Citation
Vitamin E status in swine as affected by form or level of dietary vitamin E and/or by supplementation of vitamin A

Material Information

Title:
Vitamin E status in swine as affected by form or level of dietary vitamin E and/or by supplementation of vitamin A
Creator:
Anderson, Lee E., 1944-
Publication Date:
Language:
English
Physical Description:
ix, 71 leaves : ill. ; 29 cm.

Subjects

Subjects / Keywords:
Acetates ( jstor )
Blood ( jstor )
Fats ( jstor )
Gilts ( jstor )
Liver ( jstor )
Swine ( jstor )
Tissue samples ( jstor )
Tocopherols ( jstor )
Vitamin E ( jstor )
Vitamins ( jstor )
Animal Science thesis Ph. D
Dissertations, Academic -- Animal Science -- UF
Genre:
bibliography ( marcgt )
non-fiction ( marcgt )

Notes

Thesis:
Thesis (Ph. D.)--University of Florida, 1993.
Bibliography:
Includes bibliographical references (leaves 66-70).
General Note:
Typescript.
General Note:
Vita.
Statement of Responsibility:
by Lee E. Anderson.

Record Information

Source Institution:
University of Florida
Holding Location:
University of Florida
Rights Management:
Copyright [name of dissertation author]. Permission granted to the University of Florida to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Resource Identifier:
030376480 ( ALEPH )
30991190 ( OCLC )
AKB7343 ( NOTIS )

Downloads

This item has the following downloads:


Full Text











VITAMIN E STATUS IN SWINE AS AFFECTED BY FORM OR LEVEL OF
DIETARY VITAMIN E AND/OR BY SUPPLEMENTATION OF VITAMIN A















By

LEE E. ANDERSON, SR.


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

1993


UNIVERSITY OF FLORIDA LIBRARIES

































This dissertation is dedicated to my wife, Erma, and
children, Valerie, Lee Jr., Jenaya, Calvin, and Kelvin, and
to the memory of my parents, Florence Anderson, Myrtis
Stepherson and Leon Anderson, for their love, support and
encouragement.














ACKNOWLEDGMENTS


The author sincerely appreciates the efforts of all

members of his supervisory committee. Dr. Robert 0. Myer is

greatly appreciated for his understanding, guidance, advice

and companionship over the past several years. Sincere

thanks go to Dr. Joel Brendemuhl for his help and guidance

and his expertise and assistance during data collection.

The author is very appreciative of Dr. Lee McDowell's help

and advice, especially his assistance with acquiring the

vitamins. The advice, assistance and encouragement of Dr.

Jimmy Cheek and Dr. Joseph Conrad are deeply appreciated.

Thanks go to Dr. George Combs for his assistance. Without

the support of the above-mentioned individuals, this

dissertation would not have been possible.

Special gratitude is kindly extended to Dr. Jack Fry,

Dr. Roderick McDavis and Dr. Charles Kidd for their

encouragement and assistance during the course of this

program. Thanks are extended to Mr. Stephen Linda for the

valuable assistance he has given in the statistical analysis

of the data collected.

To his fellow graduate students, Maria Soler, Kim and

Kelly Sheppard, Dave Davis and Ken Mooney, the author

extends his thanks for their contributions and sacrifices in

iii








assisting him in this research. Special appreciation is

extended to Maria Soler and Kim Sheppard for their time and

involvement in trial 3.

Deep appreciation is extended to Alvin Boning for his

companionship and enthusiastic assistance in the laboratory.

Thanks go to Nancy Wilkinson for her assistance in the

laboratory. The author would like to take this opportunity

to thank Dane Bernis, Tom Crawford, Dennis Perry, Jack

Stokes, Dean Glicco and Rome Williams for their involvement

in the preparation of diets, care and management of the

experimental animals. Special appreciation is extended to

Leroy Washington, Larry Eubanks and Art Rogers for their

assistance in collecting tissue samples.

The author thanks Dr. Scot Williams and Hoffmann-La

Roche for providing the vitamins used in trials 2 and 3.

A special thanks is also extended to Mary Chambliss for

typing the manuscripts for review and publication.
















TABLE OF CONTENTS


Page

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

LIST OF TABLES. ............. ........................ vi

ABSTRACT. ............................................ viii

CHAPTERS

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

2 POTENCY OF VARIOUS VITAMIN E COMPOUNDS FOR
FINISHING SWINE ................................ 9

Introduction................................ .... 9
Experimental Procedures....................... 10
Results and Discussion......................... 14
Summary........................................ 25

3 THE EFFECT OF EXCESSIVE DIETARY VITAMIN A ON
PERFORMANCE AND VITAMIN E STATUS IN SWINE FED
DIETS VARYING IN DIETARY VITAMIN E............. 27

Introduction .................................. 27
Experimental Procedures......................... 28
Results and Discussion.......................... 32
Summary........................................ 42

4 EFFECT OF INJECTED VITAMIN A AND DIETARY
SUPPLEMENTATION OF VITAMIN E ON REPRODUCTIVE
PERFORMANCE AND TOCOPHEROL STATUS IN GESTATING
GILTS ........................................... 44

Introduction............................... .... 44
Experimental Procedures........................ 45
Results and Discussion......................... 48
Summary............................................. 59

5 GENERAL CONCLUSIONS........................... 63

REFERENCES.......................................... 66

BIOGRAPHICAL SKETCH......................... ........ 71














LIST OF TABLES


Table Page
2-1 Composition of finisher diet...................... 11

2-2 Performance of finishing pigs fed diets containing
various vitamin E compounds...................... 15

2-3 Vitamin E (a-tocopherol) concentrations in feed... 17

2-4 Adjusted serum vitamin E (tocopherol) concentrations
in finishing swine fed diets supplemented with
different vitamin E compounds..................... 19

2-5 Relative biopotency of vitamin E compounds (%).... 22

2-6 Adjusted tissue a-tocopherol concentrations in
finishing swine fed diets supplemented with
different vitamin E compounds..................... 23

3-1 Composition of diets (%) ......................... 29

3-2 Performance of growing-finishing swine fed diets
with different dietary levels of vitamin E
and vitamin A..................................... 33

3-3 Mean a-tocopherol concentrations in blood serum
due to dietary additions of vitamins E and A...... 34

3-4 Main means of serum a-tocopherol due to dietary
additions of vitamins E and A.................... 35

3-5 Main means of serum retinol due to dietary additions
of vitamins E and A............................... 38

3-6 Main means of tissue a-tocopherol concentrations due
to dietary additions of vitamins E and A.......... 40

3-7 Main means of tissue retinol concentrations due to
dietary additions of vitamins E and A............. 41

4-1 Composition of diet fed to gestating gilts........ 46








4-2 Mean reproductive response criteria of gestating
gilts given dietary additions of vitamin E and
injected with vitamin A.......................... 49

4-3 Mean serum a-tocopherol concentrations in gestating
gilts given dietary additions of vitamin E and
injected with vitamin A............................ 52

4-4 Main mean serum a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin
E and injected with vitamin A..................... 53

4-5 Mean serum retinol concentrations in gestating gilts
given dietary additions of vitamin E and injected
with vitamin A.................................... 54

4-6 Main mean serum retinol concentrations in gestating
gilts given dietary additions of vitamin E and
injected with vitamin A........................... 55

4-7 Mean tissue a-tocopherol concentrations of gestating
gilts given dietary additions of vitamin E and
injected with vitamin A.......................... 57

4-8 Main mean tissue a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin E
and injected with vitamin A....................... 60


vii














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

VITAMIN E STATUS IN SWINE AS AFFECTED BY FORM OR LEVEL OF
DIETARY VITAMIN E AND/OR BY SUPPLEMENTATION OF VITAMIN A

By

Lee E. Anderson, Sr.

August, 1993

Chairman: R. O. Myer
Major Department: Animal Science

Experiment one used 40 finishing pigs (80 kg) to

determine the potency of vitamin E compounds. Pigs were

divided among five nutritionally adequate diets supplemented

with DL-a-tocopherol, DL-a-tocopheryl acetate, D-a-

tocopherol, D-a-tocopheryl acetate or no vitamin E. Blood

and tissue samples were collected. Vitamin E forms

increased (P < .05) serum a-tocopherol concentrations by d 2

of the feeding period. Serum tocopherol in pigs fed acetate

forms remained elevated through out the study; serum

concentrations declined (P < .01) in pigs fed alcohol forms.

D-a-tocopheryl acetate resulted in highest serum and tissue

tocopherol. The potency of D-acetate form was greater for

swine than that predicted from bioassays with the rat.

Experiment two evaluated excessive dietary vitamin A on

vitamin E status and performance of growing-finishing pigs.


viii








Eighty-four pigs were fed corn-soybean meal based diets

supplemented with DL-a-tocopheryl acetate to provide 0, 15

or 150 IU of vitamin E/kg and with retinyl acetate to

provide 2,000 or 20,000 IU of vitamin A/kg of diet. Serum

and tissue tocopherol increased (P < .01) as dietary levels

of vitamin E increased. The data indicated that 20,000 IU

of vitamin A/kg of feed did not affect performance or serum

and tissue tocopherol.

In experiment three, 32 gilts were used to determine

the effects of vitamins A and E on reproductive performance

and on serum and tissue concentrations of vitamin E during

early gestation. Treatments consisted of corn-soybean meal

based diets supplemented with DL-a-tocopheryl acetate to

provide either 25 or 500 IU of vitamin E/kg of diet,

beginning d -7 prebreeding through d 25 of gestation. Half

of the gilts were injected with 350,000 IU of vitamin A

(retinyl palmitate) at d -7, again at breeding (d 0), and at

d 7 postbreeding. Reproductive performance was not affected

by treatment. Serum tocopherol increased (P < .01) with 500

IU of vitamin E. High (500 IU/kg) dietary vitamin E

increased tocopherol level (P < .01) in all tissues except

adipose. High vitamin A (350,000 IU) via injections had no

consistent effect on reproductive performance or on serum or

tissue concentrations of a-tocopherol or retinol.














CHAPTER 1
INTRODUCTION


Vitamin E was discovered in 1922 as a missing, needed

dietary factor (Brandner, 1971; Ullrey, 1981; Raacke, 1983;

McDowell, 1989). Vitamin E was isolated as alpha-

tocopherol. The name tocopherol means to bring forth

offspring (McDowell, 1989). George M. Calhoun, a professor

of Greek at the University of California, Berkeley, named

the new vitamin tocopherol in 1936 (tocos for childbirth,

phero to confer, and ol for alcohol) (Evans, 1962; Ullrey,

1981; Raacke, 1983).

It was recognized in 1920 that reproductive failure

occurred in rats consuming diets thought to be nutritionally

adequate. An unknown dietary factor, then called X and

later determined to be vitamin E, was deficient, which

resulted in fetal death and embryo resorption in the

laboratory rat (Evans, 1962; Mason, 1980; Diplock, 1985;

McDowell, 1989). Estrus and mating were normal, but fetuses

died and were resorbed unless the diet was supplemented with

small amounts of wheat germ, dried alfalfa leaves, or fresh

lettuce, which contained the deficient vitamin E (Evans,

1962; Mason, 1980; McDowell, 1989). Degeneration of the

germinal epithelium in male rats was prevented by








2

supplements of fresh lettuce (Mason, 1980). Other animal

species (cattle, sheep, mink, and chickens) were able to

reproduce without dietary vitamin E, but in each case their

offspring died prematurely (Brandner, 1971).

Vitamin E became known as the fertility vitamin.

Many studies were done to determine if vitamin E affected

reproduction in humans. In most cases vitamin E had little

or no effect (McDowell, 1989).

Vitamin E deficiency in swine results in reduced

reproductive efficiency, locomotor incoordination, muscular

and hepatic necrosis, fibrinoid degeneration of blood vessel

walls and muscular dystrophy (McDowell, 1977).

Vitamin E is a hydrophobic, peroxyl radical-trapping,

chain-breaking antioxidant found in the lipid fraction of

living organisms. Its principal function is to protect the

lipid material of an organism from oxidation (Machlin, 1980;

Burton et al., 1983; Diplock, 1985; McDowell, 1989; Coelho,

1991). Lipid peroxidation of membranes of cells and

cellular constituents can be very damaging. Damage may be

as simple as breaking a membrane and allowing leakage of

contents, or as complex as breaking a membrane containing

destructive enzyme systems. Hemolysis of red blood cells is

an example of relatively simple membrane breakage. Membrane

damage to lysosomes can be particularly devastating.

Lysosomes are sometimes called the "suicide bags" of the

cell, and when their membranes are broken they release








3

enzymes that hydrolyze tissue constituents and magnify

tissue damage (Tappel, 1962). Damage to the membrane of

such other cellular components as mitochondria and

microsomes, which contain 25 and 40 % unsaturated lipid,

respectively, have profound effects. In both microsomes and

mitochondria, vitamin E is the only known lipid antioxidant

(Tappel, 1962).

Selenium (Se) is a trace mineral that is known to spare

some of the requirement for vitamin E. Selenium is a

component of the enzyme glutathione peroxidase, which is a

selenoprotein containing four atoms of selenium per molecule

of protein (Scott, 1969; Draper, 1980). Glutathione is a

hydrogen donor. Vitamin E functions as a fat soluble

antioxidant, and selenium functions as a water soluble

antioxidant (Cunha, 1977). Vitamin E is the first line of

defense against peroxidation of fats in cells. If peroxides

are formed, selenium through the enzyme glutathione

peroxidase destroys the peroxides before tissue damage can

occur. Thus, selenium is considered the second line of

defense (Diplock, 1985; McDowell, 1989) and as a result,

both selenium and vitamin E are capable of preventing some

of the same nutritional diseases (McDowell, 1989). Vitamin

E can also reduce the selenium requirement by inhibiting

production of peroxides.

Pigs exhibiting clinical vitamin E and Se deficiency

signs have a pale, white discoloration of the skeletal









4

muscle, an enlarged, friable heart, associated with

hydropericardium, and sometimes intestinal edema and

hepatosis dietetica (Mahan and Moxon, 1980).

There are many factors that affect the bioavailability

of vitamin E. These include the form of vitamin E compound,

potency of compound, stability, absorption, other fat

soluble vitamins (e.g., retinol), mineral interactions, and

unsaturated fat. Bioavailability is defined as the

percentage of a drug or nutrient (in this case, vitamin E)

that enters the systemic circulation after administration

and the rate of entry into the general circulation for

distribution throughout the body as well as tissue

accumulation (Koch-Weser, 1974; "The American Heritage

Dictionary", 1982).

Eight forms of vitamin E are known to occur in nature,

four of which are referred to as tocopherols and four as

tocotrienols. They have been given Greek letter names to

distinguish them from one another (Diplock, 1985; NRC,

1988). The compounds differ in the placement of methyl

groups on the ring and the degree of saturation in the side

chain (McDowell, 1989).

Alpha-tocopherol is a yellow oil, soluble in certain

organic solvents. It is common practice to assay only this

isomer rather than all eight compounds because a-tocopherol

is the most biologically active, naturally occurring vitamin

E source (Ullrey, 1981).








5

DL-a-tocopherol has a potency of 1.1 IU/mg and its

acetate (DL-a-tocopheryl acetate) has a potency of 1 IU/mg

as determined by bioassays with rats. Activity of naturally

occurring a-tocopherol, D-a-tocopherol (also called RRR-

tocopherol) is 1.49 IU/mg and of its acetate, 1.36 IU/mg.

D-a-tocopherol is the most biologically active form (IU per

unit of weight; NRC, 1988).

Loss of vitamin E potency occurs in mixed feed from a

number of factors. The naturally occurring tocopherols have

relatively poor stability during processing, grinding,

pelleting, and storing at high temperatures or under moist

conditions. Vitamin E will also readily interact with other

ingredients in feed formulations (Adams, 1978; NRC, 1988).

More pigs are being raised in confinement without

access to pasture, which is an excellent source of vitamin

E. Heating and pelleting feed grains lower their vitamin E

values. The use of high moisture grain increases the need

for vitamin E supplementation due to the destruction of the

vitamin. Feeds formulated with fats containing high

quantities of unsaturated fatty acids are susceptible to

rancidity, which destroys vitamin E (Cunha, 1977). Malm et

al. (1976) reported that diets high in polyunsaturated fatty

acids increased vitamin E requirement and that pigs fed a

polyunsaturated fatty acid, low vitamin E diet throughout

the postweaning period resulted in some degree of red blood

cell destruction.








6

The alcohol form, a-tocopherol, is easily destroyed by

oxidation. Oxidative destruction of a-tocopherol is

accelerated by heat, light, moisture, unsaturated fats,

sulfates, nitrates and molds, and in diets containing

increased levels of copper, iron, zinc and manganese

(Ullrey, 1981; McDowell, 1989; Dove and Ewan, 1991; Mahan,

1991; Thompson, 1993). A more stable source of vitamin E is

a-tocopheryl acetate. Alpha-tocopheryl acetate is

chemically synthesized by esterification of a-tocopherol

with acetic acid. DL-a-tocopheryl acetate is the

international standard for vitamin E activity.

Vitamin E is fat soluble and as such its absorption is

associated with that of lipids. Vitamin E is absorbed in

the alcohol form. Vitamin E acetate is hydrolyzed to the

alcohol form in the small intestine prior to absorption.

Droplets of triglycerides are degraded by lipase and bile

into monoglycerides and free fatty acids, which form into

micelles. Micelles contain the lipid components including

the fat soluble vitamins. Vitamins are absorbed with the

fatty acids and monoglycerides. Triglycerides are re-formed

in the intestinal cell and packaged into chylomicrons.

Chylomicrons are absorbed into the lacteal ducts and carried

into the lymphatic system until they enter the general

circulation and are distributed to various tissues.

Factors interfering with digestion and absorption of lipid

affect the bioavailability of vitamin E.








7

Competition for absorption sites in the small

intestines among the fat soluble vitamins may affect

bioavailability of vitamin E. Vitamin A (retinol) may

interfere with both absorption and blood concentrations of

vitamin E. This has been demonstrated in chicks (Sklan and

Donoghue, 1982; Abawi and Sullivan, 1989) and rats (Blakely

et al., 1991). This effect appears to be due to increased

oxidation of vitamin E prior to the digesta reaching the

duodenum. This would result in vitamin E concentration

being lower at the major absorption sites in the upper small

intestine (Sklan and Donoghue, 1982). In this case, vitamin

E is oxidized at the expense of vitamin A. Erdman et al.

(1988) reported that vitamin E may protect vitamin A from

oxidation in the gastrointestinal tract and within cell

membranes. Reports that vitamin A toxicity in chicks has

been completely reversed with high dietary vitamin E

supplementation (Arnrich and Arthur, 1980) also indicate an

additional loss of vitamin E resulting in an increased need.

Young children, who were vitamin A deficient, absorbed more

vitamin A when given high supplemental levels of vitamin E

(Kusin et al., 1974) indicating that vitamin A may also

affect the availability of vitamin E. However, there is

very little or no research regarding the influence of

vitamin A on the vitamin E status of swine.

Recently there has been an interest in increasing

supplemental vitamin A levels via i.m. injections in








8

gestating gilts and sows. Extra vitamin A given just

before, during, and shortly after breeding has been reported

to improve reproductive performance in breeding swine (Brief

and Chew, 1985; Coffey and Britt, 1993). The elevation of

maternal plasma vitamin A is believed to improve embryonic

survival (NRC, 1988). The elevated vitamin A may also

affect bioavailability of vitamin E and/or its requirement.

Selection for increased growth rate and reproductive

performance increases dietary vitamin E requirements. In

addition, confinement rearing and feeding cereal-soybean

meal diets that vary considerably in vitamin E content, make

it important to insure that adequate levels of nutrients are

included in the diet. Fortification of diets adequately

supplemented with vitamins is extremely important in

optimizing performance under current production conditions.

More research is necessary to ascertain the significant

aspects of vitamin E and its enhancement or impediment on

performance under conventional swine production systems.

Therefore the focus of this manuscript is on the

bioavailability of vitamin E as affected by the type of

vitamin E compound fed, and the influence of vitamin A

supplementation on vitamin E status of growing-finishing

pigs and during early gestation of gilts.














CHAPTER 2
POTENCY OF VARIOUS VITAMIN E COMPOUNDS FOR FINISHING SWINE


Introduction


Vitamin E is an essential nutrient for normal growth,

health and reproduction in swine. Vitamin E requirement for

swine ranges between 10 and 22 IU/kg of diet (NRC, 1988).

Swine diets consisting mainly of corn and soybean meal

usually do not contain adequate amounts of vitamin E needed

to meet the pig's requirement (NRC, 1988). In addition, the

stability of all naturally occurring vitamin E forms are

very poor in mixed feed (Ullrey, 1981; Dove and Ewan, 1991;

Hidiroglou et al., 1992). Therefore, supplementation of

swine diets with a readily available form of vitamin E

assures that swine will receive the correct amount for

optimum performance.

Eight forms of vitamin E occur in nature (4

tocopherols, 4 tocotrienols). D-a-tocopherol has the

greatest biological activity (highest IU per unit weight;

NRC, 1988) but acetate and succinate forms are more stable

(Erdman et al., 1988). This experiment evaluated the

relative biopotencies of four forms of vitamin E (DL-a-

tocopheryl acetate, D-a-tocopheryl acetate, DL-a-tocopherol,

and D-a-tocopherol) when supplemented in the diet of

9








10

finishing swine. The concentration of a-tocopherol in blood

serum and tissue was used as an indicator of potency.


Experimental Procedures


Forty crossbred finishing pigs, 20 barrows and 20

gilts, with an average initial weight of 80 kg were randomly

assigned by sex to individual pens. Treatments were

randomly assigned to the pens such that each treatment

consisted of 8 pigs (4 barrows, 4 gilts). Treatments

consisted of the following supplemented vitamin E forms: DL-

a-tocopherol, DL-a-tocopheryl acetate, D-a-tocopherol, and

D-a-tocopheryl acetate. A negative control, which received

no supplemental vitamin E, was also included to give a fifth

treatment. Vitamin E forms used were pure forms supplied in

an unprotected oil solution (Sigma Chemical Co., St. Louis,

MO). Supplemental vitamin E was added to the diets such

that pigs consuming 3.2 kg of feed would received 200 IU per

day. Pigs were fed a corn-soybean meal finishing diet

formulated with a modified vitamin premix (exclusive of

vitamin E) and .1 ppm of added selenium. Diets were

otherwise formulated following NRC (1988) guidelines.

Composition of the corn-soybean meal basal diet is given in

Table 2-1. The pigs were fed the finisher diet for 28 days.

Prior to the start of this trial all pigs were fed a diet

that contained 22 IU of vitamin E/kg of diet. Feed and

water were available ad libitum throughout the experiment.











Table 2-1. Composition of finisher diet


Ingredient %, as fed



Ground corn 82.05
Soybean meal (48%) 15.00
Dynafos 1.70
Ground limestone 0.80
Salt 0.25
Trace mineral 0.10
Vitamin mixb 0.05
Se premix' 0.05



aProvided 200 ppm zinc, 100 ppm iron, 55 ppm manganese,
11 ppm copper, and 1.5 ppm iodine.

bProvided 2.2 mg riboflavin, 11 mg niacin, 9 mg
pantothenic acid, 150 mg choline chloride, 11 ug vitamin
B,2, 1.5 mg vitamin K, 2750 IU vitamin A, and 440 IU
vitamin D3 per kg of diet.


cProvided 0.1 ppm selenium.








12

Pigs were housed in an open-sided building with solid

concrete floors. Individual pig weights and feed

consumption were recorded biweekly. The trial was carried

out in the spring (April-May) of the year. Pigs were

managed according to acceptable management practices

throughout the experiment. Protocol for animal care had

been approved by the University Animal Use Committee.

Blood samples were collected by jugular vein puncture

from each pig on d 0, 1, 2, 7, 14, 21, and 28 of the feeding

period. Blood samples were centrifuged after collection,

and serum was frozen and stored at -200C until analyzed.

Feed samples were taken from the feeders on d 0, 5, 14 and

21, frozen and stored at -200C until analyzed for a-

tocopherol concentration. On d 29, the 20 barrows were

slaughtered, using accepted slaughter procedures, at the

University of Florida meats laboratory and tissue samples

collected. Tissue samples included liver, muscle

(rhomboideus and semimembranosus), back fat (10th rib area)

and leaf fat. Tissue samples were frozen following

collection and stored at -200C until analyzed.

Procedures used for the extraction and determination of

a-tocopherol in blood serum were as previously described

(Njeru et al., 1992). Procedures were similar to those used

by McMurray and Blanchflower (1979a,b) except in our study,

propanol was used in the serum extraction instead of

ethanol. Most of the vitamin E activity in serum and tissue








13

was assumed to be a-tocopherol (Ullrey, 1981). Extraction

of vitamin E from tissues and feed was done using a

procedure outlined by Chung et al. (1992). This procedure

was a modification of that of McMurray and Blanchflower

(1979b) and Hatam and Kayden (1979).

Alpha-tocopherol concentration was determined using 50

ul of the reconstituted sample (serum, tissue or feed)

injected onto a LiChrosorb SI 60 column (Hibar Fertigsaule

RT pre-packed column RT 250-4 E, Merck, Darmstadt, Germany)

250 mm x 4 mm I.D. and using a Perkin Elmer 550 terminal

(Perkin-Elmer Corp. Analytical Instruments, Norwalk, CT), a

Perkin Elmer ISS-100 auto sampler, and a Perkin Elmer Series

4 Liquid chromatograph pump. The mobile phase consisted of

900:99:1 HPLC grade iso-octane, tetrahydrofuran and acetic

acid. The detector was a Perkin Elmer LS-4 Fluorescence

Spectrometer with an excitation wavelength of 290 nm and an

emission wavelength of 320 nm. Data were collected by a

Perkin Elmer LCI-100 Laboratory Computing Integrator. Flow

rate was 1 ml/min. The retention time of a-tocopherol was

5.2 minutes. Alpha-tocopherol (Eastman Kodak Company,

Rochester, NY) was used as a standard, and sample peaks and

retention times were compared to those of the standards.

Standard concentration was calculated to give a peak of 250

or 500 ng. Alpha-tocopherol concentration of samples was

calculated by the external standard method. Spiked samples

were found to have a mean recovery rate of 97%.








14

Potencies of the various vitamin E compounds were

determined by comparing areas under the time curve (AUC)

within the serum, feed and tissue samples. Serum and tissue

concentrations were adjusted based on actual feed intake and

a-tocopherol levels in the feed. Alpha-tocopherol

concentrations reported were adjusted to a constant feed a-

tocopherol concentration (d 0 feed level). Serum and tissue

means were analyzed using the general linear model procedure

(SAS, 1988). Analysis of variance compared treatment

differences in serum and tissue tocopherol concentrations.

Analysis of covariance was also applied to the serum data

using baseline (d 0 serum tocopherol) data as a covariate.

Treatment means were compared using the least significant

difference multiple comparison procedure.


Results and Discussion


Growth rate of all pigs was good over the duration of

the 28 d study. Daily feed intake and feed-to-gain ratio

were not affected (P > .1) by supplementation of the vitamin

E sources (Table 2-2); however, a slight improvement in

growth rate (P < .09) was obtained in pigs fed the D-a-

tocopherol form compared to the negative control. Asghar et

al. (1991) reported improved growth rates in growing-

finishing pigs fed dietary levels of DL-a-tocopheryl acetate

at 100 IU/kg of diet compared to pigs fed 10 IU/kg of diet.

In contrast, Chung et al. (1992) found no difference in











Table 2-2. Performance of finishing pigs fed diets containing
various vitamin E compounds


Vitamin E sourcea
Item DL-a-TAC D-a-TAC DL-a-TOH D-a-TOH Neg.
control
No. of pigs 8 8 8 8 8
ADG, kg 1.05b 1.0b 1.09bc 1.18c 1.02b
ADF, kg 3.87 3.5 3.66 3.79 3.4

F/G 3.78 3.61 3.37 3.28 3.55

"DL-a-TAC = DL-a-tocopheryl acetate; D-a-TAC = D-a-
tocopheryl acetate; DL-a-TOH = DL-a-tocopherol; D-a-TOH =
D-a-tocopherol.

"Means within the same row with a different superscript
differ significantly (P < .09).








16

growth performance due to vitamin E source (encapsulated D-

a-tocopherol or DL-a-tocopheryl acetate) or level (16, 48

and 96 IU/kg) in trials with young, starting swine.

Alpha-tocopherol analysis of the diets containing the

vitamin E forms are reported in Table 2-3. The vitamin E

forms were included in the diet so that pigs would consume

62 IU of added vitamin E/kg of feed (72 IU/kg total).

However, there was considerable variation in analyzed levels

among the dietary treatments. Also, there was some

variation in feed consumption among treatments. Therefore,

data reported were adjusted based on analysis of diets and

mean treatment group feed consumption (Table 2-3).

The indicator used to determine biopotency of the

vitamin E compounds was the concentration of a-tocopherol in

serum and selected tissues. Bratzler et al. (1950) found

that plasma tocopherol concentration reflected level of

tocopherol ingested in trials with young pigs fed different

concentrations of tocopherol. They also observed increases

of tocopherol in various tissues. Other researchers

indicated that an animal's vitamin E status can be

determined by measuring a-tocopherol concentration in serum

and various tissue after oral administration: Baker et al.

(1986) with humans, Hidiroglou and McDowell (1987) with

sheep, and Jensen et al. (1990) and Asghar et al. (1991)

with pigs. Numerous studies have shown that dietary vitamin

E compounds are effective in elevating blood tocopherol










Table 2-3. Vitamin E (a-tocopherol) concentrations in feed


Treatment Sampling daya
(vitamin E source) 0 5 14 21

--------------IU/kg-------------
DL-a-tocopheryl acetate 94(72) 70(54) 64(49) 56(43)
D-a-tocopheryl acetate 113(72) 104(66) 74(47) 72(46)
DL-a-tocopherol 94(72) 24(17) 21(16) 13(10)
D-a-tocopherol 83(72) 22(19) 19(16) 15(13)
Neg. control 10 5 5 5


'Day samples were taken after start of trial. Samples
were taken directly from feeder then frozen until analyzed.
Numbers in parenthesis represent adjusted levels adjusted
to a constant IU/kg extrapolated from d 0 levels.








18

concentration, and also that blood tocopherol concentration

increased with increasing dietary level of vitamin E

(Hidiroglou et al., 1988; Jensen et al., 1988; Behrens and

Madere, 1991; Asghar et al., 1991; Chung et al., 1992).

As expected, mean serum concentrations of tocopherol at

d 0 (baseline) were similar across all treatments (Table 2-

4). All vitamin E compounds fed in this experiment

increased (P < .01) serum tocopherol concentration. The

increase in serum tocopherol concentration was rapid. The

increase started on d 1, grew further by d 2 (P < .01), and

plateaued by d 7. Horwitt et al. (1984) noted in a study

with humans that serum a-tocopherol concentrations were

increased at 8 to 24 hr after ingestion of various vitamin E

forms. Howard et al. (1990) with pigs weaned at 28 d,

depleted of vitamin E for the next 38 d, and then fed 30 IU

of supplemental vitamin E in the form of D-a-tocopheryl

acetate or DL-a-tocopheryl acetate per kg of diet noted a

rapid increase in blood a-tocopherol. Jensen et al. (1990),

in a study with pigs, 49 d old, also observed a rapid

increase in serum tocopherol concentrations after feeding

supplemental DL-a-tocopheryl acetate. In both of the above

swine studies, the first blood samples were taken 7 d after

the start of the feeding trial.

Serum tocopherol concentrations of pigs fed both

acetate forms were maintained beyond d 7; however, levels

dropped steadily in pigs fed the alcohol forms and were











Table 2-4. Adjusted serum vitamin E (tocopherol)
concentrations in finishing swine fed diets
supplemented with different vitamin E compounds


Vitamin E source"
Day DL-a-TAC D-a-TAC DL-a-TOH D-a-TOH Neg.
control
----------------- g/ml------------------

0 .8 .8 .8 .8 .9
Slb9 1.2b 1.3b .8c
2 1.3c 1.8b 1.4c 1.6bc .8d
7 1.5c 2.2b 1.4c 1.4c .6d
14 1.4C 1.8b i. i 1.3' .4d
21 1.4c 1.8b .6d .9d .3e
28 1.4c 1.7b .5d .5d .4d

Note: Each mean is based on eight observations. Adjusted to
constant intake of 72 IU/kg diet based on d 0 feed
analyses (Table 2-3). Day 0 serum values were not
adjusted.

"DL-a-TAC = DL-a-tocopheryl acetate; D-a-TAC = D-a-
tocopheryl acetate; DL-a-TOH = DL-a-tocopherol; D-a-TOH =
D-a-tocopherol.

c"Means within the same row with a different superscript
differ (P < .01).








20

lower (P < .01) on d 21 and 28 than the acetate forms. This

drop was probably due to poor stability of the alcohol forms

in the feed (Table 2-3). Degradation of vitamin E occurs

through oxidation, and is accelerated by light, alkali, heat

and trace minerals (Ullrey, 1981; Erdman et al., 1988; Dove

and Ewan, 1991; Hidiroglou et al., 1992). In the absence of

oxygen, tocopherols are relatively heat, light, and alkali

stable (Ullrey, 1981). Stability of a-tocopherol is

increased by acylation of the compound (Ullrey, 1981).

Acetate forms of vitamin E were found to be quite stable in

feed in our study. Acetate forms of vitamin E have also

been noted to be stable compounds by other researchers

(Harris and Ludwig, 1949a,b; Ullrey, 1981; Dove and Ewan,

1991; Chung et al., 1992). In general, serum tocopherol

concentrations observed in the present study were similar to

those of other studies in which pigs were fed diets

containing similar levels of added vitamin E (Jensen et al.,

1988; Asghar et al., 1991).

While all vitamin E forms evaluated rapidly increased

serum tocopherol concentrations, there was some evidence of

a slight difference in the rate of this increase. Average

serum concentration in pigs fed the DL acetate form was not

increased (P > .05) until d 2, whereas serum concentrations

of pigs fed the other compounds were increased (P < .05) on

d 1. Horwitt et al. (1984) found that D-tocopherol raised

blood a-tocopherol concentrations faster than the D- or DL-








21

a-tocopherol acetate forms in research done with humans.

D-a-tocopheryl acetate resulted in higher serum

tocopherol levels than the DL- acetate form. Relative to

DL-a-tocopheryl acetate, the D- form of the same compound

had an average biopotency of 146% (IU basis; Table 2-5) or

199% (weight basis; 146 X 1.36). Howard et al. (1990)

determined a relative biopotency of 218% (weight basis) for

D-a-tocopheryl acetate relative to the DL- form of the same

compound in trials with growing pigs. Thus it would appear

that the D- acetate form has a higher biopotency for swine

than that determined from the traditional rat fetal-

resorption bioassays. However, Ames (1979) presented

evidence that the commonly accepted conversion value of 1.36

may be too low in trials evaluating the relative biopotency

of several vitamin E compounds using the rat fetal-

resorption assay. Both alcohol forms in our study exhibited

similar biopotencies, and there was evidence during the

early portion of the study that these forms were slightly

more biopotent than DL-a-tocopherol acetate (Table 2-5).

Alpha-tocopherol in tissues in general followed a

similar pattern to that observed with serum (Table 2-6).

Overall, pigs fed any of the compounds had tissue tocopherol

concentrations higher than the negative control. Other

researchers have also found that adding vitamin E to the

diet increased tissue concentrations (Bratzler et al., 1950;

Jensen et al., 1990; Asghar et al., 1991).









22

Table 2-5. Relative biopotency of vitamin E compounds (%)


day day day day day day
Compound 1 2 7 14 21 28 Avg.

D-a-TAC 154 146 151 151 141 132 146
DL-a-TOH 122 110 123 124 32 83 99
D-a-TOH 124 114 118 140 46 73 103

Note: Based on serum values. DL-a-tocopheryl acetate = 100;
IU basis.






























C 4
a)

0 4-4
-H 4-4

CU
41 *H
(0 l






00a
4-
C 41


0 -0
5
4-1
r-1 C

0 0)

4 r-4
a
o o
u 0

0 -
o 4

0
10


'0 H0 U

I :
t3 Q) 0

4J -H 0





9 C
C -r4 C
* 4 >





(0


. .


a)
0-4



0 v

Sto

j >4
-a






* m
CU



1 : 0
UO (

>0
0 *



D
H0 Hn

Ot a)


OC 3

r. 0
4-40




S3
a ) (0



0 0)
0 U0
o rS
S-H '0



0 -3 1

a) a)





Q C0
-H -l
a)0


Q)
*P

4-)
Q)







0
0


I
0
4-i) 4

1 Q)






I
0
a






o


u
4 -
QU





C11







0
4a)





00
04

i 4-


II




U0
S-a
l




a
O


Q)
4-4
4-4
-l



4-
-1
a






*4-i
Q)
a)



04
4-1





0)
4-1
-H

(U

4-)
-H
:s

0



0 *
V
aQ)


H O

C CU


-rl

c-H
0 C
Q) 0
s *m
S U








24

Generally, acetate vitamin E forms resulted in greater

tissue tocopherol concentrations than alcohol forms, and the

difference was significant (P < .02) in the liver and

rhomboideus. The greater tissue levels reflected the

greater stability of the acetate forms in the feed over the

duration of the study. Liver tocopherol averaged 4.6 Ag/g

for the acetate forms and 2.6 Ag/g for the alcohol forms.

Between the two acetate forms, D-a-tocopheryl acetate

resulted in the same tissue tocopherol concentrations as the

DL- form, with exception of the semimembranosus in which the

D- form resulted in higher concentrations (P < .02). Tissue

tocopherol concentrations were similar for the two alcohol

forms.

Liver had the highest tocopherol concentration of all

tissues evaluated. The liver could be an indicator of

dietary vitamin E status or reflect immediate status.

Jensen et al. (1990) indicated that serum and liver a-

tocopherol concentrations reflected the short term vitamin E

status of the pig and that muscle and fat tissue

concentrations reflected the pig's long-term vitamin E

status.

The concentration of tocopherol in all tissues other

than the liver was similar (P > .10) in pigs fed the two

acetate forms, with the exception of semimembranosus as

noted above. Jensen et al. (1988), in trials with growing

pigs, also noted that the liver had the highest









25

concentration of tocopherol, followed by adipose tissue and

skeletal muscle. Also, Asghar et al. (1991), feeding

growing pigs DL-a-tocopheryl acetate at 100 IU/kg of diet,

observed higher tocopherol concentrations in the liver,

followed by the heart, lung and kidney.

In conclusion, all vitamin E compounds evaluated almost

immediately (by d 1) began to increase serum a-tocopherol

concentrations upon ingestion. Tissue a-tocopherol

concentrations were reflective of serum concentrations.

Vitamin E acetate forms fed to finishing pigs resulted in

higher serum and tissue tocopherol concentrations than the

alcohol forms due to their greater stability in mixed feed.

Among the two acetate forms evaluated, the D- form had a

greater biopotency for swine than that determined by

traditional assays.


Summary


Relative biopotencies of four chemical forms of vitamin

E supplemented in diets of finishing swine for 28 d were

evaluated. Forty crossbred pigs (80 kg), individually

penned, were divided equally among five treatments.

Treatments consisted of corn-soybean meal based diets

supplemented with DL-a-tocopherol, DL-a-tocopheryl acetate,

D-a-tocopherol or D-a-tocopheryl acetate. A treatment

without vitamin E supplementation (negative control) served

as the fifth treatment. Each compound was supplemented at








26

62 IU/kg of diet. Blood samples were collected on d 0, 1,

2, 7, 14, 21, and 28. On d 29, half of the pigs were

slaughtered to obtain tissue samples. Feed samples, taken

from feeders, were collected on d 0, 5, 14, and 21. All

vitamin E forms fed increased (P < .05) serum a-tocopherol

concentration by d 2 and the concentration peaked by d 7.

Serum tocopherol concentrations in pigs fed either acetate

form remained elevated beyond d 7; serum concentrations

steadily declined and were lower (P < .01) on d 21 and 28 in

pigs fed either alcohol form in comparison to acetate-fed

pigs. The decrease was probably a reflection of reduced

stability of the alcohol forms in the feed; the acetate

forms were found to be stable in the feed over the 28 d

study. Dietary supplementation of D-a-tocopheryl acetate

resulted in the highest serum tocopherol concentrations

throughout the study, compared to concentrations obtained

for pigs fed the other compounds. A similar trend was

observed in tissue (liver, back fat, leaf fat,

semimembranosus, rhomboideus) tocopherol concentrations as

with serum concentrations, with the liver having the highest

concentration. The biopotency of D-a-tocopheryl acetate for

swine appears to be greater than predicted from traditional

bioassays with rats.














CHAPTER 3
THE EFFECT OF EXCESSIVE DIETARY VITAMIN A ON PERFORMANCE
AND VITAMIN E STATUS IN SWINE FED DIETS
VARYING IN DIETARY VITAMIN E


Introduction


Both vitamins A and E are fat soluble vitamins. There

is evidence that high dietary vitamin A may interfere with

both vitamin E absorption and blood a-tocopherol

concentration. High dietary vitamin A reduced absorption of

a-tocopherol in trials with chicks (Sklan and Donoghue,

1982). Abawi and Sullivan (1989) noted decreased plasma

vitamin E concentration when chicks received high (100,000

IU/kg) levels of dietary vitamin A. Blakely et al. (1991)

also reported that high dietary vitamin A (100 times

requirement) plus high levels of beta carotene (480 mg/kg of

diet) reduced plasma vitamin E concentration by 77% in

rats.

Limited research is available on the effect of dietary

vitamin A on vitamin E status in pigs. Our study was done

to evaluate the effect of excessive dietary vitamin A on

performance and on serum and tissue concentrations of a-

tocopherol of growing-finishing pigs fed diets supplemented

with varying levels of vitamin E.











Experimental Procedures


Eighty-four crossbred pigs with an average initial

weight of 26 kg were divided by sex, weight and litter

origin into pens of two pigs each (1 barrow, 1 gilt). Each

pen was assigned to one of six dietary treatments within

each of seven replications. The treatments for the 2 x 3

trial consisted of a basal corn-soybean meal diet

supplemented with DL-a-tocopheryl acetate (Hoffmann-La Roche

Inc., Nutley, NJ) at levels of 0, 15, or 150 IU/kg, and

retinyl acetate (Hoffmann-La Roche Inc., Nutley, NJ) at

levels of 2,000 or 20,000 IU/kg of diet. Pigs were fed a

grower diet (Table 3-1), formulated to meet NRC (1988)

requirements (except for vitamins A and E), until they

reached an average body weight of 57 kg and continued on a

finisher diet (Table 3-1) with the same treatment until they

reached an averaged body weight of 107 kg. The vitamin

premix used did not contain vitamins A and E. Pigs were

given feed and water ad libitum. Pigs were housed in an

open-sided building in pens with solid concrete floors.

Pigs were weighed at the start of the feeding phase and

biweekly thereafter. Average daily weight gains, feed to

gain ratios, and average daily feed intakes were determined.

Prior to the start of the study the pigs were fed a nursery

diet that contained 2750 IU of added vitamin A and 22 IU

added vitamin E/kg of feed. The trial was carried out










Table 3-1. Composition of diets


Ingredient Grower Finisher



Ground corn 75.00 82.05
Soybean meal (48%) 22.00 15.00
Dynafos 1.70 1.70
Ground limestone .80 .80
Salt .25 .25
Trace mineral premix" .10 .10
Vitamin premix .10b .05c
Se premixd .05 .05



aProvided 200 ppm zinc, 100 ppm iron, 55 ppm manganese,
11 ppm copper, and 1.5 ppm iodine.

bProvided 4.4 mg riboflavin, 22 mg niacin, 18 mg
pantothenic acid, 300 mg choline chloride, 22 ug
vitamin B,2 3 mg vitamin K, and 880 IU vitamin D3 per
kg of diet.

cProvided 2.2 mg riboflavin, 11 mg niacin, 9 mg
pantothenic acid, 150 mg choline chloride, 11 ug
vitamin B,, 1.5 mg vitamin K, and 440 IU vitamin D3 per
kg of diet.


dProvided .1 ppm selenium.








30

during late spring and early summer (March through June).

Pigs were managed following acceptable care and management

practices. Protocol for animal care had been approved by

the University Animal Use Committee.

Blood samples were collected by jugular venipuncture

from each pig at the start (d 0) and on d 3, 7, 21, 35, 63,

and 77 of the feeding period thereafter. Blood samples were

shielded from direct sunlight. Blood samples were

centrifuged after collection, serum harvested, frozen and

stored at -200C until analyzed. During storage, blood

samples were covered with foil to avoid exposure to light.

Upon termination of the feeding phase, one pig (barrow)

per pen was slaughtered, following accepted slaughter

procedures, at the University of Florida meats laboratory

and tissue samples collected. Tissue samples included

liver, leg (semimembranosus) and neck (rhomboideus) muscle,

back fat (10th rib area) and leaf fat. Tissue samples were

frozen following collection and stored at -200C until

analyzed.

Vitamin E (a-tocopherol) was extracted from serum

samples using the procedure described by McMurray and

Blanchflower (1979a) with modifications described by Njeru

et al. (1992). Extraction of vitamin E from tissue and feed

samples was done using the procedure of Chung et al. (1992).

This procedure was similar to that of McMurray and

Blanchflower (1979b) and Hatam and Kayden (1979) with








31

modifications (Njeru et al., 1992). Alpha-tocopherol was

determined by injecting 20 ul of the reconstituted sample

(serum, or tissue) into an HPLC (Anderson et al., In Press).

Alpha-tocopherol concentration of samples was calculated

from the known concentration of standards. Spiked samples

of a-tocopherol were found to have a mean recovery rate of

97 3%.

Vitamin A was extracted from serum and tissues as

described by Chew et al. (1991) and Mooney (1992).

Extraction procedures were performed under dark conditions

with either yellow filtered or subdued light. Vitamin A was

assayed and determined by the method of Mooney (1992). The

only modifications were that HPLC prepared samples were

eluted using an 80:20 (vol/vol) mixture of iso-octane:

tetrahydrofuran with 1% acetic acid and retention time was

approximately 10.5 min. All trans retinol (Sigma Chemical

Co., St. Louis, MO) standards were prepared and used to

determine concentration of samples. Several liver samples

were spiked to determine the recovery rate and validate the

extraction procedure. Recovery rate of retinol was found to

be 100 4.7%.

Data collection included serum and tissue

concentrations of vitamin A and E and performance data (body

weight gain, feed-to-gain and feed intake). Tissue data

were log transformed prior to analysis to improve

homogeneity of variance. A univariate repeated measures








32

ANOVA was performed on serum data. Data were analyzed using

the GLM procedure of SAS (1988). Orthogonal polynomial

contrasts were performed to compare treatment means.


Results and Discussion


Growth performance data of the pigs in this study are

summarized in Table 3-2. Pigs grew well on all dietary

treatments (NRC, 1988). An increase in average daily gain

of pigs approached significance (P = .15) in linear fashion

as dietary vitamin E increased; feed to gain was not

affected (P > .1). Dietary vitamin A levels had no effect

on pig performance (P > .1). Hoppe et al. (1992) fed pigs

54 IU of supplemental vitamin E in combination with 5,000,

10,000, 20,000 or 40,000 IU of retinol/kg of diet and also

observed no differences in pig performance. Other

researchers have noted similar results in the chick (Sklan

and Donoghue, 1982; Abawi and Sullivan, 1989) and rat

(Blakely et al., 1991).

Serum a-tocopherol concentrations are reported in

Tables 3-3 and 3-4. Serum tocopherol concentrations were

affected by dietary a-tocopherol on all sampling days except

d 0 (Table 3-4). When no supplemental vitamin E was added

to the diet, there was a steady decline in serum tocopherol

concentration from d 0 to d 77 (Table 3-4). When the diet

was supplemented with 15 IU of a-tocopheryl acetate per kg

of diet, serum tocopherol concentrations declined (P < .05)











Table 3-2. Performance of growing-finishing swine fed diets
with different dietary levels of vitamin E and
Vitamin A


Added
Vit. E, Added vitamin A, IU/kg
IU/kg 2,000 20,000 Mean SE
-- ----------Avg. daily gain, kg-----------------

0 .91 .91 .91
15 .92 .96 .94
150 .96 .92 .95

Mean .93 .94 .01
SE .02
-------------------Avg. daily feed, kg---------------

0 3.02 3.23 3.13
15 3.10 3.11 3.11

150 3.17 3.08 3.12
Mean 3.09 3.14 .05
SE .09
--------------------Avg. feed/gain --- -----------

0 3.33 3.56 3.44
15 3.39 3.25 3.32

150 3.29 3.32 3.30

Mean 3.34 3.38 .07
SE .08

Note: Seven pens per treatment with 2 pigs per pen.

'P>F:E = .19, A = .75, E*A = .22, E linear = .15

bP>F:E = .97, A = .52, E*A = .26, E linear = .94

cP>F:E = .41, A = .67, E*A = .26, E linear = .36











Table 3-3. Mean a-tocopherol concentrations in blood serum
due to dietary additions of vitamins E and A


Vit. E and A, IU/kcg
Days 0/2b 0/20b 15/2 15/20 150/2 150/20 SE
----------------------g/ml-----------------

0 1.14 1.17 1.18 1.24 1.03 1.25 .11
3 .60 .66 .75 .75 2.75 2.04 .09
7 .55 .59 .92 1.02 3.18 2.92 .11
21 .37 .53 .75 .84 3.15 2.99 .10
35 .39 .45 .87 .78 3.18 2.99 .09
63 .31 .32 .99 .78 3.29 3.24 .08
77 .33 .31 .98 .70 2.84 3.02 .11

Note: Each mean is based on 14 observations.

aVitamin E linear effect P < .01 all days except d 0.
E x A P < .01 d 3 and 21 only.


b2 = 2,000; 20 = 20,000 IU of retinyl acetate.











Table 3-4. Main means of serum a-tocopherol due to dietary
additions of vitamins E and A


Vit. Ea, IU/kg Vit. Ab, IU/kg
Days 0 15 150 SE 12,000 20,000 SE
------------------ig/ml----------------------

0 1.15 1.21 1.14 .07 1.11 1.22 .06
3 .63 .75 2.40 .06 1.37 1.15 .05
7 .57 .97 3.05 .08 1.55 1.51 .06
21 .45 .79 2.92 .07 1.42 1.35 .06
35 .41 .83 3.08 .06 1.48 1.40 .05
63 .31 .88 3.26 .05 1.53 1.45 .04
77 .31 .84 2.93 .08 1.38 1.34 .06

Note: Each mean is based on 28 or 42 observations.

"Vitamin E linear effect P < .01 all days except d 0; E
quadratic P < .02, d 63 and 77 only.

bVitamin A effect P < .01, d 3 only.








36

from their initial concentration by d 3 and then stabilized

throughout the remainder of the trial (Table 3-4). Serum

tocopherol concentration increased (P < .01) with the

highest vitamin E supplementation level by d 3 and continued

to increase (P < .01) to d 7 after which serum concentration

was maintained throughout the study (Table 3-4). Serum

tocopherol was highest (P < .01) at the highest level of

vitamin E supplementation on all days except d 0. Overall,

as dietary levels of supplemental vitamin E increased, serum

concentration of tocopherol also increased (linear; P <

.01). Other studies have shown dietary vitamin E compounds

are effective in increasing blood tocopherol and that blood

concentrations increased with increasing dietary vitamin E

(Jensen et al., 1988; Asghar et al., 1991 and Anderson et

al., In Press).

Serum tocopherol concentrations due to supplementing

the diet with vitamin A at 2,000 or 20,000 IU/kg of diet are

summarized in Table 3-4. Pigs fed the low level of vitamin

A tended to have higher concentrations of serum tocopherol

than pigs fed the high level of vitamin A on all blood

sampling days, but was significant only on d 3 (P < .01).

Among the three dietary levels of vitamin E, the highest

dietary vitamin E tended to be decreased by the high dietary

vitamin A to the greatest extent, but was significantly

lower (P < .01) on d 3 and d 21 only (Table 3-3). Although

significant differences were noted above, the magnitude of










these changes was very small. Therefore vitamin A level of

20,000 IU/kg had only a minimal effect on blood tocopherol

in swine. Weaver et al. (1989), in a trial with young

starting pigs, observed that high dietary levels of vitamin

A (9,900 and 16,500 IU/kg) fed with 33 IU of added vitamin

E/kg tended to lower plasma tocopherol concentrations

slightly. In trials with chicks and rats, however, a

decrease in blood tocopherol was noted upon feeding diets

with excessive vitamin A. In these trials, very high

vitamin A concentrations (> 100,000 IU/kg of diet) were fed

(Sklan and Donoghue, 1982; Blakely et al., 1991). This

negative effect is thought to be attributed to competition

for absorption sites in the small intestine or enhanced

oxidation of tocopherol prior to tocopherol reaching the

small intestine (Sklan and Donoghue, 1982).

Although some differences were observed on individual

sampling days, supplemental vitamin A had no effect (P >

.10) on serum retinol (Table 3-5). Differences that were

noted were very small and within normal values for serum

retinol usually found in the pig (Kaneko, 1980). Blood

retinol was also not affected by dietary supplementation of

various levels of vitamin A in studies done by other

researchers (Abawi and Sullivan, 1989; Blakely et al., 1991;

Hoppe et al., 1992).

With two exceptions, there was no effect (P > .10) on

serum retinol due to supplementation of vitamin E at any of










Table 3-5. Main means of serum retinol due to dietary
additions of vitamins E and A


Vit. Ea, IU/kg Vit. Ab, IU/kg
Days 0 15 150 SE l 2,000 20,000 SE
--------------------g/ml---------------

0 .34 .34 .39 .02 .31 .41 .02
3 .31 .33 .35 .01 .30 .35 .01
7 .34 .32 .35 .01 .33 .34 .01
21 .34 .36 .35 .01 .35 .35 .01
35 .37 .34 .32 .01 .35 .34 .01
63 .39 .35 .37 .01 .38 .36 .01
77 .47 .49 .46 .02 .52 .44 .02

Note: Each mean is based on 28 or 42 observations.

'Vitamin E linear effect P < .05 on d 3 and 35 only.

bVitamin A effect P < .01 on d 3 and 77 only.








39

the dietary levels evaluated in this study. Increasing

dietary vitamin E (Table 3-5) resulted in higher (P < .02)

retinol serum concentration on d 3 in pigs fed the highest

supplemental vitamin E, but also resulted in the lowest (P <

.02) serum retinol concentration on d 35. Although both

were significant, the differences were small. Weaver et al.

(1989) in trials with growing pigs also noted that plasma

vitamin A was not affected by dietary level of vitamin E.

Tissue a-tocopherol increased (linear; P < .001) as

dietary vitamin E increased (Table 3-6). As dietary vitamin

E increased from 15 to 150 IU per kg of diet, tissue

tocopherol increased by at least a factor of 2 or more in

all tissues evaluated. This finding is in agreement with

others who have observed similar responses in pigs (Jensen

et al., 1988; Asghar et al., 1991). Among the tissues

sampled, the highest concentration of a-tocopherol due to

treatment was found in the adipose tissue followed by liver

and muscle tissue, respectively (Table 3-6).

High supplementation of vitamin A had no effect (P >

.1) on tocopherol concentration in any of the tissues

studied (Table 3-6). The retinol concentration in the liver

was greatly enhanced (P < .001) when pigs were fed the high

level of vitamin A (Table 3-7). Retinol concentration in

other tissues (back fat, leaf fat, rhomboideus and

semimembranosus) was not detected and are not reported.

Dietary vitamin E had no effect (P > .10) on liver retinol











40



>4
H W(N I N N ml
Q) .



0 o






0 0


C I
O ** IO
H0 I V (N 'c ?





it > 0

--T 0 0 ( 0 O O
0 I



O > 0 L n







U r i n M O
0 W a)






0 0
r0 (




o) I 0



L0 4-4 H O *

o H H H 4 o
0 0
41U) 4 0









10 1 4 -J c O U
M 4-1 a)
SZ0 '0 0-












0 4
* r M U) 'O I





Ul C 0
I -4 r














fl)O Q ) Q) 0 '4-4 4-4 5
Sg 4 4-4 Q)
0 r 0o u o > u

U E)-,Qi 0 V
E- 0)
'0-H Qr 'a4) 4








41

Table 3-7. Main means of tissue retinol concentrations
due to dietary additions of vitamins E and A

Vit. E, IU/kq Vit. Aa, IU/kg
Tissue 0 15 150 SE 12,000 20,000 SE
---------------------------- /--------------------

Liver 503 470 496 44 103 876 36

Note: Each mean is based on 14 or 21 observations.

aVitamin A effect P < .001.








42

concentration (Table 3-7). Hoppe et al. (1992) found that

liver retinol was linearly related to dietary retinol in a

trial involving young, growing pigs as was also observed in

the present study. They also found that 10,000 IU of

dietary retinol did not affect tissue a-tocopherol other

than the heart in which a-tocopherol was slightly depressed.

However, Pudelkiewicz et al. (1964) feeding vitamin A

depleted chicks 0, 1,453, 14,535, 1,453,488 and 14,534,883

IU of vitamin A acetate/kg of diet observed a marked decline

in tocopherol concentration in liver tissue at the highest

dietary vitamin A levels.

In conclusion, there was no consistent evidence that

excessive dietary vitamin A (20,000 IU/kg of diet) affected

growth performance or serum or tissue a-tocopherol in

growing-finishing pigs fed diets supplemented with varying

levels of vitamin E. Likewise serum retinol was not

affected by dietary vitamin E.


Summary


A 2 X 3 factorial experiment was conducted to evaluate

excessive dietary vitamin A on vitamin E status and

performance of growing-finishing pigs fed diets supplemented

with varying levels of vitamin E. Treatments consisted of

corn-soybean meal based diets supplemented with DL-a-

tocopheryl acetate to provide 0, 15 or 150 IU added vitamin

E/kg and with retinyl acetate to provide 2,000 or 20,000 IU








43

vitamin A/kg of diet. The trial involved 84 crossbred pigs

(26 kg) divided by sex, weight, and genetic background into

pens of two pigs each. Treatment was assigned at random to

pens within each of seven replications. Pigs were fed

grower diets (.75% lysine) until they reached 57 kg average

weight and were then switched to finisher diets (.60%) until

107 kg. Serum was collected on day 0, 3, 7, 21, 35, 63, and

77 of the feeding period. Tissue samples (liver, muscle,

back fat and leaf fat) were collected from one pig (barrow)

in each pen at the end of the feeding phase. Overall

average daily gain and feed-to-gain were .93 kg and 3.35

respectively, without treatment differences (P > .1).

Excessive dietary vitamin A had no effect (P > .1) on serum

retinol concentrations except d 3 in which there was a small

(P < .01) increase. Serum tocopherol was increased (P <

.01; linear) by d 3 with dietary vitamin E supplementation

and was maintained (P < .01) throughout the feeding period.

High dietary vitamin A resulted in a small but significant

(P < .01) decrease in serum tocopherol on d 3; serum

tocopherol concentrations were not affected on other days.

Tissue tocopherol was increased (P < .001; linear) as

dietary vitamin E increased from 15 to 150 IU/kg. Liver

retinol increased (P < .001) by a factor of eight. No

consistent evidence was found that high dietary vitamin A

interfered with performance or with serum or tissue

tocopherol in growing-finishing swine.















CHAPTER 4
EFFECT OF INJECTED VITAMIN A AND DIETARY SUPPLEMENTATION OF
VITAMIN E ON REPRODUCTIVE PERFORMANCE AND TOCOPHEROL STATUS
IN GESTATING GILTS


Introduction


Supplemental vitamin A and/or B-carotene given via

injection just before and/or shortly after breeding appears

to enhance reproductive performance of gilts and sows (Brief

and Chew, 1985; Coffey and Britt, 1993). There is evidence

that high dietary vitamin A may interfere with both vitamin

E absorption and blood a-tocopherol concentrations. High

dietary vitamin A reduced absorption of a-tocopherol in

trials with chicks (Sklan and Donoghue, 1982). Abawi and

Sullivan (1989) noted a decrease in plasma vitamin E

concentrations when depleted chicks were administered high

(100,000 IU/kg) levels of dietary vitamin A. Blakely et al.

(1991) also reported that high levels of vitamin A (100

times requirement) plus high levels of B-carotene (480 mg/kg

of diet) reduced vitamin E plasma concentration by 77% in

rats.

This study was conducted to evaluate the effect of

injecting vitamin A just before, during and shortly after

breeding, and dietary supplementation of vitamin E on

reproductive performance and on blood and tissue

44








45

concentrations of a-tocopherol during early gestation of

gilts.


Experimental Procedures


The trial was a 2 x 2 factorial design and involved 32

(7 to 8 month old) crossbred gilts. The gilts were divided

into pens of 8 gilts each. Gilts used in this study were

from a previous trial that involved the feeding of diets

supplemented with either 2,000 (L) or 20,000 (H) IU vitamin

A/kg of diet. This was taken into consideration in the

allotment of gilts to treatment (4L and 4H per pen). Each

pen was randomly assigned to one of four treatments.

Treatments consisted of a basal corn soybean meal diet

(Table 4-1) supplemented with DL-a-tocopheryl acetate

(Hoffmann-La Roche Inc. Nutley, NJ) at levels of either 25

or 500 IU/kg of diet. Gilts were fed experimental diets

beginning 7 d prior to breeding through d 25 of gestation.

Half of the gilts were given three injections (i.m. in the

neck) of 350,000 IU each of vitamin A (vitamin A palmitate

Hoffmann-La Roche Inc. Nutley, NJ); the other half were

injected with vehicle only. The gilts were injected at 7 d

prebreeding (d -7), at breeding (d 0) and 7 d postbreeding

(d 7). Gilts were fed 1.9 kg of feed/hd once daily and

given free access to water. Gilts were housed in an open-

sided building with solid concrete floors. Gilts were

checked twice daily for estrus during the trial and doubled











Table 4-1. Composition of diet fed to gestating gilts


Ingredient


Ground corn 84.66
Soybean meal (48%) 12.40
Dynafos 1.54
Ground limestone .65
Salt .50
Trace mineral .10
Vitamin premixb .10
Se premixc .05



'Provided 200 ppm zinc, 100 ppm iron, 55 ppm manganese,
11 ppm copper, 1.5 ppm iodine, and 1 ppm cobalt.

bProvided 4.4 mg riboflavin, 22 mg niacin, 18 mg
pantothenic acid, 300 mg choline chloride, 22 ug vitamin
B,, 3 mg vitamin K, 880 IU vitamin D3, and 4000 IU
vitamin A per kg of diet.


'Provided .1 ppm selenium.








47

mated on their second or third observed estrus to duroc x

hampshire x yorkshire boars.

Blood samples were collected by jugular vein puncture

from each gilt on d -7, 0, 7 and 24 of gestation to monitor

the serum a-tocopherol (vitamin E) and retinol (vitamin A)

concentrations. Blood samples were covered with foil to

prevent exposure to direct sunlight, taken to the

laboratory, centrifuged and the serum harvested. Serum was

stored at -200C until analyzed for a-tocopherol and retinol

concentration. During storage, serum samples were covered

with foil to prevent exposure to light.

Gilts were slaughtered, following accepted slaughter

procedures, on d 25 of gestation at the University of

Florida meats laboratory. Reproductive tracts were

immediately removed and refrigerated for later counting of

corpora lutea (CL) and recovery of embryos. Tissue samples

were also collected which consisted of endometrium, embryo,

ovary, uterus, liver, leaf fat, back fat and muscle

(semimembranosus and rhomboideus). Tissue samples were

stored at -200C until analyzed for a-tocopherol and retinol

concentrations.

The trial was carried out during the summer and early

fall (July through October). Pigs were managed following

acceptable care and management practices throughout the

study. Protocol for animal care had been approved by the

University Animal Use Committee. Three pigs from different









48

treatment groups were eliminated from the study due to

death, lameness and failure to conceive.

Alpha-tocopherol was extracted from serum and tissues

using procedures as described earlier (Anderson et al., In

Press). Alpha-tocopherol concentration was determined by

injecting 10 ul of the extracted sample (serum and tissue)

into the HPLC system. Retinol was extracted, assayed and

concentration determined from the serum and tissues by the

method as described previously (Mooney, 1992).

Experimental data included serum and tissue

concentrations of a-tocopherol and retinol, and reproductive

performance. Tissue data was log transformed prior to

analysis to improve homogeneity of variance. A univariate

repeated measures ANOVA was performed on serum data (SAS,

1988). Data were analyzed as a 2 X 2 factorial design with

the factors being dietary vitamin E level and whether or not

gilts were injected with vitamin A.


Results and Discussion


Levels of vitamin E used were chosen to reflect NRC

(1988) requirement and to give a very high level. The

injected level for vitamin A was chosen because it was

thought to be the upper limit that would elicit a response

and not be toxic to the gilts (also some evidence of this

dosage was being used).

Reproductive performance data are summarized in Table









49

Table 4-2. Mean reproductive response criteria of gestating
gilts given dietary additions of vitamin E and injected
with vitamin A


Item

No. of CL
No. of Embryob
Embryo wt., g
Ovary wt., g
Uterus wt., kg


Vit. E, IU/kg and Vit. A inji.
25/No 25/Yes 500/No 500/Yes

14 13 15 15
12 13 12 15
9 9 7 8
13 13 14 15
3 2 2 2


SE

.76
1.13
.98
.90
.22


Note: Each mean is based on 7 or 8 observations.

"Three injections of 350,000 IU each.

bEffect of vitamin E (P = .14), effect of
vitamin A (P = .13).








50

4-2. Although none of the differences noted with

reproductive data were significant (P > .1), due to the

small number of gilts per treatment and the inherent nature

of swine reproductive data, nevertheless, some positive

trends were observed due to treatment. Gilts receiving the

high vitamin E and high vitamin A treatment had larger (P =

.16) litters than gilts given other treatments. Embryonic

survival was 86% in gilts given the low dietary vitamin E

with no injected vitamin A and 80% in gilts given the high

dietary vitamin E with no injected A. Embryonic survival

was 100% in gilts injected with vitamin A and fed either the

low or high vitamin E diets. Brief and Chew (1985) reported

larger litter size and higher embryonic survival in gilts

receiving weekly injections of vitamin A (12,300 IU) and B-

carotene (33 mg) compared to gilts fed vitamin A and B-

carotene at the same levels. However, these gilts in the

study of Brief and Chew (1985) were depleted of vitamin A

and B-carotene for 5 weeks before the start of the study.

Coffey and Britt (1993) observed on average .5 pig increase

in the number of pigs born alive and higher embryonic

survival in sows given i.m. injections of vitamin A

palmitate (50,000 IU) compared to sows given vehicle only

(corn oil) on day of weaning, mating and 7 d postbreeding.

These sows were also supplemented with 11,000 IU of vitamin

A acetate per kg of diet.

Serum concentrations of a-tocopherol in gilts fed diets








51

supplemented with two levels of vitamin E with and without

injected vitamin A are reported in Tables 4-3 and 4-4.

Initial a-tocopherol serum concentrations in gilts were

similar (d -7). When dietary levels were increased from 25

to 500 IU of DL-a-tocopheryl acetate per kg of diet, serum

a-tocopherol concentrations increased (P < .01) by d 0 and

were maintained throughout the duration of the study (Table

4-4). This finding is in agreement with other studies that

have shown that dietary vitamin E compounds are effective in

increasing serum tocopherol, and that serum concentration

increased with increasing dietary vitamin E (Jensen et al.,

1988; Asghar et al., 1991; Mahan, 1991 and Anderson et al.,

In Press).

Serum a-tocopherol was not affected (P > .1) by

injecting gilts with vitamin A (retinyl palmitate) except on

d 7 (Table 4-4). On d 7, the gilts fed the low vitamin E

diet had similar (P > .1) serum tocopherol concentrations

regardless of vitamin A treatment, whereas gilts fed the

high vitamin E diet had higher (P < .08) serum tocopherol

concentrations when injected with vitamin A than gilts not

injected with vitamin A (Table 4-3).

Serum concentrations of retinol due to treatment are

shown in Tables 4-5 and 4-6. In general, there was no

consistent effect on serum retinol due to dietary vitamin E

supplementation level or injection of vitamin A. However, a

difference in serum retinol (P < .08) was observed on d 0 in











Table 4-3. Mean serum a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin
E and injected with vitamin A


Vit. E, IU/kg and vit. A inj.a"'
Sampling 25/No 25/Yes 500/No 500/Yes SE
day
----------------- g/ml---------------------

-7 .8 .9 1.1 .6 .17
0 1.2 1.2 3.6 3.6 .18
7 1.1 1.1 3.4 4.0 .15
24 1.3 1.3 3.8 3.6 .15

Note: Each mean is based on 7 or 8 observations.

aThree injections of 350,000 IU each.

bEffect of vitamin E (P < .01), d 0, 7, 24.


CE x A effect (P < .08), d 7.









53

Table 4-4. Main mean serum a-tocopherol concentrations
in gestating gilts given dietary additions of
vitamin E and injected with vitamin A


Vit. E. IU/kgq Vit. A inj.b.c
Sampling 25 500 No Yes SE
day
-------------------g/ml---------------------

-7 .8 .9 .9 .8 .12
0 1.2 3.6 2.4 2.4 .13
7 1.1 3.7 2.3 2.6 .11
24 1.3 3.7 2.5 2.5 .11


Note: Each mean is based on 14, 15 or 16

aEffect of vitamin E (P < .01), d 0, 7 an

bThree injections of 350,000 IU each.

cEffect of vitamin A (P < .06), d 7 only.


observations.

d 24.









54

Table 4-5. Mean serum retinol concentrations in gestating
gilts given dietary additions of vitamin E and
injected with vitamin A


Vit. E. IU/kq" and vit. A inij.b.
Sampling 25/No 25/Yes 500/No 500/Yes SE
day
-------------------g/ml-----------------
-7 .51 .55 .52 .51 .03
0 .44 .49 .51 .59 .03
7 .40 .43 .47 .42 .03
24 .46 .47 .48 .45 .03

Note: Each mean is based on 7 or 8 observations.

aEffect of vitamin E (P < .03), d 0 only.

bThree injections of 350,000 IU each.

cEffect of vitamin A (P < .08), d 0 only.









55

Table 4-6. Main mean serum retinol concentrations in
gestating gilts given dietary additions of vitamin
E and injected with vitamin A


Vit. E, IU/kqg Vit. A inj.b.c
Sampling 25 500 No Yes SE
day
------------------p g/ml------------------

-7 .53 .52 .52 .53 .02
0 .46 .55 .47 .54 .02
7 .41 .44 .43 .42 .02
24 .46 .47 .47 .46 .01

Note: Each mean is based on 14, 15 or 16 observations.

"Vitamin E effect (P < .03) d 0 only.

bThree injections of 350,000 IU each.

cVitamin A effect (P < .08) d 0 only.








56

that serum retinol concentration was highest in gilts fed

the high vitamin E and injected with vitamin A, and lowest

in gilts fed low vitamin E without injected A. The

difference noted was very small and within normal values for

serum retinol concentrations usually found in the pig. Our

findings agree with Weaver et al. (1989) in that plasma

vitamin A concentration was not affected by dietary level of

vitamin E. Serum retinol concentrations in the chick and

rat have also been reported not to be affected by dietary

supplementation of vitamin E in studies done by Abawi and

Sullivan, (1989) and Blakely et al. (1991), respectively.

Mooney (1992) injected gilts with vitamin A palmitate

(ranging from 53,200 to 106,400 IU given once weekly), or B-

carotene (106.4 to 425.6 mg) and observed no difference in

plasma concentrations of either retinol or B-carotene. In

contrast, Brief and Chew (1985) noted increased plasma

vitamin A concentration upon injecting vitamin A, however,

gilts used in their research were depleted of vitamin A

prior to the study and they also received injected B-

carotene. Serum retinol may have been elevated with

injection of vitamin A early in our study but may have been

missed since blood samples were taken 7 d after injection.

Tissue a-tocopherol concentration in gestating gilts

increased (P < .01) as dietary supplementation of vitamin E

increased (Table 4-7) in all tissues except adipose. This

finding is in agreement with others who have observed











Table 4-7. Mean tissue a-tocopherol concentrations of
gestating gilts given dietary additions of vitamin E
and injected with vitamin A


Vit. E, IU/kg and vit. A inj.b
Tissuea 25/No 25/Yes 500/No 500/Yes SE
--g/---- -g--..-----------------g/g_

Liver 4 4 24 23 1.6
Back fat 6 8 9 9 1.5
Leaf fat 9 10 12 12 2.2
Semimembranosus 2 3 4 4 .3
Rhomboideus 3 3 7 7 .6
Endometriumd 2 1 4 5 .3
Embryo .4 .4 .8 .7 .05
Oviduct 1 1 4 3 .3
Uterus 1 1 4 4 .2
Ovary 20 19 104 90 5.6

Note: Each mean is based on 7 or 8 observations.

aEffect of vitamin E (P < .01) for all tissues except
back fat and leaf fat.

bThree injections of 350,000 IU each.

CWet tissue basis.

dEffect of vitamin A (P < .08); E x A (P < .04).








58

similar responses in the pig (Jensen et al., 1988; Asghar et

al., 1991; Mahan, 1991). Average tocopherol concentration

increased by a factor of 2 in embryos upon high dietary

supplementation indicating that tocopherol is transferred

from the dam to the developing embryo. Vitamin A injections

had no effect (P > .1) on tissue a-tocopherol concentrations

except in the endometrium where there was a vitamin E x

vitamin A interaction (P < .04). In the endometrium gilts

fed low vitamin E and injected with vitamin A had slightly

lowered tocopherol concentration, while gilts fed the high

vitamin E and injected with vitamin A had increased

tocopherol concentration over that of the non injected gilts

(Table 4-7). Vitamin A injections appear to have no effect

on the transfer of tocopherol into the developing embryos as

tocopherol concentration in the embryos was not influenced

by vitamin A injection (P > .1).

Injecting vitamin A had no effect (P > .1) on retinol

concentration in any of the tissues studied including the

liver. Retinol concentrations in tissues other than liver,

however, were very small or nonexistent. Average

concentration in the liver was 386 gg/g. Mooney (1992)

found no differences in concentration of retinol in uterine

flushings in gilts that were injected with varying levels of

vitamin A; liver retinol was not determined.

Among the tissues sampled, the highest average

concentration of a-tocopherol upon supplementation of high








59

level of vitamin E was found in the ovary, followed by

liver, adipose, rhomboideus and endometrium, respectively

(Table 4-8). Three other tissues followed (semimembranosus,

oviduct, uterus) that had similar average concentrations and

embryo tissue had the lowest a-tocopherol concentration

(units per wet tissue basis).

In conclusion, there was no consistent evidence found

in this study that injecting a relatively large amount of

vitamin A (3 injections of 350,000 IU) just before, during

and shortly after breeding, significantly improved

reproductive performance, or interfered with serum or tissue

concentrations of a-tocopherol in gestating gilts fed diets

supplemented with 25 or 500 IU of vitamin E/kg of diet.

However, tocopherol concentration was increased further in

the endometrial tissue when vitamin A was given along with

high dietary vitamin E. No evidence was found that

injections of vitamin A interfered with the transfer of a-

tocopherol to the developing embryo. Likewise, serum

retinol concentrations were not affected by treatment.


Summary


A 2 x 2 factorial experiment was conducted to determine

the effects of injecting vitamin A and feeding vitamin E on

reproductive performance and on blood serum and tissue

concentrations of tocopherol during early gestation of

gilts. Thirty-two crossbred gilts were fed corn soybean-









60

Table 4-8. Main mean tissue a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin E
and injected with vitamin A


Vit. E, IU/kg Vit. A inj.b
Tissues' 25 500 No Yes SE
---------------- g/g-----------------

Liver 4 24 14 14 1.1
Back fat 7 9 7 9 1.0
Leaf fat 9 12 10 11 1.5
Semimembranosus 3 4 3 3 .2
Rhomboideus 3 7 5 5 .4
Endometriumd 1.4 4.6 2.8 3.3 .18
Embryo .4 .8 .6 .6 .03
Oviduct 1 4 3 2 .2
Uterus 1 4 3 3 .1
Ovary 19 97 61 55 3.8

Note: Each mean is based on 14, 15 or 16 observations.

aEffect of vitamin E (P < .01) for all tissues except
back fat and leaf fat.

bThree injections of 350,000 IU each.

Wet tissue basis.

dEffect of vitamin A (P < .08).








61

meal based diets supplemented with DL-a-tocopheryl acetate

to provide either 25 or 500 IU of added vitamin E/kg of

diet. Gilts were fed daily 1.9 kg/h beginning d -7

prebreeding through d 25 of gestation. Half of the gilts

were injected (i.m.) with 350,000 IU of retinol palmitate at

d -7 prebreeding, breeding (d 0) and d 7 postbreeding; the

other half were injected with vehicle only. All gilts were

double mated during their second or third estrus. Blood

samples were collected on d -7, 0, 7 and 24 of gestation.

Gilts were slaughtered on d 25 of gestation following

accepted slaughter procedures. Twenty-nine gilts conceived.

The number of corpora lutea and embryos was not affected (P

> .1) by treatment. Serum tocopherol concentrations

increased with 500 IU of vitamin E by d 0 and were stable

through d 24 of gestation (P < .01). Vitamin A injections

had no effect (P > .1) on serum tocopherol concentrations

except on d 7 when a small increase (P < .06) was noted.

High dietary vitamin E increased tocopherol concentration (P

< .01) in all tissues examined except fat. A vitamin E x

vitamin A interaction (P < .04) was noted in endometrium

tissue. Low dietary vitamin E and injections of vitamin A

slightly lowered tocopherol concentration, while high

vitamin E and vitamin A injections increased tocopherol

concentration in the endometrium. Increasing dietary

vitamin E increased serum and tissue tocopherol

concentrations. Vitamin A injections had little or no









62

effect on these concentrations during early gestation of

gilts.














CHAPTER 5
GENERAL CONCLUSIONS


Three experiments were conducted, one to determine the

bioavailability of four forms of vitamin E compounds, and

two to assess the effect of high levels of vitamin A on the

vitamin E status of growing finishing pigs or gestating

gilts.

In experiment 1 the biopotency of four forms of vitamin

E were determined. Generally, the acetate forms resulted in

greater serum and tissue concentrations of vitamin E (a-

tocopherol) than the alcohol forms, due to the greater

stability of the acetate forms that was noted in mixed feed.

Serum tocopherol increased rather rapidly when the four

compounds were fed. Dietary supplementation of D-a-

tocopheryl acetate resulted in the highest serum tocopherol

throughout the study, compared to concentrations obtained

for pigs fed the other compounds indicating a greater

biopotency (IU/mg) for swine than determined by the

traditional rat bioassay. A similar trend was observed with

tissue (liver, back fat, leaf fat, and muscle) tocopherol

concentrations as with serum concentrations, with the liver

having the highest concentration. In general, all forms

would probably be suitable dietary supplemental sources if








64

the stability of the alcohols were improved. Encapsulating

the alcohol forms to protect them from destruction would

increase their suitability for use in mixed feed.

Experiment 2 was conducted to evaluate the effect of

feeding excessive vitamin A on growth performance, and on

blood and tissue a-tocopherol (vitamin E) levels of growing-

finishing pigs. High dietary vitamin A (20,000 IU/kg of

diet) was found not to affect or have little affect on pig

performance, or on blood or tissue concentrations of a-

tocopherol. A threefold increase (P < .01) in serum

tocopherol occurred on all sampling days when dietary

supplementation increased from 15 to 150 IU/kg. Tissue

tocopherol also increased (P < .001) as dietary vitamin E

increased from 15 to 150 IU/kg. Tissue tocopherol

concentration increased (P < .001) by a factor of at least

two in all tissues evaluated. Liver retinol increased (P <

.001) eightfold with a tenfold increase in dietary vitamin

A. Even in the liver when vitamin A (retinol) concentration

was high, a-tocopherol concentration was not affected.

Thus, the form of vitamin A within the liver is not in a

form which can lead to oxidative destruction of a-tocopherol

or the concentrations encountered may not have been high

enough to affect a-tocopherol.

Experiment 3 evaluated the effect of giving a high

level of vitamin A via intramuscular injections on

reproductive performance, and on serum and tissue a-








65

tocopherol concentrations during early gestation of gilts.

High levels of vitamin A (350,000 IU per week) did not

affect reproductive performance, or serum or tissue

concentrations of a-tocopherol or retinol in this study. As

observed in the previous studies, increasing dietary levels

of vitamin E increased blood serum and tissue tocopherol

concentrations including reproductive tissues. Alpha-

tocopherol concentration also increased in the embryo when

dietary vitamin E was increased. The increased

concentration of a-tocopherol indicates a transfer of

tocopherol from the dam to the developing embryo.

Fluctuations of retinol concentrations in serum may have

been missed due to weekly sampling of blood. Blood sampling

on d 2, 3, or on a more frequent routine after vitamin A

injections would provide more answers.

Further research would be desirable to determine

vitamin E bioavailability other than by oral administration.

Fecal sample tocopherol analysis may provide more

information from which better conclusions might be drawn

concerning digestion and absorption. Studies using larger

numbers of gilts and/or sows is recommended to determine if

vitamin A injection alone or in combination with high

vitamin E, either orally or by injection, would enhance

reproductive performance.















REFERENCES


Abawi, F. G., and T. W. Sullivan. 1989. Interaction of
vitamins A, D3, E, and K in the diet of broiler chicks.
Poult. Sci. 68:1490.

Abawi, F. G., T. W. Sullivan, and S. E. Scheideler. 1985.
Interaction of dietary fat with levels of vitamin A and
E in broiler chicks. Poult. Sci. 64:1192.

Adams, C. R. 1978. Vitamin product forms for animal
feeds. In: Vitamin Nutrition Update, Seminar Series
2. Roche Publ. Nutley, NJ.

Ames, S. R. 1979. Biopotencies in rats of several forms
of alpha-tocopherol. J. Nutr. 109:2198.

Anderson, L. E.,Sr., R. O. Myer, J. H. Brendemuhl, and L.
R. McDowell. Bioavailability of various vitamin E
compounds for finishing swine. J. Anim. Sci. In press.

Arnrich, L., and V. A. Arthur. 1980. Interaction of fat-
soluble vitamins in hypervitaminoses. Ann. of N. Y.
Acad. Sci. 355:109.

Asghar, A., J. I. Gray, E. R. Miller, P. K. Ku, A. M.
Booren, and D. J. Buckley. 1991. Influence of
supranutritional vitamin E supplementation in the feed
on swine growth performance and deposition in different
tissues. J. Sci. Food Agric. 57:19.

Baker, H., G. J. Handelman, S. Short, L. J. Machlin, H.
N. Bhagavan, E. A. Dratz, and 0. Frank. 1986.
Comparison of plasma a and gamma tocopherol levels
following chronic oral administration of either all-
rac-a-tocopheryl acetate or RRR-a-tocopheryl acetate in
normal adult male subjects. Am. J. Clin. Nutr. 43:382.

Behrens, W. A., and R. Madere. 1991. Tissue discrimination
between dietary RRR-a- and all-rac-a-tocopherols in
rats. J. Nutr. 121:454.

Blakely, S. R., G. V. Mitchell, M. Y. Jenkins, E.
Grundel, and P. Whittaker. 1991. Canthaxanthin and











excess vitamin A alter a-tocopherol, carotenoid and
iron status in adult rats. J. Nutr. 121:1649.

Brandner, G. 1971. "Vitamin E." Bell Publishing Company,
New York.

Bratzler, J. W., J. K. Loosli, V. N. Krukousky, and L. A.
Maynard. 1950. Effect of the dietary level of
tocopherols on their metabolism in swine. J. Nutr.
42:59.

Brief, S., and B. P. Chew. 1985. Effects of vitamin E and B-
carotene on reproductive performance in gilts. J.
Anim. Sci. 60:998.

Burton, G. W., K. H. Cheeseman, T. Doba, K. U. Ingold, and
T. F. Slater. 1983. Biology of Vitamin E. Ciba
Foundation Symposium 101. Pitman, London.

Chew, B. P., T. S. Wong, J. J. Michael, F. E. Standaert, and
L. R. Heirman. 1991. Kinetic characteristics of B-
carotene uptake after an injection of B-carotene in
pigs. J. Anim. Sci. 69:4883.

Chung, Y. K., D. C. Mahan, and A. J. Lepine. 1992. Efficacy
of dietary d-a-tocopherol and dl-a-tocopheryl acetate
for weanling pigs. J. Anim. Sci. 70:2485.

Coelho, M. B. 1991. "Vitamin E in Animal Nutrition and
Management." BASF Corp. Parsippany, NJ.

Coffey, M. T., and J. H. Britt. 1993. Enhancement of sow
reproductive performance by B-carotene or vitamin A.
J. Anim. Sci. 71:1198.

Cunha, T. J. 1977. "Swine Feeding and Nutrition."
Academic Press, New York.

Diplock, A. T. 1985. "Fat-Soluble Vitamins." Technomic
Publishing Co. Inc., Lancaster, Pennsylvania.

Dove, C. R., and R. C. Ewan. 1991. Effect of trace
minerals on the stability of vitamin E in swine grower
diets. J. Anim. Sci. 69:1994.

Draper, H. H. 1980. Nutrient relationships. In: Machlin,
L. J. "Vitamin E: A Comprehensive Treatise." Marcel
Dekker, New York.

Erdman, J. W., Jr., C. L. Poor, and J. M. Dietz. 1988.
Factors affecting the bioavailability of vitamin A,
carotenoids, and vitamin E. Food Technol. 42:214.










Evans, H. M. 1962. The pioneer history of vitamin E.
Vitam. and Horm. 20:379.

Harris, P. L., and M. I. Ludwig. 1949a. Vitamin E
potency of a-tocopherol esters. J. Biol. Chem.
180:611.

Harris, P. L., and M. I. Ludwig. 1949b. Relative
vitamin E potency of natural and of synthetic a-
tocopherol. J. Biol. Chem. 170:1111.

Hatam, L. J., and H. J. Kayden. 1979. A high-performance
liquid chromatographic method for the determination of
tocopherol in plasma and cellular elements of the
blood. J. Lipid Res. 20:639.

Hidiroglou, N., N. Cave, A. S. Atwal, E. R. Farnworth, and
L. R. McDowell. 1992. Comparative vitamin E
requirements and metabolism in livestock. Ann. Rech.
Vet. 23:337.

Hidiroglou, N., and L. R. McDowell. 1987. Plasma and
tissue levels of vitamin E in sheep following
intramuscular administration in an oil carrier.
Internat. J. Vit. Nutr. Res. 57:261.

Hidiroglou, N., L. R. McDowell, and R. Pastrana. 1988.
Bioavailability of various vitamin E compounds in
sheep. Internat. J. Vit. Res. 58:189.

Hoppe, P. P., F. J. Schoner, and M. Frigg. 1992. Effects of
dietary retinol on hepatic retinol storage and on
plasma and tissue a-tocopherol in pigs. Internat. J.
Vit. Nutr. Res. 62:121.

Horwitt, M. K., W. H. Elliott, P. Kanjananggulpan, and C. D.
Fitch. 1984. Serum concentrations of a-tocopherol after
ingestion of various vitamin E preparations. Am. J.
Clin. Nutr. 40:240.

Howard, K. A., S. V. Radecki, E. R. Miller, A. J. Thulin,
and D. E. Ullrey. 1990. Relative bioavailability of
vitamin E of natural or synthetic origin in growing
pigs. Res. Rep. 502. Michigan State Univ. Agric.
Experiment Station, East Lansing, MI.

Jensen, M., J. Hakkarainen, A. Lindholm, and L. Jonsson.
1988. Vitamin E requirement of growing swine. J. Anim.
Sci. 66:3101.

Jensen, M., A. Lindholm, and J. Hakkarainen. 1990. The
vitamin E distribution in serum, liver, adipose and











muscle tissues in the pig during depletion and
repletion. Acta Vet. Scand. 31:129.

Kaneko, J. J. 1980. "Clinical Biochemistry of Domestic
Animals." (3rd Ed.). Academic Press, New York.

Koch-Weser, J. 1974. Medical intelligence. The New England
J. of Med. 29(5):233.

Kusin, J. A., V. Reddy and B. Sivakumar. 1974. Vitamin E
supplements and the absorption of a massive dose of
vitamin A. Am. J. Clin. Nutr. 27:774.

Machlin, L. J. 1980. "Vitamin E: A Comprehensive
Treatise." Marcel Dekker, New York.

Mahan, D. C. 1991. Assessment of the influence of dietary
vitamin E on sows and offspring in three parities:
reproductive performance, tissue tocopherol, and
effects on progeny. J. Anim. Sci 69:2904.

Mahan, D. C., and A. L. Moxon. 1980. Effect of dietary
selenium and injectable vitamin E-selenium for weanling
swine. Nutr. Rep. Int. 21:829.

Malm, A., E. F. Walker Jr., M. Homan, D. Kirtland, A. Aydin,
and W. G. Pond. 1976. Glutathione peroxidase and other
enzymes in serum of sow and their progeny fed vitamin
E adequate or deficient diets with added Se. Nutr. Rep.
Int. 14:185.

Mason, K. E. 1980. The First Two Decades of Vitamin E
History. In: Machlin, L. J., "Vitamin E: A
Comprehensive Treatise." Marcel Dekker, Inc., New
York.

McDowell, L. R. 1989. "Vitamins in Animal Nutrition."
Academic Press, Inc., San Diego, CA.

McDowell, L. R., J. A. Froseth, R. C. Piper, I. A. Dyer, and
G. H. Kroening. 1977. Tissue selenium and serum
tocopherol concentrations in selenium-vitamin E
deficient pigs fed pea (Pisam sativum). J. Anim. Sci.
45:1326.

McMurray, C. H., and W. J. Blanchflower. 1979a. Application
of a high-performance liquid chromatographic
fluorescence method for the rapid determination of a-
tocopherol in the plasma of cattle and pigs and its
comparison with direct fluorescence and high-
performance liquid chromatography-ultraviolet detection
methods. J. Chromatogr. 178:525.











McMurray, C. H., and W. J. Blanchflower. 1979b.
Determination of a-tocopherol in animal feedstuffs
using high-performance liquid chromatography with
spectrofluorescence detection. J. Chromatgr. 176:488.

Mooney, K. 1992. The effects of supplemental vitamin A
or B-carotene on reproduction and the localization
of B-carotene during gestation in gilts. M.S. Thesis.
Univ. of Florida, Gainesville.

Njeru, C.A., L. R. McDowell, N. S. Wilkinson, S. B. Linda,
S. N. Williams, and E. L. Lentz. 1992. Serum a-
tocopherol concentration in sheep after intramuscular
injection of DL-a-tocopherol. J. Anim. Sci. 70:2562.

NRC. 1988. "Nutrients Requirements of Swine." (9th Ed.).
National Academy Press, Washington, DC.

Pudelkiewicz, W. J., L. Webster, and L. D. Matterson. 1964.
Effects of high levels of dietary vitamin A acetate on
tissue tocopherol and some related observations. J.
Nutr. 84:113.

Raacke, I. D. 1983. Herbert McLean Evans: a biographical
sketch. J. Nutr. 113(5):929.

SAS. 1988. SAS User's Guide: Statistics." SAS Inst. Inc.,
Cary, NC.

Scott, M. L. 1969. Studies of vitamin E and related
factors in nutrition and metabolism. In: DeLuca,
H. F., and J. W. Suttie. "The Fat Soluble Vitamins."
The University of Wisconsin Press, Madison.

Sklan, D., and S. Donoghue. 1982. Vitamin E response to
high dietary vitamin A in the chick. J. Nutr. 112:759.

Tappel, A. L. 1962. Vitamin E as the biological lipid
antioxidant. Vitamins and Hormones. 20:493.

"The American Heritage Dictionary." 1982. (2nd Ed.).
Houghton Mifflin, Boston.

Thompson, J. 1993. Vitamin E injections improve pig
survival. Feedstuffs 65(21):13.

Ullrey, D. E. 1981. Vitamin E for swine. J. Anim. Sci.
53:1039.

Weaver, E. M., G. W. Libal, C. R. Hamilton, and I. S.
Parker. 1989. Relationship between dietary vitamin A
and E on performance and vitamin E status of the weaned
pig. J. Anim. Sci. 67(Suppl. 2):113. (Abstr.).














BIOGRAPHICAL SKETCH


Lee E. Anderson, Sr., was born December 1944 in

Pineland, South Carolina. He is married to Erma L. Anderson

and the father of five children. He graduated from

Middleton Senior High School in 1962. He graduated with an

Associate of Arts degree in 1964 from Gibbs Junior College.

He received a Bachelor of Science in Agriculture in 1969

from Florida A and M University. He received the Master of

Science in reproductive physiology from the University of

Florida in 1972. Since receiving the M.S. he has been

employed at Alcorn State University, Fort Valley State

University and Florida A and M University, in extension,

research and instructional programs. He is presently a

candidate for the Ph.D.








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.



Robert 0. Myer, Chairman
Associate Professor of Animal
Science

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.



Joel H. Brendemuhl'
Associate Professor of Animal
Science

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.



Ji'mmy G. cyeek
Professor of Agricultural
'Education and Communication

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 dre of Doctor of Philosophy.



J ph HU C nr
ofessor of Animal Science

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.



Lee R. McDowell
Professor of Animal Science








This dissertation was submitted to the Graduate Faculty
of the College of Agriculture and to the Graduate School and
was accepted as partial fulfillment of the requirements for
the degree of Doctor of Philosophy.


August, 1993


Dean, C Yege of Agric ture


Dean, Graduate School










































UNIVERSITY OF FLORIDA


3 1262 08556 9464




Full Text
VITAMIN E STATUS IN SWINE AS AFFECTED BY FORM OR LEVEL OF
DIETARY VITAMIN E AND/OR BY SUPPLEMENTATION OF VITAMIN A
By
LEE E. ANDERSON, SR.
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
1993
UNIVERSITY OF FLORIDA LIBRARIES

This dissertation is dedicated to my wife, Erma, and
children, Valerie, Lee Jr., Jenaya, Calvin, and Kelvin, and
to the memory of my parents, Florence Anderson, Myrtis
Stepherson and Leon Anderson, for their love, support and
encouragement.

ACKNOWLEDGMENTS
The author sincerely appreciates the efforts of all
members of his supervisory committee. Dr. Robert 0. Myer is
greatly appreciated for his understanding, guidance, advice
and companionship over the past several years. Sincere
thanks go to Dr. Joel Brendemuhl for his help and guidance
and his expertise and assistance during data collection.
The author is very appreciative of Dr. Lee McDowell's help
and advice, especially his assistance with acguiring the
vitamins. The advice, assistance and encouragement of Dr.
Jimmy Cheek and Dr. Joseph Conrad are deeply appreciated.
Thanks go to Dr. George Combs for his assistance. Without
the support of the above-mentioned individuals, this
dissertation would not have been possible.
Special gratitude is kindly extended to Dr. Jack Fry,
Dr. Roderick McDavis and Dr. Charles Kidd for their
encouragement and assistance during the course of this
program. Thanks are extended to Mr. Stephen Linda for the
valuable assistance he has given in the statistical analysis
of the data collected.
To his fellow graduate students, Maria Soler, Kim and
Kelly Sheppard, Dave Davis and Ken Mooney, the author
extends his thanks for their contributions and sacrifices in
iii

assisting him in this research. Special appreciation is
extended to Maria Soler and Kim Sheppard for their time and
involvement in trial 3.
Deep appreciation is extended to Alvin Boning for his
companionship and enthusiastic assistance in the laboratory.
Thanks go to Nancy Wilkinson for her assistance in the
laboratory. The author would like to take this opportunity
to thank Dane Bernis, Tom Crawford, Dennis Perry, Jack
Stokes, Dean Glicco and Rome Williams for their involvement
in the preparation of diets, care and management of the
experimental animals. Special appreciation is extended to
Leroy Washington, Larry Eubanks and Art Rogers for their
assistance in collecting tissue samples.
The author thanks Dr. Scot Williams and Hoffmann-La
Roche for providing the vitamins used in trials 2 and 3.
A special thanks is also extended to Mary Chambliss for
typing the manuscripts for review and publication.
IV

TABLE OF CONTENTS
Page
ACKNOWLEDGMENTS iii
LIST OF TABLES vi
ABSTRACT viii
CHAPTERS
1 INTRODUCTION 1
2 POTENCY OF VARIOUS VITAMIN E COMPOUNDS FOR
FINISHING SWINE 9
Introduction 9
Experimental Procedures 10
Results and Discussion 14
Summary 2 5
3 THE EFFECT OF EXCESSIVE DIETARY VITAMIN A ON
PERFORMANCE AND VITAMIN E STATUS IN SWINE FED
DIETS VARYING IN DIETARY VITAMIN E 27
Introduction 27
Experimental Procedures 28
Results and Discussion 32
Summary 4 2
4 EFFECT OF INJECTED VITAMIN A AND DIETARY
SUPPLEMENTATION OF VITAMIN E ON REPRODUCTIVE
PERFORMANCE AND TOCOPHEROL STATUS IN GESTATING
GILTS 44
Introduction 44
Experimental Procedures 45
Results and Discussion 48
Summary 59
5 GENERAL CONCLUSIONS 63
REFERENCES 66
BIOGRAPHICAL SKETCH 71
v

LIST OF TABLES
Table Page
2-1 Composition of finisher diet 11
2-2 Performance of finishing pigs fed diets containing
various vitamin E compounds 15
2-3 Vitamin E (a-tocopherol) concentrations in feed... 17
2-4 Adjusted serum vitamin E (tocopherol) concentrations
in finishing swine fed diets supplemented with
different vitamin E compounds 19
2-5 Relative biopotency of vitamin E compounds (%).... 22
2-6 Adjusted tissue a-tocopherol concentrations in
finishing swine fed diets supplemented with
different vitamin E compounds 23
3-1 Composition of diets (%) 29
3-2 Performance of growing-finishing swine fed diets
with different dietary levels of vitamin E
and vitamin A 33
3-3 Mean a-tocopherol concentrations in blood serum
due to dietary additions of vitamins E and A 34
3-4 Main means of serum a-tocopherol due to dietary
additions of vitamins E and A 35
3-5 Main means of serum retinol due to dietary additions
of vitamins E and A 38
3-6 Main means of tissue a-tocopherol concentrations due
to dietary additions of vitamins E and A 40
3-7 Main means of tissue retinol concentrations due to
dietary additions of vitamins E and A 41
4-1 Composition of diet fed to gestating gilts 46
vi

4-2 Mean reproductive response criteria of gestating
gilts given dietary additions of vitamin E and
injected with vitamin A 49
4-3 Mean serum a-tocopherol concentrations in gestating
gilts given dietary additions of vitamin E and
injected with vitamin A 52
4-4 Main mean serum a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin
E and injected with vitamin A 53
4-5 Mean serum retinol concentrations in gestating gilts
given dietary additions of vitamin E and injected
with vitamin A 54
4-6 Main mean serum retinol concentrations in gestating
gilts given dietary additions of vitamin E and
injected with vitamin A 55
4-7 Mean tissue a-tocopherol concentrations of gestating
gilts given dietary additions of vitamin E and
injected with vitamin A 57
4-8 Main mean tissue a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin E
and injected with vitamin A 60
vii

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
VITAMIN E STATUS IN SWINE AS AFFECTED BY FORM OR LEVEL OF
DIETARY VITAMIN E AND/OR BY SUPPLEMENTATION OF VITAMIN A
By
Lee E. Anderson, Sr.
August, 1993
Chairman: R. 0. Myer
Major Department: Animal Science
Experiment one used 40 finishing pigs (80 kg) to
determine the potency of vitamin E compounds. Pigs were
divided among five nutritionally adequate diets supplemented
with DL-a-tocopherol, DL-a-tocopheryl acetate, D-a-
tocopherol, D-a-tocophery1 acetate or no vitamin E. Blood
and tissue samples were collected. Vitamin E forms
increased (P < .05) serum a-tocopherol concentrations by d 2
of the feeding period. Serum tocopherol in pigs fed acetate
forms remained elevated through out the study; serum
concentrations declined (P < .01) in pigs fed alcohol forms.
D-a-tocophery1 acetate resulted in highest serum and tissue
tocopherol. The potency of D-acetate form was greater for
swine than that predicted from bioassays with the rat.
Experiment two evaluated excessive dietary vitamin A on
vitamin E status and performance of growing-finishing pigs.
viii

Eighty-four pigs were fed corn-soybean meal based diets
supplemented with DL-a-tocophery1 acetate to provide 0, 15
or 150 IU of vitamin E/kg and with retinyl acetate to
provide 2,000 or 20,000 IU of vitamin A/kg of diet. Serum
and tissue tocopherol increased (P < .01) as dietary levels
of vitamin E increased. The data indicated that 20,000 IU
of vitamin A/kg of feed did not affect performance or serum
and tissue tocopherol.
In experiment three, 32 gilts were used to determine
the effects of vitamins A and E on reproductive performance
and on serum and tissue concentrations of vitamin E during
early gestation. Treatments consisted of corn-soybean meal
based diets supplemented with DL-a-tocophery1 acetate to
provide either 25 or 500 IU of vitamin E/kg of diet,
beginning d -7 prebreeding through d 25 of gestation. Half
of the gilts were injected with 350,000 IU of vitamin A
(retinyl palmitate) at d -7, again at breeding (d 0), and at
d 7 postbreeding. Reproductive performance was not affected
by treatment. Serum tocopherol increased (P < .01) with 500
IU of vitamin E. High (500 IU/kg) dietary vitamin E
increased tocopherol level (P < .01) in all tissues except
adipose. High vitamin A (350,000 IU) via injections had no
consistent effect on reproductive performance or on serum or
tissue concentrations of a-tocopherol or retinol.
IX

CHAPTER 1
INTRODUCTION
Vitamin E was discovered in 1922 as a missing, needed
dietary factor (Brandner, 1971; Ullrey, 1981; Raacke, 1983;
McDowell, 1989). Vitamin E was isolated as alpha-
tocopherol. The name tocopherol means to bring forth
offspring (McDowell, 1989). George M. Calhoun, a professor
of Greek at the University of California, Berkeley, named
the new vitamin tocopherol in 1936 (tocos for childbirth,
phero to confer, and ol for alcohol) (Evans, 1962; Ullrey,
1981; Raacke, 1983).
It was recognized in 1920 that reproductive failure
occurred in rats consuming diets thought to be nutritionally
adequate. An unknown dietary factor, then called X and
later determined to be vitamin E, was deficient, which
resulted in fetal death and embryo resorption in the
laboratory rat (Evans, 1962; Mason, 1980; Diplock, 1985;
McDowell, 1989). Estrus and mating were normal, but fetuses
died and were resorbed unless the diet was supplemented with
small amounts of wheat germ, dried alfalfa leaves, or fresh
lettuce, which contained the deficient vitamin E (Evans,
1962; Mason, 1980; McDowell, 1989). Degeneration of the
germinal epithelium in male rats was prevented by
1

2
supplements of fresh lettuce (Mason, 1980). Other animal
species (cattle, sheep, mink, and chickens) were able to
reproduce without dietary vitamin E, but in each case their
offspring died prematurely (Brandner, 1971).
Vitamin E became known as the fertility vitamin.
Many studies were done to determine if vitamin E affected
reproduction in humans. In most cases vitamin E had little
or no effect (McDowell, 1989).
Vitamin E deficiency in swine results in reduced
reproductive efficiency, locomotor incoordination, muscular
and hepatic necrosis, fibrinoid degeneration of blood vessel
walls and muscular dystrophy (McDowell, 1977).
Vitamin E is a hydrophobic, peroxyl radical-trapping,
chain-breaking antioxidant found in the lipid fraction of
living organisms. Its principal function is to protect the
lipid material of an organism from oxidation (Machlin, 1980;
Burton et al., 1983; Diplock, 1985; McDowell, 1989; Coelho,
1991). Lipid peroxidation of membranes of cells and
cellular constituents can be very damaging. Damage may be
as simple as breaking a membrane and allowing leakage of
contents, or as complex as breaking a membrane containing
destructive enzyme systems. Hemolysis of red blood cells is
an example of relatively simple membrane breakage. Membrane
damage to lysosomes can be particularly devastating.
Lysosomes are sometimes called the "suicide bags" of the
cell, and when their membranes are broken they release

3
enzymes that hydrolyze tissue constituents and magnify
tissue damage (Tappel, 1962). Damage to the membrane of
such other cellular components as mitochondria and
microsomes, which contain 25 and 40 % unsaturated lipid,
respectively, have profound effects. In both microsomes and
mitochondria, vitamin E is the only known lipid antioxidant
(Tappel, 1962).
Selenium (Se) is a trace mineral that is known to spare
some of the requirement for vitamin E. Selenium is a
component of the enzyme glutathione peroxidase, which is a
selenoprotein containing four atoms of selenium per molecule
of protein (Scott, 1969; Draper, 1980). Glutathione is a
hydrogen donor. Vitamin E functions as a fat soluble
antioxidant, and selenium functions as a water soluble
antioxidant (Cunha, 1977). Vitamin E is the first line of
defense against peroxidation of fats in cells. If peroxides
are formed, selenium through the enzyme glutathione
peroxidase destroys the peroxides before tissue damage can
occur. Thus, selenium is considered the second line of
defense (Diplock, 1985; McDowell, 1989) and as a result,
both selenium and vitamin E are capable of preventing some
of the same nutritional diseases (McDowell, 1989). Vitamin
E can also reduce the selenium requirement by inhibiting
production of peroxides.
Pigs exhibiting clinical vitamin E and Se deficiency
signs have a pale, white discoloration of the skeletal

4
muscle, an enlarged, friable heart, associated with
hydropericardium, and sometimes intestinal edema and
hepatosis dietética (Mahan and Moxon, 1980) .
There are many factors that affect the bioavailability
of vitamin E. These include the form of vitamin E compound,
potency of compound, stability, absorption, other fat
soluble vitamins (e.g., retinol), mineral interactions, and
unsaturated fat. Bioavailability is defined as the
percentage of a drug or nutrient (in this case, vitamin E)
that enters the systemic circulation after administration
and the rate of entry into the general circulation for
distribution throughout the body as well as tissue
accumulation (Koch-Weser, 1974; "The American Heritage
Dictionary", 1982).
Eight forms of vitamin E are known to occur in nature,
four of which are referred to as tocopherols and four as
tocotrienols. They have been given Greek letter names to
distinguish them from one another (Diplock, 1985; NRC,
1988). The compounds differ in the placement of methyl
groups on the ring and the degree of saturation in the side
chain (McDowell, 1989).
Alpha-tocopherol is a yellow oil, soluble in certain
organic solvents. It is common practice to assay only this
isomer rather than all eight compounds because a-tocopherol
is the most biologically active, naturally occurring vitamin
E source (Ullrey, 1981) .

5
DL-a-tocopherol has a potency of 1.1 IU/mg and its
acetate (DL-a-tocophery1 acetate) has a potency of 1 IU/mg
as determined by bioassays with rats. Activity of naturally
occurring a-tocopherol, D-a-tocopherol (also called RRR-
tocopherol) is 1.49 IU/mg and of its acetate, 1.36 IU/mg.
D-a-tocopherol is the most biologically active form (IU per
unit of weight; NRC, 1988).
Loss of vitamin E potency occurs in mixed feed from a
number of factors. The naturally occurring tocopherols have
relatively poor stability during processing, grinding,
pelleting, and storing at high temperatures or under moist
conditions. Vitamin E will also readily interact with other
ingredients in feed formulations (Adams, 1978; NRC, 1988).
More pigs are being raised in confinement without
access to pasture, which is an excellent source of vitamin
E. Heating and pelleting feed grains lower their vitamin E
values. The use of high moisture grain increases the need
for vitamin E supplementation due to the destruction of the
vitamin. Feeds formulated with fats containing high
quantities of unsaturated fatty acids are susceptible to
rancidity, which destroys vitamin E (Cunha, 1977). Malm et
al. (1976) reported that diets high in polyunsaturated fatty
acids increased vitamin E requirement and that pigs fed a
polyunsaturated fatty acid, low vitamin E diet throughout
the postweaning period resulted in some degree of red blood
cell destruction.

6
The alcohol form, a-tocopherol, is easily destroyed by
oxidation. Oxidative destruction of a-tocopherol is
accelerated by heat, light, moisture, unsaturated fats,
sulfates, nitrates and molds, and in diets containing
increased levels of copper, iron, zinc and manganese
(Ullrey, 1981; McDowell, 1989; Dove and Ewan, 1991; Mahan,
1991; Thompson, 1993). A more stable source of vitamin E is
a-tocopheryl acetate. Alpha-tocophery1 acetate is
chemically synthesized by esterification of a-tocopherol
with acetic acid. DL-a-tocophery1 acetate is the
international standard for vitamin E activity.
Vitamin E is fat soluble and as such its absorption is
associated with that of lipids. Vitamin E is absorbed in
the alcohol form. Vitamin E acetate is hydrolyzed to the
alcohol form in the small intestine prior to absorption.
Droplets of triglycerides are degraded by lipase and bile
into monoglycerides and free fatty acids, which form into
micelles. Micelles contain the lipid components including
the fat soluble vitamins. Vitamins are absorbed with the
fatty acids and monoglycerides. Triglycerides are re-formed
in the intestinal cell and packaged into chylomicrons.
Chylomicrons are absorbed into the lacteal ducts and carried
into the lymphatic system until they enter the general
circulation and are distributed to various tissues.
Factors interfering with digestion and absorption of lipid
affect the bioavailability of vitamin E.

7
Competition for absorption sites in the small
intestines among the fat soluble vitamins may affect
bioavailability of vitamin E. Vitamin A (retinol) may
interfere with both absorption and blood concentrations of
vitamin E. This has been demonstrated in chicks (Sklan and
Donoghue, 1982; Abawi and Sullivan, 1989) and rats (Blakely
et al., 1991). This effect appears to be due to increased
oxidation of vitamin E prior to the digesta reaching the
duodenum. This would result in vitamin E concentration
being lower at the major absorption sites in the upper small
intestine (Sklan and Donoghue, 1982). In this case, vitamin
E is oxidized at the expense of vitamin A. Erdman et al.
(1988) reported that vitamin E may protect vitamin A from
oxidation in the gastrointestinal tract and within cell
membranes. Reports that vitamin A toxicity in chicks has
been completely reversed with high dietary vitamin E
supplementation (Arnrich and Arthur, 1980) also indicate an
additional loss of vitamin E resulting in an increased need.
Young children, who were vitamin A deficient, absorbed more
vitamin A when given high supplemental levels of vitamin E
(Kusin et al., 1974) indicating that vitamin A may also
affect the availability of vitamin E. However, there is
very little or no research regarding the influence of
vitamin A on the vitamin E status of swine.
Recently there has been an interest in increasing
supplemental vitamin A levels via i.m. injections in

8
gestating gilts and sows. Extra vitamin A given just
before, during, and shortly after breeding has been reported
to improve reproductive performance in breeding swine (Brief
and Chew, 1985; Coffey and Britt, 1993). The elevation of
maternal plasma vitamin A is believed to improve embryonic
survival (NRC, 1988). The elevated vitamin A may also
affect bioavailability of vitamin E and/or its requirement.
Selection for increased growth rate and reproductive
performance increases dietary vitamin E requirements. In
addition, confinement rearing and feeding cereal-soybean
meal diets that vary considerably in vitamin E content, make
it important to insure that adequate levels of nutrients are
included in the diet. Fortification of diets adequately
supplemented with vitamins is extremely important in
optimizing performance under current production conditions.
More research is necessary to ascertain the significant
aspects of vitamin E and its enhancement or impediment on
performance under conventional swine production systems.
Therefore the focus of this manuscript is on the
bioavailability of vitamin E as affected by the type of
vitamin E compound fed, and the influence of vitamin A
supplementation on vitamin E status of growing-finishing
pigs and during early gestation of gilts.

CHAPTER 2
POTENCY OF VARIOUS VITAMIN E COMPOUNDS FOR FINISHING SWINE
Introduction
Vitamin E is an essential nutrient for normal growth,
health and reproduction in swine. Vitamin E requirement for
swine ranges between 10 and 22 IU/kg of diet (NRC, 1988).
Swine diets consisting mainly of corn and soybean meal
usually do not contain adequate amounts of vitamin E needed
to meet the pig's requirement (NRC, 1988). In addition, the
stability of all naturally occurring vitamin E forms are
very poor in mixed feed (Ullrey, 1981; Dove and Ewan, 1991;
Hidiroglou et al., 1992). Therefore, supplementation of
swine diets with a readily available form of vitamin E
assures that swine will receive the correct amount for
optimum performance.
Eight forms of vitamin E occur in nature (4
tocopherols, 4 tocotrienols). D-a-tocopherol has the
greatest biological activity (highest IU per unit weight;
NRC, 1988) but acetate and succinate forms are more stable
(Erdman et al., 1988). This experiment evaluated the
relative biopotencies of four forms of vitamin E (DL-a-
tocopheryl acetate, D-a-tocophery1 acetate, DL-a-tocopherol,
and D-a-tocopherol) when supplemented in the diet of
9

10
finishing swine. The concentration of a-tocopherol in blood
serum and tissue was used as an indicator of potency.
Experimental Procedures
Forty crossbred finishing pigs, 20 barrows and 20
gilts, with an average initial weight of 80 kg were randomly
assigned by sex to individual pens. Treatments were
randomly assigned to the pens such that each treatment
consisted of 8 pigs (4 barrows, 4 gilts). Treatments
consisted of the following supplemented vitamin E forms: DL-
a-tocopherol, DL-a-tocopheryl acetate, D-a-tocopherol, and
D-a-tocopheryl acetate. A negative control, which received
no supplemental vitamin E, was also included to give a fifth
treatment. Vitamin E forms used were pure forms supplied in
an unprotected oil solution (Sigma Chemical Co., St. Louis,
MO). Supplemental vitamin E was added to the diets such
that pigs consuming 3.2 kg of feed would received 200 IU per
day. Pigs were fed a corn-soybean meal finishing diet
formulated with a modified vitamin premix (exclusive of
vitamin E) and .1 ppm of added selenium. Diets were
otherwise formulated following NRC (1988) guidelines.
Composition of the corn-soybean meal basal diet is given in
Table 2-1. The pigs were fed the finisher diet for 28 days.
Prior to the start of this trial all pigs were fed a diet
that contained 22 IU of vitamin E/kg of diet. Feed and
water were available ad libitum throughout the experiment.

11
Table 2-1. Composition of finisher diet
Ingredient
%, as fed
Ground corn
Soybean meal (48%)
Dynafos
Ground limestone
Salt
Trace mineral3
82.05
15.00
1.70
0.80
0.25
0.10
0.05
0.05
Vitamin mix'
Se premix1
“Provided 200 ppm zinc, 100 ppm iron, 55 ppm manganese,
11 ppm copper, and 1.5 ppm iodine.
bProvided 2.2 mg riboflavin, 11 mg niacin, 9 mg
pantothenic acid, 150 mg choline chloride, 11 ug vitamin
B,2, 1.5 mg vitamin K, 2750 IU vitamin A, and 440 IU
vitamin D3 per kg of diet.
“Provided 0.1 ppm selenium.

12
Pigs were housed in an open-sided building with solid
concrete floors. Individual pig weights and feed
consumption were recorded biweekly. The trial was carried
out in the spring (April-May) of the year. Pigs were
managed according to acceptable management practices
throughout the experiment. Protocol for animal care had
been approved by the University Animal Use Committee.
Blood samples were collected by jugular vein puncture
from each pig on d 0, 1, 2, 7, 14, 21, and 28 of the feeding
period. Blood samples were centrifuged after collection,
and serum was frozen and stored at -20°C until analyzed.
Feed samples were taken from the feeders on d 0, 5, 14 and
21, frozen and stored at -20°C until analyzed for a-
tocopherol concentration. On d 29, the 20 barrows were
slaughtered, using accepted slaughter procedures, at the
University of Florida meats laboratory and tissue samples
collected. Tissue samples included liver, muscle
(rhomboideus and semimembranosus), back fat (10th rib area)
and leaf fat. Tissue samples were frozen following
collection and stored at -20°C until analyzed.
Procedures used for the extraction and determination of
a-tocopherol in blood serum were as previously described
(Njeru et al., 1992). Procedures were similar to those used
by McMurray and Blanchflower (1979a,b) except in our study,
propanol was used in the serum extraction instead of
ethanol. Most of the vitamin E activity in serum and tissue

13
was assumed to be a-tocopherol (Ullrey, 1981). Extraction
of vitamin E from tissues and feed was done using a
procedure outlined by Chung et al. (1992). This procedure
was a modification of that of McMurray and Blanchflower
(1979b) and Hatam and Kayden (1979).
Alpha-tocopherol concentration was determined using 50
ul of the reconstituted sample (serum, tissue or feed)
injected onto a LiChrosorb SI 60 column (Hibar Fertigsaule
RT pre-packed column RT 250-4 E, Merck, Darmstadt, Germany)
250 mm x 4 mm I.D. and using a Perkin Elmer 550 terminal
(Perkin-Elmer Corp. Analytical Instruments, Norwalk, CT), a
Perkin Elmer ISS-100 auto sampler, and a Perkin Elmer Series
4 Liquid chromatograph pump. The mobile phase consisted of
900:99:1 HPLC grade iso-octane, tetrahydrofuran and acetic
acid. The detector was a Perkin Elmer LS-4 Fluorescence
Spectrometer with an excitation wavelength of 290 nm and an
emission wavelength of 320 nm. Data were collected by a
Perkin Elmer LCI-100 Laboratory Computing Integrator. Flow
rate was 1 ml/min. The retention time of a-tocopherol was
5.2 minutes. Alpha-tocopherol (Eastman Kodak Company,
Rochester, NY) was used as a standard, and sample peaks and
retention times were compared to those of the standards.
Standard concentration was calculated to give a peak of 250
or 500 ng. Alpha-tocopherol concentration of samples was
calculated by the external standard method. Spiked samples
were found to have a mean recovery rate of 97%.

14
Potencies of the various vitamin E compounds were
determined by comparing areas under the time curve (AUC)
within the serum, feed and tissue samples. Serum and tissue
concentrations were adjusted based on actual feed intake and
a-tocopherol levels in the feed. Alpha-tocopherol
concentrations reported were adjusted to a constant feed a-
tocopherol concentration (d 0 feed level). Serum and tissue
means were analyzed using the general linear model procedure
(SAS, 1988). Analysis of variance compared treatment
differences in serum and tissue tocopherol concentrations.
Analysis of covariance was also applied to the serum data
using baseline (d 0 serum tocopherol) data as a covariate.
Treatment means were compared using the least significant
difference multiple comparison procedure.
Results and Discussion
Growth rate of all pigs was good over the duration of
the 28 d study. Daily feed intake and feed-to-gain ratio
were not affected (P > .1) by supplementation of the vitamin
E sources (Table 2-2); however, a slight improvement in
growth rate (P < .09) was obtained in pigs fed the D-a-
tocopherol form compared to the negative control. Asghar et
al. (1991) reported improved growth rates in growing¬
finishing pigs fed dietary levels of DL-a-tocophery1 acetate
at 100 IU/kg of diet compared to pigs fed 10 IU/kg of diet.
In contrast, Chung et al. (1992) found no difference in

15
Table 2-2. Performance of finishing pigs fed diets containing
various vitamin E compounds
Item
Vitamin E source3
DL-a-TAC
D-a-TAC
DL-a-TOH
D-a-TOH
Neg.
control
No.
of pigs
8
8
8
8
8
ADG,
kg
1.05b
1.0b
1.09bc
1.18c
1.02b
ADF,
kg
3.87
3.5
3.66
3.79
3.4
F/G
3.78
3.61
3.37
3.28
3.55
aDL-a-TAC = DL-a-tocopheryl acetate; D-a-TAC = D-a-
tocopheryl acetate; DL-a-TOH = DL-a-tocopherol; D-a-TOH =
D-a-tocopherol.
bcMeans within the same row with a different superscript
differ significantly (P < .09).

16
growth performance due to vitamin E source (encapsulated D-
a-tocopherol or DL-a-tocopheryl acetate) or level (16, 48
and 96 IU/kg) in trials with young, starting swine.
Alpha-tocopherol analysis of the diets containing the
vitamin E forms are reported in Table 2-3. The vitamin E
forms were included in the diet so that pigs would consume
62 IU of added vitamin E/kg of feed (72 IU/kg total).
However, there was considerable variation in analyzed levels
among the dietary treatments. Also, there was some
variation in feed consumption among treatments. Therefore,
data reported were adjusted based on analysis of diets and
mean treatment group feed consumption (Table 2-3).
The indicator used to determine biopotency of the
vitamin E compounds was the concentration of a-tocopherol in
serum and selected tissues. Bratzler et al. (1950) found
that plasma tocopherol concentration reflected level of
tocopherol ingested in trials with young pigs fed different
concentrations of tocopherol. They also observed increases
of tocopherol in various tissues. Other researchers
indicated that an animal's vitamin E status can be
determined by measuring a-tocopherol concentration in serum
and various tissue after oral administration: Baker et al.
(1986) with humans, Hidiroglou and McDowell (1987) with
sheep, and Jensen et al. (1990) and Asghar et al. (1991)
with pigs. Numerous studies have shown that dietary vitamin
E compounds are effective in elevating blood tocopherol

17
Table 2-3. Vitamin E (a-tocopherol) concentrations in feed
Treatment
(vitamin E source)
Sampling daya
0 5 14 21
IU/kg
DL-a-tocopheryl acetate
94 (72)
70(54)
64(49)
56(43)
D-a-tocopheryl acetate
113 (72)
104(66)
74(47)
72(46)
DL-a-tocopherol
94(72)
24(17)
21(16)
13(10)
D-a-tocopherol
83 (72)
22(19)
19(16)
15(13)
Neg. control
10
5
5
5
aDay samples were taken after start of trial. Samples
were taken directly from feeder then frozen until analyzed.
Numbers in parenthesis represent adjusted levels - adjusted
to a constant IU/kg extrapolated from d 0 levels.

18
concentration, and also that blood tocopherol concentration
increased with increasing dietary level of vitamin E
(Hidiroglou et al., 1988; Jensen et al., 1988; Behrens and
Madere, 1991; Asghar et al., 1991; Chung et al., 1992).
As expected, mean serum concentrations of tocopherol at
d 0 (baseline) were similar across all treatments (Table 2-
4). All vitamin E compounds fed in this experiment
increased (P < .01) serum tocopherol concentration. The
increase in serum tocopherol concentration was rapid. The
increase started on d 1, grew further by d 2 (P < .01), and
plateaued by d 7. Horwitt et al. (1984) noted in a study
with humans that serum a-tocopherol concentrations were
increased at 8 to 24 hr after ingestion of various vitamin E
forms. Howard et al. (1990) with pigs weaned at 28 d,
depleted of vitamin E for the next 38 d, and then fed 30 IU
of supplemental vitamin E in the form of D-a-tocophery1
acetate or DL-a-tocophery1 acetate per kg of diet noted a
rapid increase in blood a-tocopherol. Jensen et al. (1990),
in a study with pigs, 49 d old, also observed a rapid
increase in serum tocopherol concentrations after feeding
supplemental DL-a-tocophery1 acetate. In both of the above
swine studies, the first blood samples were taken 7 d after
the start of the feeding trial.
Serum tocopherol concentrations of pigs fed both
acetate forms were maintained beyond d 7; however, levels
dropped steadily in pigs fed the alcohol forms and were

19
Table 2-4. Adjusted serum vitamin E (tocopherol)
concentrations in finishing swine fed diets
supplemented with different vitamin E compounds
Day
Vitamin
E source3
Neg.
control
DL-a-TAC
D-a-TAC
DL-a-TOH
D-a-TOH
/i 0
.8
. 8
. 8
.8
. 9
1
. 9bc
1.2b
1. lb
1.3b
. 8C
2
1.3C
1.8b
1.4C
1.6bc
. 8d
7
1.5C
2.2b
1.4C
1.4C
. 6d
14
1.4C
1.8b
1. lc
1.3C
. 4d
21
1.4C
1.8b
. 6d
. 9d
. 3C
28
1.4C
1.7b
. 5d
. 5d
. 4d
Note: Each mean is based on eight observations. Adjusted to
constant intake of 72 IU/kg diet based on d 0 feed
analyses (Table 2-3). Day 0 serum values were not
adjusted.
aDL-a-TAC = DL-a-tocophery1 acetate; D-a-TAC = D-a-
tocopheryl acetate; DL-a-TOH = DL-a-tocopherol; D-a-TOH =
D-a-tocopherol.
kdcMeans within the same row with a different superscript
differ (P < .01).

20
lower (P < .01) on d 21 and 28 than the acetate forms. This
drop was probably due to poor stability of the alcohol forms
in the feed (Table 2-3). Degradation of vitamin E occurs
through oxidation, and is accelerated by light, alkali, heat
and trace minerals (Ullrey, 1981; Erdman et al., 1988; Dove
and Ewan, 1991; Hidiroglou et al., 1992). In the absence of
oxygen, tocopherols are relatively heat, light, and alkali
stable (Ullrey, 1981). Stability of a-tocopherol is
increased by acylation of the compound (Ullrey, 1981).
Acetate forms of vitamin E were found to be guite stable in
feed in our study. Acetate forms of vitamin E have also
been noted to be stable compounds by other researchers
(Harris and Ludwig, 1949a,b; Ullrey, 1981; Dove and Ewan,
1991; Chung et al., 1992). In general, serum tocopherol
concentrations observed in the present study were similar to
those of other studies in which pigs were fed diets
containing similar levels of added vitamin E (Jensen et al.,
1988; Asghar et al., 1991).
While all vitamin E forms evaluated rapidly increased
serum tocopherol concentrations, there was some evidence of
a slight difference in the rate of this increase. Average
serum concentration in pigs fed the DL acetate form was not
increased (P > .05) until d 2, whereas serum concentrations
of pigs fed the other compounds were increased (P < .05) on
d 1. Horwitt et al. (1984) found that D-tocopherol raised
blood a-tocopherol concentrations faster than the D- or DL-

21
a-tocopherol acetate forms in research done with humans.
D-a-tocopheryl acetate resulted in higher serum
tocopherol levels than the DL- acetate form. Relative to
DL-a-tocopheryl acetate, the D- form of the same compound
had an average biopotency of 146% (IU basis; Table 2-5) or
199% (weight basis; 146 X 1.36). Howard et al. (1990)
determined a relative biopotency of 218% (weight basis) for
D-cr-tocopheryl acetate relative to the DL- form of the same
compound in trials with growing pigs. Thus it would appear
that the D- acetate form has a higher biopotency for swine
than that determined from the traditional rat fetal-
resorption bioassays. However, Ames (1979) presented
evidence that the commonly accepted conversion value of 1.36
may be too low in trials evaluating the relative biopotency
of several vitamin E compounds using the rat fetal-
resorption assay. Both alcohol forms in our study exhibited
similar biopotencies, and there was evidence during the
early portion of the study that these forms were slightly
more biopotent than DL-a-tocopherol acetate (Table 2-5).
Alpha-tocopherol in tissues in general followed a
similar pattern to that observed with serum (Table 2-6).
Overall, pigs fed any of the compounds had tissue tocopherol
concentrations higher than the negative control. Other
researchers have also found that adding vitamin E to the
diet increased tissue concentrations (Bratzler et al., 1950;
Jensen et al., 1990; Asghar et al., 1991).

22
Table 2-5. Relative biopotency of vitamin E compounds (%)
Compound
day
1
day
2
day
7
day
14
day
21
day
28
Avg.
D-a-TAC
154
146
151
151
141
132
146
DL-a-TOH
122
110
123
124
32
83
99
D-a-TOH
124
114
118
140
46
73
103
Note: Based on serum values. DL-a-tocopheryl acetate = 100;
IU basis.

Table 2-6. Adjusted tissue a-tocopherol concentrations in
finishing swine fed diets supplemented with different
vitamin E compounds
Vitamin E Source3
Tissue DL-a-TAC D-a-TAC DL-a-TOH D-a-TOH Neg. Control
Liver
i
i
i
! »
! ^
4.9b
- Mg/g
2.6C
2.7C
1.3d
Leaf fat
2.6
2.4
2.3
1.5
1.4
Back Fat
2.4bc
2.8b
1.9cd
. 9C
1.3dc
Rhomboideus
2.6b
2.9b
1.6cd
1.9°
1.4d
Semimembranosus
1.0cd
2.5b
1.3cd
1.4C
. 9d
Note: Each mean is based on four observations. Adjusted to
constant intake of 72 IU/kg diet based on d 0 analyses (Table 2-3).
Negative control values not adjusted.
aDL-a-TAC = DL-a-tocophery1 acetate; D-a-TAC = D-a-tocophery1 acetate;
DL-a-TOH = DL-a-tocopherol; D-a-TOH = D-a-tocopherol.
bcdcMeans within the same row with a different superscript differ
significantly (P < .02).

24
Generally, acetate vitamin E forms resulted in greater
tissue tocopherol concentrations than alcohol forms, and the
difference was significant (P < .02) in the liver and
rhomboideus. The greater tissue levels reflected the
greater stability of the acetate forms in the feed over the
duration of the study. Liver tocopherol averaged 4.6 /xg/g
for the acetate forms and 2.6 /xg/g for the alcohol forms.
Between the two acetate forms, D-a-tocopheryl acetate
resulted in the same tissue tocopherol concentrations as the
DL- form, with exception of the semimembranosus in which the
D- form resulted in higher concentrations (P < .02). Tissue
tocopherol concentrations were similar for the two alcohol
forms.
Liver had the highest tocopherol concentration of all
tissues evaluated. The liver could be an indicator of
dietary vitamin E status or reflect immediate status.
Jensen et al. (1990) indicated that serum and liver a-
tocopherol concentrations reflected the short term vitamin E
status of the pig and that muscle and fat tissue
concentrations reflected the pig's long-term vitamin E
status.
The concentration of tocopherol in all tissues other
than the liver was similar (P > .10) in pigs fed the two
acetate forms, with the exception of semimembranosus as
noted above. Jensen et al. (1988), in trials with growing
pigs, also noted that the liver had the highest

25
concentration of tocopherol, followed by adipose tissue and
skeletal muscle. Also, Asghar et al. (1991), feeding
growing pigs DL-a-tocopheryl acetate at 100 IU/kg of diet,
observed higher tocopherol concentrations in the liver,
followed by the heart, lung and kidney.
In conclusion, all vitamin E compounds evaluated almost
immediately (by d 1) began to increase serum a-tocopherol
concentrations upon ingestion. Tissue a-tocopherol
concentrations were reflective of serum concentrations.
Vitamin E acetate forms fed to finishing pigs resulted in
higher serum and tissue tocopherol concentrations than the
alcohol forms due to their greater stability in mixed feed.
Among the two acetate forms evaluated, the D- form had a
greater biopotency for swine than that determined by
traditional assays.
Summary
Relative biopotencies of four chemical forms of vitamin
E supplemented in diets of finishing swine for 28 d were
evaluated. Forty crossbred pigs (80 kg), individually
penned, were divided equally among five treatments.
Treatments consisted of corn-soybean meal based diets
supplemented with DL-a-tocopherol, DL-a-tocopheryl acetate,
D-a-tocopherol or D-a-tocophery1 acetate. A treatment
without vitamin E supplementation (negative control) served
as the fifth treatment. Each compound was supplemented at

26
62 IU/kg of diet. Blood samples were collected on d 0, 1,
2, 7, 14, 21, and 28. On d 29, half of the pigs were
slaughtered to obtain tissue samples. Feed samples, taken
from feeders, were collected on d 0, 5, 14, and 21. All
vitamin E forms fed increased (P < .05) serum a-tocopherol
concentration by d 2 and the concentration peaked by d 7.
Serum tocopherol concentrations in pigs fed either acetate
form remained elevated beyond d 7; serum concentrations
steadily declined and were lower (P < .01) on d 21 and 28 in
pigs fed either alcohol form in comparison to acetate-fed
pigs. The decrease was probably a reflection of reduced
stability of the alcohol forms in the feed; the acetate
forms were found to be stable in the feed over the 28 d
study. Dietary supplementation of D-a-tocopheryl acetate
resulted in the highest serum tocopherol concentrations
throughout the study, compared to concentrations obtained
for pigs fed the other compounds. A similar trend was
observed in tissue (liver, back fat, leaf fat,
semimembranosus, rhomboideus) tocopherol concentrations as
with serum concentrations, with the liver having the highest
concentration. The biopotency of D-a-tocopheryl acetate for
swine appears to be greater than predicted from traditional
bioassays with rats.

CHAPTER 3
THE EFFECT OF EXCESSIVE DIETARY VITAMIN A ON PERFORMANCE
AND VITAMIN E STATUS IN SWINE FED DIETS
VARYING IN DIETARY VITAMIN E
Introduction
Both vitamins A and E are fat soluble vitamins. There
is evidence that high dietary vitamin A may interfere with
both vitamin E absorption and blood a-tocopherol
concentration. High dietary vitamin A reduced absorption of
a-tocopherol in trials with chicks (Sklan and Donoghue,
1982). Abawi and Sullivan (1989) noted decreased plasma
vitamin E concentration when chicks received high (100,000
IU/kg) levels of dietary vitamin A. Blakely et al. (1991)
also reported that high dietary vitamin A (100 times
requirement) plus high levels of beta carotene (480 mg/kg of
diet) reduced plasma vitamin E concentration by 77% in
rats.
Limited research is available on the effect of dietary
vitamin A on vitamin E status in pigs. Our study was done
to evaluate the effect of excessive dietary vitamin A on
performance and on serum and tissue concentrations of a-
tocopherol of growing-finishing pigs fed diets supplemented
with varying levels of vitamin E.
27

28
Experimental Procedures
Eighty-four crossbred pigs with an average initial
weight of 26 kg were divided by sex, weight and litter
origin into pens of two pigs each (1 barrow, 1 gilt). Each
pen was assigned to one of six dietary treatments within
each of seven replications. The treatments for the 2x3
trial consisted of a basal corn-soybean meal diet
supplemented with DL-a-tocophery1 acetate (Hoffmann-La Roche
Inc., Nutley, NJ) at levels of 0, 15, or 150 IU/kg, and
retinyl acetate (Hoffmann-La Roche Inc., Nutley, NJ) at
levels of 2,000 or 20,000 IU/kg of diet. Pigs were fed a
grower diet (Table 3-1), formulated to meet NRC (1988)
requirements (except for vitamins A and E), until they
reached an average body weight of 57 kg and continued on a
finisher diet (Table 3-1) with the same treatment until they
reached an averaged body weight of 107 kg. The vitamin
premix used did not contain vitamins A and E. Pigs were
given feed and water ad libitum. Pigs were housed in an
open-sided building in pens with solid concrete floors.
Pigs were weighed at the start of the feeding phase and
biweekly thereafter. Average daily weight gains, feed to
gain ratios, and average daily feed intakes were determined.
Prior to the start of the study the pigs were fed a nursery
diet that contained 2750 IU of added vitamin A and 22 IU
added vitamin E/kg of feed. The trial was carried out

29
Table 3-1. Composition of diets (%)
Ingredient
Grower
Finisher
Ground corn
75.00
82.05
Soybean meal (48%)
22.00
15.00
Dynafos
1.70
1.70
Ground limestone
. 80
.80
Salt
.25
.25
Trace mineral premixa
. 10
. 10
Vitamin premix
. 10b
. 05c
Se premixd
. 05
. 05
“Provided 200 ppm zinc, 100 ppm iron, 55 ppm manganese,
11 ppm copper, and 1.5 ppm iodine.
bProvided 4.4 mg riboflavin, 22 mg niacin, 18 mg
pantothenic acid, 300 mg choline chloride, 22 ug
vitamin B12, 3 mg vitamin K, and 880 IU vitamin D3 per
kg of diet.
“Provided 2.2 mg riboflavin, 11 mg niacin, 9 mg
pantothenic acid, 150 mg choline chloride, 11 ug
vitamin B12, 1.5 mg vitamin K, and 440 IU vitamin D3 per
kg of diet.
dProvided .1 ppm selenium.

30
during late spring and early summer (March through June).
Pigs were managed following acceptable care and management
practices. Protocol for animal care had been approved by
the University Animal Use Committee.
Blood samples were collected by jugular venipuncture
from each pig at the start (d 0) and on d 3, 7, 21, 35, 63,
and 77 of the feeding period thereafter. Blood samples were
shielded from direct sunlight. Blood samples were
centrifuged after collection, serum harvested, frozen and
stored at -20°C until analyzed. During storage, blood
samples were covered with foil to avoid exposure to light.
Upon termination of the feeding phase, one pig (barrow)
per pen was slaughtered, following accepted slaughter
procedures, at the University of Florida meats laboratory
and tissue samples collected. Tissue samples included
liver, leg (semimembranosus) and neck (rhomboideus) muscle,
back fat (10th rib area) and leaf fat. Tissue samples were
frozen following collection and stored at -20°C until
analyzed.
Vitamin E (a-tocopherol) was extracted from serum
samples using the procedure described by McMurray and
Blanchflower (1979a) with modifications described by Njeru
et al. (1992). Extraction of vitamin E from tissue and feed
samples was done using the procedure of Chung et al. (1992).
This procedure was similar to that of McMurray and
Blanchflower (1979b) and Hatam and Kayden (1979) with

31
modifications (Njeru et al., 1992). Alpha-tocopherol was
determined by injecting 20 ul of the reconstituted sample
(serum, or tissue) into an HPLC (Anderson et al., In Press).
Alpha-tocopherol concentration of samples was calculated
from the known concentration of standards. Spiked samples
of a-tocopherol were found to have a mean recovery rate of
97 ± 3%.
Vitamin A was extracted from serum and tissues as
described by Chew et al. (1991) and Mooney (1992).
Extraction procedures were performed under dark conditions
with either yellow filtered or subdued light. Vitamin A was
assayed and determined by the method of Mooney (1992). The
only modifications were that HPLC prepared samples were
eluted using an 80:20 (vol/vol) mixture of iso-octane:
tetrahydrofuran with 1% acetic acid and retention time was
approximately 10.5 min. All trans retinol (Sigma Chemical
Co., St. Louis, MO) standards were prepared and used to
determine concentration of samples. Several liver samples
were spiked to determine the recovery rate and validate the
extraction procedure. Recovery rate of retinol was found to
be 100 ± 4.7%.
Data collection included serum and tissue
concentrations of vitamin A and E and performance data (body
weight gain, feed-to-gain and feed intake). Tissue data
were log transformed prior to analysis to improve
homogeneity of variance. A univariate repeated measures

32
ANOVA was performed on serum data. Data were analyzed using
the GLM procedure of SAS (1988). Orthogonal polynomial
contrasts were performed to compare treatment means.
Results and Discussion
Growth performance data of the pigs in this study are
summarized in Table 3-2. Pigs grew well on all dietary
treatments (NRC, 1988). An increase in average daily gain
of pigs approached significance (P = .15) in linear fashion
as dietary vitamin E increased; feed to gain was not
affected (P > .1). Dietary vitamin A levels had no effect
on pig performance (P > .1). Hoppe et al. (1992) fed pigs
54 IU of supplemental vitamin E in combination with 5,000,
10,000, 20,000 or 40,000 IU of retinol/kg of diet and also
observed no differences in pig performance. Other
researchers have noted similar results in the chick (Sklan
and Donoghue, 1982; Abawi and Sullivan, 1989) and rat
(Blakely et al., 1991).
Serum a-tocopherol concentrations are reported in
Tables 3-3 and 3-4. Serum tocopherol concentrations were
affected by dietary a-tocopherol on all sampling days except
d 0 (Table 3-4). When no supplemental vitamin E was added
to the diet, there was a steady decline in serum tocopherol
concentration from d 0 to d 77 (Table 3-4). When the diet
was supplemented with 15 IU of a-tocopheryl acetate per kg
of diet, serum tocopherol concentrations declined (P < .05)

33
Table 3-2. Performance of growing-finishing swine fed diets
with different dietary levels of vitamin E and
Vitamin A
Added
Vit. E,
IU/kg
Added vitamin A, IU/kq
2,000 20,000
Mean
SE
A T T 1,, -I »-*
nvy . uaiiy yain, js.y
0
.91
.91
.91
15
.92
. 96
. 94
150
.96
. 92
. 95
Mean
. 93
. 94
. 01
SE
. 02
nvy . uaiiy
0
3.02
3.23
3.13
15
3.10
3.11
3.11
150
3.17
3.08
3.12
Mean
3.09
3.14
. 05
SE
. 09
YYYTrTYr-rTYc —
nvy . iccu/yaiii
0
3.33
3.56
3.44
15
3.39
3.25
3.32
150
3.29
3.32
3.30
Mean
3.34
3.38
. 07
SE
. 08
Note: Seven pens per
treatment with 2 pigs
per pen.
aP>F: E =
.19, A
= .75
, E*A = .22, E linear
= . 15
bP>F: E =
.97, A
= .52
, E*A = .26, E linear
= .94
CP>F: E =
.41, A
= .67
, E*A = .26, E linear
= . 36

34
Table 3-3. Mean a-tocopherol concentrations in blood serum
due to dietary additions of vitamins E and A
Vit. E and A. IU/kqa
Days 0/2b 0/20b 15/2 15/20 150/2 150/20 SE
/Ltg/ml
0
1.14
1.17
1.18
1.24
1.03
1.25
. 11
3
. 60
. 66
.75
.75
2.75
2.04
. 09
7
. 55
. 59
. 92
1.02
3.18
2.92
. 11
21
. 37
. 53
.75
. 84
3.15
2.99
. 10
35
. 39
.45
. 87
.78
3.18
2.99
. 09
63
.31
.32
. 99
.78
3.29
3.24
. 08
77
.33
.31
.98
.70
2.84
3.02
. 11
Note; Each mean is based on 14 observations.
“Vitamin E linear effect P < .01 all days except d 0.
E x A P < .01 d 3 and 21 only.
b2 = 2,000; 20 = 20,000 IU of retinyl acetate.

35
Table 3-4. Main means of serum a-tocopherol due to dietary
additions
of
vitamins
E and A
Days
Vit.
E“,
IU/ka
Vit. Ab.
IU/ka
SE
0
15
150
SE
2,000
20,000
/-¿cj j m X
0
1.15 1
.21
1.14
. 07
1.11
1.22
. 06
3
. 63
.75
2.40
. 06
1.37
1.15
. 05
7
. 57
.97
3.05
. 08
1.55
1.51
. 06
21
.45
. 79
2.92
. 07
1.42
1.35
. 06
35
.41
. 83
3.08
. 06
1.48
1.40
. 05
63
.31
. 88
3.26
. 05
1.53
1.45
. 04
77
.31
.84
2.93
. 08
1.38
1.34
. 06
Note: Each mean is based on 28 or 42 observations.
“Vitamin E linear effect P < .01 all days except d 0; E
quadratic P < .02, d 63 and 77 only.
bVitamin A effect P < .01, d 3 only.

36
from their initial concentration by d 3 and then stabilized
throughout the remainder of the trial (Table 3-4). Serum
tocopherol concentration increased (P < .01) with the
highest vitamin E supplementation level by d 3 and continued
to increase (P < .01) to d 7 after which serum concentration
was maintained throughout the study (Table 3-4). Serum
tocopherol was highest (P < .01) at the highest level of
vitamin E supplementation on all days except d 0. Overall,
as dietary levels of supplemental vitamin E increased, serum
concentration of tocopherol also increased (linear; P <
.01). Other studies have shown dietary vitamin E compounds
are effective in increasing blood tocopherol and that blood
concentrations increased with increasing dietary vitamin E
(Jensen et al., 1988; Asghar et al., 1991 and Anderson et
al., In Press).
Serum tocopherol concentrations due to supplementing
the diet with vitamin A at 2,000 or 20,000 IU/kg of diet are
summarized in Table 3-4. Pigs fed the low level of vitamin
A tended to have higher concentrations of serum tocopherol
than pigs fed the high level of vitamin A on all blood
sampling days, but was significant only on d 3 (P < .01).
Among the three dietary levels of vitamin E, the highest
dietary vitamin E tended to be decreased by the high dietary
vitamin A to the greatest extent, but was significantly
lower (P < .01) on d 3 and d 21 only (Table 3-3). Although
significant differences were noted above, the magnitude of

37
these changes was very small. Therefore vitamin A level of
20,000 IU/kg had only a minimal effect on blood tocopherol
in swine. Weaver et al. (1989), in a trial with young
starting pigs, observed that high dietary levels of vitamin
A (9,900 and 16,500 IU/kg) fed with 33 IU of added vitamin
E/kg tended to lower plasma tocopherol concentrations
slightly. In trials with chicks and rats, however, a
decrease in blood tocopherol was noted upon feeding diets
with excessive vitamin A. In these trials, very high
vitamin A concentrations (> 100,000 IU/kg of diet) were fed
(Sklan and Donoghue, 1982; Blakely et al., 1991). This
negative effect is thought to be attributed to competition
for absorption sites in the small intestine or enhanced
oxidation of tocopherol prior to tocopherol reaching the
small intestine (Sklan and Donoghue, 1982).
Although some differences were observed on individual
sampling days, supplemental vitamin A had no effect (P >
.10) on serum retinol (Table 3-5). Differences that were
noted were very small and within normal values for serum
retinol usually found in the pig (Kaneko, 1980). Blood
retinol was also not affected by dietary supplementation of
various levels of vitamin A in studies done by other
researchers (Abawi and Sullivan, 1989; Blakely et al., 1991
Hoppe et al., 1992).
With two exceptions, there was no effect (P > .10) on
serum retinol due to supplementation of vitamin E at any of

38
Table 3-5. Main means of serum retinol due to dietary
additions of vitamins E and A
Vit. Ea. IU/kcr
Days 0 15 150
Vit, Ab, iu/ka
SE
2,000
20,000 SE
Mg/ml
0
. 34
.34
. 39
. 02
.31
.41
CM
o
•
3
.31
.33
. 35
. 01
.30
.35
. 01
7
. 34
. 32
.35
. 01
.33
.34
. 01
21
. 34
. 36
.35
.01
.35
.35
.01
35
. 37
. 34
. 32
. 01
.35
. 34
. 01
63
.39
.35
. 37
. 01
. 38
.36
. 01
77
. 47
.49
.46
. 02
. 52
.44
. 02
Note: Each mean is based on 28 or 42 observations.
“Vitamin E linear effect P < .05 on d 3 and 35 only.
bVitamin A effect P < .01 on d 3 and 77 only.

39
the dietary levels evaluated in this study. Increasing
dietary vitamin E (Table 3-5) resulted in higher (P < .02)
retinol serum concentration on d 3 in pigs fed the highest
supplemental vitamin E, but also resulted in the lowest (P <
.02) serum retinol concentration on d 35. Although both
were significant, the differences were small. Weaver et al.
(1989) in trials with growing pigs also noted that plasma
vitamin A was not affected by dietary level of vitamin E.
Tissue a-tocopherol increased (linear; P < .001) as
dietary vitamin E increased (Table 3-6). As dietary vitamin
E increased from 15 to 150 IU per kg of diet, tissue
tocopherol increased by at least a factor of 2 or more in
all tissues evaluated. This finding is in agreement with
others who have observed similar responses in pigs (Jensen
et al., 1988; Asghar et al., 1991). Among the tissues
sampled, the highest concentration of a-tocopherol due to
treatment was found in the adipose tissue followed by liver
and muscle tissue, respectively (Table 3-6).
High supplementation of vitamin A had no effect (P >
.1) on tocopherol concentration in any of the tissues
studied (Table 3-6). The retinol concentration in the liver
was greatly enhanced (P < .001) when pigs were fed the high
level of vitamin A (Table 3-7). Retinol concentration in
other tissues (back fat, leaf fat, rhomboideus and
semimembranosus) was not detected and are not reported.
Dietary vitamin E had no effect (P > .10) on liver retinol

Table 3-6. Main means of tissue a-tocopherol concentrations due to dietary
additions of vitamins E and A
Tissue
Vit. Ea, IU/kcr
0 15 150
SE
Vit. A. IU/kq
2,000 20,000
SE
-Mg/g-
Semimembranosus
. 4a
1.4b
4.0C
. 14
1.9
2.0
. 11
Rhomboideus
. 3a
2.0b
6.5C
.30
2.9
2.9
.24
Back fat
1. Ia
2.7b
11.0C
.35
5.2
4.5
.28
Leaf fat
. 9a
3 . lb
o
o
o
rH
. 26
4.8
4.5
.21
Liver
1.0a
2.6a
7.2b
. 65
3.8
3.5
. 53
Note: Each mean is based on 14 or 21 observations.
aVitamin E linear effect P < .001 for all tissues.
-p-
o

41
Table 3-7. Main means of tissue retinol concentrations
due to dietary additions of vitamins E and A
Tissue
Vit. E. IU/kcr
0 15 150
Vit. Aa, IU/ka
SE
2,000 20,000 SE
Liver
503 470
Mg/g.
496 44
103
876
36
Note: Each mean is based on 14 or 21 observations.
“Vitamin A effect P < .001.

42
concentration (Table 3-7). Hoppe et al. (1992) found that
liver retinol was linearly related to dietary retinol in a
trial involving young, growing pigs as was also observed in
the present study. They also found that 10,000 IU of
dietary retinol did not affect tissue a-tocopherol other
than the heart in which a-tocopherol was slightly depressed.
However, Pudelkiewicz et al. (1964) feeding vitamin A
depleted chicks 0, 1,453, 14,535, 1,453,488 and 14,534,883
IU of vitamin A acetate/kg of diet observed a marked decline
in tocopherol concentration in liver tissue at the highest
dietary vitamin A levels.
In conclusion, there was no consistent evidence that
excessive dietary vitamin A (20,000 IU/kg of diet) affected
growth performance or serum or tissue a-tocopherol in
growing-finishing pigs fed diets supplemented with varying
levels of vitamin E. Likewise serum retinol was not
affected by dietary vitamin E.
Summary
A 2 X 3 factorial experiment was conducted to evaluate
excessive dietary vitamin A on vitamin E status and
performance of growing-finishing pigs fed diets supplemented
with varying levels of vitamin E. Treatments consisted of
corn-soybean meal based diets supplemented with DL-a-
tocopheryl acetate to provide 0, 15 or 150 IU added vitamin
E/kg and with retinyl acetate to provide 2,000 or 20,000 IU

43
vitamin A/kg of diet. The trial involved 84 crossbred pigs
(26 kg) divided by sex, weight, and genetic background into
pens of two pigs each. Treatment was assigned at random to
pens within each of seven replications. Pigs were fed
grower diets (.75% lysine) until they reached 57 kg average
weight and were then switched to finisher diets (.60%) until
107 kg. Serum was collected on day 0, 3, 7, 21, 35, 63, and
77 of the feeding period. Tissue samples (liver, muscle,
back fat and leaf fat) were collected from one pig (barrow)
in each pen at the end of the feeding phase. Overall
average daily gain and feed-to-gain were .93 kg and 3.35
respectively, without treatment differences (P > .1).
Excessive dietary vitamin A had no effect (P > .1) on serum
retinol concentrations except d 3 in which there was a small
(P < .01) increase. Serum tocopherol was increased (P <
.01; linear) by d 3 with dietary vitamin E supplementation
and was maintained (P < .01) throughout the feeding period.
High dietary vitamin A resulted in a small but significant
(P < .01) decrease in serum tocopherol on d 3; serum
tocopherol concentrations were not affected on other days.
Tissue tocopherol was increased (P < .001; linear) as
dietary vitamin E increased from 15 to 150 IU/kg. Liver
retinol increased (P < .001) by a factor of eight. No
consistent evidence was found that high dietary vitamin A
interfered with performance or with serum or tissue
tocopherol in growing-finishing swine.

CHAPTER 4
EFFECT OF INJECTED VITAMIN A AND DIETARY SUPPLEMENTATION OF
VITAMIN E ON REPRODUCTIVE PERFORMANCE AND TOCOPHEROL STATUS
IN GESTATING GILTS
Introduction
Supplemental vitamin A and/or B-carotene given via
injection just before and/or shortly after breeding appears
to enhance reproductive performance of gilts and sows (Brief
and Chew, 1985; Coffey and Britt, 1993). There is evidence
that high dietary vitamin A may interfere with both vitamin
E absorption and blood a-tocopherol concentrations. High
dietary vitamin A reduced absorption of a-tocopherol in
trials with chicks (Sklan and Donoghue, 1982). Abawi and
Sullivan (1989) noted a decrease in plasma vitamin E
concentrations when depleted chicks were administered high
(100,000 IU/kg) levels of dietary vitamin A. Blakely et al.
(1991) also reported that high levels of vitamin A (100
times reguirement) plus high levels of B-carotene (480 mg/kg
of diet) reduced vitamin E plasma concentration by 77% in
rats.
This study was conducted to evaluate the effect of
injecting vitamin A just before, during and shortly after
breeding, and dietary supplementation of vitamin E on
reproductive performance and on blood and tissue
44

45
concentrations of a-tocopherol during early gestation of
gilts.
Experimental Procedures
The trial was a 2 x 2 factorial design and involved 32
(7 to 8 month old) crossbred gilts. The gilts were divided
into pens of 8 gilts each. Gilts used in this study were
from a previous trial that involved the feeding of diets
supplemented with either 2,000 (L) or 20,000 (H) IU vitamin
A/kg of diet. This was taken into consideration in the
allotment of gilts to treatment (4L and 4H per pen). Each
pen was randomly assigned to one of four treatments.
Treatments consisted of a basal corn soybean meal diet
(Table 4-1) supplemented with DL-a-tocophery1 acetate
(Hoffmann-La Roche Inc. Nutley, NJ) at levels of either 25
or 500 IU/kg of diet. Gilts were fed experimental diets
beginning 7 d prior to breeding through d 25 of gestation.
Half of the gilts were given three injections (i.m. in the
neck) of 350,000 IU each of vitamin A (vitamin A palmitate
Hoffmann-La Roche Inc. Nutley, NJ); the other half were
injected with vehicle only. The gilts were injected at 7 d
prebreeding (d -7), at breeding (d 0) and 7 d postbreeding
(d 7). Gilts were fed 1.9 kg of feed/hd once daily and
given free access to water. Gilts were housed in an open¬
sided building with solid concrete floors. Gilts were
checked twice daily for estrus during the trial and doubled

46
Table 4-1. Composition of diet
fed to gestating gilts
Ingredient
%
Ground corn
84
. 66
Soybean meal (48%)
12
.40
Dynafos
1
. 54
Ground limestone
. 65
Salt
. 50
Trace mineral”
. 10
Vitamin premixb
. 10
Se premixc
. 05
“Provided 200 ppm zinc,
11 ppm copper, 1.5 ppm
100 ppm
iodine,
iron,
and 1
55 ppm manganese,
ppm cobalt.
bProvided 4.4 mg riboflavin, 22 mg niacin, 18 mg
pantothenic acid, 300 mg choline chloride, 22 ug vitamin
B,2, 3 mg vitamin K, 880 IU vitamin D3, and 4000 IU
vitamin A per kg of diet.
cProvided .1 ppm selenium.

mated on their second or third observed estrus to duroc x
hampshire x yorkshire boars.
Blood samples were collected by jugular vein puncture
from each gilt on d -7, 0, 7 and 24 of gestation to monitor
the serum a-tocopherol (vitamin E) and retinol (vitamin A)
concentrations. Blood samples were covered with foil to
prevent exposure to direct sunlight, taken to the
laboratory, centrifuged and the serum harvested. Serum was
stored at -20°C until analyzed for a-tocopherol and retinol
concentration. During storage, serum samples were covered
with foil to prevent exposure to light.
Gilts were slaughtered, following accepted slaughter
procedures, on d 25 of gestation at the University of
Florida meats laboratory. Reproductive tracts were
immediately removed and refrigerated for later counting of
corpora lútea (CL) and recovery of embryos. Tissue samples
were also collected which consisted of endometrium, embryo,
ovary, uterus, liver, leaf fat, back fat and muscle
(semimembranosus and rhomboideus). Tissue samples were
stored at -20°C until analyzed for a-tocopherol and retinol
concentrations.
The trial was carried out during the summer and early
fall (July through October). Pigs were managed following
acceptable care and management practices throughout the
study. Protocol for animal care had been approved by the
University Animal Use Committee. Three pigs from different

48
treatment groups were eliminated from the study due to
death, lameness and failure to conceive.
Alpha-tocopherol was extracted from serum and tissues
using procedures as described earlier (Anderson et al., In
Press). Alpha-tocopherol concentration was determined by
injecting 10 ul of the extracted sample (serum and tissue)
into the HPLC system. Retinol was extracted, assayed and
concentration determined from the serum and tissues by the
method as described previously (Mooney, 1992).
Experimental data included serum and tissue
concentrations of a-tocopherol and retinol, and reproductive
performance. Tissue data was log transformed prior to
analysis to improve homogeneity of variance. A univariate
repeated measures ANOVA was performed on serum data (SAS,
1988). Data were analyzed as a 2 X 2 factorial design with
the factors being dietary vitamin E level and whether or not
gilts were injected with vitamin A.
Results and Discussion
Levels of vitamin E used were chosen to reflect NRC
(1988) requirement and to give a very high level. The
injected level for vitamin A was chosen because it was
thought to be the upper limit that would elicit a response
and not be toxic to the gilts (also some evidence of this
dosage was being used).
Reproductive performance data are summarized in Table

49
Table 4-2. Mean reproductive response criteria of gestating
gilts given dietary additions of vitamin E and injected
with vitamin A
Item
Vit. E
, IU/kg
and Vit. A
ini .“
SE
25/No
25/Yes
500/No
500/Yes
No. of CL
14
13
15
15
.76
No. of Embryob
12
13
12
15
1.13
Embryo wt., g
9
9
7
8
. 98
Ovary wt., g
13
13
14
15
.90
Uterus wt., kg
3
2
2
2
. 22
Note: Each mean
is based
on 7 or
8 observations.
“Three injections of 350,000 IU each.
bEffect of vitamin E (P = .14), effect of
vitamin A (P = .13).

50
4-2. Although none of the differences noted with
reproductive data were significant (P > .1), due to the
small number of gilts per treatment and the inherent nature
of swine reproductive data, nevertheless, some positive
trends were observed due to treatment. Gilts receiving the
high vitamin E and high vitamin A treatment had larger (P =
.16) litters than gilts given other treatments. Embryonic
survival was 86% in gilts given the low dietary vitamin E
with no injected vitamin A and 80% in gilts given the high
dietary vitamin E with no injected A. Embryonic survival
was 100% in gilts injected with vitamin A and fed either the
low or high vitamin E diets. Brief and Chew (1985) reported
larger litter size and higher embryonic survival in gilts
receiving weekly injections of vitamin A (12,300 IU) and B-
carotene (33 mg) compared to gilts fed vitamin A and B-
carotene at the same levels. However, these gilts in the
study of Brief and Chew (1985) were depleted of vitamin A
and B-carotene for 5 weeks before the start of the study.
Coffey and Britt (1993) observed on average .5 pig increase
in the number of pigs born alive and higher embryonic
survival in sows given i.m. injections of vitamin A
palmitate (50,000 IU) compared to sows given vehicle only
(corn oil) on day of weaning, mating and 7 d postbreeding.
These sows were also supplemented with 11,000 IU of vitamin
A acetate per kg of diet.
Serum concentrations of a-tocopherol in gilts fed diets

51
supplemented with two levels of vitamin E with and without
injected vitamin A are reported in Tables 4-3 and 4-4.
Initial a-tocopherol serum concentrations in gilts were
similar (d -7). When dietary levels were increased from 25
to 500 IU of DL-a-tocopheryl acetate per kg of diet, serum
a-tocopherol concentrations increased (P < .01) by d 0 and
were maintained throughout the duration of the study (Table
4-4). This finding is in agreement with other studies that
have shown that dietary vitamin E compounds are effective in
increasing serum tocopherol, and that serum concentration
increased with increasing dietary vitamin E (Jensen et al.,
1988; Asghar et al., 1991; Mahan, 1991 and Anderson et al.,
In Press).
Serum a-tocopherol was not affected (P > .1) by
injecting gilts with vitamin A (retinyl palmitate) except on
d 7 (Table 4-4). On d 7, the gilts fed the low vitamin E
diet had similar (P > .1) serum tocopherol concentrations
regardless of vitamin A treatment, whereas gilts fed the
high vitamin E diet had higher (P < .08) serum tocopherol
concentrations when injected with vitamin A than gilts not
injected with vitamin A (Table 4-3).
Serum concentrations of retinol due to treatment are
shown in Tables 4-5 and 4-6. In general, there was no
consistent effect on serum retinol due to dietary vitamin E
supplementation level or injection of vitamin A. However, a
difference in serum retinol (P < .08) was observed on d 0 in

52
Table 4-3. Mean serum a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin
E and injected with vitamin A
Sampling
day
Vit.
E. IU/kcf
and vit.
A ini . “bc
25/No
25/Yes
500/No
500/Yes
SE
_ z _ ^
/i <3 / ^ 1
-7
. 8
. 9
i. i
. 6
. 17
0
1.2
1.2
3.6
3.6
. 18
7
1.1
1.1
3.4
4.0
. 15
24
1.3
1.3
3.8
3.6
. 15
Note; Each mean is based on 7 or 8 observations.
“Three injections of 350,000 IU each.
bEffect of vitamin E (P < .01), d 0, 7, 24.
CE x A effect (P < .08), d 7.

53
Table 4-4. Main mean serum a-tocopherol concentrations
in gestating gilts given dietary additions of
vitamin E and injected with vitamin A
Sampling
day
Vit. E
. IU/ka“
Vit.
A ini .bc
SE
25
500
No
Yes
fjuy-j / mi
-7
.8
. 9
. 9
. 8
. 12
0
1.2
3.6
2.4
2.4
. 13
7
1.1
3.7
2.3
2.6
. 11
24
1.3
3.7
2.5
2.5
. 11
Note: Each
mean is
based on 14
, 15 or 16 observations.
“Effect of
vitamin
E (P < .01)
d 0,
7 and 24.
bThree injections of 350,000 IU each.
cEffect of vitamin A (P < .06), d 7 only.

54
Table 4-5. Mean serum retinol concentrations in gestating
gilts given dietary additions of vitamin E and
injected with vitamin A
Sampling
day
Vit
E. IU/kaa
and vit.
A ini .bc
SE
25/No
25/Yes
500/No
500/Yes
jtxcj / ml
-7
. 51
. 55
. 52
. 51
. 03
0
.44
.49
. 51
. 59
. 03
7
.40
.43
. 47
.42
. 03
24
.46
.47
.48
.45
. 03
Note: Each
mean is
based on 7
or 8 observations.
“Effect of
vitamin
E
(P < .03)
, d 0 only.
bThree injections of
350,000 IU each.
cEffect of
vitamin
A
(P < .08)
, d 0 only.

55
Table 4-6. Main mean serum retinol concentrations in
gestating gilts given dietary additions of vitamin
E and injected with vitamin A
Sampling
day
Vit.
E. IU/ka“
Vit. A
ini .b-c
SE
25
500
NO
Yes
.
fj/ cf ¡ m -L
-7
.53
. 52
. 52
. 53
. 02
0
.46
. 55
.47
. 54
. 02
7
.41
.44
.43
.42
. 02
24
.46
.47
.47
.46
. 01
Note: Each mean is based on 14, 15 or 16 observations.
“Vitamin E effect (P < .03) d 0 only.
bThree injections of 350,000 IU each.
cVitamin A effect (P < .08) d 0 only.

56
that serum retinol concentration was highest in gilts fed
the high vitamin E and injected with vitamin A, and lowest
in gilts fed low vitamin E without injected A. The
difference noted was very small and within normal values for
serum retinol concentrations usually found in the pig. Our
findings agree with Weaver et al. (1989) in that plasma
vitamin A concentration was not affected by dietary level of
vitamin E. Serum retinol concentrations in the chick and
rat have also been reported not to be affected by dietary
supplementation of vitamin E in studies done by Abawi and
Sullivan, (1989) and Blakely et al. (1991), respectively.
Mooney (1992) injected gilts with vitamin A palmitate
(ranging from 53,200 to 106,400 IU given once weekly), or 6-
carotene (106.4 to 425.6 mg) and observed no difference in
plasma concentrations of either retinol or 6-carotene. In
contrast, Brief and Chew (1985) noted increased plasma
vitamin A concentration upon injecting vitamin A, however,
gilts used in their research were depleted of vitamin A
prior to the study and they also received injected 6-
carotene. Serum retinol may have been elevated with
injection of vitamin A early in our study but may have been
missed since blood samples were taken 7 d after injection.
Tissue a-tocopherol concentration in gestating gilts
increased (P < .01) as dietary supplementation of vitamin E
increased (Table 4-7) in all tissues except adipose. This
finding is in agreement with others who have observed

57
Table 4-7. Mean tissue a-tocopherol concentrations of
gestating gilts given dietary additions of vitamin E
and injected with vitamin A
Tissue3
Vit.
E, IU/kg
and vit.
A inj.b
25/No
25/Yes
500/No
500/Yes
SE
mu/g
Liver
4
4
24
23
1.6
Back fat
6
8
9
9
1.5
Leaf fat
9
10
12
12
2.2
Semimembranosus
2
3
4
4
. 3
Rhomboideus
3
3
7
7
. 6
Endometrium*1
2
1
4
5
. 3
Embryo
. 4
. 4
. 8
. 7
. 05
Oviduct
1
1
4
3
. 3
Uterus
1
1
4
4
. 2
Ovary
20
19
104
90
5.6
Note: Each mean
is based
on 7 or
8 observations.
“Effect of vitamin E (P < .01) for all tissues except
back fat and leaf fat.
bThree injections of 350,000 IU each.
cWet tissue basis.
dEffect of vitamin A (P < .08); E x A (P < .04).

58
similar responses in the pig (Jensen et al., 1988; Asghar et
al., 1991; Mahan, 1991). Average tocopherol concentration
increased by a factor of 2 in embryos upon high dietary
supplementation indicating that tocopherol is transferred
from the dam to the developing embryo. Vitamin A injections
had no effect (P > .1) on tissue a-tocopherol concentrations
except in the endometrium where there was a vitamin E x
vitamin A interaction (P < .04). In the endometrium gilts
fed low vitamin E and injected with vitamin A had slightly
lowered tocopherol concentration, while gilts fed the high
vitamin E and injected with vitamin A had increased
tocopherol concentration over that of the non injected gilts
(Table 4-7). Vitamin A injections appear to have no effect
on the transfer of tocopherol into the developing embryos as
tocopherol concentration in the embryos was not influenced
by vitamin A injection (P > .1).
Injecting vitamin A had no effect (P > .1) on retinol
concentration in any of the tissues studied including the
liver. Retinol concentrations in tissues other than liver,
however, were very small or nonexistent. Average
concentration in the liver was 386 ^g/g. Mooney (1992)
found no differences in concentration of retinol in uterine
flushings in gilts that were injected with varying levels of
vitamin A; liver retinol was not determined.
Among the tissues sampled, the highest average
concentration of a-tocopherol upon supplementation of high

59
level of vitamin E was found in the ovary, followed by
liver, adipose, rhomboideus and endometrium, respectively
(Table 4-8). Three other tissues followed (semimembranosus,
oviduct, uterus) that had similar average concentrations and
embryo tissue had the lowest a-tocopherol concentration
(units per wet tissue basis).
In conclusion, there was no consistent evidence found
in this study that injecting a relatively large amount of
vitamin A (3 injections of 350,000 IU) just before, during
and shortly after breeding, significantly improved
reproductive performance, or interfered with serum or tissue
concentrations of a-tocopherol in gestating gilts fed diets
supplemented with 25 or 500 IU of vitamin E/kg of diet.
However, tocopherol concentration was increased further in
the endometrial tissue when vitamin A was given along with
high dietary vitamin E. No evidence was found that
injections of vitamin A interfered with the transfer of a-
tocopherol to the developing embryo. Likewise, serum
retinol concentrations were not affected by treatment.
Summary
A 2 x 2 factorial experiment was conducted to determine
the effects of injecting vitamin A and feeding vitamin E on
reproductive performance and on blood serum and tissue
concentrations of tocopherol during early gestation of
gilts. Thirty-two crossbred gilts were fed corn soybean-

60
Table 4-8. Main mean tissue a-tocopherol concentrations in
gestating gilts given dietary additions of vitamin E
and injected with vitamin A
Tissues3
Vit.
E, IU/kg
Vit.
A inj.b
25
500
No
Yes
SE
M<3/g â– 
Liver
4
24
14
14
1.1
Back fat
7
9
7
9
1.0
Leaf fat
9
12
10
11
1.5
Semimembranosus
3
4
3
3
. 2
Rhomboideus
3
7
5
5
. 4
EndometriumJ
1.4
4.6
2.8
3.3
. 18
Embryo
.4
. 8
. 6
. 6
. 03
Oviduct
1
4
3
2
.2
Uterus
1
4
3
3
. 1
Ovary
19
97
61
55
3.8
Note: Each mean is
based
on 14, 15
or 16
observations.
“Effect of vitamin E (P < .01) for all tissues except
back fat and leaf fat.
bThree injections of 350,000 IU each.
cWet tissue basis.
dEffect of vitamin A (P < .08).

61
meal based diets supplemented with DL-a-tocophery1 acetate
to provide either 25 or 500 IU of added vitamin E/kg of
diet. Gilts were fed daily 1.9 kg/h beginning d -7
prebreeding through d 25 of gestation. Half of the gilts
were injected (i.m.) with 350,000 IU of retinol palmitate at
d -7 prebreeding, breeding (d 0) and d 7 postbreeding; the
other half were injected with vehicle only. All gilts were
double mated during their second or third estrus. Blood
samples were collected on d —7, 0, 7 and 24 of gestation.
Gilts were slaughtered on d 25 of gestation following
accepted slaughter procedures. Twenty-nine gilts conceived.
The number of corpora lútea and embryos was not affected (P
> .1) by treatment. Serum tocopherol concentrations
increased with 500 IU of vitamin E by d 0 and were stable
through d 24 of gestation (P < .01). Vitamin A injections
had no effect (P > .1) on serum tocopherol concentrations
except on d 7 when a small increase (P < .06) was noted.
High dietary vitamin E increased tocopherol concentration (P
< .01) in all tissues examined except fat. A vitamin E x
vitamin A interaction (P < .04) was noted in endometrium
tissue. Low dietary vitamin E and injections of vitamin A
slightly lowered tocopherol concentration, while high
vitamin E and vitamin A injections increased tocopherol
concentration in the endometrium. Increasing dietary
vitamin E increased serum and tissue tocopherol
concentrations. Vitamin A injections had little or no

62
effect on these concentrations during early gestation of
gilts.

CHAPTER 5
GENERAL CONCLUSIONS
Three experiments were conducted, one to determine the
bioavailability of four forms of vitamin E compounds, and
two to assess the effect of high levels of vitamin A on the
vitamin E status of growing finishing pigs or gestating
gilts.
In experiment 1 the biopotency of four forms of vitamin
E were determined. Generally, the acetate forms resulted in
greater serum and tissue concentrations of vitamin E (a-
tocopherol) than the alcohol forms, due to the greater
stability of the acetate forms that was noted in mixed feed.
Serum tocopherol increased rather rapidly when the four
compounds were fed. Dietary supplementation of D-a-
tocopheryl acetate resulted in the highest serum tocopherol
throughout the study, compared to concentrations obtained
for pigs fed the other compounds indicating a greater
biopotency (IU/mg) for swine than determined by the
traditional rat bioassay. A similar trend was observed with
tissue (liver, back fat, leaf fat, and muscle) tocopherol
concentrations as with serum concentrations, with the liver
having the highest concentration. In general, all forms
would probably be suitable dietary supplemental sources if
63

64
the stability of the alcohols were improved. Encapsulating
the alcohol forms to protect them from destruction would
increase their suitability for use in mixed feed.
Experiment 2 was conducted to evaluate the effect of
feeding excessive vitamin A on growth performance, and on
blood and tissue a-tocopherol (vitamin E) levels of growing¬
finishing pigs. High dietary vitamin A (20,000 IU/kg of
diet) was found not to affect or have little affect on pig
performance, or on blood or tissue concentrations of a-
tocopherol. A threefold increase (P < .01) in serum
tocopherol occurred on all sampling days when dietary
supplementation increased from 15 to 150 IU/kg. Tissue
tocopherol also increased (P < .001) as dietary vitamin E
increased from 15 to 150 IU/kg. Tissue tocopherol
concentration increased (P < .001) by a factor of at least
two in all tissues evaluated. Liver retinol increased (P <
.001) eightfold with a tenfold increase in dietary vitamin
A. Even in the liver when vitamin A (retinol) concentration
was high, a-tocopherol concentration was not affected.
Thus, the form of vitamin A within the liver is not in a
form which can lead to oxidative destruction of a-tocopherol
or the concentrations encountered may not have been high
enough to affect a-tocopherol.
Experiment 3 evaluated the effect of giving a high
level of vitamin A via intramuscular injections on
reproductive performance, and on serum and tissue a-

65
tocopherol concentrations during early gestation of gilts.
High levels of vitamin A (350,000 IU per week) did not
affect reproductive performance, or serum or tissue
concentrations of a-tocopherol or retinol in this study. As
observed in the previous studies, increasing dietary levels
of vitamin E increased blood serum and tissue tocopherol
concentrations including reproductive tissues. Alpha-
tocopherol concentration also increased in the embryo when
dietary vitamin E was increased. The increased
concentration of a-tocopherol indicates a transfer of
tocopherol from the dam to the developing embryo.
Fluctuations of retinol concentrations in serum may have
been missed due to weekly sampling of blood. Blood sampling
on d 2, 3, or on a more frequent routine after vitamin A
injections would provide more answers.
Further research would be desirable to determine
vitamin E bioavailability other than by oral administration.
Fecal sample tocopherol analysis may provide more
information from which better conclusions might be drawn
concerning digestion and absorption. Studies using larger
numbers of gilts and/or sows is recommended to determine if
vitamin A injection alone or in combination with high
vitamin E, either orally or by injection, would enhance
reproductive performance.

REFERENCES
Abawi, F. G., and T. W. Sullivan. 1989. Interaction of
vitamins A, D3/ E, and K in the diet of broiler chicks.
Poult. Sci. 68:1490.
Abawi, F. G., T. W. Sullivan, and S. E. Scheideler. 1985.
Interaction of dietary fat with levels of vitamin A and
E in broiler chicks. Poult. Sci. 64:1192.
Adams, C. R. 1978. Vitamin product forms for animal
feeds. In: Vitamin Nutrition Update, Seminar Series
2. Roche Publ. Nutley, NJ.
Ames, S. R. 1979. Biopotencies in rats of several forms
of alpha-tocopherol. J. Nutr. 109:2198.
Anderson, L. E.,Sr., R. 0. Myer, J. H. Brendemuhl, and L.
R. McDowell. Bioavailability of various vitamin E
compounds for finishing swine. J. Anim. Sci. In press.
Arnrich, L., and V. A. Arthur. 1980. Interaction of fat-
soluble vitamins in hypervitaminoses. Ann. of N. Y.
Acad. Sci. 355:109.
Asghar, A., J. I. Gray, E. R. Miller, P. K. Ku, A. M.
Booren, and D. J. Buckley. 1991. Influence of
supranutritional vitamin E supplementation in the feed
on swine growth performance and deposition in different
tissues. J. Sci. Food Agrie. 57:19.
Baker, H., G. J. Handelman, S. Short, L. J. Machlin, H.
N. Bhagavan, E. A. Dratz, and 0. Frank. 1986.
Comparison of plasma a and gamma tocopherol levels
following chronic oral administration of either all-
rac-a-tocopheryl acetate or RRR-a-tocophery1 acetate in
normal adult male subjects. Am. J. Clin. Nutr. 43:382.
Behrens, W. A., and R. Madere. 1991. Tissue discrimination
between dietary RRR-a- and all-rac-a-tocopherols in
rats. J. Nutr. 121:454.
Blakely, S. R., G. V. Mitchell, M. Y. Jenkins, E.
Grundel, and P. Whittaker. 1991. Canthaxanthin and
66

67
excess vitamin A alter a-tocopherol, carotenoid and
iron status in adult rats. J. Nutr. 121:1649.
Brandner, G. 1971. "Vitamin E." Bell Publishing Company,
New York.
Bratzler, J. W., J. K. Loosli, V. N. Krukousky, and L. A.
Maynard. 1950. Effect of the dietary level of
tocopherols on their metabolism in swine. J. Nutr.
42:59.
Brief, S., and B. P. Chew. 1985. Effects of vitamin E and B-
carotene on reproductive performance in gilts. J.
Anim. Sci. 60:998.
Burton, G. W., K. H. Cheeseman, T. Doba, K. U. Ingold, and
T. F. Slater. 1983. Biology of Vitamin E. Ciba
Foundation Symposium 101. Pitman, London.
Chew, B. P., T. S. Wong, J. J. Michael, F. E. Standaert, and
L. R. Heirman. 1991. Kinetic characteristics of B-
carotene uptake after an injection of B-carotene in
pigs. J. Anim. Sci. 69:4883.
Chung, Y. K., D. C. Mahan, and A. J. Lepine. 1992. Efficacy
of dietary d-a-tocopherol and dl-a-tocophery1 acetate
for weanling pigs. J. Anim. Sci. 70:2485.
Coelho, M. B. 1991. "Vitamin E in Animal Nutrition and
Management." BASF Corp. Parsippany, NJ.
Coffey, M. T., and J. H. Britt. 1993. Enhancement of sow
reproductive performance by B-carotene or vitamin A.
J. Anim. Sci. 71:1198.
Cunha, T. J. 1977. "Swine Feeding and Nutrition."
Academic Press, New York.
Diplock, A. T. 1985. "Fat-Soluble Vitamins." Technomic
Publishing Co. Inc., Lancaster, Pennsylvania.
Dove, C. R., and R. C. Ewan. 1991. Effect of trace
minerals on the stability of vitamin E in swine grower
diets. J. Anim. Sci. 69:1994.
Draper, H. H. 1980. Nutrient relationships. In: Machlin,
L. J. "Vitamin E: A Comprehensive Treatise." Marcel
Dekker, New York.
Erdman, J. W., Jr., C. L. Poor, and J. M. Dietz. 1988.
Factors affecting the bioavailability of vitamin A,
carotenoids, and vitamin E. Food Technol. 42:214.

68
Evans, H. M. 1962. The pioneer history of vitamin E.
Vitam. and Horm. 20:379.
Harris, P. L., and M. I. Ludwig. 1949a. Vitamin E
potency of a-tocopherol esters. J. Biol. Chem.
180:611.
Harris, P. L., and M. I. Ludwig. 1949b. Relative
vitamin E potency of natural and of synthetic a-
tocopherol. J. Biol. Chem. 170:1111.
Hatam, L. J., and H. J. Kayden. 1979. A high-performance
liquid chromatographic method for the determination of
tocopherol in plasma and cellular elements of the
blood. J. Lipid Res. 20:639.
Hidiroglou, N., N. Cave, A. S. Atwal, E. R. Farnworth, and
L. R. McDowell. 1992. Comparative vitamin E
requirements and metabolism in livestock. Ann. Rech.
Vet. 23:337.
Hidiroglou, N., and L. R. McDowell. 1987. Plasma and
tissue levels of vitamin E in sheep following
intramuscular administration in an oil carrier.
Internat. J. Vit. Nutr. Res. 57:261.
Hidiroglou, N., L. R. McDowell, and R. Pastrana. 1988.
Bioavailability of various vitamin E compounds in
sheep. Internat. J. Vit. Res. 58:189.
Hoppe, P. P., F. J. Schoner, and M. Frigg. 1992. Effects of
dietary retinol on hepatic retinol storage and on
plasma and tissue a-tocopherol in pigs. Internat. J.
Vit. Nutr. Res. 62:121.
Horwitt, M. K., W. H. Elliott, P. Kanjananggulpan, and C. D.
Fitch. 1984. Serum concentrations of a-tocopherol after
ingestion of various vitamin E preparations. Am. J.
Clin. Nutr. 40:240.
Howard, K. A., S. V. Radecki, E. R. Miller, A. J. Thulin,
and D. E. Ullrey. 1990. Relative bioavailability of
vitamin E of natural or synthetic origin in growing
pigs. Res. Rep. 502. Michigan State Univ. Agrie.
Experiment Station, East Lansing, MI.
Jensen, M., J. Hakkarainen, A. Lindholm, and L. Jonsson.
1988. Vitamin E requirement of growing swine. J. Anim.
Sci. 66:3101.
Jensen, M., A. Lindholm, and J. Hakkarainen. 1990. The
vitamin E distribution in serum, liver, adipose and

69
muscle tissues in the pig during depletion and
repletion. Acta Vet. Scand. 31:129.
Kaneko, J. J. 1980. "Clinical Biochemistry of Domestic
Animals." (3rd Ed.). Academic Press, New York.
Koch-Weser, J. 1974. Medical intelligence. The New England
J. of Med. 29(5):233.
Kusin, J. A., V. Reddy and B. Sivakumar. 1974. Vitamin E
supplements and the absorption of a massive dose of
vitamin A. Am. J. Clin. Nutr. 27:774.
Machlin, L. J. 1980. "Vitamin E: A Comprehensive
Treatise." Marcel Dekker, New York.
Mahan, D. C. 1991. Assessment of the influence of dietary
vitamin E on sows and offspring in three parities:
reproductive performance, tissue tocopherol, and
effects on progeny. J. Anim. Sci 69:2904.
Mahan, D. C., and A. L. Moxon. 1980. Effect of dietary
selenium and injectable vitamin E-selenium for weanling
swine. Nutr. Rep. Int. 21:829.
Malm, A., E. F. Walker Jr., M. Homan, D. Kirtland, A. Aydin,
and W. G. Pond. 1976. Glutathione peroxidase and other
enzymes in serum of sow and their progeny fed vitamin
E adequate or deficient diets with added Se. Nutr. Rep.
Int. 14:185.
Mason, K. E. 1980. The First Two Decades of Vitamin E
History. In: Machlin, L. J., "Vitamin E: A
Comprehensive Treatise." Marcel Dekker, Inc., New
York.
McDowell, L. R. 1989. "Vitamins in Animal Nutrition."
Academic Press, Inc., San Diego, CA.
McDowell, L. R., J. A. Froseth, R. C. Piper, I. A. Dyer, and
G. H. Kroening. 1977. Tissue selenium and serum
tocopherol concentrations in selenium-vitamin E
deficient pigs fed pea (Pisam sativum). J. Anim. Sci.
45:1326.
McMurray, C. H., and W. J. Blanchflower. 1979a. Application
of a high-performance liquid chromatographic
fluorescence method for the rapid determination of a-
tocopherol in the plasma of cattle and pigs and its
comparison with direct fluorescence and high-
performance liquid chromatography-ultraviolet detection
methods. J. Chromatogr. 178:525.

70
McMurray, C. H., and W. J. Blanchflower. 1979b.
Determination of a-tocopherol in animal feedstuffs
using high-performance liquid chromatography with
spectrofluorescence detection. J. Chromatgr. 176:488.
Mooney, K. 1992. The effects of supplemental vitamin A
or 6-carotene on reproduction and the localization
of 6-carotene during gestation in gilts. M.S. Thesis.
Univ. of Florida, Gainesville.
Njeru, C.A., L. R. McDowell, N. S. Wilkinson, S. B. Linda,
S. N. Williams, and E. L. Lentz. 1992. Serum a-
tocopherol concentration in sheep after intramuscular
injection of DL-a-tocopherol. J. Anim. Sci. 70:2562.
NRC. 1988. "Nutrients Requirements of Swine." (9th Ed.).
National Academy Press, Washington, DC.
Pudelkiewicz, W. J., L. Webster, and L. D. Matterson. 1964.
Effects of high levels of dietary vitamin A acetate on
tissue tocopherol and some related observations. J.
Nutr. 84:113.
Raacke, I. D. 1983. Herbert McLean Evans: a biographical
sketch. J. Nutr. 113(5):929.
SAS. 1988. " SAS User's Guide: Statistics." SAS Inst. Inc.,
Cary, NC.
Scott, M. L. 1969. Studies of vitamin E and related
factors in nutrition and metabolism. In: DeLuca,
H. F., and J. W. Suttie. "The Fat Soluble Vitamins."
The University of Wisconsin Press, Madison.
Sklan, D., and S. Donoghue. 1982. Vitamin E response to
high dietary vitamin A in the chick. J. Nutr. 112:759.
Tappel, A. L. 1962. Vitamin E as the biological lipid
antioxidant. Vitamins and Hormones. 20:493.
"The American Heritage Dictionary." 1982. (2nd Ed.).
Houghton Mifflin, Boston.
Thompson, J. 1993. Vitamin E injections improve pig
survival. Feedstuffs 65(21):13.
Ullrey, D. E. 1981. Vitamin E for swine. J. Anim. Sci.
53:1039 .
Weaver, E. M., G. W. Libal, C. R. Hamilton, and I. S.
Parker. 1989. Relationship between dietary vitamin A
and E on performance and vitamin E status of the weaned
pig. J. Anim. Sci. 67(Suppl. 2):113. (Abstr.).

BIOGRAPHICAL SKETCH
Lee E. Anderson, Sr., was born December 1944 in
Pineland, South Carolina. He is married to Erma L. Anderson
and the father of five children. He graduated from
Middleton Senior High School in 1962. He graduated with an
Associate of Arts degree in 1964 from Gibbs Junior College.
He received a Bachelor of Science in Agriculture in 1969
from Florida A and M University. He received the Master of
Science in reproductive physiology from the University of
Florida in 1972. Since receiving the M.S. he has been
employed at Alcorn State University, Fort Valley State
University and Florida A and M University, in extension,
research and instructional programs. He is presently a
candidate for the Ph.D.
71

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.
Robert 0. Myer,Chairman
Associate Professor of Animal
Science
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.
JM
Joel H. Brendemuhl'
Associate Professor of Animal
Science
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.
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
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.
Lee R. McDowell
Professor of Animal Science

This dissertation was submitted to the Graduate Faculty
of the College of Agriculture and to the Graduate School and
was accepted as partial fulfillment of the reguirements for
the degree of Doctor of Philosophy.
August, 1993
Dean, Graduate School





PAGE 1

9,7$0,1 ( 67$786 ,1 6:,1( $6 $))(&7(' %< )250 25 /(9(/ 2) ',(7$5< 9,7$0,1 ( $1'25 %< 6833/(0(17$7,21 2) 9,7$0,1 $ %\ /(( ( $1'(5621 65 $ ',66(57$7,21 35(6(17(' 72 7+( *5$'8$7( 6&+22/ 2) 7+( 81,9(56,7< 2) )/25,'$ ,1 3$57,$/ )8/),//0(17 2) 7+( 5(48,5(0(176 )25 7+( '(*5(( 2) '2&725 2) 3+,/2623+< 81,9(56,7< 2) )/25,'$ 81,9(56,7< &) )/25,'$ /,%5$5,(6

PAGE 2

7KLV GLVVHUWDWLRQ LV GHGLFDWHG WR P\ ZLIH (UPD DQG FKLOGUHQ 9DOHULH /HH -U -HQD\D &DOYLQ DQG .HOYLQ DQG WR WKH PHPRU\ RI P\ SDUHQWV )ORUHQFH $QGHUVRQ 0\UWLV 6WHSKHUVRQ DQG /HRQ $QGHUVRQ IRU WKHLU ORYH VXSSRUW DQG HQFRXUDJHPHQW

PAGE 3

$&.12:/('*0(176 7KH DXWKRU VLQFHUHO\ DSSUHFLDWHV WKH HIIRUWV RI DOO PHPEHUV RI KLV VXSHUYLVRU\ FRPPLWWHH 'U 5REHUW 0\HU LV JUHDWO\ DSSUHFLDWHG IRU KLV XQGHUVWDQGLQJ JXLGDQFH DGYLFH DQG FRPSDQLRQVKLS RYHU WKH SDVW VHYHUDO \HDUV 6LQFHUH WKDQNV JR WR 'U -RHO %UHQGHPXKO IRU KLV KHOS DQG JXLGDQFH DQG KLV H[SHUWLVH DQG DVVLVWDQFH GXULQJ GDWD FROOHFWLRQ 7KH DXWKRU LV YHU\ DSSUHFLDWLYH RI 'U /HH 0F'RZHOOn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

PAGE 4

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

PAGE 5

7$%/( 2) &217(176 3DJH $&.12:/('*0(176 LLL /,67 2) 7$%/(6 YL $%675$&7 YLLL &+$37(56 ,1752'8&7,21 327(1&< 2) 9$5,286 9,7$0,1 ( &203281'6 )25 ),1,6+,1* 6:,1( ,QWURGXFWLRQ ([SHULPHQWDO 3URFHGXUHV 5HVXOWV DQG 'LVFXVVLRQ 6XPPDU\ 7+( ())(&7 2) (;&(66,9( ',(7$5< 9,7$0,1 $ 21 3(5)250$1&( $1' 9,7$0,1 ( 67$786 ,1 6:,1( )(' ',(76 9$5<,1* ,1 ',(7$5< 9,7$0,1 ( ,QWURGXFWLRQ ([SHULPHQWDO 3URFHGXUHV 5HVXOWV DQG 'LVFXVVLRQ 6XPPDU\ ())(&7 2) ,1-(&7(' 9,7$0,1 $ $1' ',(7$5< 6833/(0(17$7,21 2) 9,7$0,1 ( 21 5(352'8&7,9( 3(5)250$1&( $1' 72&23+(52/ 67$786 ,1 *(67$7,1* *,/76 ,QWURGXFWLRQ ([SHULPHQWDO 3URFHGXUHV 5HVXOWV DQG 'LVFXVVLRQ 6XPPDU\ *(1(5$/ &21&/86,216 5()(5(1&(6 %,2*5$3+,&$/ 6.(7&+ Y

PAGE 6

/,67 2) 7$%/(6 7DEOH 3DJH &RPSRVLWLRQ RI ILQLVKHU GLHW 3HUIRUPDQFH RI ILQLVKLQJ SLJV IHG GLHWV FRQWDLQLQJ YDULRXV YLWDPLQ ( FRPSRXQGV 9LWDPLQ ( DWRFRSKHUROf FRQFHQWUDWLRQV LQ IHHG $GMXVWHG VHUXP YLWDPLQ ( WRFRSKHUROf FRQFHQWUDWLRQV LQ ILQLVKLQJ VZLQH IHG GLHWV VXSSOHPHQWHG ZLWK GLIIHUHQW YLWDPLQ ( FRPSRXQGV 5HODWLYH ELRSRWHQF\ RI YLWDPLQ ( FRPSRXQGV bf $GMXVWHG WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV LQ ILQLVKLQJ VZLQH IHG GLHWV VXSSOHPHQWHG ZLWK GLIIHUHQW YLWDPLQ ( FRPSRXQGV &RPSRVLWLRQ RI GLHWV bf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

PAGE 7

0HDQ UHSURGXFWLYH UHVSRQVH FULWHULD RI JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 0HDQ VHUXP DWRFRSKHURO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 0DLQ PHDQ VHUXP DWRFRSKHURO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 0HDQ VHUXP UHWLQRO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 0DLQ PHDQ VHUXP UHWLQRO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 0HDQ WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV RI JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 0DLQ PHDQ WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ YLL

PAGE 8

$EVWUDFW RI 'LVVHUWDWLRQ 3UHVHQWHG WR WKH *UDGXDWH 6FKRRO RI WKH 8QLYHUVLW\ RI )ORULGD LQ 3DUWLDO )XOILOOPHQW RI WKH 5HTXLUHPHQWV IRU WKH 'HJUHH RI 'RFWRU RI 3KLORVRSK\ 9,7$0,1 ( 67$786 ,1 6:,1( $6 $))(&7(' %< )250 25 /(9(/ 2) ',(7$5< 9,7$0,1 ( $1'25 %< 6833/(0(17$7,21 2) 9,7$0,1 $ %\ /HH ( $QGHUVRQ 6U $XJXVW &KDLUPDQ 5 0\HU 0DMRU 'HSDUWPHQW $QLPDO 6FLHQFH ([SHULPHQW RQH XVHG ILQLVKLQJ SLJV NJf WR GHWHUPLQH WKH SRWHQF\ RI YLWDPLQ ( FRPSRXQGV 3LJV ZHUH GLYLGHG DPRQJ ILYH QXWULWLRQDOO\ DGHTXDWH GLHWV VXSSOHPHQWHG ZLWK '/DWRFRSKHURO '/DWRFRSKHU\O DFHWDWH 'D WRFRSKHURO 'DWRFRSKHU\ DFHWDWH RU QR YLWDPLQ ( %ORRG DQG WLVVXH VDPSOHV ZHUH FROOHFWHG 9LWDPLQ ( IRUPV LQFUHDVHG 3 f VHUXP DWRFRSKHURO FRQFHQWUDWLRQV E\ G RI WKH IHHGLQJ SHULRG 6HUXP WRFRSKHURO LQ SLJV IHG DFHWDWH IRUPV UHPDLQHG HOHYDWHG WKURXJK RXW WKH VWXG\ VHUXP FRQFHQWUDWLRQV GHFOLQHG 3 f LQ SLJV IHG DOFRKRO IRUPV 'DWRFRSKHU\ DFHWDWH UHVXOWHG LQ KLJKHVW VHUXP DQG WLVVXH WRFRSKHURO 7KH SRWHQF\ RI 'DFHWDWH IRUP ZDV JUHDWHU IRU VZLQH WKDQ WKDW SUHGLFWHG IURP ELRDVVD\V ZLWK WKH UDW ([SHULPHQW WZR HYDOXDWHG H[FHVVLYH GLHWDU\ YLWDPLQ $ RQ YLWDPLQ ( VWDWXV DQG SHUIRUPDQFH RI JURZLQJILQLVKLQJ SLJV YLLL

PAGE 9

(LJKW\IRXU SLJV ZHUH IHG FRUQVR\EHDQ PHDO EDVHG GLHWV VXSSOHPHQWHG ZLWK '/DWRFRSKHU\ DFHWDWH WR SURYLGH RU ,8 RI YLWDPLQ (NJ DQG ZLWK UHWLQ\O DFHWDWH WR SURYLGH RU ,8 RI YLWDPLQ $NJ RI GLHW 6HUXP DQG WLVVXH WRFRSKHURO LQFUHDVHG 3 f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f DW G DJDLQ DW EUHHGLQJ G f DQG DW G SRVWEUHHGLQJ 5HSURGXFWLYH SHUIRUPDQFH ZDV QRW DIIHFWHG E\ WUHDWPHQW 6HUXP WRFRSKHURO LQFUHDVHG 3 f ZLWK ,8 RI YLWDPLQ ( +LJK ,8NJf GLHWDU\ YLWDPLQ ( LQFUHDVHG WRFRSKHURO OHYHO 3 f LQ DOO WLVVXHV H[FHSW DGLSRVH +LJK YLWDPLQ $ ,8f YLD LQMHFWLRQV KDG QR FRQVLVWHQW HIIHFW RQ UHSURGXFWLYH SHUIRUPDQFH RU RQ VHUXP RU WLVVXH FRQFHQWUDWLRQV RI DWRFRSKHURO RU UHWLQRO ,;

PAGE 10

&+$37(5 ,1752'8&7,21 9LWDPLQ ( ZDV GLVFRYHUHG LQ DV D PLVVLQJ QHHGHG GLHWDU\ IDFWRU %UDQGQHU 8OOUH\ 5DDFNH 0F'RZHOO f 9LWDPLQ ( ZDV LVRODWHG DV DOSKD WRFRSKHURO 7KH QDPH WRFRSKHURO PHDQV WR EULQJ IRUWK RIIVSULQJ 0F'RZHOO f *HRUJH 0 &DOKRXQ D SURIHVVRU RI *UHHN DW WKH 8QLYHUVLW\ RI &DOLIRUQLD %HUNHOH\ QDPHG WKH QHZ YLWDPLQ WRFRSKHURO LQ WRFRV IRU FKLOGELUWK SKHUR WR FRQIHU DQG RO IRU DOFRKROf (YDQV 8OOUH\ 5DDFNH f ,W ZDV UHFRJQL]HG LQ WKDW UHSURGXFWLYH IDLOXUH RFFXUUHG LQ UDWV FRQVXPLQJ GLHWV WKRXJKW WR EH QXWULWLRQDOO\ DGHTXDWH $Q XQNQRZQ GLHWDU\ IDFWRU WKHQ FDOOHG ; DQG ODWHU GHWHUPLQHG WR EH YLWDPLQ ( ZDV GHILFLHQW ZKLFK UHVXOWHG LQ IHWDO GHDWK DQG HPEU\R UHVRUSWLRQ LQ WKH ODERUDWRU\ UDW (YDQV 0DVRQ 'LSORFN 0F'RZHOO f (VWUXV DQG PDWLQJ ZHUH QRUPDO EXW IHWXVHV GLHG DQG ZHUH UHVRUEHG XQOHVV WKH GLHW ZDV VXSSOHPHQWHG ZLWK VPDOO DPRXQWV RI ZKHDW JHUP GULHG DOIDOID OHDYHV RU IUHVK OHWWXFH ZKLFK FRQWDLQHG WKH GHILFLHQW YLWDPLQ ( (YDQV 0DVRQ 0F'RZHOO f 'HJHQHUDWLRQ RI WKH JHUPLQDO HSLWKHOLXP LQ PDOH UDWV ZDV SUHYHQWHG E\

PAGE 11

VXSSOHPHQWV RI IUHVK OHWWXFH 0DVRQ f 2WKHU DQLPDO VSHFLHV FDWWOH VKHHS PLQN DQG FKLFNHQVf ZHUH DEOH WR UHSURGXFH ZLWKRXW GLHWDU\ YLWDPLQ ( EXW LQ HDFK FDVH WKHLU RIIVSULQJ GLHG SUHPDWXUHO\ %UDQGQHU f 9LWDPLQ ( EHFDPH NQRZQ DV WKH IHUWLOLW\ YLWDPLQ 0DQ\ VWXGLHV ZHUH GRQH WR GHWHUPLQH LI YLWDPLQ ( DIIHFWHG UHSURGXFWLRQ LQ KXPDQV ,Q PRVW FDVHV YLWDPLQ ( KDG OLWWOH RU QR HIIHFW 0F'RZHOO f 9LWDPLQ ( GHILFLHQF\ LQ VZLQH UHVXOWV LQ UHGXFHG UHSURGXFWLYH HIILFLHQF\ ORFRPRWRU LQFRRUGLQDWLRQ PXVFXODU DQG KHSDWLF QHFURVLV ILEULQRLG GHJHQHUDWLRQ RI EORRG YHVVHO ZDOOV DQG PXVFXODU G\VWURSK\ 0F'RZHOO f 9LWDPLQ ( LV D K\GURSKRELF SHUR[\O UDGLFDOWUDSSLQJ FKDLQEUHDNLQJ DQWLR[LGDQW IRXQG LQ WKH OLSLG IUDFWLRQ RI OLYLQJ RUJDQLVPV ,WV SULQFLSDO IXQFWLRQ LV WR SURWHFW WKH OLSLG PDWHULDO RI DQ RUJDQLVP IURP R[LGDWLRQ 0DFKOLQ %XUWRQ HW DO 'LSORFN 0F'RZHOO &RHOKR f /LSLG SHUR[LGDWLRQ RI PHPEUDQHV RI FHOOV DQG FHOOXODU FRQVWLWXHQWV FDQ EH YHU\ GDPDJLQJ 'DPDJH PD\ EH DV VLPSOH DV EUHDNLQJ D PHPEUDQH DQG DOORZLQJ OHDNDJH RI FRQWHQWV RU DV FRPSOH[ DV EUHDNLQJ D PHPEUDQH FRQWDLQLQJ GHVWUXFWLYH HQ]\PH V\VWHPV +HPRO\VLV RI UHG EORRG FHOOV LV DQ H[DPSOH RI UHODWLYHO\ VLPSOH PHPEUDQH EUHDNDJH 0HPEUDQH GDPDJH WR O\VRVRPHV FDQ EH SDUWLFXODUO\ GHYDVWDWLQJ /\VRVRPHV DUH VRPHWLPHV FDOOHG WKH VXLFLGH EDJV RI WKH FHOO DQG ZKHQ WKHLU PHPEUDQHV DUH EURNHQ WKH\ UHOHDVH

PAGE 12

HQ]\PHV WKDW K\GURO\]H WLVVXH FRQVWLWXHQWV DQG PDJQLI\ WLVVXH GDPDJH 7DSSHO f 'DPDJH WR WKH PHPEUDQH RI VXFK RWKHU FHOOXODU FRPSRQHQWV DV PLWRFKRQGULD DQG PLFURVRPHV ZKLFK FRQWDLQ DQG b XQVDWXUDWHG OLSLG UHVSHFWLYHO\ KDYH SURIRXQG HIIHFWV ,Q ERWK PLFURVRPHV DQG PLWRFKRQGULD YLWDPLQ ( LV WKH RQO\ NQRZQ OLSLG DQWLR[LGDQW 7DSSHO f 6HOHQLXP 6Hf LV D WUDFH PLQHUDO WKDW LV NQRZQ WR VSDUH VRPH RI WKH UHTXLUHPHQW IRU YLWDPLQ ( 6HOHQLXP LV D FRPSRQHQW RI WKH HQ]\PH JOXWDWKLRQH SHUR[LGDVH ZKLFK LV D VHOHQRSURWHLQ FRQWDLQLQJ IRXU DWRPV RI VHOHQLXP SHU PROHFXOH RI SURWHLQ 6FRWW 'UDSHU f *OXWDWKLRQH LV D K\GURJHQ GRQRU 9LWDPLQ ( IXQFWLRQV DV D IDW VROXEOH DQWLR[LGDQW DQG VHOHQLXP IXQFWLRQV DV D ZDWHU VROXEOH DQWLR[LGDQW &XQKD f 9LWDPLQ ( LV WKH ILUVW OLQH RI GHIHQVH DJDLQVW SHUR[LGDWLRQ RI IDWV LQ FHOOV ,I SHUR[LGHV DUH IRUPHG VHOHQLXP WKURXJK WKH HQ]\PH JOXWDWKLRQH SHUR[LGDVH GHVWUR\V WKH SHUR[LGHV EHIRUH WLVVXH GDPDJH FDQ RFFXU 7KXV VHOHQLXP LV FRQVLGHUHG WKH VHFRQG OLQH RI GHIHQVH 'LSORFN 0F'RZHOO f DQG DV D UHVXOW ERWK VHOHQLXP DQG YLWDPLQ ( DUH FDSDEOH RI SUHYHQWLQJ VRPH RI WKH VDPH QXWULWLRQDO GLVHDVHV 0F'RZHOO f 9LWDPLQ ( FDQ DOVR UHGXFH WKH VHOHQLXP UHTXLUHPHQW E\ LQKLELWLQJ SURGXFWLRQ RI SHUR[LGHV 3LJV H[KLELWLQJ FOLQLFDO YLWDPLQ ( DQG 6H GHILFLHQF\ VLJQV KDYH D SDOH ZKLWH GLVFRORUDWLRQ RI WKH VNHOHWDO

PAGE 13

PXVFOH DQ HQODUJHG IULDEOH KHDUW DVVRFLDWHG ZLWK K\GURSHULFDUGLXP DQG VRPHWLPHV LQWHVWLQDO HGHPD DQG KHSDWRVLV GLHWWLFD 0DKDQ DQG 0R[RQ f 7KHUH DUH PDQ\ IDFWRUV WKDW DIIHFW WKH ELRDYDLODELOLW\ RI YLWDPLQ ( 7KHVH LQFOXGH WKH IRUP RI YLWDPLQ ( FRPSRXQG SRWHQF\ RI FRPSRXQG VWDELOLW\ DEVRUSWLRQ RWKHU IDW VROXEOH YLWDPLQV HJ UHWLQROf PLQHUDO LQWHUDFWLRQV DQG XQVDWXUDWHG IDW %LRDYDLODELOLW\ LV GHILQHG DV WKH SHUFHQWDJH RI D GUXJ RU QXWULHQW LQ WKLV FDVH YLWDPLQ (f WKDW HQWHUV WKH V\VWHPLF FLUFXODWLRQ DIWHU DGPLQLVWUDWLRQ DQG WKH UDWH RI HQWU\ LQWR WKH JHQHUDO FLUFXODWLRQ IRU GLVWULEXWLRQ WKURXJKRXW WKH ERG\ DV ZHOO DV WLVVXH DFFXPXODWLRQ .RFK:HVHU 7KH $PHULFDQ +HULWDJH 'LFWLRQDU\ f (LJKW IRUPV RI YLWDPLQ ( DUH NQRZQ WR RFFXU LQ QDWXUH IRXU RI ZKLFK DUH UHIHUUHG WR DV WRFRSKHUROV DQG IRXU DV WRFRWULHQROV 7KH\ KDYH EHHQ JLYHQ *UHHN OHWWHU QDPHV WR GLVWLQJXLVK WKHP IURP RQH DQRWKHU 'LSORFN 15& f 7KH FRPSRXQGV GLIIHU LQ WKH SODFHPHQW RI PHWK\O JURXSV RQ WKH ULQJ DQG WKH GHJUHH RI VDWXUDWLRQ LQ WKH VLGH FKDLQ 0F'RZHOO f $OSKDWRFRSKHURO LV D \HOORZ RLO VROXEOH LQ FHUWDLQ RUJDQLF VROYHQWV ,W LV FRPPRQ SUDFWLFH WR DVVD\ RQO\ WKLV LVRPHU UDWKHU WKDQ DOO HLJKW FRPSRXQGV EHFDXVH DWRFRSKHURO LV WKH PRVW ELRORJLFDOO\ DFWLYH QDWXUDOO\ RFFXUULQJ YLWDPLQ ( VRXUFH 8OOUH\ f

PAGE 14

'/DWRFRSKHURO KDV D SRWHQF\ RI ,8PJ DQG LWV DFHWDWH '/DWRFRSKHU\ DFHWDWHf KDV D SRWHQF\ RI ,8PJ DV GHWHUPLQHG E\ ELRDVVD\V ZLWK UDWV $FWLYLW\ RI QDWXUDOO\ RFFXUULQJ DWRFRSKHURO 'DWRFRSKHURO DOVR FDOOHG 555 WRFRSKHUROf LV ,8PJ DQG RI LWV DFHWDWH ,8PJ 'DWRFRSKHURO LV WKH PRVW ELRORJLFDOO\ DFWLYH IRUP ,8 SHU XQLW RI ZHLJKW 15& f /RVV RI YLWDPLQ ( SRWHQF\ RFFXUV LQ PL[HG IHHG IURP D QXPEHU RI IDFWRUV 7KH QDWXUDOO\ RFFXUULQJ WRFRSKHUROV KDYH UHODWLYHO\ SRRU VWDELOLW\ GXULQJ SURFHVVLQJ JULQGLQJ SHOOHWLQJ DQG VWRULQJ DW KLJK WHPSHUDWXUHV RU XQGHU PRLVW FRQGLWLRQV 9LWDPLQ ( ZLOO DOVR UHDGLO\ LQWHUDFW ZLWK RWKHU LQJUHGLHQWV LQ IHHG IRUPXODWLRQV $GDPV 15& f 0RUH SLJV DUH EHLQJ UDLVHG LQ FRQILQHPHQW ZLWKRXW DFFHVV WR SDVWXUH ZKLFK LV DQ H[FHOOHQW VRXUFH RI YLWDPLQ ( +HDWLQJ DQG SHOOHWLQJ IHHG JUDLQV ORZHU WKHLU YLWDPLQ ( YDOXHV 7KH XVH RI KLJK PRLVWXUH JUDLQ LQFUHDVHV WKH QHHG IRU YLWDPLQ ( VXSSOHPHQWDWLRQ GXH WR WKH GHVWUXFWLRQ RI WKH YLWDPLQ )HHGV IRUPXODWHG ZLWK IDWV FRQWDLQLQJ KLJK TXDQWLWLHV RI XQVDWXUDWHG IDWW\ DFLGV DUH VXVFHSWLEOH WR UDQFLGLW\ ZKLFK GHVWUR\V YLWDPLQ ( &XQKD f 0DOP HW DO f UHSRUWHG WKDW GLHWV KLJK LQ SRO\XQVDWXUDWHG IDWW\ DFLGV LQFUHDVHG YLWDPLQ ( UHTXLUHPHQW DQG WKDW SLJV IHG D SRO\XQVDWXUDWHG IDWW\ DFLG ORZ YLWDPLQ ( GLHW WKURXJKRXW WKH SRVWZHDQLQJ SHULRG UHVXOWHG LQ VRPH GHJUHH RI UHG EORRG FHOO GHVWUXFWLRQ

PAGE 15

7KH DOFRKRO IRUP DWRFRSKHURO LV HDVLO\ GHVWUR\HG E\ R[LGDWLRQ 2[LGDWLYH GHVWUXFWLRQ RI DWRFRSKHURO LV DFFHOHUDWHG E\ KHDW OLJKW PRLVWXUH XQVDWXUDWHG IDWV VXOIDWHV QLWUDWHV DQG PROGV DQG LQ GLHWV FRQWDLQLQJ LQFUHDVHG OHYHOV RI FRSSHU LURQ ]LQF DQG PDQJDQHVH 8OOUH\ 0F'RZHOO 'RYH DQG (ZDQ 0DKDQ 7KRPSVRQ f $ PRUH VWDEOH VRXUFH RI YLWDPLQ ( LV DWRFRSKHU\O DFHWDWH $OSKDWRFRSKHU\ DFHWDWH LV FKHPLFDOO\ V\QWKHVL]HG E\ HVWHULILFDWLRQ RI DWRFRSKHURO ZLWK DFHWLF DFLG '/DWRFRSKHU\ DFHWDWH LV WKH LQWHUQDWLRQDO VWDQGDUG IRU YLWDPLQ ( DFWLYLW\ 9LWDPLQ ( LV IDW VROXEOH DQG DV VXFK LWV DEVRUSWLRQ LV DVVRFLDWHG ZLWK WKDW RI OLSLGV 9LWDPLQ ( LV DEVRUEHG LQ WKH DOFRKRO IRUP 9LWDPLQ ( DFHWDWH LV K\GURO\]HG WR WKH DOFRKRO IRUP LQ WKH VPDOO LQWHVWLQH SULRU WR DEVRUSWLRQ 'URSOHWV RI WULJO\FHULGHV DUH GHJUDGHG E\ OLSDVH DQG ELOH LQWR PRQRJO\FHULGHV DQG IUHH IDWW\ DFLGV ZKLFK IRUP LQWR PLFHOOHV 0LFHOOHV FRQWDLQ WKH OLSLG FRPSRQHQWV LQFOXGLQJ WKH IDW VROXEOH YLWDPLQV 9LWDPLQV DUH DEVRUEHG ZLWK WKH IDWW\ DFLGV DQG PRQRJO\FHULGHV 7ULJO\FHULGHV DUH UHIRUPHG LQ WKH LQWHVWLQDO FHOO DQG SDFNDJHG LQWR FK\ORPLFURQV &K\ORPLFURQV DUH DEVRUEHG LQWR WKH ODFWHDO GXFWV DQG FDUULHG LQWR WKH O\PSKDWLF V\VWHP XQWLO WKH\ HQWHU WKH JHQHUDO FLUFXODWLRQ DQG DUH GLVWULEXWHG WR YDULRXV WLVVXHV )DFWRUV LQWHUIHULQJ ZLWK GLJHVWLRQ DQG DEVRUSWLRQ RI OLSLG DIIHFW WKH ELRDYDLODELOLW\ RI YLWDPLQ (

PAGE 16

&RPSHWLWLRQ IRU DEVRUSWLRQ VLWHV LQ WKH VPDOO LQWHVWLQHV DPRQJ WKH IDW VROXEOH YLWDPLQV PD\ DIIHFW ELRDYDLODELOLW\ RI YLWDPLQ ( 9LWDPLQ $ UHWLQROf PD\ LQWHUIHUH ZLWK ERWK DEVRUSWLRQ DQG EORRG FRQFHQWUDWLRQV RI YLWDPLQ ( 7KLV KDV EHHQ GHPRQVWUDWHG LQ FKLFNV 6NODQ DQG 'RQRJKXH $EDZL DQG 6XOOLYDQ f DQG UDWV %ODNHO\ HW DO f 7KLV HIIHFW DSSHDUV WR EH GXH WR LQFUHDVHG R[LGDWLRQ RI YLWDPLQ ( SULRU WR WKH GLJHVWD UHDFKLQJ WKH GXRGHQXP 7KLV ZRXOG UHVXOW LQ YLWDPLQ ( FRQFHQWUDWLRQ EHLQJ ORZHU DW WKH PDMRU DEVRUSWLRQ VLWHV LQ WKH XSSHU VPDOO LQWHVWLQH 6NODQ DQG 'RQRJKXH f ,Q WKLV FDVH YLWDPLQ ( LV R[LGL]HG DW WKH H[SHQVH RI YLWDPLQ $ (UGPDQ HW DO f UHSRUWHG WKDW YLWDPLQ ( PD\ SURWHFW YLWDPLQ $ IURP R[LGDWLRQ LQ WKH JDVWURLQWHVWLQDO WUDFW DQG ZLWKLQ FHOO PHPEUDQHV 5HSRUWV WKDW YLWDPLQ $ WR[LFLW\ LQ FKLFNV KDV EHHQ FRPSOHWHO\ UHYHUVHG ZLWK KLJK GLHWDU\ YLWDPLQ ( VXSSOHPHQWDWLRQ $UQULFK DQG $UWKXU f DOVR LQGLFDWH DQ DGGLWLRQDO ORVV RI YLWDPLQ ( UHVXOWLQJ LQ DQ LQFUHDVHG QHHG
PAGE 17

JHVWDWLQJ JLOWV DQG VRZV ([WUD YLWDPLQ $ JLYHQ MXVW EHIRUH GXULQJ DQG VKRUWO\ DIWHU EUHHGLQJ KDV EHHQ UHSRUWHG WR LPSURYH UHSURGXFWLYH SHUIRUPDQFH LQ EUHHGLQJ VZLQH %ULHI DQG &KHZ &RIIH\ DQG %ULWW f 7KH HOHYDWLRQ RI PDWHUQDO SODVPD YLWDPLQ $ LV EHOLHYHG WR LPSURYH HPEU\RQLF VXUYLYDO 15& f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

PAGE 18

&+$37(5 327(1&< 2) 9$5,286 9,7$0,1 ( &203281'6 )25 ),1,6+,1* 6:,1( ,QWURGXFWLRQ 9LWDPLQ ( LV DQ HVVHQWLDO QXWULHQW IRU QRUPDO JURZWK KHDOWK DQG UHSURGXFWLRQ LQ VZLQH 9LWDPLQ ( UHTXLUHPHQW IRU VZLQH UDQJHV EHWZHHQ DQG ,8NJ RI GLHW 15& f 6ZLQH GLHWV FRQVLVWLQJ PDLQO\ RI FRUQ DQG VR\EHDQ PHDO XVXDOO\ GR QRW FRQWDLQ DGHTXDWH DPRXQWV RI YLWDPLQ ( QHHGHG WR PHHW WKH SLJnV UHTXLUHPHQW 15& f ,Q DGGLWLRQ WKH VWDELOLW\ RI DOO QDWXUDOO\ RFFXUULQJ YLWDPLQ ( IRUPV DUH YHU\ SRRU LQ PL[HG IHHG 8OOUH\ 'RYH DQG (ZDQ +LGLURJORX HW DO f 7KHUHIRUH VXSSOHPHQWDWLRQ RI VZLQH GLHWV ZLWK D UHDGLO\ DYDLODEOH IRUP RI YLWDPLQ ( DVVXUHV WKDW VZLQH ZLOO UHFHLYH WKH FRUUHFW DPRXQW IRU RSWLPXP SHUIRUPDQFH (LJKW IRUPV RI YLWDPLQ ( RFFXU LQ QDWXUH WRFRSKHUROV WRFRWULHQROVf 'DWRFRSKHURO KDV WKH JUHDWHVW ELRORJLFDO DFWLYLW\ KLJKHVW ,8 SHU XQLW ZHLJKW 15& f EXW DFHWDWH DQG VXFFLQDWH IRUPV DUH PRUH VWDEOH (UGPDQ HW DO f 7KLV H[SHULPHQW HYDOXDWHG WKH UHODWLYH ELRSRWHQFLHV RI IRXU IRUPV RI YLWDPLQ ( '/D WRFRSKHU\O DFHWDWH 'DWRFRSKHU\ DFHWDWH '/DWRFRSKHURO DQG 'DWRFRSKHUROf ZKHQ VXSSOHPHQWHG LQ WKH GLHW RI

PAGE 19

ILQLVKLQJ VZLQH 7KH FRQFHQWUDWLRQ RI DWRFRSKHURO LQ EORRG VHUXP DQG WLVVXH ZDV XVHG DV DQ LQGLFDWRU RI SRWHQF\ ([SHULPHQWDO 3URFHGXUHV )RUW\ FURVVEUHG ILQLVKLQJ SLJV EDUURZV DQG JLOWV ZLWK DQ DYHUDJH LQLWLDO ZHLJKW RI NJ ZHUH UDQGRPO\ DVVLJQHG E\ VH[ WR LQGLYLGXDO SHQV 7UHDWPHQWV ZHUH UDQGRPO\ DVVLJQHG WR WKH SHQV VXFK WKDW HDFK WUHDWPHQW FRQVLVWHG RI SLJV EDUURZV JLOWVf 7UHDWPHQWV FRQVLVWHG RI WKH IROORZLQJ VXSSOHPHQWHG YLWDPLQ ( IRUPV '/ DWRFRSKHURO '/DWRFRSKHU\O DFHWDWH 'DWRFRSKHURO DQG 'DWRFRSKHU\O DFHWDWH $ QHJDWLYH FRQWURO ZKLFK UHFHLYHG QR VXSSOHPHQWDO YLWDPLQ ( ZDV DOVR LQFOXGHG WR JLYH D ILIWK WUHDWPHQW 9LWDPLQ ( IRUPV XVHG ZHUH SXUH IRUPV VXSSOLHG LQ DQ XQSURWHFWHG RLO VROXWLRQ 6LJPD &KHPLFDO &R 6W /RXLV 02f 6XSSOHPHQWDO YLWDPLQ ( ZDV DGGHG WR WKH GLHWV VXFK WKDW SLJV FRQVXPLQJ NJ RI IHHG ZRXOG UHFHLYHG ,8 SHU GD\ 3LJV ZHUH IHG D FRUQVR\EHDQ PHDO ILQLVKLQJ GLHW IRUPXODWHG ZLWK D PRGLILHG YLWDPLQ SUHPL[ H[FOXVLYH RI YLWDPLQ (f DQG SSP RI DGGHG VHOHQLXP 'LHWV ZHUH RWKHUZLVH IRUPXODWHG IROORZLQJ 15& f JXLGHOLQHV &RPSRVLWLRQ RI WKH FRUQVR\EHDQ PHDO EDVDO GLHW LV JLYHQ LQ 7DEOH 7KH SLJV ZHUH IHG WKH ILQLVKHU GLHW IRU GD\V 3ULRU WR WKH VWDUW RI WKLV WULDO DOO SLJV ZHUH IHG D GLHW WKDW FRQWDLQHG ,8 RI YLWDPLQ (NJ RI GLHW )HHG DQG ZDWHU ZHUH DYDLODEOH DG OLELWXP WKURXJKRXW WKH H[SHULPHQW

PAGE 20

7DEOH &RPSRVLWLRQ RI ILQLVKHU GLHW ,QJUHGLHQW b DV IHG *URXQG FRUQ 6R\EHDQ PHDO bf '\QDIRV *URXQG OLPHVWRQH 6DOW 7UDFH PLQHUDO 9LWDPLQ PL[n 6H SUHPL[F f3URYLGHG SSP ]LQF SSP LURQ SSP PDQJDQHVH SSP FRSSHU DQG SSP LRGLQH E3URYLGHG PJ ULERIODYLQ PJ QLDFLQ PJ SDQWRWKHQLF DFLG PJ FKROLQH FKORULGH XJ YLWDPLQ % PJ YLWDPLQ ,8 YLWDPLQ $ DQG ,8 YLWDPLQ SHU NJ RI GLHW f3URYLGHG SSP VHOHQLXP

PAGE 21

3LJV ZHUH KRXVHG LQ DQ RSHQVLGHG EXLOGLQJ ZLWK VROLG FRQFUHWH IORRUV ,QGLYLGXDO SLJ ZHLJKWV DQG IHHG FRQVXPSWLRQ ZHUH UHFRUGHG ELZHHNO\ 7KH WULDO ZDV FDUULHG RXW LQ WKH VSULQJ $SULO0D\f RI WKH \HDU 3LJV ZHUH PDQDJHG DFFRUGLQJ WR DFFHSWDEOH PDQDJHPHQW SUDFWLFHV WKURXJKRXW WKH H[SHULPHQW 3URWRFRO IRU DQLPDO FDUH KDG EHHQ DSSURYHG E\ WKH 8QLYHUVLW\ $QLPDO 8VH &RPPLWWHH %ORRG VDPSOHV ZHUH FROOHFWHG E\ MXJXODU YHLQ SXQFWXUH IURP HDFK SLJ RQ G DQG RI WKH IHHGLQJ SHULRG %ORRG VDPSOHV ZHUH FHQWULIXJHG DIWHU FROOHFWLRQ DQG VHUXP ZDV IUR]HQ DQG VWRUHG DW r& XQWLO DQDO\]HG )HHG VDPSOHV ZHUH WDNHQ IURP WKH IHHGHUV RQ G DQG IUR]HQ DQG VWRUHG DW r& XQWLO DQDO\]HG IRU D WRFRSKHURO FRQFHQWUDWLRQ 2Q G WKH EDUURZV ZHUH VODXJKWHUHG XVLQJ DFFHSWHG VODXJKWHU SURFHGXUHV DW WKH 8QLYHUVLW\ RI )ORULGD PHDWV ODERUDWRU\ DQG WLVVXH VDPSOHV FROOHFWHG 7LVVXH VDPSOHV LQFOXGHG OLYHU PXVFOH UKRPERLGHXV DQG VHPLPHPEUDQRVXVf EDFN IDW WK ULE DUHDf DQG OHDI IDW 7LVVXH VDPSOHV ZHUH IUR]HQ IROORZLQJ FROOHFWLRQ DQG VWRUHG DW r& XQWLO DQDO\]HG 3URFHGXUHV XVHG IRU WKH H[WUDFWLRQ DQG GHWHUPLQDWLRQ RI DWRFRSKHURO LQ EORRG VHUXP ZHUH DV SUHYLRXVO\ GHVFULEHG 1MHUX HW DO f 3URFHGXUHV ZHUH VLPLODU WR WKRVH XVHG E\ 0F0XUUD\ DQG %ODQFKIORZHU DEf H[FHSW LQ RXU VWXG\ SURSDQRO ZDV XVHG LQ WKH VHUXP H[WUDFWLRQ LQVWHDG RI HWKDQRO 0RVW RI WKH YLWDPLQ ( DFWLYLW\ LQ VHUXP DQG WLVVXH

PAGE 22

ZDV DVVXPHG WR EH DWRFRSKHURO 8OOUH\ f ([WUDFWLRQ RI YLWDPLQ ( IURP WLVVXHV DQG IHHG ZDV GRQH XVLQJ D SURFHGXUH RXWOLQHG E\ &KXQJ HW DO f 7KLV SURFHGXUH ZDV D PRGLILFDWLRQ RI WKDW RI 0F0XUUD\ DQG %ODQFKIORZHU Ef DQG +DWDP DQG .D\GHQ f $OSKDWRFRSKHURO FRQFHQWUDWLRQ ZDV GHWHUPLQHG XVLQJ XO RI WKH UHFRQVWLWXWHG VDPSOH VHUXP WLVVXH RU IHHGf LQMHFWHG RQWR D /L&KURVRUE 6, FROXPQ +LEDU )HUWLJVDXOH 57 SUHSDFNHG FROXPQ 57 ( 0HUFN 'DUPVWDGW *HUPDQ\f PP [ PP ,' DQG XVLQJ D 3HUNLQ (OPHU WHUPLQDO 3HUNLQ(OPHU &RUS $QDO\WLFDO ,QVWUXPHQWV 1RUZDON &7f D 3HUNLQ (OPHU ,66 DXWR VDPSOHU DQG D 3HUNLQ (OPHU 6HULHV /LTXLG FKURPDWRJUDSK SXPS 7KH PRELOH SKDVH FRQVLVWHG RI +3/& JUDGH LVRRFWDQH WHWUDK\GURIXUDQ DQG DFHWLF DFLG 7KH GHWHFWRU ZDV D 3HUNLQ (OPHU /6 )OXRUHVFHQFH 6SHFWURPHWHU ZLWK DQ H[FLWDWLRQ ZDYHOHQJWK RI QP DQG DQ HPLVVLRQ ZDYHOHQJWK RI QP 'DWD ZHUH FROOHFWHG E\ D 3HUNLQ (OPHU /&, /DERUDWRU\ &RPSXWLQJ ,QWHJUDWRU )ORZ UDWH ZDV POPLQ 7KH UHWHQWLRQ WLPH RI DWRFRSKHURO ZDV PLQXWHV $OSKDWRFRSKHURO (DVWPDQ .RGDN &RPSDQ\ 5RFKHVWHU 1
PAGE 23

3RWHQFLHV RI WKH YDULRXV YLWDPLQ ( FRPSRXQGV ZHUH GHWHUPLQHG E\ FRPSDULQJ DUHDV XQGHU WKH WLPH FXUYH $8&f ZLWKLQ WKH VHUXP IHHG DQG WLVVXH VDPSOHV 6HUXP DQG WLVVXH FRQFHQWUDWLRQV ZHUH DGMXVWHG EDVHG RQ DFWXDO IHHG LQWDNH DQG DWRFRSKHURO OHYHOV LQ WKH IHHG $OSKDWRFRSKHURO FRQFHQWUDWLRQV UHSRUWHG ZHUH DGMXVWHG WR D FRQVWDQW IHHG D WRFRSKHURO FRQFHQWUDWLRQ G IHHG OHYHOf 6HUXP DQG WLVVXH PHDQV ZHUH DQDO\]HG XVLQJ WKH JHQHUDO OLQHDU PRGHO SURFHGXUH 6$6 f $QDO\VLV RI YDULDQFH FRPSDUHG WUHDWPHQW GLIIHUHQFHV LQ VHUXP DQG WLVVXH WRFRSKHURO FRQFHQWUDWLRQV $QDO\VLV RI FRYDULDQFH ZDV DOVR DSSOLHG WR WKH VHUXP GDWD XVLQJ EDVHOLQH G VHUXP WRFRSKHUROf GDWD DV D FRYDULDWH 7UHDWPHQW PHDQV ZHUH FRPSDUHG XVLQJ WKH OHDVW VLJQLILFDQW GLIIHUHQFH PXOWLSOH FRPSDULVRQ SURFHGXUH 5HVXOWV DQG 'LVFXVVLRQ *URZWK UDWH RI DOO SLJV ZDV JRRG RYHU WKH GXUDWLRQ RI WKH G VWXG\ 'DLO\ IHHG LQWDNH DQG IHHGWRJDLQ UDWLR ZHUH QRW DIIHFWHG 3 f E\ VXSSOHPHQWDWLRQ RI WKH YLWDPLQ ( VRXUFHV 7DEOH f KRZHYHU D VOLJKW LPSURYHPHQW LQ JURZWK UDWH 3 f ZDV REWDLQHG LQ SLJV IHG WKH 'D WRFRSKHURO IRUP FRPSDUHG WR WKH QHJDWLYH FRQWURO $VJKDU HW DO f UHSRUWHG LPSURYHG JURZWK UDWHV LQ JURZLQJn ILQLVKLQJ SLJV IHG GLHWDU\ OHYHOV RI '/DWRFRSKHU\ DFHWDWH DW ,8NJ RI GLHW FRPSDUHG WR SLJV IHG ,8NJ RI GLHW ,Q FRQWUDVW &KXQJ HW DO f IRXQG QR GLIIHUHQFH LQ

PAGE 24

7DEOH 3HUIRUPDQFH RI ILQLVKLQJ SLJV IHG GLHWV FRQWDLQLQJ YDULRXV YLWDPLQ ( FRPSRXQGV ,WHP 9LWDPLQ ( VRXUFH '/D7$& 'D7$& '/D72+ 'D72+ 1HJ FRQWURO 1R RI SLJV $'* NJ E E EF F E $') NJ )* D'/D7$& '/DWRFRSKHU\O DFHWDWH 'D7$& 'D WRFRSKHU\O DFHWDWH '/D72+ '/DWRFRSKHURO 'D72+ 'DWRFRSKHURO EF0HDQV ZLWKLQ WKH VDPH URZ ZLWK D GLIIHUHQW VXSHUVFULSW GLIIHU VLJQLILFDQWO\ 3 f

PAGE 25

JURZWK SHUIRUPDQFH GXH WR YLWDPLQ ( VRXUFH HQFDSVXODWHG DWRFRSKHURO RU '/DWRFRSKHU\O DFHWDWHf RU OHYHO DQG ,8NJf LQ WULDOV ZLWK \RXQJ VWDUWLQJ VZLQH $OSKDWRFRSKHURO DQDO\VLV RI WKH GLHWV FRQWDLQLQJ WKH YLWDPLQ ( IRUPV DUH UHSRUWHG LQ 7DEOH 7KH YLWDPLQ ( IRUPV ZHUH LQFOXGHG LQ WKH GLHW VR WKDW SLJV ZRXOG FRQVXPH ,8 RI DGGHG YLWDPLQ (NJ RI IHHG ,8NJ WRWDOf +RZHYHU WKHUH ZDV FRQVLGHUDEOH YDULDWLRQ LQ DQDO\]HG OHYHOV DPRQJ WKH GLHWDU\ WUHDWPHQWV $OVR WKHUH ZDV VRPH YDULDWLRQ LQ IHHG FRQVXPSWLRQ DPRQJ WUHDWPHQWV 7KHUHIRUH GDWD UHSRUWHG ZHUH DGMXVWHG EDVHG RQ DQDO\VLV RI GLHWV DQG PHDQ WUHDWPHQW JURXS IHHG FRQVXPSWLRQ 7DEOH f 7KH LQGLFDWRU XVHG WR GHWHUPLQH ELRSRWHQF\ RI WKH YLWDPLQ ( FRPSRXQGV ZDV WKH FRQFHQWUDWLRQ RI DWRFRSKHURO LQ VHUXP DQG VHOHFWHG WLVVXHV %UDW]OHU HW DO f IRXQG WKDW SODVPD WRFRSKHURO FRQFHQWUDWLRQ UHIOHFWHG OHYHO RI WRFRSKHURO LQJHVWHG LQ WULDOV ZLWK \RXQJ SLJV IHG GLIIHUHQW FRQFHQWUDWLRQV RI WRFRSKHURO 7KH\ DOVR REVHUYHG LQFUHDVHV RI WRFRSKHURO LQ YDULRXV WLVVXHV 2WKHU UHVHDUFKHUV LQGLFDWHG WKDW DQ DQLPDOnV YLWDPLQ ( VWDWXV FDQ EH GHWHUPLQHG E\ PHDVXULQJ DWRFRSKHURO FRQFHQWUDWLRQ LQ VHUXP DQG YDULRXV WLVVXH DIWHU RUDO DGPLQLVWUDWLRQ %DNHU HW DO f ZLWK KXPDQV +LGLURJORX DQG 0F'RZHOO f ZLWK VKHHS DQG -HQVHQ HW DO f DQG $VJKDU HW DO f ZLWK SLJV 1XPHURXV VWXGLHV KDYH VKRZQ WKDW GLHWDU\ YLWDPLQ ( FRPSRXQGV DUH HIIHFWLYH LQ HOHYDWLQJ EORRG WRFRSKHURO

PAGE 26

7DEOH 9LWDPLQ ( DWRFRSKHUROf FRQFHQWUDWLRQV LQ IHHG 7UHDWPHQW YLWDPLQ ( VRXUFHf 6DPSOLQJ GD\D ,8NJ '/DWRFRSKHU\O DFHWDWH f f f f 'DWRFRSKHU\O DFHWDWH f f f f '/DWRFRSKHURO f f f f 'DWRFRSKHURO f f f f 1HJ FRQWURO D'D\ VDPSOHV ZHUH WDNHQ DIWHU VWDUW RI WULDO 6DPSOHV ZHUH WDNHQ GLUHFWO\ IURP IHHGHU WKHQ IUR]HQ XQWLO DQDO\]HG 1XPEHUV LQ SDUHQWKHVLV UHSUHVHQW DGMXVWHG OHYHOV DGMXVWHG WR D FRQVWDQW ,8NJ H[WUDSRODWHG IURP G OHYHOV

PAGE 27

FRQFHQWUDWLRQ DQG DOVR WKDW EORRG WRFRSKHURO FRQFHQWUDWLRQ LQFUHDVHG ZLWK LQFUHDVLQJ GLHWDU\ OHYHO RI YLWDPLQ ( +LGLURJORX HW DO -HQVHQ HW DO %HKUHQV DQG 0DGHUH $VJKDU HW DO &KXQJ HW DO f $V H[SHFWHG PHDQ VHUXP FRQFHQWUDWLRQV RI WRFRSKHURO DW G EDVHOLQHf ZHUH VLPLODU DFURVV DOO WUHDWPHQWV 7DEOH f $OO YLWDPLQ ( FRPSRXQGV IHG LQ WKLV H[SHULPHQW LQFUHDVHG 3 f VHUXP WRFRSKHURO FRQFHQWUDWLRQ 7KH LQFUHDVH LQ VHUXP WRFRSKHURO FRQFHQWUDWLRQ ZDV UDSLG 7KH LQFUHDVH VWDUWHG RQ G JUHZ IXUWKHU E\ G 3 f DQG SODWHDXHG E\ G +RUZLWW HW DO f QRWHG LQ D VWXG\ ZLWK KXPDQV WKDW VHUXP DWRFRSKHURO FRQFHQWUDWLRQV ZHUH LQFUHDVHG DW WR KU DIWHU LQJHVWLRQ RI YDULRXV YLWDPLQ ( IRUPV +RZDUG HW DO f ZLWK SLJV ZHDQHG DW G GHSOHWHG RI YLWDPLQ ( IRU WKH QH[W G DQG WKHQ IHG ,8 RI VXSSOHPHQWDO YLWDPLQ ( LQ WKH IRUP RI 'DWRFRSKHU\ DFHWDWH RU '/DWRFRSKHU\ DFHWDWH SHU NJ RI GLHW QRWHG D UDSLG LQFUHDVH LQ EORRG DWRFRSKHURO -HQVHQ HW DO f LQ D VWXG\ ZLWK SLJV G ROG DOVR REVHUYHG D UDSLG LQFUHDVH LQ VHUXP WRFRSKHURO FRQFHQWUDWLRQV DIWHU IHHGLQJ VXSSOHPHQWDO '/DWRFRSKHU\ DFHWDWH ,Q ERWK RI WKH DERYH VZLQH VWXGLHV WKH ILUVW EORRG VDPSOHV ZHUH WDNHQ G DIWHU WKH VWDUW RI WKH IHHGLQJ WULDO 6HUXP WRFRSKHURO FRQFHQWUDWLRQV RI SLJV IHG ERWK DFHWDWH IRUPV ZHUH PDLQWDLQHG EH\RQG G KRZHYHU OHYHOV GURSSHG VWHDGLO\ LQ SLJV IHG WKH DOFRKRO IRUPV DQG ZHUH

PAGE 28

7DEOH $GMXVWHG VHUXP YLWDPLQ ( WRFRSKHUROf FRQFHQWUDWLRQV LQ ILQLVKLQJ VZLQH IHG GLHWV VXSSOHPHQWHG ZLWK GLIIHUHQW YLWDPLQ ( FRPSRXQGV 'D\ 9LWDPLQ ( VRXUFH 1HJ FRQWURO '/D7$& 'D7$& '/D72+ 'D72+ LFM M P EF E OE E & & E & EF G & E & & G & E OF & G & E G G & & E G G G 1RWH (DFK PHDQ LV EDVHG RQ HLJKW REVHUYDWLRQV $GMXVWHG WR FRQVWDQW LQWDNH RI ,8NJ GLHW EDVHG RQ G IHHG DQDO\VHV 7DEOH f 'D\ VHUXP YDOXHV ZHUH QRW DGMXVWHG D'/D7$& '/DWRFRSKHU\ DFHWDWH 'D7$& 'D WRFRSKHU\O DFHWDWH '/D72+ '/DWRFRSKHURO 'D72+ 'DWRFRSKHURO NGF0HDQV ZLWKLQ WKH VDPH URZ ZLWK D GLIIHUHQW VXSHUVFULSW GLIIHU 3 f

PAGE 29

ORZHU 3 f RQ G DQG WKDQ WKH DFHWDWH IRUPV 7KLV GURS ZDV SUREDEO\ GXH WR SRRU VWDELOLW\ RI WKH DOFRKRO IRUPV LQ WKH IHHG 7DEOH f 'HJUDGDWLRQ RI YLWDPLQ ( RFFXUV WKURXJK R[LGDWLRQ DQG LV DFFHOHUDWHG E\ OLJKW DONDOL KHDW DQG WUDFH PLQHUDOV 8OOUH\ (UGPDQ HW DO 'RYH DQG (ZDQ +LGLURJORX HW DO f ,Q WKH DEVHQFH RI R[\JHQ WRFRSKHUROV DUH UHODWLYHO\ KHDW OLJKW DQG DONDOL VWDEOH 8OOUH\ f 6WDELOLW\ RI DWRFRSKHURO LV LQFUHDVHG E\ DF\ODWLRQ RI WKH FRPSRXQG 8OOUH\ f $FHWDWH IRUPV RI YLWDPLQ ( ZHUH IRXQG WR EH JXLWH VWDEOH LQ IHHG LQ RXU VWXG\ $FHWDWH IRUPV RI YLWDPLQ ( KDYH DOVR EHHQ QRWHG WR EH VWDEOH FRPSRXQGV E\ RWKHU UHVHDUFKHUV +DUULV DQG /XGZLJ DE 8OOUH\ 'RYH DQG (ZDQ &KXQJ HW DO f ,Q JHQHUDO VHUXP WRFRSKHURO FRQFHQWUDWLRQV REVHUYHG LQ WKH SUHVHQW VWXG\ ZHUH VLPLODU WR WKRVH RI RWKHU VWXGLHV LQ ZKLFK SLJV ZHUH IHG GLHWV FRQWDLQLQJ VLPLODU OHYHOV RI DGGHG YLWDPLQ ( -HQVHQ HW DO $VJKDU HW DO f :KLOH DOO YLWDPLQ ( IRUPV HYDOXDWHG UDSLGO\ LQFUHDVHG VHUXP WRFRSKHURO FRQFHQWUDWLRQV WKHUH ZDV VRPH HYLGHQFH RI D VOLJKW GLIIHUHQFH LQ WKH UDWH RI WKLV LQFUHDVH $YHUDJH VHUXP FRQFHQWUDWLRQ LQ SLJV IHG WKH '/ DFHWDWH IRUP ZDV QRW LQFUHDVHG 3 f XQWLO G ZKHUHDV VHUXP FRQFHQWUDWLRQV RI SLJV IHG WKH RWKHU FRPSRXQGV ZHUH LQFUHDVHG 3 f RQ G +RUZLWW HW DO f IRXQG WKDW 'WRFRSKHURO UDLVHG EORRG DWRFRSKHURO FRQFHQWUDWLRQV IDVWHU WKDQ WKH RU '/

PAGE 30

DWRFRSKHURO DFHWDWH IRUPV LQ UHVHDUFK GRQH ZLWK KXPDQV 'DWRFRSKHU\O DFHWDWH UHVXOWHG LQ KLJKHU VHUXP WRFRSKHURO OHYHOV WKDQ WKH '/ DFHWDWH IRUP 5HODWLYH WR '/DWRFRSKHU\O DFHWDWH WKH IRUP RI WKH VDPH FRPSRXQG KDG DQ DYHUDJH ELRSRWHQF\ RI b ,8 EDVLV 7DEOH f RU b ZHLJKW EDVLV ; f +RZDUG HW DO f GHWHUPLQHG D UHODWLYH ELRSRWHQF\ RI b ZHLJKW EDVLVf IRU 'FUWRFRSKHU\O DFHWDWH UHODWLYH WR WKH '/ IRUP RI WKH VDPH FRPSRXQG LQ WULDOV ZLWK JURZLQJ SLJV 7KXV LW ZRXOG DSSHDU WKDW WKH DFHWDWH IRUP KDV D KLJKHU ELRSRWHQF\ IRU VZLQH WKDQ WKDW GHWHUPLQHG IURP WKH WUDGLWLRQDO UDW IHWDO UHVRUSWLRQ ELRDVVD\V +RZHYHU $PHV f SUHVHQWHG HYLGHQFH WKDW WKH FRPPRQO\ DFFHSWHG FRQYHUVLRQ YDOXH RI PD\ EH WRR ORZ LQ WULDOV HYDOXDWLQJ WKH UHODWLYH ELRSRWHQF\ RI VHYHUDO YLWDPLQ ( FRPSRXQGV XVLQJ WKH UDW IHWDO UHVRUSWLRQ DVVD\ %RWK DOFRKRO IRUPV LQ RXU VWXG\ H[KLELWHG VLPLODU ELRSRWHQFLHV DQG WKHUH ZDV HYLGHQFH GXULQJ WKH HDUO\ SRUWLRQ RI WKH VWXG\ WKDW WKHVH IRUPV ZHUH VOLJKWO\ PRUH ELRSRWHQW WKDQ '/DWRFRSKHURO DFHWDWH 7DEOH f $OSKDWRFRSKHURO LQ WLVVXHV LQ JHQHUDO IROORZHG D VLPLODU SDWWHUQ WR WKDW REVHUYHG ZLWK VHUXP 7DEOH f 2YHUDOO SLJV IHG DQ\ RI WKH FRPSRXQGV KDG WLVVXH WRFRSKHURO FRQFHQWUDWLRQV KLJKHU WKDQ WKH QHJDWLYH FRQWURO 2WKHU UHVHDUFKHUV KDYH DOVR IRXQG WKDW DGGLQJ YLWDPLQ ( WR WKH GLHW LQFUHDVHG WLVVXH FRQFHQWUDWLRQV %UDW]OHU HW DO -HQVHQ HW DO $VJKDU HW DO f

PAGE 31

7DEOH 5HODWLYH ELRSRWHQF\ RI YLWDPLQ ( FRPSRXQGV bf &RPSRXQG GD\ GD\ GD\ GD\ GD\ GD\ $YJ 'D7$& '/D72+ 'D72+ 1RWH %DVHG RQ VHUXP YDOXHV '/DWRFRSKHU\O DFHWDWH ,8 EDVLV

PAGE 32

7DEOH $GMXVWHG WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV LQ ILQLVKLQJ VZLQH IHG GLHWV VXSSOHPHQWHG ZLWK GLIIHUHQW YLWDPLQ ( FRPSRXQGV 9LWDPLQ ( 6RXUFH 7LVVXH '/D7$& 'D7$& '/D72+ 'D72+ 1HJ &RQWURO /LYHU L L L a A E 0JJ & & G /HDI IDW %DFN )DW EF E FG & GF 5KRPERLGHXV E E FG r G 6HPLPHPEUDQRVXV FG E FG & G 1RWH (DFK PHDQ LV EDVHG RQ IRXU REVHUYDWLRQV $GMXVWHG WR FRQVWDQW LQWDNH RI ,8NJ GLHW EDVHG RQ G DQDO\VHV 7DEOH f 1HJDWLYH FRQWURO YDOXHV QRW DGMXVWHG D'/D7$& '/DWRFRSKHU\ DFHWDWH 'D7$& 'DWRFRSKHU\ DFHWDWH '/D72+ '/DWRFRSKHURO 'D72+ 'DWRFRSKHURO EFGF0HDQV ZLWKLQ WKH VDPH URZ ZLWK D GLIIHUHQW VXSHUVFULSW GLIIHU VLJQLILFDQWO\ 3 f

PAGE 33

*HQHUDOO\ DFHWDWH YLWDPLQ ( IRUPV UHVXOWHG LQ JUHDWHU WLVVXH WRFRSKHURO FRQFHQWUDWLRQV WKDQ DOFRKRO IRUPV DQG WKH GLIIHUHQFH ZDV VLJQLILFDQW 3 f LQ WKH OLYHU DQG UKRPERLGHXV 7KH JUHDWHU WLVVXH OHYHOV UHIOHFWHG WKH JUHDWHU VWDELOLW\ RI WKH DFHWDWH IRUPV LQ WKH IHHG RYHU WKH GXUDWLRQ RI WKH VWXG\ /LYHU WRFRSKHURO DYHUDJHG [JJ IRU WKH DFHWDWH IRUPV DQG cXJJ IRU WKH DOFRKRO IRUPV %HWZHHQ WKH WZR DFHWDWH IRUPV 'DWRFRSKHU\O DFHWDWH UHVXOWHG LQ WKH VDPH WLVVXH WRFRSKHURO FRQFHQWUDWLRQV DV WKH '/ IRUP ZLWK H[FHSWLRQ RI WKH VHPLPHPEUDQRVXV LQ ZKLFK WKH IRUP UHVXOWHG LQ KLJKHU FRQFHQWUDWLRQV 3 f 7LVVXH WRFRSKHURO FRQFHQWUDWLRQV ZHUH VLPLODU IRU WKH WZR DOFRKRO IRUPV /LYHU KDG WKH KLJKHVW WRFRSKHURO FRQFHQWUDWLRQ RI DOO WLVVXHV HYDOXDWHG 7KH OLYHU FRXOG EH DQ LQGLFDWRU RI GLHWDU\ YLWDPLQ ( VWDWXV RU UHIOHFW LPPHGLDWH VWDWXV -HQVHQ HW DO f LQGLFDWHG WKDW VHUXP DQG OLYHU D WRFRSKHURO FRQFHQWUDWLRQV UHIOHFWHG WKH VKRUW WHUP YLWDPLQ ( VWDWXV RI WKH SLJ DQG WKDW PXVFOH DQG IDW WLVVXH FRQFHQWUDWLRQV UHIOHFWHG WKH SLJnV ORQJWHUP YLWDPLQ ( VWDWXV 7KH FRQFHQWUDWLRQ RI WRFRSKHURO LQ DOO WLVVXHV RWKHU WKDQ WKH OLYHU ZDV VLPLODU 3 f LQ SLJV IHG WKH WZR DFHWDWH IRUPV ZLWK WKH H[FHSWLRQ RI VHPLPHPEUDQRVXV DV QRWHG DERYH -HQVHQ HW DO f LQ WULDOV ZLWK JURZLQJ SLJV DOVR QRWHG WKDW WKH OLYHU KDG WKH KLJKHVW

PAGE 34

FRQFHQWUDWLRQ RI WRFRSKHURO IROORZHG E\ DGLSRVH WLVVXH DQG VNHOHWDO PXVFOH $OVR $VJKDU HW DO f IHHGLQJ JURZLQJ SLJV '/DWRFRSKHU\O DFHWDWH DW ,8NJ RI GLHW REVHUYHG KLJKHU WRFRSKHURO FRQFHQWUDWLRQV LQ WKH OLYHU IROORZHG E\ WKH KHDUW OXQJ DQG NLGQH\ ,Q FRQFOXVLRQ DOO YLWDPLQ ( FRPSRXQGV HYDOXDWHG DOPRVW LPPHGLDWHO\ E\ G f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f LQGLYLGXDOO\ SHQQHG ZHUH GLYLGHG HTXDOO\ DPRQJ ILYH WUHDWPHQWV 7UHDWPHQWV FRQVLVWHG RI FRUQVR\EHDQ PHDO EDVHG GLHWV VXSSOHPHQWHG ZLWK '/DWRFRSKHURO '/DWRFRSKHU\O DFHWDWH 'DWRFRSKHURO RU 'DWRFRSKHU\ DFHWDWH $ WUHDWPHQW ZLWKRXW YLWDPLQ ( VXSSOHPHQWDWLRQ QHJDWLYH FRQWUROf VHUYHG DV WKH ILIWK WUHDWPHQW (DFK FRPSRXQG ZDV VXSSOHPHQWHG DW

PAGE 35

,8NJ RI GLHW %ORRG VDPSOHV ZHUH FROOHFWHG RQ G DQG 2Q G KDOI RI WKH SLJV ZHUH VODXJKWHUHG WR REWDLQ WLVVXH VDPSOHV )HHG VDPSOHV WDNHQ IURP IHHGHUV ZHUH FROOHFWHG RQ G DQG $OO YLWDPLQ ( IRUPV IHG LQFUHDVHG 3 f VHUXP DWRFRSKHURO FRQFHQWUDWLRQ E\ G DQG WKH FRQFHQWUDWLRQ SHDNHG E\ G 6HUXP WRFRSKHURO FRQFHQWUDWLRQV LQ SLJV IHG HLWKHU DFHWDWH IRUP UHPDLQHG HOHYDWHG EH\RQG G VHUXP FRQFHQWUDWLRQV VWHDGLO\ GHFOLQHG DQG ZHUH ORZHU 3 f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f WRFRSKHURO FRQFHQWUDWLRQV DV ZLWK VHUXP FRQFHQWUDWLRQV ZLWK WKH OLYHU KDYLQJ WKH KLJKHVW FRQFHQWUDWLRQ 7KH ELRSRWHQF\ RI 'DWRFRSKHU\O DFHWDWH IRU VZLQH DSSHDUV WR EH JUHDWHU WKDQ SUHGLFWHG IURP WUDGLWLRQDO ELRDVVD\V ZLWK UDWV

PAGE 36

&+$37(5 7+( ())(&7 2) (;&(66,9( ',(7$5< 9,7$0,1 $ 21 3(5)250$1&( $1' 9,7$0,1 ( 67$786 ,1 6:,1( )(' ',(76 9$5<,1* ,1 ',(7$5< 9,7$0,1 ( ,QWURGXFWLRQ %RWK YLWDPLQV $ DQG ( DUH IDW VROXEOH YLWDPLQV 7KHUH LV HYLGHQFH WKDW KLJK GLHWDU\ YLWDPLQ $ PD\ LQWHUIHUH ZLWK ERWK YLWDPLQ ( DEVRUSWLRQ DQG EORRG DWRFRSKHURO FRQFHQWUDWLRQ +LJK GLHWDU\ YLWDPLQ $ UHGXFHG DEVRUSWLRQ RI DWRFRSKHURO LQ WULDOV ZLWK FKLFNV 6NODQ DQG 'RQRJKXH f $EDZL DQG 6XOOLYDQ f QRWHG GHFUHDVHG SODVPD YLWDPLQ ( FRQFHQWUDWLRQ ZKHQ FKLFNV UHFHLYHG KLJK ,8NJf OHYHOV RI GLHWDU\ YLWDPLQ $ %ODNHO\ HW DO f DOVR UHSRUWHG WKDW KLJK GLHWDU\ YLWDPLQ $ WLPHV UHTXLUHPHQWf SOXV KLJK OHYHOV RI EHWD FDURWHQH PJNJ RI GLHWf UHGXFHG SODVPD YLWDPLQ ( FRQFHQWUDWLRQ E\ b LQ UDWV /LPLWHG UHVHDUFK LV DYDLODEOH RQ WKH HIIHFW RI GLHWDU\ YLWDPLQ $ RQ YLWDPLQ ( VWDWXV LQ SLJV 2XU VWXG\ ZDV GRQH WR HYDOXDWH WKH HIIHFW RI H[FHVVLYH GLHWDU\ YLWDPLQ $ RQ SHUIRUPDQFH DQG RQ VHUXP DQG WLVVXH FRQFHQWUDWLRQV RI D WRFRSKHURO RI JURZLQJILQLVKLQJ SLJV IHG GLHWV VXSSOHPHQWHG ZLWK YDU\LQJ OHYHOV RI YLWDPLQ (

PAGE 37

([SHULPHQWDO 3URFHGXUHV (LJKW\IRXU FURVVEUHG SLJV ZLWK DQ DYHUDJH LQLWLDO ZHLJKW RI NJ ZHUH GLYLGHG E\ VH[ ZHLJKW DQG OLWWHU RULJLQ LQWR SHQV RI WZR SLJV HDFK EDUURZ JLOWf (DFK SHQ ZDV DVVLJQHG WR RQH RI VL[ GLHWDU\ WUHDWPHQWV ZLWKLQ HDFK RI VHYHQ UHSOLFDWLRQV 7KH WUHDWPHQWV IRU WKH [ WULDO FRQVLVWHG RI D EDVDO FRUQVR\EHDQ PHDO GLHW VXSSOHPHQWHG ZLWK '/DWRFRSKHU\ DFHWDWH +RIIPDQQ/D 5RFKH ,QF 1XWOH\ 1-f DW OHYHOV RI RU ,8NJ DQG UHWLQ\O DFHWDWH +RIIPDQQ/D 5RFKH ,QF 1XWOH\ 1-f DW OHYHOV RI RU ,8NJ RI GLHW 3LJV ZHUH IHG D JURZHU GLHW 7DEOH f IRUPXODWHG WR PHHW 15& f UHTXLUHPHQWV H[FHSW IRU YLWDPLQV $ DQG (f XQWLO WKH\ UHDFKHG DQ DYHUDJH ERG\ ZHLJKW RI NJ DQG FRQWLQXHG RQ D ILQLVKHU GLHW 7DEOH f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

PAGE 38

7DEOH &RPSRVLWLRQ RI GLHWV bf ,QJUHGLHQW *URZHU )LQLVKHU *URXQG FRUQ 6R\EHDQ PHDO bf '\QDIRV *URXQG OLPHVWRQH 6DOW 7UDFH PLQHUDO SUHPL[D 9LWDPLQ SUHPL[ E F 6H SUHPL[G f3URYLGHG SSP ]LQF SSP LURQ SSP PDQJDQHVH SSP FRSSHU DQG SSP LRGLQH E3URYLGHG PJ ULERIODYLQ PJ QLDFLQ PJ SDQWRWKHQLF DFLG PJ FKROLQH FKORULGH XJ YLWDPLQ % PJ YLWDPLQ DQG ,8 YLWDPLQ SHU NJ RI GLHW f3URYLGHG PJ ULERIODYLQ PJ QLDFLQ PJ SDQWRWKHQLF DFLG PJ FKROLQH FKORULGH XJ YLWDPLQ % PJ YLWDPLQ DQG ,8 YLWDPLQ SHU NJ RI GLHW G3URYLGHG SSP VHOHQLXP

PAGE 39

GXULQJ ODWH VSULQJ DQG HDUO\ VXPPHU 0DUFK WKURXJK -XQHf 3LJV ZHUH PDQDJHG IROORZLQJ DFFHSWDEOH FDUH DQG PDQDJHPHQW SUDFWLFHV 3URWRFRO IRU DQLPDO FDUH KDG EHHQ DSSURYHG E\ WKH 8QLYHUVLW\ $QLPDO 8VH &RPPLWWHH %ORRG VDPSOHV ZHUH FROOHFWHG E\ MXJXODU YHQLSXQFWXUH IURP HDFK SLJ DW WKH VWDUW G f DQG RQ G DQG RI WKH IHHGLQJ SHULRG WKHUHDIWHU %ORRG VDPSOHV ZHUH VKLHOGHG IURP GLUHFW VXQOLJKW %ORRG VDPSOHV ZHUH FHQWULIXJHG DIWHU FROOHFWLRQ VHUXP KDUYHVWHG IUR]HQ DQG VWRUHG DW r& XQWLO DQDO\]HG 'XULQJ VWRUDJH EORRG VDPSOHV ZHUH FRYHUHG ZLWK IRLO WR DYRLG H[SRVXUH WR OLJKW 8SRQ WHUPLQDWLRQ RI WKH IHHGLQJ SKDVH RQH SLJ EDUURZf SHU SHQ ZDV VODXJKWHUHG IROORZLQJ DFFHSWHG VODXJKWHU SURFHGXUHV DW WKH 8QLYHUVLW\ RI )ORULGD PHDWV ODERUDWRU\ DQG WLVVXH VDPSOHV FROOHFWHG 7LVVXH VDPSOHV LQFOXGHG OLYHU OHJ VHPLPHPEUDQRVXVf DQG QHFN UKRPERLGHXVf PXVFOH EDFN IDW WK ULE DUHDf DQG OHDI IDW 7LVVXH VDPSOHV ZHUH IUR]HQ IROORZLQJ FROOHFWLRQ DQG VWRUHG DW r& XQWLO DQDO\]HG 9LWDPLQ ( DWRFRSKHUROf ZDV H[WUDFWHG IURP VHUXP VDPSOHV XVLQJ WKH SURFHGXUH GHVFULEHG E\ 0F0XUUD\ DQG %ODQFKIORZHU Df ZLWK PRGLILFDWLRQV GHVFULEHG E\ 1MHUX HW DO f ([WUDFWLRQ RI YLWDPLQ ( IURP WLVVXH DQG IHHG VDPSOHV ZDV GRQH XVLQJ WKH SURFHGXUH RI &KXQJ HW DO f 7KLV SURFHGXUH ZDV VLPLODU WR WKDW RI 0F0XUUD\ DQG %ODQFKIORZHU Ef DQG +DWDP DQG .D\GHQ f ZLWK

PAGE 40

PRGLILFDWLRQV 1MHUX HW DO f $OSKDWRFRSKHURO ZDV GHWHUPLQHG E\ LQMHFWLQJ XO RI WKH UHFRQVWLWXWHG VDPSOH VHUXP RU WLVVXHf LQWR DQ +3/& $QGHUVRQ HW DO ,Q 3UHVVf $OSKDWRFRSKHURO FRQFHQWUDWLRQ RI VDPSOHV ZDV FDOFXODWHG IURP WKH NQRZQ FRQFHQWUDWLRQ RI VWDQGDUGV 6SLNHG VDPSOHV RI DWRFRSKHURO ZHUH IRXQG WR KDYH D PHDQ UHFRYHU\ UDWH RI s b 9LWDPLQ $ ZDV H[WUDFWHG IURP VHUXP DQG WLVVXHV DV GHVFULEHG E\ &KHZ HW DO f DQG 0RRQH\ f ([WUDFWLRQ SURFHGXUHV ZHUH SHUIRUPHG XQGHU GDUN FRQGLWLRQV ZLWK HLWKHU \HOORZ ILOWHUHG RU VXEGXHG OLJKW 9LWDPLQ $ ZDV DVVD\HG DQG GHWHUPLQHG E\ WKH PHWKRG RI 0RRQH\ f 7KH RQO\ PRGLILFDWLRQV ZHUH WKDW +3/& SUHSDUHG VDPSOHV ZHUH HOXWHG XVLQJ DQ YROYROf PL[WXUH RI LVRRFWDQH WHWUDK\GURIXUDQ ZLWK b DFHWLF DFLG DQG UHWHQWLRQ WLPH ZDV DSSUR[LPDWHO\ PLQ $OO WUDQV UHWLQRO 6LJPD &KHPLFDO &R 6W /RXLV 02f VWDQGDUGV ZHUH SUHSDUHG DQG XVHG WR GHWHUPLQH FRQFHQWUDWLRQ RI VDPSOHV 6HYHUDO OLYHU VDPSOHV ZHUH VSLNHG WR GHWHUPLQH WKH UHFRYHU\ UDWH DQG YDOLGDWH WKH H[WUDFWLRQ SURFHGXUH 5HFRYHU\ UDWH RI UHWLQRO ZDV IRXQG WR EH s b 'DWD FROOHFWLRQ LQFOXGHG VHUXP DQG WLVVXH FRQFHQWUDWLRQV RI YLWDPLQ $ DQG ( DQG SHUIRUPDQFH GDWD ERG\ ZHLJKW JDLQ IHHGWRJDLQ DQG IHHG LQWDNHf 7LVVXH GDWD ZHUH ORJ WUDQVIRUPHG SULRU WR DQDO\VLV WR LPSURYH KRPRJHQHLW\ RI YDULDQFH $ XQLYDULDWH UHSHDWHG PHDVXUHV

PAGE 41

$129$ ZDV SHUIRUPHG RQ VHUXP GDWD 'DWD ZHUH DQDO\]HG XVLQJ WKH */0 SURFHGXUH RI 6$6 f 2UWKRJRQDO SRO\QRPLDO FRQWUDVWV ZHUH SHUIRUPHG WR FRPSDUH WUHDWPHQW PHDQV 5HVXOWV DQG 'LVFXVVLRQ *URZWK SHUIRUPDQFH GDWD RI WKH SLJV LQ WKLV VWXG\ DUH VXPPDUL]HG LQ 7DEOH 3LJV JUHZ ZHOO RQ DOO GLHWDU\ WUHDWPHQWV 15& f $Q LQFUHDVH LQ DYHUDJH GDLO\ JDLQ RI SLJV DSSURDFKHG VLJQLILFDQFH 3 f LQ OLQHDU IDVKLRQ DV GLHWDU\ YLWDPLQ ( LQFUHDVHG IHHG WR JDLQ ZDV QRW DIIHFWHG 3 f 'LHWDU\ YLWDPLQ $ OHYHOV KDG QR HIIHFW RQ SLJ SHUIRUPDQFH 3 f +RSSH HW DO f IHG SLJV ,8 RI VXSSOHPHQWDO YLWDPLQ ( LQ FRPELQDWLRQ ZLWK RU ,8 RI UHWLQRONJ RI GLHW DQG DOVR REVHUYHG QR GLIIHUHQFHV LQ SLJ SHUIRUPDQFH 2WKHU UHVHDUFKHUV KDYH QRWHG VLPLODU UHVXOWV LQ WKH FKLFN 6NODQ DQG 'RQRJKXH $EDZL DQG 6XOOLYDQ f DQG UDW %ODNHO\ HW DO f 6HUXP DWRFRSKHURO FRQFHQWUDWLRQV DUH UHSRUWHG LQ 7DEOHV DQG 6HUXP WRFRSKHURO FRQFHQWUDWLRQV ZHUH DIIHFWHG E\ GLHWDU\ DWRFRSKHURO RQ DOO VDPSOLQJ GD\V H[FHSW G 7DEOH f :KHQ QR VXSSOHPHQWDO YLWDPLQ ( ZDV DGGHG WR WKH GLHW WKHUH ZDV D VWHDG\ GHFOLQH LQ VHUXP WRFRSKHURO FRQFHQWUDWLRQ IURP G WR G 7DEOH f :KHQ WKH GLHW ZDV VXSSOHPHQWHG ZLWK ,8 RI DWRFRSKHU\O DFHWDWH SHU NJ RI GLHW VHUXP WRFRSKHURO FRQFHQWUDWLRQV GHFOLQHG 3 f

PAGE 42

7DEOH 3HUIRUPDQFH RI JURZLQJILQLVKLQJ VZLQH IHG GLHWV ZLWK GLIIHUHQW GLHWDU\ OHYHOV RI YLWDPLQ ( DQG 9LWDPLQ $ $GGHG 9LW ( ,8NJ $GGHG YLWDPLQ $ ,8NT 0HDQ 6( $ 7 7 ? }} QY\ XDLL\ \DLQ MV\ 0HDQ 6( QY\ XDLL\ 0HDQ 6( QY\ LFFX\DLLL 0HDQ 6( 1RWH 6HYHQ SHQV SHU WUHDWPHQW ZLWK SLJV SHU SHQ D3!) ( $ (r$ ( OLQHDU E3!) ( $ (r$ ( OLQHDU &3!)( $ (r$ ( OLQHDU

PAGE 43

7DEOH 0HDQ DWRFRSKHURO FRQFHQWUDWLRQV LQ EORRG VHUXP GXH WR GLHWDU\ DGGLWLRQV RI YLWDPLQV ( DQG $ 9LW ( DQG $ ,8NTD 'D\V E E 6( /WJPO 1RWH (DFK PHDQ LV EDVHG RQ REVHUYDWLRQV f9LWDPLQ ( OLQHDU HIIHFW 3 DOO GD\V H[FHSW G ( [ $ 3 G DQG RQO\ E ,8 RI UHWLQ\O DFHWDWH

PAGE 44

7DEOH 0DLQ PHDQV RI VHUXP DWRFRSKHURO GXH WR GLHWDU\ DGGLWLRQV RI YLWDPLQV ( DQG $ 'D\V 9LW (f ,8ND 9LW $E ,8ND 6( 6( IM/&M LQL 1RWH (DFK PHDQ LV EDVHG RQ RU REVHUYDWLRQV f9LWDPLQ ( OLQHDU HIIHFW 3 DOO GD\V H[FHSW G ( TXDGUDWLF 3 G DQG RQO\ E9LWDPLQ $ HIIHFW 3 G RQO\

PAGE 45

IURP WKHLU LQLWLDO FRQFHQWUDWLRQ E\ G DQG WKHQ VWDELOL]HG WKURXJKRXW WKH UHPDLQGHU RI WKH WULDO 7DEOH f 6HUXP WRFRSKHURO FRQFHQWUDWLRQ LQFUHDVHG 3 f ZLWK WKH KLJKHVW YLWDPLQ ( VXSSOHPHQWDWLRQ OHYHO E\ G DQG FRQWLQXHG WR LQFUHDVH 3 f WR G DIWHU ZKLFK VHUXP FRQFHQWUDWLRQ ZDV PDLQWDLQHG WKURXJKRXW WKH VWXG\ 7DEOH f 6HUXP WRFRSKHURO ZDV KLJKHVW 3 f DW WKH KLJKHVW OHYHO RI YLWDPLQ ( VXSSOHPHQWDWLRQ RQ DOO GD\V H[FHSW G 2YHUDOO DV GLHWDU\ OHYHOV RI VXSSOHPHQWDO YLWDPLQ ( LQFUHDVHG VHUXP FRQFHQWUDWLRQ RI WRFRSKHURO DOVR LQFUHDVHG OLQHDU 3 f 2WKHU VWXGLHV KDYH VKRZQ GLHWDU\ YLWDPLQ ( FRPSRXQGV DUH HIIHFWLYH LQ LQFUHDVLQJ EORRG WRFRSKHURO DQG WKDW EORRG FRQFHQWUDWLRQV LQFUHDVHG ZLWK LQFUHDVLQJ GLHWDU\ YLWDPLQ ( -HQVHQ HW DO $VJKDU HW DO DQG $QGHUVRQ HW DO ,Q 3UHVVf 6HUXP WRFRSKHURO FRQFHQWUDWLRQV GXH WR VXSSOHPHQWLQJ WKH GLHW ZLWK YLWDPLQ $ DW RU ,8NJ RI GLHW DUH VXPPDUL]HG LQ 7DEOH 3LJV IHG WKH ORZ OHYHO RI YLWDPLQ $ WHQGHG WR KDYH KLJKHU FRQFHQWUDWLRQV RI VHUXP WRFRSKHURO WKDQ SLJV IHG WKH KLJK OHYHO RI YLWDPLQ $ RQ DOO EORRG VDPSOLQJ GD\V EXW ZDV VLJQLILFDQW RQO\ RQ G 3 f $PRQJ WKH WKUHH GLHWDU\ OHYHOV RI YLWDPLQ ( WKH KLJKHVW GLHWDU\ YLWDPLQ ( WHQGHG WR EH GHFUHDVHG E\ WKH KLJK GLHWDU\ YLWDPLQ $ WR WKH JUHDWHVW H[WHQW EXW ZDV VLJQLILFDQWO\ ORZHU 3 f RQ G DQG G RQO\ 7DEOH f $OWKRXJK VLJQLILFDQW GLIIHUHQFHV ZHUH QRWHG DERYH WKH PDJQLWXGH RI

PAGE 46

WKHVH FKDQJHV ZDV YHU\ VPDOO 7KHUHIRUH YLWDPLQ $ OHYHO RI ,8NJ KDG RQO\ D PLQLPDO HIIHFW RQ EORRG WRFRSKHURO LQ VZLQH :HDYHU HW DO f LQ D WULDO ZLWK \RXQJ VWDUWLQJ SLJV REVHUYHG WKDW KLJK GLHWDU\ OHYHOV RI YLWDPLQ $ DQG ,8NJf IHG ZLWK ,8 RI DGGHG YLWDPLQ (NJ WHQGHG WR ORZHU SODVPD WRFRSKHURO FRQFHQWUDWLRQV VOLJKWO\ ,Q WULDOV ZLWK FKLFNV DQG UDWV KRZHYHU D GHFUHDVH LQ EORRG WRFRSKHURO ZDV QRWHG XSRQ IHHGLQJ GLHWV ZLWK H[FHVVLYH YLWDPLQ $ ,Q WKHVH WULDOV YHU\ KLJK YLWDPLQ $ FRQFHQWUDWLRQV ,8NJ RI GLHWf ZHUH IHG 6NODQ DQG 'RQRJKXH %ODNHO\ HW DO f 7KLV QHJDWLYH HIIHFW LV WKRXJKW WR EH DWWULEXWHG WR FRPSHWLWLRQ IRU DEVRUSWLRQ VLWHV LQ WKH VPDOO LQWHVWLQH RU HQKDQFHG R[LGDWLRQ RI WRFRSKHURO SULRU WR WRFRSKHURO UHDFKLQJ WKH VPDOO LQWHVWLQH 6NODQ DQG 'RQRJKXH f $OWKRXJK VRPH GLIIHUHQFHV ZHUH REVHUYHG RQ LQGLYLGXDO VDPSOLQJ GD\V VXSSOHPHQWDO YLWDPLQ $ KDG QR HIIHFW 3 f RQ VHUXP UHWLQRO 7DEOH f 'LIIHUHQFHV WKDW ZHUH QRWHG ZHUH YHU\ VPDOO DQG ZLWKLQ QRUPDO YDOXHV IRU VHUXP UHWLQRO XVXDOO\ IRXQG LQ WKH SLJ .DQHNR f %ORRG UHWLQRO ZDV DOVR QRW DIIHFWHG E\ GLHWDU\ VXSSOHPHQWDWLRQ RI YDULRXV OHYHOV RI YLWDPLQ $ LQ VWXGLHV GRQH E\ RWKHU UHVHDUFKHUV $EDZL DQG 6XOOLYDQ %ODNHO\ HW DO +RSSH HW DO f :LWK WZR H[FHSWLRQV WKHUH ZDV QR HIIHFW 3 f RQ VHUXP UHWLQRO GXH WR VXSSOHPHQWDWLRQ RI YLWDPLQ ( DW DQ\ RI

PAGE 47

7DEOH 0DLQ PHDQV RI VHUXP UHWLQRO GXH WR GLHWDU\ DGGLWLRQV RI YLWDPLQV ( DQG $ 9LW (D ,8NFU 'D\V 9LW $E LXND 6( 6( 0JPO &0 2 f 1RWH (DFK PHDQ LV EDVHG RQ RU REVHUYDWLRQV f9LWDPLQ ( OLQHDU HIIHFW 3 RQ G DQG RQO\ E9LWDPLQ $ HIIHFW 3 RQ G DQG RQO\

PAGE 48

WKH GLHWDU\ OHYHOV HYDOXDWHG LQ WKLV VWXG\ ,QFUHDVLQJ GLHWDU\ YLWDPLQ ( 7DEOH f UHVXOWHG LQ KLJKHU 3 f UHWLQRO VHUXP FRQFHQWUDWLRQ RQ G LQ SLJV IHG WKH KLJKHVW VXSSOHPHQWDO YLWDPLQ ( EXW DOVR UHVXOWHG LQ WKH ORZHVW 3 f VHUXP UHWLQRO FRQFHQWUDWLRQ RQ G $OWKRXJK ERWK ZHUH VLJQLILFDQW WKH GLIIHUHQFHV ZHUH VPDOO :HDYHU HW DO f LQ WULDOV ZLWK JURZLQJ SLJV DOVR QRWHG WKDW SODVPD YLWDPLQ $ ZDV QRW DIIHFWHG E\ GLHWDU\ OHYHO RI YLWDPLQ ( 7LVVXH DWRFRSKHURO LQFUHDVHG OLQHDU 3 f DV GLHWDU\ YLWDPLQ ( LQFUHDVHG 7DEOH f $V GLHWDU\ YLWDPLQ ( LQFUHDVHG IURP WR ,8 SHU NJ RI GLHW WLVVXH WRFRSKHURO LQFUHDVHG E\ DW OHDVW D IDFWRU RI RU PRUH LQ DOO WLVVXHV HYDOXDWHG 7KLV ILQGLQJ LV LQ DJUHHPHQW ZLWK RWKHUV ZKR KDYH REVHUYHG VLPLODU UHVSRQVHV LQ SLJV -HQVHQ HW DO $VJKDU HW DO f $PRQJ WKH WLVVXHV VDPSOHG WKH KLJKHVW FRQFHQWUDWLRQ RI DWRFRSKHURO GXH WR WUHDWPHQW ZDV IRXQG LQ WKH DGLSRVH WLVVXH IROORZHG E\ OLYHU DQG PXVFOH WLVVXH UHVSHFWLYHO\ 7DEOH f +LJK VXSSOHPHQWDWLRQ RI YLWDPLQ $ KDG QR HIIHFW 3 f RQ WRFRSKHURO FRQFHQWUDWLRQ LQ DQ\ RI WKH WLVVXHV VWXGLHG 7DEOH f 7KH UHWLQRO FRQFHQWUDWLRQ LQ WKH OLYHU ZDV JUHDWO\ HQKDQFHG 3 f ZKHQ SLJV ZHUH IHG WKH KLJK OHYHO RI YLWDPLQ $ 7DEOH f 5HWLQRO FRQFHQWUDWLRQ LQ RWKHU WLVVXHV EDFN IDW OHDI IDW UKRPERLGHXV DQG VHPLPHPEUDQRVXVf ZDV QRW GHWHFWHG DQG DUH QRW UHSRUWHG 'LHWDU\ YLWDPLQ ( KDG QR HIIHFW 3 f RQ OLYHU UHWLQRO

PAGE 49

7DEOH 0DLQ PHDQV RI WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV GXH WR GLHWDU\ DGGLWLRQV RI YLWDPLQV ( DQG $ 7LVVXH 9LW (D ,8ND 6( 9LW $ ,8ND 6( 0JJ 6HPLPHPEUDQRVXV D E & 5KRPERLGHXV D E & %DFN IDW ,D E & /HDI IDW D OE R R R U+ /LYHU D D E 1RWH (DFK PHDQ LV EDVHG RQ RU REVHUYDWLRQV D9LWDPLQ ( OLQHDU HIIHFW 3 IRU DOO WLVVXHV 2 R

PAGE 50

7DEOH 0DLQ PHDQV RI WLVVXH UHWLQRO FRQFHQWUDWLRQV GXH WR GLHWDU\ DGGLWLRQV RI YLWDPLQV ( DQG $ 7LVVXH 9LW ( ,8NFU 9LW $D ,8ND 6( 6( /LYHU 0JJ 1RWH (DFK PHDQ LV EDVHG RQ RU REVHUYDWLRQV f9LWDPLQ $ HIIHFW 3

PAGE 51

FRQFHQWUDWLRQ 7DEOH f +RSSH HW DO f IRXQG WKDW OLYHU UHWLQRO ZDV OLQHDUO\ UHODWHG WR GLHWDU\ UHWLQRO LQ D WULDO LQYROYLQJ \RXQJ JURZLQJ SLJV DV ZDV DOVR REVHUYHG LQ WKH SUHVHQW VWXG\ 7KH\ DOVR IRXQG WKDW ,8 RI GLHWDU\ UHWLQRO GLG QRW DIIHFW WLVVXH DWRFRSKHURO RWKHU WKDQ WKH KHDUW LQ ZKLFK DWRFRSKHURO ZDV VOLJKWO\ GHSUHVVHG +RZHYHU 3XGHONLHZLF] HW DO f IHHGLQJ YLWDPLQ $ GHSOHWHG FKLFNV DQG ,8 RI YLWDPLQ $ DFHWDWHNJ RI GLHW REVHUYHG D PDUNHG GHFOLQH LQ WRFRSKHURO FRQFHQWUDWLRQ LQ OLYHU WLVVXH DW WKH KLJKHVW GLHWDU\ YLWDPLQ $ OHYHOV ,Q FRQFOXVLRQ WKHUH ZDV QR FRQVLVWHQW HYLGHQFH WKDW H[FHVVLYH GLHWDU\ YLWDPLQ $ ,8NJ RI GLHWf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

PAGE 52

YLWDPLQ $NJ RI GLHW 7KH WULDO LQYROYHG FURVVEUHG SLJV NJf GLYLGHG E\ VH[ ZHLJKW DQG JHQHWLF EDFNJURXQG LQWR SHQV RI WZR SLJV HDFK 7UHDWPHQW ZDV DVVLJQHG DW UDQGRP WR SHQV ZLWKLQ HDFK RI VHYHQ UHSOLFDWLRQV 3LJV ZHUH IHG JURZHU GLHWV b O\VLQHf XQWLO WKH\ UHDFKHG NJ DYHUDJH ZHLJKW DQG ZHUH WKHQ VZLWFKHG WR ILQLVKHU GLHWV bf XQWLO NJ 6HUXP ZDV FROOHFWHG RQ GD\ DQG RI WKH IHHGLQJ SHULRG 7LVVXH VDPSOHV OLYHU PXVFOH EDFN IDW DQG OHDI IDWf ZHUH FROOHFWHG IURP RQH SLJ EDUURZf LQ HDFK SHQ DW WKH HQG RI WKH IHHGLQJ SKDVH 2YHUDOO DYHUDJH GDLO\ JDLQ DQG IHHGWRJDLQ ZHUH NJ DQG UHVSHFWLYHO\ ZLWKRXW WUHDWPHQW GLIIHUHQFHV 3 f ([FHVVLYH GLHWDU\ YLWDPLQ $ KDG QR HIIHFW 3 f RQ VHUXP UHWLQRO FRQFHQWUDWLRQV H[FHSW G LQ ZKLFK WKHUH ZDV D VPDOO 3 f LQFUHDVH 6HUXP WRFRSKHURO ZDV LQFUHDVHG 3 OLQHDUf E\ G ZLWK GLHWDU\ YLWDPLQ ( VXSSOHPHQWDWLRQ DQG ZDV PDLQWDLQHG 3 f WKURXJKRXW WKH IHHGLQJ SHULRG +LJK GLHWDU\ YLWDPLQ $ UHVXOWHG LQ D VPDOO EXW VLJQLILFDQW 3 f GHFUHDVH LQ VHUXP WRFRSKHURO RQ G VHUXP WRFRSKHURO FRQFHQWUDWLRQV ZHUH QRW DIIHFWHG RQ RWKHU GD\V 7LVVXH WRFRSKHURO ZDV LQFUHDVHG 3 OLQHDUf DV GLHWDU\ YLWDPLQ ( LQFUHDVHG IURP WR ,8NJ /LYHU UHWLQRO LQFUHDVHG 3 f E\ D IDFWRU RI HLJKW 1R FRQVLVWHQW HYLGHQFH ZDV IRXQG WKDW KLJK GLHWDU\ YLWDPLQ $ LQWHUIHUHG ZLWK SHUIRUPDQFH RU ZLWK VHUXP RU WLVVXH WRFRSKHURO LQ JURZLQJILQLVKLQJ VZLQH

PAGE 53

&+$37(5 ())(&7 2) ,1-(&7(' 9,7$0,1 $ $1' ',(7$5< 6833/(0(17$7,21 2) 9,7$0,1 ( 21 5(352'8&7,9( 3(5)250$1&( $1' 72&23+(52/ 67$786 ,1 *(67$7,1* *,/76 ,QWURGXFWLRQ 6XSSOHPHQWDO YLWDPLQ $ DQGRU FDURWHQH JLYHQ YLD LQMHFWLRQ MXVW EHIRUH DQGRU VKRUWO\ DIWHU EUHHGLQJ DSSHDUV WR HQKDQFH UHSURGXFWLYH SHUIRUPDQFH RI JLOWV DQG VRZV %ULHI DQG &KHZ &RIIH\ DQG %ULWW f 7KHUH LV HYLGHQFH WKDW KLJK GLHWDU\ YLWDPLQ $ PD\ LQWHUIHUH ZLWK ERWK YLWDPLQ ( DEVRUSWLRQ DQG EORRG DWRFRSKHURO FRQFHQWUDWLRQV +LJK GLHWDU\ YLWDPLQ $ UHGXFHG DEVRUSWLRQ RI DWRFRSKHURO LQ WULDOV ZLWK FKLFNV 6NODQ DQG 'RQRJKXH f $EDZL DQG 6XOOLYDQ f QRWHG D GHFUHDVH LQ SODVPD YLWDPLQ ( FRQFHQWUDWLRQV ZKHQ GHSOHWHG FKLFNV ZHUH DGPLQLVWHUHG KLJK ,8NJf OHYHOV RI GLHWDU\ YLWDPLQ $ %ODNHO\ HW DO f DOVR UHSRUWHG WKDW KLJK OHYHOV RI YLWDPLQ $ WLPHV UHJXLUHPHQWf SOXV KLJK OHYHOV RI %FDURWHQH PJNJ RI GLHWf UHGXFHG YLWDPLQ ( SODVPD FRQFHQWUDWLRQ E\ b LQ UDWV 7KLV VWXG\ ZDV FRQGXFWHG WR HYDOXDWH WKH HIIHFW RI LQMHFWLQJ YLWDPLQ $ MXVW EHIRUH GXULQJ DQG VKRUWO\ DIWHU EUHHGLQJ DQG GLHWDU\ VXSSOHPHQWDWLRQ RI YLWDPLQ ( RQ UHSURGXFWLYH SHUIRUPDQFH DQG RQ EORRG DQG WLVVXH

PAGE 54

FRQFHQWUDWLRQV RI DWRFRSKHURO GXULQJ HDUO\ JHVWDWLRQ RI JLOWV ([SHULPHQWDO 3URFHGXUHV 7KH WULDO ZDV D [ IDFWRULDO GHVLJQ DQG LQYROYHG WR PRQWK ROGf FURVVEUHG JLOWV 7KH JLOWV ZHUH GLYLGHG LQWR SHQV RI JLOWV HDFK *LOWV XVHG LQ WKLV VWXG\ ZHUH IURP D SUHYLRXV WULDO WKDW LQYROYHG WKH IHHGLQJ RI GLHWV VXSSOHPHQWHG ZLWK HLWKHU /f RU +f ,8 YLWDPLQ $NJ RI GLHW 7KLV ZDV WDNHQ LQWR FRQVLGHUDWLRQ LQ WKH DOORWPHQW RI JLOWV WR WUHDWPHQW / DQG + SHU SHQf (DFK SHQ ZDV UDQGRPO\ DVVLJQHG WR RQH RI IRXU WUHDWPHQWV 7UHDWPHQWV FRQVLVWHG RI D EDVDO FRUQ VR\EHDQ PHDO GLHW 7DEOH f VXSSOHPHQWHG ZLWK '/DWRFRSKHU\ DFHWDWH +RIIPDQQ/D 5RFKH ,QF 1XWOH\ 1-f DW OHYHOV RI HLWKHU RU ,8NJ RI GLHW *LOWV ZHUH IHG H[SHULPHQWDO GLHWV EHJLQQLQJ G SULRU WR EUHHGLQJ WKURXJK G RI JHVWDWLRQ +DOI RI WKH JLOWV ZHUH JLYHQ WKUHH LQMHFWLRQV LP LQ WKH QHFNf RI ,8 HDFK RI YLWDPLQ $ YLWDPLQ $ SDOPLWDWH +RIIPDQQ/D 5RFKH ,QF 1XWOH\ 1-f WKH RWKHU KDOI ZHUH LQMHFWHG ZLWK YHKLFOH RQO\ 7KH JLOWV ZHUH LQMHFWHG DW G SUHEUHHGLQJ G f DW EUHHGLQJ G f DQG G SRVWEUHHGLQJ G f *LOWV ZHUH IHG NJ RI IHHGKG RQFH GDLO\ DQG JLYHQ IUHH DFFHVV WR ZDWHU *LOWV ZHUH KRXVHG LQ DQ RSHQn VLGHG EXLOGLQJ ZLWK VROLG FRQFUHWH IORRUV *LOWV ZHUH FKHFNHG WZLFH GDLO\ IRU HVWUXV GXULQJ WKH WULDO DQG GRXEOHG

PAGE 55

7DEOH &RPSRVLWLRQ RI GLHW IHG WR JHVWDWLQJ JLOWV ,QJUHGLHQW b *URXQG FRUQ 6R\EHDQ PHDO bf '\QDIRV *URXQG OLPHVWRQH 6DOW 7UDFH PLQHUDOf 9LWDPLQ SUHPL[E 6H SUHPL[F f3URYLGHG SSP ]LQF SSP FRSSHU SSP SSP LRGLQH LURQ DQG SSP PDQJDQHVH SSP FREDOW E3URYLGHG PJ ULERIODYLQ PJ QLDFLQ PJ SDQWRWKHQLF DFLG PJ FKROLQH FKORULGH XJ YLWDPLQ % PJ YLWDPLQ ,8 YLWDPLQ DQG ,8 YLWDPLQ $ SHU NJ RI GLHW F3URYLGHG SSP VHOHQLXP

PAGE 56

PDWHG RQ WKHLU VHFRQG RU WKLUG REVHUYHG HVWUXV WR GXURF [ KDPSVKLUH [ \RUNVKLUH ERDUV %ORRG VDPSOHV ZHUH FROOHFWHG E\ MXJXODU YHLQ SXQFWXUH IURP HDFK JLOW RQ G DQG RI JHVWDWLRQ WR PRQLWRU WKH VHUXP DWRFRSKHURO YLWDPLQ (f DQG UHWLQRO YLWDPLQ $f FRQFHQWUDWLRQV %ORRG VDPSOHV ZHUH FRYHUHG ZLWK IRLO WR SUHYHQW H[SRVXUH WR GLUHFW VXQOLJKW WDNHQ WR WKH ODERUDWRU\ FHQWULIXJHG DQG WKH VHUXP KDUYHVWHG 6HUXP ZDV VWRUHG DW r& XQWLO DQDO\]HG IRU DWRFRSKHURO DQG UHWLQRO FRQFHQWUDWLRQ 'XULQJ VWRUDJH VHUXP VDPSOHV ZHUH FRYHUHG ZLWK IRLO WR SUHYHQW H[SRVXUH WR OLJKW *LOWV ZHUH VODXJKWHUHG IROORZLQJ DFFHSWHG VODXJKWHU SURFHGXUHV RQ G RI JHVWDWLRQ DW WKH 8QLYHUVLW\ RI )ORULGD PHDWV ODERUDWRU\ 5HSURGXFWLYH WUDFWV ZHUH LPPHGLDWHO\ UHPRYHG DQG UHIULJHUDWHG IRU ODWHU FRXQWLQJ RI FRUSRUD OWHD &/f DQG UHFRYHU\ RI HPEU\RV 7LVVXH VDPSOHV ZHUH DOVR FROOHFWHG ZKLFK FRQVLVWHG RI HQGRPHWULXP HPEU\R RYDU\ XWHUXV OLYHU OHDI IDW EDFN IDW DQG PXVFOH VHPLPHPEUDQRVXV DQG UKRPERLGHXVf 7LVVXH VDPSOHV ZHUH VWRUHG DW r& XQWLO DQDO\]HG IRU DWRFRSKHURO DQG UHWLQRO FRQFHQWUDWLRQV 7KH WULDO ZDV FDUULHG RXW GXULQJ WKH VXPPHU DQG HDUO\ IDOO -XO\ WKURXJK 2FWREHUf 3LJV ZHUH PDQDJHG IROORZLQJ DFFHSWDEOH FDUH DQG PDQDJHPHQW SUDFWLFHV WKURXJKRXW WKH VWXG\ 3URWRFRO IRU DQLPDO FDUH KDG EHHQ DSSURYHG E\ WKH 8QLYHUVLW\ $QLPDO 8VH &RPPLWWHH 7KUHH SLJV IURP GLIIHUHQW

PAGE 57

WUHDWPHQW JURXSV ZHUH HOLPLQDWHG IURP WKH VWXG\ GXH WR GHDWK ODPHQHVV DQG IDLOXUH WR FRQFHLYH $OSKDWRFRSKHURO ZDV H[WUDFWHG IURP VHUXP DQG WLVVXHV XVLQJ SURFHGXUHV DV GHVFULEHG HDUOLHU $QGHUVRQ HW DO ,Q 3UHVVf $OSKDWRFRSKHURO FRQFHQWUDWLRQ ZDV GHWHUPLQHG E\ LQMHFWLQJ XO RI WKH H[WUDFWHG VDPSOH VHUXP DQG WLVVXHf LQWR WKH +3/& V\VWHP 5HWLQRO ZDV H[WUDFWHG DVVD\HG DQG FRQFHQWUDWLRQ GHWHUPLQHG IURP WKH VHUXP DQG WLVVXHV E\ WKH PHWKRG DV GHVFULEHG SUHYLRXVO\ 0RRQH\ f ([SHULPHQWDO GDWD LQFOXGHG VHUXP DQG WLVVXH FRQFHQWUDWLRQV RI DWRFRSKHURO DQG UHWLQRO DQG UHSURGXFWLYH SHUIRUPDQFH 7LVVXH GDWD ZDV ORJ WUDQVIRUPHG SULRU WR DQDO\VLV WR LPSURYH KRPRJHQHLW\ RI YDULDQFH $ XQLYDULDWH UHSHDWHG PHDVXUHV $129$ ZDV SHUIRUPHG RQ VHUXP GDWD 6$6 f 'DWD ZHUH DQDO\]HG DV D ; IDFWRULDO GHVLJQ ZLWK WKH IDFWRUV EHLQJ GLHWDU\ YLWDPLQ ( OHYHO DQG ZKHWKHU RU QRW JLOWV ZHUH LQMHFWHG ZLWK YLWDPLQ $ 5HVXOWV DQG 'LVFXVVLRQ /HYHOV RI YLWDPLQ ( XVHG ZHUH FKRVHQ WR UHIOHFW 15& f UHTXLUHPHQW DQG WR JLYH D YHU\ KLJK OHYHO 7KH LQMHFWHG OHYHO IRU YLWDPLQ $ ZDV FKRVHQ EHFDXVH LW ZDV WKRXJKW WR EH WKH XSSHU OLPLW WKDW ZRXOG HOLFLW D UHVSRQVH DQG QRW EH WR[LF WR WKH JLOWV DOVR VRPH HYLGHQFH RI WKLV GRVDJH ZDV EHLQJ XVHGf 5HSURGXFWLYH SHUIRUPDQFH GDWD DUH VXPPDUL]HG LQ 7DEOH

PAGE 58

7DEOH 0HDQ UHSURGXFWLYH UHVSRQVH FULWHULD RI JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ ,WHP 9LW ( ,8NJ DQG 9LW $ LQL f 6( 1R
PAGE 59

$OWKRXJK QRQH RI WKH GLIIHUHQFHV QRWHG ZLWK UHSURGXFWLYH GDWD ZHUH VLJQLILFDQW 3 f GXH WR WKH VPDOO QXPEHU RI JLOWV SHU WUHDWPHQW DQG WKH LQKHUHQW QDWXUH RI VZLQH UHSURGXFWLYH GDWD QHYHUWKHOHVV VRPH SRVLWLYH WUHQGV ZHUH REVHUYHG GXH WR WUHDWPHQW *LOWV UHFHLYLQJ WKH KLJK YLWDPLQ ( DQG KLJK YLWDPLQ $ WUHDWPHQW KDG ODUJHU 3 f OLWWHUV WKDQ JLOWV JLYHQ RWKHU WUHDWPHQWV (PEU\RQLF VXUYLYDO ZDV b LQ JLOWV JLYHQ WKH ORZ GLHWDU\ YLWDPLQ ( ZLWK QR LQMHFWHG YLWDPLQ $ DQG b LQ JLOWV JLYHQ WKH KLJK GLHWDU\ YLWDPLQ ( ZLWK QR LQMHFWHG $ (PEU\RQLF VXUYLYDO ZDV b LQ JLOWV LQMHFWHG ZLWK YLWDPLQ $ DQG IHG HLWKHU WKH ORZ RU KLJK YLWDPLQ ( GLHWV %ULHI DQG &KHZ f UHSRUWHG ODUJHU OLWWHU VL]H DQG KLJKHU HPEU\RQLF VXUYLYDO LQ JLOWV UHFHLYLQJ ZHHNO\ LQMHFWLRQV RI YLWDPLQ $ ,8f DQG % FDURWHQH PJf FRPSDUHG WR JLOWV IHG YLWDPLQ $ DQG IRn FDU RWHQH DW WKH VDPH OHYHOV +RZHYHU WKHVH JLOWV LQ WKH VWXG\ RI %ULHI DQG &KHZ f ZHUH GHSOHWHG RI YLWDPLQ $ DQG %FDURWHQH IRU ZHHNV EHIRUH WKH VWDUW RI WKH VWXG\ &RIIH\ DQG %ULWW f REVHUYHG RQ DYHUDJH SLJ LQFUHDVH LQ WKH QXPEHU RI SLJV ERUQ DOLYH DQG KLJKHU HPEU\RQLF VXUYLYDO LQ VRZV JLYHQ LP LQMHFWLRQV RI YLWDPLQ $ SDOPLWDWH ,8f FRPSDUHG WR VRZV JLYHQ YHKLFOH RQO\ FRUQ RLOf RQ GD\ RI ZHDQLQJ PDWLQJ DQG G SRVWEUHHGLQJ 7KHVH VRZV ZHUH DOVR VXSSOHPHQWHG ZLWK ,8 RI YLWDPLQ $ DFHWDWH SHU NJ RI GLHW 6HUXP FRQFHQWUDWLRQV RI DWRFRSKHURO LQ JLOWV IHG GLHWV

PAGE 60

VXSSOHPHQWHG ZLWK WZR OHYHOV RI YLWDPLQ ( ZLWK DQG ZLWKRXW LQMHFWHG YLWDPLQ $ DUH UHSRUWHG LQ 7DEOHV DQG ,QLWLDO DWRFRSKHURO VHUXP FRQFHQWUDWLRQV LQ JLOWV ZHUH VLPLODU G f :KHQ GLHWDU\ OHYHOV ZHUH LQFUHDVHG IURP WR ,8 RI '/DWRFRSKHU\O DFHWDWH SHU NJ RI GLHW VHUXP DWRFRSKHURO FRQFHQWUDWLRQV LQFUHDVHG 3 f E\ G DQG ZHUH PDLQWDLQHG WKURXJKRXW WKH GXUDWLRQ RI WKH VWXG\ 7DEOH f 7KLV ILQGLQJ LV LQ DJUHHPHQW ZLWK RWKHU VWXGLHV WKDW KDYH VKRZQ WKDW GLHWDU\ YLWDPLQ ( FRPSRXQGV DUH HIIHFWLYH LQ LQFUHDVLQJ VHUXP WRFRSKHURO DQG WKDW VHUXP FRQFHQWUDWLRQ LQFUHDVHG ZLWK LQFUHDVLQJ GLHWDU\ YLWDPLQ ( -HQVHQ HW DO $VJKDU HW DO 0DKDQ DQG $QGHUVRQ HW DO ,Q 3UHVVf 6HUXP DWRFRSKHURO ZDV QRW DIIHFWHG 3 f E\ LQMHFWLQJ JLOWV ZLWK YLWDPLQ $ UHWLQ\O SDOPLWDWHf H[FHSW RQ G 7DEOH f 2Q G WKH JLOWV IHG WKH ORZ YLWDPLQ ( GLHW KDG VLPLODU 3 f VHUXP WRFRSKHURO FRQFHQWUDWLRQV UHJDUGOHVV RI YLWDPLQ $ WUHDWPHQW ZKHUHDV JLOWV IHG WKH KLJK YLWDPLQ ( GLHW KDG KLJKHU 3 f VHUXP WRFRSKHURO FRQFHQWUDWLRQV ZKHQ LQMHFWHG ZLWK YLWDPLQ $ WKDQ JLOWV QRW LQMHFWHG ZLWK YLWDPLQ $ 7DEOH f 6HUXP FRQFHQWUDWLRQV RI UHWLQRO GXH WR WUHDWPHQW DUH VKRZQ LQ 7DEOHV DQG ,Q JHQHUDO WKHUH ZDV QR FRQVLVWHQW HIIHFW RQ VHUXP UHWLQRO GXH WR GLHWDU\ YLWDPLQ ( VXSSOHPHQWDWLRQ OHYHO RU LQMHFWLRQ RI YLWDPLQ $ +RZHYHU D GLIIHUHQFH LQ VHUXP UHWLQRO 3 f ZDV REVHUYHG RQ G LQ

PAGE 61

7DEOH 0HDQ VHUXP DWRFRSKHURO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 6DPSOLQJ GD\ 9LW ( ,8NFI DQG YLW $ LQL fEF 1R
PAGE 62

7DEOH 0DLQ PHDQ VHUXP DWRFRSKHURO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 6DPSOLQJ GD\ 9LW ( ,8NDf 9LW $ LQL EF 6( 1R
PAGE 63

7DEOH 0HDQ VHUXP UHWLQRO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 6DPSOLQJ GD\ 9LW ( ,8NFI DQG YLW $ LQL EF 6( 1R
PAGE 64

7DEOH 0DLQ PHDQ VHUXP UHWLQRO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 6DPSOLQJ GD\ 9LW ( ,8NDf 9LW $ LQL EF 6( 12
PAGE 65

WKDW VHUXP UHWLQRO FRQFHQWUDWLRQ ZDV KLJKHVW LQ JLOWV IHG WKH KLJK YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ DQG ORZHVW LQ JLOWV IHG ORZ YLWDPLQ ( ZLWKRXW LQMHFWHG $ 7KH GLIIHUHQFH QRWHG ZDV YHU\ VPDOO DQG ZLWKLQ QRUPDO YDOXHV IRU VHUXP UHWLQRO FRQFHQWUDWLRQV XVXDOO\ IRXQG LQ WKH SLJ 2XU ILQGLQJV DJUHH ZLWK :HDYHU HW DO f LQ WKDW SODVPD YLWDPLQ $ FRQFHQWUDWLRQ ZDV QRW DIIHFWHG E\ GLHWDU\ OHYHO RI YLWDPLQ ( 6HUXP UHWLQRO FRQFHQWUDWLRQV LQ WKH FKLFN DQG UDW KDYH DOVR EHHQ UHSRUWHG QRW WR EH DIIHFWHG E\ GLHWDU\ VXSSOHPHQWDWLRQ RI YLWDPLQ ( LQ VWXGLHV GRQH E\ $EDZL DQG 6XOOLYDQ f DQG %ODNHO\ HW DO f UHVSHFWLYHO\ 0RRQH\ f LQMHFWHG JLOWV ZLWK YLWDPLQ $ SDOPLWDWH UDQJLQJ IURP WR ,8 JLYHQ RQFH ZHHNO\f RU FDURWHQH WR PJf DQG REVHUYHG QR GLIIHUHQFH LQ SODVPD FRQFHQWUDWLRQV RI HLWKHU UHWLQRO RU FDURWHQH ,Q FRQWUDVW %ULHI DQG &KHZ f QRWHG LQFUHDVHG SODVPD YLWDPLQ $ FRQFHQWUDWLRQ XSRQ LQMHFWLQJ YLWDPLQ $ KRZHYHU JLOWV XVHG LQ WKHLU UHVHDUFK ZHUH GHSOHWHG RI YLWDPLQ $ SULRU WR WKH VWXG\ DQG WKH\ DOVR UHFHLYHG LQMHFWHG FDURWHQH 6HUXP UHWLQRO PD\ KDYH EHHQ HOHYDWHG ZLWK LQMHFWLRQ RI YLWDPLQ $ HDUO\ LQ RXU VWXG\ EXW PD\ KDYH EHHQ PLVVHG VLQFH EORRG VDPSOHV ZHUH WDNHQ G DIWHU LQMHFWLRQ 7LVVXH DWRFRSKHURO FRQFHQWUDWLRQ LQ JHVWDWLQJ JLOWV LQFUHDVHG 3 f DV GLHWDU\ VXSSOHPHQWDWLRQ RI YLWDPLQ ( LQFUHDVHG 7DEOH f LQ DOO WLVVXHV H[FHSW DGLSRVH 7KLV ILQGLQJ LV LQ DJUHHPHQW ZLWK RWKHUV ZKR KDYH REVHUYHG

PAGE 66

7DEOH 0HDQ WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV RI JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 7LVVXH 9LW ( ,8NJ DQG YLW $ LQME 1R
PAGE 67

VLPLODU UHVSRQVHV LQ WKH SLJ -HQVHQ HW DO $VJKDU HW DO 0DKDQ f $YHUDJH WRFRSKHURO FRQFHQWUDWLRQ LQFUHDVHG E\ D IDFWRU RI LQ HPEU\RV XSRQ KLJK GLHWDU\ VXSSOHPHQWDWLRQ LQGLFDWLQJ WKDW WRFRSKHURO LV WUDQVIHUUHG IURP WKH GDP WR WKH GHYHORSLQJ HPEU\R 9LWDPLQ $ LQMHFWLRQV KDG QR HIIHFW 3 f RQ WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV H[FHSW LQ WKH HQGRPHWULXP ZKHUH WKHUH ZDV D YLWDPLQ ( [ YLWDPLQ $ LQWHUDFWLRQ 3 f ,Q WKH HQGRPHWULXP JLOWV IHG ORZ YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ KDG VOLJKWO\ ORZHUHG WRFRSKHURO FRQFHQWUDWLRQ ZKLOH JLOWV IHG WKH KLJK YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ KDG LQFUHDVHG WRFRSKHURO FRQFHQWUDWLRQ RYHU WKDW RI WKH QRQ LQMHFWHG JLOWV 7DEOH f 9LWDPLQ $ LQMHFWLRQV DSSHDU WR KDYH QR HIIHFW RQ WKH WUDQVIHU RI WRFRSKHURO LQWR WKH GHYHORSLQJ HPEU\RV DV WRFRSKHURO FRQFHQWUDWLRQ LQ WKH HPEU\RV ZDV QRW LQIOXHQFHG E\ YLWDPLQ $ LQMHFWLRQ 3 f ,QMHFWLQJ YLWDPLQ $ KDG QR HIIHFW 3 f RQ UHWLQRO FRQFHQWUDWLRQ LQ DQ\ RI WKH WLVVXHV VWXGLHG LQFOXGLQJ WKH OLYHU 5HWLQRO FRQFHQWUDWLRQV LQ WLVVXHV RWKHU WKDQ OLYHU KRZHYHU ZHUH YHU\ VPDOO RU QRQH[LVWHQW $YHUDJH FRQFHQWUDWLRQ LQ WKH OLYHU ZDV AJJ 0RRQH\ f IRXQG QR GLIIHUHQFHV LQ FRQFHQWUDWLRQ RI UHWLQRO LQ XWHULQH IOXVKLQJV LQ JLOWV WKDW ZHUH LQMHFWHG ZLWK YDU\LQJ OHYHOV RI YLWDPLQ $ OLYHU UHWLQRO ZDV QRW GHWHUPLQHG $PRQJ WKH WLVVXHV VDPSOHG WKH KLJKHVW DYHUDJH FRQFHQWUDWLRQ RI DWRFRSKHURO XSRQ VXSSOHPHQWDWLRQ RI KLJK

PAGE 68

OHYHO RI YLWDPLQ ( ZDV IRXQG LQ WKH RYDU\ IROORZHG E\ OLYHU DGLSRVH UKRPERLGHXV DQG HQGRPHWULXP UHVSHFWLYHO\ 7DEOH f 7KUHH RWKHU WLVVXHV IROORZHG VHPLPHPEUDQRVXV RYLGXFW XWHUXVf WKDW KDG VLPLODU DYHUDJH FRQFHQWUDWLRQV DQG HPEU\R WLVVXH KDG WKH ORZHVW DWRFRSKHURO FRQFHQWUDWLRQ XQLWV SHU ZHW WLVVXH EDVLVf ,Q FRQFOXVLRQ WKHUH ZDV QR FRQVLVWHQW HYLGHQFH IRXQG LQ WKLV VWXG\ WKDW LQMHFWLQJ D UHODWLYHO\ ODUJH DPRXQW RI YLWDPLQ $ LQMHFWLRQV RI ,8f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

PAGE 69

7DEOH 0DLQ PHDQ WLVVXH DWRFRSKHURO FRQFHQWUDWLRQV LQ JHVWDWLQJ JLOWV JLYHQ GLHWDU\ DGGLWLRQV RI YLWDPLQ ( DQG LQMHFWHG ZLWK YLWDPLQ $ 7LVVXHV 9LW ( ,8NJ 9LW $ LQME 1R
PAGE 70

PHDO EDVHG GLHWV VXSSOHPHQWHG ZLWK '/DWRFRSKHU\ DFHWDWH WR SURYLGH HLWKHU RU ,8 RI DGGHG YLWDPLQ (NJ RI GLHW *LOWV ZHUH IHG GDLO\ NJK EHJLQQLQJ G SUHEUHHGLQJ WKURXJK G RI JHVWDWLRQ +DOI RI WKH JLOWV ZHUH LQMHFWHG LPf ZLWK ,8 RI UHWLQRO SDOPLWDWH DW G SUHEUHHGLQJ EUHHGLQJ G f DQG G SRVWEUHHGLQJ WKH RWKHU KDOI ZHUH LQMHFWHG ZLWK YHKLFOH RQO\ $OO JLOWV ZHUH GRXEOH PDWHG GXULQJ WKHLU VHFRQG RU WKLUG HVWUXV %ORRG VDPSOHV ZHUH FROOHFWHG RQ G f§ DQG RI JHVWDWLRQ *LOWV ZHUH VODXJKWHUHG RQ G RI JHVWDWLRQ IROORZLQJ DFFHSWHG VODXJKWHU SURFHGXUHV 7ZHQW\QLQH JLOWV FRQFHLYHG 7KH QXPEHU RI FRUSRUD OWHD DQG HPEU\RV ZDV QRW DIIHFWHG 3 f E\ WUHDWPHQW 6HUXP WRFRSKHURO FRQFHQWUDWLRQV LQFUHDVHG ZLWK ,8 RI YLWDPLQ ( E\ G DQG ZHUH VWDEOH WKURXJK G RI JHVWDWLRQ 3 f 9LWDPLQ $ LQMHFWLRQV KDG QR HIIHFW 3 f RQ VHUXP WRFRSKHURO FRQFHQWUDWLRQV H[FHSW RQ G ZKHQ D VPDOO LQFUHDVH 3 f ZDV QRWHG +LJK GLHWDU\ YLWDPLQ ( LQFUHDVHG WRFRSKHURO FRQFHQWUDWLRQ 3 f LQ DOO WLVVXHV H[DPLQHG H[FHSW IDW $ YLWDPLQ ( [ YLWDPLQ $ LQWHUDFWLRQ 3 f ZDV QRWHG LQ HQGRPHWULXP WLVVXH /RZ GLHWDU\ YLWDPLQ ( DQG LQMHFWLRQV RI YLWDPLQ $ VOLJKWO\ ORZHUHG WRFRSKHURO FRQFHQWUDWLRQ ZKLOH KLJK YLWDPLQ ( DQG YLWDPLQ $ LQMHFWLRQV LQFUHDVHG WRFRSKHURO FRQFHQWUDWLRQ LQ WKH HQGRPHWULXP ,QFUHDVLQJ GLHWDU\ YLWDPLQ ( LQFUHDVHG VHUXP DQG WLVVXH WRFRSKHURO FRQFHQWUDWLRQV 9LWDPLQ $ LQMHFWLRQV KDG OLWWOH RU QR

PAGE 71

HIIHFW RQ WKHVH FRQFHQWUDWLRQV GXULQJ HDUO\ JHVWDWLRQ RI JLOWV

PAGE 72

&+$37(5 *(1(5$/ &21&/86,216 7KUHH H[SHULPHQWV ZHUH FRQGXFWHG RQH WR GHWHUPLQH WKH ELRDYDLODELOLW\ RI IRXU IRUPV RI YLWDPLQ ( FRPSRXQGV DQG WZR WR DVVHVV WKH HIIHFW RI KLJK OHYHOV RI YLWDPLQ $ RQ WKH YLWDPLQ ( VWDWXV RI JURZLQJ ILQLVKLQJ SLJV RU JHVWDWLQJ JLOWV ,Q H[SHULPHQW WKH ELRSRWHQF\ RI IRXU IRUPV RI YLWDPLQ ( ZHUH GHWHUPLQHG *HQHUDOO\ WKH DFHWDWH IRUPV UHVXOWHG LQ JUHDWHU VHUXP DQG WLVVXH FRQFHQWUDWLRQV RI YLWDPLQ ( D WRFRSKHUROf WKDQ WKH DOFRKRO IRUPV GXH WR WKH JUHDWHU VWDELOLW\ RI WKH DFHWDWH IRUPV WKDW ZDV QRWHG LQ PL[HG IHHG 6HUXP WRFRSKHURO LQFUHDVHG UDWKHU UDSLGO\ ZKHQ WKH IRXU FRPSRXQGV ZHUH IHG 'LHWDU\ VXSSOHPHQWDWLRQ RI 'D WRFRSKHU\O DFHWDWH UHVXOWHG LQ WKH KLJKHVW VHUXP WRFRSKHURO WKURXJKRXW WKH VWXG\ FRPSDUHG WR FRQFHQWUDWLRQV REWDLQHG IRU SLJV IHG WKH RWKHU FRPSRXQGV LQGLFDWLQJ D JUHDWHU ELRSRWHQF\ ,8PJf IRU VZLQH WKDQ GHWHUPLQHG E\ WKH WUDGLWLRQDO UDW ELRDVVD\ $ VLPLODU WUHQG ZDV REVHUYHG ZLWK WLVVXH OLYHU EDFN IDW OHDI IDW DQG PXVFOHf WRFRSKHURO FRQFHQWUDWLRQV DV ZLWK VHUXP FRQFHQWUDWLRQV ZLWK WKH OLYHU KDYLQJ WKH KLJKHVW FRQFHQWUDWLRQ ,Q JHQHUDO DOO IRUPV ZRXOG SUREDEO\ EH VXLWDEOH GLHWDU\ VXSSOHPHQWDO VRXUFHV LI

PAGE 73

WKH VWDELOLW\ RI WKH DOFRKROV ZHUH LPSURYHG (QFDSVXODWLQJ WKH DOFRKRO IRUPV WR SURWHFW WKHP IURP GHVWUXFWLRQ ZRXOG LQFUHDVH WKHLU VXLWDELOLW\ IRU XVH LQ PL[HG IHHG ([SHULPHQW ZDV FRQGXFWHG WR HYDOXDWH WKH HIIHFW RI IHHGLQJ H[FHVVLYH YLWDPLQ $ RQ JURZWK SHUIRUPDQFH DQG RQ EORRG DQG WLVVXH DWRFRSKHURO YLWDPLQ (f OHYHOV RI JURZLQJn ILQLVKLQJ SLJV +LJK GLHWDU\ YLWDPLQ $ ,8NJ RI GLHWf ZDV IRXQG QRW WR DIIHFW RU KDYH OLWWOH DIIHFW RQ SLJ SHUIRUPDQFH RU RQ EORRG RU WLVVXH FRQFHQWUDWLRQV RI D WRFRSKHURO $ WKUHHIROG LQFUHDVH 3 f LQ VHUXP WRFRSKHURO RFFXUUHG RQ DOO VDPSOLQJ GD\V ZKHQ GLHWDU\ VXSSOHPHQWDWLRQ LQFUHDVHG IURP WR ,8NJ 7LVVXH WRFRSKHURO DOVR LQFUHDVHG 3 f DV GLHWDU\ YLWDPLQ ( LQFUHDVHG IURP WR ,8NJ 7LVVXH WRFRSKHURO FRQFHQWUDWLRQ LQFUHDVHG 3 f E\ D IDFWRU RI DW OHDVW WZR LQ DOO WLVVXHV HYDOXDWHG /LYHU UHWLQRO LQFUHDVHG 3 f HLJKWIROG ZLWK D WHQIROG LQFUHDVH LQ GLHWDU\ YLWDPLQ $ (YHQ LQ WKH OLYHU ZKHQ YLWDPLQ $ UHWLQROf FRQFHQWUDWLRQ ZDV KLJK DWRFRSKHURO FRQFHQWUDWLRQ ZDV QRW DIIHFWHG 7KXV WKH IRUP RI YLWDPLQ $ ZLWKLQ WKH OLYHU LV QRW LQ D IRUP ZKLFK FDQ OHDG WR R[LGDWLYH GHVWUXFWLRQ RI DWRFRSKHURO RU WKH FRQFHQWUDWLRQV HQFRXQWHUHG PD\ QRW KDYH EHHQ KLJK HQRXJK WR DIIHFW DWRFRSKHURO ([SHULPHQW HYDOXDWHG WKH HIIHFW RI JLYLQJ D KLJK OHYHO RI YLWDPLQ $ YLD LQWUDPXVFXODU LQMHFWLRQV RQ UHSURGXFWLYH SHUIRUPDQFH DQG RQ VHUXP DQG WLVVXH D

PAGE 74

WRFRSKHURO FRQFHQWUDWLRQV GXULQJ HDUO\ JHVWDWLRQ RI JLOWV +LJK OHYHOV RI YLWDPLQ $ ,8 SHU ZHHNf GLG QRW DIIHFW UHSURGXFWLYH SHUIRUPDQFH RU VHUXP RU WLVVXH FRQFHQWUDWLRQV RI DWRFRSKHURO RU UHWLQRO LQ WKLV VWXG\ $V REVHUYHG LQ WKH SUHYLRXV VWXGLHV LQFUHDVLQJ GLHWDU\ OHYHOV RI YLWDPLQ ( LQFUHDVHG EORRG VHUXP DQG WLVVXH WRFRSKHURO FRQFHQWUDWLRQV LQFOXGLQJ UHSURGXFWLYH WLVVXHV $OSKD WRFRSKHURO FRQFHQWUDWLRQ DOVR LQFUHDVHG LQ WKH HPEU\R ZKHQ GLHWDU\ YLWDPLQ ( ZDV LQFUHDVHG 7KH LQFUHDVHG FRQFHQWUDWLRQ RI DWRFRSKHURO LQGLFDWHV D WUDQVIHU RI WRFRSKHURO IURP WKH GDP WR WKH GHYHORSLQJ HPEU\R )OXFWXDWLRQV RI UHWLQRO FRQFHQWUDWLRQV LQ VHUXP PD\ KDYH EHHQ PLVVHG GXH WR ZHHNO\ VDPSOLQJ RI EORRG %ORRG VDPSOLQJ RQ G RU RQ D PRUH IUHTXHQW URXWLQH DIWHU YLWDPLQ $ LQMHFWLRQV ZRXOG SURYLGH PRUH DQVZHUV )XUWKHU UHVHDUFK ZRXOG EH GHVLUDEOH WR GHWHUPLQH YLWDPLQ ( ELRDYDLODELOLW\ RWKHU WKDQ E\ RUDO DGPLQLVWUDWLRQ )HFDO VDPSOH WRFRSKHURO DQDO\VLV PD\ SURYLGH PRUH LQIRUPDWLRQ IURP ZKLFK EHWWHU FRQFOXVLRQV PLJKW EH GUDZQ FRQFHUQLQJ GLJHVWLRQ DQG DEVRUSWLRQ 6WXGLHV XVLQJ ODUJHU QXPEHUV RI JLOWV DQGRU VRZV LV UHFRPPHQGHG WR GHWHUPLQH LI YLWDPLQ $ LQMHFWLRQ DORQH RU LQ FRPELQDWLRQ ZLWK KLJK YLWDPLQ ( HLWKHU RUDOO\ RU E\ LQMHFWLRQ ZRXOG HQKDQFH UHSURGXFWLYH SHUIRUPDQFH

PAGE 75

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

PAGE 76

H[FHVV YLWDPLQ $ DOWHU DWRFRSKHURO FDURWHQRLG DQG LURQ VWDWXV LQ DGXOW UDWV 1XWU %UDQGQHU 9LWDPLQ ( %HOO 3XEOLVKLQJ &RPSDQ\ 1HZ
PAGE 77

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

PAGE 78

PXVFOH WLVVXHV LQ WKH SLJ GXULQJ GHSOHWLRQ DQG UHSOHWLRQ $FWD 9HW 6FDQG .DQHNR &OLQLFDO %LRFKHPLVWU\ RI 'RPHVWLF $QLPDOV UG (Gf $FDGHPLF 3UHVV 1HZ
PAGE 79

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f 1DWLRQDO $FDGHP\ 3UHVV :DVKLQJWRQ '& 3XGHONLHZLF] : / :HEVWHU DQG / 0DWWHUVRQ (IIHFWV RI KLJK OHYHOV RI GLHWDU\ YLWDPLQ $ DFHWDWH RQ WLVVXH WRFRSKHURO DQG VRPH UHODWHG REVHUYDWLRQV 1XWU 5DDFNH +HUEHUW 0F/HDQ (YDQV D ELRJUDSKLFDO VNHWFK 1XWU f 6$6 6$6 8VHUnV *XLGH 6WDWLVWLFV 6$6 ,QVW ,QF &DU\ 1& 6FRWW 0 / 6WXGLHV RI YLWDPLQ ( DQG UHODWHG IDFWRUV LQ QXWULWLRQ DQG PHWDEROLVP ,Q 'H/XFD + ) DQG : 6XWWLH 7KH )DW 6ROXEOH 9LWDPLQV 7KH 8QLYHUVLW\ RI :LVFRQVLQ 3UHVV 0DGLVRQ 6NODQ DQG 6 'RQRJKXH 9LWDPLQ ( UHVSRQVH WR KLJK GLHWDU\ YLWDPLQ $ LQ WKH FKLFN 1XWU 7DSSHO $ / 9LWDPLQ ( DV WKH ELRORJLFDO OLSLG DQWLR[LGDQW 9LWDPLQV DQG +RUPRQHV 7KH $PHULFDQ +HULWDJH 'LFWLRQDU\ QG (Gf +RXJKWRQ 0LIIOLQ %RVWRQ 7KRPSVRQ 9LWDPLQ ( LQMHFWLRQV LPSURYH SLJ VXUYLYDO )HHGVWXIIV f 8OOUH\ ( 9LWDPLQ ( IRU VZLQH $QLP 6FL :HDYHU ( 0 : /LEDO & 5 +DPLOWRQ DQG 6 3DUNHU 5HODWLRQVKLS EHWZHHQ GLHWDU\ YLWDPLQ $ DQG ( RQ SHUIRUPDQFH DQG YLWDPLQ ( VWDWXV RI WKH ZHDQHG SLJ $QLP 6FL 6XSSO f $EVWUf

PAGE 80

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

PAGE 81

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n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

PAGE 82

7KLV GLVVHUWDWLRQ ZDV VXEPLWWHG WR WKH *UDGXDWH )DFXOW\ RI WKH &ROOHJH RI $JULFXOWXUH DQG WR WKH *UDGXDWH 6FKRRO DQG ZDV DFFHSWHG DV SDUWLDO IXOILOOPHQW RI WKH UHJXLUHPHQWV IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ $XJXVW 'HDQ *UDGXDWH 6FKRRO


xml version 1.0 encoding UTF-8
REPORT xmlns http:www.fcla.edudlsmddaitss xmlns:xsi http:www.w3.org2001XMLSchema-instance xsi:schemaLocation http:www.fcla.edudlsmddaitssdaitssReport.xsd
INGEST IEID EBI3B40QP_4J16UB INGEST_TIME 2011-11-08T18:11:36Z PACKAGE AA00004732_00001
AGREEMENT_INFO ACCOUNT UF PROJECT UFDC
FILES