motility or enhance gas


123
Results and Discussion
Rumen Metabolites
In Table 4.3 and 4.4 least squares means and standard error of means (SEM) for rumen
metabolites at 10 d pp for primiparous and multiparous cows in treated and control groups are
shown. Primiparous control cows had a tendency to have a lower rumen pH at 4 h after feeding
than cows treated with monensin (Table 4.3). This might be explained because monensin
prevents lactic acid build-up in the rumen and indirectly prevents rumen acidosis (Nagaraja et
al., 1997). Although D and L-lactate were numerically dissimilar in the present study, they
were statistically no different between groups, and comparable with the results obtained by
Mutsvangwa et al. (2002). Although the rest of rumen variables were not statistically different
between groups, numerically, treated cows had lower levels of both acetate and propionate, but
the acetate to propionate ratio was lower in primiparous and higher in multiparous than in
control cows. The lack of significance for VFA might be attributable to either no effect of
treatment or the sample size relative to rumen metabolites may have reduced the power to
detect differences due to treatments (power = 60% or 18 animals per group). The number of
animals per group was calculated expecting an increase in rumen molar proportion of
propionate from 19 to 22 mM 1.2 (Ramanzin et al., 1997; Ruiz et al., 2001).
Multiparous cows treated with monensin had a tendency to have lower ammonia
concentration at 4 h after feeding (P <0.15) and had a significant reduction in rumen ammonia
at 6 h after feeding (P < 0.05). These findings are consistent with previous research, in that
monensin caused a decrease in ammonia production, probably as a result of reduced
proteolysis, degradation of peptides and deamination of amino acids in the rumen (Yang and
Russell, 1993; Nagaraja et al., 1997). Ammonia production by mixed ruminal bacteria under


105
Table 3-1. Diet composition of dry cow far-off, dry cow transition and lactating
Transition cows
Feed (% of diet)
Dry cows
far-off
Dry-cows
transition
Lactating
transition
Alafalfa hay
-
19.3
17.53
Coastal hay
-
4.76
-
Cottonseed whole
-
9.84
8.75
Com silage
44.90
24.06
28.50
Ryegrass silage
15.29
-
4.20
Com hominy
-
6.98
13.54
Citrus pulp
21.20
9.82
4.25
Soybean meal 48
17.20
3.44
5.49
Wet brewers grain
-
10.74
4.07
Lactating concentrate
-
-
7.03
Lactowhey 1
-
3.76
4.23
Metaxerol2
-
2.79
1.96
Springer minerals
-
4.51
-
Dry cow minerals
1.41
-
-
Lactating minerals
-
-
0.45
1 Ammoniated whey (61.5% CP)
2 Energy supplement based on sodium propionate, propylene glycol, dried whey
and calcium carbonate


56
Table 2-5. Ketone body concentrations for clinical and subclinical ketosis according to
different authors
Reference
Class
Ketone body and level
Sauer et al., 1989
subclinical
blood,ketones >0.9 mmol/L
Anderson et al., 1991
hyperketonemic
milk, Ac > 0.4 mmol/L
Gustafsson et al., 1993
hyperketonemic
milk, Ac > 2.0 mmol/L
Gustafsson
and Emanuelson, 1996
hyperketonemic
milk, Ac > 1.4 mmol/L
Shpigel et al., 1996
clinical
urine, AcAc > 60 mg/dL
Duffield,et al., 1998a
subclinical
serum, BHB >1.2 mmol/L
Geishauser et al., 1998
subclinical
serum BHB > 1.2 mmol/L
Jorritsma et al., 1998
subclinical
milk, BHB 1.0 mmol/L
Al-Rawashdeh, 1999
mild ketonemia
severe ketonemia
serum, BHB 0.9-1.7 mmol/L
serum, BHB >1.7 mmol/L
Green et al., 1999
subclinical
serum BHB >1.2 mmol/L
Herdt and Gerloff, 1999
clinical
serum, ketones >35 mg/dL
serum, BHB > 25 mg/dL
Geishauser et al., 2000
subclinical
seru, BHB >1.4 mmol/L
Heuer et al., 2001
subclinical
milk Acetone > 0.4 mmol/L
Melendez et al., 2002a
hyperketonemia
plasma, BHB >10 mg/dL
Reist et al., 2003
subclinical
milk, Acetone > 0.4 mmol/L
serum, BHB > 2.3 mmol/L
Ac= Acetone
AcAc= acetoacetate
BHB= p-hydroxy butyrate
Ketones= total ketone bodies (Ac, AcAc, BHB)


171
Bremmer, D.R., S.J. Bertics, S.A. Besong, and R.R. Grummer. 2000. Changes in hepatic
microsomal triglyceride transfer protein and triglyceride in periparturient dairy cattle J Dairy
Sci. 83:2252-2260.
Bruun, J., A. K. Ersboll, L. Alban. 2002. Risk factors for metritis in Danish dairy cows Prev
Vet. Med. 54:179-190.
Burton, M. J., R. C. Herschler, and H. E. Dziuk.1987. Effect of fenprostalene on postpartum
myometrial activity in dairy cows with normal or delayed placental expulsion. Br Vet J
143:549-554.
Busato, A., D. Faissler, U. Kiipfer, and J.W. Blum. 2002. Body condition scores in dairy
cows: associations with metabolic and endocrine changes in healthy dairy cows. J. Vet. Med.
A 49:455-460.
Butler, W.R., and R. D. Smith. 1989. Interrelationship between energy balance and postpartum
reproduction function in cattle. J. Dairy Sci. 72:767-783.
Cai, T. Q., P. G. Weston, L. A. Lund, B. Brodie, D. J. McKenna, and W. C. Wagner. 1994.
Association between neutrophil functions and periparturient disorders in cows. Am. J. Vet.
Res. 55:934-943.
Callaway, T. R. and S. A. Martin. 1997. Effects of cellobiose and monensin on in vitro
fermentation of organic acids by mixed ruminal bacteria. J. Dairy Sci. 80:1126-1135.
Cameron, R. E. B., R. B. Dyk, T. H. Herdt, J. B. Kaneene, R. Miller, H. F. Bucholtz, J. S.
Liesman, M. J. Vandehaar, and R. S. Emery. 1998. Dry cow diet, management and energy
balance as risk factors for displaced abomasum in high producing dairy herds. J. Dairy Sci.
81:132-139.
Casse, E. A., H. Rulquin, and G. B. Huntington. 1994. Effect of mesenteric vein infusion of
propionate on splanchnic metabolism in primiparous Holstein cows. J. Dairy Sci. 77:3296-
3303.
Chassagne, M., J. Bamouin, and J.P. Chacomac. 1999. Risk factors for stillbirth in Holstein
heifers under field conditions in France: a prospective survey. Theriogenology 51:1477-1488.
Christensen J. O., R. R. Grummer, F. E. Rasmussen, and S. J. Bertics. 1997. Effect of method
of delivery of propylene glycol on plasma metabolites of feed-restricted cattle. J. Dairy Sci.
80:563-568.
Clanton, D.C.; M.E. England; J.C. Parrott. 1981. Effect of monesin on efficiency on
production in beef cows. J. Anim. Sci. 53:873-880.
Collard, B. L., P. J. Boettcher, J. C. M. Dekkers, D. Petitclerc, and L. R. Schaeffer. 2000.
Relationships between energy balance and health traits of dairy cattle in early lactation. J.
Dairy Sci. 83:2683-2690.


Sensitivity
168
1-Specificity
Figure 6-1. ROC curve for milk BHB for the detection of subclinical
ketosis using a gold standard of serum BHB > 1.0
mmol/L


161
of serum BHB of 1.0 mmol/L and a milk BHB cut-off value of 200 pmol/L will be used for
further discussion of the results.
Twenty cows were culled between assignment and 14 d postpartum. Therefore, only
280 cows were tested for BHB in milk. Overall, the incidence of subclinical ketosis based
on the milk test was 20.35% (57/280). Other studies using the same ketone test have shown
incidence values below 14% (Jorritsma et al, 1998; Geishauser et al., 2000). Based on milk
BHB test, 26.6% of control cows (n=139) and 14.5% of treated cows (n=141) were positive
for subclinical ketosis. The adjusted effect of treatment was that cows receiving monensin
were 0.68 times less likely to give a result of 200pmol/L (OR 95% Cl =0.53-0.80) than
control cows (Table 6.5).
Concentrations of BHB in serum did not differ between groups (0.65 0.07 mmol/L
and 0.70 0.07 mmol/L for controls and treated, respectively) (P > 0.05), however cows
with serum BHB concentration < 1.0 mmol/L were 0.063 times (Cl 95% 0.013-0.302) less
likely to have a positive result for milk BHB (200 pmol/L). These results are in agreement
with other studies where high levels of BHB in blood were correlated with positive results
of BHB in milk (Jorritsma et al., 1998; Geishauser et al., 2000). In the present study,
concentration of BHB in serum was lower than in other studies that used monensin as a
controlled-release capsule, with values ranging between 0.83 and 1.1 mmol/L at 2 weeks
post partum (Green, et al., 1999; Duffield et al., 2003). These values agreed with blood
BHB concentrations reported in a previous study conducted on the same farm in 1998
(Melendez et al., 2002a).
The present study demonstrated that cows fed diets containing citrus pulp and
supplemented with monensin through a slow-release capsule during the transition period
experienced a decrease incidence of subclinical ketosis at 14 d postpartum. Although other


129
Table 4-3. Least squares means S.E.M. for rumen pH, acetate, propionate, butyrate,
valerate, total VFA, ammonia, D and L-lactate and acetate to propionate ratio
by time in primiparous treated and control cows at 10 d pp
PH
Ac1
Pr2
Bu3
Va4
VFA5
NHj
L-Lacr
D-Lac7
A/P8
mMol/L
M9
6.89
49.9
19.5
10.1
0.92
83.8
2.32
0.02
0.03
2.56
TO
SEM
0.19
8.05
4.8
1.55
0.22
14.7
0.77
0.18
0.28
0.22
c'
6.82
58.9
22.6
10.2
1.06
96.1
1.71
0.05
0.03
2.73
SEM"
0.10
4.30
2.6
0.83
0.12
7.87
0.41
0.10
0.15
0.11
M
6.53
64.8
30.1
13.1
1.64
114.7
5.59
0.04
0.01
2.09
T2
SEM
0.19
8.05
4.8
1.55
0.22
14.7
0.77
0.18
0.28
0.22
C
6.49
68.2
31.8
12.7
1.66
118.1
4.69
0.23
0.36
2.16
SEM
0.10
4.30
2.6
0.83
0.12
7.87
0.41
0.10
0.15
0.11
T4
M
6.62*
65.1
29.1
13.7
1.60
113.5
4.59
0.04
0.13
2.24
SEM
0.19
8.05
4.8
1.55
0.22
14.7
0.77
0.18
0.28
0.22
C
6.34*
70.9
31.1
13.1
1.67
120.6
4.42
0.07
0.09
2.31
SEM
0.10
4.30
2.6
0.83
0.12
7.87
0.41
0.10
0.15
0.12
T6
M
6.49
63.3
26.6
13.8
1.47
109.2
3.73
0.03
0.01
2.38
SEM
0.19
8.05
4.8
1.55
0.22
14.7
0.77
0.18
0.28
0.22
C
6.32
69.6
29.4
12.8
1.50
116.9
3.51
0.06
0.05
2.42
SEM
0.10
4.30
2.6
0.83
0.12
7.87
0.41
0.10
0.15
0.11
* (P < 0.15)
1 Acetate,2 Propionate,3 Butyrate, 4 Valerate,5 Total Volatile Fatty Acids,6 L-Lactate,7 D-
Lactate,8 Acetate to Propionate Ratio, 9 Monensin,10 Control,11 Standard Error of Mean, TO
time 0, 7:00 A.M. before feeding; T2= time 2, 2 h after A.M. feeding; T4= time 4, 4 h after
A.M. feeding; T6= time 6, 6 h after A.M. feeding.


35
production (Nocek et al., 1983; Markusfeld, 1986; Trent, 1990). Dawson et al. (1992),
reported that cows fed ground alfalfa hay and concentrate in a TMR in early postpartum
were at higher risk for LDA than were cows fed the standard herd ration of sorghum silage
and concentrate mixed plus loose alfalfa hay. The lack of physical form reduces chewing
activity, ruminal fill, motility and fiber mat formation and increases ruminal VFA
concentration, all of which may affect the etiology of LDA. Concentrate DM can be
increased at the rate of 0.20 to 0.25 kg/d until peak lactation is reached. Concentrates
should be fed three to four times daily. Feeding a TMR to control forage:concentrate ratio
is recommended. A transition group TMR with a higher effective fiber content for early
postpartum cows is also recommended. The importance of physical form of fiber as a risk
factor for LDA is likely to be greatest during the early postpartum period because of the
physiologic and metabolic changes in the transition period. The lack of physical form
reduces chewing activity, ruminal fill, motility, fiber mat formation, and increases ruminal
VFA concentration, all of which may affect the etiology of LDA (Shaver et al., 1986;
Bauchemin, 1991; Mertens, 1992; Muller, 1992; Shaver, 1997; Varga et al., 1998;
Heinrichs et al., 1999).
Different forage program for dry cows are used under field conditions, but data are
limited regarding their impact on the incidence of LDA (Shaver, 1997). Nocek et al.
(1983) evaluated three dry cow forage programs consisting of long hay, hay and com
silage (50% each one) and com silage. The incidence risk for LDA was 3.0, 4.3 and 6.3%,
respectively. The incidence risk for ketosis was 9.1%, 6.3 and 6.4%, respectively. Shaver
(1997) recommends that rations with 100% com silage as forage source should not be fed
to dry cows. The controlled use of com silage as a component of forage programs for dry
cows may be beneficial.


85
Vitamin levels should be met to optimize milk production. Vitamin A should be
provided at rate of 3600 IU per kg DM/day. Vitamin D 900 IU per kg/DM day and
vitamin E 14 IU per kg/DM day (Mahanna, 1997).
Drv Cow Feed Management and Body Condition Score
Body condition scoring is a useful tool for monitoring the nutritional management
of dairy cows (energy density and intake). Using a scale 1 to 5 (Edmonson et al., 1989;
Ferguson et ah, 1994) a program can be established. Beede (1997) recommends that, at
dry-off, cows should have a BCS 3.0 to 3.25. If BCS is lower, the ration should be
adjusted during the last 100 days of lactation and not during the dry period. If many cows
are over-conditioned, a fat cow group for late lactation should be established and a diet
containing 1.54 Mcal/Kg NE1/DM or less should be fed. If many cows are
underconditioned a thin cow group should be established and a diet with proper NE1
content to target the desired weight gain should be fed (one unit BCS 57 kg BW).
Cows should have a total dry period length of at least 6 to 8 weeks. A dry period of
less than 6 weeks results in lower milk yields in the next lactation (Funk et ah, 1987)
Two groups of dry cows are recommended; the early dry group (8 to 3 weeks
before expected calving) and a close-up group (3 weeks to calving). If early dry cows are
in proper condition and eating more than predicted to meet their requirements, it may be
necessary to lower the energy and nutrient density of the ration by re-formulating the diet.
Never try to reduce BCS or body weight of dry cows during any stage of the dry period (
Beede, 1997). If BCS is correct at dry-off (3.0-3.25) then cows should gain about 0.3-0.45
kg/d to increase BCS by 0.25 to 0.35 units during the early period.
In the close-up period, if DMI is around 11.3 kg or higher, a diet containing 1.6
Mcal/kg NE1 but no higher, and 14-15% of Crude Protein should be fed (NRC, 2001). A


160
Pi = parameter of X]
Xi = effect of treatment
P2 = parameter of X2
X2 = effect of parity
p3 = parameter of X3
X3 = effect of milk yield at 14 days postpartum
P4 = parameter of X4
X4 = effect of body condition score at calving
Results and Discussion
At assignment, groups did not differ in parity, BCS and milk yield of previous lactation
(P > 0.05).
Based on three concentrations of BHB in serum (> 0.8 mmol/L; > 1.0 mmol/L and > 1.2
mmol/L) a ROC analysis was conducted (Table 6.3). The highest agreement between tests
(kappa estimator = 0.37) occurred at the value of 1.0 mmol/L. Therefore, the value of 1.0
mmol/L was used as the gold standard to classify an animal as positive or negative for
subclinical ketosis. In Figure 6.1, ROC analysis for milk BHB based on a gold standard of
serum BHB >1.0 mmol/L is shown. In Table 6.4, sensitivity, specificity, positive and negative
predictive values for different concentration values of milk BHB test at a level of serum BHB >
1.0 mmol/L (gold standard) are shown. Based on the ROC curve, the value of milk BHB of 200
pmol/L gives the best combination of sensitivity and specificity to classify an animal as positive
or negative (Se=52.9; Sp=86.4). This is in accordance with the recommendations of the
commercial test strip used in the present study. However, Se was 6.01% lower and Sp was
3.65% lower than values reported by Gaishauser et al. (2000) (Se=59%; Sp=90). In another
report, using the same milk ketone test with a cut-off of 200 pmol/L, Se was 40% and Sp was
94% (Jorritsma et al., 1998). For the purpose of this study a gold standard cut-off concentration


To Oscar and Eliana, my parents,
I wish to dedicate this dissertation to those who guided my life. They have
been suffering for being far away from my kids, my wife and my-self. Tears, efforts,
and no resting have characterized their lives; but they should be very proud for having
a son who at this moment has become faculty of one of the most prestigious Colleges
of Veterinary Medicine of the USA.
Dad and Mom, this is not mine, this is yours. Without you I would have been
nothing. I love you so much and please pardon me for not being with you.


12
Table 2-3 Continued
Author
Disease
Risk factors
Association
Grohn et
Ketosis
Parity
Positive
al., 1989
Milk yield
Positive
continued
Milk fever
OR= 1.6
LDA
OR= 2.5
Metritis
OR= 2.3
Mastitis
OR= 1.4
Lameness
OR= 2.4
Correa et
Milk fever
Lead feeding
OR= 2.4
al., 1990
RFM
Low Ca diets
OR= 1.7
Farmer treatments
OR= 1.7
Metritis
Dystocia
OR= 5.6
RFM
OR= 86.5
LDA
Metritis
OR= 43.7
Leading feeding
OR= 4.4
Ketosis
Milk fever
OR= 41.5
Dry fat cows
OR= 3.1
Mastitis
RFM
OR= 8.7
Milk fever
OR= 31.3
Correa et
RFM
Dystocia
OR= 2.2
al., 1993
Twinning
OR= 3.4
Metritis
Dystocia
OR= 2.1
RFM
OR= 6.0
Ketosis
OR= 1.7
Ketosis
Milk fever
OR= 2.4
LDA
Milk fever
OR= 2.3
Ketosis
OR= 13.8
Dystocia
OR= 2.3
Dystocia
Twinning
OR= 10.5
Milk fever
OR= 2.6


112
Days post partum
Figure 3-3. Daily milk yield up-to 20 d pp in third lactation cows or older by treatment


18
Stage I may be treated with either oral calcium supplements or intravenous
calcium salts. Animals in stage II or III require immediate treatment with intravenous
calcium salts (Oetzel and Goff, 1999). The fastest way to restore normal plasma Ca
concentration is to administer an intravenous injection of Ca salts (Goff, 1999a). Calcium
gluconate or borogluconate is the standard intravenous treatment in cows. Five hundred ml
of these products provide 10.8 g of calcium. Commercial preparations usually supply
between 8.5 to 11.5 g Ca/500 ml. They may also contain Mg, phosphite and glucose. All
the preparations effectively raise total and ionized Ca concentrations in blood (Goff,
1999a; Oetzel and Goff, 1999). Intravenous Ca should always be administered slowly to
prevent sudden cardiac arrest due to hypercalcemia (Oetzel and Goff, 1999). Calcium
should be administered at a rate of 1 g/minute (Goff, 1999a). Approximately 60% of
recumbent animals affected with uncomplicated MF will get up within 30 minutes after a
single intravenous dose with calcium salts. Another 15% can be expected to rise within the
next 2 hours. Full restoration of normal calcium homeostasis usually requires 2 or 3 days.
About 10% of dairy cows with MF stay recumbent for over 24 hours but eventually
recover (Oetzel and Goff, 1999).
Ca salts can also be injected subcutaneously. The serum Ca concentration achieved
is not the same as with intravenous administration with similar dose. However, as with
intravenous Ca injection, the subcutaneous dose of Ca increases Ca in blood for 4 to 5
hours only (Goff, 1999a). Greater amount of Ca can cause local tissue necrosis; then Ca
injections should be limited to 1 to 1.5 g Ca (50 to 75 mL of most commercial
preparations) per site. Calcium chloride solutions are not well tolerated subcutaneously.
Ca solutions containing glucose may also be slightly more injurious. Preparations are
available for intramuscular administration of Ca in the form of calcium levulinate or


120
Experimental Protocol
In March, 2003, 24 cows dried-off 50 to 70 days BEP were randomly assigned either a
treatment or a control group. The treated group (n=12) received orally a capsule of monensin
(releasing 300 mg of monesin daily for 95 days, CRC Rumensin, ELANCO Animal Health,
Guelph, ON, Canada). Control cows (no capsule, n=12) were randomly matched by parity.
The number of animals per treatment was calculated expecting an increase in rumen
molar proportion of propionate from 19 to 22 mM (SEM 1.2) (95% confidence, 80% of
power).
Treated and control cows received the same diets (Tables 4.1 and 4.2), were exposed to
the same environment and were handled homogeneously during the entire experimental
protocol (far-off and transition period).
Cows were monitored for health problems during the entire experimental period and
pens were periodically inspected to observe if boluses were regurgitated. No boluses were
recorded to be lost.
Blood Sampling and Body Condition Score
At assignment, on day 21 BEP, at calving, and at 7, 14, and 21 d postpartum a blood
sample was obtained and a body condition score (BCS) evaluation was conducted, at the same
time of the day by the same person. Blood samples were collected from the coccygeal vein,
using an evacuated tube without anticoagulant for serum collection, heparinized tubes for
plasma metabolite analysis and in NaFl tubes for glucose analysis (Vacutainer, Becton
Dickinson, Rutherford, NJ). Body condition score was evaluated at assignment and at calving
using a scale of 1 to 5 based on standard methodology (Ferguson et al., 1994)


90
Table 2-11. Target Body Condition Scores (BCS) Scale 1-5
Stage
Ideal bcs
Range
Dry-off
3.5
3.25-3.75
Calving
3.5
3.25-3.75
Early lactation
3
2.5-3.25
Mid-lactation
3.25
2.75-3.25
Late lactation
3.5
3.0-3.5
Growing Heifers
3
2.75-3.25
Heifers at calving
3.5
3.25-3.75
- .i-,th
Adapted from: National Research Council. 2001. Nutrient requirements of dairy cattle, 7th
revised edition. Washington, D.C.: National Academy Press.


159
treatment groups. Serum BHB concentration was analyzed by ANOVA, mixed model.
Proportion of cows positive to the milk BHB test was analyzed by logistic regression.
Sensitivity, specificity, positive and negative predictive value for milk ketone tests were
calculated. A ROC (receiver-operating characteristic) analysis for BHB in serum and ketone
levels in milk was conducted (Greiner et al., 2000). A ROC curve is a plot of the sensitivity
of a test versus the false positive rate (1-specificity) computed at a number of different cut
off values to select the optimum cut-off value for distinguishing between positive and
negative animals. The closer the ROC curve reaches to the top-left comer of the graph, the
better the ability of the test to discriminate between positive and negative animals. The very
top-left comer represents a test with a sensitivity of 100% and a specificity of 100% as well
(Dohoo et al., 2003). Agreement evaluation between BHB in blood and BHB in milk was
performed by the kappa estimator. Variables were considered significant at P < 0.05.
Statistical analysis was conducted using SAS 8.2 (SAS, 2001) and Winepiscope (2001).
Mixed model was defined as:
y¡ji= p + Tj + + Pap + (Par *T)n + eyi
Where:
y¡ji= BHB concentrations
T¡= fixed effect of treatment
Cow (Tj)j = random effect of cow nested in treatment
Par i = fixed effect of parity
(Par *T) ¡i = interaction parity and treatment
e¡ji = random error term
Logistic model was defined as:
Logit (ti) = a + P,X, + p2X2 + p3X3 + P4X4
Where:
7i = log of the odds of the event (subclinical ketosis yes, no)
a = intercept


subsample of 50 cows per group, a blood sample for BHB determination was taken. In
study 4, 24 cows (in March 2003) were assigned similarly. A blood sample was obtained
and BCS was conducted at assignment; on day 21 BEO; at calving; and at 7, 14, and 21 d
postpartum. A rumen fluid and blood sampling scheme was carried out on day 10
postpartum. The first sample was obtained before the morning meal. Three more samples
were taken every 2 hours. Outcome variables in rumen samples were pH; and
concentrations of NH3, acetic, propionic, butyric, and L and D-lactic acids. Outcome
variables in blood samples for studies 2 and 4 were non-esterified fatty acids (NEFA),
BHB, and glucose.
In study 1, monensin improved milk yield, decreased the incidence of metritis and
increased the incidence of dystocia in third and older parity. Milk solids were decreased
by treatment. In study 2, monensin increased BCS at calving in multiparous cows. In
study 3, monensin decreased the proportion of cows with subclinical ketosis. In study 4,
monensin decreased the level of BHB and NEFA; and increased glucose levels. Minor
changes in rumen fermentation were detected. It is concluded that monensin improved
slightly milk production and metabolism performance of transition dairy cows fed diets
containing citrus pulp.
xiii


134
Table 4-8. Least squares means S.E.M. for serum b-hydroxy butyrate, non-esterified fatty
acids and glucose by time of sampling in multiparous treated and control cows
between dry-off and 21 d pp
Time
Group
BHB
NEFA
Glucose
mmol/L
meq/L
ing/dL
0
M
0.32 0.11
0.24 0.12
60.02 4.31
C
0.27 0.17
0.30 0.19
67.22 6.47
1
M
0.44 0.11
1.20 0.12
72.63 4.87
C
0.48 0.17
1.00 0.19
71.24 7.44
2
M
0.76 0.11
0.89 0.12
57.82 4.09
C
0.83 0.17
0.89 0.19
55.74 5.79
3
M
0.77 0.11
0.89 0.12
55.80 5.26
C
0.85 0.17
0.85 0.19
59.66 6.46
4
M
0.79 0.11
0.90 0.12
57.35 4.31
C
0.76 0.17
0.98 0.19
55.42 6.46
5
M
0.70 0.11
0.74 0.12
55.23 5.27
C
0.93 0.17
0.75 0.17
60.23 5.28
0: assignment, dry-off; 1: 21 d before expected parturition; 2: Calving; 3: 7 d pp; 4: 14 d pp 5
21 d pp


23
ingredients as a premix could offer an inexpensive, safe and palatable alternative to
anionic salts (Goff and Horst, 1998b). Optimal acidification generally occurs when anions
are added to achieve a final DCAD (using equation 1) between -50 to -150 mEq/kg of dry
matter. The strong negative relationship (r2=0.95) between urinary pH and net acid
excretion by cows fed the diets containing anionic salts suggests that urinary pH
measurement might be a useful tool to assess the degree of metabolic acidosis that is
imposed by dietary anionic salts (Vagnoni and Oetzel, 1998). An advantage of this
approach is that it accounts for inaccuracies in mineral analyses and for unexpected
changes in forage mineral content. Urinary pH can be evaluated by obtaining urine from a
group representing about 10% of the pre-calving cows. Urinary pH values below 5.5
indicate overacidification and DCAD should be increased. The optimal urinary pH is
between 6.0 and 6.5 for Holstein cows and between 5.8 and 6.2 for Jersey cows. Over 6.5
is considered inadequate acidification and suggests that a lower DCAD is required. In
herds experiencing MF the urine of close-up dry cows will be very alkaline with a pH
above 8.0. Most accurate results will be obtained by collecting urine samples at a standard
time, preferably within a few hours after feeding (Goff and Horst, 1998a; Oetzel and Goff,
1999).
Retained Fetal Membranes Metritis Complex
Retained Fetal Membranes (RFM) is defined as the lack of detachment of fetal
membranes from the maternal caruncles within the first 12 to 24 hours after calving
(Grunert, 1986; Eiler, 1997). Van Werven et al., (1992) reported that 77.3% of cows
expel the fetal membranes by 6 hours. Normal expulsion of fetal membranes requires that
the maternal and fetal tissues undergo maturation and a loosening process, which is
completed by 2 to 5 days before the end of an average gestation. These changes include


46
The effect of ketosis on culling was determined described in Holstein cattle in New
York State by two groups of researchers in two different time periods (Milian-Suazo et al.,
1988; Grohn et al., 1998) They found that ketotic cows were more likely to be culled
throughout lactation than were nonketotic cows. However, Gillun et al. (2001) in Norway
found no difference in culling rate between ketotic and nonketotic cows.
During early lactation dairy cows experience a typical negative energy balance
characterized by mobilization of NEFA from adipose tissue (Goff and Horst, 1997b; Herdt
and Gerloff, 1999; Herdt, 2000). This is explained by low DMI at parturition and slower
increase in DMI than in milk production during the early postpartum period. Energy
required for maintenance and milk production exceeds the amount of energy the cow can
obtain from dietary sources (Goff and Horst, 1997b).
Herdt and Gerloff, (1999) establish that distinct metabolic types of ketosis might
exist dependent upon hepatic patterns of NEFA metabolism. Glucose availability is an
important factor in the pathogenesis of clinical ketosis. When glucose availability is very
low, entry of NEFA into a ketogenic liver pathway is favored, but when glucose is high,
esterification and fat deposition are favored (Herdt, 2000). Two factors determine glucose
availability: (1) rate of gluconeogenesis and (2) availability of gluconeogenic substrate.
Rate of gluconeogenesis is increased by the abundance of pyruvate carboxylase during the
early transition period followed by increased abundance of phosphoenol pyruvate
carboxykinase (PEPCK) during the postpartum period (Greenfield et al., 2000). Rate of
gluconeogenesis may be impaired in those cases of ketosis that develop within the first
week postpartum. Evidence suggests that hepatic fat accumulation prior to calving may
interfere directly or indirectly with gluconeogenesis. It has been reported that activity of
PEPCK is decreased in cows that developed fatty liver postpartum (Rukkwamsuk et al.,


166
Table 6-4. Sensitivity, specificity, positive predictive value, negative predictive value of
milk BHB test based on a serum BHB gold standard cut-off value of 1.0
mmol/L
Milk BHBA
concentration
pmol/L
Sensitivity
Specificity
Positive
predictive value
Negative
predictive value
50
100
20.9
20.9
100
100
94.1
67.9
38.1
98.2
200
52.9
86.4
45
89.7


184
Nocek, J. E., J. E. English, and D. G. Braund. 1983. Effects of various forages feeding
programs during dry period on body condition score and subsequent lactation health,
production and reproduction. J. Dairy Sci. 66:1108-1118.
Oetzel, G. R. 1988. Parturient paresis and hypocalcemia in ruminant livestock. Vet. Clin.
North Am. Food Anim. Pract. 4:351-364.
Oetzel, G. R. 1998. Nutritional management of dry dairy cows. Compend. Contin. Educ. Prac.
Vet. 20:391-396.
Oetzel, G.R. 2000. Management of dry cow for the prevention of milk fever and other mineral
disorders. Vet. Clin. North Am. Food Anim. Pract. 16:369-386.
Oetzel, G. R., and J. P. Goff. 1999. Milk fever in cows, ewes and doe goats. Pages 215-218 in
Current Veterinary Therapy 4. Food Animal Practice. J. Howard and R. Smith. W.B.
Saunders Company.
Olsen, G.F. 1962. Optimal conditions for the enzymatic determination of L-lactic acid. Clin.
Chem. 8:1-10.
Olson, J. D., K. Bretzlaff, R. G. Moltimer, and L. Ball. 1986. The metritis-pyometra complex.
Pages 227-236 In Current therapy in theriogenology. D.A. Morrow. W.B. Saunders, Co.,
Philadelphia, PA.
Oltenacu, P. A., J. H. Britt, R. K. Braun, and R. W. Mellenberger. 1984. Effect of health status
on culling and reproductive performance of Holstein cows. J. Dairy Sci. 67:1783-1792.
Orpin, C.G., and K.N. Joblin. 1997. The rumen anaerobic fungi. Pages 140-195 in The rumen
microbial ecosystem. Second edition. P.N. Hobson and C.S. Stewart. Blackie Academic &
Professional.
Oscar, T.P. and J.W. Spears. 1990. Incorporation of nickel into ruminal factor F430 as affected
by monensin and formate. J. Anim. Sci. 68:1400-1404.
Ostergaard, S., and Y. T. Grohn.1999. Effects of diseases on test day milk yield and body
weight of dairy cows from Danish research herds. J. Dairy Sci., 82:1188-1201.
Overton, T.R., J.K. Drackley, G.N. Douglas, L.S. Emmert, and J.H. Clark. 1998. Hepatic
gluconeogenesis and whole-body protein metabolism of periparturient dairy cows as affected
by source of energy and intake of the prepartum diet. J. Dairy Sci. 81(Suppl. 1):295. (Abstr.)
Overton, T.R., J.K. Drackley, C.J. Otteman-Abbamonte, A.D. Beaulieu, L.S. Emmert, and
J.H. Clark. 1999. Substrate utilization for hepatic gluconeogenesis is altered by increased
glucose demand in ruminants. J. Anim. Sci. 77:1940-1951.


125
In general, BHB and NEFA concentrations were lower and glucose concentrations were
higher in the monensin group than control group, both at 10 d pp and during the entire
experimental period between dry-off and 21 d pp, in primiparous and multiparous cows. These
results were expected and corroborate previous findings from other studies that used monensin
in a controlled-release capsule with typical diets high in concentrates (Duffield et al., 1998a;
1998b; Green et ah, 1999; Duffield et ah, 2003) and cows fed on pasture and supplemented
with concentrates (Abe et ah, 1994; Stephenson et ah, 1997). Conversely, Hayes et ah (1996)
did not find a positive effect of intraruminal monensin capsules on BHB, NEFA and glucose;
however in that particular study, cows were fed only on pasture with a high content in crude
protein (16.3 20.5% DM basis) and no concentrate supplementation. Interestingly,
independent of treatment, NEFA, BHB and glucose increased dramatically at the close-up
period, indicating that cows around three weeks prepartum started to change their metabolic
status, a condition that has been previously reported (Grum et ah, 1996; Herdt, 2000; Drackely,
2001). During the first week postpartum NEFA and BHB were noticeably lower in treated than
control primiparous cows. This difference was not evident in multiparous cows. Also,
differences in hourly metabolite concentrations at 10 d pp were more evident in primiparous
than multiparous cows. These patterns clearly suggest that monensin had a more positive effect
in first lactation animals than mature cows. This observation might be apparent since first
lactation cows have lower milk production than adult animals. By producing more milk, older
cows would experience a higher metabolic status. This condition would worsen if adult cows
treated with monensin were higher producers than adult cows not treated with monensin.
Under this scenario, metabolic differences should be minimal.


49
liver could impact regulation of NEFA and glucose metabolism in the liver (Drackley et
al., 2001). In cows with fatty liver, content of C 18:1 increased and C 18:2 decreased
relative to prepartum values. In addition, elevated concentration of circulating NEFA is
associated with increased liver concentrations of palmitic, oleic, and linoleic acids, but not
stearic acid (Rukkwamsuk et al., 1999c; 2000).
Methods of diagnosis of ketosis-fatty-liver complex deserve attention. Ketosis is
usually diagnosed based on clinical sings and the level of ketone bodies in urine or milk.
Subclinical ketosis is determined not simply by the presence of ketone bodies, but an
abnormal level of circulating ketone bodies (Duffield, 2000). Ketone bodies can be
measured in blood, urine and milk. In Table 2-5 ketone body concentration from different
studies are reported.
When ketone bodies are measured quantitatively, a defined threshold must be used
to separate normal animals from those with subclinical ketosis (Duffield, 2000). Several
field testing kits have been developed with different sensitivity and specificity values. In
Table 2-6 two studies are summarized.
Subclinical ketosis may start at serum BHB concentrations above 1.0 mmol/L
(10.4 mg/dL) and clinical ketosis at 2.6 mmol/L (27 mg/dL), however these levels are
extremely variable among animals (Duffield, 2000). Serum concentration of 1.4 mmol/L
of BHB (15 mg/dL) or greater in the first 2 weeks postpartum was found to cause a three
fold greater risk for cows to subsequently develop either clinical ketosis or abomasal
displacement. In addition, cows having serum BHB concentrations at or above 2.0
mmol/L (21 mg/dL) within 7 days prior to the first Dairy Herd Improvement (DHI) test
produced over 4 kg less milk on DHI test day (Duffield, 1997). Despite its instability,
blood acetoacetate levels have been used by some authors to identify animals with


131
Table 4-5. Least squares means S.E.M. for serum 13-hydroxy butyrate, non-
esterified fatty acids and glucose by time of sampling after A.M.
feeding in primiparous treated and control cows at 10 d pp
Time
Group
BHB
mmol/L
NEFA
mEq/L
Glucose
mg/dL
TO
M
0.81 0.22
0.82 0.18n
52.9 4.90
C
0.86 0.12
1.14 0.10t+
48.3 2.60
T2
M
0.65 0.22
0.60 0.18
54.4 4.90n
C
1.00 0.12
0.47 0.10
45.9 2.60n
T4
M
0.57 0.22+
0.43 0.18
54.2 4.90
C
1.000.12t
0.59 0.10
48.5 2.60
T6
M
0.71 0.22n
0.51 0.18
51.6 4.90
C
0.99 0.12+t
0.60 0.10
48.3 2.60
TT (P<0.1)
f (p < 0.05)
TO: at A.M. feeding; T2: 2 h post feeding; T4: 4 h post feeding; T6: 6 h post feeding


14
Table 2-4. Case definition, incidence and economic losses of calving-related disorders
Disease
Case definition
Incidence
Economic losses
Milk fever
Calcium deficiency causing
progressive neuromuscular
dysfunction with flaccid paralysis,
circulatory collapse, and depression of
consciousness
Median 6.5%
Range .03 -
22.3%
$335 per case
RFM
Fetal membranes visible at the vulva
or in vagina or uterus by vaginal
examination more than 24 h after
parturition
Median 8.6%
Range 1.3 -
39.2%
$285 per case
Metritis
Abnormal cervical discharge, vaginal
discharge, or both or uterine content.
New case if cow did not have a case
during the preceding 30 days
Median 10.1%
Range 2.2 -
37.3%
Treatment,
increased days
open and culling
Ketosis
Primary: Decreased appetite, elevated
milk, urine or breath ketones in the
absence of other disease
Median 4.8%
Range 1.3-
18.3%
$145 per case
LDA
Decreased appetite accompanied by an
audible, high pitched tympanic
resonance (ping) by percussion of the
left abdominal wall between the 9th
and 12th ribs
Median 1.7%
Range 0.3 -
6.3%
$340 per case.
Milk losses 250-
2000 kg/
lactation
Ovarian
cysts
Smooth, rounded structure greater
than 25 mm in diameter in one or both
ovaries non pregnant cows
Median 8.0%
Range 1.0 -
16.0%
$39 per case
Lameness
Mastitis
Episode of abnormal gait attributable
to either the foot or leg regardless of
etiology or duration
Visually abnormal milk secretion from
one or more quarters with or without
signs of inflammation of the udder.
New case following 8 days of normal
milk
Median 7.0%
Range 1.8 -30%
Median 14.2%
Range 1.7 -
54.6%
$302 per case
Adapted from: Kelton, D. F., K. D. Lissemore, and R. E. Martin. 1998. Recommendations
for recording and calculating the incidence of selected clinical diseases of dairy cattle. J.
Dairy Sci. 81:2502-2509.


146
Blood Metabolites
Primiparous and multiparous cows, demonstrated a significant pattern over time of the
three metabolites. These patterns are consistent with previous reports where NEFA start to
increase about three weeks prepartum and peak at calving, glucose is high prepartum until
parturition and then declines after calving and BHB starts to increase prepartum reaching a peak
between 7 to 14 d pp (Abe et al., 1994; Stephenson et ah, 1997; Duffield et ah, 1998a; Arielli et
ah, 2001; Vallimont et ah, 2001).
In primiparous cows, neither BHB, glucose nor NEFA differed between groups (P >
0.05). However in multiparous treated cows, glucose was significantly higher than controls at
parturition (P < 0.05). Although in previous studies, monesin improved the metabolic status of
primiparous cows (Green et ah, 1999), in the present study that effect was not found. One
explanation might be that first lactation cows in the present study were higher producers than
heifers in the Canadian study (35.3 kg/d vs 23.5 kg/d, respectively). Therefore, primiparous
cows in the present study were exposed to higher metabolic and productive stresses that may
have masked the effect of monensin based on their higher genetic merit. In addition, prepartum
and postpartum transition diets in the present study were offering other gluconeogenic
compounds (sodium propionate, propylene glycol (Metaxerol, Lactowhey*)) that perhaps
counterbalanced the effect of monensin.
In conclusion, a monensin controlled release capsule inserted at dry off tended to
increase milk production and increased BCS at calving in multiparous cows fed citrus pulp. In
primiparous cows, milk yield and BCS at calving did not differ between groups. Energy related
blood metabolites were not different between groups in both primiparous and multiparous cows.


75
increased production in response to monensin treatment. Cows with a BCS between 3.25
and 3.75 produced significantly more milk at the second DHIA test (0.85 kg), and cows
with a BCS > 4.0 produced significantly more milk than did controls for all three DHIA
tests (1.25 kg/d). Treatment with monensin had no significant effect on either milk fat or
milk protein percentage (Duffield et al., 1999a). After this, Duffield et al., (1999b)
determined the effect of a monensin-controlled release capsule on cow health and
reproductive performance. They found, in 503 cows receiving the bolus, that monensin
significantly reduced the incidence of displacement of the abomasum (OR=0.41-0.84) and
multiple illnesses (OR=0.38-0.89), defined as having more than one of retained placenta,
milk fever, metritis, endometritis, mastitis, ketosis, displaced abomasums, digestive,
lameness, respiratory disease, or other disease. Treatment was not statistically significant
in reducing the incidence of clinical ketosis (P=0.11), defined as reduced feed intake,
testing positive in a milk ketone test, and the absence of any other disease. Treatment with
monensin had no significant effect on the interval from calving to first service. Similarly,
treatment had no effect on the interval calving to conception. Service per conception and
conception rate at first service did not differ between groups as well.
In an effort to complement results on the prevention of subclinical ketosis in
lactating dairy cows, Green et al. (1999) designed an experiment to study subclinical
ketosis in periparturient dairy cows and the antiketogenic effects of monensin. In this
study, the authors induced subclinical ketosis (n=41) through 10% feed restriction and
blood BHB quantification (using a blood BHB threshold of 1200 pmol/L). No treatment
effects were found for daily milk production, milk components, dry matter intake, net
energy balance, body weight and BCS. Rumen pH for the postpartum period was higher
for cows treated with monensin. Cows treated with monensin had lower concentration of


53
decreased as propylene glycol dose was increased, indicating ruminal conversion of
propylene glycol to propionate (Grummer et al., 1994; Christensen et al., 1997). In liver,
propylene glycol is converted to glucose, primarily via the lactaldehyde pathway and
subsequent oxidation to lactate (Nelson and Cox, 2000).
Recommended doses of propylene glycol have been in the range of 250 to 400 g,
administered orally, twice a day (Herdt and Gerloff, 1999). Toxicity has been seen with
higher doses. Neurological symptoms develop in 2 to 4 hours and resolved by 24 hours
after dosing. Ataxia is typical. Depression and recumbence are also experienced. Serum
and cerebrospinal fluid osmolality is increased. The median toxic dose of propylene glycol
in cattle is 2.6 g/kg of body weight (Pintchuk et al., 1993).
Administration of propylene glycol before parturition has demonstrated to decrease
hepatic fat accumulation and ketone bodies formation. One liter of propylene glycol given
orally once daily during the last 10 days prepartum increased plasma glucose in the treated
group before parturition, but glucose levels were similar between treated and control
group, after calving. Plasma NEFA concentration was 403 and 234 pM, and plasma
insulin concentrations were 0.354 and 0.679 ng/ml for control and treated cows,
respectively from 1 to 7 days prepartum. Plasma NEFA tended to be lower in treated cows
at 1 to 21 days post partum. In the treated group, hepatic TG accumulation was reduced 32
and 42% at 1 and 21 days post partum, respectively. Prepartum plasma glucose, NEFA,
BHB and insulin were strongly correlated with liver TG at 1 day postpartum (r = -0.49,
0.45, 0.36, and -0.49, respectively). Milk production and composition through 21 days
postpartum were not different between the treated and control groups (Studer et al., 1993).
Grummer et al., (1994) conducted an experiment to determine the effectiveness of various
doses of propylene glycol in reducing plasma NEFA concentrations during restriction of


147
Table 5-1. Diet composition of far-off dry cow, transition dry cows and lactating
transition cows
Feed
Dry cows
far-off
Dry-cows
transition
Lactating
transition
Alafalfa hay
-
19.30
17.53
Coastal hay
-
4.76
-
Cottonseed whole
-
9.84
8.75
Com silage
44.90
24.06
28.50
Ryegrass silage
15.29
-
4.20
Com hominy
-
6.98
13.54
Citrus pulp
21.20
9.82
4.25
Soybean meal 48
17.20
3.44
5.49
Wet brewers grain
-
10.74
4.07
Lactating concentrate
-
-
7.03
Lactowhey 1
-
3.76
4.23
Metaxerol 2
-
2.79
1.96
Springer minerals
-
4.51
-
Dry cow minerals
1.41
-
-
Lactating inerals
-
-
0.45
1 Ammoniated whey (61.5% CP)
2 Energy supplement based on sodium propionate, propylene glycol, dried whey and
calcium carbonate


3-5. Reproductive responses for cows treated with or without monensin 109
4-1. Diet composition dry cow far-off, dry cow transition and lactating
transition cows 127
4-2. Nutrient content dry cow far-off, dry-cow transition
and lactating transition diets 128
4-3. Least Squares Means S.E.M. for rumen pH, acetate, propionate,
butyrate, valerate, total VFA, ammonia, D and L-lactate and
Acetate: Propionate by time in primiparous treated and control cows
at 10 d pp 129
4-4. Least Squares Means S.E.M. for rumen pH, acetate, propionate,
butyrate, valerate, total VFA, ammonia, D and L-lactate and
Acetate: Propionate by time in multiparous treated and control cows
at 10 d pp 130
4-5. Least Squares Means S.E.M. for serum 13-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling after
a.m. feeding in primiparous treated and control cows at 10 d pp 131
4-6. Least Squaresd Means S.E.M. for serum 13-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling after
a.m. feeding in multiparous treated and control cows at 10 d pp 132
4-7. Least Squares Means S.E.M. for serum 13-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling in
primiparous treated and control cows between dry-off and 21 d pp 133
4-8. Least Squares Means S.E.M. for serum 13-hydroxy butyrate,
non-esterified fatty acids and glucose by time of sampling in
multiparous treated and control cows between dry-off and 21 d pp 134
5-1. Diet composition dry cow far-off, dry cow transition and lactating
Transition cows 147
5-2. Nutrient content dry cow far-off, dry-cow transition and lactating
transition diets 148
5-3. Least Squares Means S.E.M. of ME305 and Real Milk Yield
(kg/lactation) by treatment and parity 149
6-1. Diet composition dry cow far-off, dry cow transition and lactating
Transition cows 163
6-2. Nutrient content dry cow far-off, dry-cow transition and lactating
VIII


48
concentration of plasma NEFA is maximized on day 1 after calving (Vazquez-Aon et al.,
1994). One explanation for this phenomenon is that the rate of hepatic acid esterification
exceeds the rate of TG disappearance via hydrolysis plus the export of newly synthesized
TG as very low density lipoprotein (Grummer, 1993; Hocquette and Bauchart, 1999).
Identification of the biochemical mechanism that limits efficient hepatic oxidation of fatty
acids remains elusive (Goff and Horst, 1997b). One of the theories was that microsomal
TG transfer protein (MPT), which is responsible for transfer of TG into the growing
VLDL particle, might be deficient or inactive in ruminant liver. However, Bremmer et al
(2000) found no correlation between MTP activity, mass, or mRNA with either liver TG
or plasma NEFA on d 2 postpartum. They concluded that MTP probably does not play a
role in the etiology of fatty liver that occurs in dairy cows at calving. Fatty liver can occur
very rapidly. Within 48 hours, hepatic TG levels can increase from less than 5% wet
weight to more than 25% under conditions of extreme adipose mobilization (Gerloff and
Herdt, 1999).
Ketosis cases that develop later during lactation, near the time of peak milk
production, may be of a different metabolic type. Gluconeogenic substrates are simply
insufficient to meet the demands of milk production. This results in high NEFA
concentrations, with a large portion being directed into ketogenesis, rather than
esterification. This kind of ketosis is not responsive to experimental glucagon therapy
(Herdt and Gerloff, 1999). Plasma glucose increased 11.5 and 9.0 mg/dl during week 1
and 2 after glucagon infusions. Nonesterified fatty acids and BHB were not affected (She
et al., 1999; Hippen et al., 1999a). However, in another experiment, Hippen et al. (1999b),
demonstrated that glucagon decreased the degree of fatty liver in early lactation and the
incidence of ketosis after alleviation of fatty liver. Membrane fatty acid composition in


University of Chile. Since that time, he was involved in several research projects and
teaching. In 1995, he visited the Swedish University of Agricultural Science to carry out
a training program in reproductive diseases of cattle. To these days that contact
continues. In 1996, Dr. Melendez made contact with a University of Floridas faculty
member. Initially, Dr. Melendez wanted to explore the possibility of doing a Master
Program. However, that UF faculty member, Dr. Carlos Risco, encouraged him to apply
to a residency program in Food Animal Production Medicine.
Circumstances or destiny, he started the residency program in June 1997 and his
master in June 1998. He obtained his Master of Science degree in 2000. He is now, with
his wonderful wife (Maria Ester) and his great sons (Diego and Ignacio) and daughter
(Elisa), finishing his PhD program. During his PhD program, Dr. Melendez obtained a
GPA of 4.0 and the Alec Courtelis Award in 2003, as one of the best international
graduate students within 6,000 foreign students. This prize also belongs to his adviser and
everybody who always supported and believed in Dr. Melendez.
193


NEFA (mEq/L) Glucose (mmol / L) BHB (mmol/L)
136
3
2.75 -
2.5 -
2.25 -
2
Monensin
p < 0.1
Control
Figure 4-2. Effect of monensin treatment on glucose, BHB, and NEFA concentration in
primiparous cows as a function of time after feeding at 10 d pp


176
Gilbert, R. O., and W. S. Schwark. 1992. Pharmacological considerations in the management
of peripartum conditions in the cow. Vet. Clin. North Am. Food Anim. Pract. 8:29-56.
Gillund, P., Reksen, O., Grdhn Y. T., and Karlberg, K. 2001. Body condition related to
ketosis and reproductive performance in Norwegian dairy cows. J. Dairy Sci. 84:1390-1396.
Goff, J. P. 1999. Treatment of calcium, phosphorus, and magnesium balance disorders. Vet.
Cl. North Amer. Food Anim. Pract. 15: 619-639.
Goff, J. P. 2000. Determining the mineral requirement of dairy cattle. Pages 106 132. In
Proceedings 11th Annual Florida Ruminant Nutrition Symposium, University of Florida.
Gainesville, FL.
Goff, J. P., R. L. Horst. 1993. Oral administration of calcium salts for treatment of
hypocalcemia in cattle. J. Dairy Sci. 76:101-108.
Goff, J. P., and R. L. Horst. 1997a. Effects of the addition of potassium or sodium but not
calcium, to prepartum rations on milk fever in dairy cows. J. Dairy Sci. 80: 176-186.
Goff, J.P., and R.L. Horst. 1997b. Physiological changes at parturition and their relationship to
metabolic disorders. J. Dairy Sci. 80:1260-1268.
Goff, J. P., and R. L. Horst. 1998a. Factors to concentrate on to prevent periparturient disease
in the dairy cow with special emphasis on milk fever. Proceedings 31th Conference of
American Association of Bovine Practitioners, Spokane, WA ,31:154-163.
Goff, J. P., and R. L. Horst. 1998b. Use of hydrochloric acid as a source of anions for
prevention of milk fever. J. Dairy Sci. 81:2874-2880.
Goff, J. P., R. L. Horst, P. W. Jardon, C. Borelli, and J. Wedam. 1996. Field trials of an oral
calcium propionate paste as an aid to prevent milk fever in periparturient dairy cows. J. Dairy
Sci. 79:378-383.
Goff, J. P., R. L. Horst, F. J. Mueller, J. K. Miller, G. A. Kiess, and H. H. Dowlen. 1991.
Addition of chloride to a prepartal diet high in cations increases 1,25-dihydroxyvitamin D
response to hypocalcemia preventing milk fever. J. Dairy Sci. 74:3863-3871.
Goff, J. P., M. E. Kehrli, and R. L. Horst. 1989. Periparturient hypocalcemia in cows:
Prevention using intramuscular parathyroid hormone. J. Dairy Sci. 72:1182-1187.
Grant, R. J., and J. L. Albright. 1995. Feeding behavior and management factors during the
transition period in dairy cattle. J. Anim. Sci. 73:2791-2803.
Green, B. L., B. W. McBride, D. Sandals, K. E. Leslie, R. Bagg, and P. Dick. 1999. The
impact of a monensin controlled-released capsule on subclinical ketosis in the transition dairy
cows. J. Dairy Sci. 82:333-342.


121
Rumen and Blood Sampling
At 10 days postpartum each experimental cow was subjected to a rumen and blood
sampling protocol. The first sample was obtained at 7 A.M. just before receiving the first meal
of the day. A second, third and fourth sample were obtained at 2, 4 and 6 h after feeding (2 h
intervals).
Each cow was placed in a special chute. A blood sample was taken as described
previously. Rumen samples were taken using a rumen tube sampling device (Jorgensen
Laboratories Inc., Loveland, CO). The distal end of the tube has a weighted metal device that
penetrates the rumen mat allowing sample collection from the ventral sac of the rumen. The
other end is connected to a manual vacuum pump that aspirates rumen fluid. A volume of
approximate 300 ml of rumen fluid was collected at each time. Rumen pH was measured
immediately after taking the sample with an electronic pH meter ((Micro pHep3 waterproof pH
tester, Hannah Instruments, Woonsocket, RI). Twenty ml of the ruminal fluid were placed in
plastic tubes and 5 ml of 20% phosphoric acid added and mixed with the sample to abruptly
cease rumen fermentation. Tubes were sealed and stored at 5 C until analysis.
Laboratory Analysis
Blood samples were centrifuged at 4000 rpm for 10 minutes. Plasma and serum were
separated and stored in plastic tubes and frozen at 20C until analysis was performed.
Serum non-esterified fatty acids (NEFA), P-hydroxy butyrate (BHB) and glucose were
measured. Non-esterified fatty acid concentration was determined by an enzymatic-
colorimetric method (Johnson and Peters, 1993) using a commercial kit (NEFA-C kit WAKO,
Saitama, Japan). Beta-hydroxybutyrate concentration was determined by an enzymatic-
colorimetric method (Williamson and Mellanby, 1974) using a commercial kit (Pointe


119
(Nagaraja et al., 1997; Stewart et al., 1997), it is hypothesized that in transition dairy cows fed
monensin and diets containing citrus pulp the ionophore still would affect the molar proportion
of VFA, NH3, lactate, rumen pH and blood metabolites in Holstein dairy cows.
In order to determine the effect of a monensin-controlled release capsule inserted
intraruminally at dry-off on the molar proportion of VFA, NH3, lactate, rumen pH and energy-
mineral related blood metabolites in Florida transition dairy cows fed citrus pulp-based diets
the following study was conducted.
Materials and Methods
Cows and Herd Management
The study was conducted on a commercial Holstein dairy farm with 3600 milking cows
located in north central Florida, with a milk rolling herd average of 10,700 kg. Most lactating
cows were housed in a dry-lot system and fed the same total mixed ration (TMR) three times a
day, except postpartum transition cows which received a diet higher in forage NDF. Cows
were dried-off between 50 to 70 days before expected parturition (BEP) and maintained in a
dry-lot (far-off cows) until 21 days BEP. They were fed a typical Florida dry cow diet (Table
4.1 and 4.2). Close-up dry cows (21 d BEP to calving) were housed in a dry-lot with adequate
feed-bunk space, water and shade. They received twice a day, a TMR containing citrus pulp
with a DCAD of -58.5 mEq/kg DM using the equation DCAD (mEq) = (Na + K) (Cl + S)
(Tables 4.1 and 4.2).
After calving, cows were moved from the processing pen to a postpartum lot and fed a
diet higher in forage NDF (Tables 4.1 and 4.2).


179
Herdt, T. H. 2002. Gastrointestinal physiology and metabolism. Postabsorptive nutrient
utilization. Pages 303-322 in Textbook of Veterinary Physiology. Third Edition. J.
Cunningham. W.B. Saunders Company
Herdt T. H., R. S. Emery. 1992. Therapy diseases of ruminant intermediary metabolism. Vet.
Clin. North Am. Food Anim. Pract. 8: 91-106
Herdt, T.H. and B.J. Gerloff. 1999. Ketosis. Pages 226-230 in Current Veterinary Therapy 4.
Food Animal Practice. J. Howard and R. Smith. W.B. Saunders Company.
Heuer, C., Y. H. Schukken, and P. Dobbelaar.1999. Postpartum body condition score and
results from the first test day milk as predictors of disease, fertility, yield, and culling in
commercial dairy herds. J. Dairy Sci. 82:295-304.
Heuer, C., Y. H. Schukken, L.J. Jonker, J.I.D. Wilkinson and J.P.T.M. Noordhuizen 2001.
Effect of monensin on blood ketone bodies, incidence and recurrence of disease and fertility
in dairy cows. J. Dairy Sci. 84:1085-1097.
Heuer, C., A. Wangler, Y.H. Schukken, and J.P.T.M. Noordhuizen. 2001. Variability of
acetone in milk in a large low-production dairy herd: a longitudinal case study. The Vet. J.
161:314-321.
Higgins, J. J., W. K. Sanchez, M. A. Guy, and M. L. Anderson. 1996. An oral gel of calcium
propionate plus propylene glycol is effective in elevating calcium and glucose levels in
periparturient dairy cows. J. Dairy Sci. 79(Suppl. 1): 130.
Hippen, A. R. 2000. Glucagon as a potential therapy for ketosis and fatty liver. Vet. Clin.
North Am. Food Anim. Pract. 16:267-282.
Hippen, A. R., P. She, J. W. Young, D. C. Beitz G. L. Lindberg, L. F. Richardson, and R. W.
Tucker. 1999a. Metabolic responses of dairy cows and heifers to various intravenous dosages
of glucagon. J. Dairy Sci. 82:1128-1138.
Hippen, A. R., P. She, J. W. Young, D. C. Beitz G. L. Lindberg, L. F. Richardson, and R. W.
Tucker. 1999b. Alleviation of fatty liver in dairy cows with 14-day intravenous infusions of
glucagon. J. Dairy Sci., 82:1139-1152.
Hoblet, K. H., G. D. Schnitkey, D. Arbaugh, J. S. Hogan, K. L. Smith, P. S. Schoenberger, D.
A. Todhunter, W. D. Hueston, D. E. Pritchard, G. L. Bowman, L. E. Heider, B. L. Brocket,
and H. R. Conrad. 1991. Cost associated with selected preventive practices and with episodes
of clinical mastitis in nine herds with low somatic cell counts. J. Amer. Vet. Med. Assoc.
199:190-196.
Hobson, P.N. 1997. The functioning rumen. Pages 5-9 in The rumen microbial ecosystem.
Second edition. P.N. Hobson and C.S. Stewart. Blackie Academic & Professional.


141
Monensin is not approved for use in lactating dairy cattle in the United States, however
it is permitted in Canada and Australia. The use of a monensin-controlled release capsule has
been demonstrated to be useful in the prevention of calving-related disorders in transition dairy
cows (Stephenson et al., 1997; Duffield et al., 1998b, Duffield et al., 1999b; Green et al., 1999;
Mutsvangwa et al., 2002).
Since pectin-fermenting bacteria are gram-negative monensin-resistant bacteria
(Nagaraja et al., 1997; Stewart et al., 1997), it is hypothesized that in dairy cows fed monensin
and diets containing citrus pulp pectin digestion should not modified extensively. As a result,
monensin should still positively affect energy related blood metabolites in transition dairy cows.
Therefore, the objective of this study was to determine the effect of a monensin-controlled release
capsule inserted at dry-off on milk yield, BCS at calving and concentrations of NEFA, BHB and
glucose during the transition period, in Florida dairy cows fed diets containing citrus pulp.
Materials and Methods
Cows and Herd Management
The study was conducted on a commercial dairy farm with 3600 milking cows located in
north central Florida with a milk rolling herd average of 10,700 kg. Most lactating cows were
housed in a dry-lot system and fed the same TMR three times a day, except postpartum transition
cows which received a diet higher in forage NDF. Cows were dried-off between 50 to 70 days
before expected parturition (BEP) and maintained in a dry-lot (far-off cows) until 21 days BEP.
They were fed a typical Florida dry cow diet (Table 5.1 and 5.2) and moved 21 days BEP to a
different pen to initiate their prepartum transition period. They were housed in a dry-lot with
adequate feed-bunk space and shade. Twice a day they received a diet containing citrus pulp with a
DCAD of -51.7 mEq/kg DM using the equation DCAD (mEq)= (Na + K) ( Cl + S) (Tables 5.1
and 5.2).


66
by methanogenic bacteria and increased heat increment and the apparent digestibility of
metabolizable energy (Wedegaertner and Johnson, 1983; Oscar and Spears, 1990).
Monensin affects nitrogen metabolism by decreasing ammonia production in
cattle fed a forage -based diet. This might be a result of reduced proteolysis, degradation
of peptides and deamination of amino acids in the rumen (Yang and Russell, 1993;
Nagaraja et al., 1997). Under monensin, the specific activity of ammonia production by
mixed ruminal bacteria is decreased by more than 30% and this decrease corresponds to
about a 10-fold decrease in the numbers of bacteria that ferment peptides and amino acids
as an energy source for growth (Yang and Russell, 1993). Ionophores have also been
demonstrated to decrease ruminal urease activity (Starnes et al., 1984).
Monensin prevents lactic acid build-up in the rumen by its selectivity towards
gram-positive bacteria. The major lactic acid-producing bacteria (S. bovis and
Lactobacillus ssp.) are inhibited while ruminal lactic acid-fermenting bacteria (gram
negative) are unaffected (Nagaraja et al., 1997). This effect was not repeated in a more
recent study that used a monensin controlled-release capsule or a premix in dairy cattle
(Mutsvangwa et al., 2002).
Monensin also decreases the incidence of frothy bloat as a result of reduction in
microbial slime and gas production (Lowe et al., 1991; Nagaraja et al., 1997). Monensin
reduces or modulates feed intake in both grain-fed and forage-fed cattle, probably due to
low palatability or decrease in rumen turnover rate of liquids and solids, and consequently
increase of ruminal fill (Stock et al., 1995; Nagaraja et al., 1997).


Body Condition Score (1 -5)
150
3.5
3.25 -
3
2.75
P < 0.05
*
M Control
Primi parous
- A- Monensin
Primi parous
K Control
Multiparous
* Monensin
Multiparous
I
Dry-Off
Calving
Figure 5-1. Body Condition Score by treatment and by parity at assignment
and at calving


151
20 1 1 1 1 1 1 1 1 1 1
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10
DHIA Tesd Day
Figure 5.2 Test day milk production by treatment in primiparous cows


83
Nearly all of the dietary K is absorbed. Intracellular uptake of K following a meal
helps buffer blood K concentration. The factorial approach as described for Ca, P and Mg
does not work for Na, K and Cl (Goff, 2000)
Rumen microbes need 0.2-0.22% S to operate efficiently. Excessive dietary S can
interfere with absorption of Cu and Se and may become toxic (Goff, 2000).
Cobalt is a component of vitamin B12. Microbes in rumen are the only natural
source of vitamin B12. Rumen microbes need 0.11% Co to perform efficiently (Goff,
2000).
Between 1 and 5% of dietary Cu will be absorbed by adult cattle. A diet high in Zn
(>1000 mg/kg), S and Mo can block Cu absorption. The Cu:Mo ratio should be 2:1 (Goff,
2000).
Selenium is part of glutathione peroxidase and in conjunction with vitamin E act
as antioxidant compounds. Selenium is also critical to thyroid hormone metabolism.
Selenium can cause deficiency or toxicity. Diets containing 0.1 ppm of Se are
recommended but field studies suggest this is not enough. Legally Se can be added up to
0.3 ppm (Goff, 2000).
Depending on the DMI the dietary requirement of iodine should be about 0.25-0.5
mg/kg DM (Goff, 2000).
Iron excess (250-500 mg Fe/kg DM) interfere with Cu and Zn absorption. NRC
(1989) recommends dietary Fe not to exceed 1000 mg/kg DM. This may be too high
(Goff, 2000).
Chromium is essential for normal glucose metabolism. The amount of Cr required
in the diet for optimal performance is unclear. More research is required (Goff, 2000).


81
not benefit from a diet with more than 12.7% CP. In other studies, feeding higher protein
during the transition period reduced feed intake or milk yield postpartum (Crawley and
Kilmer, 1995; Donkin et al., 1998).
day relative to calving
Figure 2-3. Estimated protein density required in diets of transition cows to meet
requirements for maintenance and gestation according to NRC,
(2001)


CHAPTER 1
INTRODUCTION
The transition period, defined as the last 3 weeks before parturition to 3 weeks after
parturition is the most stressful and challenging stage of the lactation cycle (Grummer,
1995; Drackley, 1999). During the last 10 years, research on this topic have been copious,
and for first time the National Reseach Council (NRC, 2001) has included a chapter
covering the most important and unique aspects of transition dairy cows. Since most of the
metabolic disorders of dairy cows occur during this time (Goff and Horst, 1997b;
Drackley, 1999), strategies to prevent calving-related disorders have been focused on the
nutritional and feeding management aspects of the prepartum transition dairy cow (Goff
and Horst, 1997b; Gerloff, 2000; Oetzel, 2000).
The use of feed additives has become a common practice in dairy cattle. They
improve palatability, enhance rumen fermentation, optimize rumen digestion of the fiber,
reduce mobilization of adipose tissue, and maintain calcium and phosphorus homeostasis
during the first few days of lactation (Santos, 1999). However, fed additives do not replace
the optimum levels of nutrients, but complement the effect of a good dietary management.
Unbalanced diets or poor management will not be corrected by the use of additives
(Gerloff, 2000).
Sodium monensin is an additive ionophore antibiotic that selectively modifies the
ruminal flora and improves the digestive efficiency in cattle, increasing productive
parameters and decreasing health problems. Monensin increases ruminal propionate
1


84
The dietary coefficient of absorption for Zn is estimated to be 0.15. The major
dietary factors that can modify the efficiency of absorption of dietary Zn are interactions
of Zn with other metal ions (Cu, Fe) and the presence of organic chelating agents in the
diet (Goff, 2000).
The proportion of Mn absorbed from the diet generally is between 0.5 and 1%.
High dietary Ca, K or P increase Mn excretion in feces (Goff, 2000).
Mahanna (1997) recommends 0.3 ppm of Se dry matter basis. Potassium should
be not over 1.0% in the total ration for dry cows, but K Na and Mg should be 1.5, 0.5
and 0.35% of DM respectively in milking cows during heat stress. Nitrogen to sulfur ratio
should be 11 to 13:1 in the total ration to meet rumen bacterial needs. Goff and Horst
(1998a) recommend that dietary magnesium should be set at 0.4% (higher than NRC,
1989). Magnesium sulfate or magnesium chloride is recommended because they could be
an effective source of anions and magnesium.
Furthermore, the diet should supply between 35 to 50 g of phosphorus daily
(-0.4%). More than 80 g of P/day will inhibit renal synthesis of 1,25-dihydroxyvitamin D
which can induce MF. Dietary S should not exceed 0.4%. Adding more sulfate is a poor
choice because it is fairly ineffective acidifying agent. Dietary Cl can nearly always be
raised to 0.5% with little effect on dry matter intake. High dietary Ca concentrations (1.0-
1.2%) are desirable with anionic salts. Good results have been achieved by feeding as high
as 180 g Ca/day In one study, Joyce et al. (1997) determined that pre-calving cows fed
diets based on alfalfa (typically high in Ca) with a DCAD of 7 had the lowest urine pH
prepartum and the highest concentrations of ionized Ca in blood. Also, this group
experienced the lowest incidence of calving related disorders. Results indicated that alfalfa
when supplemented with anionic salts is a viable forage for prepartum dairy cows.


50
subclinical ketosis (Duffield, 2000). A threshold of 500 pmol/L (5.0 mg/dL) of blood
acetoacetate in cows with clinical ketosis has been reported (Baird, 1982). This threshold
would approximate 3.9 mmol/L (40 mg/dL) of BHB.
Potential therapeutic approaches to fatty liver include (1) limiting the fatty acid
supply by limiting adipose mobilization, (2) limiting hepatic fractional extraction (uptake)
of fatty acids, (3) increasing oxidative metabolism of fatty acids, or (4) increasing the rate
of triglyceride secretion from the liver (Herdt and Emery, 1992). Treatment of ketosis is
based on agents that reestablish a normal appetite and restore normal blood concentrations
of glucose and ketone bodies (Herdt and Gerloff, 1999).
Administration of 500 mL of 50% glucose or dextrose solution, intravenously, is a
common therapy for bovine ketosis (Herdt and Emery, 1992). Other therapeutic
alternatives are glucose precursors. They can be administered orally to ruminants. These
include glycerol, propylene glycol and propionate. Glucocorticoid therapy is also
effective. Ketotic cows treated with glucocorticoids are less subject to relapses than are
those treated with IV glucose alone, although relapses can still occur (Herdt and Gerloff,
1999). Dexamethasone and 9-Flurprednisolone acetate at a dose of 1.33 mg/45 kg body
weight and Flumethasone at a dose of 0.33 mg/45 kg of body weight are recommended
(Herdt and Emery, 1992). Insulin, in conjuction with glucocorticoids, may be a more
effective therapy. Insulin is a powerful antiketogenic agent and also suppresses NEFA
mobilization. A long acting form of insulin should be used at a dose of 200 to 300 IU per
animal, repeated as necessary at 24 to 48 hours intervals. Glucocorticoids or other agents
have to be provided to counteract the hypoglycemic effects of insulin (Herdt and Gerloff,
1999).


28
Uterine infections can reduce the reproductive efficiency of dairy cows thereby
increasing herd health cost (Lewis, 1997). Metritis increased days from calving to first
estrus by 6.9 days, 7.3 days calving to first service interval, 15.4 days first to last service
interval, 0.3 services per conception and 18 days calving to conception interval (Barlett et
al., 1986a). Uterine infections alter uterine involution and reduce ovarian follicular
development during the early postpartum period, which may prolong the interval from
calving to estrus and AI, but the mechanism and the repeatability of the effect of the
condition on ovarian follicles is unclear (Del Vecchio et al., 1994; Lewis, 1997).
Uterine infections are a highly complex process. The exact causes of uterine
infections are unknown, but several predisposing factors have been associated with the
disease (Lewis, 1997). Cows with dystocia, RFM, twins or still-births, overconditioning
and various metabolic and digestive disorders increase the risk of metritis (Curtis et al.,
1985; Erb et al., 1985; Olson et al., 1986; Correa et al., 1993; Lewis, 1997; Youngquist
and Shore; 1997; Bruun et al., 2002; Melendez et al., 2003b). Other risk factor that might
be associated with the pathogenesis of metritis is the immunosupression occurring around
parturition. T-cell sub-populations have been demonstrated to decline at calving and they
do not return to pre-calving levels until two weeks after parturition (Kimura et al., 1999).
Treatment of REM has been based on different protocols. Removal by gentle
traction has long been the conventional method, but the procedure may be followed by
severe uterine infections and impaired fertility (Bolinder et al., 1988). A single dose of
oxytocin does not reduce the presence of RFM (Miller and Lodge, 1984). Exogenous
estrogen is of questionable therapeutic value because plasma concentrations of estrogen
are elevated in cows when the placenta is retained (Pimental et al., 1987). Analogs of
Prostaglandin F2 alpha are recommended as well, but Fenprostalene does not change


I certify that I have read this study and that in my opinion it conforms to
acceptable standards of scholarly presentation and is fully adequate, in scope and
quality, as a dissertation for the degree of Doctor of Philosophy.
Arthur G. Donovan
Associate Professor of College
of Veterinary Medicine
I certify that I have read this study and that in my opinion it conforms to
acceptable standards of scholarly presentation and is fully adequate, in scope and
quality, as a dissertation for the degree of Doctor of Philosophy. /) 7 //I
Louis A. Archbald
Professor of College
of Veterinary Medicine
I certify that I have read this study and that in my opinion it conforms to
acceptable standards of scholarly presentation and is fully adequate, in scope and
quality, as a dissertation for the degree of Doctor of Philosophy.
^
Jorge Hernandez
Associate Professor of College
of Veterinary Medicine
I certify that I have read this study and that in my opinion it conforms to
acceptable standards of scholarly presentation and is fully adequate, in scope and
quality, as a dissertation for the degree of Doctor of Philosophy.
Carlos A. Risco
Professor of College
of Veterinary Medicine
I certify that I have read this study and that in my opinion it conforms to
acceptable standards of scholarly presentation and is fully adequate, in scope and
quality, as a dissertation for the degree of Doctor of Philosophy.
Ramon Littell
Professor of Statistics, IFAS


NE1, Mcal/kg
79
day relative to calving
Figure 2-2. Estimated energy density required in diets of transition cows to meet
requirements for maintenance and gestation according to NRC,
(2001)


181
Kaneene, J. B., R. A. Miller, T. H. Herdt, and J. C. Gardiner. 1997. The association of serum
nonesterified fatty acids and cholesterol, management and feeding practices with peripartum
disease in dairy cows. Prev. Vet. Med. 31:59-72.
Kelton, D. F., K. D. Lissemore, and R. E. Martin. 1998. Recommendations for recording and
calculating the incidence of selected clinical diseases of dairy cattle. J. Dairy Sci. 81:2502-
2509.
Kimura, K., J. P. Goff, M. E. Kehrli, and J. A. Harp. 1999. Phenotype analysis of peripheral
blood mononuclear cells in peripartum dairy cows. J. Dairy Sci. 82:315-319.
Kimura, K., J. P. Goff, M. E. Kehrli, and T. A. Reinhardt. 2002. Decreased neutrophil
function as a cause of retained placenta in dairy cattle. J. Dairy Sci. 85:544-550.
Laranja da Fonseca, L. F., C. S. Lucci, P. H. M. Rodriguez, M. V. Santos, and A. P. Lima.
1998. Supplementation of propylene glycol to dairy cows in periparturient period: Effects on
plasma concentration of BHBA, NEFA, and glucose. J. Dairy Sci. 81(Suppl. 1):320.
Lean, I. J., M. Curtis, R. Dyson, and B. Lowe. 1994. Effects of sodium monensin on
reproductive performance of dairy cattle. I. Effects on conception rates, calving to conception
intervals, calving to heat and milk production in dairy cows. Aust. Vet. J. 71:273-277.
Leiva, E., M.B. Hall, and H.H. Van Horn. 2000. Performance of dairy cattle fed citrus pulp or
com products as sources of neutral detergent-soluble carbohydrates. J. Dairy Sci. 83:2866-
2875.
Lewis, G.S. 1997. Uterine health and disorders. J. Dairy Sci. 80:984-994.
Littell, R. C., P. R. Henry, and C. B. Ammerman. 1998. Statistical analysis of repeated
measures data using SAS procedures. J. Anim. Sci. 76:1216-1231.
Lowe, L.B., G.J. Ball, B.R. Carruthrers, R.C. Dobos, G.A. Lynch, P.J. Moate, P.R. Poole, and
S.C. Valentine. 1991. Monensin controlled release intra-ruminal capsule for the control of
bloat in pastured dairy cattle. Aust. Vet. J. 68:17-20.
Ludvigsen, C.W., J. R. Thum, G.L. Pierpont, J.H. Eckfeldt. 1983. Kinetic assay for D (-)-
lactate, with use of a centrifugal analyzer. Clin. Chem. 29:1823-1825.
Mallard, B. A., J. C. Dekkers, M. J. Ireland, K. E. Leslie, S. Sharif, C. Lacey Vankampen, L.
Wagter, and B. N. Wilkie. 1998. Alteration in immune responsiveness during the peripartum
period and its ramification on dairy cow and calf health. J. Dairy Sci. 81:585-595.
Markusfeld, O. 1986. The association of displaced abomasum with various periparturient
factors in dairy cows. A retrospective study. Prev. Vet. Med. 4:172-183.


10
Table 2-2. Effect of hormones on carbohydrates and lipids intermediary metabolism on
dairy cattle.
Hormone
Effect on carbohydrates
Effect on lipids
Insulin
t glucose transport into cells
i gluconeogenesis
t glycogen synthesis
i glycogenolysis
t glycolisis
i lipolysis
t lipogenesis
Glucagon
t gluconeogenesis
t glycogenolysis
t glucose export
i glycolisis
i glycogen synthesis
t lipolysis
T ketogenesis ?
Catecholamines
t glycogenolysis
t gluconeogenesis
t glucagons secretion
i insulin secretion
f lipolysis
Growth
Hormone
t blood glucose
i lipogenesis
I NEFA mobilization
Cortisol
t gluconeogenesis from proteins
f lipolysis
Adapted from:
Herdt, T. H. 2000. Ruminant adaptation to negative energy balance. Influences on the
etiology of ketosis and fatty liver. Vet. Clin. North Am. Food Anim. Pract. 16:215-230.
Nelson, D. L. and M. M. Cox. 2000. Oxidation of fatty acids. Pages 598-622. In
Lehninger Principles of biochemistry. Third edition. Worth Publishers, New York, NY
10010.
Drackley, J. K., T. R. Overton, and G. N. Douglas. 2001. Adaptations of glucose and long-
chain fatty acid metabolism in liver of dairy cows during the periparturient period. J.
Dairy Sci. 84(E. Suppl.):E100-E112.
Herdt, T. H. 2002. Gastrointestinal physiology and metabolism. Postabsorptive nutrient
utilization. Pages 303-322 in Textbook of Veterinary Physiology. Third Edition. J.
Cunningham. W.B. Saunders Company


21
Parathyroid hormone administration increases plasma concentration of 1,25 dihydroxy
vitamin D and hydroxyproline prior to parturition, suggesting that both intestinal Ca
absorption and bone Ca resorption are increased by administration of the hormone (Goff et
al., 1989).
Dietary cation-anion difference has been defined as the difference in
milliequivalents of cations and anions per kilogram of dry matter and has a direct impact
on blood acid-base metabolism (Block, 1994). Important dietary cations are sodium (Na),
potassium (K), calcium (Ca) and magnesium (Mg); important dietary anions are chloride
(Cl), sulfur (S) and phosphorus (P). Several methods of calculating DCAD have been
utilized, including the following equations:
DCAD (mEq)= (Na + K) ( Cl + S) (3-1)
DCAD (mEq)= (Na + K) (Cl) (3-2)
DCAD (mEq)= (Na + K + 0.15 Ca + 0.15 Mg) ( Cl + 0.2 S + 0.3 P) (3-3)
DCAD (mEq)= (Na + K + 0.15 Ca + 0.15 Mg) ( Cl + 0.25 S + 0.5 P) (3-4)
Equations 3-1 and 3-2 are used more commonly, but it must be kept in mind that
Ca, Mg, and P absorbed from the diet will also influence blood pH. Equations 3-3 and 3-4
take into account new data on the bioavailability of all the potential strong ions.
Theoretically it should be the most accurate, but it is has not been widely applied (Goff
and Horst, 1998a; Oetzel and Goff, 1999).
In lactating dairy cows, increasing dietary cation-anion difference increased DMI
and milk production in early and midlactation. These effects were not observed in late
lactation. However, increased DCAD affected acid-base parameters in urine at all stages
of lactation (Delaquis and Block, 1995).


148
Table 5-2. Nutrient content of far-off dry cow, transition dry cows and lactation
transition diets
Nutrient
Dry cow
far-off
Dry cow
transition
Lactating
transition
CP(%DM) 1
15.52
17.91
18.60
UndegP(%CP)2
26.38
35.31
30.34
DegP(%CP) 2
73.62
64.69
69.66
SolP(%CP) 2
-
36.72
39.77
NEL(Mcal/kg)3
0.84
1.69
1.69
ADF(%DM) 1
24.54
25.13
23.66
NDF(%DM) 1
36.04
36.20
34.63
NFC(%DM) 2
32.94
31.55
34.13
Starch(%DM) 2
11.05
14.96
14.90
Lipid (%DM) 1
2.85
6.20
2.36
Ca(%DM) 1
0.74
1.27
1.10
P(%DM) 1
0.30
0.35
0.46
Mg(%DM) 1
0.32
0.36
0.36
K(%DM) 1
1.28
1.10
1.46
Na(%DM) 1
0.09
0.09
0.58
C1(%DM) 1
0.18
0.43
0.48
S(%DM) 1
0.24
0.40
0.22
Forage in diet (%DM)
60.21
48.10
50.23
Cation-Anion (meq/kg DM) 4
166.0
-52.0
352.0
1 Laboratory nutritional analysis
2 Values from feed composition tables
1 From formulas after laboratory analysis
4 From formula (Na+ + K+) (CF + S')


NEFA (mEq/L) Glucose (mmol / L) BHB (mmol/L)
153
Figure 5-4. Glucose, beta-hydroxy butyrate (BHB), and non-esterified fatty acids
(NEFA) concentrations over time in monensin treated and control
primiparous cows


100
In this study, correcting for other variables, treatment was associated with a lower
incidence of metritis. Dystocia and parity were also associated with metritis (Table 3.4).
An explanation for the positive effect of treatment on metritis is challenging. Perhaps
treated cows had a better metabolic status postpartum with less fat mobilization and less
BHB production than control cows. In addition, treated cows had a higher BCS at
calving (not obese) than control cows, which might have influenced positively cow
performance in the early postpartum period. Indeed, cows in good BCS at calving (3.25-
3.5) that maintained their condition through optimal feed intake in the early postpartum
period best adjusted metabolically to increased energy requirements (Busato et al.,
2002). Finally, monensin has been demonstrated to reduce the rate of intramammary
infections in lactating dairy cows suggesting that monensin through its effect on
metabolism can decrease immunosuppression in early lactation (Heuer et ah, 2001).
The incidence of clinical ketosis reported in this study was in the lower limit of
the incidence range of 2 to 22% reported by other authors (Grohn et ah, 1998; Kelton et
ah, 1998; .Grohn et ah, 1999; Gstergard and Grohn, 1999; Al-Rawashdeh, 1999).
However, the main parameter used to diagnose ketosis in this trial was a decrease in
milk yield and detectable concentration of ketone bodies in urine using a colorimetric
field test based on nitropruside (Ketostix). This might be an underestimation of the real
incidence of the disease because Ketostix presents a low sensitivity (Geishauser et ah,
1998). There was no effect of treatment detected although monensin has been shown to
be an antiketogenic product in dairy cattle (Duffield, 1997; 1998a; 1998b; 1999b; 2003).
However, most of the Canadian studies have been focused on the effects of the bolus on
subclinical ketosis rather than clinical ketosis and one of these studies found no
differences on the incidence of clinical ketosis between treated and control cows


TABLE OF CONTENTS
page
ACKNOWLEDGMENTS iii
ABSTRACT xii
CHAPTER
1 INTRODUCTION 1
2 LITERATURE REVIEW 4
The Transition Period 4
Physiological and Metabolic Changes during the Transition
Period 5
Dry Matter Intake 5
Glucose and Lipid Metabolism 6
Calving-Related Disorders 8
Milk Fever, Parturient Paresis, Hypocalcemia 15
Retained Fetal Membranes-Metritis Complex 23
Abomasal Disorders 30
Mastitis 37
Lameness 38
Culling 40
Ketosis-Fatty Liver Complex 42
Monensin 59
Rumen Microbial Ecosystem 60
Volatile Fatty Acids 65
Monensin and Rumen Fermentation 65
Citrus Pulp and Pectin Fermentation 68
Monensin, Calving-Related Disorders, Reproductive
and Productive Responses 69
Transition Cow Feed Management 77
Energy 78
Protein 80
Minerals and Vitamins 82
Dry Cow Feed Management and Body Condition Score 85
Fresh Cow Feed Management and Body Condition Score 86
v


86
separate close-up group for pregnant heifers might be a beneficial management strategy in
farms if adequate facilities are available (Grant and Albright, 1995). Diet formulation
should be based on 10 to 11 kg of DM intake. If cows are overeating it is not a big
problem because this is a short period of time. Cows should be in a positive energy status
and not losing weight. Feed should always be available in the bunk (24 h a day) in a form
of TMR to ensure adequate control of nutrient composition and consumption. Fermentable
grains (coarsely ground dry com, hominy, or high moisture com) to enhance development
of the ruminal epithelia should be fed (Beede, 1997). Cows should calve with a BCS of
3.5-3.75 and heifers 3.25-3.5 to minimize obstetrical trauma (Beede, 1997; Studer, 1998;
Grummer, 1999).
Fresh Cow Feed Management and Body Condition Score
The primary goal for early fresh cows is to maximize carbohydrate, protein and
nutrient intake and provide adequate fiber to meet requirements for increasing milk
production (Beede, 1997).
Mahanna (1997) recommends that cows should reach maximun daily DMI no later
than 10 weeks postpartum. Cows milked three times a day should eat about 5 to 6% more
DM per day. For every 1 kg of expected milk production cows should eat at least 0.45 kg
of DM. Eating less than this causes excessive body condition loss. Forage DM intake
should be near 2% of the cows BW. Acid detergent fiber should be at least 19-21% and
NDF at least 28-30% or 0.9% of BW. Particle size should be long enough to stimulate 30
minutes of cud chewing time per kg of DM. Total mixed ration DM content should be
between 50 to 75%. Cows should reduce their DMI about 3.3% for every 1C over 24C.
Heat stress starts when the temperature exceeds 26.7 C and relative humidity exceeds


REFERENCE LIST
Abe, N., I. J. Lean, A. Rabiee, J. Porter, C. Graham. 1994. Effects of sodium monensin on
reproductive performance of dairy cattle. II Effects on metabolites in plasma, resumption of
ovarian cyclicity and oestrus in lactating cows. Aust. Vet. J. 71:277-282.
Agricultural Research Council. 1980. The nutrient requirements of ruminant livestock.
Slough, U.K.: Commonwealth Agricultural Bureaux
Allen, M. S. 2000. Effects of diet on short-term regulation of feed intake by lactating dairy
cattle. J. Dairy Sci. 83:1598-1624.
Al-Rawashded, O.F. 1999. Prevalence of ketonemia and associations with herd size, lactation
stage, parity and postparturient diseases in Jordanian dairy cattle. Prev. Vet. Med. 40:117-125.
Anderson, L. 1988. Subclinical ketosis in dairy cows. Vet. Clin. North Am. Food Anim. Pract.
4:233-251.
Andersson, L., A. H. Gustafsson, and U. Emanuelson.1991. Effect of hyperketonaemia and
feeding on fertility in dairy cows. Theriogenology 36:521-536.
Arthington, J.D., W.E. Kunkle, and A.M. Martin. 2002. Citrus pulp for cattle. Vet. Clin. North
Am. Food Anim. Pract. 18:317-326.
Baile, C.A., C.L. McLaughlin, W.V. Chalupa, D.L. Snyder, L.C. Pendlum and E.L. Potter.
1982. Effects of monensin fed to replacement dairy heifers during the growing and gestation
period upon growth, reproduction, and subsequent lactation. J. Dairy Sci. 65:1941-1944.
Baird, D.G. 1982. Primary ketosis in the high producing dairy cow: Clinical and subclinical
disorders, treatment, prevention and outlook. J. Dairy Sci. 65:1-10.
Barlett, P. C., J. H. Kirk, M. A. Wilke, J. B. Kaneene, and E. C. Mather. 1986a. Metritis
complex in Michigan Holstein-Friesan cattle: incidence, descriptive epidemiology and
estimated economic impact. Prev. Vet. Med. 4:235-248.
Barlett, P. C., G. Y. Miller, C. R. Anderson, and J. H. Kirk. 1990. Milk production and
somatic cell count in Michigan dairy herds. J. Dairy Sci. 73:2794-2800.
Bauchemin, K. A. 1991. Indigestion and mastication of feed by dairy cattle. Vet. Clin. North
Am. Food Anim. Pract. 7:439-464.
169


122
Scientific, Inc. BHA Set. Lincoln Park, MI, USA). Glucose concentration was determined
using a kit based on Trinder reaction (Sigma, St. Louis, MO) (Bergmeyer and Bemt, 1974).
Rumen samples were analyzed for concentrations of acetic, propionic, butyric and, L
and D-lactic acids and NH3. Volatile fatty acid concentrations were determined through gas
chromatography separation technique (Supelco Inc., Bellefonte, PA 16823). D and L-lactate
concentration were determined enzymatically by the D and L-lactate dehydrogenase reaction
(Olsen, 1962; Ludvigsen et al., 1983) using a micro-centrifugal analyzer. Ammonia levels were
measured by the glutamate dehydrogenase reaction (Bergmeyer and Beutler, 1985) using a
micro-centrifugal analyzer.
Statistical Analysis
The null hypothesis was that there is no difference in the concentration of rumen and
blood metabolites between treatment groups. Data for rumen and blood metabolites were
analyzed by ANOVA, constructing mixed models for repeated measures.
Variables were considered significant at P < 0.05. Statistical analysis was conducted
using SAS 8.2 (SAS, 2001).
Mixed models for repeated measures were defined as:
y¡jk= p + Tj + Timek + Pari +(Time*T)ki + (Par *Time*T)ki| + e¡jki
Where:
yijicP Blood, rumen metabolites, BCS
Ti= fixed effect of treatment
Cow (Tj)j = random effect of cow nested in treatment
Time k = fixed effect of time
Par i = fixed effect of parity
(Time *T) ¡k = interaction time and treatment
(Par *Time*T) k¡i = interaction time, parity and treatment
eijki = random error term


164
transition diets
6-3. Receiver-operating characteristic (ROC) analysis for milk BHB
at three different levels of serum BHB (gold standard) 165
6-4. Sensitivity, specificity, positive predictive value, negative
predictive value of milk BHB test based on a serum BHB
cut-off value of 1.0 mmol/L 166
6-5. Summary of logistic regression modeling 167
IX


109
Table 3.5. Reproductive responses for cows treated with or without monensin
Variable
Control
(n=290)
Monensin
(n=290)
Conception rate at first service (%)
.
42.4
5lT
Overall pregnancy rate (%)
53.9
58.1
Services per conception
2.68 0.35
2.36 0.35
Calving to conception interval (days)
180.2 12.2
170.4 12.2
P = 0.12


NEFA (mEq/L) Glucose (mmol / L) BHB (mmol/L)
139
l -i
0.9 -
0.8 -
0.7 -
0.6 -
0.5 -
0.4 -
0.3 -
0.2 -
0.1 -
0 -
1.5
1.3
1.1
0.9
0.7
0.5
0.3
0.1
Dry-off Close-up Calving 7 d pp 14 d pp 21dpp
Control
T 1 1
Figure 4-5 Effect of monensin treatment on glucose, BHB, and NEFA concentration
in multiparous cows as a function of time


59
Monensin
Ionophores are compounds that collapse the ion gradients across cellular
membranes. Those that are specific for microorganisms can serve as antibiotics. These
compounds shuttle ions across the membranes and kill microbial cells by disrupting
secondary transport processes and energy-conserving reactions (Nelson and Cox, 2000).
Monensin is one of these antibiotics that act as a sodium carrying ionophore. It is a cyclic
peptide that surrounds Na+ and neutralizes its positive charge. The peptide carries Na+
across membranes down its concentration gradient and deflates that gradient (Nelson and
Cox, 2000). The ionophore-cation complex attaches to bacteria and becomes solubilized in
the lipid bilayer of the cell membrane. Once solubilized in the cell membrane, the
complex cation is exchanged for a proton. This culminates in a greater intracellular Na+
concentration, reduced intracellular K+ concentration and lower intracellular pH. Finally,
bacteria are forced to utilize cellular transport systems to dissipate the intracellular H+ and
Na+. This reduces energy reserves, resulting in lowered capability for cell division and
protein synthesis. Cell death comes from acidification of the cytoplasm (McGuffey et al.,
2001).
Monensin was discovered in an in vitro rumen batch fermentation screening
program (Richardson et al., 1976). Monensin enhances propionate production in a high-
roughage and high-grain diets by 49% and 76%, respectively (Van Maanen et al., 1978).
Prange et al. (1978) also demonstrated that monensin enhanced rumen propionic acid
production by 44% in Holstein steers fed a 30% concentrate diet. These studies confirmed
that the early observations of monensin-mediated in vitro rumen propionate production
and in vivo enhancements of rumen molar percentage of propionate are in fact indicative
of elevated in vivo propionic acid production rates. Monensin works by inhibiting


126
Although rumen fermentation dynamics were shown not to change markedly with
treatment, this study clearly demonstrates that cows fed a TMR containing ~ 8% citrus pulp
and supplemented with monensin improved their energy status indicated by lower NEFA and
BHB and higher glucose concentrations in blood. Although monensin boluses were inserted at
dry-off (50 to 70 d BEP), monensin still had positive effects on energy status of postpartum
dairy cows at 10 days of lactation, and during the entire period between dry-off and 21 d pp,
confirming the duration of the bolus of 95 days.
It is concluded that transition cows under the influence of monensin and fed a TMR
containing citrus pulp had lower levels of BHB, and NEFA and higher levels of glucose during
the entire experimental period and within 6 h after feeding. Body condition score at calving
was significantly affected by monensin in multiparous cows. Minor changes in rumen
fermentation dynamics were detected under the present experimental conditions.


142
At calving, the calf was immediately separated from the dam. If the cow needed calving
assistance she was moved to a maternity bam. Cows were processed within 12 h postpartum on a
routine basis, which included recording BCS, udder score (for edema presentation), reproductive
tract status (trauma or lacerations), and whether the cow was suspected of having retained fetal
membranes (RFM). If cows developed either RFM or milk fever they were treated and remained in
the hospital bam until recovery. After calving processing, cows were moved to a postpartum lot and
fed a diet higher in forage NDF (Tables 5.1 and 5.2). Any cow which had a decrease in milk yield
was moved from the milking herd and sent to the hospital bam to be examined and treated as
needed.
Beginning 60 days postpartum, cows received bST (Posilac, 500 mg sometribove zinc,
subcutaneously; Monsanto, St Louis, Missouri) every 14 days during the entire lactation.
Experimental Protocol
During July to August 2001, 60 cows dried-off 50 to 70 days BEP were randomly
assigned either a treatment or a control group. Treated group (n=30) received orally a capsule of
monensin (releasing 300 mg of monesin daily for 95 days, CRC Rumensin, ELANCO Animal
Health, Guelph, ON, Canada). Control cows (no capsule, n=30) were randomly matched by
parity.
The number of animals per treatment was calculated expecting a reduction in the
concentrations of BHB from 1000 to 850 52 pmol/L at 14 days postpartum (95% confidence, 80%
of power).
Body Condition Score and Milk Yield
Body condition score was evaluated using a standard methodology with an scale 1 to 5
(Ferguson et al., 1994). Body condition score was taken at assignment (dry-off) and at calving. Milk
yield was assessed using the first 10 Dairy Herd Improvement Association test day evaluations.


CHAPTER 6
EFFECT OF A MONENSIN-CONTROLLED RELEASE CAPSULE ON MILK AND
PLASMA B-HYDROXYBUTYRATE CONCENTRATIONS IN FLORIDA HOLSTEIN
TRANSITION COWS
Introduction
Ketosis is a common calving-related disorder affecting dairy cows during the transition
period, which is defined as the three weeks before and after parturition (Grummer, 1995;
Duffield et al., 1998a; 1998b). Ketosis may be expressed either as a clinical or subclinical
condition. Its subclinical expression is characterized by B-hydroxy butyrate (BHB) levels, >
1000 to 1400 pmol/L (Duffield, 2000; Geishauser et al., 2000). The highest incidence of
subclinical ketosis occurs within the first 2 to 3 weeks of lactation (Duffield, 2000);
therefore a monitoring program, testing all cows during the first two weeks of lactation may
be beneficial (Geishauser et al., 2001). The Ketolac test (Nagoya, Japan) is designed for
use in milk and has the sensitive to detect subclinical ketosis; therefore it is a practical tool
for use in a routine monitoring program to detect subclinical ketosis in early postpartum
dairy cows (Geishauser et al., 2000). If prevention is not considered during the transition
period, cows will be at higher risk of developing ketosis (Duffield et al., 1998b; Geishauser et
al., 2001).
Monensin is an ionophore that affects rumen fermentation (Van Maanen et al.,
1978), increasing propionic acid production with a concurrent decrease in the molar
proportion of acetate and butyrate (Richardson et al., 1976). As a result, monensin has been
used to prevent ketosis and related disorders in dairy cattle (Duffield et al., 1998a; 1999b;
Green et al., 1999; Duffield et al., 2003).
155


LIST OF FIGURES
Figure page
2-1. Pathways of glucose, amino acids and fatty acids
metabolism intersect at the citric acid cycle 58
2-2. Estimated energy density required in diets of transition
cows to meet requirements for maintenance and gestation 79
2-3. Estimated percentage crude protein required in diets of transition
cows to meet requirements for maintenance and gestation 81
3-1. Body Condition Score by treatments at assignment and at
calving 110
3-2. Daily milk yield up to 20 dpp in first lactation cows by
treatment Ill
3-3. Daily milk yield up-to 20 d pp in third lactation cows
or older by treatment 112
3-4. Test day milk yield by parity 1 and 2 ,113
3-5. Test day milk yield by parity 3 or more 114
3-6. Test day Fat % by parity 3 or more 115
3-7. Test day Protein % by parity 3 or more 116
3-8. Survival curve for risk of non pregnancy 117
4-1. BCS at dry-off and at calving in treated and control
group by parity 135
4-2. Glucose, BHB, NEFA concentration in monensin and
control primiparous cows by time after feeding at 10 d pp 136
4-3. Glucose, BHB, NEFA concentration in monensin and
control multiparous cows by time after feeding at 10 d pp 137
x


40
varied between 1.5 to 2.8 kg/day during the first 2 weeks after the diagnosis. However the
lactational incidence of lameness was as low as 2.1% over a population of 23,416 cows.
Lameness has been described more likely to occur during the first 60 to 90 days of
lactation (Rowlands et al., 1985). Grhn et al. (1990a) described a lactational incidence of
lameness of 1.9%, and the median day postpartum of diagnosis of the lameness was 65
days.
Although lameness has not been strongly related with calving-related disorders it
has been considered as a risk factor for some postpartum diseases in dairy cattle. Peeler et
al. (1994) found that cows that experienced dystocia were 1.47 times more likely to
become lame. Furthermore, lame cows were 1.45 times and 1.42 times more likely to
develop mastitis and anestrus than normal cows, respectively. In a logistic regression
model for different periparturient diseases, lame cows were 6.1 times more likely to
develop early metritis than non-lame cows. Finally, in a Florida study, lame cows within
the first 30 days postpartum were 2.63 times more likely to develop ovarian cysts and 0.43
times less likely to become pregnant than non-lame parity-matched cows (Melendez et al.,
2003a).
Culling
Culling is defined as the removal of animals from a dairy production enterprise.
Culling provides the opportunity to progress in genetic improvement, but it can also
represent a substantial loss to the producer (Radostits et al., 1994). Dairy herds can be
expected to cull between 20 and 35% of their cows each year. Genetic progress can be
also increased if the generation interval is shortened, reproductive performance is high,
and calf mortality is low (Radostits et al., 1994).


77
Neither monensin bolus nor premix had an effect on ruminal pH and VFA concentrations
under conditions of subacute rumen acidosis.
Since monesin is not allowed in lactating dairy cattle in the United States, only a
few studies have been conducted in this country. These studies have been under
supervision of the Food and Drug Administration. One of these studies was conducted to
evaluate the effects of monensin on nitrogen metabolism and the Cornell Net
Carbohydrate and Protein System (CNCPS) in cows fed fresh forage diets. Thirty Holstein
cows in mid lactation (8 with ruminal fistulas) where gradually introduced to a fresh
forage diet. Fifteen cows each were allocated to a control and a treatment group that
received 350 mg/cow per day of monensin in the P.M. concentrate feeding. A 7 day fecal
and urine collection period and a 3 day rumen sampling period were conducted with the
fistulated cows. Monensin increased milk production by 1.85 kg/d. Ruminal ammonia and
the acetate to propionate ratio decreased with the addition of monensin. Monensin
decreased fecal N output, and increased apparent N digestibility by 5.4%. It was
concluded that monensin has the potential to increase the efficiency of N utilization in
dairy cows fed fresh forage and to decrease fecal N excretion. The results suggest that
monensin spared amino acids from wasteful degradation in the rumen (Ruiz et al., 2001).
Transition Cow Feed Management
Feeding a cow during the transition period is a challenge due to the nutritional and
physiological changes that occur during this period (Grummer, 1999). Just recently
nutrient requirements for transition dairy cows have been defined (NRC, 2001). This
information was possible based on research that has been abundant during the past five
years. There have been tremendous changes in how to approach dry cow nutrition,
particularly in the areas of dry matter intake, protein and energy requirements, metabolic


188
Sharpe, K. L., H. Eiler, F. M. Hopkins. 1990. Changes in the proportion of type I and type III
collagen in the developing and retained bovine placentome. Biol. Reprod 43:229-
Shaver, R. D. 1997. Nutritional risk factors in the etiology of left displaced abomasum in
dairy cows: a review. J. Dairy Sci. 80:2449-2453.
She, P., A. R. Hippen, J. W. Young, G. L. Lindberg, D. C. Beitz, L. F. Richardson, and R. W.
Tucker. 1999. Metabolic responses of lactating dairy cows to 14-day intravenous infusions of
glucagon. J. Dairy Sci. 82:1118-1127.
Shearer, J. K. 1998. Lameness of dairy cattle: consequences and causes. The Bovine
Practitioner 32:79-85.
Shearer, J. K., and J. B. Elliot. 1998. Papillomatous digital dermatitis: treatment and control
strategies: part 1. Compend. Contin. Educ. Prac. Vet. 20:S 158-S165.
Shpigel, N. Y., R. Chen, Y. Avidar. 1996. Use of corticosteroids alone or combined with
glucose to treat ketosis in dairy cows. J. Amer. Vet. Med. Assoc. 208:1702-1704.
Simpson, R.B., C.C. Chase, A.C. Hammond, MJ. Williams, and T.A. Olson. 1998. Average
daily gain, blood metabolites, and body composition at first conception in Hereford, Senepol,
and reciprocal crossbred heifers on two levels of winter nutrition and two summer grazing
treatments. J. Anim. Sci. 76:396-403.
Smith, B. I., G. A. Donovan. C. Risco, R. Littell, C. Young, L. H. Stanker, and J. Elliot. 1998.
Comparison of various antibiotic treatments for cows diagnosed with toxic puerperal metritis.
J. Dairy Sci. 81:1555-1562.
Starnes, S.R., J.W. Spears, M.A. Froetschel, and W.J. Croom. 1984. Influence of monensin
and lasalocid on mineral metabolism and ruminal urease activity in steers. J. Nutr. 114:518-
525.
Stephenson, K.A., I.J. Lean, M.L. Hyde, M.A. Curtis, J.K. Garvin, and L.B. Lowe. 1997.
Effects of monensin on the metabolism of periparturient dairy cows. J. Dairy Sci. 80:830-837.
Stevenson, J. S., and E. P. Call. 1988. Reproductive disorders in the periparturient dairy cow.
J. Dairy Sci. 71:2572-2583.
Stewart, C.S., H.J. Flint, and M.P. Bryant. 1997. The rumen bacteria. Pages 10-72 in The
rumen microbial ecosystem. Second edition. P.N. Hobson and C.S. Stewart. Blackie
Academic & Professional.
Stock, R.A., S. B. Laudert, W. W. Stroup, E. M. Larson, J. C. Parrott, and R. A. Britton. 1995.
Effect of monensin and monensin and tylosin combination on feed intake variation of feedlot
steers. J. Anim Sci. 73: 39-44.


82
Minerals and Vitamins
Minerals are essential for production, reproduction and life of the animals. The
daily dietary requirement is dependent on the amount of dietary mineral that is absorbed
into the tissues (Goff, 2000). The requirement of the cow can be described by the dietary
requirement equal to maintenance plus pregnancy plus growth plus lactation divided by
the absorption co-efficient. In this equation, the upper portion can be obtained from NRC
(2001) or other table requirements. However the real challenge is to determine the co
efficient of absorption for the minerals.
Coefficient of absorption for Ca was 0.38 in NRC (1989) and 0.45 in NRC (1978).
ARC (1980) determined a co-efficient of 0.68. Maybe a single coefficient is not
appropriate. Availability of Ca in forages, concentrates or mineral supplements is different
(Goff, 2000).
Coefficient of absorption for P was 0.50 in NRC (1989) and 0.65 in NRC (1978).
Rest of the world use 0.60 to 0.75.
The average coefficient of absorption for Mg from a wide variety of natural
feedstuffs fed to ruminants averaged 0.294 with a standard deviation of 0.135. The co
efficient of absorption for Mg from inorganic sources should be 0.50 based on Mg oxide.
The coefficient for Mg absorption should be decreased when K is high in the diet (Goff,
2000)
Dietary Cl is absorbed with at least 80% and closer to 100% efficiency. Often Cl
anions accompany the movement of Na cations (Goff, 2000). Plants contain only small
amount of Na. Salt needs to be added to the diet, otherwise cows produce less milk.
Animals can tolerate very high levels of salt in the diet if water is provided and kidneys
are functioning. High dietary NaCl will reduce feed intake in animals (Goff, 2000).


72
cows. Blood urea nitrogen was elevated in the treatment group and NEFA were elevated
in the 2nd and 3rd month after treatment. There was no significant effect on blood
concentrations of glucose, albumin, or BHB, except in the 4th month where BHB was
higher in the treated group (1.75 mmol/L) than control group (1.35 mmol/L) (Hayes et al.,
1996). According to Duffield (2000), these levels are indicative of subclinical ketosis
(BHB >1.0 mmol/L), which might suggest that cows under those conditions were exposed
to marked energy imbalances.
In Italy, Ramanzin et al. (1997) using 4 primiparous Holstein cows in a Latin
square design with two forage to concentrate ratios (70:30 and 50:50, respectively) and
two concentrations of powder sodium monensin (0 and 300 mg/d/cow, offered
postpartum) determined that monensin tended to depress feed intake and milk fat content
without affecting milk production. Ruminal propionate percentage was increased more by
the addition of monensin to the low forage diet that to the high-forage diet. Serum urea
and NEFA concentrations tended to decrease when monensin was added to the low forage
diet.
In England, Phipps et al. (2000) conducted two experiments to examine the effect
of monensin on feed intake and milk production in Holstein-Friesian cows. In experiment
1, they used 60 lactating cows during weeks 7 to 26 of lactation. There were 4 treatment
groups receiving a 1 kg/d supplement containing 0, 150, 300 or 450 mg of monensin. Dry
matter intake was not different among groups, however milk production was higher in the
groups receiving 150, 300 and 450 mg/d of monensin. Percentage, but not absolute
content (g/d), of fat and protein were lower in the treated groups than in the control group.
In the second experiment, they used 69 multiparous and 29 primiparous Holstein cows
that received either 0 or 300 mg/d of monensin. Cows were evaluated during 2 lactations.


BIBLIOGRAPHICAL SKETCH
Dr. Pedro Melendez was bom in Santiago of Chile on January 6th of 1966. His
parents, Oscar and Eliana, were employees with no high level of education. With their
support and encouragement, they tried to offer the best education for their two sons,
Oscar and Pedro. Dr. Melendez grew up in Santiago and attended the San Marcos
Catholic School accomplishing his elementary, middle and high school levels. Since
middle school Dr. Melendez was interested in areas of biology and life sciences. He
always was impressed by the Jacques Couqsteau adventure series. In high school, he had
the desire to be a marine biologist, but when applying to the Chilean University system,
his mother suffered a car accident, which changed Dr. Melendez and his familys life for
ever. He was unable to be a marine biologist, but he started the veterinary medicine
career at the University of Chile. While his mother was recovering on bed for two years,
Dr. Melendez tried to do the best at the University. During the second year of his
veterinary education he discovered cows. One of his big influences was Dr. Mario
Duchens (actually one of his best friends) who introduced Dr. Melendez to the large
animal world, participating in several internships and summer jobs related to cattle. In
1990, Dr. Melendez received his degree in veterinary sciences. While he was working in
the private practice he helped in several courses offered by the veterinary school. He then
started to enjoy the academia and research. Thus, in 1992 he obtained a position as
instmctor at the Department of Animal Sciences, at the Faculty of Veterinary Medicine,
192


156
Monensin is not allowed for use in lactating dairy cattle in the United States. The
use of a monensin-controlled release capsule in Canada and Australia has been
demonstrated to be valuable in the prevention of calving-related disorders, primarily
ketosis, in transition dairy cows (Stephenson et al., 1997; Duffield et al., 1998b, Duffield et
ah, 1999b; Green et ah, 1999; Mutsvangwa et ah, 2002).
Citrus pulp is a common energy concentrate by-product feed used in dairy cattle in
Florida and other states (Arthington et ah, 2002). It contains a high proportion of pectin,
which is not digested by mammalian enzymes, but can be rapidly fermented by ruminal
microorganisms (Hall, 1997). Since pectin-fermenting bacteria are gram-negative
monensin-resistant bacteria (Nagaraja et ah, 1997; Stewart et ah, 1997), it is hypothesized
that dairy cows fed monensin and diets containing citrus pulp should not modify extensively
their pectin digestion and still monensin would affect positively energy status in Florida
transition dairy cows fed citrus pulp-based diets. Therefore, the objectives of this study
were to determine the effect of a monensin-controlled release capsule applied at dry-off on the
proportion of cows with concentrations of milk BHB > 200 pmol/L and to establish the
concentration of serum BHB at 14 d postpartum in Florida dairy cows fed citrus pulp and its
association with BHB concentrations in milk.
Materials and Methods
Cows and Herd Management
The study was conducted on a commercial Florida dairy farm with 3600 milking cows,
and milk rolling herd average of 10,700 kg. Most lactating cows were housed in a dry-lot
system and fed the same total mixed ration (TMR) three times a day, except postpartum
transition cows which received a diet higher in forage NDF. Cows were dried-off between 50 to
70 days before expected parturition (BEP) and maintained in a dry-lot (far-off cows) until 21


4-4. Glucose, BHB, NEFA concentration in monensin and control
primiparous cows by time between dry-off and 21 d pp 138
4-5. Glucose, BHB, NEFA concentration in monensin and control
multiparous cows by time between dry-off and 21dpp 139
5-1. Body Condition Score by treatments and by parity at assignment
and at calving 150
5 -2. Test day milk production by treatment in primiparous cows 151
5 -3. Test day milk production by treatment in multiparous cows 152
5-4. Glucose, BHB, NEFA concentration in monensin and control
primiparous cows by time between dry-off and 21 d pp 153
5-5. Glucose, BHB, NEFA concentration in monensin and control
multiparous cows by time between dry-off and 21 d pp 154
6-1. ROC curve for milk BHB for the detection of subclinical
ketosis under a gold standard of serum BHB >1.0 mmol/L 168
XI


20
membranes and lesions can be produced in the upper digestive tract, rumen and
abomasum (Goff 1999a). Calcium absorption in the rumen is less efficient than in the
intestine because the volume of fluid in rumen will rapidly dilute the Ca concentration to a
value less than 6 mmol/L required for passive absorption. Pastes and gels reduce the
amount of Ca likely to bypass the rumen. Thus, in general, more Ca must be administered
by gel or paste to obtain the same rise in plasma Ca as is achieved by an oral Ca drench.
However, drenches present the disadvantage that some cows will aspirate the solution,
which can lead to severe aspiration pneumonia (Goff, 1999a). Calcium propionate is
effective and less irritating to tissues than is calcium chloride. It does not induce metabolic
acidosis, so larger amounts of Ca can be given. Furthermore, it supplies the cow with a
gluconeogenic precursor (propionate) (Goff, 1999a). Oral treatments increase Ca within
30 to 60 minutes of administration, and plasma Ca concentrations remain elevated for
about 6 hours. Calcium chloride acts a little faster, but calcium propionate may act a little
longer (Goff, 1999a).
For many years, the traditional method of preventing MF in dairy cows was the
restriction of dietary intake of Ca during the prepartum period. Ca diets with < 15-20 g of
Ca/d fed during the last 10 days of gestation, followed by a postpartum diet that is high in
Ca have been recommended. These diets will greatly reduce the risk of MF (Horst et al.,
1994; Joyce et ah, 1997; Vagnoni and Oetzel, 1998 ; Oetzel and Goff, 1999). According to
NRC (2001), the requirements of Ca for a 680 kg mature dry pregnant cow (maintenance
plus last 2 month of gestation) is 0.45% dry matter basis.
Oral and intramuscular doses of vitamin D3 have also prevented MF successfully.
However, repeated treatments may lead to toxicity (Jorgensen, 1974; Markusfeld, 1989).
Parathyriod hormone has been also reported to prevent parturient paresis in dairy cows.


95
between the 9th and 12th rib spaces, with or without colic pain. Culling rate was defined
as number of cows that left the herd for any reason until 410 d postpartum divided by
total number of cows that were enrolled in the trial. Other outcome variables were daily
milk yield up to 20 days post partum and milk production, milk fat and protein content
at DHLA. test days during the entire lactation. Mature Equivalent 305 milk production
was also recorded. Body condition score was performed at assignment, and at calving
according to a standard methodology by the same person (Ferguson et al., 1994).
Reproductive parameters considered to evaluate fertility were conception rate at
first service (CRIS), overall pregnancy rate (PR), calving to conception interval, (CCI)
and number of services per conception (SC). Conception rate at first service was defined
as the number of cows pregnant at their first service divided by number of cows
inseminated the first time after calving. Overall PR was defined as the number of cows
which became pregnant up to 410 d of lactation divided by the number of cows
inseminated at least once during that time. Overall CCI was defined as the average days
between calving and the breeding date after which cows were confirmed pregnant.
Services per conception were defined as the average number of inseminations in cows
that conceived.
Statistical Analysis
The null hypothesis was that there is no difference in the incidence of CRD and
other outcomes between treatment groups. Incidences were analyzed using logistic
regression modeling through a backward model-selection procedure. To determine the
degree of association between the risk factors and outcome variables, odds ratio (OR)
and 95% confidence intervals were calculated. Treatments were forced to remain in the
final models. Milk yield was analyzed constructing a mixed model for repeated


111
Figure 3-2. Daily milk yield up to 20 d pp in first and second lactation cows by treatment.
* P < 0.05


106
Table 3-2. Nutrient content of dry cow far-off, dry-cow transition and lactating transition
diets
Nutrient
Dry cow
far-off
Dry cow
transition
Lactating
transition
CP(%DM) 1
15.52
17.91
18.60
UndegP(%CP)2
26.38
35.31
30.34
DegP(%CP)2
73.62
64.69
69.66
SolP(%CP)2
-
36.72
39.77
NEL(Mcal/kg)3
ADF(%DM) 1
0.84
1.69
1.69
24.54
25.13
23.66
NDF(%DM) 1
36.04
36.20
34.63
NFC(%DM)2
32.94
31.55
34.13
Starch(%DM)2
11.05
14.96
14.90
Lipid (%DM)1
2.85
6.20
2.36
Ca(%DM)1
P(%DM)1
0.74
1.27
1.10
0.30
0.35
0.46
Mg(%DM) 1
0.32
0.36
0.36
K(%DM)1
1.28
1.10
1.46
Na(%DM)1
0.09
0.09
0.58
C1(%DM) 1
0.18
0.43
0.48
S(%DM)1
0.24
0.40
0.22
Forage in diet (%DM)
60.21
48.10
50.23
Cation-anion (meq/kg DM) 4
165.90
-51.7
351.90
Laboratory nutritional analysis
Values from feed composition tables
3 From formulas after laboratory analysis
4 From formula (Na+ + K+) (CF + S')


108
Table 3-4. Summary of logistic regression modeling for CRD1
Model
Coefficients
Dependent
Independent
or(2)
95%CI(3)
p-value
Dystocia
Monensin yes
2.10
1.39-3.17
0.0004
Parity primiparous
0.65
0.43-0.99
0.049
Metritis
Monensin yes
0.80
0.70- 0.88
<0.0001
Parity primiparous
5.65
3.25-9.82
<0.0001
Dystocia yes
19.2
10.7-38.0
<0.0001
(1) Calving-related disorders
(2) Odds Ratio
(3) 95% Confidence Intervals (Odds Ratio)


19
calcium lactate, which tend to be less injurious to tissues than other forms of calcium
(Goff, 1999a).
A variety of oral calcium salt preparations are available for cattle (Oetzel and Goff,
1999). Oral Ca supplement must be readily soluble in water (digestive fluids) to reach the
minimum amount for passive transport (~ 6 mmol/L) (Goff, 1999a). About 4 g of calcium
will be absorbed and enter the bloodstream of a cow given an oral solution containing 50 g
of calcium chloride (Oetzel and Goff, 1999). Calcium chloride is the most soluble of the
Ca salts; calcium propionate, calcium formate, calcium acetate, calcium gluconate and
calcium lactate are also soluble enough. Calcium hydroxide, calcium oxide and calcium
carbonate are relatively insoluble and unsuitable for treating hypocalcemia. Ca that is not
absorbed by passive diffusion is still available for absorption by active Ca transport in the
small intestine, but this absorption is not rapid enough to be of aid in the treatment of
hypocalcemia (Goff, 1999a). Oral Ca products typically contain between 25 and 100 g of
Ca (Oetzel and Goff, 1999). Calcium chloride and calcium propionate are the most
common products used in the treatment and prevention of hypocalcemia in cattle (Goff,
1999a). Oral administration of 50 g of calcium from calcium chloride as a drench in 250
mL of water raises plasma Ca concentrations to the same extent as 4 g of Ca as calcium
chloride administered intravenously. Conversely, 100 g of Ca orally is equivalent to
between 8 and 10 g of calcium administered intravenously. Calcium chloride increases
plasma Ca better than calcium propionate (Goff and Horst, 1993). Calcium chloride is
slightly more effective and takes up less volume than calcium propionate; it also is an
acidifying agent. This mild metabolic acidosis can help enhance Ca homeostasis.
However, severe metabolic acidosis can be produced when repeated treatments of calcium
chloride are administered. Furthermore, calcium chloride is very irritating to mucous


183
Muller, L. D.1992. Feeding management strategies. Pages 326-335 In Large Dairy Herd
Management. H. H. Van Horn and C. J. Wilcox. American Dairy Science Association, 301
West Clark St., Champaign, IL 61820.
Miettinen, H., and P. Huhtanen. 1996. Effects of the ratio of ruminal propionate to butyrate on
milk yield and blood metabolites in dairy cows. J. Dairy Sci. 79:851-861.
Milian-Suazo, F., H. N. Erb, and R. D. Smith. 1988. Descriptive epidemiology of culling in
dairy cows from 34 herds in New York state. Prev. Vet. Med. 6:243-251.
Miller, B. J., and J. R. Lodge. 1984. Postpartum oxytocin treatment of retained placenta.
Theriogenology 22:385-
Miller, G., and C. Dorn. 1990. Cost of dairy cattle diseases to producers in Ohio. Prev. Vet.
Med. 8:171-182
Minor, D. J., R. R. Grummer, R. D. Shaver, and S. L. Trower. 1996. Effect of niacin, and
nonfiber carbohydrate on the metabolic status during the transition period and lactation
performance. J. Dairy Sci. 79(Suppl. 1): 199
Minor, D. J., S. L. Trower, and B. D. Strang. 1998. Effects of nonfiber carbohydrate and
niacin on periparturient metabolic status and lactation of dairy cows. J. Dairy Sci. 81: 189-
200.
Mutsvangwa, T., J.P. Walton, J.C. Plaizier, T.F. Duffield, R. Bagg, P. Dick, G. Vessie, and
B.W. McBride. 2002. Effects of a monensin controlled-release capsule or premix on
attenuation of subacute ruminal acidosis in dairy cows. J. Dairy Sci. 85:3454-3461.
Nagaraja, T.G., C.J. Newbold, C.J. Van Nevel, and D.I. Demeyer. 1997. Manipulation of
ruminal fermentation. Pages 523-632 in The rumen microbial ecosystem. Second edition. P.N.
Hobson and C.S. Stewart. Blackie Academic & Professional.
National Research Council. 1978. Nutrient requirements of dairy cattle, 5th revised edition.
Washington, D.C.: National Academy of Sciences
National Research Council. 1989. Nutrient requirements of dairy cattle, 6th revised edition.
Washington, D.C.: National Academy Press.
National Research Council. 2001. Nutrient requirements of dairy cattle, 7,h revised edition.
Washington, D.C.: National Academy Press.
Nelson, D. L. and M. M. Cox. 2000. Oxidation of fatty acids. Pages 598-622. In Lehninger
Principles of biochemistry. Third edition. Worth Publishers, New York, NY 10010.


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Markusfeld, O. 1989. The evaluation of a routine treatment with la-hydroxyvitamin D3 for
the prevention of bovine parturient paresis. Prev. Vet. Med. 7:1-9.
Markusfeld, O., N. Galon, and E. Ezra. 1997. Body condition score, health, yield and fertility
in dairy cows. Vet. Rec. 141:67-72.
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Therapy 4. Food Animal practice. J. Howard and R. Smith. W.B. Saunders Company.
Massey, C. D., C. Wang, G. A. Donovan, and D. Beede. 1993. Hypocalcemia at parturition as
a risk factor for left displacement of the abomasum in dairy cows. J. Amer. Vet. Med. Assoc
203:852-853.
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Current status and future outlook. J. Dairy Sci. 84 (E. Suppl.):E194-E203.
Meinert, R.A., C.M.J. Yang, A.J. Heinrichs, and G.A. Varga. 1992. Effect of monensin on
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and chewing activity in Holstein cows. J. Dairy Sci. 86 (Suppl. 1):W227 (Abstr.)
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ovarian cysts and fertility in lactating dairy cows. Theriogenology, 59: 927-937.
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responses of transition Holstein cows fed anionic salts and supplemented at calving with
calcium and energy. J. Dairy Sci. 85:1085-1092.
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supplements on calving-related disorders, fertility and milk yield during the transition period
in cows fed anionic diets. Theriogenology 60:843-854.
Melendez, P, J. McHale, J. Bartolom, A. Sozzi, S. Lanhart, A. Donovan. 2003c. The effect of
pgf2a at day 8 post partum on uterine involution and acute response proteins in Holstein cows
with acute puerperal metritis. Tenth International Symposium on Veterinary Epidemiology
and Economics (submitted).
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particle size at initial feeding and the weigh back and chewing activity in dairy cattle. The
Bovine Practitioner 36: 66-70.
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Association, 301 West Clark St., Champaign, IL 61820.


33
decreased as lactation number increased. At the level of the herd, factors associated
positively with risk of LDA were Predicted Transmitted Ability (PTA) for milk
production, BCS, winter and summer seasons, and precalving rations containing energy
densities > 1.65 Meal of Enl/kg of DM. Feed bunk management considering bunk space,
feed availability and freshness was associated negatively with the risk of LDA.
Nutrition has been implicated as one of the most important risk factors in the
etiology of LDA. The transition period from two weeks prepartum through two to four wk
postpartum is the major risk period for the occurrence of LDA (Shaver, 1997). This might
be explained by a low feed consumption during the transition period where rumen fill is
decreased. The decline of dry matter intake is about 35% over the final week prepartum
(Bertics et al., 1992). Furthermore, as pregnancy progresses, the growing uterus occupies
an increasing amount of the abdominal cavity. This forces the abomasum forward and
slightly to the left side of the cow. After calving, if the smaller rumen does not move into
its normal position on the left ventral floor of the abdomen, the abomasum is able to slide
under it (Goff and Horst, 1997b).
In Florida, Massey et ah, (1993) reported that hypocalcemic cows at parturition
(total serum calcium < 7.9 mg/dl) were 4.9 times more likely to develop LDA than
normocalcemic cows. Hypocalcemia is known to cause abomasal atony. Abomasal atony
is an absolute prerequisite to LDA (Fecteau et ah, 1999). Strategies to prevent
hypocalcemia at parturition may be useful for the prevention of LDA (Shaver, 1997).
Ketosis is the strongest identified risk factor associated with LDA. Curtis et ah,
(1985) reported that cows with uncomplicated ketosis were 11.9 times more likely to
develop an LDA than normal cows. Correa et ah, (1993) found that cows with ketosis
were 13.8 times more likely to develop an LDA than normal cows. Melendez et ah


27
weeks after calving. The condition is characterized by an abnormal cervical discharge,
vaginal discharge, or both (Lewis, 1997; Youngquist and Shore, 1997; Kelton et al.,
1998). The condition may be local or systemic. A condition called toxic or gangrenous
metritis, that occurs almost exclusively in the puerperal period and is often associated with
clostridial infections, is characterized by foul-smelling, watery uterine discharge, severe
drop in milk production and systemic symptomatology (Gilbert and Schwark, 1992; Olson
et ah, 1986; Smith et ah, 1998).
Several microbiological risk factors have been associated with uterine infections.
The uterus has an anaerobic environment. Cows with RFM present a higher incidence of
coliforms, and other environmental bacteria, Clostridia, Archenobacter pyogenes, and
gram-negative anaerobes than normal cows (Olson et ah, 1986). Recent studies have
confirmed that A. pyogenes, either alone or in combination with anaerobic bacteria
(Fusobacterium necrophorum and Bacteroides spp.) can act to induce uterine infections in
cows during the puerperal or luteal phase (Del Vecchio et ah, 1992). These bacteria
produce postpuerperal metritis or a more delayed typical endometritis (Olson et ah, 1986;
Lewis, 1997; Youngquist and Shore, 1997). Postpuerperal metritis eventually become a
chronic problem. Affected cattle do not present a systemic disease but they may have a
mucopurulent, fetid vulvar discharge.
The incidence of uterine infections varies considerably among studies, and the
average incidence is not an especially meaningful statistics (Lewis, 1997). This variation
is due to poorly described diagnostic methods and lack of good definition of a case. Based
on 43 citations from 1979 to 1995, the frequency of metritis ranged from 2.2% to 37.3%.
The median was 10.1% (Kelton et ah, 1998). Grhn et ah, (1998) determined an incidence
of metritis of 4.2% in 7523 dairy cows in New York state.


17
parathyroid glands and can also alter the responsiveness of tissues to PTH. High dietary
potassium reduces ruminal magnesium absorption (Oetzel and Goff, 1999). Increase
dietary P intake (> 80 mg/day) increases the P in blood (~ 8 mg/dL), which has a direct
inhibitory effect on the renal enzymes that catalyze production of l,25(OH)2D. This
reduced production of l,25(OH)2D further reduces intestinal Ca absorption mechanism
prepartum (Horst et al., 1994; Oetzel and Goff, 1999).
One of the most important determinants of MF risk is the acid-base status of the
animal at the time of parturition. Metabolic alkalosis appears to alter the physiologic
activity of PTH so that bone resorption and production of l,25(OH)2D are impaired, thus
reducing the ability of the animal to successfully adjust to increased calcium demands
(Oetzel and Goff, 1999).
Milk fever is confirmed by low serum calcium concentrations. Clinical signs may
begin as total blood calcium values fall below 7.5 mg/dL (< 1.8 mmol/L). However more
than half of all mature dairy cows will have total blood calcium concentrations below 7.5
mg/dL (< 1.8 mmol/L) after calving without any evidence of clinical signs (Goff, 1999a;
Oetzel and Goff, 1999). Generally, cows that are recumbent and unable to rise as a result
of low blood Ca will have plasma Ca concentration less than 5 mg/dL, and some will be
down as low as 2 mg/dL; below this level is generally incompatible with life (Goff,
1999a). Milk fever presents three stages of clinical symptoms. Animals in stage I usually
have levels of 5.5 to 7.5 mg/dL of Ca. Animals in stage II typically have a total Ca
concentration of 3.5 to 6.5 mg/dL. Animals in stage III may have Ca levels as low as 1.0
mg/dL. Blood concentrations of P are typically below normal and magnesium
concentrations are usually high. The most important differential diagnosis is related to the
downer cow syndrome (Oetzel and Goff, 1999).


NEFA (mEq/L) Glucose (mmol / L) BHB (mmol/L)
138
Figure 4-4. Effect of monensin treatment on glucose, BHB, and NEFA
concentration in primiparous cows as a function of time


Milk fat (%)
115
Figure 3-6. Least squares means and SEM for test day Fat % by parity 3 or
greater. *P < 0.05, ** P<0.10.


67
Table 2-8. Susceptibility and resistance of ruminal bacteria to monensin.
Species
Gram
reaction
Cell wall
type
Monensin
susceptibility
Hydrogen and formic acid producers
Lachnospira multiparus
-
+
+ (0.38)a
Ruminococcus albus
-
+
+ (0.38)
Ruminococcus flavefaciens
-
+
+ (0.38)
Butyric acid producers
Butyrivibrio fibrisolvens
_
+
+ (0.38)
Eubacterium cellulosolvens
+
+
+ (0.38)
Eubacterium ruminantium
+
+
+ (0.38-1.5)
Lactic acid producers
Lactobacillus ruminis
+
+
+ (1.5-3.0)
Lactobacillus vitulinus
+
+
+ (0.38-1.5)
Streptococcus bovis
+
+
+ (0.38-12.0)
Ammonia producers
Clostridium aminophilum
+
NDb
ND
Clostridium sticklandii
+
+
ND
Peptostreptococcus anaerobius
+
+
ND
Succinic and propionic acid producers
Anaerovibrio lipolytica
-
-
- (>48.0)
Fibrobacter succinogenes
-
-
- (>20.0)
Megasphaera elsdenii
-
-
- (>48.0)
Prevotella ruminicola
-
-
- (>48.0)
Ruminobacter amylophilus
-
-
- (>48.0)
Selenomonas ruminantium
-
-
- (>48.0)
Succinimonas amylolytica
-
-
- (>48.0)
Succinivibrio dextrinosolvens
-
-
- (>48.0)
Methane producers
Methanobrevibacter ruminantium
NAC
NA
- (>20.0)
Methanobacterium formicium
NA
NA
- (>20.0)
Methanosarcina barkeri
NA
NA
- (>20.0)
Adapted from: Nagaraja, T.G., C.J. Newbold, CJ. Van Nevel, and D.I. Demeyer. 1997.
Manipulation of ruminal fermentation. Pages 523-632 in The rumen microbial ecosystem.
Second edition. P.N. Hobson and C.S. Stewart. Blackie Academic & Professional.
a Values in parenthesis are minimum inhibitory concentrations in pg ml"1
b Not determined
c Not applicable


62
Mitsuokella multiacidus: These gram-negative bacteria were previously regarded as
Bacteroides ruminicola. They produce lactate as a major fermentation product.
Megasphaera elsdeniv. These gram-negative cocci are mainly found in the rumen of
young animals. They ferment small sugars, glucose, fructose, and mostly lactate. This
bacterium is thought to play a major role in the production of branched-chain volatile
fatty acids. End products are butyrate, propionate, isobutyrate, valerate, C02 and some
H2.
Ruminococcus species: Represented mostly by two species involved in plant fiber
degradation in the rumen, R. albus and R. flavefaciens. They are gram-positive
cellulolytic bacteria. Major products are acetate, succinate, C02 and H2. They are
extremely sensitive to monensin.
Streptococcus bovis: Gram-positive cocci capable of very rapid growth. It is the most
rapidly acting amylolytic bacteria of the rumen. The major fermentation product is
lactate. Minor products are acetate, formate and C02. Lactate production increases by
the conditions that develop when large amounts of starch are fed and the rumen pH
falls.
Butyrivibrio fibrisolvens: Gram-negative, strictly anaerobic rods. Predominant bacteria
in the rumen of cows fed on a variety of different rations, ranging from alfalfa hay to
grain mixture. It is the major butyrate-producing species in the rumen, but they also
produce acetate, succinate, C02 and H2. They ferment cellulose and xylans.
Lachnospira multpara: Gram-positive curve rod degrading pectins. Major products
are formate, acetate, lactate, C02 and H2.


94
Experimental Protocol
During July to August 2001, 580 cows 50 to 70 days BEP were randomly
assigned to either a treatment or a control group. The treated group (n=290) received
orally a capsule of monensin (releasing 300 mg of monesin daily for 95 days, CRC
Rumensin, ELANCO Animal Health, Guelph, ON, Canada). Control cows (no capsule,
n=290) were randomly matched by parity (1, 2, 3 or greater).
The number of animals per treatment was calculated expecting a reduction in the
incidence of any calving-related disorders of 6% (95% confidence, 80% of power).
Outcome Variables and Case Definitions
Outcome variables were the incidence of dystocia, MF, RFM, metritis, clinical
ketosis, DA and culling rate. Incidence was defined as the number of specified new
cases during the entire lactation divided by the total number of cows in that group.
Dystocia was defined as human intervention (2 or more people) conducting forced
extraction of the calf for more than 5 minutes. Milk fever was defined as any downer
cow within 72 hours after parturition presenting off-feed, nervous symptoms, staggering,
varying degrees of unconsciousness and good response to intravenous calcium
treatment. Retained fetal membranes were defined as visible fetal membranes at the
vulva, vagina or uterus by vaginal examination more than 24 hours after calving.
Metritis was defined as any abnormal vaginal discharge or uterine content (foul
smelling) obtained by rectal palpation. Clinical ketosis was defined as decreased
appetite, decrease in milk production with elevated urine ketones as measured by a
commercial test (Ketostix, Bayer Corporation Elkhart, IN 46515). Displacement of
abomasum was defined as decreased appetite and milk yield accompanied by an audible
high pitched tympanic resonance elicited by percusin of the left or right abdominal wall


163
Table 6-1. Diet composition for far-off dry cow, transition dry cows and lactating
transition cows
Feed
Dry cows
far-off
Dry-cows
transition
Lactating
transition
Alfalfa hay
-
19.30
17.53
Coastal hay
-
4.76
-
Cottonseed whole
-
9.84
8.75
Com silage
44.90
24.06
28.50
Ryegrass silage
15.29
-
4.20
Com hominy
-
6.98
13.54
Citrus pulp
21.20
9.82
4.25
Soybean meal 48
17.20
3.44
5.49
Wet brewers grain
-
10.74
4.07
Lactating concentrate
-
-
7.03
Lactowhey 1
-
3.76
4.23
Metaxerol2
-
2.79
1.96
Springer minerals
-
4.51
-
Dry cow minerals
1.41
-
-
Lactating minerals
-
-
0.45
1 Ammoniated whey (61.5% CP)
2 Energy supplement based on sodium propionate, propylene glycol, dried whey and calcium
carbonate


13
Table 2-3 Continued
Author
Disease
Risk factors
Association
Collard et
Laminitis
Energy balance postpartum
Negative
al., 2000
Digestive
Energy balance postpartum
Negative
Bruun et
Metritis
Dystocia
Positive
al., 2002
Reproductive disease
Positive
Retained placenta
Positive
Schnier et
Mastits
Warm housing system
Positive
al,2002
Metritis
Cold housing system
Negative
Melendez
et al.,
2003a
Ovarian cysts
Lameness
Positive
Melendez
et al.,
Ketosis
Displacement of abomasums
Positive
2003b
Displacement of
Retained fetal membranes
Positive
abomasums
Ketosis
Positive
Parity
Positive
Metritis
Retained fetal membranes
Positive
Parity
Negative


9
incidence. Some exceptions are lameness (h2=0.16) and ketosis (h2= 0.39) and the
selection on conformation traits can help reduce the incidence of disease, although genetic
correlations are low (Van Dorp et al., 1998).
There are several studies that have described the relationship and risk factors
among calving-related disorders in dairy cattle (Table 2-3) (Curtis et al., 1984; Curtis et
al., 1985; Erb et al., 1985; Grohn et al., 1989; Erb and Grohn, 1988; Correa et al., 1990;
Correa et al., 1993; Braun et al., 2002; Melendez et al., 2003a; Melendez et al., 2003b).
Results in general have been consistent, but some weaknesses of these studies have been
the variability of the case definition. In an effort to homogenize criteria definitions, Kelton
et al. (1998) recommended some guidelines for recording and calculating selected clinical
diseases in dairy cattle (Table 2-4).
Calving-related disorders result in significant economic losses to dairy producers
through reduction in reproductive performance and milk yield during the subsequent
lactation, cost of treatments and increased culling (Curtis et al., 1984; Curtis et al., 1985;
Erb et al., 1985; Grohn et al., 1989; Erb and Grohn, 1988; Correa et al., 1990; Correa et
al., 1993; Braun et al., 2002; Risco and Melendez, 2002).
The most relevant calving-related disorders are milk fever or hypocalcemia,
retained fetal membranes, metritis, ketosis, displacement of the abomasum, mastitis and
lameness (Correa et al., 1993; Goff and Horst, 1997b; Risco and Melendez, 2002).
In Table 2-4 case definitions, economic losses, epidemiology and recommended
guidelines for recording and calculating selected calving-related disorders in dairy cattle
are reported.


69
Since pectin-fermenting bacteria (Prevotella spp, Fibrobacter succinogenes,
Succiniovibrio dextrinosolvens) are gram-negative monensin-resistant bacteria (Nagaraja
et al., 1997; Stewart et al., 1997), it is reasonable to assume that in dairy cows fed
monensin and diets rich in citrus pulp ruminal pectin fermentation should not be altered
significantly.
Monensin. Calving-Related Disorders. Reproductive and Productive Responses
Ionophores have been available for use in food animals for over 20 years,
however, limited information is available about their effects in lactating dairy cows
(Duffield, 1997). During the last 5 years, more information about the use of monensin in
dairy cattle has been developed. Mostly this research has been conducted in Canada and
Australia. Monensin is not allowed for use in lactating dairy cattle in the United States.
One of the earliest reports showing the positive effect of monensin on ketosis
demonstrated that monensin at a rate of 250 mg per cow per day, successfully controlled
an outbreak of ketosis in 18 Friesian cows (Rogers and Hope-Cawdery, 1980). In a well-
designed study conducted in Canada by Sauer et al (1989), cows of second lactation or
older were gradually introduced to a monensin-containing concentrate 1 week prepartum
and fed complete diets containing 0, 15 or 30 g monensin/ton of dry matter for 3 weeks
postpartum (low and high monensin). Acetate molar proportion decreased, while
propionate increased significantly between controls and the high monensin-treated group.
The acetate to propionate ratio was 2.49 and 1.46 in the control and high monensin
groups, respectively. Monensin also decreased the levels of serum BHB in cows in the
high monensin group compared with the control group (3.9 mg/dL vs 7.2 mg/dL,
respectively). Milk production was similar, but milk fat was lower in the high monensin
group than the control group (3.71% vs 4.12%, respectively).


ACKNOWLEDGMENTS
I wish to express my sincere gratitude to:
Dr. Art Donovan (my adviser and friend) for his excellent advice, support and
enthusiasm on my project and my entire graduate program at UF.
Dr. Jesse Goff, distinguished USDA researcher, for accepting to be part of my
committee, for his expertise and friendship.
Dr. Carlos Risco, Professor of Veterinary Medicine for his valuable comments
and help and for his friendship.
Dr. Ramon Littell, Professor of Statistics, IFAS, for accepting to be part of my
committee and for his valuable help on the statistical approach.
Dr. Louis Archbald, Professor of Veterinary Medicine and LACS graduate
coordinator, for accepting to be part of my committee and for his advice on my entire
program.
Dr. Charles Courtney and the Graduate School of the College of Veterinary
Medicine, UF.
The Department of Large Animal Clinical Science and Dr. Eleanor Green
Pharmacia Animal Health for their financial support.
Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile.
Mr. Don Bennink, Dr. Jeniffer McHale, Bobby, and the staff of North Florida
Holsteins Dairy Farm.
ill


25
et al., (1985) determined that cows suffering hypocalcemia were 4.2 times and 2.0 times
more likely to have dystocia and RFM, respectively.
A recently explored, but old concept related to RFM is the immunological aspect
involved in the pathogenesis of this reproductive condition. The old theory suggests that
fetal membranes must be recognized as "foreign" tissues and rejected by the immune
system after parturition to cause expulsion of fetal membranes (Gunnink, 1984).
Neutrophils isolated from cows that experienced RFM had significantly lower function
than cows without RFM, before calving and during the first 2 weeks postpartum. In
addition, Interleukin-8, a potent neutrophil chemoattractant, was lower at calving in cows
with RFM (5112 pg/ml) than cows without RFM (13411 pg/ml). These findings suggest
that neutrophil function around parturition is a determining factor for the development of
RFM in dairy cattle (Kimura et al., 2002).
Retained fetal membranes have been the major factor that predispose cattle to
metritis. The majority of affected cows show no serious clinical signs other than a
transient decrease in appetite and milk production. However, 20% to 25% of cows
affected by RFM develop moderate to severe metritis (Joosten et al., 1988).
The relative risk of cows with RFM developing metritis was 2.8 and the
attributable risk was 28% as compared with cows without RFM (Bartlett et al., 1986). In
several epidemiological studies using path analysis methodology cows with RFM have
been around 6.0 times more likely to develop metritis than normal cows (Correa at al.,
1993; Curtis et al., 1985; Erb et al., 1985). Overall, studies using path analysis and risk
assessment indicated consistently that dystocia, nutrition, metabolic disorders and mainly
RFM increased the likelihood that a cow would develop metritis (Lewis, 1997; Bruun et
al., 2002; Melendez et al., 2003b).


99
Unexpected and surprisingly was the higher incidence of dystocia experienced
by older cows treated with monensin than control cows. Although dystocia is also a
multifactorial condition, those providing calving assistance were blind to treatment and
used consistent with farm protocols. A logical explanation of this finding was that older
cows treated with monensin had a higher BCS at calving than controls (P < 0.01).
Therefore, fat deposition might have also occurred inside the pelvis or birth canal, which
might have contributed to calving difficulty due to a reduced intra-pelvic space. Higher
body condition score at calving has been associated with higher incidence of dystocia
(Hoffman et al., 1996; Chassagne et al., 1999). Another explanation for these findings
might be that treated cows might have had heavier calves at parturition (larger calf birth
weight) since glucose and amino acids are the major fuel supply of the developing fetus
in ruminants (Drackley et ah, 2001). Unlike Canadian studies where boluses were
inserted during the last 3 weeks of gestation (Duffield et ah, 1998a; 1998b; Green et ah,
1999), in the present study boluses were placed 50 to 70 days BEP, which might have
substantially increased glucose availability for the fetus. In beef cattle, calves from
heifers fed monensin had higher birth weights than those from control heifers (Clanton
et ah, 1981). Unfortunate under the study farm management conditions it was not
possible to weigh calves at birth.
The incidence of metritis in this study was considered in the upper normal values
reported by the literature (30 to 40%) (Kelton et ah, 1998). This high incidence might be
due to farm definition and might be an overestimation of the true incidence. In the
present study, abnormal vaginal discharge was used as a criterion. Other studies have
used a more precise definition considering body temperature, milk yield, and uterine
status at palpation in addition to vaginal discharge as their criteria (Smith et ah, 1998).


73
Cows evaluated during the second lactation also started to receive monensin 3 weeks
before expected parturion. Milk production was not different between groups in both
lactations, however percentage of fat and protein milk was lower in treated cows than
control cows in both lactations. The use of monensin before calving decreased the BHB
concentration in weeks 2 to 4 and 6 to 8 postcalving. Levels of BHB were between 380
and 524 pmol/L, values lower than the cut-off for subclinical ketosis (Duffield, 2000).
Monensin also reduced acetoacetate values in weeks 2 to 4 and increased glucose values
in weeks 2 to 4 and 6 to 8 postpartum. However, there was no difference in these
metabolites in weeks 10 to 12.
In another European study, Van der Werf et al. (1998) in the Netherlands
examined the efficacy of premix monensin on milk production and feed efficiency during
the first 20 weeks of lactation. In a first experiment they used 64 Holstein cows assigned
to 4 groups that received 0, 150, 300, or 450 mg/day of monensin mixed with concentrates
from 5 to 24 weeks postpartum. In a second experiment they utilized 58 Holstein and 22
Jersey cows that were allocated either to a control group or to a treatment group that
received 300 mg/day of monensin from 5 to 36 weeks postpartum during to consecutive
lactations. In trial 1 monensin caused a decrease in fat content for cows fed 450 mg/day.
Feed intake of treated cows tended to be lower than controls. Body weight gain was higher
for cows fed 450 mg/day of monensin. Acetoacetate, BHB and glucose did not differ
among groups. In trial 2, monensin increased milk production during the first lactation in
Holsteins, but not in Jersey cows. At day 56 of treatment a larger decrease in acetoacetate
and BHB and an increase in glucose were found for the treatment group. During the
second lactation acetoacetate and BHB were significantly lower and glucose was higher
for cows in the treatment group than controls. Productive responses were not different.


165
Table 6-3. Receiver-operating characteristic (ROC) analysis for serum BHB and Kappa
estimator (agreement evaluation) between milk BHB and serum BHB
Serum BHBA
concentration
mmol/L
Area under the curve
(%)
Lower-upper limit
Kappa estimator
milk BHB
> 200 pmol/L
>0.8
78.01
72.96-83.07
0.36
> 1.0
83.73
79.31-88.15
0.37
> 1.2
86.26
81.54-90.99
0.30


22
Diets fed prior to parturition that evoke an acidic response in the animal reduce
MF risk, whereas diets that evoke an alkaline response increase it. Thus, low DC AD diets
cause metabolic acidosis and reduce the risk of MF. A diet can have a low DC AD because
it is low in cations, high in anions, or a combination of both (Oetzel and Goff, 1999). As
DCAD decreases, H+ increases, HCO"3 decreases and pH decreases. These changes are
accompanied by a reduction in urinary HCO'3 excretion and urinary pH as compensatory
mechanisms. Furthermore, low DCAD prepartum can mitigate hypocalcemia via increased
urinary Ca reabsorption, serum ionized Ca, and responsiveness to Ca homeostatic
hormones (Block, 1994; Vagnoni and Oetzel, 1998). Typical diets fed to dry cows have a
DCAD of about +50 to +250 mEq/kg of diet dry matter (using equation 1). In common
feedstuffs, potassium is the most variable of the ions in the DCAD equation and it is
usually the most important determinant of DCAD in non-supplemented feed (Oetzel and
Goff, 1999). The successful use of dietary anions to prevent MF has suggested that diets
that are high in cations, especially Na and K, increase the susceptibility of cows to MF. A
good first step in formulating a low DCAD prepartum diet is to reduce dietary potassium
to less than 1.5% of dry matter. Once the cation content has been reduced as much as
possible by diet selection, anions can then be added to further reduce DCAD to the desired
end point (Oetzel and Goff, 1999). Commonly used anion sources are calcium chloride,
ammonium chloride, magnesium sulfate, ammonium sulfate, and calcium sulfate. Anionic
salts can be unpalatable and are always accompanied by a cation, which, depending on its
rate of absorption, will counteract some of the effects of the anions (Goff and Horst,
1998a, 1998b). Other anion sources include mineral acids such as hydrochloric or sulfuric
acid (Oetzel and Goff, 1999). However these acids in a liquid form are corrosive and
dangerous to handle. Commercial preparations of HC1 mixed into common feed


178
Guard, C. L. 1994. Cost of clinical disease in dairy cows. Page In Proc. Annual Cornell Conf.
Vet. Ithaca, NY. Cornell Univ., Ithaca, NY.
Gunnink, J. W. 1984. Retained placenta and leukocytic activity. Vet. Q. 6:49-51.
Gustaffson, A. H., L. Andersson, and U. Emanuelson. 1993. Effect of hyperketonemia,
feeding frequency and intake of concentrate and energy on milk yield in dairy cows Anim
Prod. 56:51-60.
Gustaffson, A. H., and U. Emanuelson. 1996. Milk acetone concentration as an indicator of
hyperketonemia in dairy cows: the critical value revised. Anim. Sci. 63:183-188.
Hall, M.B. 1997. Interpreting (and misinterpreting) feed analyses. Compend. Contin. Educ.
Prac. Vet. 19:S157-S161.
Hall, M.B., B.A. Lewis, P.J. Van Soest, and L.E. Chase. 1997. A simple method for
estimation of neutral detergent-soluble fibre. J. Sci. Food Agrie. 74:441-449.
Hall, M.B., A.N. Pell, and L.E. Chase. 1998. Characteristics of neutral detergent-soluble fiber
fermentation by mixed ruminal microbes. Animal Feed Science Technology 70:23-39.
Hayes, D.P., D.U. Pfeiffer, and N.B. Williamson. 1996. Effect of intraruminal monensin
capsules on reproductive performance and milk production of dairy cows fed pasture. J. Dairy
Sci. 79:1000-1008.
Hayirli, A., R. R. Grummer, E. Nordheim, P. Crump, D. K. Beede, M. J. VandeHaar, L. H.
Kilmer,, J. K. Drackley, D. J. Carroll, G. A. Varga, and S. S. Donkin. 1999. Prediction
equations for dry matter intake of transition cows fed diets that vary in nutrient composition.
J. Dairy Sci. 82(Suppl. 1): 113.
Hayirli, A., R. R. Grummer, E. V. Nordheim and P. M. Crump. 2002. Animal and Dietary
Factors Affecting Feed Intake During the Prefresh Transition Period in Holsteins. J. Dairy Sci.
85:3430-3443.
Heinrichs, A. J., D. R. Buckmaster, and B. P. Lammers. 1999. Processing, mixing, and
particle size reduction of forages for dairy cattle. J. Anim. Sci. 77:180-186.
Herbein J. H., R. J. Aiello, and E. L. Eckler. 1985. Glucagon, growth hormone, and glucose
concentrations in blood plasma of lactating dairy cows. J. Dairy Sci. 68: 320-325.
Herdt, T. H. 1988. Fuel homeostasis in the ruminant. Vet. Clin. North Am. Food Anim. Pract.
4:213-231.
Herdt, T. H. 2000. Ruminant adaptation to negative energy balance. Influences on the etiology
of ketosis and fatty liver. Vet. Clin. North Am. Food Anim. Pract. 16:215-230.


CHAPTER 4
EFFECT OF A MONENSIN-CONTROLLED RELEASE CAPSULE ON RUMEN AND
BLOOD METABOLITES IN FLORIDA HOLSTEIN TRANSITION COWS
Introduction
Monensin is an ionophore that alters the rumen microflora resulting in an increase in
molar proportion of propionic acid with a concurrent decrease in the molar proportion of
acetate and butyrate in the rumen (Richardson et al., 1976; Van Maanen et ah, 1978). The
increase in rumen propionate is accompanied by a reduction in the amount of methane
produced in the rumen and a lower incidence of ketosis and related disorders (Nagaraja et ah,
1997; Duffield et ah, 1998b; 1999b). Monensin also decreases L-lactate concentrations
(Callaway and Martin, 1997; Nagaraja et ah, 1997) and affects nitrogen metabolism by
decreasing rumen ammonia production in cattle fed different diets (Yang and Russell, 1993;
Nagaraja et ah, 1997).
Monensin has been available for use in food animals for over 20 years; however,
limited information is available about their effects in lactating dairy cows. During the last 5
years several studies have been conducted in Canada and Australia with the use of monensin as
a controlled release capsule in transition dairy cows (Stephenson et ah, 1997; Duffield et ah,
1998a, Duffield et ah, 1999a; Green et ah, 1999; Mutsvangwa et ah, 2002). Monensin is not
allowed in lactating dairy cattle in the United States.
Citrus pulp is an energy concentrate by-product rich in pectin and is a common
component of dairy cattle diets in Florida and other southern states (Hall, 1997; Arthington et
ah, 2002). Since pectin-fermenting bacteria are gram-negative monensin-resistant bacteria
118


36
The importance of bunk management practices that limit feed intake in the
etiology of LDA is likely to be greatest during the early postpartum period because of the
coinciding events of the transition period. TMR mixing can alter the physical form of the
fiber in the diet. Excess TMR mixing may grind coarse particles and cause a lack of fiber
physical form. Furthermore, excess particle size can allow the cows to sort the TMR in the
feed bunk which can also cause the same problem (Shaver, 1997; Vargas et al., 1998;
Heinrichs et al., 1999; Melendez et al., 2002; Melendez et al., 2003d).
Right dilatation of the abomasum and abomasal volvulus RDA-AV!
Right displacement and volvulus are less common conditions constituting 5 to 15%
of the total cases of displacements or volvulus (Trent, 1990).
Cattle with AV are more likely to appear systemically affected than those with
RDA or LDA. Shock, hypovolemia and pain may be present associated with distention
and necrosis of the abomasum, or severe electrolyte and acid-base imbalances, or both.
There is an abrupt decline in milk production. Feces are scant and often dark and
diarrhreal (Fecteau et al., 1999).
Many of the predisposing factors suggested for LDA, such as those that act by
altering motility or promoting gas build up, have been suggested as causative for RDA as
well. Right displacement of the abomasum is more diffusely distributed throughout the
lactation period than are LDAs (Trent, 1990). Both RDA and AV typically result in
hypochloremic, hypokalemic, metabolic alkalosis, paradoxical aciduria and are frequently
associated with hypocalcemia. However these metabolic changes are more pronounced in
AV than RDA (Fecteau et al., 1999).


58
Figure 2-1. Pathways of glucose, amino acids and fatty acids metabolism intersect at the
citric acid cycle
C3 Propylene
Glycol
Lactate
Glycerol
Alanine
Other Amino
Acids
C6 Glucose
Triglyceride
i
1
i
f
C16-C18 NEFAs
i
y
i
f
C2 Acetate
Aspartate
and other
aminoacids
Glutamate and other
amino acids
Adapeted from: Herdt, T. H. 1988. Fuel homeostasis in the ruminant. Vet. Clin. North
Am. Food Anim. Pract. 4:213-231.


64
rumen. Whereas bacteria flow completely from the rumen, protozoa flow only 20-40% of
that rate; however, approximately 25% of the microbial protein available to the host is
protozoal in origin. Approximately 1/4 to 1/3 of fiber degradation in the rumen is
protozoal. Bacterial ingestion and proteolysis by the protozoa decreases the efficiency of
nitrogen utilization by the host. An important consequence of the protozoa is their ability
to ingest starch and soluble sugars, preventing the alternative rapid bacterial fermentation
to lactic acid. They have an important role in regulating rumen lactate metabolism and in
preventing lactic acidosis. Some protozoa have been demonstrated to be sensitive to
monensin.
According to Van Soest (1994) and Orpin and Joblin (1997), yeast and anaerobic
fungi have long been known to be normal inhabitants of the rumen. Fungi are much more
particle-associated and involved in fibrolytic digestion than are protozoa. Fungi secrete a
more soluble cellulase complex than do rumen bacteria, and the mechanism of enzyme-
substrate interaction differ as well. The fungi produce volatile fatty acids, gas, and traces
of ethanol and lactate.
Volatile Fatty Acids
Volatile fatty acids (VFA) are end products of anaerobic microbial metabolism in
the rumen (Nagaraja et al., 1997). Major VFA in descending order of abundance are
acetic, propionic, butyric, isobutyric, valeric and isovaleric. The proportion of acetic,
propionic, and butyric acids can be markedly influenced by diet and the status of the
methanogen population in the rumen. Lactic acid is important when starch is part of the
diet, and is itself fermented to acetate, propionate, and butyrate (Van Soest, 1994). Rumen
concentrations of VFA are regulated by a balance between production and absorption
whereby increased production rate induces higher VFA concentrations (Nagaraja et ah,


145
cows. In addition, BCS did not change significantly from assignment to calving in treated and
control primiparous cows, respectively. Overall BCS at assignment was 3.41 0.02 and 3.45 0.02
between controls and treated primiparous cows respectively; while it was 3.40 0.02 and 3.45
0.02 for control and treated primiparous cows at calving, respectively. Treatment was significant for
BCS at calving in multiparous cows (P < 0.05). Overall BCS at assignment was 3.10 0.02 and
3.12 + 0.02 for control and treated multiparous cows, respectively; while it was 3.08 0.02 and 3.35
0.02 for control and treated multiparous cows at calving, respectively. The lack of effect of
monensin in primiparous cows might be explained because pregnant heifers are not mature animals
and they are still growing. In dairy heifers, monensin neither affects daily weight gain nor change
BCS or body composition (Meinert et al., 1992). Conversely, in the present trial, monensin did
affect BCS at calving in adult cows (multiparous); therefore monensin might have altered the tissue
composition of the daily gain in adult cows favoring the deposition of fat instead of protein. Gains
by more mature cattle contain more fat, less protein, and less water than gains by less mature cattle
(Williams et al., 1989). Indeed, in adult cows monensin increased either body weight, BCS or
reduced BCS losses after calving (Duffield et al., 1998a; Van Der Werf et al., 1998; Phipps et al.,
2000).
Milk Yield
In Table 5.3 least square means and SEM for accumulated current lactation and ME 305
milk yield for control and treatment groups within parity are shown. Illustrated in Figures 5.2 and
5.3 are test day milk yield for treated and control primiparous and multiparous cows, respectively.
There were no differences in test day, accumulated and ME 305 milk yield between groups
within parity (P > 0.05). These results are consistent with other studies in which monensin did not
improve milk production (Abe et al., 1994; Green et al., 1999; Vallimont et al., 2001).


68
Citrus Pub and Pectin Fermentation
Citrus pulp is an energy concentrate by-product produced in subtropical regions,
of which south central Florida remains the largest area of production (Arthington et al.,
2002). Citrus pulp is composed of pectin, that is galacturonic acid, arabinose, galactose,
and rhamnose, which are not digested by mammalian enzymes, but can be rapidly
fermented by ruminal microbes (Hall, 1997).
Pectin can inflate ADF values when rich-pectin feeds, like citrus pulp, are
analyzed by using traditional laboratory methods. Plant carbohydrates can be divided into
the neutral detergent fiber fraction (NDF) and the neutral detergent soluble carbohydrate
fraction (NDSC). The NDF fraction is composed of hemicellulose and cellulose. The
NDSC fraction is composed by organic acids, sugars, starches, fructans, pectin and B-
glucans. Fructans, pectin and B-glucans form the neutral detergent soluble fiber fraction
(NDSF). This fraction can be determined by a simple method that gives good precision
(Hall et al., 1997; Hall et al., 1998). Citrus pulp contains about 34.5 % of NDSF, while
forages contain between 12 to 22% (Hall et al.,.1997).
From in vitro fermentation experiments citrus pulp gives the lowest acetate to
propionate ratio and the highest propionate molar proportion as compared with beet pulp,
soybean hulls, mature and immature alfalfa stems, and mature and immature alfalfa leaves
(Hall et al., 1998).
Diets based on citrus pulp consistently demonstrate an increase in milk fat
content and milk urea nitrogen, but not in milk yield, when compared with diets richer in
starch (Belibasakis and Tsirgogianni, 1996; Leiva et al., 2000). Although higher levels of
milk fat have been suggested to be related to a higher production of acetate in the rumen,
molar proportion and concentration of acetate and propionate did not differ between
experimental diets containing citrus pulp and com, respectively (Leiva et al., 2000).


Risk of non-pregnancy
117
Days postpartum
Figure 3.8.Survival curves for risk of non pregnancy by treatment


41
Reasons for culling are death, selling for dairy purposes, low production, poor
reproduction, udder problems, feet and legs problems and miscellaneous (Radostits et al.,
1994). Death was associated with downer cow syndrome. Low production was explained
by low PTA for milk, previous milk production per day and calving season (summer);
poor reproduction was explained mainly by number of services, calf survival, cystic
ovaries and abortion; udder problems were explained by previous milk per day, current
milk per day, mastitis and teat problems; feet and legs problems were mainly explained by
PTA Milk, dystocia and foot or leg problem; miscellaneous reasons were explained by
PTA milk, LDA and mastitis (Milian-Suazo et al., 1989).
In New York Holstein state, older Holstein cows were at much higher risk of
being culled than younger animals. Calving season had no effect on culling. Higher milk
yield was protective against culling. Once a cow had conceived again, her risk of culling
dropped sharply. In all models, mastitis was an important risk factor throughout lactation.
Milk fever, RFM, displacement of abomasum, ketosis and ovarian cysts also significantly
affected culling at different stages of lactation. Metritis had no effect on culling. These
results indicated that diseases have an important impact on the actual decision to cull and
the timing of culling. Parity, milk yield, and conception status are also important factors in
culling decisions (Grdhn et al., 1998). In Finnish Ayrshire cattle, the farmers knowledge
of the cows pregnancy status had a significant effect on culling. The earlier the farmer
knew a cow was pregnant, the smaller was the risk of culling. If a cow had not been bred
at all, her risk of culling was 10 times higher than if she was inseminated once. The effect
of parity decreased when pregnancy status and number of inseminations were added to the
model. Mastitis, teat injuries and lameness had the greatest effect on culling, followed by
anestrus, ovarian cysts and MF. In general, the effect of diseases decreased when


97
Where:
7i = log of the odds of the event (e.g. metritis yes, no)
a = intercept
fh = parameter of Xi
Xi = treatment effect
p2 = parameter of X2
X2 = parity effect
pk = parameter of Xk
Xk = any explanatory variable, continuous or dummy
Results
In Table 3.3 incidence of calving-related disorders are reported. In Table 3.4 a
summary of logistic regression modeling is shown. Only dystocia and metritis were
affected by treatment. Cows treated with monensin were 2.1 times more likely to
develop dystocia than control cows (p < 0.01). First parity cows were 0.65 times less
likely to experience dystocia than older cows.
Correcting for parity, and dystocia, treated cows were 0.2 times less likely to
develop metritis than control cows (p < 0.01). Cows experiencing dystocia, or RFM or
cows in first lactation were 20.2, 27.3 or 5.7 times more likely to develop puerperal
metritis than cows calving unassisted, without RFM or older cows.
Figure 3.1, illustrates BCS of treated and control cows at assignment and at calving.
Body condition score at assignment and previous lactation milk production were similar
between groups within parity (P > 0.05). There was an interaction between treatment
and parity. Monensin did not increase BCS at calving in first lactation cows (P > 0.05),
but did increase it in older cows (P < 0.01).
In Figures 3.2 and 3.3 daily milk yield up to 20 d pp by parity and treatment are
shown. Within parity 1, milk yield was not different between groups (P > 0.05). Within
parity 2, treated cows produced more milk than control cows at day 3 (P < 0.05).


104
might be assumed to be real. Mechanism for this positive effect might be related to the
improvement of energy balance mediated by monensin, through a better BCS at calving,
and attributable to the lower incidence of puerperal metritis found in treated cows.
It is concluded that a monensin controlled-release capsule applied at dry-off in
transition cows fed diets containing citrus pulp improved slightly milk yield, decreased
the incidence of metritis and increased the incidence of dystocia in third parity or older
cows. Milk fat was decreased by treatment. There was a tendency for a higher
conception rate at first service in cows treated with monensin, independent of parity.


80
Increasing dietary NFC or decreasing NDF during the transition period stimulates
DMI. When energy density of the diet increased from 1.3 to 1.54 Meal ENl/kg DM and
crude protein increased from 13 to 16% at about 3 weeks prior to calving, DMI increased
in 30% (Emery, 1993). Plasma fatty acids decreased from 346 to 228 pM and liver
triglyceride decreased from 15 to 9 mg/g wet tissue (Dyk et al., 1995; VandeHaar et ah,
1995). In another study, cows fed high NFC (40-42%) diets consumed more dry matter
during the prepartum period, had lower plasma NEFA, had reduced BHB level and there
was a reduction in liver triglyceride (Minor et ah, 1996). Finally, VandeHaar et ah (1999)
found that increasing nutrient-density of prepartum diets did not decrease DMI. There
were less NEFA in plasma (176 ws. 233 pM) and more insulin-like growth factor-I (472
vs. 390 ng/ml plasma) during the last two weeks prepartum and less triglyceride in liver at
parturition (0.9 vs. 1.5% wet tissue basis). They concluded that increasing the energy and
protein density up to 1.6 Meal of NEl/kg and 16% CP in diets during the last month before
parturition improves nutrient balance of cattle prepartum and decreases hepatic lipid
content at parturition.
Protein
The protein concentration needed in the diet to meet requirements for maintenance
and gestation during the transition period is shown in Figure 2-3.
The results about crude protein levels in precalving cows are controversial. Curtis
et ah (1985) indicated that feeding protein above NRC (1989) during the final 3 weeks
prepartum decreased the risk of RFM and uncomplicated ketosis. Mahanna (1997)
recommends increasing CP to 14.5% 15.5% to begin preparing for high DMI during the
lactation phase. However, Santos et ah (1999a,b) found precalving heifers fed CP up to
14.7% benefited with higher milk yield and milk fat production, but precalving cows did


128
Table 4-2. Nutrient content of dry cow far-off, dry-cow transition and lactating
transition diets
Nutrient
Dry cow
far-off
Dry cow
transition
Lactating
transition
CP(%DM) '
15.52
17.83
18.70
UndegP(%CP)2
26.38
34.35
31.34
DegP(%CP) 2
73.62
65.65
68.66
SolP(%CP)2
-
35.72
38.77
NEL(Mcal/kg)3
0.84
1.67
1.69
ADF(%DM) '
24.54
24.13
23.70
NDF(%DM)1
36.04
35.20
34.53
NFC(%DM) 2
32.94
32.35
34.56
Starch(%DM)2
11.05
13.99
15.10
Lipid (%DM) 1
2.85
5.89
4.36
Ca(%DM)1
0.74
1.29
1.15
P(%DM) 1
0.30
0.36
0.45
Mg(%DM)1
0.32
0.36
0.37
K(%DM) 1
1.28
1.12
1.48
Na(%DM) 1
0.09
0.09
0.56
C1(%DM)1
0.18
0.45
0.45
S(%DM) 1
0.24
0.41
0.21
Forage in diet (%DM)
61.49
55.72
48.94
Cation-Anion (meq/kg DM)4
165.90
-58.50
363.10
1 Laboratory nutritional analysis
2 Values from feed composition tables
3 From formulas after laboratory analysis
4From formula (Na+ + K+) (CT + S')


71
In another Australian study, where the controlled-release capsule of monensin
was given at 50 7 days before expected parturition, Stephenson et al. (1997) found that
treated cows (n=12) had lower plasma concentrations of glucose, NEFA and BHB than
control cows (n=12) before calving. However, no significant differences in plasma
concentrations of glucose, NEFA and BHB were found between groups after parturition.
Cows were fed on pasture and supplemented concentrate at a level of 4 to 10 kg/cow/day.
Later, Beckett et al. (1998) conducted a randomized clinical trial including 1109
cows from 12 Australian dairy herds. They postulated that treatment administered as an
intraruminal bolus 40 days before expected parturition and 50 days after calving would
reduce the risk of postpartum reproductive failure, reduce the incidence of common
periparturient clinical disorders, and increase the production of milk and milk solids.
Treatment did not significantly alter any reproductive outcome; 54.5% of treated cows and
58.2% of control cows were pregnant at first service. Treatment with monensin did not
reduce the risk of dystocia, ketosis, milk fever, infectious diseases, metritis, RFM,
lameness, or abortion nor did treatment increase the risk of mastitis. However, milk
production was increased in treated cows (0.75 L/d) after adjustment for herd. Milk fat
yield was similar between groups.
In New Zealand, cows from 3 herds fed on pasture received a controlled-release
capsule of monensin 1 month prior to the start of artificial insemination. The objectives of
the study were to determine whether treatment with controlled-release capsules of
monensin improved milk production and reproductive performance of cows calving in
spring and fed pasture. Treated cows produced more fat, protein and milk per day (19.1 vs
17.7 L) during the second month of monensin activity than control cows. Overall
pregnancy rate to first and second service were not different between treated and control


11
Table 2-3 Summary of relationships among calving-related disorders
Author
Disease
Risk factors
Association
Curtis et
Milk fever
Parity
Positive
al., 1984
Estimated transmited
Positive
ability
Increased ccrude protein
in dry period
Negative
Erb et al.,
RFM
Milk fever
OR= 2.0
1985
Parity
Positive
Milk fever
Parity
Positive
Metritis
Milk fever
OR= 1.6
REM
OR= 5.8
Curtis et
Milk fever
Parity
Positive
al., 1985
RFM
Milk fever
p
'i
il
Pi
O
Parity
Positive
Metritis
RFM
OR= 5.7
LDA
OR= 3.6
LDA
Ketosis
OR= 11.9
Ketosis
LDA
OR= 53.5
REM
OR= 16.4
Milk fever
OR= 23.6
Grohn et
Milk fever
Parity
Positive
al., 1989
Milk yield
Positive
Udder edema
Parity
Negative
Milk yield
Positive
RFM
OR= 2.6
Mastitis
OR= 3.8
Milk Fever
OR= 2.5
Abomasum
Ketosis
OR= 5.7
disorders
Hypomagnesemia
OR= 7.0
RFM
OR= 2.4
Metritis
OR= 2.5
Mastitis
OR= 3.6


113
DHIA Test Day
Figure 3-4. Least squares means and SEM for test day milk yield by parity 1 and 2


187
Rukkwamsuk, T., T.A.M. Kruip, G.A.L. Meijer and T. Wensing. 1999c. Hepatic fatty acid
composition in periparturient dairy cows with fatty liver induced by intake of a high energy
diet in the dry period. J. Dairy Sci. 82:280-287.
Rukkwamsuk, T., T. Wensing, and M. J. H. Geelen. 1998. Effect of overfeeding during the
dry period on regulation of adipose tissue metabolism in dairy cows during the periparturient
period. J. Dairy Sci. 81:2904-2911.
Rukkwamsuk, T., T. Wensing, and M. J. H. Geelen. 1999a. Effect of overfeeding during the
dry period on the rate of esterification in adipose tissue of dairy cows during the periparturient
period. J. Dairy Sci. 82:1164-1169.
Rukkwamsuk, T., T. Wensing, and M. J. H. Geelen. 1999b. Effect of fatty liver on hepatic
gluconeogenesis in periparturient dairy cows. J. Dairy Sci. 82:500-505.
Russel, J.B. and S.A. Martin. 1984. Effect of various methane inhibitors on the fermentation
of amino acids by mixed rumen microorganisms in vitro. J. Anim. Sci. 59:1329-1338.
Santos, J. E. P., P. W. Jardon, E. J. DePeters, and J. T. Huber. 1999a. Effect of prepartum
crude protein level on performance of primiparous Holstein cows. Page 200. In Proceedings
32th Annual Convention America Association of Bovine Practitioners. Nashville, TN.
Santos, J. E. P., P. W. Jardon, E. J. DePeters, and J. T. Huber. 1999b. Effect of prepartum
crude protein level on performance of multiparous Holstein cows. Page 201. In Proceedings
32th Annual Convention America Association of Bovine Practitioners. Nashville, TN.
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Sauer, F. D., J. K. G. Kramer, and W. J. Cantwell. 1989. Antiketogenic effects of monensin
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Schelling, G.T. 1984. Monensin mode of action in the rumen. J. Anim. Sci. 58:1518-1527.
Seal, C.J., and C.K. Reynolds. 1993. Nutritional implications of gastrointestinal and liver
metabolism in ruminants. Nutr. Res. Rev. 6:185-208.
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3
monensin does not affect pectin-fermenting bacteria, it is reasonable to assume that dairy
cows fed monensin and diets rich in citrus pulp should not modify extensively their
pectin rumen fermentation. In addition, monensin still would improve starch and other
soluble carbohydrate rumen fermentation, with the consequent increase in rumen
propionate and blood glucose and decrease in ketone body formation and fat
mobilization.
Therefore, the objectives of this research program were
To determine the effect of a monensin controlled-release capsule inserted at dry-off
on the incidence of calving-related disorders, milk production and reproductive
responses in transition dairy cows fed diets containing citrus pulp.
To establish the effect of a monensin controlled-release capsule inserted at dry-off on
rumen metabolites and blood energy-related metabolites in transition dairy cows fed
diets containing citrus pulp.
To evaluate a milk ketone body test strip and to determine the incidence of subclinical
ketosis at 14 days postpartum in transition dairy cows fed diets containing citrus pulp
and that received a bolus of monensin at dry-off.


157
days BEP. Far-off cows were fed a typical Florida dry cow diet (Table 6.1 and 6.2). Close-up
dry cows (21 days BEP) were housed in a dry-lot with adequate feed bunk space and shade.
They received twice a day, a diet containing citrus pulp with a DC AD of-51.7 mEq/kg DM
using the equation DCAD (mEq)= (Na + K) ( Cl + S) (Tables 6.1 and 6.2).
Cows were processed within 12 h postpartum, on a routine basis, which included
recording BCS, udder score (for presence of edema), reproductive tract status (trauma or
lacerations), and whether the cow was suspected of having retained fetal membranes. If cows
developed either RFM or milk fever they were treated and remained in the hospital bam until
recovery. After calving processing, cows were moved to a postpartum lot and fed a diet higher
in forage NDF (Tables 6.1 and 6.2). Any cow with decreasing milk yield was moved from the
milking herd to the hospital bam to be examined and treated as needed.
Beginning 60 days postpartum, cows received a bST protocol (Posilac, 500 mg
sometribove zinc, subcutaneously; Monsanto, St Louis, Missouri) every 14 days during the
entire lactation.
Experimental Protocol
During July to August 2001, 300 cows dried-off 50 to 70 days BEP were randomly
assigned either to treatment or control group. The treated group (n=150) received orally a
capsule of monensin (releasing 300 mg of monesin daily for 95 days, CRC Rumensin ,
ELANCO Animal Health, Guelph, ON, Canada). Control cows (no capsule, n=150) were
randomly matched by parity.
The number of animals per treatment was calculated expecting a reduction in the proportion
of cows with concentrations of BHB in milk > 200 pmol/L at 14 days postpartum from 25% to
18% (95% confidence, 80% of power).


78
diseases and the most effective ways to group and manage dry cows. Critical physiologic
events that have to be targeted during the transition period include adaptation of the rumen
to the higher energy diet that will be fed in early lactation, maintenance of normal blood
calcium concentration, a strong immune system throughout the peripartum period, and
maintenance of a slightly positive energy balance up to the time of calving (Goff and
Horst, 1997b; Oetzel, 1998).
Energy
Energy balance of a transition cow is determined by subtracting energy
requirements for maintenance and gestation from energy intake. During the transition
period, feed intake is decreasing at a time when energy requirements are increasing due to
growth of the conceptus. Consequently, to maintain the energy balance the energy density
of the diet should increase (Grummer, 1999). In Figure 2-2 the net energy of lactation
(NE1) needed in diets of cows and heifers to meet requirements for maintenance and
gestation during the transition period is shown. Heifers need higher dietary energy density
due to lower feed intake and the additional energy requirements to support growth (NRC,
2001).
Increasing energy density may stimulate papillae growth and increase acid
absorption from the rumen, adapt the microbial population to higher starch diets, increase
blood insulin, decrease fatty acid mobilization from adipose tissue and increase dry matter
intake (Grummer, 1999).
Grain has to be introduced to the cows ration for at least 3 weeks before the due
date and for heifers this should be 5 weeks. The energy density should be between 1.56-
1.62 Mcal/kg of NE1 (NRC, 2001).


24
collagenization of connective tissue, reduction in blood supply, appearance of polynuclear
giant cells, loosening of tissues, and contraction of the uterine musculature (Grunert, 1986;
Youngquist and Braun, 1993). Fifty-nine percent of RFM were spontaneously expelled
between 5 and 7 days postpartum and 94% at days 11-13 postpartum (Van Werven et al.,
1992). Most RFM are expelled by 4 to 10 days postpartum after sufficient necrosis of the
caruncular tissue (Youngquist and Braun, 1993). Significant changes in the placentome
must take place before expulsion of the fetal membranes can occur. Increased collagenase
and other protease activities have been described in the normal uterus. This has resulted in
a massive breakdown of collagen and other proteins during uterine involution (Sharpe et
al., 1990). However, in cows experiencing RFM collagenolysis and proteolysis is
diminished (Eiler, 1997).
Some studies have reported an average incidence of RFM from 4 to 11% with a
range of 2-55% (Paisley et al., 1986; Joosten et al., 1988). The incidence of RFM, based
on 50 citations from 1979 to 1995 ranged from 1.3% to 39.2%. The median was 8.6%
(Kelton et al., 1998). Economic losses of a case of RFM, including direct cost such as
treatment (32%), lost milk production (40%), increased culling rate (19%) and increased
days open (9%), have ranged from $106 to $285 (Barlett et al., 1986a;Joosten et al., 1988;
Guard, 1994).
Joosten et al., (1991) have reported a number of risk factors for RFM such as
dystocia, parity, abnormal gestation duration, season and sire of the calf. In other studies,
cows that developed parturient paresis were 4.0 to 4.2 times more likely to experience
RFM than normal cows (Curtis et al., 1985; Erb et al., 1985).
Hypocalcemia has been related to dystocia, RFM and uterine prolapse in dairy
cattle (Curtis et al., 1983; Risco et al., 1984; Correa et al., 1993; Risco et al., 1994a). Erb


Milk protein (%)
116
Figure 3-7. Least squares means and SEM for test day protein % by parity 3 or
greater


98
The effect of treatment and parity on test day milk yields for the entire lactation
is presented in Figures 3.4 and 3.5. For first parity cows differences were not statistically
significant (P > 0.05). For older cows, monensin improved milk yields with significant
differences at test day 4 and 7 (P < 0.05). Accumulated milk yield of the entire lactation
were not different in cows treated with monensin than controls (13910 185 vs 13627
185, respectively) (P > 0.05). In Figures 3.6 and 3.7 test day milk fat % and milk protein
% for cows in lactation 3 or greater by treatment are illustrated. There is clear evidence
that milk fat% was lower in cows treated with monensin than controls. For percentage of
milk protein, control cows had similar levels than cows treated with monensin (P >
0.05).
Reproductive responses for treated and control cows are shown in Table 3.5 and
Figure 3.8.
Discussion
Calving-related disorders
The incidence of MF reported in this study was consistent with the use of anionic
salts in the prepartum period (0%) (Block, 1984; Oetzel et al., 1988; Beede et al., 1991;
Goff et al., 1991; Block, 1994; Vagnoni and Oetzel, 1998).
The incidence of RFM of about 20% reported in the present study was relatively
high, but consistent with a previous study conducted on the same farm (Melendez et al.,
2003b) and within the range of 1.3 to 39.2% cited in the literature (Kelton et al., 1998).
The lack of treatment effect was expected since RFM is a multifactorial condition and
other variables are more important predisposing factors for this CRD (Paisley et al.,
1986; Stevenson and Call, 1988; Correa et al., 1993; Eiler, 1997).


92
is the increased molar proportion of propionic acid with a concurrent decrease in the
molar proportion of acetate and butyrate in the rumen (Richardson et al., 1976). As a
result, a lower incidence of ketosis and glucose-lipid metabolism-related disorders has
been reported (Duffield et al., 1998b; 1999b).
Monensin has been available for use in food animals for over 20 years. However,
limited information is available about its effect in lactating dairy cows. During the last 5
years, several research studies have been conducted in Canada and Australia, on the use
of a monensin as a controlled release capsule (Stephenson et al., 1997; Duffield et al.,
1998a, Duffield et al., 1999a; Green et al., 1999; Mutsvangwa et al., 2002). However,
monensin is not approved for use in lactating dairy cattle in the United States.
Since pectin-fermenting bacteria are gram-negative monensin-resistant bacteria
(Nagaraja et al., 1997; Stewart et al., 1997), it is hypothesized that Holstein dairy cows
supplemented with monensin and fed diets containing citrus pulp will have a decrease in
the incidence of calving-related disorders, and an increase in milk production.
In order to determine the effect of a monensin-controlled release capsule
administered at dry-off on the incidence of calving-related disorders, fertility and milk
yield in Holstein transition dairy cows fed citrus pulp-based diets the following field
trial was conducted.
Materials and Methods
Cows and herd management
The study was conducted on a commercial dairy farm with 3600 milking cows
located in north central Florida, with a milk rolling herd average of 10,700 kg. Most
lactating cows were housed in a dry-lot system and were fed the same TMR three times
a day, except postpartum transition cows which received a diet higher in forage NDF.


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Bolinder, A., B. Seguin, H. Kindhal. 1988. Retained fetal membranes in cows: manual
removal versus nonremoval and its effect on reproductive performance. Theriogenology
30:45-56.


39
lameness was calculated at 0.16. The genetic and phenotypic correlation between 305-d
milk yield and lameness was 0.24 and 0.04 respectively (Van Dorp et al., 1998).
Prevalence and incidence of lameness have been reported worldwide with a large
variation in the values. Different methodologies have been used and results must be
interpreted with caution. The frequency of lameness, based on 39 citations from 1972 to
1995 ranged from 1.8% to a mean annual incidence rate of 30% (Kelton et al., 1998).
Wells et al. (1993), in Wisconsin and Minnesota, Kaneene and Hurd (1990), in Michigan,
and Miller and Dorn (1990), in Ohio reported an annual incidence of lameness of 6.6, 6.9,
and 5.1 cases per 100 cows at risk, respectively. Throughout the United States, the USDA-
APHIS (1997) showed that the annual incidence of lameness in herds less than 100 cows
was 15.2% and over 100 cows was 18.8%. The region with the highest incidence was the
Northeast (21.2%) and with the lowest incidence was the Southeast (8.6%). In Florida, at
the Dairy Research herd of the University of Florida, the incidence of lameness was
determined to be 51 cases per 100 cows in a year (Shearer and Elliot, 1998).
The cost of lameness is related to treatment cost, losses in milk production,
discarded milk, decrease in dry matter intake, reproductive problems, culling, opportunity
costs and susceptibility to other diseases. A cost of a case of interdigital lameness has been
calculated as $160, a case of digital lameness $392 and a case of sole ulcer $700
(Greenough et al., 1997). Guard (1994) estimated an average cost of a case of lameness to
total approximately $302. Deluyker et al. (1991) reported that lame Holstein cows had a
drop in milk production of 1.7 kg/day during a 4-week period around diagnosis. In Finish
Ayrshire cows, Rajala-Schultz et al. (1999b) found that lameness affected milk
production. In this study, cows with foot and leg disorders experienced milk losses that


96
measures. Body condition score at calving was analyzed by ANOVA mixed model.
Overall pregnancy rate was analyzed by survival analysis. Survival functions and curves
were developed for each group. Conception rate at first service was analyzed by logistic
regression as described for CRD. Calving to conception interval and services per
conception were analyzed by ANOVA mixed model. In both logistic regression models
and mixed models variables were considered significant at P < 0.05. Statistical analysis
was conducted using SAS 8.2 (SAS, 2001).
ANOVA, Mixed models:
yijk= p + T¡ + Park + (Par*T)kj + eijk
Where:
y¡jk= body condition score, actual and ME305 milk yield
Ti= fixed effect of treatment
Cow(Tj)j = random effect of cow nested in treatment
Park = fixed effect of parity
(Par*T)k¡ = fixed effect of interaction parity and treatment
e¡jk random error term
Mixed model for repeated measures:
YijkinW P + T¡ + Pap + Timek + BCSm + (Par*Time*T)kn + ejjk|m
Where:
YijkinW milk yield
Tj= fixed effect of treatment
Cow(Tj)j = random effect of cow nested in treatment
BCSm = fixed effect of body condition score
Timek = fixed effect of time
(Par*Time*T)¡ki= fixed effect of triple interaction parity, time and treatment
6¡jkim = random error term
Logistic regression model:
Logit (n) = a + PiX] + P2X2 + .... pkXk


32
depressed in early lactation compared with the curves for cows without LDA. Cows with
LDA experienced severe losses of milk yield because the disease occurred during peak
yield, which might explain the severity of the losses. Milk yield returned to a normal range
at 20 to 45 d after diagnosis. Furthermore, cows with LDA were 1.8 times more likely to
have another disease than healthy cows. 0stergaard and Grdhn (1999), in Denmark, found
that cows with LDA compared with healthy cows had an average milk loss within the first
6 weeks after diagnosis of 4.6 and 5.2 kg/d for primiparous and multiparous cows,
respectively. Three weeks before diagnosis, only multiparous cows with LDA showed
lower milk yield than normal cows (approximately 4.0 kg/d). Also, this study
demonstrated cows that experienced LDA presented the highest body weight loss (69 kg)
and the pre-disease level for accurate estimation of losses has to be considered.
Displaced abomasum is also a risk factor for culling in early lactation (1 to 30
days), but not in late lactation. Cows with LDA had a relative risk of culling 2.4 times
higher than normal cows during the first 30 days of lactation. After that period of lactation
the differences were not significant (Grhn et al.,1998).
Left displacement of the abomasum is a multifactorial disease where different risk
factors have been established. Several epidemiological and clinical trials have been
conducted to determine association among factors and physiopathological mechanism.
Cows experiencing MF, dystocia and ketosis increased the odds for LDA by 2.3, 2.3 and
13.8 respectively, when compared to normal cows (Correa et al., 1993). In another study
conducted by Cameron et al., (1998) in Michigan, individual cow and herd risk factors
were examined. Significant factors associated with an increased risk of LDA in the
individual cow model included high body condition score, winter season, and plasma
NEFA concentration > 0.3 meq/L between 35 and 3 d prepartum. The risk of LDA


65
1997). Volatile fatty acid production rates vary diumally as a consequence of eating
pattern, therefore VFA rumen concentrations and pH vary as well. Fermentation peaks
about 4 hours after feeding on a hay diet but occurs sooner if the diet contains some
concentrate. The major factor affecting VFA absorption is their concentration (Van Soest,
1994). In general, acetate concentration in the rumen ranges between 60 to 70 mmol/L,
propionate between 18 to 25 mmol/L and butyrate between 10 to 15 mmol/L (Nagaraja et
al., 1997)
Monensin and Rumen Fermentation
The most consistent and well-documented fermentation alteration observed upon
monensin feeding is the increased molar proportion of propionic acid with a concurrent
decrease in the molar proportion of acetate and butyrate in the VFA produced in the rumen
(Richardson et al., 1976). Addition of cellobiose and monensin to an in vitro fermentation
batch demonstrated that total concentrations of volatile fatty acids were increased.
Monensin treatment increased propionate and total VFA concentrations and decreased L-
lactate concentrations. A lag in organic acid utilization was observed in fermentations,
which was most likely due to bacteria that were resistant to monensin and that
preferentially utilized soluble nutrients other than organic acids (Callaway and Martin,
1997).
The increase in rumen propionate is accompanied by a reduction in the amount of
methane produced in the rumen (Nagaraja et al., 1997). Ionophore antibiotics are not
inhibitors of methanogenic bacteria. They are believed to reduce precursors of
methanogenesis (H2) Co2 and formate) (Russel and Martin, 1984); however monensin
inhibits methanogenesis from formate, probably as a result of inhibition of nickel uptake


88
Table 2-9. Minimum requirements for dry, and prepartum cows
Nutrient
Far-off dry cows
Close-up dry cows
Dry matter intake (kg/day)
14.4
13.7
Net energy for lactation
1.25
1.54-1.62
(Mcal/day)
Maximum crude fat (%)
5
6
Crude protein (%)
13
14-15
Undegradable intake
25
32
protein (% CP)
Acid detergent fiber (%)
21
17-21
Neutral detergent fiber (%)
33
25-33
Minimum forage NDF (%)
30
22
Maximum NFC (%)
36-43
36-43
Minimum Calcium (%)
0.44
0.45
Phosphorus (%)
0.22
0.3-0.4
Ca:P ratio
1.5:1 to 5:1
1.5:1 to 5:1
Magnesium (%)
0.20
0.35-0.4
Potassium (%)
0.55
0.55
Sulfur (%)
0.11
0.11
Sodium (%)
0.10
0.10
Chlorine (%)
0.20
0.20
DCAD (mEq/kg)
-
<0
^^^^i^^^
Adapted from: National Research Council. 2001. Nutrient requirements of dairy cattle, 7th
revised edition. Washington, D.C.: National Academy Press.


NEFA (mEq/L) Glucose (mmol / L) BHB (mmol/L)
154
Figure 5-5. Glucose, beta-hydroxy butyrate (BHB), and non-esterified fatty acids
(NEFA) concentrations over time in multiparous monensin treated and
control.


38
or environmental mastitis. The most common contagious mastitis pathogens are
Streptococcus, agalactiae and Staphylococcus, aureus. Environmental mastitis refers to
infections caused by two categories of organisms, coliform and streptococcal species
(Radostits et al., 1994). There is increasing evidence that, as the contagious pathogens are
progressively controlled in a herd, the incidence of clinical cases caused by coliforms
organisms increases (Jones, 1990).
Mastitis is a multifactorial disease where three major factors are involved: host
resistance, microbial agents, and the environment. The general resistance by the host is
related to genetic predisposition, anatomic characteristics, nutritional status, stage of
lactation and parity (Radostits et ah, 1994). Periparturient diseases, such as dystocia,
parturient paresis, RFM and ketosis have all been identified as risk factors for the
subsequent development of mastitis. Cows with MF are 5.0 times more likely to develop
clinical mastitis than normal cows (Curtis et ah, 1983).
Lameness
Lameness is one of the most prevalent disease in dairy cattle operations (Shearer,
1996) Lameness is a multifactorial condition and costly disease where many factors have
been identified. Nutrition, genetics or conformation, facilities, environment and hygiene,
behavior and management are the most important risk factors in the etiology of lameness
in cattle (Greenough et ah, 1997). As examples, in a study conducted at the University of
Florida, cows that were fed a diet low in prepartum energy and high in postpartum energy
developed subclinical laminitis. This feeding protocol was associated with a significantly
higher rate of rumen acidosis than rations with relatively low postpartum energy
(Donovan et ah, in press). In another study conducted in Canada, the heritability of


EFFECTS OF MONENSIN IN TRANSITION HOLSTEIN DAIR COWS FED DIETS
CONTAINING CITRUS PULP
By
PEDRO G. MELENDEZ
A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE
UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA
2004


I certify that I have read this study and that in my opinion it conforms to
acceptable standards of scholarly presentation and is fully adequate, in scope and
quality, as a dissertation for the degree of Doctor of Philosophy.
Jesse P. Goff
Researcher, USDA
This dissertation was submitted to the Graduate Faculty of the College of
Veterinary Medicine and to the Graduate School and was accepted as partial
fulfillment of the requirements for the degree of Doctor of Philosophy.
May, 2004
Dean, College of Veterinary
Medicine
Dean, Graduate Sthool


Body Condition Score
110
3.5
3.25 -
2.75 -
2.5
Prim. Con
Prim. Mon
A Mult. Con
- X- Mult. Mon
Dry-off
Calving
Figure 3-1. Body Condition Score by treatments and by parity at
assignment and at calving. ** Interaction parity by treatment.
BCS statistically different at calving in multiparous cows (P <
0.05)


74
During the last 5 years the group from University of Guelph, Canada, has
developed a strong line of research in transition dairy cows using a controlled-release
capsule of monensin starting in the prepartum transition period. The overall research
project used 1010 dry cows and pregnant heifers from 25 dairy farms near Guelph,
Ontario, Canada. The monensin capsule was inserted 3 weeks prior to expected calving
and released 300 mg of monensin during approximately 95 days. The first report
determined that monensin treatment significantly reduced serum BHB concentrations
between 150 and 200 pmol/L at week 1, 2 and 3 postpartum. Treatment also significantly
raised serum glucose concentrations during weeks 1 and 2 of lactation, with an increase in
serum glucose concentrations of 0.16 to 0.22 mmol/L. Monensin treatment significantly
reduced aspartate aminotransferase activity during the entire postcalving period as well,
especially in weeks 2 and 3 postpartum. In addition, monensin reduced the loss of BCS
after parturition. No significant treatment effects were found for calcium, phosphorus, or
total protein (Duffield et al., 1998a). The authors also reported that incidence and
prevalence of subclinical ketosis (BHB > 1200 pmol/L) were significantly reduced by
about 50% (odds ration 0.46) by monensin treatment. In addition, monensin was also
significant in reducing the incidence of subclinical ketosis by using two other serum
thresholds (1400 and 2000 pmol/L). The prevalence of subclinical ketosis was also lower
in the treated group than control cows when milk ketone tests were used as the diagnosis
method (Duffield et al., 1998b).
In a third report, the authors evaluated the effect of monensin on milk production
and milk components at the first three Dairy Herd Improvement Association (DHIA) tests.
Treatment with monensin increased milk production, but this effect was dependent on
body condition score prior to calving. Cows with a BCS <3.0 (scale 1-5) did not have


47
1999b). In addition, lipid accumulation dramatically decreases ureagenesis with increased
blood ammonia concentrations, which decreases gluconeogenesis from propionate but not
from alanine (Overton et al., 1999).
The entire process of ketogenesis, from adipose mobilization to mitochondrial fatty
acid transport, appears to be affected by the opposing actions of insulin and glucagon,
making the insulimglucagon (I:G) ratio an important determinant of ketogenesis. Cows in
early lactation have low serum insulin concentrations in the face of normal glucagon
concentrations, resulting in a reduction in the I:G ratio. This low I:G ratio promotes
adipose tissue mobilization and ketogenesis. Ketone bodies, however, appear to stimulate
insulin secretion and therefore may raise the I:G ratio, creating a negative feedback on
ketone body production (De Boer et al., 1985; Herbein et al., 1985; Herdt, 2000). The net
result of these changes and balances is that during periods of negative carbohydrate and
energy balances, ruminants normally have moderately elevated blood ketone body
concentrations and moderately depressed blood glucose concentrations (Herdt and Emery,
1992; Herdt, 2000).
In average in cows, dry matter intake decreases precipitously by 30% on day 1 or 2
before calving and does not recover until 1 to 2 days after calving. Liver triglycerides
(TG) are increased 6 fold by the day of calving (11.8% DM) as compared before calving.
By 4 weeks into lactation the liver TG are 5 fold higher than before calving (10.2% DM).
Plasma NEFA are 5 (1.1 mEq/L) and 1.5 (0.36 mEq/L) times prepartum levels (25 d
prepartum; 0.2 mEq/L) at calving and 35 days postpartum, respectively (Bremmer et al.,
2000). Triglyceride and lipid accumulation in the liver is a much earlier phenomenon than
previously assumed. Elevation of plasma NEFA concentrations starts prior to DMI
depression, on day 5 before parturition. Liver TG infiltration does not occur until the


107
Table 3-3. Incidence of calving-related disorders
Disorder
Lactational incidence (%)
Control
(n=290)
Monensin
(n=290)
Dystocia
17.9
30.9*
Milk fever
0.0
0.0
RFM 1
19.7
22.8
Metritis
42.6*
33.8*
Ketosis
2.5
2.8
DA2
1.4
0.7
Culling rate
28.6
28.6
Retained fetal membranes
2 Displacement of abomasum
*P < 0.05


180
Hocquette, J.F., and D. Bauchart. 1999. Intestinal absorption, blood transport and hepatic and
muscle metabolism of fatty acids in preruminant and ruminant animals. Reprod Nutr Dev
39:27-48.
Hoffman, P.C., N.M. Brehm, S.G. Price, A. Prill-Adams. 1996. Effect of accelerated
postpubertal growth and early calving on lactation performance of primiparous Holstein
heifers. J. Dairy Sci. 79:2024-2031.
Horst, R. L., J. P. Goff, T. A. Reinhardt. 1994. Calcium and vitamin D metabolism in the
dairy cow. J. Dairy Sci. 77:1936-1951.
Hosmer, D. W. and S. Lemeshow. 1989. Applied logistic regression. John Wiley & Sons, Inc.
New York.
Hutjens, M. F. 1991.Feed additives. Vet. Clin. North Am. Food Anim. Pract. 7(2): 525-540.
Hutjens, M. F. 1992. Selecting feed additives. Pages 309-317. In Large Dairy Herd
Management. H. H. Van Horn and C. J. Wilcox. American Dairy Science Association, 301
West Clark St., Champaign, IL 61820 .
Ingvartsen, K. L. and J. B. Andersen. 2000. Integration of metabolism and intake regulation: a
review focusing on periparturient animals. J. Dairy Sci. 83:1573-1597.
Johnson, M. M. and J. P. Peters. 1993. Technical note: An improved method to quantify
nonesterified fatty acids in bovine plasma. J. Anim. Sci. 71:753-756.
Jones, T. O. 1990. Escherichia coli mastitis in dairy cattle: a review of the literature. Vet.
Bull. 60:205-231.
Joosten I, Stelwagen J, Dojkhuizen AA. 1988. Economic and reproductive consequences of
retained placenta in dairy cattle. Vet. Rec. 123:53
Joosten L, P. van Eldik, and G. J. W. van der Mey. 1991. Factors affecting retained placenta
in cattle. Effect of sire on incedence. Anim. Repro. Sci. 25:11-22.
Jorgensen, N. A. 1974. Combating milk fever. J. Dairy Sci. 57:933-944.
Jorritsma, R., S. J. Baldee, Y. H. Schukken, T. Wensing, and G. H. Wentink. 1998. Evaluation
of a milk test for detection of subclinical ketosis. Vet. Q. 20:108-110.
Joyce, P. W., W. K. Sanchez, and J. P. Goff. 1997. Effect of anionic salts in prepartum diets
based on alfalfa. J. Dairy Sci. 80:2866-2875.
Kaneene, J. and H. Hurd. 1990. The National Animal Health Monitoring System in Michigan
I. Design, data and frequencies of selected dairy cattle diseases. Prev. Vet. Med. 8:103-114.


CHAPTER 2
LITERATURE REVIEW
The Transition Period of Dairy Cows
The transition period in dairy cows is defined as the last three weeks before
parturition to three weeks after parturition (Grummer, 1995). It is characterized by
tremendous metabolic and endocrine adjustments that the cows must experience from late
gestation to early lactation (Drackley et al., 2001). Perhaps the most important
physiological change occurring during this period is the decrease in dry matter intake
around parturition and the sudden increase in nutrients that cows need for milk production
(Drackley, 1999; Ingvartsen and Andersen 2000). As a result of these remarkable changes,
most of the infectious diseases and metabolic disorders occur during this time (Goff and
Horst, 1997b; Drackley, 1999). Milk fever, ketosis, retained fetal membranes (RFM),
metritis and displacement of the abomasum (DA) primarily affect cows within the first
two weeks of lactation (Drackley, 1999). Physical and metabolic stresses of pregnancy,
calving and lactation contribute to the decrease in host resistance during the periparturient
period (Mallard et al., 1998). During two weeks before and after parturition the T-cells
populations exhibit a significant decline, which contribute to the immunosuppession in
dairy cows at calving (Kimura et ah, 1999). This immunosuppression during the
periparturient period leads to increased susceptibility to mastitis and other infectious
diseases (Mallard et ah, 1998). Other diseases that are not clinically apparent during the
first two weeks of lactation (laminitis, ovarian cysts, endometritis) can be traced back to
insults that occurred during early lactation (Goff and Horst, 1997b).
4


29
myometrial activity between days 1 and 4 after calving, suggesting that uterotonic agents
are unlikely to promote placental expulsion. In addition, the levels of PGFM are higher in
cows with RFM than control cows (Burton et al., 1987; Risco et al., 1994b). Treatment of
cows with RFM with intrauterine tetracycline may reduce fertility (Youngquist and Braun,
1993). Parenteral administration of antibiotic agents is indicated in cases of sepsis
associated with RFM (Smith et al., 1998), however, the residues in milk must be
considered (Dinsmore et al., 1996).
Treatment of the uterine infections traditionally has been based on the use of local
and/or systemic antibacterial compounds. However, according to different trials, the
results have been controversial (Gilbert and Schwark, 1992; Youngquist, and Braun, 1993;
Pugh et al., 1994; Smith et al., 1998).
Uterine infections should be prevented by proper nutritional management during
the dry period, allowing cows to calve in an uncontaminated environment, and employing
strict sanitation if assistance is required during delivery (Youngquist and Braun, 1993;
Lewis 1997). Ultimately, the competence of the immune system has to be considered in
the prevention of the metritis complex. Uterine trauma, such as dystocia, manual removal
of RFM and intrauterine infusions, reduced the phagocytic activity of uterine and blood
neutrophils (Cai et al., 1994). Changes in immune function are consistent with changes in
estradiol and progesterone concentrations around calving and during the estrous cycle of
cows and ewes. Prostaglandins and other arachidonic acid metabolites might be important
mediators of resistance or susceptibility to uterine infections (Lewis 1997; Goff and Horst,
1997). Repeated doses of prostaglandin F2alpha beyond 7 days postpartum in cows with
metritis resultes in less acute response protein concentrations and lower diameter of


149
Table 5-3. Least squares means S.E.M. of mature equivalent (ME305) and actual
lactational milk yield (kg/lactation) in treated and control cows in different
Controls
Monensin
Primiparous
(ME305)
12268 109
12389 108
Multiparous
(ME305)
11096.4 121.3
11225 123
Primiparous
(Actual)
10592 95
10622 98
Multiparous
(Actual)
10623 97
10604 98


2
production, reduces production of methane and has a sparing effect on ruminal protein
digestion (Richardson et al., 1976; Yang and Russell, 1993; Nagaraja et ah, 1997; Becket et
ah, 1998; Duffield et ah, 1998a). These metabolic changes have been related with a decrease
in the incidence of some metabolic disease such as ketosis and fatty liver and an increase in
milk yield. Monensin, used as a controlled-release capsule, during the prepartum period,
decreased the acetate to propionate ratio, increased the concentration of blood glucose,
decreased serum NEFA concentrations, increased ruminal pH, decreased the ruminal butyrate
production and subsequently decreased the BHB concentration (Duffield et ah 1998a, 1998b;
Green et ah, 1999). Most of these studies were conducted in Canada and other countries,
demonstrating an improvement of the overall performance of lactating dairy cows.
Unfortunate, monensin is not allowed in lactating dairy cattle in the United States.
Citrus pulp is an energy concentrate by-product produced in subtropical regions, of
which south central Florida remain the largest area of production. Citrus pulp is a common
by-product used in diets for dairy and beef cattle in Florida and other southern states
(Arthington et ah, 2002). Citrus pulp is composed mostly by pectin, which is indigested by
mammalian enzymes, but can be rapidly fermented by ruminal microbes (Hall, 1997).
Diets based on citrus pulp consistently demonstrated an increase in milk fat content and
milk urea nitrogen, but not in milk yield, when compared with diets richer in starch
(Belibasakis and Tsirgogianni, 1996; Leiva et ah, 2000).
Pectin-fermenting bacteria are gram-negative monensin-resisting bacteria (Nagaraja
et ah, 1997; Stewart et ah, 1997). The positive effect of monensin has been demonstrated
on diets rich in starch (Nagaraja et ah, 1997). However, no trials have been conducted to
test the effect of monensin on transition cows fed diets based on citrus pulp. Since


16
approximately $335 per case (Dohoo and Martin, 1984; Erb et al., 1985; Bigras-Poulin et
al., 1990; Guard, 1994; Grdhn et al., 1998; Kelton et al., 1998).
Several risk factors related to MF have been identified. Breed, age and milk yield
are the most important risk factors for MF in dairy cattle. Jersey and Guernsey cows are
the most susceptible to MF; Holstein and Brown Swiss are moderately susceptible; and
Ayrshire and Milking Shorthorns are the least susceptible (Oetzel and Goff, 1999). The
incidence of MF increases with parity and with higher levels of milk production,
regardless of breed (Oetzel and Goff, 1999).
Most cows are in negative Ca balance during the early weeks of lactation because
more Ca leaves the body via milk, endogenous fecal loss, and urine than is absorbed from
the diet. This is because the intestinal mechanisms for absorbing calcium are not fully
adapted to lactation and also because dry matter intake is less than favorable (Oetzel and
Goff, 1999). Bone Ca mobilization is stimulated by a concerted effort of PTH and
l,25(OH)2D, but intestinal Ca absorption is controlled by l,25(OH)2D alone. During the
dry period, these mechanisms for replenishing plasma Ca are relatively inactive. Thus,
nearly all cows experience some degree of hypocalcemia during the first days after calving
as the intestine and bone adapt to lactation (Goff et al., 1996). The adaptation process
begins with dramatic increases in the plasma concentrations of PTH and l,25(OH)2D at
the onset of hypocalcemia. About 24 hours of l,25(OH)2D stimulation is required before
intestinal Ca transport is increased significantly. Bone Ca resorption (recruitment and
activation of osteoclasts) is not significantly increased until after about 48 hours of PTH
stimulation. In cows with MF, these adaptive processes can be even more prolonged
(Horst et al., 1994). Magnesium status is another factor influencing the risk of
hypocalcemia. Low blood magnesium levels can reduce PTH secretion from the


BCS (1-5)
135
Figure 4.1. BCS at dry-off and at calving in treated and control group in
primiparous and multiparous cows


175
Eiler, H. 1997. Retained placenta. Pages 340-348. In Current Therapy in Large Animal
Theriogenology. R. Youngquist. W.B. Saunders Company.
Emery, R. S. 1993. Energy needs of dry cows. Page 35 in Proc. Tri-State Dairy Nutr. Conf.
Ohio State Univ., Michigan State Univ., and Purdue Univ., Ft. Wayne, IN.
Erb, H. N. and Y. T. Grohn. 1988. Epidemiology of metabolic disorders in the periparturient
dairy cow. J. Dairy Sci. 71:2557-2571.
Erb, H. N., R. D. Smith, P. A. Oltenacu, C. L. Guard, R. B. Hillman, P. A. Powers, M. C.
Smith, and M. E. White. 1985. Path model of reproductive disorders and performance, milk
fever, mastitis, milk yield, and culling in Holstein cows. J. Dairy Sci. 68:3337-3349.
Fecteau, G., N. Sattler, and D. M. Rings. 1999. Abomasal physiology, and dilatation,
displacement, and volvulus. Pages 522-527 in Current Veterinary Therapy 4. Food Animal
Practice. J. Howard and R. Smith. W.B. Saunders Company.
Ferguson, J. M., D. T. Galligan, and N. Thomsen. 1994. Principal descriptors of body
condition score in Holstein cows. J. Dairy Sci. 77:2695-2703.
Fetrow, J. and K. Anderson. 1987. The economics of mastitis control. Compend. Contin.
Educ. Pract. Vet. 9:F103-F112
Fourichon, C., H. Seegers, N. Bareille, and F. Beaudeau. 1999. Effects of disease on milk
production in the dairy cow: a review. Prev. Vet. Med. 41:1-35.
Funk, D. A., A. E. Freeman, and P. J. Berger. 1987. Effects of previous days open, previous
days dry, and present days open on lactation yield. J. Dairy Sci. 70:2366-2373.
Geishauser, T., K. Leslie, D. Kelton, and T. Duffield. 1998. Evaluation of five cowside test
for use with milk to detect subclinical ketosis in dairy cows. J. Dairy Sci. 81: 438-443.
Geishauser, T., Leslie, K., Kelton, D., Duffield, T. 2001. Monitoring for subclinical ketosis in
dairy herds. Compendium Cont. Educ. Food Anim. 23:S65-S71.
Geishauser, T., K. Leslie, J. Tenhag, and A. Bashiri. 2000. Evaluation of eight cow-side
ketone tests in milk for detection of subclinical ketosis in dairy cows. J. Dairy Sci. 83:296-
299.
Gerloff, B.J. 2000. Dry cow management for the prevention of ketosis and fatty liver in dairy
cows. Vet. Clin. North Am. Food Anim. Pract. 16:283-292
Gerloff, B. J., and T. H. Herdt. 1999. Fatty liver in dairy cattle. Pages 230-233 in Current
Veterinary Therapy 4. Food Animal Practice. J. Howard and R. Smith. W.B. Saunders
Company.


152
20 -I 1 1 1 1 , , ,
T1 T2 T3 T4 T5 T6 T7 T8 T9 T10
DHIATest day
Figure 5.3. Test day milk production by treatment in multiparous cows