Citation
Mineral status of sheep in the Paramo region of Colombia

Material Information

Title:
Mineral status of sheep in the Paramo region of Colombia
Creator:
Pastrana, Rodrigo, 1945-
Publication Date:
Language:
English
Physical Description:
xii, 192 leaves : maps ; 29 cm.

Subjects

Subjects / Keywords:
Calcium ( jstor )
Dry seasons ( jstor )
Forage ( jstor )
Magnesium ( jstor )
Minerals ( jstor )
Phosphorus ( jstor )
Rainy seasons ( jstor )
Sheep ( jstor )
Soils ( jstor )
Zinc ( jstor )
Animal Science thesis Ph. D
Dissertations, Academic -- Animal Science -- UF
City of Gainesville ( local )
Genre:
bibliography ( marcgt )
theses ( marcgt )
non-fiction ( marcgt )

Notes

Thesis:
Thesis (Ph. D.)--University of Florida, 1989.
Bibliography:
Includes bibliographical references (leaves 180-191).
Additional Physical Form:
Also available online.
General Note:
Typescript.
General Note:
Vita.
Statement of Responsibility:
by Rodrigo Pastrana.

Record Information

Source Institution:
University of Florida
Holding Location:
University of Florida
Rights Management:
Copyright Rodrigo Pastrana. 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:
22655259 ( OCLC )
022586630 ( ALEPH )

Downloads

This item has the following downloads:


Full Text














MINERAL STATUS OF SHEEP IN THE
PARAMO REGION OF COLOMBIA















BY

RODRIGO PASTRANA














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 1989











ACKNOWLEDGMENTS


I wish to express my sincere gratitude and appreciation to Dr. Lee R. McDowell, adviser and chairman of my supervisory committee, for his valuable guidance and assistance throughout the investigation and preparation of the present dissertation. Acknowledgments are also due to to Dr. Joseph H. Conrad, Clarence B. Ammerman, Douglas B. Bates and Lynn E. Sollenberger for their time and advice as members of the supervisory committee.

Recognition and appreciation are due to Mrs. Nancy

Wilkinson and Mr. Richard Fethiere for their assistance in all laboratory work and to Dr. Frank G. Martin and Dr. Steve Linda for their assistance in statistical analysis.

Special appreciation is due to Dr. Oliver Ospina of Caja Agraria and to Dr. Alfonso Naranjo of Instituto Colombiano Agropecuario (ICA), who permitted the sample collection in order to maIe this work possible, to Dr. Rodrigo Lora for soil analysis and to Dr. Jorge Neira for whole blood analysis at ICA laboratories.

Special recognition is due to Instituto Colombiano

Agropecuario for the financial support of my studies in the United States.










Deep appreciation goes to my wife Diana and to my five children for their companionship and encouragement during this hardship. To them, this dissertation is gratefully dedicated.












TABLE OF CONTENTS


ACKNOWLEDGMENTS .........

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

LIST OF FIGURES .........

ABSTRACT ............

CHAPTER

I INTRODUCTION .......

II LITERATURE REVIEW

Characterization of the

Mineral Status of Soil
Soil Acidity and 0 Soil Macronutrient Soil Micronutrient
Mineral Status of Plant
Forage Macrominera Forage Microminera
Mineral Status and Requ
Tissue Macrominera Tissue Microminera
Hematological Measureme

III MATERIALS AND METHODS.

Identification and Desc
Sample Collection . .
Soil Samples. . .
Forage Samples. .
Animal Tissue Samp
Whole blood a
Bone biopsy
Liver biopsy


Page

. . . . . . . . . . . . . . ii
. . . . . . . . . . . . . . vii



. . . . . . . . . . . . . . xi



� . �. ,o. o . . . . . . . . 1



Sheep Industry in Colombia 3
. . . . . . . . . . . . . . 7
rganic Matter ... ....... 7
s ...... ............. 10
s .... ............. .. 12
s ..... ............. 16
is .... ............. .. 18
is .... ............. .. 20
irements of Ruminants . 23 is .... ............. .. 23
is .... ............. .. 29
nts .... ............ 33

. . . . . . . . . . . . . . 36

ription of Research . . . . 36 . . . . . . . . . . . . . . 37
. . . . . . . . . . . . . . 37
. . . . . . . . . .. . . . . 39
les .... ............ 40


nd blood serum











Chemical Analysis ....... ................
Soil Samples ....... ................
Forage Samples ...... ................
Animal Tissue Samples .... ............
Blood serum .............
Whole blood ...... ..............
Bone sample ...... ..............
Liver samples ..... ..............
Statistical Analysis ...... ..............

IV MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF
COLOMBIA: I. MACROELEMENTS .... ...........


Introduction .... ...........
Materials and Methods ......... Results and Discussion .......
Soil Analyses .. ........
Forage Analyses ........
Animal Tissue Analyses . .
Blood serum .....
Bone ... ..........
Correlation Coefficients of Hematological Measurements. Summary and Conclusions ........


Minerals ........


V MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF
COLOMBIA: II. MICROELEMENTS ..... .............


Introduction .... ...........
Materials and Methods ......... Results and Discussion .......
Soil Analyses .. ........
Forage Analyses ........
Animal Tissue Analyses . .
Blood serum .....
Liver . . .. .. .
Correlation Coefficients of Summary and Conclusions .......


Minerals.


VI MINERAL CONCENTRATIONS IN LEAVES AND STEMS OF
VARIOUS FORAGES OF THE COLOMBIAN PARAMO .........

Introduction ........ ...................
Materials and Methods ...... ...............


Pace

43
43 44 45
45
* 45
* 46
* 46
* 48


* 49









Page


Results and Discussion ..... .............. 96
Plant Fractions-Macrominerals, Crude Protein,


and IVOMD ... ..........


Plant Fractions-Microminerals .
Forage Species ... ..........
Soil-Plant Relationship ....
Summary and Conclusions .......... VII SUMMARY AND CONCLUSIONS .......... APPENDIX A FIGURES .... ............

APPENDIX B TABLES ..... .............

APPENDIX C RAW DATA ... .............

LITERATURE CITED .... ...............

BIOGRAPHICAL SKETCH .... ............


. . . . . . . 96
. . . . . . . 99
. . . . . . . 102
. . . . . . . 108
. . . . . . . 110

. . . . . . . 112

. . . . . . . 117

. . . . . . . 121

. . . . . . . 146

. . . . . . . 180

. . . . . . . 192


o
Q













LIST OF TABLES


Table Paae

1. CONCENTRATIONS (PPM) OF MINERALS OF DRIED SOIL
AND THEIR INTERPRETATION .... ............. 13

2. SUMMARY GUIDE TO MINERAL REQUIREMENTS FOR RUMINANTS
(DRY BASIS) ......... .................... 17

3. DIAGNOSIS OF SPECIFIC MINERAL DEFICIENCIES OR
TOXICITIES IN SHEEP ...... ............... 24

4. NORMAL BLOOD HEMOGLOBIN, HEMATOCRIT AND LEUCOCYTE
VALUES IN SHEEP ....... ................. 35

5. SOIL ORGANIC MATTER, pH, AND MACROMINERAL ANALYSES
AS RELATED TO SEASON AND FARM (DRY BASIS) . . . . 54

6. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOMD
CONCENTRATIONS AS RELATED TO SEASON AND FARM (DRY
BASIS) ......... ...................... 56

7. BLOOD SERUM AND BONE MACROMINERAL CONCENTRATIONS AS
RELATED TO SEASON AND FARM ..... ............. 61

8. BLOOD SERUM AND BONE MACROMINERAL CONCENTRATIONS AS
RELATED TO SEASON AND ANIMAL CLASS ........... . 62

9. HEMOGLOBIN, HEMATOCRIT CONCENTRATIONS AND LEUCOCYTE
COUNTS AS RELATED TO SEASON AND FARM .......... .. 68

10. SOIL MICROMINERAL ANALYSES AS RELATED TO SEASON AND
FARM DRY BASIS) ....... .................. 76

11. FORAGE MICROMINERAL CONCENTRATIONS AS RELATED TO
SEASON AND FARM (DRY BASIS) .................. 79

12. BLOOD SERUM AND LIVER MICROMINERAL CONCENTRATIONS
AS RELATED TO SEASON AND FARM ... ........... 83

13. BLOOD SERUM AND LIVER MICROMINERAL CONCENTRATIONS
AS RELATED TO SEASON AND ANIMAL CLASS ......... . 84


vii










Table Page

14. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOMD
CONCENTRATIONS AS RELATED TO SEASON AND PLANT PART
(DRY BASIS) ......... .................... 98

15. FORAGE MICROMINERAL CONCENTRATIONS AS RELATED TO
SEASON AND PLANT PART (DRY BASIS) ........... . 100

16. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOMD
CONCENTRATIONS BY SPECIES (DRY BASIS) ......... . 103 17. FORAGE MICROMINERAL CONCENTRATIONS BY SPECIES
(DRY BASIS) ........ .................... 104

18. CORRELATION COEFFICIENTS BETWEEN SOIL AND FORAGE
MINERALS AS RELATED TO SEASON ... .......... 109

19. DESCRIPTION OF FARMS ...... ............... 121

20. CLIMATE AND AVERAGE MONTHLY RAINFALL (MM) ..... . 123 21. DETAILED NUMBER OF SOIL AND FORAGE COMPOSITE
SAMPLES ......... ...................... 124

22. COMPOSITION OF THE MINERAL MIXTURES .......... . 125

23. DETAILED NUMBER OF TISSUE SAMPLES ........... . 126

24. SUMMARY OF ANALYSES OF VARIANCE OF SOIL ORGANIC
MATTER, PH, AND MACROMINERALS-MEAN SQUARES BY
SEASON ......... ...................... 127

25. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE CRUDE
PROTEINIVOMD, AND MICROMINERALS-MEAN SQUARES
BY SEASON ......... .................... 128

26. SUMMARY OF ANALYSES OF VARIANCE OF BLOOD SERUM
AND BONE MACROMINERALS-MEAN SQUARES BY SEASON. . . 129

27. SUMMARY OF ANALYSIS OF VARIANCE OF WHOLE BLOOD
VARIABLES--MEAN SQUARES BY SEASON ........... . 130

28. BLOOD SERUM AND BONE MACROMINERAL CORRELATION
COEFFICIENTS AS RELATED TO SEASON ........... . 131

29. CORRELATION COEFFICIENTS BETWEEN BLOOD SERUM,
LIVER, AND BONE MINERALS AS RELATED TO SEASON. . . 132








Table Pacre

30. SUMMARY OF ANALYSES OF VARIANCE OF SOIL
MICROMINERALS-MEAN SQUARES BY SEASON ........ . 133

31. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE
MICROMINERALS--MEAN SQUARES BY SEASON . . . . 134

32. SUMMARY OF ANALYSES OF VARIANCE OF BLOOD SERUM
AND LIVER MICROMINERALS-MEAN SQUARES BY SEASON . . 135

33. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE (PARTS),
CRUDE PROTEIN, IVOMD, AND MACROMINERALS-MEAN
SQUARES BY SEASON ...... ................. 136

34. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE (PARTS)
MICROMINERALS-MEAN SQUARES BY SEASON ........ . 137

35. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOND
CONCENTRATIONS AS RELATED TO SEASON AND SPECIES
(DRY BASIS) ........ .................... 138

36. FORAGE MICROMINERAL CONCENTRATIONS AS RELATED TO
SEASON AND SPECIES (DRY BASIS) .. .......... 140

37. OVERALL FORAGE MINERAL CONCENTRATIONS IN DIFFERENT
SPECIES (DRY BASIS) ..... ............... 142

38. SOIL ORGANIC MATTER, PH, MACROMINERAL AND
MICROMINERAL CORRELATION COEFFICIENTS AS RELATED
TO SEASON ......... ..................... 143

39. FORAGE MACROMINERAL, MICROMINERAL, CRUDE PROTEIN,
AND IVOMD CORRELATION COEFFICIENTS AS RELATED TO
SEASON ......... ...................... 144













LIST OF FIGURES


FPage


1. COLOMBIA, GEOGRAPHICAL LOCATION OF THE THREE SHEEP
FARMS SURVEYED ....... .................. 117

2. THE PARANO REGION IN COLOMBIA ... ........... 118













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

MINERAL STATUS OF SHEEP IN THE
PARAMO REGION OF COLOMBIA

By

RODRIGO PASTRANA

December 1989

Chairman: Dr. Lee R. McDowell Major Department: Animal Science

A study was conducted in the paramo region of Colombia to determine the mineral status of grazing sheep by evaluating mineral concentrations in soil, plant, and animal tissues. Samples were taken at the end of the rainy season (May-June, 1987) and at the end of the dry season (February, 1988) from lactating (or pregnant) ewes, lambs, and yearlings. A total of 113 composite soil, 131 composite forage, 207 blood serum, 192 whole blood, 148 bone rib biopsy, and 113 liver biopsy samples were obtained.

Percentage of soil samples that were deficient was

selenium 100%, boron 83%, magnesium 70%, manganese 54%, and potassium 52%. Percentage of forage samples that were deficient was sodium 93%, cobalt 87%, copper 75%, selenium 56%, magnesium 53%, and crude protein 6%. None of the values for molybdenum were above the toxic limit of 6 ppm for sheep, and the copper:molybdenum ratio was at least 4:1.







Iron and manganese were in excess of the requirements but below the maximum tolerable levels for sheep. Leaves had higher (P<.05) concentrations of calcium, phosphorus, magnesium, potassium, iron, molybdenum and crude protein than stems. Among soil minerals and the corresponding forage minerals, only calcium and magnesium had positive correlation coefficients (P<.05, r>1.501) for both seasons.

Serum showed phosphorus deficiency in 59% of the

samples during both seasons, and calcium 94% during the dry season. Both calcium and phosphorus were deficient in 98% of bone samples. Liver indicated a copper deficiency in 59% of the samples, and of zinc in 44%; during the dry season 55% were deficient in cobalt. Differences (P<.05) among animal classes were found in serum phosphorus in both seasons as was the case for serum magnesium during the dry season and bone ash during the rainy season.

In conclusion, minerals most likely limiting sheep production in the paramo were calcium, phosphorus, magnesium, copper, zinc and cobalt. Supplementation programs should provide the above minerals plus common salt and selenium.


xii














CHAPTER I
INTRODUCTION


Paramo is an ecological concept that refers to the

mountainous regions of the humid equatorial Andes above the upper limit of the forest (3,100 m).. It is characterized by extreme environmental conditions, acid soil, low atmospheric pressure, low mean temperature, and high humidity. As such, it is an unique phenomenom in our planet, which only occurs in four countries: Colombia, Venezuela, Ecuador, and Costa Rica. The great importance of the paramo in Colombia is that this region is the country's source of water; most of the rivers have their origin in these very high lands. The paramo is covered with shrubs and native grasses, and is better suited for sheep grazing rather than cattle.

About 47% of the sheep population (total population: 2,464,000) are wooled sheep that graze in the mountains. However, sheep production is not very efficient. Feed is in short supply during the dry season, and in general the few forage species that grow in the paramo are considered of low quality and their regrowth is very slow after grazing. Under these conditions, the animals do not consume their nutritional requirements and produce inefficiently. The poor growth rate of lambs, low fertility in particular of





2

imported breeds, high mortality, and low wool production are characteristics of sheep in the paramo of Colombia.

As is known, ruminants grazing forages in severely mineral-deficient areas may be more limited by mineral insufficiency than either a lack of protein or energy. Mineral deficiencies or imbalances in soils and forages are often responsible for low production and reproduction among grazing livestock. As grazing livestock usually do not receive mineral supplementation, except for common salt, they must depend almost exclusively upon forages for their requirements. Therefore, supplemental feeding of adequate minerals, which are frequently deficient in paramo forages, may improve this problem.

The objectives of this study were as follows:

1. to establish if specific mineral deficiencies or toxicities exist in the paramo region of Colombia;

2. to compare the seasonal (rainy vs dry) mineral status of soil, forages, and sheep tissues;

3. to evaluate animal tissues at different physiological states of the sheep (lactating-pregnant ewes, lambs, and yearlings); and

4. to study mineral soil-plant-animal interrelationships.















CHAPTER II
LITERATURE REVIEW

Characterization of the Sheen Industry in Colombia


Colombia, the northernmost of all South American republics, is located at the north east corner of South America between 40 50' to 790 01' west of the Greenwich Meridian (Instituto Geografico Agustin Codazzi, 1986). The capital of Colombia, Bogota, lies at an altitude of 2,640 m above sea level (Appendix A, Figure 1).

The total area of the country is 1,141,748 Km2 and, while there is a great variety of geographical areas, the country has been divided in 5 natural regions. The Andes region and its interandean valleys include the high mountain ranges of the three Cordillera chains (Cordilleras Oriental, Central and Occidental), the Macizo de los Andes and the Sierra Nevada. The Coastal plains, both Pacific and Atlantic, and the Savanna (Llanos Orientales) are included in the Llanuras del Pacifico, Llanuras del Caribe, and the Orinoquia regions, respectively. A dry, almost desert zone in the Peninsula de la Guajira is also included in the Caribe region. A tropical jungle is found in the Amazonia region (Instituto Colombiano Agropecuario, 1980).







In general, two different types of sheep, hair and wooled, are found in Colombia, according to two climatic regions. The hair sheep are concentrated in the lowlands with high temperatures all year, which range from 20 to 330C under both humid and very dry conditions. Hair sheep are well adapted to tropical environments. The African sheep, as hair sheep are known in Colombia, are usually brown, ranging in shade from tan to brown and cherry-red to dark red (Bradford and Fitzhugh, 1983).

The Peninsula de la Guajira holds about 30% of the

total sheep population, most of which are hair sheep. The total sheep numbers in the country were estimated to be 2,464,000 in 1982 (Caja Agraria, 1985).

The wooled sheep are found in the mountain regions

where temperatures range from 9 to 150 C; in most cases they graze the higher elevations of the mountains, called paramos, where temperatures are lower than 90C and humid conditions persist. The departamento de Boyaca leads the country in wooled sheep numbers with about 26% of the total sheep population, followed by Cundinamarca with close to 10%. Santander and Narino are also important regions and they both have 11% of the sheep in Colombia (Caja Agraria, 1985).

The predominant type of wooled sheep is the Native or Criolla breed which is well adapted to the harsh environment of the paramo. Importations of Romney, Corriedale, Hampshire, Scottish Blackface and others have occurred in







the last 30 years and substantial crossing of the Criolla has resulted especially with the Romney breed.

In general, the paramo region has not been exploited in a systematic way and its utilization is in its infancy. The production capability of a sheep industry in the paramo is difficult to determine as sheep have to compete not only with technological but also social restrictions, for example, lack of roads, isolation, lack of electricity, and others (Proyecto Ovino Colombo Britanico, 1979). Among the technical restrictions to the sheep industry in the paramo we can mention the poor growth rate of the lambs until weaning; low bodyweights at 6, 12, and 18 months of age; low fertility in particular with imported breeds; high mortality rate due to several causes; and low production of wool of inferior quality. One important restriction is the very low production of forage which can be attributed to cold weather and periods of excess rainfall or drought (Proyecto Ovino Colombo Britanico, 1979).

In a description of the region, Guhl (1982) contends that the paramo is an ecological concept that refers to the mountainous regions of the humid equatorial Andes above the upper limit of the forest (above 3,100 m). The paramo is characterized by extreme environmental conditions, acid soils, low atmospheric pressure, high humidity, low mean temperature (with daily oscilation), and high air dryness. It is also characterized by the elevated soil and air temperatures during the direct solar radiation and by the






6
sudden and dramatic changes due to the cloudiness during the night. These factors produce frosts and heavy winds during specific times of the year.

The paramo is a unique phenomenom in our planet, which only occurs in four countries, Colombia, Venezuela, and Ecuador. In Costa Rica the climatic effect of elevations at 2,500 m above sea level are comparable to those of the paramos (Robayo et al., 1988). Very little is yet known about the complex interrelationships that keep animal and plant life and that permit the existence of the paramos. Less is known of the possible utilization of the paramo lands by man.

The landscape in the paramo is not totally homogeneous, and three altitudinal zones may be distinguished: the superparamo on the higher elevations (above 4,200 m), the proper paramo (3,800-4,200 m, and the subparamo on the lower elevations (3,100-3,800 m) (Salamanca, 1986). The paramo region can be observed in appendix A figure 2.

The natural vegetation in the paramo is adapted to

withstand cold and dry conditions. Although the paramo is very humid, in some cases the effective rainfall is low (less than 1,000 mm in a year). A permanent fog in combination with low temperatures keeps the relative humidity at an almost 100%. The soils are generally saturated but is difficult for the plants to utilize the water and so they are in a similar situation as if they were in an arid zone (Salamanca, 1986).







Apart from its possible utilization with grazing

animals or other forms of agriculture, the great ecological importance of the paramo lies as a source of water. Most of the rivers in Colombia have their origin in these very high lands (Cleef, 1980).

Around 70% of the Colombian population live and work on the mountains. But the mountain has several aspects, and the higher elevations, as are the paramos, become a hostile region to man for its climate and topography, and are natural obstacles to communications (Guhl, 1982).



Mineral Status of Soil


Soil Acidity and Organic Matter

McDowell (1985) indicates that soil is the source of all mineral elements found in plants. Most naturally occurring deficiencies in livestock are associated with specific regions and are directly related to both soil mineral concentration and soil characteristics. Of the total mineral concentration in soils, only a fraction is taken up by plants.

Conrad (1978) indicates that grazing ruminants are in close and continuing contact with the soils on which the plants grow. Soils often contain many times the levels of minerals which are found in the plants which grow on these soils.







For Reid and Horvath (1980), the availability of

minerals in soils depends upon their effective concentration in soil solution. Williams (1963) indicates that the soil content of an element would seem the most important limitation. However, availability factors including soil pH, texture, moisture content, and organic matter are probably more often the limiting factors rather than soil mineral content.

Sanchez (1976) says that the majority of the soils of the humid tropics is acid. Soil acidity problems are associated with pH levels lower than 5.5 and the presence of exchangeable aluminum in the soil. Percent aluminum saturation, calculated on the basis of effective cation exchange capacity, is a useful measure of soil acidity.

For Sanchez (1976), acid soil infertility is due to one or more of the following factors: aluminum toxicity, calcium or magnesium deficiency, and manganese toxicity. Aluminum toxicity is the most common cause of acid soil infertility. This toxicity can be corrected by liming to pH 5.5 to 6.0 to precipitate the exchangeable aluminum as aluminum hydroxide. Liming rates can be calculated on the basis of 1.65 ton/ha of CaCO3 equivalent per milliequivalent of exchangeable aluminum. Overliming to pH values greater than 6 or 7 can seriously decrease yields, particularly in soils high in iron and aluminum oxides (Kamprath, 1972).

Concentrations of soil solution aluminum above 1 ppm often causes direct reduction in yield. Aluminum tends to







accumulate in the roots and impede the uptake and translocation of calcium and phosphorus. Thus aluminum toxicity may produce or accentuate calcium and phosphorus deficiencies (Foy, 1974).

Under grazing conditions, sheep and cattle can ingest large amounts of aluminum of nonplant origin when soil is involuntarily ingested with forages. Exchangeable aluminum concentrations of 6,000 and 18,000 ppm have been reported for samples of temperate and tropical soils, respectively (Velez and Blue, 1971).

Manganese is very soluble at pH values lower than 5.5. If present in sufficient amounts, manganese toxicity can occur along with aluminum toxicity at pH values of up to about 5.5 to 6.0. Manganese is a plant nutrient; consequently, the aim is not to eliminate soluble manganese but to keep it within a range between toxicity and deficiency. A solution concentration between 1 and 4 ppm represents such a range (Black, 1967).

Referring to organic matter, Sanchez (1976) indicates that in unfertilized soils the beneficial effects of organic matter consist of supplying most of the nitrogen and sulfur to plants, maintaining cation exchange capacity, blocking phosphorus fixation sites, improving structure in poorly aggregated soils, and the formation of complexes with micronutrients. For Dahnke and Vasey (1973), a test for total nitrogen and a test for organic matter or organic







carbon are essentially the same because the concentration of nitrogen in soil organic matter is relatively constant. Soil Macronutrients

Towers and Clark (1983) indicated that mineral

concentrations in both soil and plant influence the mineral status of grazing animals but many other factors such as total dry matter intake, selective grazing, interactions among minerals, variations in mineral requirements with differences in age, sex or production level also play an important part. This means that soil analysis alone cannot be used to reliably diagnose trace element deficiencies in animals.

A more satisfactory soil analysis to relate mineral concentrations for livestock rather than the concentration of a mineral in a soil, is the use of soil extracts (i.e.,

0.1N HCI or 2.5% acetic acid), which contain the more available forms of soil minerals. Analyses to determine the available forms of soil minerals can sometimes provide evidence of livestock mineral deficiencies, but more often they are unreliable and difficult to interpret (McDowell et al., 1986).

Calcium is essential, not only to correct soil acidity but also as a nutrient element necessary for normal plant growth. Soil calcium plays an essential role in regulating soil pH (Sillanpaa, 1982). The same author determined that the calcium concentration of soils are poorly reflected in the calcium concentration of plants.







Doll and Lucas (1973) stated that plants usually contain more potassium than any other nutrient element except nitrogen. Crops utilize from 50 to over 200 kg/ha, depending upon the yield and kind of crop. Supplemental potassium fertilizers are frequently needed since most soils can not meet the requirements of continued cropping. Levels of calcium in crops are lower than potassium, and the amount needed by crops usually is between 20 and 150 kg Ca/ha. However, nutrient deficiencies of calcium are not common since most soils either contain high levels of calcium or have been supplied with ample calcium because of liming. Uptake of magnesium by crops is relatively low, from less than 10 to about 25 kg/ha. Occasional magnesium deficiencies have been noted, especially on sandy soils or soils with low levels of magnesium in relation to potassium and possibly calcium.

Phosphorus is classed as one of the macronutrients, but its concentration in plants is considerably less than that of nitrogen, potassium and calcium. As a limiting factor, however, phosphorus is more important than calcium and probably more important than potassium. The forms of phosphorus that occur in soil parent materials are generally of low availability to plants. Probably all forms of phosphorus in soils are of some significance in supplying phosphorus for plants on a long-term basis, but none of the forms that have been identified are known to be of significance in the short-term relationships that are of







importance in determining the current availability of soil phosphorus (Black, 1968).

Sanchez (1976) stated that in general, sulfur deficient soils have one or more of the following properties. They are high in allophane or oxides. They are also low in organic matter and often sandy. Soils subject to repeated annual burning are often sulfur deficient since about 75% of the sulfur is volatilized by fire. In temperate regions total soil sulfur is positively correlated with organic matter and inversely correlated with degree of weathering. Sanchez (1976) also stated that sulfur deficiency in grazing sheep and cattle is not a problem, but its effect on copper absorption and metabolism is important.

Sodium is of importance in plant nutrition not only because sodium is required by at least a few plants, but also because of its relation to potassium. Sodium and potassium are the two principal monovalent metallic cations in plants, and an increase in one generally brings about a decrease in the other (Black, 1968). Table 1 provides a summary guide to the levels of soil fertility and its interpretation according to several authors. Soil Micronutrients

Sulfur and the micronutrients can be differentiated from nitrogen, phosphorus, and potassium in that they are much less frequently the limiting factor in soil fertility (Foth and Ellis, 1988). Both copper and zinc occur in the earth's crust primarily as sulfide minerals. Deficiencies









TABLE 1. CONCENTRATIONS (PPM) OF MINERALS OF DRIED
INTERPRETATIONa


SOIL AND THEIR


Levels Soil
Element Low Medium High type Reference


Calcium 0-71 72-140 >141 b Magnesium 0-30 30-50 > 60 c Phosphorus 9-17 17-35 35-70 organic- c Mineral

Potassium 31-62 62-124 124-248 Organic- C Mineral

Sulfur <10 >10 d Boron < .4 e Copper < .1-.3 pH 5.5-6.0 c Iron < 2.5 > 4.5 f Manganese < 3-5 pH 5.5-6.0 c


Zinc < .5 pH 5.5-6.0 c


'For grazing livestock soil concentrations suggesting deficiencies are as follows: calcium (70 ppm), potassium (58.5 ppm), magnesium (8.4 ppm), phosphorus (10 ppm), cobalt (0.1 ppm), copper (0.6 ppm) ,manganese (19 ppm), and zinc (2 ppm) (McDowell et al., 1986). bBreland, 1976.

CRhue and Kidder, 1983.
dCooper, 1968.

eGammon, 1976.
fViets and Lindsay, 1973.







of copper are not commonly found in mineral soils. Organic soils containing little ash are more likely to be deficient. More than 99% of the copper in the soil solution is complexed by organic matter. This complexing is of great importance in maintaining adequate copper in solution for plant use. Organic matter does complex zinc in soil solution, but the percentage of zinc that is complexed varies over a considerable range (Foth and Ellis, 1988).

Since manganese solubility is related to oxidationreduction reactions in the soil, the availability of manganese is closely related to weather. Cool temperatures may slow down the mineralization of organic manganese. On the other hand, cool temperatures associated with high levels of rainfall in early spring may keep more manganese available through reduction of manganese oxides. There is an interaction between manganese and iron. High levels of available iron in organic soils or high levels of organic matter in sands may lead to a manganese deficiency because a high ratio of iron to manganese is created within the plant (Foth and Ellis, 1988).

Foth and Ellis (1988) also state that few, if any,

soils are deficient in total iron since the total soil iron concentration varies from 1,000 to 10,000 ppm. But the solubility of iron in soils may be limited by the low solubility of iron hydroxides and oxides in the pH range in which crops are grown. Soil conditions that lead to iron







deficiency in plants include pH above 7.0, low soil moisture concentration, and low organic matter concentration.

Boron is associated with soil organic matter, and soils with high levels of organic matter usually contain adequate boron for high soil fertility. Boron deficiency is often accentuated when soil contains little moisture.

As with other elements, the total concentration of selenium in soils shows little relationship to the concentration of selenium in plants grown on those soils. This is because selenium in soils exists in several chemical forms which differ widely in their solubility and availability to plants. The chemical forms of selenium (selenides, selenites, selenates, organic selenium) are closely related to oxidation-reduction potential and pH of the soil (Lakin, 1972).

Many studies have determined the factors affecting the availability of the microelements. The amount of most trace elements in herbage grown in freely drained soils is normally lower than on corresponding poorly drained soils (Swift, 1972).

As soil pH increases, the availability and uptake of iron, manganese, zinc, copper, and cobalt decrease, whereas molybdenum and selenium concentrations increase (Pfander, 1971; Williams, 1963; Latteur, 1962; Miller et al., 1972). Many data have accumulated indicating a decrease in the solubility of zinc with increasing pH (Foth and Ellis, 1988).







Low soil temperatures usually decrease micronutrient availability and may cause deficiency signs to develop during cold springs only to disappear when the soil warms up and more roots develop (Viets and Lindsay, 1973).



Mineral Status of Plants



Of the thirteen essential minerals obtained from the

soil by plants, five are used in relatively large quantities and are thus referred to as macronutrients. These are phosphorus, potassium, calcium, magnesium, and sulfur. The other minerals, iron, manganese, copper, zinc, boron, molybdenum, chlorine and cobalt, are used by higher plants in very small amounts, thereby giving them the designation of micronutrients or trace elements (Brady, 1974; McDowell et al., 1983).

Mineral analysis of the forage consumed by the grazing animal is basic to mineral status diagnosis. If mineral concentrations are below minimum requirements or above the maximum tolerance level, there is an immediate suggestion of a nutritional problem. However, relying on a forage mineral analysis to establish mineral status assumes that the sample is representative of what animals consume. An additional disadvantage of forage element analyses is the difficulty of estimating forage intake and digestibility (McDowell et al., 1986). Table 2 presents a guide to mineral element








TABLE 2. SUMMARY GUIDE TO MINERAL REQUIREMENTS FOR RUMINANTS
(DRY BASIS)



Element Requirementa Calcium .18-.60 % Phosphorus .18-.43 % Magnesium .04-.18 % Potassium .60-.80 % Sodium .10 % Iron 10-100 ppm Copper 4- 10 ppm Cobalt .05-.10 ppm Zinc 10- 50 ppm Manganese 20- 40 ppm Molybdenum .01 ppm or less "Summarized by McDowell et al. (1978).





18

requirements for grazing ruminants, dry basis, summarized by McDowell et al. (1978).

Gross and Jung (1981) established that forages supply much of the minerals in diets fed to cattle, and that variation in the amount of minerals in forages are associated with season, fertilization, soil type, and soil pH.
Both amount of mineral in forages and biological availability of minerals need to be considered in formulating rations. Although mineral concentration of forages can be determined chemically, biological availability is much more difficult to estimate. Biological availability of mineral elements in forages is probably a partial function of the extent to which they are solubilized in rumen fluid (Kincaid and Cronrath, 1983).

Grazing livestock usually do not receive mineral supplementation except for common salt and must depend almost exclusively upon forages to meet their requirements. Only rarely, however, can forages completely satisfy all mineral requirements (McDQwell et al., 1982). Forage Macrominerals

Phosphorus deficiency is found frequently-in tropical grazing areas around the world (Cohen, 1980), and may be the first limiting mineral deficiency under many grazing conditions. Aluminum and iron in ingested soil may interfere with dietary phosphorus utilization (Rosa et al.,







1982) and this effect could be critical if the animals were in a borderline phosphorus deficiency. The phosphorus requirement of a ruminant is rarely met by forage diets; therefore, supplementation is then necessary (Cohen, 1980).

Calcium deficiency is a lesser problem than phosphorus deficiency in grazing animals because most herbage contains adequate calcium, and maturity has only a small effect on calcium concentration. In contrast, most of the world's rangeland soils are low in phosphorus and support herbage of low phosphorus concentration which declines markedly with maturity (Underwood, 1981).

Magnesium occurs widely in plant and animal tissues. Magnesium content of most grazed herbages usually exceed

0.1% so requirements for supplemental magnesium in grazing cattle and sheep are usually associated with induced hypomagnesemia as a result of higher concentrations of nitrogen and potassium due to fertilization rather than low magnesium levels per se (Cohen, 1987). Metson et al. (1966) suggested that magnesium levels of 0.25% were necessary to prevent grass tetany when concentrations of nitrogen and potassium are high.

Potassium is often associated with high levels of

nitrogen and moisture in lush, cool-season grasses (Boling et al., 1979). Ruminants grazing pasture heavily fertilized with potassium have developed hypomagnesemic tetany, but adding potassium salts directly to their diet usually failed to produce this condition (Tomas and Potter, 1976).







Excess sodium salts in the diet may increase the rate of passage of digesta and, thus, enable a greater quantity of dietary protein to escape ruminal degradation (Reffett and Boling, 1985). Moseley and Jones (1974) reported that feeding high levels of NaCl resulted in increased magnesium absorption, but concomitantly resulted in increased urinary excretion of sodium and magnesium. Forage Microminerals

An accurate determination of zinc requirements of

ruminants is not available, although the level of 25 to 30 ppm in forage is consistent with results obtained from grazing experiments. Zinc concentrations may be 30 ppm in herbage and occasionaly higher, but this concentration declines rapidly as plants mature and values can decrease to less than 15 ppm (Mayland et al., 1987). The bioavailability of zinc may be reduced by cellulosic binding (Bremner and Knight, 1970).

Forage copper concentrations vary from 4 to 5 ppm to values of 10 to 15 ppm. Sulfur and molybdenum interfere with the absorption of copper. Variations in copper absorption may more often be associated with changes in soil pH or redox potential that have affected the solubility and uptake of molybdenum and/or sulfur (Langlands et al., 1981; Lesperance et al., 1985).

Absorption of copper varies according to age and

dietary factors such as the level of molybdenum and sulfur. Zinc and calcium levels in the diet may also interact with







copper absorption, but to a lesser extent than do molybdenum and sulfur. Herbage containing 5-6 and 7-10 ppm of copper should meet the copper requirement of sheep and cattle, respectively, unless amounts of molybdenum and sulfur intake are high. A copper:molybdenum ratio of 2.0 or greater is desirable to avoid molybdenosis (Miltmore and Mason, 1971; Ward, 1978).

A deficiency of copper in cattle occurs when the

dietary level is much less than 5 ppm DM or when molybdenum or sulfur are in excess (Ward, 1978). Ward (1978) reported that dietary copper:molybdenum ratios less than 2:1 contribute to a copper deficiency, whereas an excess of dietary sulfur potentiates the effect of molybdenum.

Cobalt is the metal cofactor in vitamin B12 which is in turn required in energy metabolism in ruminants (Mayland et al., 1987). Pasture herbage levels of at least 0.08 and

0.11 ppm will provide adequate cobalt for cattle and sheep, respectively (Grace, 1983). Ruminants on Phalaris pastures can develop an acute form of a disease from which they quickly die or a chronic form of a nervous disorders characterized by muscle tremors, rapid breathing and pounding heart beat. Phalaris staggers is the name given to this disorder. Grazing Phalaris increases the cobalt requirement (Mayland et al., 1987).

The toxicity and metabolism of molybdenum are dependent not only on the levels of dietary molybdenum but also on the levels of other dietary components. The higher the level







of molybdenum, the greater the amount of copper required to prevent signs of molybdenosis. Prolonged high molybdenum intakes cause hypocupremia (Underwood, 1981).

Molybdenum-induced copper deficiency is an endemic problem in ruminants (Ward, 1978). Dietary levels of molybdenum are affected by parent soil material, soil pH, forage type and forage maturity (Reid and Horvath, 1980). Molybdenum combines with hydrogen sulfide in the rumen to form thiomolybdates which render copper unavailable for absorption (Dick et al., 1975).

In many areas sheep and cattle grow normally on pastures containing 0.03 ppm of selenium and show no evidence of a deficiency. In other areas, white muscle disease can occur, especially in lambs, where pastures contain as much as 0.05 ppm. These responses may be attributed to variations in dietary sulfur levels or other factors that affect the absorption of selenium or the requirement of selenium by the animal. The requirement for selenium in sheep is given by the National Research Council (NRC, 1985) as 0.1-0.2 ppm. A value of 0.1 is often used as a critical level, but this should be evaluated on a basis of animal performance (Mayland et al., 1987). Selenium deficiency is associated with pastures that contain less than 0.03 ppm (Millar, 1983).









Mineral Status and Requirements of Ruminants


Table 3 presents the requirements and critical levels for the diagnosis of specific mineral deficiencies or toxicities in sheep. Without question, forage analysis is a much better indicator of mineral status for ruminants than is soil analysis. Likewise, animal tissue-mineral concentrations are better indicators of the availability of minerals than are forage mineral analyses. Grazing livestock obtain part of their mineral supply from the consumption of water, soil, leaves, tree bark, etc, rather than entirely from forages (McDowell et al., 1986).

Evaluating the mineral status of domestic animals can be a complex and costly procedure. Often little is known about the nutritional background of animals in question. Collecting blood or tissue samples for mineral analysis is often impractical because of limited facilities and personnel trained to collect samples (Combs, 1987). Tissue Macrominerals

Calcium is the most abundant of the minerals in the animal body. About 26-30% of total ash content of most animals is calcium. Although about 98-99% of the total body calcium is in the skeleton, it has numerous crucial functions in soft tissues. The active form of calcium in soft tissues is the ionized form. Ionized calcium content of blood plasma is homeostatically regulated within












TABLE 3. DIAGNOSIS OF SPECIFIC
TOXICITIES IN SHEEP



Dietary requirement


Element


Deficienc

Calcium



Phosphorus



Magnesium Potassium Sodium Sulfur Iodine Iron



Copper


Molybdenum Cobalt Manganese Selenium


(dry basis)o


.20-.82 %



.16-.38 %



.12-.18 % .50-.80 % .09-.18 % .14-.26 % .10-.80 % 30-50 ppm



7-11 ppm .5 ppm .1-.2 ppm 20-40 ppm .1-.2 ppm


MINERAL DEFICIENCIES OR


Tissue


Bone (fat free) Bone ash Plasma Bone (fat free) Bone ash Plasma Serum Urine



Saliva



Milk Hemoglobin Transferrin


Liver Serum



Liver Liver Liver Serum Hair or wool


Critical levelb~c


24.5 % 37.6 %
8 mg/dl

11.5 % 17.6 %
4.5 mg/dl

1-2 mg/dl
2-10 mg/dl



100-200 mg/dl



300 ug/day

10 g/dl 13-15 % saturation

25-75 ppm .65 ug/ml



.05-.07 ppm

6 ppm

.25 ppm .03 ug/ml .25 ppm











TABLE 3.--CONTINUED


Dietary requirement
Element (dry basis)8 Tissue Critical levelb'c Toxicity

Copper 25 ppm Liver 700 ppm Fluorine 60-150 ppm Bone 4500-5500 ppm Manganese 1000 ppm Hair 70 ppm Iron 500 ppm

Molybdenum 10 ppm Liver 4 ppm Selenium >2 ppm Liver 5-15 ppm Hair 10 ppm

Zinc 750 ppm Hair 10 ppm ONRC (1985). Requirements below which a deficiency occurs. bReferences for critical levels are found in the following reviews: Mtimuni (1982); McDowell et al. (1984). cNon-mineral assays for the following elements are sensitive diagnostic techniques: cobalt (Vitamin B1), iodine (free thyroxine), copper (ceruloplasmin) and selenium (glutathione peroxidase).







relatively narrow limits. The skeletal system serves as a very effective reservoir of calcium, which maintains ionized plasma calcium levels within narrow limits under a wide range of dietary calcium intakes (Combs, 1987).

Calcium, phosphorus and magnesium are important

components of bone, and also intracellular and extracellular fluids of the body. Extracellular calcium is essential for maintenance of nerve tissue, resting membrane potential, blood clotting mechanisms, myocardial contraction and myoneural junctional transmission. Intracellular calcium, directly or indirectly, regulates activity of many enzymes, microtubule assembly, generation of ATP, release of hormones and neurotransmitters and muscle cell contraction (Littledike and Goff, 1987). Excess calcium may impair reproductive function by causing a secondary deficiency of phosphorus, magnesium, zinc, copper and other microelements by inhibiting their absorption in the intestine (King, 1971).

Phosphorus has long been recognized as a major

essential nutrient for ruminants. Approximately 80% of body phosphorus occurs in bone and skeletal development depends upon an adequate supply of phosphorus. Phosphorus is required for phosphorylation in sugar metabolism, intracellular energy transfer, formation of phospholipids, as a buffer in blood and other fluids and is required for proper functioning of rumen microorganisms (Cohen, 1987).







Phosphorus is also abundant in animal tissues, accounting for 16-17% of total body ash.

Severe deficiencies of phosphorus in cattle reduce feed intakes, feed efficiencies, and retard growth (Kincaid et al., 1981). The requirement for phosphorus as a percent of dry matter in rations for sheep is given by the National Research Council (NRC, 1985) as 0.16-0.38%.

Dietary calcium:phosphorus ratios may affect

reproductive performance. Phosphorus deficiency induces lowered conception rate, irregular estrus, decreased ovarian activity, increased incidence of cystic follicles, and generally depressed fertility. When phosphorus levels are low, phosphorus supplementation, while expensive, is critical to animal performance (Hurley and Doane, 1989).

There has been increasing acceptance of the rib bone biopsy technique (Little, 1972) as a more reliable method for the estimation of phosphorus status of grazing cattle and sheep than blood. McDowell (1985) has indicated that this technique is being used widely as a survey technique to locate mineral deficiencies in tropical regions.

About 70% of total body magnesium in livestock occurs in the bones. It also occurs in high concentrations in intracellular fluid where it is associated with the mitochondria and in lesser concentrations in the extracellular fluid. It is involved in oxidative phosphorylation, phosphate transfer, metabolism of carbohydrates and lipids and in neuromuscular activity







(Cohen, 1987). Magnesium accounts for 1-1.1% of total ash of most animal species and is deposited primarily in skeletal and muscle tissue. Seven to eight % of the total body stores of magnesium is in other tissues and body fluids (Combs, 1987).

Magnesium is an essential cofactor of many enzymes, especially phosphate-transferring enzymes involved in ATP generation and the adenylate and guanate cyclases, and is essential for normal function of nerve tissues (Littledike and Goff, 1987).

The magnesium status of livestock can be assesed from plasma, whole blood or urine (Egan, 1980). Normal magnesium levels in plasma of sheep are listed as 1.8-2.0 mg/dl with values below 1.0 mg being severely hypomagnesemic (NRC, 1985). The National Research Council (1985) suggested minimum magnesium requirements of 0.12, 0.15 and 0.18% of dry matter for growing lambs, for ewes in late pregnancy, and for ewes in early lactation, respectively. Studies have shown that excess magnesium intake by ruminants caused loss in weight, drowsiness and changes in blood mineral levels (Gentry et al., 1978).

Hypomagnesemic tetany is a problem in ruminants managed under a variety of regimens. The disorder has been reported in milk-fed calves, and in adult sheep and cattle fed highroughage diets or maintained on sparse pasture (Giduck and Fontenot, 1987). A deficiency of magnesium in ruminants may result from low magnesium concentrations in feeds or a







reduction in biological availability of dietary magnesium (Fontenot, 1982).

Sodium, potassium and chlorine function in maintaining acid-base balance, osmotic pressure and body fluid balance. Sodium is involved in transmission of nervous impulses and occurs largely in the body fluids and bones while potassium occurs mainly in the muscle, nervous tissue and erythrocytes (Cohen, 1987).

Sodium deficiency leads to a pica or craving for salt, unthrifty appearance, loss of appetite, weight loss and reduced milk yield. These signs usually occur without a decline in plasma sodium, although urinary and fecal sodium may decline (Underwood, 1981). The National Academy of Sciences (NRC, 1985) listed sodium requirements at 0.090.18% dry matter of the diet and potassium at 0.50-0.80% dry matter.

Tissue Microminerals

Zinc occurs widely and in relatively high

concentrations throughout the body. Zinc concentrations in plasma of sheep and cattle range from 0.6 to 1.2 ppm, respectively. Zinc is a constituent of a large number of metallo-enzymes involved in biochemical processes essential to nucleic acid and carbohydrate metabolism, as well as protein synthesis. It is associated with appetite, growth, male sexual development and wound healing. There are no significant stores of body zinc, and the animal must rely on a daily supply to meet requirements (Mayland et al., 1987).





30

Largely on the basis of Australian studies, Underwood (1981) concluded that zinc requirements for optimum growth and fertility of sheep must lie close to 30 ppm of the dry diet. The NRC (1985) requirements of sheep are given as 29 to 33 ppm.

Clinical signs of severe zinc deficiency have been

reported in ruminants under practical conditions. However, a marginal zinc deficiency appears to be a more widespread occurrence (Spears, 1989). Low zinc intake throughout pregnancy has severe effects on reproduction in the ewe. Because signs of zinc deficiency are nonspecific, poor zinc status should be considered in cases of unexplained reproductive problems (Apgar and Fitzgerald, 1985). Excess dietary iron can affect performance adversely in ruminants. High dietary iron can affect utilization of other minerals such as copper, phosphorus, zinc and manganese (Humphries et al., 1983).

The liver is the primary storage organ of body copper stores, having about 40-70% of the total copper. Copper is active in the conversion of tyrosine to melanin, which provides the color pigment in hair and wool. About 20% of the plasma copper is in a loosely bound form, -while the other 80% is associated with a protein called ferroxidase I (Ceruloplasmin). Ferroxidase I oxidizes ferrous iron (Fe2 ) to ferric (Fe3 ) allowing the mobilization of iron stores (Mayland et al., 1987).







Adequate maternal intake of copper is essential for development of the central nervous system of the embryonic lamb. Enzootic ataxia of the unborn or the unweaned lamb is primarily from copper deficiency (Hidiroglou and Knipfel, 1981). Visible signs of copper deficiency are not usually seen in the adult sheep. Copper deficient lambs may have a degenerative disorder known as swayback (Poole, 1982).

Physiological copper deficiencies are produced by four classes of feeds: (1) high molybdenum, generally above 100 ppm, (2) low copper:molybdenum ratio, 2:1 or less, (3) copper deficiency, below 5 ppm, and (4) high protein, 2930% protein in fresh forage (Ward, 1978).

Copper toxicity is essentially a problem of the housed ruminant because housed ruminants are given foodstuffs of high copper availability. Conversely, copper deficiency is a problem of the grazing animal because of the poor availability of copper in grass, so low as to be less than 10% of that found in some foodstuffs (Suttle, 1986).

Sulfur in the inorganic form, which is converted within the gastrointestinal tract to certain amino acids and Bvitamins by rumen microorganisms, can meet the needs of ruminants for sulfur. This process provides methionine, thiamin and biotin, which otherwise would need to be provided by the diet to meet the needs of sulfur-containing compounds for the animal's metabolic, regulatory and structural functions. All other sulfur-containing compounds







required by mammalian tissue can be synthesized from methionine (Goodrich and Thompson, 1981).

Elevated molybdenum intakes depress copper availability and may produce a physiological copper deficiency in ruminants. Total sulfur or sulfate in the ration generally potentiates the effect of molybdenum. The ratio of copper to molybdenum in feed is important regardless of the absolute amount of each. For this reason, and because of the importance of the sulfur content of the diet, it is impossible to define safe dietary limits of copper and molybdenum (Ward, 1978).

Thiomolybdates reduce the absorption of dietary copper in sheep. They also affect systemic copper metabolism by changing markedly the distribution of copper in plasma and by reducing the availability of copper to metabolic sites within the body (Gooneratne et al., 1989).

According to Mayland et al. (1987) loss of appetite,

high aspartic aminotransferase, and low blood plasma glucose levels are the best indicators of cobalt deficiency. Serum vitamin B12 less than 0.20 ng/ml will also indicate a possible cobalt deficiency, but liver cobalt levels are not always reliable estimates of cobalt status.

Selenium is widely distributed in the body. The kidney and liver normally have the highest concentrations. Selenium is an integral part of the enzyme glutathione peroxidase which catalyzes the reduction of peroxides, thereby protecting tissues against oxidative damage.







A dietary concentration of 0.1-0.2 ppm of selenium has been accepted by the NRC (1985) as a safe and adequate level for the prevention of white muscle disease in sheep. Workers in New Zealand found that lambs grew normally with no signs of white muscle disease when pastures contained

0.03 to 0.04 ppm of selenium (Hartley and Grant, 1961). However, Whanger et al. (1978) reported white muscle disease in lambs fed diets containing 0.1 ppm of selenium.

High levels of oral copper have been reported to reduce selenium availability in nonruminants. Copper often is added to mineral mixes for ruminants, and copper toxicosis is relatively common in sheep (Underwood, 1981). White et al. (1989) showed that copper or molybdenum supplements at 10 mg/kg to practical-type diets of ewes and lambs had no effect on selenium status.



Hematological Measurements



Blood is the principal fluid transport system in the

body and provides an expedient source of metabolic products. Chemical analysis of serum and plasma provide quantitative estimates of physiological parameters for diagnosis of disease. Quantitative differences among normal cows have been found for many of these variables by Peterson et al. (1982).

Hemoglobin is the oxygen-carrying compound contained in red blood cells (RBC). The amount of hemoglobin per 100 ml







of blood can be used as an index of the oxygen-carrying capacity of the blood. Total blood depends primarily on the number of RBCs (the hemoglobin carriers) but also, to a much lesser extent, on the amount of hemoglobin in each RBC (Ravel, 1989). Reference values are most frequently quoted in sheep as 9-15 g/dl (Table 4).

Since whole blood is made up essentially of RBC and plasma, the percentage of packed RBCs after centrifugation gives an indirect estimate of the number of RBCs/100 ml of whole blood. Hematocrit thus depends mostly on the number of RBCs, but there is some effect (to a much lesser extent) from the average size of the RBC (Ravel, 1989).

White blood cells (WBC or leucocytes) form the first

line of defense of the body against invading microorganisms. Neutrophils and monocytes respond to phagocitosis; lymphocites and plasma cells apparently produce antibodies (Ravel, 1989).








TABLE 4. NORMAL BLOOD HEMOGLOBIN, HEMATOCRIT AND LEUCOCYTE
VALUES FOR THE SHEEP


Range'


Average


Hemoglobin, g/dl 9-15 11.5 Hematocrit, % 27-45 35 Leucocytes, /ul 4000-12,000 8000

Neutrophil (mature), % 10-50 30 Lymphocyte, % 40-75 62

Monocyte, % 0- 6 2.5

Eosinophil, % 0-10 5

Basophil, % 0- 3 0.5 aSchalm and Nemi (1986).















CHAPTER III
MATERIALS AND METHODS



Identification and Description of Research



The research for the present study was conducted at three sheep farms located in the paramo region of the Cordillera Oriental of Colombia during both the rainy and dry seasons.

San Jorge farm belongs to Instituto Colombiano

Agropecuario (ICA) and is located in Soacha (Cundinamarca), on one dry area of the paramo. Don Benito and San Francisco farms belong to Caja de Credito Agrario Industrial y Minero (Caja Agraria) and are located in Zipaquira (Cundinamarca) and Ventaquemada (Boyaca), respectively, on wet areas of the paramo (Appendix B, table 20). About 36% of the total sheep population, or 85% of the wooled sheep of the Country are located in these two departments (Cundinamarca and Boyaca). Location of the farms is shown in Appendix A, figure 2.

Soil, forage and animal tissue samples were collected from the three farms during the rainy and dry seasons. The two seasons were selected based on the pattern of rainfall. There are two relatively short but marked rainy seasons during the year: from mid-April to mid-June and then from







mid-September to mid-November. Appendix B table 20 shows the climate and average monthly rainfall of two of the farms. The first sampling period corresponded to the end of the rainy season (May, June 1987) and the second sampling corresponded to the middle-end of the dry season (February 1988).

Identification and a general description of parameters in evaluating animal production for each farm appears in appendix B table 19. This information was provided by farm managers during the sample collection period in 1988. Some data were derived from an approximation by the manager in cases where exact information was not available.


Sample Collection



Soil Samples

A total of 113 composite samples were collected from the three farms during both 1987 and 1988 sampling periods. Each composite sample was made up of 8-12 samples taken from predetermined areas of a paddock, totalling 4-5 composite samples from each paddock.

Twenty, nineteen and twenty composite samples from

different paddocks were collected from San Jorge, Don Benito and San Francisco, respectively, during 1987 (rainy season) sampling period. Twenty, fourteen and twenty composite







samples from different paddocks were collected from San Jorge, Don Benito and San Francisco, respectively, during 1988 (dry season) sampling period.

Samples of soil were taken from the top layer (20 cm) using a stainless steel bore. A soil sampling technique described by Bahia (1978) was used. Although soil samples collected during the two seasons did not come from the exact same spot, they came from the same grazing area of the farm.

Based on texture, the soil of the upper part (above

3,000 m) of San Jorge is classified as loamy (about 50%) or silt loam (about 50%); in the lower part (below 3,000 m), about 40% of the soil is clay, 40% is clay loam and sandy clay loam, and the rest is sandy clay, loam, and sandy loam. In Don Benito the predominant soil is loamy, and in San Francisco it is either loamy or silt loam.

Proper identification of each paddock is in appendix C, and the detailed number of composite soil samples is in appendix B table 21.

Soil and forage samples were collected one or two days before animal tissue samples were taken. Approximately 500 g composite soil were transferred to plastic bags and were properly identified for further analysis at the Soils Laboratory at Tibaitata, ICA, Colombia. Twenty gram subsamples were taken, transferred to small plastic bags and were properly identified for selenium analysis in the United States.







Forage Samples

A total of 131 composite forage samples were taken from the three farms during both the rainy season (1987) and the dry season (1988). Twenty-eight, twenty and nineteen composite samples containing the major species of forage from San Jorge, Don Benito and San Francisco, respectively, were taken during the 1987 (rainy) collection period. Twenty-nine, fifteen and twenty composites from San Jorge, Don Benito and San Francisco were taken during the 1988 (dry) collection period. The forage species collected were: vernalgrass (Anthoxanthum odoratum), a native cultivar of velvetgrass (Holcus lanatus L), an imported cultivar of velvetgrass (H. lanatus basyn), kikuyugrass (Pennisetum clandestinum), white clover (Trifolium re _), tall fescue (Festuca arundinacea), and orchardgrass (Dactvlis glomerata).

In both collections forage samples were taken from the same areas where soil samples were taken. Appendix B table 21 shows the detailed number of forage samples; proper identification of paddocks and of forage species is in appendix C.

Each of the farms maintains about 3 sheep/ha/year under a rotational grazing system (appendix B table 19). There is not a fertilization program in San Jorge; however, some paddocks in the lower part of the farm receive nitrogen (in the form of Urea) which is applied at 50 kg ha"1. In Don Benito and San Francisco pastures are not fertilized;







however, some paddocks take advantage of residual fertilization as a consequence of potato cropping.

Each of the composite forage samples from each paddock came from 20-25 individual samples of the same forage species predominating and most frequently grazed by sheep on the different areas of the farm.

To avoid contamination, plastic gloves were utilized for forage collection. Only the aerial part of the forage (about 5 cm from the ground) was taken. Samples were collected in plastic bags and kept refrigerated at 50 C until further processing. Samples were then hand separated by species and within each species samples were further separated into two parts, stems and leaves. A third component of the sample was left totally unseparated and analyzed separately as "whole plant", but no comparisons were made with the stem and leaf components of the same plant. Plant parts were transferred to paper bags and oven dried at 600 C for 48 hours. After this process, the samples were ground in a hammer mill with stainless steel knives and 1 mm screen at ICA facilities in Bogota. After mixing, a 60 g sample was transferred into a plastic bag and properly identified for further chemical analysis in the Nutrition Laboratory of the University of Florida. Animal Tissue Samples

Blood serum, whole blood, bone and liver samples were collected from sheep on each farm. The animals were divided







into three classes: lactating (or pregnant) ewes, lambs (14 months of age) and yearlings (10-14 months of age).

Because of timing differences in the breeding seasons, it was not possible to collect samples from lambs in San Francisco at the 1987 collection period because they were very young or not yet born; in this case their mothers were in their last trimester of pregnancy. Appendix B table 23 details the number of animal tissue (serum, whole blood, liver, and bone) samples taken.

The animals in San Jorge were Criollo x Romney or Criollo x Corriedale crosses and in Don Benito and San Francisco the animals were Criollo x Blackface crosses (appendix B table 19).

During each sampling period, animals from the desired classes were selected at random from each farm. Because it was not possible to follow the animals sampled during the rainy season of 1987, other animals were randomly selected in the dry season of 1988. Animal stress and excitement were minimized during and prior to tissue sample collection.

The composition of the mineral mixture given on the

three farms is presented in appendix B table 22. A majority of the animals received mineral supplementation although this was not done for the entire year especially in San Jorge farm.

Whole blood and blood serum. Duplicate blood samples were obtained by jugular puncture and were collected in monovette tubes (Sarstedt, West Germany). One of them







contained the anticoagulant NH4-heparin. This tube was inverted 5 times so the blood made contact with the anticoagulant but avoiding hemolysis of the red cells. The blood samples were left standing in a cool environment and sent to Laboratorio de Investigaciones Medico Veterinarias (LIMV) in Bogota (ICA, Colombia) for the determination of hematocrit, hemoglobin and total and differential leucocyte counts.

The second tube was used for serum separation. The

blood samples were centrifuged at 2500 rpm for 30 minutes at the respective farm. The serum samples were identified and kept frozen after centrifugation until the precipitation of serum proteins was completed at the University of Florida. Procedures and techniques for blood processing have been described by Fick et al. (1979). In total, 207 serum and 192 whole blood samples were analyzed.

Bone bioosY A total of 148 bone biopsy samples were taken from the sheep. Bone biopsy samples were taken as described by Little (1972), with some modifications of the technique. A single sample was removed from the 12th rib on the right side of the animal. The procedure performed using the same vertical incision, approximately 4 cm in length made for the liver biopsy, was used for the bone biopsy. The width of the rib in sheep is so narrow that the biopsy was taken without using a trephine; instead, a piece of bone approximately 2 cm in length was removed by cutting a section of rib using a small stainless steel bone shear.







Bone samples were kept in individual plastic bottles containing 10% formalin for analysis at the University of Florida.

Liver biopsy. A total of 113 liver biopsy samples

were taken from the sheep. Samples were taken in vivo using the technique described by Fick et al. (1979) and McDowell et al. (1983).

The same animals on each farm selected for blood

sampling were used for liver biopsy sampling. Liver samples were kept frozen in 10% formalin for further analysis at the University of Florida.



Chemical Analysis


Soil Samples

Soil samples were analyzed by standard methods in the Laboratorio Nacional de Suelos in Tibaitata, ICA, Bogota for organic matter (OM), pH, aluminum, boron, calcium, copper, iron, magnesium, manganese, phosphorus, potassium, sodium, and zinc.

Aluminum (exchangeable acidity) was extracted from the soil samples with the solvent KCI, IN; exchangeable basis were extracted with ammonium acetate and organic matter was determined by the Walkley and Black colorimetric method (Black, 1967). Phosphorus was measured using the method of Bray II (Jackson, 1958).






Soil mineral concentrations were determined by atomic absorption spectrophotometry (Perkin-Elmer, 1980). Selenium was analyzed at the University of Florida, by a fluorometric method (Whetter and Ullrey, 1978). Foraae Samples

Forage samples were processed and analyzed for mineral concentration according to methods described by Fick et al. (1979) at the Nutrition Laboratory, University of Florida. Calcium, copper, iron, magnesium, manganese, potassium, sodium and zinc were analyzed by atomic absorption spectrophotometry in a Perkin-Elmer AAS 5000 (Perkin-Elmer, 1980). Cobalt and molybdenum were analyzed by flameless atomic absorption spectrophotometry using a Perkin-Elmer AAS Zeeman/3030 (Perkin-Elmer, 1984). Phosphorus was determined by the colorimetric method described by Harris and Popat (1954) and included by Fick et al. (1979) as a method for phosphorus determination for plant and animal tissues. Selenium in forage samples was analyzed by a fluorometric method (Whetter and Ullrey, 1978).

For nitrogen analysi.s, samples were digested using a modification of the aluminum block digestion procedure of Gallaher et al. (1975). Sample weight was 0.3 g, catalyst used was 3.2 g of 9:1 K2SO4 : CuSO4, and digestion was conducted for 4 h at 4000 C using 10 ml H2S04 and 2 ml H202. Ammonia in the digestate was determined by semiautomated colorimetry (Hambleton, 1977). Multiplication of nitrogen by 6.25 was the procedure for calculating crude protein. In







vitro organic matter digestion (IVOMD) was performed by a modification of the two-stage technique (Moore and Mott, 1974). Dry matter was determined by drying for 15 h at 1050 C and organic matter by ashing for 15 h at 5500 C. Animal Tissue Samples

Blood serum. Serum brought to the University of Florida was deproteinized with 10% trichloroacetic acid (TCA) and then analyzed for mineral content according to the method described by Fick et al. (1979). Calcium, copper, magnesium, and zinc were analyzed by atomic absorption spectrophotometry using a Perkin-Elmer AAS 5000 (PerkinElmer, 1980). Phosphorus was determined by a colorimetric technique (Harris and Popat, 1954). Selenium was analyzed using the method of Whetter and Ullrey (1978).

Whole blood. Blood samples which contained

anticoagulant were used to determine hematocrit, hemoglobin, and total and differential leucocyte counts in Laboratorio de Investigaciones Medico Veterinarias (LIMV), ICA, Colombia. The microhematocrit method was utilized using capillary tubes of 1.1-1.2 mm diameter in an International centrifuge with standardized reading scales. Centrifugation was for 5 minutes at 2500 rpm. After centrifugation, the height of the RBC column was measured and compared with the height of the column of original whole blood. The percentage of RBC mass to original blood volume is the hematocrit (Ravel, 1989).







The hemoglobin concentration was obtained utilizing the oxyhemoglobin method. A .025 ml aliquot of blood is added to a test tube containing 5 ml sodium bicarbonate (.1% solution) and mixed (Schalm and Nemi, 1986). A Leitz spectrophotometer was utilized.

To determine total leucocyte counts, a Coulter Counter model FN (Coulter Electronics, Inc, Hialeah, Florida) was used. The Coulter Counter is based on the principle that cells are poor electrical conductors. A measured volume of diluted suspension of cells (in an electrically conductive medium) is drawn through a minute aperture between two electrodes. Each cell passing through the aperture displaces an equal volume of the electrolyte solution and is counted electronically and displayed on the digital readout (Schalm and Nemi, 1986). Differential leucocyte counts were appraised visually from prepared slides of blood smears using the methods described by Schalm and Nemi (1986).

Bone samples. The samples were dried and extracted in ether following procedures outlined by Fick et al. (1979) and subsequently analyzedfor calcium, magnesium, and phosphorus.

Liver samples. Livr biopsy sample preparation was carried out as described by Fick et al. (1979). Dry tissue samples (approximately .3 g) were pre-ashed on a hot plate with 50% (v/v) nitric acid and then ashed overnight in a muffle furnace at increments of 1000 C every hour until reaching 5500 C. Ash was solubilized first with 50% nitric





47

acid, then with 10% nitric acid and finally, with distilled water. Solutions were filtered, diluted to appropiate range and analyzed with atomic absorption spectrophotometry for copper, iron, manganese, and zinc using a Perkin- Elmer AAS 5000 (Perkin-Elmer, 1980). Liver cobalt and molybdenum were determined by flameless atomic absorption spectrophotometry using a Perkin-Elmer AAS Zeeman/3030 (Perkin-Elmer, 1984). Selenium analysis was carried out by the technique of Whetter and Ullrey (1978).



Statistical Analysis



Data were analyzed by use of the Statistical Analysis System (SAS Institute., 1985). Probability level for significance was .05 in all statistical analyses. Blood Serum. Whole Blood. Bone and Liver

Serum, whole blood, bone and liver were analyzed as a split-plot design with animal class as the main plot and season as the subplot. The model was as follows:

Yijk = u + Ai + BJ + Cij + Dk + Fki + Gkj + Ejk , where: u = overall mean.

Ai = random effect of the ith farm. Bi = effect of the jth animal class. Cii = effect of farm*animal class (experimental error for

animal class).

Dk = effect of the kth season. Fki = effect of season*farm.







Gki = effect of season*animal class. Eljk = residual error (experimental error associated with

the subplot).

Since the data were unbalanced, hypothesis testing was based on the Type IV Sum of Squares. Soil and Foraae

Soil and forage were analyzed as a split-plot design, with farm as the main plot and season as the subplot. For forage, plant part was considered as the subplot. The model was as follows:

Yijk = u + Aj + Bj0) + Ck + Dik + Eijk , where: u = overall mean.

Ai = fixed effect of ith farm. B = effect of jth paddock within ith farm (experimental

error for farm).

Ck = effect of kth season. (kth plant part, in forages). Di= effect of farm*season. (farm*plant part, in forages).

Eijk = experimental error associated with subplots.

Correlation coefficients between soil and forage, forage and animal tissues, and serum, liver and bone responses were estimated. These estimates were obtained for each farm and class separately for the rainy and dry seasons.

Only those correlation coefficients of biological importance are discussed in the corresponding chapters.













CHAPTER IV
MINERAL STATUS OF SHEEP IN THE PARAMO REGION
OF COLOMBIA. I. MACROELEMENTS


Introduction



Sheep are adapted to the cold and harsh environment of the Colombian paramo; but it is commonly accepted that the few forage species that are grown are of low quality and their regrowth is very slow after grazing. Under these conditions animals do not get their nutritional requirements and, therefore, the sheep enterprise is not very efficient. Poor growth rate of lambs, low fertility in particular of imported breeds, high mortality and low wool production of inferior quality are characteristics of sheep production in the Colombian paramo (Proyecto Ovino Colombo Britanico, 1979).

Mineral deficiencies, imbalances, and toxicities have been reported to severely inhibit tropical cattle production systems (McDowell, 1985). It has been demonstrated that, with the exception of calcium and sulfur, none of the macroelements have adequate concentrations in some forage species grazed by sheep on the paramo (Laredo et al., 1989). The objectives of this study were to evaluate the macromineral status of grazing sheep and to determine macroelement status and other soil, forage, and blood







parameters in three farms as related to the wet and dry seasons of the Colombian paramo.



Materials and Methods



Soil, forage, and animal tissue samples were collected from three sheep farms in the paramo of the Cordillera Oriental of Colombia. Sampling periods corresponded to the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988).

A total of 113 composite soil, 131 composite forage, 207 serum, 192 whole blood, and 148 rib bone biopsy samples were obtained for each of the sampling periods.

Each composite soil sample came from 8-12 samples. A soil sampling technique described by Bahia (1978) was used. Soil samples were analyzed by standard methods in the Laboratorio Nacional de Suelos of ICA in Bogota for organic matter, pH, aluminum, calcium, magnesium, phosphorus, potassium and sodium. Minerals were extracted from the soil samples with an acid extracting solution (.025N H2SO4 and .05N HCl).

Based on texture, the soil of the upper part (above

3,000 m) of San Jorge is classified as loamy (about 50%) or silt loam (about 50%); about 40% of the soil of the lower part (below 3,000 m) is clay, 40% between clay loam and sandy clay loam; the rest is sandy clay, loam, and sandy







loam. In Don Benito the predominant soil is loamy, and in San Francisco it is either loamy or silt loam.

Each composite forage sample came from 20-25 samples of the same forage species predominating and most frequently grazed by sheep in the different areas of the farms. Forage species collected were: vernalgrass (A. odoratum), native velvetgrass (H. l L), imported velvetgrass (H. lanatus basyn), kikuyugrass (P. clandestinum), white clover (T. repens), tall fescue (E. arundinacea), and orchardgrass (_. alomerata). Each of the farms maintains about 3 sheep/ha/year under a rotational grazing system (appendix B table 19). There is not a fertilization program in San Jorge; however, some paddocks of the lower part receive nitrogen (in the form of Urea) at 50 kg ha'1 year. In Don Benito and San Francisco pastures are not fertilized; however, some paddocks take advantage of residual fertilization as a consequence of potato cropping. Forage samples were processed and analyzed for mineral content according to methods described by Fick et al. (1979).

Samples were collected from animals that were divided into three classes: lactating (or pregnant) ewes, lambs (14 months of age) and yearlings (10-14 months). The sheep were Criollo x Romney or Criollo x Corriedale in San Jorge and Criollo x Blackface in Don Benito and San Francisco.

Duplicate blood samples were obtained by jugular

puncture and were collected in monovette tubes (Sarstedt,






West Germany). One tube contained the anticoagulant NH4heparin. The second tube was used for serum separation.

Bone biopsy samples were taken as described by Little (1972), with some modifications of the technique. The biopsy was taken without using a trephine; instead, a piece of bone approximately 2 cm in length was removed by cutting a section of rib using a small stainless steel bone shear.

Forage calcium, magnesium, potassium and sodium, serum calcium, magnesium, copper and zinc and bone calcium and magnesium were determined by atomic absorption spectrophotometry using a Perkin-Elmer AAS 5000 (PerkinElmer, 1980). Forage, serum and bone phosphorus were determined by the colorimetric method of Harris and Popat (1954). For nitrogen analysis, forge samples were digested using a modification of the aluminum block digestion procedure of Gallaher et al. (1975). In vitro organic digestibility (IVOMD) was performed by a modification of the two-stage technique (Moore and Mott, 1974).

Whole blood samples were processed at the Laboratorio de Investigaciones Medico Veterinarias (LIMV) of ICA in Bogota for hematocrit, hemoglobin and total and differential leucocyte counts. Hemoglobin concentration was obtained utilizing the oxyhemoglobin method, and to determine total leucocyte counts, a Coulter Counter model FN was utilized (Schalm and Nemi, 1986).

Data were analyzed by use of the Statistical Analysis System (SAS Institute, 1985). Soil and forage were analyzed







as a split-plot design, with farm as the main plot and season as the subplot. For forage, plant part was considered the subplot. Animal tissues were analyzed as a split-plot design with animal class as the main plot and season as the subplot. Significance level was limited to .05 in all statistical analysis. Since the data were unbalanced, hypothesis testing was based on the Type IV Sum of Squares. Correlation coefficients among minerals were determined.

Results and Discussion


Soil Analyses
Soil analyses as related to season and farm are

presented in table 5. Summary of the analysis of variance is shown in appendix B table 24. Soil organic matter was lower (P<.05) in San Jorge farm during the rainy season, but the pH was higher (P<.05) for the same farm during the dry season. Reid and Horvath (1980) indicated that maximum rate of cation absorption occurs at pH 5-7. Soil acidity problems are associated with pH levels lower than 5.5 and the presence of exchangeable aluminum in the soil (Sanchez, 1976). Don Benito had a higher (P<.05) aluminum concentration during the rainy season; this farm had the lowest soil pH (4.8) of the three farms.

Among the macroelements, soils were most deficient in magnesium in relation to critical levels. Percentage of samples below the critical level of 30 ppm suggested by Rhue


















SuoITOGIuGIuo1 101(1143 Jo UOOITOWIGoIdJI oIsopIIII o AlrUOpon busaq.,ae osoIl 1inAoO - .abd .B J0 Sjlos i01 GTrlco1dd0 0q wou A Pue p81o01 40 sijos .pu.s .1oj ..0 l-o pAA-pap GIG. - all001 1-,10113 sOGZ
� (W0"Od) lO]JTP dason ua Tp 11! -d I ~ IO .. uTj ~ .-sIs a q sueoh,6 ( q ( s o ' O > d ) .0 TJ s.pd f . . . . ..s 1 - q.l a jjp p q 4 . .r -n 1 oS U S u I S U G AI lo q s u a Jr h



"(9L61) puOIGa
.u.1 o... p0p-04Sq
�1osTouOf ups 0o (Alp 'AuTe) O OZ P-0 0T1(08 uo00 l04 (Alp 'AaT(0) 81 '61 '-1 GaboO uS -J (Alp A'TI) IL O :d olosodmoo o -aOZ Tl.o IloJ GI u4 paoq 8ue08


VTB 11 "0 8?" 91"0 S"Z E*E 0"0 3 860 PC ,
19 8Lg OL 90"0 ts 001 UT E 1 Lf 9 LL 15 L19 E9 0 [L'8 LO 00 9L C'89 001 SO'I 08? E6 ZI VII O 19 8'6L 06 06,1 LL 89 6WE izsz 0z 61 8"SZ SE GS'E 9"8Z Iz 18 t "VlI 00 9v 8"6 s0 OF11 0'61 ZI S0 9"1S 00 LT 11S 01 88 16z 09 1Z[ (?Z 0 O0 4188 01 188 81? "E 199 lzsZ 0

89[ 0"1[ 8z8 CI8 618 to[ S'51 91E L'LL aGL5 1"0 10"0 6,0 80"0 8a 8 0 s C0"0 0"s so 'O 88 V81 E6"0 11? "LIO 9"61 9"61 60 L'LZ VLI iO


111 I/LI Alp 09"1 a8*ZE Au T[11 801 A p 6"01 S9 UT-a pZ9 s 6z 10 Alp 9"Z? 1[6I A.!-1 01 L6"Z L[l AIp
6 681 Ap pL


011 096 Alp Iot 10o6 Aur, IL

99" 1 6 Alp


LO'O 00"5 Ap 00,0 1 0s oo 0[I/ 0"z1 Alp LLI J90o AlOar


[ o w j NS ]- u.3. OG T s u -W 'Jac % G2op sw u .. -aS a
1 .. lA-S l~ua uo 1-.4il


"(SISq IHA) osva
uNv NOSVaS Oi uaivaa SV SHSAqVNV qVU9NIWOHDVW GNV 'H(I 'UHIVN DIMVDHO qIOS "S aqUVI


."d Ix .dd 56W odd ',j 8dd I4O .dd *IV


Hd


% "11I OW a106eA

111(01101







and Kidder (1983), was 61 for the rainy season and 76 for the dry season. There were differences among farms with San Jorge having the highest levels (P<.05). Metson (1974) stated that there is some evidence that acidity aggravates a magnesium deficiency condition.

In general, San Jorge had higher concentrations (P<.05) during the rainy season for calcium, magnesium, and sodium. On the other hand, San Jorge had the lowest phosphorus level but it was not significant (P>.05). This element was 46% deficient in the rainy season and 39% in the dry season. Many tropical soils are generally reported to be deficient in phosphorus (Volkweiss, 1978).

Thirty seven % of the samples in the rainy season and 61% in the dry season were below the critical level of 62 ppm suggested for potassium by Rhue and Kidder (1983). Sodium presented very low levels in the three farms especially in Don Benito and San Francisco. Forace Analyses

Forage macromineral, crude protein and IVOMD

concentrations are presented in table 6. Summary of the analysis of variance is presented in appendix B table 25. Means and standard errors of each species in a season, are presented in appendix B tables 35 and 36. Even though the forage samples were separated by species, these could not be included in the statistical model because of the uneven occurrence of some of them which produced a large number of empty cells.















TABLE 6. FORAGE MACROMINERAL, CRUDE PROTEIN AND IVOMD CONCENTRATIONS AS RELATED TO
SEASON AND FARM (DRY BASIS).


Variable


Critical San Jorge Don Benito San Francisco Overall level Season Mean S.E.b Def. Mean S.E. % Def. Mean S.E. t Def. Mean % Del.


C.. % 0.2c rainy 0.50 0.07 4 0.31 0.04 5 0.24 0.02 37 0.37 13 dry 0.35 0.05 13 0.25 0.02 40 0.29 0.02 20 0.31 22 K, B v 0.5' rainy 2.75 0.18 0 3.28 0.14 0 2 19 0.12 0 2.75g 0 dry 1.53 0.11 3 1.64 0.19 0 1.03 0.06 0 1.40h 1 Mg, % v 0.12' rainy 0.21 0.01 21 0.11f 000 40 0.13 0 00 37 0.16q 31 dry 0.13' 0.1 50 0.00 0.00 100 0.09' 0.00 95 0.11h 75 Na. % v 0.09' rainy 0.04 0.00 89 0.03 0.00 100 0.05 0.00 89 0.03 93 dry 0.04 0.00 97 0.05 0.01 87 0.03 0.00 95 0.04 94 F, % < 0.16' rainy 0.28 0.01 7 0.30 0.01 0 0.20 0.00 0 0,27q 3 dry 0.17 0.01 47 0.16 0.01 60 0.12 0.00 85 0.15h 62 CP, % 7d rainy 17.5 1.00 0 20.6 0.97 0 17.4 0.89 5 18.49 q dry 11.7 0.79 13 14.7 1.82 7 10.6 0.62 10 12.0h 11 1VOM, 8 rainy 58.7' 2.03 71.1 1.00 72.7' 1.22 66.7 dry 57.6 1.36 58.9 2.58 53.8 1.10 56.7

aMeans based on the following number of composite samples: 28, 29 (rainy, dry) for San Jorge, 20, 15 (rainy, dry) for Don Benito and 19, 20 (rainy, dry) for San Francisco.
bStandard error of neans.
CNRC (1985).
".--o ad Milford (1)7 .

-"M M- ee s eto, e farm, - i iow, n with dfifff r, v,,t t rrfilrys differ d(10.05). ,) hMens bet-ee -....n ill a -o- with different scup ...ripts differ (<)o)







Lower limits of sheep requirements (dry matter basis) suggested by the NRC (1985) were used to calculate the percentage of deficient forage samples. Sodium was the most deficient macroelement in forage samples (93-94%) according to the critical level of .09% (NRC, 1985). Potassium was the least deficient (only 1%) element in forage samples. This element is often associated with high levels of nitrogen and moisture in lush, cool-season grasses (Boling et al., 1979). Magnesium was 31 and 75% deficient for rainy and dry season, respectively. Phosphorus was only 3% deficient in the rainy season but 62% in the dry season. Underwood (1981) stated that phosphorus deficiency is the most widespread and economically important of all the mineral deficiencies affecting grazing livestock. Thirteen and 22% of forage samples were calcium deficient in the rainy and dry seasons, respectively . The results of this study are in contrast to the results of many researchers in the Latin American tropics who have reported relatively low calcium and extremely deficient phosphorus forage concentrations (McDowell,. 1985).

Surprisingly, only 1% of the samples in the rainy

season and 11% in the dry season were deficient for crude protein, according to the critical value of 7% suggested by Minson and Milford (1967). The high protein concentration found in this study agrees with that of Laredo and Anzola (1986) who determined the crude protein level in temperate grasses similar to the ones used in this research. Sanchez







(1976) stated that organic matter in the soil supplies most of the nitrogen and half of the phosphorus taken up by unfertilized crops; Sillanpaa (1982) stated that the correlation between the nitrogen content of the plant and total soil nitrogen is relatively good.

Magnesium in San Jorge forage samples was higher

(P<.05) during the dry season but the deficiency for this farm was as high as 50%. Michael (1962) reported that serum magnesium levels in sheep were not correlated with herbage magnesium content, but Fontenot (1982) stated that a magnesium deficiency in ruminants may result from low magnesium concentrations in feeds or a reduction in biological availability of dietary magnesium. Nitrogen level, stage of maturity, excessive potassium level, form of magnesium, and amount of readily fermentable carbohydrate have been considered as components of magnesium utilization (Rosero et al., 1980).

Calcium, potassium, phosphorus, sodium and crude

protein concentrations were not different (P>.05) among the three farms. The IVOMDwas lower (P<.05) for San Jorge during the rainy season but not during the dry season. The values of IVOMD for the forages in San Jorge were not low (58.7 and 57.6) and those for Don Benito (71.1 and 58.9) and San Francisco (72.6 and 53.8) are considered high, which implies that the animals were receiving good quality forage especially during the rainy season. Since IVOMD values were relatively high it seems that production of forage DM/ha in







the paramo is more of a limitation to animal performance than is forage quality. In a previous study, Laredo et al. (1989) found that the production of DM/ha in Don Benito was 2.8 ton and 2.0 ton for H. lanatus basyn (imported cultivar) and H. lanatus L (native cultivar); in San Francisco the production of DM/ha was 2.8 ton and 0.8 ton, respectively. These values were low for the standards of the grass (Watt, 1978) and even lower when compared to other temperate grasses.

There was a marked effect of season on all

macroelements studied. Magnesium, potassium, phosphorus and crude protein were lower (P<.05) during the dry season. Although sodium was not different (P>.05) between seasons, concentrations indicated that there was a serious deficiency at all times. Calcium had similar values for both seasons. McDowell et al. (1983) stated that as plants mature, mineral concentration declines due to a natural dilution process and translocation of nutrients to the root system; in most circumstances, magnesium, phosphorus, potassium and sodium decline as the plant matures. Crude protein declined from 18.4% to 12% (P<.05). In vitro organic matter digestibility also declined, from 66.7% to 56.7%, but the diference was not significant (P>.05).

Animal Tissue Analyses

Blood serum and bone macromineral concentrations as related to season and farm and as related to season and







animal class are presented in tables 7 and 8. Results of analysis of variance are presented in appendix B table 26.

Blood serum. No differences were found (P>.05) among farms for serum calcium and phosphorus during both the rainy and dry seasons, and for magnesium during the rainy season. Magnesium had a higher value (P<.05) in farm San Jorge (2.18 mg/dl) than farm San Francisco (1.93 mg/dl) during the dry season.

Differences (P<.05) among animal classes were found for magnesium and phosphorus. Lambs had higher (P<.05) concentrations for serum phosphorus than ewes (5.2, 5.6 vs 3.6, 2.5 mg/dl) in the rainy and dry seasons, respectively, and higher (P<.05) than yearlings but only in the dry season (4.0 mg/dl). On the contrary, lambs had lower (P<.05) values for serum magnesium than yearlings in the dry season. There were no differences (P>.05) for serum calcium among the three animal classes.

The general tendency was for the dry season to have lower values of calcium, magnesium and phosphorus; nevertheless, phosphorus was the only element statistically affected (P<.05) by season (4.71 vs 3.99 mg/dl).

The incidence of calcium deficiency as percentage of

samples below critical levels (McDowell et al., 1984) was 3% for the rainy season and 94% for the dry season. The incidence of deficiency for San Jorge, Don Benito and San Francisco during the dry season was 100, 97 and 87%, respectively; the incidence of deficiency for ewes, lambs















TABLE 7. BLOOD SERUM AND BONE MACROMWNERAL CONCENTRATIONS AS RELATED TO SEASON AND FARM.





San Jore lDon Benito _ San Francisco Overall Variable I-elr Season Meanb S.. Def. Mean S.E Def. Mean S.E. % Def. Mean % Def.


8 rainy 10.09 0.12
dry 6.20 0.10 4 C5 rainy ; 4.72 0.23
dry 3.87 0.22
2 rainy 2.21 0.04
dry 2.18d 0.06
reel

66.8 rainy 62.1 0.51
dry 59.4 1.62
24.5 rainy 22.06 0.29
dry 20.15 0.56
11.5 rainy 7.81 0.27
dry 5.92 0.28 rainy 0.37 0.01 dry 0,32 0.00


3 9.95 0.18
100 6.08 0.13

53 4.96 0.29 71 4.41 0.23

8 2.05 0.07 41 1.98 0'05


6 10.34 0 12 97 6.49 0.18

42 4.37 0.37 62 3.76 0 25

33 2.17 0.05 47 1.93e 0.05


0 10.1 3 87 6.28 94

58 4 71 51 67 3.99 67

25 2.14 22 61 2.02 51


100 62.3 0.55 00 63.4 0.42 100 62.6 100 85 65.2 1.12 88 62.4 0.38 96 61.7 89

100 22.19 0.29 100 21.99 0.15 100 22.1 100 100 23.34 1.26 8 21.08 0.28 100 21.2 97

100 6.76 0.67 95 7.26 0.31 100 7.32 99 100 8.20 0.55 94 6.91 0.26 100 6.73 99
0.38 0.02 0.35 0.01 0.36 0.43 0.03 0.39 0.01 0.37


MDowel et a. (19841.
bMeans based on the foll-aing number of samples: ...r 36, 34 (rainy, dry) for San Jore farm, 33, 34 (rainy, dry) fo- fln "onito and 24, 46 (rainy, dry) for San Franisco. Bone 26, 33 (rainy, dry) for San Jorqe; 22, 16 (rainy, dry) for Dot Benito and 25, 28 (rainy, dry) for San Francis-o.
VStandard error of ears.
d'emeans a gos9 farms in a row with different supersrripts differ (1r0.051 f"'Means between seasons in a rolnen with different sutvrscriPts differ (-rO.05)


Serim C., q/dl P. mq/dl


Mg, -q/dO Bone (1.1., fat-f
Ash, %


P, %













TABLE 8. BLOOD SERUM AND BONE MACROMINERAL CONCENTRATIONS AS RELATED TO SEASON
AND ANIMAL CLASS.




CrVaitl 6 Em. es LD Ms Yearlings Overal
Vaibelevela Season Meanb C .c EDef. Mean 5.12 % DcC. Meav S EC % DeC. Mean r ef.


Ca, e/dl P, sg/dl Mg, nq/dl

Cone (D.M., fat-free
Ash, I

Ca. I C, 8 89, %


8 rainy 9.69 0.15 a 10.13 0.16 0 0 53 0.09 0 10.1 3 dry 5.95 0.14 100 7.14 0.24 74 6.05 0.04 100 6.28 94 4.5 rainy 1.62e 0.19 84 5.18d 0.30 38 5.58d 0.24 23 4.71f 51 dry 2.54 0.16 97 5.62" 0.26 19 4.00' 0.12 72 3.99g 67 2 rain5 2.15 0 05 27 2.01 0.09 29 2.21d 0.04 11 2.14 22 dry 1.91, 0.07 52 1.84 0.04 81 2.14 0.04 35 2.02 51


66.s rainy 64.2d 0.32 100 60.3f 0.60 100 61.7' 0.37 100 62.6 100
dry 65.8 0.40 70 59.4 1.51 100 63.6 0.51 95 61,7 89 24.5 rainy 22.8 0.18 100 21.3 0.19 100 21.5 0.23 100 22.1 100
dry 22 A 0.20 100 16.7 0.54 100 21.9 0.52 95 21.2 97 11.5 rainy 7.84 0.22 100 5.25 0.58 100 7.60 0.47 96 7.32 99
dry 7.32 0.16 100 4.77 0.28 1o0 7.02 0.32 98 6.73 99
rainy 0.34 0.01 0.40 0.01 0.37 0.01 0.36 dry 0,14 0.01 0.32 0.01 0.40 0.01 0.37


aMcDowell et al. (1984).
"Mean based on the following number of sample u..sern 37, 33 (rainy, dry) for ewes, 21, 27 (rainy, dry) for lamb, and V5, 54 (rainy, dry) for yearlings; Bone 34, 20 (rainy, dry) for ewes, 14, 13 (rainy, dry) for laus and 25, 44 (rainy, dry) for yearlings. CStandard error of mans.
Means aemog aniMa classes in a row with different superscripts differ (P,0.05). ghrMeans between seasons in a olun with different superscripts differ (Pr0.05).







and yearlings was 100, 74, and 100%, respectively during the dry season. The high incidence of serum samples below the critical level indicated that the sheep had a very severe calcium deficiency during the dry season. Sheep and cattle have hormonal mechanisms which maintain blood calcium concentrations within narrow limits by adjusting the proportion of dietary calcium absorbed and, when dietary calcium is inadequate, by resorbing calcium from body reserves in the skeleton (Rowlands, 1980). Black et al. (1973) reported that serum calcium concentrations may be directly affected by dietary calcium intake, and Steevens et al. (1971) reported that serum calcium concentrations are affected more by the amounts of phosphorus and magnesium in the diet than by calcium itself. Serum calcium, however, is influenced only by severe deficiency, and calcium dietary levels may be a more adequate criterion in assessing status of calcium (CMN, 1973).

Serum inorganic phosphate was deficient in 51% of

samples during the rainy season and of 67% during the dry season according to the level of 4.5 mg/dl suggested as a critical concentration by McDowell et al. (1984). Deficiency of phosphorus for farms San Jorge, Don Benito, and San Francisco was 53, 42 and 58% for the rainy season and 71, 62, and 67% for the dry season, respectively. Deficiency of serum phosphorus for ewes, lambs and yearlings was 84, 38 and 23% for the rainy season and 97, 19 and 72% for the dry season. Plasma inorganic phosphate







concentrations are maintained by absorption of phosphorus from the gut, and there is no specific mechanism for bone phosphorus resorption (Jacobson et al., 1972); positive relationships between dietary phosphorus intake and plasma inorganic phosphate concentrations have been observed (Rowlands, 1980). However, serum or plasma phosphate is not recommended as a practical criterion for assessing phosphorus status in cattle or sheep (CMN, 1973).

Overall incidence of samples below the critical level of 2 mg/dl of magnesium suggested by McDowell et al. (1984) was 22% for the rainy season and 51% for the dry season. The deficiency for farms San Jorge, Don Benito and San Francisco was 41, 47 and 61% for the dry season. For ewes, lambs and yearlings the deficiency was 52, 81 and 35% for the dry season. In contrast to calcium, serum magnesium concentration depends mainly on the dietary intake of available magnesium (Rowlands, 1980). Levels below 2 mg/dl in plasma are classed as hypomagnesaemic, but magnesium concentration in blood serum does not fall until there is a severe deficiency (CMN, 1973).

Bone. Bone macromineral concentrations as related to season and farm and as related to season and animal class are presented in tables 7 and 8. Summary of analysis of variance is presented in appendix B table 26.

There was no season nor farm effect (P>.05) for any of the bone parameters studied with the exception that ash







during the rainy season had a variation among animal classes. Ewes had higher (P<.05) ash concentrations than yearlings (64.2 vs 61.7%), and lambs lower (P<.05) than both classes at 60.3%.

Individual evaluation of samples indicated that ash

concentrations below a critical level of 66.8% (McDowell et al., 1984), was 100% during the rainy season and 89% during the dry season. Spongy bone of the axial skeleton are the first to demineralize in periods of negative balance (Little, 1972).

Calcium mean concentrations were below the suggested critical level of 24.5% (McDowell et al., 1984). Deficiencies were 100% for the rainy season and 97% for the dry season. This is in agreement with the results observed by other researchers (Knebush et al., 1986, Tejada et al., 1987) who reported that when animals were evaluated for calcium status, using blood serum and bone samples, a consistently higher number of animals were determined to be below the critical levels based on bone analysis when compared to critical levels in serum. Furthermore, this supports the concept that cattle or sheep resist depletion of plasma calcium through mobilization of bone calcium, thus making bone a more accurate indicator of calcium levels in the animal.

Mean rib bone phosphorus was 99% deficient in both rainy and dry seasons, according to the critical level of 11.5% (McDowell et al., 1984). This is in agreement with







the results of other researchers who worked with cattle in the tropics (Tejada et al., 1987, Vargas et al., 1984). Adequate forage phosphorus concentration appears to be reflected more accurately in plasma phosphate than bone, even though many researchers prefer bone phosphorus over serum concentrations to evaluate the phosphorus status of an animal (McDowell et al., 1983).

Bone magnesium concentrations were similar among farms and among animal classes. Cohen (1987) suggested that bone magnesium may be useful in assessing magnesium status in grazing livestock.

Correlation Coefficients of Minerals

Blood serum and bone macromineral correlation

coefficients as related to season are presented in appendix B table 28. Correlation coefficients (P<.05, r>1.501) in serum were calcium-phosphorus (r=.76), calcium-iron (r=.65) and calcium-zinc (r=.77) during the dry season. For bone, the correlations were calcium-ash (r=.79) during the rainy season, calcium-phosphorus (r=.80), and calcium-ash (r=.97) during the dry season..

Correlation coefficients between serum and bone

minerals as related to season are presented in appendix B table 29. Correlation coefficients between serum and bone macrominerals were observed among serum phosphorus-bone magnesium (r=.84) and serum magnesium-bone phosphorus (r-.85) during the rainy season.







In general, mineral concentrations among animal tissues did not correlate with each other with a correlation coefficient greater or equal to 1.501 at the probability level of .05. This demonstrated the problem of finding significant correlation coefficients between soil, forage and animal tissues (Conrad et al., 1980). Hematological Measurements

Hemoglobin and hematocrit concentrations and leucocyte counts as related to season and farm are presented in table 9. Summary of analysis of variance is in appendix B table 27.

San Jorge had higher (P<.05) total leucocyte counts (16,838/ul) than Don Benito (7,605/ul) and San Francisco (8,107/ul) during the rainy season. During the dry season the values were much lower for San Jorge and different only to San Francisco.

Total leucocyte counts in San Jorge during the rainy season exceeded the normal limit of range in sheep set by Schalm and Nemi (1986) of 12,000/ul. Differential leucocyte count in the same farm, during the rainy season, showed that the percentage of lymphocytes exceeded the limit of range of 75% (Schalm and Nemi, 1986).

There were no differences (P>.05) among animal classes for any of the hematological measurements studied, but lambs in the rainy season exceeded the limit of range for total leucocytes (Schalm and Nemi, 1986) and also exceeded the limit of range of 75% for lymphocytes but were below the




















TABLE 9. HEMOGLOBIN, HEMATOCRIT COINCER-TTION AND LEUCOCYTE COUNTS AS RELATED TO SEASON AND FARM.




gS Jore Don benit an Francisco Overall nange no
Variable levelsa Season Meanb S.E.' Mean S.E. Mean S. Mean

Hematocrit, % 27-45 rainy 34.0 ).51 40.8 0.67 40.3 0.66 38.0 dry 34.6 0.64 37.6 0.74 39.5 0.69 37.4
Hemoglobin, q dl 9-15 rainy 11.4 .5 13.6 22 13.4 0.22 12.6 dry 11.6 0.32 12.7 0.21 13.4 0.30 12.6 Leooocynes. j 4000-12000 rainy 16838d 997 1605e 322 8107: 285 11458 dry 7277d 316 6671 369 4598 186 6089 Neutrohils, 1 10-50 rain '.8 1. 16.6 1.7 6, 1.97 18 0 or'/ ..4 .4 221 1.3 71.1 4 2. Lympnocytes, * 40-75 rainy 84.8 1.43 76.5 2.04 65.0 1.9e 77.1 dry 71.5 2.68 73.1 2.23 70.3 2.67 71.5 Monocytes, % 0-6 rainy 0.72 0.15 1.50 0.21 2.00 0.66 1.3 dry 1.11 0.22 1.34 0.24 0.63 0.17 1.0 Eosinophils, a 0-10 rainy 1.72 0.48 3.28 0.79 7.00 0.97 3.5 dry 2.56 0.50 2.66 0.55 2.88 0.53 2.7 Basophils, a 0-3 rainy 0.00 - 0.09 0.06 0.00 - 0.,e dry 0.42 0.11 0,76 0.17 0.23 0.08 0.44d aschalm and Nems (19861

bMeans based on the following number of samples: 36, 36 (rainy, dry) for San Jorge, '2. 29 (rainy, dry) for Don Benito and 21, 40 (rainy, dry for San Francisco. cStandard error of meansd''eMeans anong trms n a row with different super ropts differ P005) .







range of 10% for mature neutrophils percentage. These lambs exhibited slight to moderate leucocytosis, defined as WBC values of 13,000 and 20,000/ul (Holman, 1944) and could mean that an infectious process was taking place in the lambs of San Jorge at the time the samples were taken.



Summary and Conclusions



A study was conducted to determine the macromineral

status of three sheep farms located on the paramo region of the Cordillera Oriental in Colombia, and to compare animal classes (pregnant-lactating ewes, lambs, and yearlings). Soil, forage, blood and rib bone samples were collected to correspond to the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988).

Season affected (P<.05) soil concentrations of calcium and sodium and forage concentrations of magnesium, potassium, phosphorus and crude protein. Soil analyses showed high organic matter (19%) and low pH values (5.0) for the three farms. Magnesium was the most deficient macroelement in both seasons with an overall deficiency of 70%, followed by potassium with 52% of the samples deficient; phosphorus was 46% deficient. Sodium was the most deficient macrolement in forage samples with 93% overall deficiency; phosphorus was deficient only in the dry season (62%) and magnesium had 53% overall deficiency. Crude protein was deficient in 6% of the forage samples.







Blood serum analyses showed an overall phosphorus

deficiency in 59% of the samples, and a calcium dry season deficiency of 94%. Bone was 98% deficient in both calcium and phosphorus, with ash percentage being also very deficient (95%). Differences (P<.05) among animal classes were found in serum phosphorus in both seasons (lambs were higher in phosphorus), in serum magnesium in the dry season (lambs were lower in magnesium), and bone ash in the rainy season (lambs were lower in ash percentage).

Among soil minerals and the corresponding forage

minerals only calcium and magnesium had positive correlation (P<.05, r > 1.501) coefficients for both seasons. In general, macrominerals between animal tissues did not correlate with each other.

Based on these analyses it was concluded that

macromineral status of sheep on the paramo was deficient and supplementation programs should provide common salt, calcium, phosphorus and magnesium.

















CHAPTER V
MINERAL STATUS OF SHEEP IN THE PARAMO REGION
OF COLOMBIA. II. MICROELEMENTS


Introduction



It has been reported (McDowell and Conrad, 1977) that ruminants grazing forages in severely mineral-deficient areas may even be more limited by this condition than either a lack of energy or protein, and that trace element deficiencies or imbalances in soils and forages are responsible for low production and reproduction among grazing livestock (McDowell et al., 1984). As grazing livestock usually do not receive mineral supplementation except for common salt, they must depend almost exclusively upon forages for their requirements. Only rarely, however, can forages completely satisfy all mineral requirements (Miles and McDowell, 1983).

It has been shown from a region in the Colombian paramo that forages are low in copper, cobalt and zinc and high in iron and molybdenum and that this imbalance in micronutrients might be the cause for the low production of the sheep in the area (Proyecto Ovino Colombo Britanico, 1979).







The objectives of this study were to evaluate the

micromineral status of grazing sheep and to determine the microelement status of soil and forages of three sheep farms as related to the two seasons prevailing in the paramo region of Colombia.



Materials and Methods


Soil, forage and animal tissue samples were collected from three sheep farms in the Cordillera Oriental of the paramo of Colombia. Sampling periods corresponded to the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988).

A total of 113 composite soil, 131 composite forage, 207 serum and 113 liver biopsy samples were obtained for each of the sampling periods.

Each composite soil sample for each farm was obtained from 8-12 samples. A sampling technique described by Bahia (1978) was used. Soil samples were analyzed by standard methods in the Laboratorip Nacional de Suelos of ICA in Bogota for boron, copper, iron, manganese and zinc. Minerals were extracted from the soil samples.with a double acid extracting solution (.025N H2SO4 and .05 N HCl). Soil mineral concentrations were determined by atomic absorption spectrophotometry (Perkin-Elmer, 1980).

Based on texture, the soil in the upper part (above

3,000 m) of San Jorge is classified as loamy (about 50%) or







silt loam (about 50%); in the lower part (below 3,000 m) about 40% of the soil is clay, 40% a combination of clay loam and sandy clay loam, and the rest is sandy clay, loam, and sandy loam. In Don Benito the predominant soil is loamy, and in San Francisco is either loamy or silt loam. Each composite forage sample was obtained from 20-25 samples of the same forage species predominating and most frequently grazed by sheep in the different areas of the farm. The forage species collected were vernalgrass (A. odoratum), a native cultivar of velvetgrass (H. lanatus L), an imported cultivar of velvetgrass (H. lanatus basyn), kikuyugrass (P. clandestinum), white clover (T. rjj), tall fescue (F. arundinacea), and orchardgrass (D. glomerata). Appendix B table 21 shows the detailed number of soil and forage species; proper identification of paddocks and of forage species is in appendix C. Forage samples were processed and analyzed for mineral content according to methods described by Fick et al. (1979).

Each of the farms maintains about 3 sheep/ha/year under a rotational grazing system. There is not a fertilization program in San Jorge; however, some paddocks in the lower part of the farm receive nitrogen (in the form of Urea) at 50 kg/ha/year. In Don Benito and San Francisco pastures are not fertilized; however, some paddocks take advantage of residual fertilization as a consequence of potato cropping.

Samples were collected from animals that were divided into three classes: lactating (or pregnant) ewes, lambs (I-







4 months of age) and yearlings (10-14 months of age). The sheep were Criollo x Romney or Criollo x Corriedale in San Jorge and Criollo x Blackface in Don Benito and San Francisco.

Blood plasma and liver biopsy samples were collected from the animals at each farm as described by Fick et al. (1979) and McDowell et al. (1983).

Selenium analyses of soil, forages, blood and liver

were carried out using a fluorometric technique described by Whetter and Ullrey (1978).

Forage iron, copper, manganese and zinc, blood copper and zinc, and liver copper, iron, manganese and zinc were determined using atomic absorption spectrophotometry using a Perkin-Elmer AAS 5000 (Perkin-Elmer, 1980). An atomic absorption spectrophotometer equipped with a graphite furnace and Zeeman background corrector (Perkin-Elmer Model 3030) was used to determine forage and liver cobalt and molybdenum.

Data were analyzed by use of the Statistical Analysis System (SAS Institute, 1985). Soil and forage were analyzed as a split-plot design, with farm as the main plot and season as the subplot. For forage, plant part was considered as the subplot. Animal tissues were analyzed as a split-plot design with animal class as the main plot and season as the subplot. Significance level was limited to .05 in all statistical analyses. Since the data were unbalanced, hypothesis testing was based on the Type IV Sum







of Squares. Correlation coefficients between minerals were estimated.



Results and Discussion



Soil Analyses

Soil micromineral analyses as related to season and farm are presented in table 10. Summary of analysis of variance is shown in appendix B table 30. Description of paddocks and forage species are in appendix C.

Soil iron was higher (P<.05) in Don Benito than San

Francisco during the dry season. A similar trend was found during the rainy season. According to the critical level of

2.5 ppm set by Viets and Lindsay (1973) none of the samples for the three farms were deficient in iron. These concentrations were considerably in excess of normal values, and could result in a reduced availability of phosphorus to plants (Lindsay, 1972).

San Francisco farm had a lower (P<.05) soil zinc value than the other two farms during the rainy season. Individual evaluation of samples based on a critical level of 1 ppm when pH is below 6.5 (Rhue and Kidder, 1983) indicated that none of the samples during the rainy season, and only 13% during the dry season were deficient for this element.

San Francisco had a lower (P<.05) soil boron

concentration than the other two farms during the dry















TABLE 10. SOIL MICROMINERAL ANALYSES AS RELATED TO SEASON AND FARM (DRY BASIS).


CritcalSan Jore On Benito level Se.n.. Mean, S.E b I Def. Mean S.E. % Def.


0 660 77.9
0 622 135

0 2.6 0.70 0 0.4 0.08 30 6.2 0.89 35 4.5 0.49 0 2.8' 0.16 15 1.8 0.2) 100 0.05 85 0.28 0.02 60 o.3o 0:02


San Francisco


Overall


Mean S.E. % Def. Mean % Def.


32.7 0 19.5 0 0.12 0 0.12 5

0.62 50 0.32 70 0.11 0 0.04 5
100
0.03 80


0.28 85


79 0.219 0.02 10 0.28 80


aMeans based on the following number of composite samples: 20, 20 (rainy, dry) for San Jorqe farm, 19, 14 (rainy. dry) for on Benito and 20, 20 (rainy, dry) for San Francisco.
b
Standard error of mean. Cviets and Lindsay (1973). dRhos and Kidder (1983). eGamman (1976).

f'qMeans among farns in a row with different superscripts differ (PiO.05). hMcoowel et al. (1989).

*Selenium was not included in the statistical analyses. i1Means between seasons in a column with different supcrscripts differ (Pi0.05).

These critical levels were derived for the more sandy soils of Florida and may not be appropriate for soils of the parano. one.er, these
are being used only as guides or approsimations of critical concentrations.


rainy dry


Variable Fe. ppn Cu, ppm


Mn, ppn Zn, pp. Se , pp. B, pp.


453 387f9


0.3 5d id



.5h

0.4


40.5 13.3
0.35 0.26

2.11
1.42

0.22
0.27


rainy 2.0 dry 2.0 rainy 12.5 dry 10.5 rainy 2.7f dry 2.2 rainy 0.03 rainy 0.28 dry 0.35







season. The percent deficiency of this element, according to the value of 0.4 ppm suggested by Gammon (1976) was 85% for the rainy season and 80% for the dry season. Boron has been established as essential for higher plants and is added frequently to fertilizers for plants with high requirements such as alfalfa (NRC, 1980).

Farm was not a source of variation (P>.05) for copper and manganese during both seasons, for boron and iron during the rainy season and for zinc during the dry season. The only microelement affected (P<.05) by season was zinc which had a lower value during the dry season. Copper, iron and manganese had lower values during the dry season but were not different (P>.05).

The percentage of samples below the critical level of .3 ppm for copper (Rhue and Kidder, 1983) was none during the the rainy season and 17% during the dry season. Most of the deficient samples for copper were found in Don Benito, with 57% of the samples deficient during the dry season.

For manganese, the percentage of samples below the critical level of 5 ppm (Rhue and Kidder, 1983) was 44% during the rainy season and 59% during the dry season. Among farms the deficiency was San Jorge, 30 and 35; Don Benito 53 and 79; San Francisco, 50 and 70% for rainy and dry seasons respectively.

Soil selenium was not included in the statistical

analysis but the concentrations for the three farms during the rainy season are presented in table 10. The selenium







concentration of soil reflects that of the parent material (Reid and Horvath, 1980). Some research has indicated that soil selenium concentrations of less than .5 ppm are found in areas where selenium deficiency in livestock occurs (McDowell et al., 1989). Based on this critical level, all samples were deficient in selenium. Forage Analyses

Results of forage micromineral analyses as related to farm and season are presented in table 11. Summary of analysis of variance is shown in appendix B table 31.

Forage cobalt was higher (P<.05) in San Jorge in

comparison to Don Benito and San Francisco during the rainy season. A similar tendency was observed during the dry season, but was not significant (P>.05). According to the critical level of .1 ppm suggested by the NRC (1985), 79% of the samples were deficient in cobalt during the rainy season and 95% during the dry season. Among farms deficiencies were San Jorge, 54 and 90; Don Benito, 100 and 100; San Francisco 95 and 100% for rainy and dry seasons, respectively. McDowell et al. (1982) reported that, with the exception of phosphorus and copper, cobalt deficiency is the most severe mineral limitation to grazing livestock in many tropical countries, and sheep are more severely affected than other ruminant species.

Manganese concentration was higher (P<.05) in San

Francisco in comparison to San Jorge during the dry season. According to the value of 20 ppm (NRC, 1985) only 1% of the



















TABLE 11. FORAGE MICROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND FARM (DRY
BASIS).




Critical San Jorge Don Benito San Francisco Overall Variable level Season Meana S.E.b Def. Mean S.. % Def. Mean S.E. % Def. Mean % IOf.


C pp 11 1 7c rainy 7.52 0.65 36 6.91 0.43 65 6.97 064 79 7.18 57 dry 2.92 0.34 97 3,87 0.52 87 2.70 0.36 95 3.07 94 Co. pipm 0.1 rainy 011 0.01 54 0.01f 0.00 100 S 04f 0.00 95 0.06 79 dry 0.06 0.02 90 0.03 0.00 100 0.03 0.00 100 0.05 95 Fe, pa 30c rainy 113 16.2 14 106 14.5 0 89 9.2 0 104 6 dry 119 11.7 0 144 18.1 0 150 12.2 0 135 0 M., pp. 20c rainy 167 16.0 4 228 17.1 0 231 26.4 0 203 1 dry 211 17.6 0 381e 31.4 0 402e 27.9 0 309 0 MB, ip[ I 6d rainy 1.87f 0.14 - I.59' 0.18 - 0.66' 0.06 1.03 dry 0.44 0.09 - 0.79 0.18 - 0.64 0.11 0.58 Se, pam 0.1 rainy 0.11' 0.01 50 0.15e 0.01 25 0.09f 0.01 53 0.11 43 dry 0.10 0.01 70 0.10 0.01 53 0.08 0.01 80 0.o9 69 Zn. pin 20' rainy 244 2.24 11 24.3 1.11 20 19.4 0.87 63 25.0 28 dry 18.3 1.32 57 26.2 2A10 13 20.4 2.05 50 20.7 45

aMeans based on the fc1l.owinq numbr of sample- 28, 29 (rainy, dry) for San Jorqe, 20, 15 (rainy, dry) f-r Don Benito and 19, 20 (raiy, dry) for San Francisco.
"Standard error of means
CNc (1985).
d
McrOsalI et al. (1984).
e'flh'Bans amnqg farms it, a ro with diffcin, t . i.rsciits differ F1P-O.P ). "'hMan- betwen seasons in a con sith different soynrscripts differ (P,0.05).





80

samples were deficient during the rainy season and none during the dry season. All values were in excess of the requirements for sheep. Several studies indicate the ruminant has a high tolerance for manganese (Hansard, 1983); mineral imbalances typified by excesses of iron and manganese may interfere with metabolism of other minerals (Lebdosoekojo et al., 1980). The farm means were well below the maximum tolerable level of 1000 ppm (NRC, 1985).

Forage selenium was higher (P<.05) in Don Benito

compared with the other two farms during the rainy season. Forty-three % of the samples during the rainy season and 69% during the dry season were deficient according to the .1 ppm value suggested for sheep by NRC (1985). Among farms the deficiencies were: San Jorge, 50 and 70; Don Benito, 25 and 53; San Francisco, 53 and 80% for rainy and dry seasons, respectively. Underwood (1981) considered that for all ordinary grasses and legumes the primary determinant of selenium concentration was the level of available selenium in the soil.

Forage molybdenum concentrations were different (P<.05) among farms during the rainy season. Don Benito had a higher value in comparison with the other two farms. However, none of these values were above the 6 ppm (dry basis) suggested as a toxic limit (McDowell et al., 1984). According to Suttle (1986) only a small increase in molybdenum and sulfur concentrations will cause major reduction in copper availability. He reported that







differences of 3 ppm of molybdenum (from 1 to 4 ppm) and .5 ppm of sulfur (from 2.5 to 3) between two pastures are sufficient to reduce copper availability to one half.

Farm was not a source of variation (P>.05) for copper, iron and zinc during both seasons; for cobalt, molybdenum and selenium during the dry season and for manganese during the rainy season. Season was a source of variation (P<.05) for the following microelements: cobalt, copper, manganese and molybdenum; all except manganese had lower (P<.05) concentrations during the dry season. The levels of selenium and zinc were also lower during the dry season, but were not significant (P>.05). On the contrary, iron and manganese were higher during the dry season.

The percentage of samples below the critical level of 7 ppm suggested by NRC (1985) for copper was 57% for the rainy season and 94% for the dry season. Among farms the deficiencies were as follows: San Jorge, 36 and 97; Don Benito, 65 and 87; San Francisco, 79 and 95% for rainy and dry seasons, respectively. The results of this study showed that copper:molybdenum ratio was at least 6:1 in San Jorge and San Francisco and 4:1 in Don Benito, and that molybdenum levels were not higher than 4 ppm in any case. Copper concentration was marginal in forages during the rainy season and deficient during the dry season. The animals, in this situation, might respond to appropiate copper supplementation.







Evaluation of samples based on the dietary zinc

requirement of 20 ppm (NRC, 1985) indicated deficiencies of 28% during the rainy season and of 45% during the dry season. Among farms, the deficiencies were: San Jorge, 11 and 57; Don Benito, 20 and 13; San Francisco, 63 and 50% for rainy and dry seasons. Ruminants have exhibited signs of zinc deficiency when grazing forage containing 20 to 30 ppm of zinc (Pierson, 1966). However, a marginal zinc deficiency appears to be a more widespread occurrence (Spears, 1989).

Forage samples evaluated on the levels of 30 ppm of iron (NRC, 1985) showed that only 6% were deficient during the rainy season and none during the dry season. This is in agreement with the zero incidence of iron deficiency found in the soil samples. McDowell (1985) stated that ruminant animals are not likely to suffer from iron deficiency. None of the samples reached the maximum tolerable level of 500 ppm suggested for sheep by the NRC (1985). Animal Tissue Analyses

Blood serum and liver micromineral concentrations as

related to season and farm are presented in table 12, and as related to season and animal class are in table 13. Summary of analysis of variance of serum and liver microminerals is shown in appendix B table 32.

Blood serum. There were no significant farm by animal class interactions (P>.05) for any of the serum microminerals considering seasons separately. Iron,












TABLE 12. BLOOD SERUM AND LIVER MICROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND FARM.




San _Jore Don Benito San Franciso overall Variable levela Seaso Me..b S.C.' % Def. Mean S.E. %rf. Mea. S.E. Dof. Mea. Def.



Cu. ppm o 0.6 rainy 0.79 0.04 19 0.69 0.04 10 1.04 0.03 0 0.82 18 dry 1.01 0.03 0 1.20 0.05 0 1,34 0.05 0 1.20 0 Zn, pm < 0.6 rainy 0.70 0.03 22 0.75 0.03 12 0.85 0.04 4 0.76 14 dry 0.78 0.10 24 0.68 0.03 32 0.75 0.05 26 0.74 27 Se, p11 1 0.03 rainy 0.11 0.01 0 0.14 0.01 6 0.04 0.00 50 0.10 15 dry 0.05 0.00 38 0.10 0.00 3 0.07 0.00 13 0.07 18 Fe, pl rainy 2.37 0.12 2.79 0.15 2.17 0.12 2.47 dry 2.08 0.13 2.07 0.09 2.87 0.16 2.39

Liver (D.M. basis)
Fe, PF C180 rainy 364 48.9 18 419 84.3 7 315 33.3 17 363 14 dry 519 77.6 29 482 59.6 0 291 31.9 17 409 19 Co. pp.S 75 rainy 47.8 7.78 82 35.3 6.62 93 97.7 15.7 44 62.0 72 dry 59.5 10.6 74 109.3 20.8 57 125.1 12.0 23 99, 51 Mn, pp. 6 rainy 7.69 0.48 18 12.77 1.02 0 11.45 0.56 0 10.4 6 dry 11.18 0.88 32 12.96 1.30 0 8.43 0.28 3 103 15 Zn, ppm 84 rainy 87.6 8.48 53 97.5 7.21 13 82.2 3.87 56 88.6 42 dry 83.7 8.11 74 108.2 10.4 21 93.8 3.58 27 93.3 45 Co, ppm < 0.05 rainy 0.25 0.02 0 0.27 0.04 0 0.18 0.04 0 0.23d 0 dry 0.12 0.02 35 0.12 0.02 29 0.03 0.00 87 0.08e 55
d
M. Ppm 1 4 rainy 0.63 0.13 0.56 0.09 0.50 0.04 0.56 dry 0.03 0.00 0.05 0.01 0.03 0.00 0.03' Sn, pp . 0.25 rainy -
dry 0.90 0.13 0 1.09 0.14 0 1.01 0.06 0 0.99 0 aMoDowell et al. (1984).

beans based on the following umber of samples: -rum 36, 34 (rainy, dry) for t;-1 lore farm; 31, 34 (ranv, dry) for Don Puoi- an1d 24, 46 (rainy, dry) for San Francisc. Liver 17, 23 (rainy, dry) for San Jorge farm; 15, 14 (rainy, dry) for Don Benito and 18, 30 (rainy, dry) for San Francisco.
CStandard error of means.
d""Mean. brt.-ee seasons in a o ln with different surscripts differ (P111.05).








TABLE 13. BLOOD SERUM AND LIVER MICPOMINERAL CONCENTRATIONS AS RELATED TO SEASON
AND ANIMAL CLASS.


CrltiCal Ewes Lambs Yearlings 00,ra 11 levela season MeaNb S.E.C % Def. Mean S.E. % Def. Mean S.E. % Oef. Mean % Sef.


Variable


Ser Cu, ppm Z., pp.


Se, ppm Fe, ppm Liver (D.4. basis)
Fe, ppm Cs, pp. Mn, ppm


Zn, ppm Co, ppm


Me, ppm Se, ppm


o 6 rainy 0.82 0.05 19 0.78 0.05 24
dry 1.31 0.06 0 0.99 0.05 0 0.6 rainy 0.80 0.04 11 0.78 0.03 5
dry 0.70 0.03 21 0.99 0.14 22 0.03 rainy 0.12 0.01 16 0.10 0.01 5
dry 0.07 0.01 18 0.06 0.00 37 rainy 2.47 0.13 0 2.90 0.14 0 dry : 2.61 0.18 7 2.86 0.19 0


0.85 0,03 14 1.23 0.04 0 0.69 0.02 23 0.64 0.02 33 0.09 0.01 20 0.08 0.00 7 2.21 0.12 3 2.03 0.09 2


<180 rainy 344 27.6 8 214 50.2 43 448 74.8 11 363 14
dry 301 28.5 30 913 213 0 380 32.1 16 409 19 75 rainy 56.9 11.3 80 33.2 6.63 000 80.4 12.4 50 62.0 72 dry 90.7 18.9 60 91.9 31.9 33 103.3 10] 49 99.3 51
6 rainy 10.35 0.53 4 7.96 1.31 29 11.5 0 94 0 10.4 6
dry 9.70 0.77 25 9.12 3.01 33 10.70 0.53 8 10.3 15 o 84 rainy 82.6 3.51 52 76.0 11.2 43 102.0 7 91 28 88.6 42
dry 81.5 6.79 65 116.3 26.2 33 94.5 4.03 39 93.3 45

0.05 rainy 0.26 0.03 0 0.23 0.04 0 0.18 0.02 0 0.23 0
dry 0.08 0.01 55 0.06 0.01 50 0.08 0.01 55 0.08e 55

> 4 rainy 0.51 0.06 0.65 0.32 0.59 0.04 0.56d
dry 0.04 0.00 0.01 0,00 0.03 0,00 0.03e


C 0.25 rainy
dry 0.87 0.06


0.97 0.26


1.04 0.07


aMcD-O1l1 et al. (1984).
bMeans based on the following number of samples: serr- 17, 33 (rainy, dry) for ewes, 21, 27 (rainy, dry) for lambs a-d V., 54 (arriy, dry) for yearlings; Liver 25, 20 (rainy, dry) for ewes, 7, 6 (rainy, dry) for lambs and 18, 49 (rainy, dry) for yearlings. rStandard error of means.
d'eMeans between seasons in a colon with different so rrscrijts differ (P<0.05).


0.82 18 1.20 0 0.76 14 0.74 27 0.00 15 0.07 18

2.47
2,39







selenium and zinc tended to be higher during the rainy season, and copper higher during the dry season, but none was affected (P>.05) by season.

The percentage of serum samples deficient in copper,

according to the value of .6 ppm (McDowell et al., 1984) was 18% during the rainy season and 0% during the dry season. Among animal classes, the deficiencies in copper were 19% for ewes, 24% for lambs and 14% for yearlings during the rainy season. Forage copper concentrations were shown to be highly deficient, especially during the dry season (94%). Serum copper concentrations are often directly affected by dietary copper intakes (Rowlands, 1980). However, serum copper concentrations do not always reflect dietary copper levels and copper deficiency may occur when serum copper concentrations are high. As discussed by Underwood (1981), copper deficiency is often caused by molybdenum and sulfur, which interfere with the utilization of copper by the animal, but which may induce either low or high plasma concentrations. It has been observed that low blood copper levels are associated with a reduced microbial activity of the phagocytes in the peripheral blood in sheep and cattle and this is possibly due to a reduction in the intracellular activity of the enzyme superoxide dismutase (Grace, 1988).

Based on a critical level of .03 ppm for selenium

(McDowell et al., 1984), only 15 and 18% of the samples were considered deficient for rainy and dry seasons, respectively. There was no agreement with the percentage







deficiency found in the forage (43 and 69%) for this element.

Incidence of serum samples below the critical level of .6 ppm of zinc (McDowell et al., 1984) was 14 and 27% for rainy and dry seasons. Among animal classes, deficiencies were ewes, 11 and 21; lambs, 5 and 22; yearlings 23 and 33% for rainy and dry seasons, respectively. Because signs of zinc deficiency are nonspecific, poor zinc status should be considered in cases of unexplained reproductive problems in the ewe (Apgar and Fitzgerald, 1985). Serum zinc concentrations are a reasonable criterion to determine the status of the animal, however, values are particularly susceptible to stress of the animal during sampling and can fluctuate rapidly (Underwood, 1981).

Iron deficiency anemias are of the hypochromic

microcytic type and it has been emphasized (Underwood, 1981) that an uncomplicated iron deficiency has not been observed in cattle or sheep under normal grazing conditions.

Liver. Results of liver biopsy micromineral

concentrations as related to farm and season are presented in table 12, and as related to animal class and season are in table 13. Summary of analysis of variance of liver microminerals is shown in appendix B table 32.

There were no significant farm by animal class interactions (P>.05) for any of the liver biopsy microminerals considering seasons separately. Cobalt and molybdenum were higher (P<.05) during the rainy season.







Using the upper limit of the critical value of 75 ppm suggested for copper by McDowell et al. (1984), the percentage of liver samples deficient for this microelement were 72 for the rainy season and 51% for the dry season. However, if we use 25 ppm, which is the lower limit of the range (25-75 ppm) given by McDowell et al. (1984), the incidence of deficiencies would be 30% for the rainy season and 20% for the dry season. We can take the upper limit as the marginal level for deficiency and the lower limit as deficient. Among farms, the percentage of marginal copper deficiencies were San Jorge, 82 and 74; Don Benito, 93 and 57; San Francisco, 44 and 23% for rainy and dry seasons, respectively. Among animal classes the percent marginal copper deficiencies were ewes, 80 and 60; lambs, 100 and 33; yearlings, 50 and 49%. Undoubtedly, copper is marginal at best in a high proportion (59%) under the present conditions and special attention must therefore be given to this microelement in the formulation of mineral supplements especially in San Jorge and Don Benito.

Although the best criterion of copper status is copper content of liver, blood tests are widely used in practice (CMN, 1973). In this study soil, forage, serum and liver analyses indicated 14, 75, 8 and 59% copper deficiency, respectively. Forage appeared to aid in the diagnosis of copper deficiencies in sheep. Blood did not, but as reported by CMN (1973), if values in liver of yearlings fall







below about 25 ppm, copper concentrations in blood serum start to decrease.

For zinc, the percent liver samples below the critical level of 84 ppm suggested by McDowell et al. (1984) was 42 for the rainy season and 45% for the dry season. Among farms, deficiencies were San Jorge, 53 and 74; Don Benito, 13 and 21; San Francisco, 56 and 27% for rainy and dry seasons, respectively. There is no "best" analysis to determine zinc status in the animal (McDowell, 1985). In this study overall zinc deficiency in forages was 36%; on this basis forage appeared to aid in the diagnosis of zinc status in sheep.

Liver biopsy samples were not analyzed for selenium during the rainy season. The deficiency of selenium according to the critical value of .25 ppm (McDowell et al.,1984), was 0% for the dry season. This is in disagreement with McDowell et al. (1989) that serum and liver selenium concentrations provide good indicators of dietary selenium status in cattle. The deficiencies for selenium shown in this study were 100, 56, 16 and 0% for soil, forage, serum and liver, respectively. These data are similar to the data of Valdes et al. (1988). It can be concluded that, based on results of liver analyses and of the low percentage of deficient serum samples, selenium status of the sheep appeared to be adequate under present conditions. Forage levels, although marginally deficient, appeared to underestimate the selenium status of the animal.




Full Text

PAGE 1

MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF COLOMBIA BY RODRIGO PASTRANA 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 1989

PAGE 2

ACKNOWLEDGMENTS I wish to express my sincere gratitude and appreciation to Dr. Lee R. McDowell, adviser and chairman of my supervisory committee, for his valuable guidance and assistance throughout the investigation and preparation of the present dissertation. Acknowledgments are also due to to Dr. Joseph H. Conrad, Clarence B. Ammerman, Douglas B. Bates and Lynn E. Sollenberger for their time and advice as members of the supervisory committee. Recognition and appreciation are due to Mrs. Nancy Wilkinson and Mr. Richard Fethiere for their assistance in all laboratory work and to Dr. Frank G. Martin and Dr. Steve Linda for their assistance in statistical analysis. Special appreciation is due to Dr. Oliver Ospina of Caja Agraria and to Dr. Alfonso Naranjo of Instituto Colombiano Agropecuario (ICA) , who permitted the sample collection in order to make this work possible, to Dr. Rodrigo Lora for soil analysis and to Dr. Jorge Neira for whole blood analysis at ICA laboratories. Special recognition is due to Instituto Colombiano Agropecuario for the financial support of my studies in the United States. ii

PAGE 3

Deep appreciation goes to my wife Diana and to my five children for their companionship and encouragement during this hardship. To them, this dissertation is gratefully dedicated.

PAGE 4

TABLE OF CONTENTS Page ACKNOWLEDGMENTS ii LIST OF TABLES vii LIST OF FIGURES x ABSTRACT xi CHAPTER I INTRODUCTION 1 II LITERATURE REVIEW 3 Characterization of the Sheep Industry in Colombia 3 Mineral Status of Soil 7 Soil Acidity and Organic Matter 7 Soil Macronutrients 10 Soil Micronutrients 12 Mineral Status of Plants 16 Forage Macrominerals 18 Forage Microminerals 20 Mineral Status and Requirements of Ruminants ... 23 Tissue Macrominerals 23 Tissue Microminerals 29 Hematological Measurements 33 III MATERIALS AND METHODS 3 6 Identification and Description of Research .... 36 Sample Collection 37 Soil Samples 37 Forage Samples 39 Animal Tissue Samples 4 Whole blood and blood serum 41 Bone biopsy 4 2 Liver biopsy 4 3

PAGE 5

Page Chemical Analysis 43 Soil samples 43 Forage Samples 44 Animal Tissue Samples 45 Blood serum 45 Whole blood 45 Bone sample 46 Liver samples 46 Statistical Analysis 48 IV MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF COLOMBIA: I. MACROELEMENTS 49 Introduction 49 Materials and Methods 50 Results and Discussion 53 Soil Analyses 53 Forage Analyses 55 Animal Tissue Analyses 59 Blood serum 60 Bone 64 Correlation Coefficients of Minerals 66 Hematological Measurements 6 7 Summary and Conclusions 69 V MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF COLOMBIA: II. MICROELEMENTS 71 Introduction -,, Materials and Methods 72 Results and Discussion 75 Soil Analyses 75 Forage Analyses 78 Animal Tissue Analyses 82 Blood serum 82 Liver . , 86 Correlation Coefficients of Minerals 90 Summary and Conclusions 90 VI MINERAL CONCENTRATIONS IN LEAVES AND STEMS OF VARIOUS FORAGES OF THE COLOMBIAN PARAMO 93 Introduction 93 Materials and Methods \ 94

PAGE 6

Page Results and Discussion 96 Plant Fractions-Macrominerals, Crude Protein, and IVOMD 96 Plant Fractions-Microminerals 99 Forage Species 102 Soil-Plant Relationship 108 Summary and Conclusions 110 VII SUMMARY AND CONCLUSIONS 112 APPENDIX A FIGURES 117 APPENDIX B TABLES 121 APPENDIX C RAW DATA 146 LITERATURE CITED 180 BIOGRAPHICAL SKETCH 192 vi

PAGE 7

LIST OF TABLES Table Page 1. CONCENTRATIONS (PPM) OF MINERALS OF DRIED SOIL AND THEIR INTERPRETATION 13 2. SUMMARY GUIDE TO MINERAL REQUIREMENTS FOR RUMINANTS (DRY BASIS) 17 3. DIAGNOSIS OF SPECIFIC MINERAL DEFICIENCIES OR TOXICITIES IN SHEEP 24 4. NORMAL BLOOD HEMOGLOBIN, HEMATOCRIT AND LEUCOCYTE VALUES IN SHEEP 35 5. SOIL ORGANIC MATTER, pH, AND MACROMINERAL ANALYSES AS RELATED TO SEASON AND FARM (DRY BASIS) .... 54 6. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOMD CONCENTRATIONS AS RELATED TO SEASON AND FARM (DRY BASIS) 56 7. BLOOD SERUM AND BONE MACROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND FARM 61 8. BLOOD SERUM AND BONE MACROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND ANIMAL CLASS 62 9. HEMOGLOBIN, HEMATOCRIT CONCENTRATIONS AND LEUCOCYTE COUNTS AS RELATED TO SEASON AND FARM 6 8 10. SOIL MICROMINERAL ANALYSES AS RELATED TO SEASON AND FARM DRY BASIS) . . '. 76 11. FORAGE MICROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND FARM (DRY BASIS) 79 12. BLOOD SERUM AND LIVER MICROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND FARM 83 13. BLOOD SERUM AND LIVER MICROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND ANIMAL CLASS 84 Vii

PAGE 8

Table Page 14. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOMD CONCENTRATIONS AS RELATED TO SEASON AND PLANT PART (DRY BASIS) 98 15. FORAGE MICROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND PLANT PART (DRY BASIS) 100 16. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOMD CONCENTRATIONS BY SPECIES (DRY BASIS) 103 17. FORAGE MICROMINERAL CONCENTRATIONS BY SPECIES (DRY BASIS) 104 18. CORRELATION COEFFICIENTS BETWEEN SOIL AND FORAGE MINERALS AS RELATED TO SEASON 109 19. DESCRIPTION OF FARMS 121 20. CLIMATE AND AVERAGE MONTHLY RAINFALL (MM) 123 21. DETAILED NUMBER OF SOIL AND FORAGE COMPOSITE SAMPLES 124 22. COMPOSITION OF THE MINERAL MIXTURES 125 23. DETAILED NUMBER OF TISSUE SAMPLES 126 24. SUMMARY OF ANALYSES OF VARIANCE OF SOIL ORGANIC MATTER, PH, AND MACROMINERALS-MEAN SQUARES BY SEASON 127 25. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE CRUDE PROTEIN, IVOMD, AND MICROMINERALS-MEAN SQUARES BY SEASON 128 26. SUMMARY OF ANALYSES OF VARIANCE OF BLOOD SERUM AND BONE MACROMINERALS-MEAN SQUARES BY SEASON. . . 12 9 27. SUMMARY OF ANALYSIS OF VARIANCE OF WHOLE £LOOD VARIABLES — MEAN SQUARES BY SEASON 130 28. BLOOD SERUM AND BONE MACROMINERAL CORRELATION COEFFICIENTS AS RELATED TO SEASON 131 29. CORRELATION COEFFICIENTS BETWEEN BLOOD SERUM, LIVER, AND BONE MINERALS AS RELATED TO SEASON. . . 132

PAGE 9

Table Page 30. SUMMARY OF ANALYSES OF VARIANCE OF SOIL MICROMINERALS-MEAN SQUARES BY SEASON 133 31. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE MICROMINERALS — MEAN SQUARES BY SEASON .... 134 32. SUMMARY OF ANALYSES OF VARIANCE OF BLOOD SERUM AND LIVER MICROMINERALS-MEAN SQUARES BY SEASON . . 135 33. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE (PARTS), CRUDE PROTEIN, IVOMD, AND MACROMINERALS-MEAN SQUARES BY SEASON 136 34. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE (PARTS) MICROMINERALS-MEAN SQUARES BY SEASON 137 35. FORAGE MACROMINERAL, CRUDE PROTEIN, AND IVOMD CONCENTRATIONS AS RELATED TO SEASON AND SPECIES (DRY BASIS) 138 36. FORAGE MICROMINERAL CONCENTRATIONS AS RELATED TO SEASON AND SPECIES (DRY BASIS) 140 37. OVERALL FORAGE MINERAL CONCENTRATIONS IN DIFFERENT SPECIES (DRY BASIS) 142 38. SOIL ORGANIC MATTER, PH, MACROMINERAL AND MICROMINERAL CORRELATION COEFFICIENTS AS RELATED TO SEASON 39. FORAGE MACROMINERAL, MICROMINERAL, CRUDE PROTEIN, AND IVOMD CORRELATION COEFFICIENTS AS RELATED TO SEASON 143 144 ix

PAGE 10

LIST OF FIGURES Figure Page 1. COLOMBIA, GEOGRAPHICAL LOCATION OF THE THREE SHEEP FARMS SURVEYED 117 2. THE PARAMO REGION IN COLOMBIA 118

PAGE 11

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 MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF COLOMBIA By RODRIGO PASTRANA December 1989 Chairman: Dr. Lee R. McDowell Major Department: Animal Science A study was conducted in the paramo region of Colombia to determine the mineral status of grazing sheep by evaluating mineral concentrations in soil, plant, and animal tissues. Samples were taken at the end of the rainy season (May-June, 1987) and at the end of the dry season (February, 1988) from lactating (or pregnant) ewes, lambs, and yearlings. A total of 113 composite soil, 131 composite forage, 207 blood serum, 192 whole blood, 148 bone rib biopsy, and 113 liver biopsy samples were obtained. Percentage of soil samples that were deficient was selenium 100%, boron 83%, magnesium 70%, manganese 54%, and potassium 52%. Percentage of forage samples that were deficient was sodium 93%, cobalt 87%, copper 75%, selenium 56%, magnesium 53%, and crude protein 6%. None of the values for molybdenum were above the toxic limit of 6 ppm for sheep, and the copper: molybdenum ratio was at least 4:1. xi

PAGE 12

Iron and manganese were in excess of the requirements but below the maximum tolerable levels for sheep. Leaves had higher (P<.05) concentrations of calcium, phosphorus, magnesium, potassium, iron, molybdenum and crude protein than stems. Among soil minerals and the corresponding forage minerals, only calcium and magnesium had positive correlation coefficients (P<.05, r>|.50|) for both seasons. Serum showed phosphorus deficiency in 59% of the samples during both seasons, and calcium 94% during the dry season. Both calcium and phosphorus were deficient in 98% of bone samples. Liver indicated a copper deficiency in 59% of the samples, and of zinc in 44%; during the dry season 55% were deficient in cobalt. Differences (P<.05) among animal classes were found in serum phosphorus in both seasons as was the case for serum magnesium during the dry season and bone ash during the rainy season. In conclusion, minerals most likely limiting sheep production in the paramo were calcium, phosphorus, magnesium, copper, zinc and cobalt. Supplementation programs should provide the above minerals plus common salt and selenium. xii

PAGE 13

CHAPTER I INTRODUCTION Paramo is an ecological concept that refers to the mountainous regions of the humid equatorial Andes above the upper limit of the forest (3,100 m) ., it is characterized by extreme environmental conditions, acid soil, low atmospheric pressure, low mean temperature, and high humidity. As such, it is an unique phenomenom in our planet, which only occurs in four countries: Colombia, Venezuela, Ecuador, and Costa Rica. The great importance of the paramo in Colombia is that this region is the country's source of water; most of the rivers have their origin in these very high lands. The paramo is covered with shrubs and native grasses, and is better suited for sheep grazing rather than cattle. About 47% of the sheep population (total population: 2,464,000) are wooled sheep that graze in the mountains. However, sheep production is not very efficient. Feed is in short supply during the dry season, and in general the few forage species that grow in the paramo are considered of low quality and their regrowth is very slow after grazing. Under these conditions, the animals do not consume their nutritional requirements and produce inefficiently. The poor growth rate of lambs, low fertility in particular of

PAGE 14

2 imported breeds, high mortality, and low wool production are characteristics of sheep in the paramo of Colombia. As is known, ruminants grazing forages in severely mineral-deficient areas may be more limited by mineral insufficiency than either a lack of protein or energy. Mineral deficiencies or imbalances in soils and forages are often responsible for low production and reproduction among grazing livestock. As grazing livestock usually do not receive mineral supplementation, except for common salt, they must depend almost exclusively upon forages for their requirements. Therefore, supplemental feeding of adequate minerals, which are frequently deficient in paramo forages, may improve this problem. The objectives of this study were as follows: 1. to establish if specific mineral deficiencies or toxicities exist in the paramo region of Colombia; 2. to compare the seasonal (rainy vs dry) mineral status of soil, forages, and sheep tissues; 3. to evaluate animal tissues at different physiological states of the sheep (lactating-pregnant ewes, lambs, and yearlings) ; and 4. to study mineral soil-plant-animal interrelationships.

PAGE 15

CHAPTER II LITERATURE REVIEW Characterization of the Sheep Industry in Colombia Colombia, the northernmost of all South American republics, is located at the north east corner of South America between 4° 50' to 79° 01' west of the Greenwich Meridian (Instituto Geografico Agustin Codazzi, 1986). The capital of Colombia, Bogota, lies at an altitude of 2,640 m above sea level (Appendix A, Figure 1) . The total area of the country is 1,141,748 Km 2 and, while there is a great variety of geographical areas, the country has been divided in 5 natural regions. The Andes region and its interandean valleys include the high mountain ranges of the three Cordillera chains (Cordilleras Oriental, Central and Occidental) , the Macizo de los Andes and the Sierra Nevada. The Coastal plains, both Pacific and Atlantic, and the Savanna (Llanos Orientales) are included in the Llanuras del Pacifico, Llanuras del Caribe, and the Orinoquia regions, respectively. A dry, almost desert zone in the Peninsula de la Guajira is also included in the Caribe region. A tropical jungle is found in the Amazonia region (Instituto Colombiano Agropecuario, 1980) .

PAGE 16

In general, two different types of sheep, hair and wooled, are found in Colombia, according to two climatic regions. The hair sheep are concentrated in the lowlands with high temperatures all year, which range from 20 to 33°C under both humid and very dry conditions. Hair sheep are well adapted to tropical environments. The African sheep, as hair sheep are known in Colombia, are usually brown, ranging in shade from tan to brown and cherry-red to dark red (Bradford and Fitzhugh, 1983) . The Peninsula de la Guajira holds about 3 0% of the total sheep population, most of which are hair sheep. The total sheep numbers in the country were estimated to be 2,464,000 in 1982 (Caja Agraria, 1985). The wooled sheep are found in the mountain regions where temperatures range from 9 to 15° C; in most cases they graze the higher elevations of the mountains, called paramos, where temperatures are lower than 9°C and humid conditions persist. The departamento de Boyaca leads the country in wooled sheep numbers with about 26% of the total sheep population, followed by Cundinamarca with close to 10%. Santander and Narino are also important regions and they both have 11% of the sheep in Colombia (Caja Agraria, 1985) . The predominant type of wooled sheep is the Native or Criolla breed which is well adapted to the harsh environment of the paramo. Importations of Romney, Corriedale, Hampshire, Scottish Blackface and others have occurred in

PAGE 17

the last 30 years and substantial crossing of the Criolla has resulted especially with the Romney breed. In general, the paramo region has not been exploited in a systematic way and its utilization is in its infancy. The production capability of a sheep industry in the paramo is difficult to determine as sheep have to compete not only with technological but also social restrictions, for example, lack of roads, isolation, lack of electricity, and others (Proyecto Ovino Colombo Britanico, 1979) . Among the technical restrictions to the sheep industry in the paramo we can mention the poor growth rate of the lambs until weaning; low bodyweights at 6, 12, and 18 months of age; low fertility in particular with imported breeds; high mortality rate due to several causes; and low production of wool of inferior quality. One important restriction is the very low production of forage which can be attributed to cold weather and periods of excess rainfall or drought (Proyecto Ovino Colombo Britanico, 1979) . In a description of the region, Guhl (1982) contends that the paramo is an ecological concept that refers to the mountainous regions of the humid equatorial Andes above the upper limit of the forest (above 3,100 m) . The paramo is characterized by extreme environmental conditions, acid soils, low atmospheric pressure, high humidity, low mean temperature (with daily oscilation) , and high air dryness. It is also characterized by the elevated soil and air temperatures during the direct solar radiation and by the

PAGE 18

6 sudden and dramatic changes due to the cloudiness during the night. These factors produce frosts and heavy winds during specific times of the year. The paramo is a unique phenomenon! in our planet, which only occurs in four countries, Colombia, Venezuela, and Ecuador. In Costa Rica the climatic effect of elevations at 2,500 m above sea level are comparable to those of the paramos (Robayo et al., 1988). Very little is yet known about the complex interrelationships that keep animal and plant life and that permit the existence of the paramos. Less is known of the possible utilization of the paramo lands by man. The landscape in the paramo is not totally homogeneous, and three altitudinal zones may be distinguished: the superparamo on the higher elevations (above 4,200 m) , the proper paramo (3,800-4,200 m, and the subparamo on the lower elevations (3,100-3,800 m) (Salamanca, 1986). The paramo region can be observed in appendix A figure 2. The natural vegetation in the paramo is adapted to withstand cold and dry conditions. Although the paramo is very humid, in some cases the effective rainfall is low (less than 1,000 mm in a year). A permanent fog in combination with low temperatures keeps the relative humidity at an almost 100%. The soils are generally saturated but is difficult for the plants to utilize the water and so they are in a similar situation as if they were in an arid zone (Salamanca, 1986) .

PAGE 19

7 Apart from its possible utilization with grazing animals or other forms of agriculture, the great ecological importance of the paramo lies as a source of water. Most of the rivers in Colombia have their origin in these very high lands (Cleef, 1980). Around 70% of the Colombian population live and work on the mountains. But the mountain has several aspects, and the higher elevations, as are the paramos, become a hostile region to man for its climate and topography, and are natural obstacles to communications (Guhl, 1982). Mineral Status of Soil Soil Acidity and Organic Matter McDowell (1985) indicates that soil is the source of all mineral elements found in plants. Most naturally occurring deficiencies in livestock are associated with specific regions and are directly related to both soil mineral concentration and soil characteristics. Of the total mineral concentration in soils, only a fraction is taken up by plants. Conrad (1978) indicates that grazing ruminants are in close and continuing contact with the soils on which the plants grow. Soils often contain many times the levels of minerals which are found in the plants which grow on these soils.

PAGE 20

For Reid and Horvath (1980), the availability of minerals in soils depends upon their effective concentration in soil solution. Williams (1963) indicates that the soil content of an element would seem the most important limitation. However, availability factors including soil pH, texture, moisture content, and organic matter are probably more often the limiting factors rather than soil mineral content. Sanchez (1976) says that the majority of the soils of the humid tropics is acid. Soil acidity problems are associated with pH levels lower than 5.5 and the presence of exchangeable aluminum in the soil. Percent aluminum saturation, calculated on the basis of effective cation exchange capacity, is a useful measure of soil acidity. For Sanchez (1976) , acid soil infertility is due to one or more of the following factors: aluminum toxicity, calcium or magnesium deficiency, and manganese toxicity. Aluminum toxicity is the most common cause of acid soil infertility. This toxicity can be corrected by liming to pH 5.5 to 6.0 to precipitate the exchangeable aluminum as aluminum hydroxide. Liming rates can be calculated on the basis of 1.65 ton/ha of CaCOj equivalent per milliequivalent of exchangeable aluminum. Overliming to pH values greater than 6 or 7 can seriously decrease yields, particularly in soils high in iron and aluminum oxides (Kamprath, 1972). Concentrations of soil solution aluminum above 1 ppm often causes direct reduction in yield. Aluminum tends to

PAGE 21

accumulate in the roots and impede the uptake and translocation of calcium and phosphorus. Thus aluminum toxicity may produce or accentuate calcium and phosphorus deficiencies (Foy, 1974) . Under grazing conditions, sheep and cattle can ingest large amounts of aluminum of nonplant origin when soil is involuntarily ingested with forages. Exchangeable aluminum concentrations of 6,000 and 18,000 ppm have been reported for samples of temperate and tropical soils, respectively (Velez and Blue, 1971) . Manganese is very soluble at pH values lower than 5.5. If present in sufficient amounts, manganese toxicity can occur along with aluminum toxicity at pH values of up to about 5.5 to 6.0. Manganese is a plant nutrient; consequently, the aim is not to eliminate soluble manganese but to keep it within a range between toxicity and deficiency. A solution concentration between 1 and 4 ppm represents such a range (Black, 1967) . Referring to organic matter, Sanchez (1976) indicates that in unfertilized soils the beneficial effects of organic matter consist of supplying most of the nitrogen and sulfur to plants, maintaining cation exchange capacity, blocking phosphorus fixation sites, improving structure in poorly aggregated soils, and the formation of complexes with micronutrients. For Dahnke and Vasey (1973), a test for total nitrogen and a test for organic matter or organic

PAGE 22

10 carbon are essentially the same because the concentration of nitrogen in soil organic matter is relatively constant. Soil Macromitrients Towers and Clark (198 3) indicated that mineral concentrations in both soil and plant influence the mineral status of grazing animals but many other factors such as total dry matter intake, selective grazing, interactions among minerals, variations in mineral requirements with differences in age, sex or production level also play an important part. This means that soil analysis alone cannot be used to reliably diagnose trace element deficiencies in animals. A more satisfactory soil analysis to relate mineral concentrations for livestock rather than the concentration of a mineral in a soil, is the use of soil extracts (i.e., 0.1N HC1 or 2.5% acetic acid), which contain the more available forms of soil minerals. Analyses to determine the available forms of soil minerals can sometimes provide evidence of livestock mineral deficiencies, but more often they are unreliable and difficult to interpret (McDowell et al., 1986). Calcium is essential, not only to correct, soil acidity but also as a nutrient element necessary for normal plant growth. Soil calcium plays an essential role in regulating soil pH (Sillanpaa, 1982). The same author determined that the calcium concentration of soils are poorly reflected in the calcium concentration of plants.

PAGE 23

11 Doll and Lucas (1973) stated that plants usually contain more potassium than any other nutrient element except nitrogen. Crops utilize from 50 to over 200 kg/ha, depending upon the yield and kind of crop. Supplemental potassium fertilizers are frequently needed since most soils can not meet the requirements of continued cropping. Levels of calcium in crops are lower than potassium, and the amount needed by crops usually is between 20 and 150 kg Ca/ha. However, nutrient deficiencies of calcium are not common since most soils either contain high levels of calcium or have been supplied with ample calcium because of liming. Uptake of magnesium by crops is relatively low, from less than 10 to about 25 kg/ha. Occasional magnesium deficiencies have been noted, especially on sandy soils or soils with low levels of magnesium in relation to potassium and possibly calcium. Phosphorus is classed as one of the macronutrients, but its concentration in plants is considerably less than that of nitrogen, potassium and calcium. As a limiting factor, however, phosphorus is more important than calcium and probably more important than potassium. The forms of phosphorus that occur in soil parent materials are generally of low availability to plants. Probably all forms of phosphorus in soils are of some significance in supplying phosphorus for plants on a long-term basis, but none of the forms that have been identified are known to be of significance in the short-term relationships that are of

PAGE 24

12 importance in determining the current availability of soil phosphorus (Black, 1968) . Sanchez (1976) stated that in general, sulfur deficient soils have one or more of the following properties. They are high in allophane or oxides. They are also low in organic matter and often sandy. Soils subject to repeated annual burning are often sulfur deficient since about 75% of the sulfur is volatilized by fire. In temperate regions total soil sulfur is positively correlated with organic matter and inversely correlated with degree of weathering. Sanchez (1976) also stated that sulfur deficiency in grazing sheep and cattle is not a problem, but its effect on copper absorption and metabolism is important. Sodium is of importance in plant nutrition not only because sodium is required by at least a few plants, but also because of its relation to potassium. Sodium and potassium are the two principal monovalent metallic cations in plants, and an increase in one generally brings about a decrease in the other (Black, 1968) . Table 1 provides a summary guide to the levels of soil fertility and its interpretation according to several authors. Soil Micronutrients Sulfur and the micronutrients can be differentiated from nitrogen, phosphorus, and potassium in that they are much less frequently the limiting factor in soil fertility (Foth and Ellis, 1988) . Both copper and zinc occur in the earth's crust primarily as sulfide minerals. Deficiencies

PAGE 25

13 TABLE 1. CONCENTRATIONS (PPM) OF MINERALS OF DRIED SOIL AND THEIR INTERPRETATION 3 Levels Soil type Element Low Medium High Reference Calcium 0-71 72-140 >141 b Magnesium 0-30 30-50 > 60 c Phosphorus 9-17 17-35 35-70 OrganicMineral c Potassium 31-62 62-124 124-248 OrganicMineral c Sulfur <10 >10 d Boron < .4 e Copper < .1-.3 pH 5.5-6.0 c Iron < 2.5 > 4.5 f Manganese < 3-5 Zinc < .5 pH 5.5-6.0 pH 5.5-6.0 For grazing livestock soil concentrations suggesting deficiencies are as follows: calcium (70 ppm) , potassium (58.5 ppm) , magnesium (8.4 ppm), phosphorus (10 ppm), cobalt (0.1 ppm), copper (0.6 ppm) , manganese (19 ppm), and zinc (2 ppm) (McDowell et al . , 1986). b Breland, 1976. c Rhue and Kidder, 1983. d Cooper, 1968. e Gammon, 197 6. Viets and Lindsay, 1973.

PAGE 26

14 of copper are not commonly found in mineral soils. Organic soils containing little ash are more likely to be deficient. More than 99% of the copper in the soil solution is complexed by organic matter. This complexing is of great importance in maintaining adequate copper in solution for plant use. Organic matter does complex zinc in soil solution, but the percentage of zinc that is complexed varies over a considerable range (Foth and Ellis, 1988) . Since manganese solubility is related to oxidationreduction reactions in the soil, the availability of manganese is closely related to weather. Cool temperatures may slow down the mineralization of organic manganese. On the other hand, cool temperatures associated with high levels of rainfall in early spring may keep more manganese available through reduction of manganese oxides. There is an interaction between manganese and iron. High levels of available iron in organic soils or high levels of organic matter in sands may lead to a manganese deficiency because a high ratio of iron to manganese is created within the plant (Foth and Ellis, 1988) . Foth and Ellis (1988) also state that few, if any, soils are deficient in total iron since the total soil iron concentration varies from 1,000 to 10,000 ppm. But the solubility of iron in soils may be limited by the low solubility of iron hydroxides and oxides in the pH range in which crops are grown. Soil conditions that lead to iron

PAGE 27

15 deficiency in plants include pH above 7.0, low soil moisture concentration, and low organic matter concentration. Boron is associated with soil organic matter, and soils with high levels of organic matter usually contain adequate boron for high soil fertility. Boron deficiency is often accentuated when soil contains little moisture. As with other elements, the total concentration of selenium in soils shows little relationship to the concentration of selenium in plants grown on those soils. This is because selenium in soils exists in several chemical forms which differ widely in their solubility and availability to plants. The chemical forms of selenium (selenides, selenites, selenates, organic selenium) are closely related to oxidation-reduction potential and pH of the soil (Lakin, 1972). Many studies have determined the factors affecting the availability of the microelements. The amount of most trace elements in herbage grown in freely drained soils is normally lower than on corresponding poorly drained soils (Swift, 1972) . As soil pH increases, the availability and uptake of iron, manganese, zinc, copper, and cobalt decrease, whereas molybdenum and selenium concentrations increase (Pfander, 1971; Williams, 1963; Latteur, 1962; Miller et al., 1972). Many data have accumulated indicating a decrease in the solubility of zinc with increasing pH (Foth and Ellis, 1988) .

PAGE 28

16 Low soil temperatures usually decrease micronutrient availability and may cause deficiency signs to develop during cold springs only to disappear when the soil warms up and more roots develop (Viets and Lindsay, 1973) . Mineral Status of Plants Of the thirteen essential minerals obtained from the soil by plants, five are used in relatively large quantities and are thus referred to as macronutrients. These are phosphorus, potassium, calcium, magnesium, and sulfur. The other minerals, iron, manganese, copper, zinc, boron, molybdenum, chlorine and cobalt, are used by higher plants in very small amounts, thereby giving them the designation of micronutrients or trace elements (Brady, 1974; McDowell et al. , 1983) . Mineral analysis of the forage consumed by the grazing animal is basic to mineral status diagnosis. If mineral concentrations are below minimum requirements or above the maximum tolerance level, there is an immediate suggestion of a nutritional problem. However, relying on a forage mineral analysis to establish mineral status assumes that the sample is representative of what animals consume. An additional disadvantage of forage element analyses is the difficulty of estimating forage intake and digestibility (McDowell et al., 1986) . Table 2 presents a guide to mineral element

PAGE 29

17 TABLE 2. SUMMARY GUIDE TO MINERAL REQUIREMENTS FOR RUMINANTS (DRY BASIS) Element Requirement 8 Calcium .18-. 60 % Phosphorus .18-. 43 % Magnesium .04-. 18 % Potassium .60-. 80 % Sodium . 10 % Iron 10-100 ppm Copper 410 _ , ,^ ... ppm Cobalt Zinc Manganese Molybdenum "Summarized by McDowell et al. (1978) .05-. 10 ppm 1050 ppm 2040 ppm .01 ppm or less

PAGE 30

18 requirements for grazing ruminants, dry basis, summarized by McDowell et al. (1978). Gross and Jung (1981) established that forages supply much of the minerals in diets fed to cattle, and that variation in the amount of minerals in forages are associated with season, fertilization, soil type, and soil pH. Both amount of mineral in forages and biological availability of minerals need to be considered in formulating rations. Although mineral concentration of forages can be determined chemically, biological availability is much more difficult to estimate. Biological availability of mineral elements in forages is probably a partial function of the extent to which they are solubilized in rumen fluid (Kincaid and Cronrath, 1983) . Grazing livestock usually do not receive mineral supplementation except for common salt and must depend almost exclusively upon forages to meet their requirements. Only rarely, however, can forages completely satisfy all mineral requirements (McDowell et al., 1982). Forage Macrominerals Phosphorus deficiency is found frequentlyin tropical grazing areas around the world (Cohen, 1980) , and may be the first limiting mineral deficiency under many grazing conditions. Aluminum and iron in ingested soil may interfere with dietary phosphorus utilization (Rosa et al.,

PAGE 31

19 1982) and this effect could be critical if the animals were in a borderline phosphorus deficiency. The phosphorus requirement of a ruminant is rarely met by forage diets; therefore, supplementation is then necessary (Cohen, 1980) . Calcium deficiency is a lesser problem than phosphorus deficiency in grazing animals because most herbage contains adequate calcium, and maturity has only a small effect on calcium concentration. In contrast, most of the world's rangeland soils are low in phosphorus and support herbage of low phosphorus concentration which declines markedly with maturity (Underwood, 1981) . Magnesium occurs widely in plant and animal tissues. Magnesium content of most grazed herbages usually exceed 0.1% so requirements for supplemental magnesium in grazing cattle and sheep are usually associated with induced hypomagnesemia as a result of higher concentrations of nitrogen and potassium due to fertilization rather than low magnesium levels per se (Cohen, 1987). Metson et al. (1966) suggested that magnesium levels of 0.25% were necessary to prevent grass tetany when concentrations of nitrogen and potassium are high. Potassium is often associated with high levels of nitrogen and moisture in lush, cool-season grasses (Boling et al., 1979). Ruminants grazing pasture heavily fertilized with potassium have developed hypomagnesemic tetany, but adding potassium salts directly to their diet usually failed to produce this condition (Tomas and Potter, 1976) .

PAGE 32

20 Excess sodium salts in the diet may increase the rate of passage of digesta and, thus, enable a greater quantity of dietary protein to escape ruminal degradation (Reffett and Boling, 1985) . Moseley and Jones (1974) reported that feeding high levels of NaCl resulted in increased magnesium absorption, but concomitantly resulted in increased urinary excretion of sodium and magnesium. Forage Microminerals An accurate determination of zinc requirements of ruminants is not available, although the level of 25 to 30 ppm in forage is consistent with results obtained from grazing experiments. Zinc concentrations may be 30 ppm in herbage and occasionaly higher, but this concentration declines rapidly as plants mature and values can decrease to less than 15 ppm (Mayland et al., 1987). The bioavailability of zinc may be reduced by cellulosic binding (Bremner and Knight, 1970) . Forage copper concentrations vary from 4 to 5 ppm to values of 10 to 15 ppm. Sulfur and molybdenum interfere with the absorption of copper. Variations in copper absorption may more often be associated with changes in soil pH or redox potential that have affected the solubility and uptake of molybdenum and/or sulfur (Langlands et al., 1981; Lesperance et al . , 1985). Absorption of copper varies according to age and dietary factors such as the level of molybdenum and sulfur. Zinc and calcium levels in the diet may also interact with

PAGE 33

21 copper absorption, but to a lesser extent than do molybdenum and sulfur. Herbage containing 5-6 and 7-10 ppm of copper should meet the copper requirement of sheep and cattle, respectively, unless amounts of molybdenum and sulfur intake are high. A copper: molybdenum ratio of 2.0 or greater is desirable to avoid molybdenosis (Miltmore and Mason, 1971; Ward, 1978) . A deficiency of copper in cattle occurs when the dietary level is much less than 5 ppm DM or when molybdenum or sulfur are in excess (Ward, 1978) . Ward (1978) reported that dietary copper: molybdenum ratios less than 2:1 contribute to a copper deficiency, whereas an excess of dietary sulfur potentiates the effect of molybdenum. Cobalt is the metal cofactor in vitamin B 12 which is in turn required in energy metabolism in ruminants (Mayland et al., 1987). Pasture herbage levels of at least 0.08 and 0.11 ppm will provide adequate cobalt for cattle and sheep, respectively (Grace, 1983) . Ruminants on Phalaris pastures can develop an acute form of a disease from which they quickly die or a chronic form of a nervous disorders characterized by muscle tremors, rapid breathing and pounding heart beat. Phalaris staggers is the name given to this disorder. Grazing Phalaris increases the cobalt requirement (Mayland et al., 1987). The toxicity and metabolism of molybdenum are dependent not only on the levels of dietary molybdenum but also on the levels of other dietary components. The higher the level

PAGE 34

22 of molybdenum, the greater the amount of copper required to prevent signs of molybdenosis. Prolonged high molybdenum intakes cause hypocupremia (Underwood, 1981) . Molybdenum-induced copper deficiency is an endemic problem in ruminants (Ward, 1978). Dietary levels of molybdenum are affected by parent soil material, soil pH, forage type and forage maturity (Reid and Horvath, 1980) . Molybdenum combines with hydrogen sulfide in the rumen to form thiomolybdates which render copper unavailable for absorption (Dick et al., 1975). In many areas sheep and cattle grow normally on pastures containing 0.03 ppm of selenium and show no evidence of a deficiency. In other areas, white muscle disease can occur, especially in lambs, where pastures contain as much as 0.05 ppm. These responses may be attributed to variations in dietary sulfur levels or other factors that affect the absorption of selenium or the requirement of selenium by the animal. The requirement for selenium in sheep is given by the National Research Council (NRC, 1985) as 0.1-0.2 ppm. A value of o.l is often used as a critical level, but this should be evaluated on a basis of animal performance (Mayland et al . , 1987). Selenium deficiency is associated with pastures that contain less than 0.03 ppm (Millar, 1983).

PAGE 35

23 Mineral Status and Requirements of Ruminants Table 3 presents the requirements and critical levels for the diagnosis of specific mineral deficiencies or toxicities in sheep. Without question, forage analysis is a much better indicator of mineral status for ruminants than is soil analysis. Likewise, animal tissue-mineral concentrations are better indicators of the availability of minerals than are forage mineral analyses. Grazing livestock obtain part of their mineral supply from the consumption of water, soil, leaves, tree bark, etc, rather than entirely from forages (McDowell et al., 1986). Evaluating the mineral status of domestic animals can be a complex and costly procedure. Often little is known about the nutritional background of animals in question. Collecting blood or tissue samples for mineral analysis is often impractical because of limited facilities and personnel trained to collect samples (Combs, 1987) . Tissue Macrominerals Calcium is the most abundant of the minerals in the animal body. About 2 6-3 0% of total ash content of most animals is calcium. Although about 98-99% of the total body calcium is in the skeleton, it has numerous crucial functions in soft tissues. The active form of calcium in soft tissues is the ionized form. Ionized calcium content of blood plasma is homeostatically regulated within

PAGE 36

24 TABLE 3. DIAGNOSIS OF SPECIFIC MINERAL DEFICIENCIES OR TOXICITIES IN SHEEP Element Dietary requirement (dry basis) 3 Tissue Critical level b,c Deficiency Calcium .20-. 82 % Bone (fat free) Bone ash Plasma 24.5 % 37.6 % 8 mg/dl Phosphorus .16-. 38 % Bone (fat free) Bone ash Plasma 11.5 % 17.6 % 4 . 5 mg/dl Magnesium .12-. 18 % Serum Urine 1-2 mg/dl 2-10 mg/dl Potassium .50-. 80 % Sodium .09-. 18 % Saliva 100-200 mg/dl Sulfur .14-. 26 % Iodine .10-. 80 % Milk 300 ug/day Iron 30-50 ppm Hemoglobin Transferrin 10 g/dl 13-15 % saturation Copper 7-11 ppm Liver Serum 25-7 5 ppm . 65 ug/ml Molybdenum .5 ppm Cobalt .1-.2 ppm Liver .05-. 07 ppm Manganese 2 0-4 ppm Liver 6 ppm Selenium .1-.2 ppm Liver Serum Hair or wool . 2 5 ppm . 03 ug/ml .25 ppm

PAGE 37

25 TABLE 3. -CONTINUED Element Dietary requirement (dry basis) 3 Tissue Critical level Toxicity Copper Fluorine Manganese Iron Molybdenum Selenium Zinc 25 ppm 60-150 ppm 1000 ppm 500 ppm 10 ppm >2 ppm 750 ppm Liver Bone Hair Liver Liver Hair Hair 700 ppm 4500-5500 ppm 70 ppm 4 ppm 5-15 ppm 10 ppm 10 ppm "NRC (1985) . Requirements below which a deficiency occurs. References for critical levels are found in the following reviews: Mtimuni (1982); McDowell et al. (1984). c Non-mineral assays for the following elements are sensitive diagnostic techniques: cobalt (Vitamin B, 2 ) , iodine (free thyroxine) , copper (ceruloplasmin) and selenium (glutathione peroxidase) .

PAGE 38

26 relatively narrow limits. The skeletal system serves as a very effective reservoir of calcium, which maintains ionized plasma calcium levels within narrow limits under a wide range of dietary calcium intakes (Combs, 1987) . Calcium, phosphorus and magnesium are important components of bone, and also intracellular and extracellular fluids of the body. Extracellular calcium is essential for maintenance of nerve tissue, resting membrane potential, blood clotting mechanisms, myocardial contraction and myoneural junctional transmission. Intracellular calcium, directly or indirectly, regulates activity of many enzymes, microtubule assembly, generation of ATP, release of hormones and neurotransmitters and muscle cell contraction (Littledike and Goff , 1987) . Excess calcium may impair reproductive function by causing a secondary deficiency of phosphorus, magnesium, zinc, copper and other microelements by inhibiting their absorption in the intestine (King, 1971) . Phosphorus has long been recognized as a major essential nutrient for ruminants. Approximately 80% of body phosphorus occurs in bone and skeletal development depends upon an adequate supply of phosphorus. Phosphorus is required for phosphorylation in sugar metabolism, intracellular energy transfer, formation of phospholipids, as a buffer in blood and other fluids and is required for proper functioning of rumen microorganisms (Cohen, 1987) .

PAGE 39

27 Phosphorus is also abundant in animal tissues, accounting for 16-17% of total body ash. Severe deficiencies of phosphorus in cattle reduce feed intakes, feed efficiencies, and retard growth (Kincaid et al., 1981). The requirement for phosphorus as a percent of dry matter in rations for sheep is given by the National Research Council (NRC, 1985) as 0.16-0.38%. Dietary calcium: phosphorus ratios may affect reproductive performance. Phosphorus deficiency induces lowered conception rate, irregular estrus, decreased ovarian activity, increased incidence of cystic follicles, and generally depressed fertility. When phosphorus levels are low, phosphorus supplementation, while expensive, is critical to animal performance (Hurley and Doane, 1989) . There has been increasing acceptance of the rib bone biopsy technique (Little, 1972) as a more reliable method for the estimation of phosphorus status of grazing cattle and sheep than blood. McDowell (1985) has indicated that this technique is being used widely as a survey technique to locate mineral deficiencies in tropical regions. About 70% of total body magnesium in livestock occurs in the bones. It also occurs in high concentrations in intracellular fluid where it is associated with the mitochondria and in lesser concentrations in the extracellular fluid. It is involved in oxidative phosphorylation, phosphate transfer, metabolism of carbohydrates and lipids and in neuromuscular activity

PAGE 40

28 (Cohen, 1987). Magnesium accounts for 1-1.1% of total ash of most animal species and is deposited primarily in skeletal and muscle tissue. Seven to eight % of the total body stores of magnesium is in other tissues and body fluids (Combs, 1987) . Magnesium is an essential cof actor of many enzymes, especially phosphate-transferring enzymes involved in ATP generation and the adenylate and guanate cyclases, and is essential for normal function of nerve tissues (Littledike and Goff , 1987) . The magnesium status of livestock can be assesed from plasma, whole blood or urine (Egan, 1980) . Normal magnesium levels in plasma of sheep are listed as 1.8-2.0 mg/dl with values below 1.0 ig being severely hypomagnesemic (NRC, 1985). The National Research Council (1985) suggested minimum magnesium requirements of 0.12, 0.15 and 0.18% of dry matter for growing lambs, for ewes in late pregnancy, and for ewes in early lactation, respectively. Studies have shown that excess magnesium intake by ruminants caused loss in weight, drowsiness and changes in blood mineral levels (Gentry et al., 1978). Hypomagnesemic tetany is a problem in ruminants managed under a variety of regimens. The disorder has been reported in milk-fed calves, and in adult sheep and cattle fed highroughage diets or maintained on sparse pasture (Giduck and Fontenot, 1987). A deficiency of magnesium in ruminants may result from low magnesium concentrations in feeds or a

PAGE 41

29 reduction in biological availability of dietary magnesium (Fontenot, 1982) . Sodium, potassium and chlorine function in maintaining acid-base balance, osmotic pressure and body fluid balance. Sodium is involved in transmission of nervous impulses and occurs largely in the body fluids and bones while potassium occurs mainly in the muscle, nervous tissue and erythrocytes (Cohen, 1987) . Sodium deficiency leads to a pica or craving for salt, unthrifty appearance, loss of appetite, weight loss and reduced milk yield. These signs usually occur without a decline in plasma sodium, although urinary and fecal sodium may decline (Underwood, 1981) . The National Academy of Sciences (NRC, 1985) listed sodium requirements at 0.090.18% dry matter of the diet and potassium at 0.50-0.80% dry matter. Tissue Microminerals Zinc occurs widely and in relatively high concentrations throughout the body. Zinc concentrations in plasma of sheep and cattle range from 0.6 to 1.2 ppm, respectively. Zinc is a constituent of a large number of metallo-enzymes involved in biochemical processes essential to nucleic acid and carbohydrate metabolism, as well as protein synthesis. It is associated with appetite, growth, male sexual development and wound healing. There are no significant stores of body zinc, and the animal must rely on a daily supply to meet requirements (Mayland et al . , 1987).

PAGE 42

30 Largely on the basis of Australian studies, Underwood (1981) concluded that zinc requirements for optimum growth and fertility of sheep must lie close to 30 ppm of the dry diet. The NRC (1985) requirements of sheep are given as 29 to 33 ppm. Clinical signs of severe zinc deficiency have been reported in ruminants under practical conditions. However, a marginal zinc deficiency appears to be a more widespread occurrence (Spears, 1989). Low zinc intake throughout pregnancy has severe effects on reproduction in the ewe. Because signs of zinc deficiency are nonspecific, poor zinc status should be considered in cases of unexplained reproductive problems (Apgar and Fitzgerald, 1985) . Excess dietary iron can affect performance adversely in ruminants. High dietary iron can affect utilization of other minerals such as copper, phosphorus, zinc and manganese (Humphries et al., 1983). The liver is the primary storage organ of body copper stores, having about 40-70% of the total copper. Copper is active in the conversion of tyrosine to melanin, which provides the color pigment in hair and wool. About 2 0% of the plasma copper is in a loosely bound form, while the other 80% is associated with a protein called ferroxidase I (Ceruloplasmin) . Ferroxidase I oxidizes ferrous iron (Fe 2 *) to ferric (Fe 3 *) allowing the mobilization of iron stores (Mayland et al., 1987).

PAGE 43

31 Adequate maternal intake of copper is essential for development of the central nervous system of the embryonic lamb. Enzootic ataxia of the unborn or the unweaned lamb is primarily from copper deficiency (Hidiroglou and Knipfel, 1981) . Visible signs of copper deficiency are not usually seen in the adult sheep. Copper deficient lambs may have a degenerative disorder known as swayback (Poole, 1982) . Physiological copper deficiencies are produced by four classes of feeds: (l) high molybdenum, generally above 100 ppm, (2) low copper: molybdenum ratio, 2 : 1 or less, (3) copper deficiency, below 5 ppm, and (4) high protein, 2930% protein in fresh forage (Ward, 1978) . Copper toxicity is essentially a problem of the housed ruminant because housed ruminants are given foodstuffs of high copper availability. Conversely, copper deficiency is a problem of the grazing animal because of the poor availability of copper in grass, so low as to be less than 10% of that found in some foodstuffs (Suttle, 1986) . Sulfur in the inorganic form, which is converted within the gastrointestinal tract to certain amino acids and Bvitamins by rumen microorganisms, can meet the needs of ruminants for sulfur. This process provides methionine, thiamin and biotin, which otherwise would need to be provided by the diet to meet the needs of sulfur-containing compounds for the animal's metabolic, regulatory and structural functions. All other sulfur-containing compounds

PAGE 44

32 required by mammalian tissue can be synthesized from methionine (Goodrich and Thompson, 1981) . Elevated molybdenum intakes depress copper availability and may produce a physiological copper deficiency in ruminants. Total sulfur or sulfate in the ration generally potentiates the effect of molybdenum. The ratio of copper to molybdenum in feed is important regardless of the absolute amount of each. For this reason, and because of the importance of the sulfur content of the diet, it is impossible to define safe dietary limits of copper and molybdenum (Ward, 1978) . Thiomolybdates reduce the absorption of dietary copper in sheep. They also affect systemic copper metabolism by changing markedly the distribution of copper in plasma and by reducing the availability of copper to metabolic sites within the body (Gooneratne et al., 1989). According to Mayland et al. (1987) loss of appetite, high aspartic aminotransferase, and low blood plasma glucose levels are the best indicators of cobalt deficiency. Serum vitamin B 12 less than 0.20 ng/ml will also indicate a possible cobalt deficiency, but liver cobalt levels are not always reliable estimates of cobalt status. Selenium is widely distributed in the body. The kidney and liver normally have the highest concentrations. Selenium is an integral part of the enzyme glutathione peroxidase which catalyzes the reduction of peroxides, thereby protecting tissues against oxidative damage.

PAGE 45

33 A dietary concentration of 0.1-0.2 ppm of selenium has been accepted by the NRC (1985) as a safe and adequate level for the prevention of white muscle disease in sheep. Workers in New Zealand found that lambs grew normally with no signs of white muscle disease when pastures contained 0.03 to 0.04 ppm of selenium (Hartley and Grant, 1961). However, Whanger et al. (1978) reported white muscle disease in lambs fed diets containing 0.1 ppm of selenium. High levels of oral copper have been reported to reduce selenium availability in nonruminants . Copper often is added to mineral mixes for ruminants, and copper toxicosis is relatively common in sheep (Underwood, 1981) . White et al. (1989) showed that copper or molybdenum supplements at 10 mg/kg to practical-type diets of ewes and lambs had no effect on selenium status. Hematological Measurements Blood is the principal fluid transport system in the body and provides an expedient source of metabolic products. Chemical analysis of serum and plasma provide quantitative estimates of physiological parameters for diagnosis of disease. Quantitative differences among normal cows have been found for many of these variables by Peterson et al. (1982) . Hemoglobin is the oxygen-carrying compound contained in red blood cells (RBC) . The amount of hemoglobin per 100 ml

PAGE 46

34 of blood can be used as an index of the oxygen-carrying capacity of the blood. Total blood depends primarily on the number of RBCs (the hemoglobin carriers) but also, to a much lesser extent, on the amount of hemoglobin in each RBC (Ravel, 1989). Reference values are most frequently quoted in sheep as 9-15 g/dl (Table 4). Since whole blood is made up essentially of RBC and plasma, the percentage of packed RBCs after centrifugation gives an indirect estimate of the number of RBCs/100 ml of whole blood. Hematocrit thus depends mostly on the number of RBCs, but there is some effect (to a much lesser extent) from the average size of the RBC (Ravel, 1989). White blood cells (WBC or leucocytes) form the first line of defense of the body against invading microorganisms. Neutrophils and monocytes respond to phagocitosis; lymphocites and plasma cells apparently produce antibodies (Ravel, 1989).

PAGE 47

35 TABLE 4. NORMAL BLOOD HEMOGLOBIN, HEMATOCRIT AND LEUCOCYTE VALUES FOR THE SHEEP Range 8 Average Hemoglobin, g/dl 9-15 11.5 Hematocrit, % 27-45 35 Leucocytes, /ul 4000-12,000 8000 Neutrophil (mature) , % 10-50 30 Lymphocyte , % Monocyte, % 40-75 06 62 2.5 Eosinophil, % 0-10 5 Basophil, % 03 0.5 "Schalm and Nemi (1986). /•

PAGE 48

CHAPTER III MATERIALS AND METHODS Identifica tion and Description of Research The research for the present study was conducted at three sheep farms located in the paramo region of the Cordillera Oriental of Colombia during both the rainy and dry seasons . San Jorge farm belongs to Instituto Colombiano Agropecuario (ICA) and is located in Soacha (Cundinamarca) , on one dry area of the paramo. Don Benito and San Francisco farms belong to Caja de Credito Agrario Industrial y Minero (Caja Agraria) and are located in Zipaquira (Cundinamarca) and Ventaquemada (Boyaca) , respectively, on wet areas of the paramo (Appendix B, table 20) . About 36% of the total sheep population, or 85% of the wooled sheep of the Country are located in these two departments (Cundinamarca and Boyaca) . Location of the farms is shown in Appendix A, figure 2. Soil, forage and animal tissue samples were collected from the three farms during the rainy and dry seasons. The two seasons were selected based on the pattern of rainfall. There are two relatively short but marked rainy seasons during the year: from mid-April to mid-June and then from 36

PAGE 49

37 mid-September to mid-November. Appendix B table 20 shows the climate and average monthly rainfall of two of the farms. The first sampling period corresponded to the end of the rainy season (May, June 1987) and the second sampling corresponded to the middle-end of the dry season (February 1988) . Identification and a general description of parameters in evaluating animal production for each farm appears in appendix B table 19. This information was provided by farm managers during the sample collection period in 1988. Some data were derived from an approximation by the manager in cases where exact information was not available. Sample Collection Soil Samples A total of 113 composite samples were collected from the three farms during both 1987 and 1988 sampling periods. Each composite sample was made up of 8-12 samples taken from predetermined areas of a paddock, totalling 4-5 composite samples from each paddock. Twenty, nineteen and twenty composite samples from different paddocks were collected from San Jorge, Don Benito and San Francisco, respectively, during 1987 (rainy season) sampling period. Twenty, fourteen and twenty composite

PAGE 50

38 samples from different paddocks were collected from San Jorge, Don Benito and San Francisco, respectively, during 1988 (dry season) sampling period. Samples of soil were taken from the top layer (20 cm) using a stainless steel bore. A soil sampling technique described by Bahia (1978) was used. Although soil samples collected during the two seasons did not come from the exact same spot, they came from the same grazing area of the farm. Based on texture, the soil of the upper part (above 3,000 m) of San Jorge is classified as loamy (about 50%) or silt loam (about 50%); in the lower part (below 3,000 m) , about 40% of the soil is clay, 40% is clay loam and sandy clay loam, and the rest is sandy clay, loam, and sandy loam. In Don Benito the predominant soil is loamy, and in San Francisco it is either loamy or silt loam. Proper identification of each paddock is in appendix C, and the detailed number of composite soil samples is in appendix B table 21. Soil and forage samples were collected one or two days before animal tissue sampjes were taken. Approximately 500 g composite soil were transferred to plastic bags and were properly identified for further analysis at the Soils Laboratory at Tibaitata, ICA, Colombia. Twenty gram subsamples were taken, transferred to small plastic bags and were properly identified for selenium analysis in the United States.

PAGE 51

39 Forage Samp ler A total of 131 composite forage samples were taken from the three farms during both the rainy season (1987) and the dry season (1988) . Twenty-eight, twenty and nineteen composite samples containing the major species of forage from San Jorge, Don Benito and San Francisco, respectively, were taken during the 1987 (rainy) collection period. Twenty-nine, fifteen and twenty composites from San Jorge, Don Benito and San Francisco were taken during the 1988 (dry) collection period. The forage species collected were: vernalgrass f Anthoxanthum odoratum) , a native cultivar of velvetgrass (Holcus lanatus L) , an imported cultivar of velvetgrass (H. lanatus basyn) , kikuyugrass ( Pennisetum clandestinum) , white clover (Trifolium repens ) , tall fescue (Festuca arundinacea ) , and orchardgrass ( Dactvlis Slomerata) . In both collections forage samples were taken from the same areas where soil samples were taken. Appendix B table 21 shows the detailed number of forage samples; proper identification of paddocks and of forage species is in appendix C. Each of the farms maintains about 3 sheep/ha/year under a rotational grazing system (appendix B table 19) . There is not a fertilization program in San Jorge; however, some paddocks in the lower part of the farm receive nitrogen (in the form of Urea) which is applied at 50 kg ha" 1 . In Don Benito and San Francisco pastures are not fertilized;

PAGE 52

40 however, some paddocks take advantage of residual fertilization as a consequence of potato cropping. Each of the composite forage samples from each paddock came from 2 0-25 individual samples of the same forage species predominating and most frequently grazed by sheep on the different areas of the farm. To avoid contamination, plastic gloves were utilized for forage collection. Only the aerial part of the forage (about 5 cm from the ground) was taken. Samples were collected in plastic bags and kept refrigerated at 5° C until further processing. Samples were then hand separated by species and within each species samples were further separated into two parts, stems and leaves. A third component of the sample was left totally unseparated and analyzed separately as "whole plant", but no comparisons were made with the stem and leaf components of the same plant. Plant parts were transferred to paper bags and oven dried at 60° C for 48 hours. After this process, the samples were ground in a hammer mill with stainless steel knives and 1 mm screen at ICA facilities in Bogota. After mixing, a 60 g sample was transferred into a plastic bag and properly identified for further chemical analysis in the Nutrition Laboratory of the University of Florida. Animal Tissue Samples Blood serum, whole blood, bone and liver samples were collected from sheep on each farm. The animals were divided

PAGE 53

41 into three classes: lactating (or pregnant) ewes, lambs (14 months of age) and yearlings (10-14 months of age). Because of timing differences in the breeding seasons, it was not possible to collect samples from lambs in San Francisco at the 1987 collection period because they were very young or not yet born; in this case their mothers were in their last trimester of pregnancy. Appendix B table 23 details the number of animal tissue (serum, whole blood, liver, and bone) samples taken. The animals in San Jorge were Criollo x Romney or Criollo x Corriedale crosses and in Don Benito and San Francisco the animals were Criollo x Blackface crosses (appendix B table 19) . During each sampling period, animals from the desired classes were selected at random from each farm. Because it was not possible to follow the animals sampled during the rainy season of 1987, other animals were randomly selected in the dry season of 1988. Animal stress and excitement were minimized during and prior to tissue sample collection. The composition of the mineral mixture given on the three farms is presented in appendix B table 22. A majority of the animals received mineral supplementation although this was not done for the entire year especially in San Jorge farm. Whole blood and blood serum. Duplicate blood samples were obtained by jugular puncture and were collected in monovette tubes (Sarstedt, West Germany) . One of them

PAGE 54

42 contained the anticoagulant NH 4 -heparin. This tube was inverted 5 times so the blood made contact with the anticoagulant but avoiding hemolysis of the red cells. The blood samples were left standing in a cool environment and sent to Laboratorio de Investigaciones Medico Veterinarias (LIMV) in Bogota (ICA, Colombia) for the determination of hematocrit, hemoglobin and total and differential leucocyte counts . The second tube was used for serum separation. The blood samples were centrifuged at 2500 rpm for 30 minutes at the respective farm. The serum samples were identified and kept frozen after centrifugation until the precipitation of serum proteins was completed at the University of Florida. Procedures and techniques for blood processing have been described by Fick et al. (1979). In total, 207 serum and 192 whole blood samples were analyzed. Bone b iopsy. A total of 14 8 bone biopsy samples were taken from the sheep. Bone biopsy samples were taken as described by Little (1972), with some modifications of the technique. A single sample was removed from the 12 th rib on the right side of the animal. The procedure performed using the same vertical incision, approximately 4 cm in length made for the liver biopsy, was used for the bone biopsy. The width of the rib in sheep is so narrow that the biopsy was taken without using a trephine; instead, a piece of bone approximately 2 cm in length was removed by cutting a section of rib using a small stainless steel bone shear.

PAGE 55

43 Bone samples were kept in individual plastic bottles containing 10% formalin for analysis at the University of Florida. Liver biopsy. A total of 113 liver biopsy samples were taken from the sheep. Samples were taken in vivo using the technique described by Fick et al. (1979) and McDowell et al. (1983) . The same animals on each farm selected for blood sampling were used for liver biopsy sampling. Liver samples were kept frozen in 10% formalin for further analysis at the University of Florida. Chemical Analysis Soil Samples Soil samples were analyzed by standard methods in the Laboratorio Nacional de Suelos in Tibaitata, ICA, Bogota for organic matter (OM) , pH, aluminum, boron, calcium, copper, iron, magnesium, manganese, phosphorus, potassium, sodium, and zinc. Aluminum (exchangeable acidity) was extracted from the soil samples with the solvent KC1, IN; exchangeable basis were extracted with ammonium acetate and organic matter was determined by the Walkley and Black colorimetric method (Black, 1967). Phosphorus was measured using the method of Bray II (Jackson, 1958).

PAGE 56

44 Soil mineral concentrations were determined by atomic absorption spectrophotometry (Perkin-Elmer, 1980) . Selenium was analyzed at the University of Florida, by a fluorometric method (Whetter and Ullrey, 1978) . Forage Sainp lps Forage samples were processed and analyzed for mineral concentration according to methods described by Fick et al. (1979) at the Nutrition Laboratory, University of Florida. Calcium, copper, iron, magnesium, manganese, potassium, sodium and zinc were analyzed by atomic absorption spectrophotometry in a Perkin-Elmer AAS 5000 (Perkin-Elmer, 1980) . Cobalt and molybdenum were analyzed by f lameless atomic absorption spectrophotometry using a Perkin-Elmer AAS Zeeman/3030 (Perkin-Elmer, 1984). Phosphorus was determined by the colorimetric method described by Harris and Popat (1954) and included by Fick et al. (1979) as a method for phosphorus determination for plant and animal tissues. Selenium in forage samples was analyzed by a fluorometric method (Whetter and Ullrey, 1978) . For nitrogen analysis, samples were digested using a modification of the aluminum block digestion procedure of Gallaher et al. (1975). Sample weight was 0.3 g, catalyst used was 3.2 g of 9:1 K.SO, : CuS0 4 , and digestion was conducted for 4 h at 400° C using 10 ml H 2 S0 4 and 2 ml H 2 2 . Ammonia in the digestate was determined by semiautomated colorimetry (Hambleton, 1977). Multiplication of nitrogen by 6.25 was the procedure for calculating crude protein. In

PAGE 57

45 vitro organic matter digestion (IVOMD) was performed by a modification of the two-stage technique (Moore and Mott, 1974). Dry matter was determined by drying for 15 h at 105° C and organic matter by ashing for 15 h at 550° C. Animal Tissue Samp lps Blood serum. Serum brought to the University of Florida was deproteinized with 10% trichloroacetic acid (TCA) and then analyzed for mineral content according to the method described by Fick et al. (1979). Calcium, copper, magnesium, and zinc were analyzed by atomic absorption spectrophotometry using a Perkin-Elmer AAS 5000 (PerkinElmer, 1980) . Phosphorus was determined by a colorimetric technique (Harris and Popat, 1954). Selenium was analyzed using the method of whetter and Ullrey (1978) . Whole blood. Blood samples which contained anticoagulant were used to determine hematocrit, hemoglobin, and total and differential leucocyte counts in Laboratorio de Investigaciones Medico Veterinarias (LIMV) , ICA, Colombia. The microhematocrit method was utilized using capillary tubes of 1.1-1.2 mm diameter in an International centrifuge with standardized reading scales. Centrifugation was for 5 minutes at 2500 rpm. After centrifugation, the height of the RBC column was measured and compared with the height of the column of original whole blood. The percentage of RBC mass to original blood volume is the hematocrit (Ravel, 1989).

PAGE 58

46 The hemoglobin concentration was obtained utilizing the oxyhemoglobin method. A .025 ml aliquot of blood is added to a test tube containing 5 ml sodium bicarbonate (.1% solution) and mixed (Schalm and Nemi, 1986). A Leitz spectrophotometer was utilized. To determine total leucocyte counts, a Coulter Counter model FN (Coulter Electronics, Inc, Hialeah, Florida) was used. The Coulter Counter is based on the principle that cells are poor electrical conductors. A measured volume of diluted suspension of cells (in an electrically conductive medium) is drawn through a minute aperture between two electrodes. Each cell passing through the aperture displaces an equal volume of the electrolyte solution and is counted electronically and displayed on the digital readout (Schalm and Nemi, 1986). Differential leucocyte counts were appraised visually from prepared slides of blood smears using the methods described by Schalm and Nemi (1986). Bone samples. The samples were dried and extracted in ether following procedures outlined by Fick et al . (1979) and subsequently analyzed for calcium, magnesium, and phosphorus . Liver samples . Livbiopsy sample preparation was carried out as described by Fick et al. (1979). Dry tissue samples (approximately .3 g) were pre-ashed on a hot plate with 50% (v/v) nitric acid and then ashed overnight in a muffle furnace at increments of 100° C every hour until reaching 550° c. Ash was solubilized first with 50% nitric

PAGE 59

47 acid, then with 10% nitric acid and finally, with distilled water. Solutions were filtered, diluted to appropiate range and analyzed with atomic absorption spectrophotometry for copper, iron, manganese, and zinc using a PerkinElmer AAS 5000 (Perkin-Elmer, 1980) . Liver cobalt and molybdenum were determined by flameless atomic absorption spectrophotometry using a Perkin-Elmer AAS Zeeman/3030 (Perkin-Elmer, 1984) . Selenium analysis was carried out by the technique of Whetter and Ullrey (1978) . Statistical Analysis Data were analyzed by use of the Statistical Analysis System (SAS Institute., 1985). Probability level for significance was .05 in all statistical analyses. Blood Serum. Wh ole Blond. Bone and Liver Serum, whole blood, bone and liver were analyzed as a split-plot design with animal class as the main plot and season as the subplot. The model was as follows: V ijk = u + A, + Bj . + C,j + D k + F kj + G kj + E jjk , where: u = overall mean. A, = random effect of the i th farm. Bj = effect of the j th animal class. Cij = effect of farm*animal class (experimental error for animal class) . D k = effect of the k th season. F ki = effect of season*farm.

PAGE 60

48 G kj effect of season*animal class. E (jk = residual error (experimental error associated with the subplot) . Since the data were unbalanced, hypothesis testing was based on the Type IV Sum of Squares. Soil and Forag p Soil and forage were analyzed as a split-plot design, with farm as the main plot and season as the subplot. For forage, plant part was considered as the subplot. The model was as follows: Y ,jk = u + A f + B j(i) + C k + D ik + E ijk , where: u = overall mean. A s = fixed effect of i th farm. B j(i) " effect of j th paddock within i th farm (experimental error for farm) . C k = effect of k th season. (k th plant part, in forages). D ik " effect of farm*season. (farm*plant part, in forages) . E ijk = experimental error associated with subplots. Correlation coefficients between soil and forage, forage and animal tissues, and serum, liver and bone responses were estimated. These estimates were obtained for each farm and class separately for the rainy and dry seasons. Only those correlation coefficients of biological importance are discussed in the corresponding chapters.

PAGE 61

CHAPTER IV MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF COLOMBIA. I. MACROELEMENTS Introduction Sheep are adapted to the cold and harsh environment of the Colombian paramo; but it is commonly accepted that the few forage species that are grown are of low quality and their regrowth is very slow after grazing. Under these conditions animals do not get their nutritional requirements and, therefore, the sheep enterprise is not very efficient. Poor growth rate of lambs, low fertility in particular of imported breeds, high mortality and low wool production of inferior quality are characteristics of sheep production in the Colombian paramo (Proyecto Ovino Colombo Britanico, 1979) . Mineral deficiencies, imbalances, and toxicities have been reported to severely inhibit tropical cattle production systems (McDowell, 1985). It has been demonstrated that, with the exception of calcium and sulfur, none of the macroelements have adequate concentrations in some forage species grazed by sheep on the paramo (Laredo et al., 1989). The objectives of this study were to evaluate the macromineral status of grazing sheep and to determine macroelement status and other soil, forage, and blood 49

PAGE 62

50 parameters in three farms as related to the wet and dry seasons of the Colombian paramo. Materials and Methods Soil, forage, and animal tissue samples were collected from three sheep farms in the paramo of the Cordillera Oriental of Colombia. Sampling periods corresponded to the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988). A total of 113 composite soil, 131 composite forage, 207 serum, 192 whole blood, and 148 rib bone biopsy samples were obtained for each of the sampling periods. Each composite soil sample came from 8-12 samples. A soil sampling technique described by Bahia (1978) was used. Soil samples were analyzed by standard methods in the Laboratorio Nacional de Suelos of ICA in Bogota for organic matter, pH, aluminum, calcium, magnesium, phosphorus, potassium and sodium. Minerals were extracted from the soil samples with an acid extracting solution (.025N H 2 S0 4 and .05N HC1) . Based on texture, the soil of the upper part (above 3,000 m) of San Jorge is classified as loamy (about 50%) or silt loam (about 50%) ; about 40% of the soil of the lower part (below 3,000 m) is clay, 40% between clay loam and sandy clay loam; the rest is sandy clay, loam, and sandy

PAGE 63

51 loam. In Don Benito the predominant soil is loamy, and in San Francisco it is either loamy or silt loam. Each composite forage sample came from 20-25 samples of the same forage species predominating and most frequently grazed by sheep in the different areas of the farms. Forage species collected were: vernalgrass (A. odoratum) , native velvetgrass (H. lanatus L) , imported velvetgrass (H. lanatus basyn) , kikuyugrass (P. clandestinunO , white clover (T. repens) , tall fescue (F. arundinacea l , and orchardgrass (D. glpmerata) . Each of the farms maintains about 3 sheep/ha/year under a rotational grazing system (appendix B table 19) . There is not a fertilization program in San Jorge; however, some paddocks of the lower part receive nitrogen (in the form of Urea) at 50 kg ha" 1 year. In Don Benito and San Francisco pastures are not fertilized; however, some paddocks take advantage of residual fertilization as a consequence of potato cropping. Forage samples were processed and analyzed for mineral content according to methods described by Fick et al. (1979). Samples were collected from animals that were divided into three classes: lactating (or pregnant) ewes, lambs (14 months of age) and yearlings (10-14 months). The sheep were Criollo x Romney or Criollo x Corriedale in San Jorge and Criollo x Blackface in Don Benito and San Francisco. Duplicate blood samples were obtained by jugular puncture and were collected in monovette tubes (Sarstedt,

PAGE 64

52 West Germany) . One tube contained the anticoagulant NH 4 heparin. The second tube was used for serum separation. Bone biopsy samples were taken as described by Little (1972), with some modifications of the technique. The biopsy was taken without using a trephine; instead, a piece of bone approximately 2 cm in length was removed by cutting a section of rib using a small stainless steel bone shear. Forage calcium, magnesium, potassium and sodium, serum calcium, magnesium, copper and zinc and bone calcium and magnesium were determined by atomic absorption spectrophotometry using a Perkin-Elmer AAS 5000 (PerkinElmer, 1980). Forage, serum and bone phosphorus were determined by the colorimetric method of Harris and Popat (1954). For nitrogen analysis, forge samples were digested using a modification of the aluminum block digestion procedure of Gallaher et al. (1975). In vitro organic digestibility (IVOMD) was performed by a modification of the two-stage technique (Moore and Mott, 1974). Whole blood samples were processed at the Laboratorio de Investigaciones Medico Veterinarias (LIMV) of ICA in Bogota for hematocrit, hemoglobin and total and differential leucocyte counts. Hemoglobin concentration was obtained utilizing the oxyhemoglobin method, and to determine total leucocyte counts, a Coulter Counter model FN was utilized (Schalm and Nemi, 1986). Data were analyzed by use of the Statistical Analysis System (SAS Institute, 1985) . Soil and forage were analyzed

PAGE 65

53 as a split-plot design, with farm as the main plot and season as the subplot. For forage, plant part was considered the subplot. Animal tissues were analyzed as a split-plot design with animal class as the main plot and season as the subplot. Significance level was limited to .05 in all statistical analysis. Since the data were unbalanced, hypothesis testing was based on the Type IV Sum of Squares. Correlation coefficients among minerals were determined. Results and Discussion Soil Analyses Soil analyses as related to season and farm are presented in table 5. Summary of the analysis of variance is shown in appendix B table 24. Soil organic matter was lower (P<.05) in San Jorge farm during the rainy season, but the pH was higher (P<.05) for the same farm during the dry season. Reid and Horvath (1980) indicated that maximum rate of cation absorption occurs at pH 5-7. Soil acidity problems are associated with pH levels lower than 5.5 and the presence of exchangeable aluminum in the soil (Sanchez, 1976). Don Benito had a higher (P<.05) aluminum concentration during the rainy season; this farm had the lowest soil pH (4.8) of the three farms. Among the macroelements, soils were most deficient in magnesium in relation to critical levels. Percentage of samples below the critical level of 30 ppm suggested by Rhue

PAGE 66

54 R i z o 10 < M CO o H D W E-i en < Efl W en > r E B H s y to Bl .. K w H • H —. <* en F H W u < H ffl 2 < >• U « a n c ~ij y H oi <; U) pi. 3 ^ o > c i S 8 Q «* Li LT O e W i n W b « a) c a 0) S 01 D * c c i X D) I B (13 01 Q * • J3 bi c v: 09 _, * U -H •W « AJ > H 01 u 01 % 'u 10 > & lA CP CO d d o o o" d IC oi o OOor--^io o" a-h b. o o E S > — 01 e c 13 e s

PAGE 67

55 and Kidder (1983), was 61 for the rainy season and 76 for the dry season. There were differences among farms with San Jorge having the highest levels (P<.05). Metson (1974) stated that there is some evidence that acidity aggravates a magnesium deficiency condition. In general, San Jorge had higher concentrations (P<.05) during the rainy season for calcium, magnesium, and sodium. On the other hand, San Jorge had the lowest phosphorus level but it was not significant (P>.05). This element was 46% deficient in the rainy season and 39% in the dry season. Many tropical soils are generally reported to be deficient in phosphorus (Volkweiss, 1978). Thirty seven % of the samples in the rainy season and 61% in the dry season were below the critical level of 62 ppm suggested for potassium by Rhue and Kidder (1983) . Sodium presented very low levels in the three farms especially in Don Benito and San Francisco. Forage Analyses Forage macromineral, crude protein and IVOMD concentrations are presented in table 6. Summary of the analysis of variance is presented in appendix B table 25. Means and standard errors of each species in a season, are presented in appendix B tables 35 and 36. Even though the forage samples were separated by species, these could not be included in the statistical model because of the uneven occurrence of some of them which produced a large number of empty cells.

PAGE 68

56 O Eh Q W Eh < H OS CO < o Eh Z W y c c > Q 5 < M Eh O g 0• w oT Q H K < u n g£ w H K s < O 6, K O Q II K Z « O K < O W o 28 o o O O G o o o o o o OX IN tN O o o o o D «7! " — o <"1 o o o C rM 01 01 o c o o fo n n S3 32 gg gg 33 SS o; o c o o do do" dd d Ji h f. S S S S 38 S g g * o o m _ o o o d e' o* o't' -Id o o d d CO o o o o o o o O o o o o o o o o r. «j u v u >"O l"O fct <0 <

PAGE 69

57 Lower limits of sheep requirements (dry matter basis) suggested by the NRC (1985) were used to calculate the percentage of deficient forage samples. Sodium was the most deficient macroelement in forage samples (93-94%) according to the critical level of .09% (NRC, 1985). Potassium was the least deficient (only l%) element in forage samples. This element is often associated with high levels of nitrogen and moisture in lush, cool-season grasses (Boling et al., 1979). Magnesium was 31 and 75% deficient for rainy and dry season, respectively. Phosphorus was only 3% deficient in the rainy season but 62% in the dry season. Underwood (1981) stated that phosphorus deficiency is the most widespread and economically important of all the mineral deficiencies affecting grazing livestock. Thirteen and 22% of forage samples were calcium deficient in the rainy and dry seasons, respectively . The results of this study are in contrast to the results of many researchers in the Latin American tropics who have reported relatively low calcium and extremely deficient phosphorus forage concentrations (McDowell, 1985). Surprisingly, only 1% of the samples in the rainy season and 11% in the dry season were deficient for crude protein, according to the critical value of 7% suggested by Minson and Milford (1967). The high protein concentration found in this study agrees with that of Laredo and Anzola (1986) who determined the crude protein level in temperate grasses similar to the ones used in this research. Sanchez

PAGE 70

(1976) stated that organic matter in the soil supplies most of the nitrogen and half of the phosphorus taken up by unfertilized crops; Sillanpaa (1982) stated that the correlation between the nitrogen content of the plant and total soil nitrogen is relatively good. Magnesium in San Jorge forage samples was higher (P<.05) during the dry season but the deficiency for this farm was as high as 50%. Michael (1962) reported that serum magnesium levels in sheep were not correlated with herbage magnesium content, but Fontenot (1982) stated that a magnesium deficiency in ruminants may result from low magnesium concentrations in feeds or a reduction in biological availability of dietary magnesium. Nitrogen level, stage of maturity, excessive potassium level, form of magnesium, and amount of readily fermentable carbohydrate have been considered as components of magnesium utilization (Rosero et al., 1980). Calcium, potassium, phosphorus, sodium and crude protein concentrations were not different (P>.05) among the three farms. The IVOMD was lower (P<.05) for San Jorge during the rainy season but not during the dry season. The values of IVOMD for the forages in San Jorge were not low (58.7 and 57.6) and those for Don Benito (71.1 and 58.9) and San Francisco (72.6 and 53.8) are considered high, which implies that the animals were receiving good guality forage especially during the rainy season. Since IVOMD values were relatively high it seems that production of forage DM/ha in

PAGE 71

59 the paramo is more of a limitation to animal performance than is forage quality. In a previous study, Laredo et al. (1989) found that the production of DM/ha in Don Benito was 2.8 ton and 2.0 ton for H^. lanatus basyn (imported cultivar) and H. lanatus L (native cultivar) ; in San Francisco the production of DM/ha was 2.8 ton and 0.8 ton, respectively. These values were low for the standards of the grass (Watt, 1978) and even lower when compared to other temperate grasses. There was a marked effect of season on all macroelements studied. Magnesium, potassium, phosphorus and crude protein were lower (P<.05) during the dry season. Although sodium was not different (P>.05) between seasons, concentrations indicated that there was a serious deficiency at all times. Calcium had similar values for both seasons. McDowell et al. (1983) stated that as plants mature, mineral concentration declines due to a natural dilution process and translocation of nutrients to the root system; in most circumstances, magnesium, phosphorus, potassium and sodium decline as the plant matures. Crude protein declined from 18.4% to 12% (p<.05). in vitro organic matter digestibility also declined, from 66.7% to 56.7%, but the diference was not significant (P>.05). Animal Tissue Analyses Blood serum and bone macromineral concentrations as related to season and farm and as related to season and

PAGE 72

60 animal class are presented in tables 7 and 8. Results of analysis of variance are presented in appendix B table 26. Blood serum. No differences were found (P>.05) among farms for serum calcium and phosphorus during both the rainy and dry seasons, and for magnesium during the rainy season. Magnesium had a higher value (P<.05) in farm San Jorge (2.18 mg/dl) than farm San Francisco (1.93 mg/dl) during the dry season. Differences (P<.05) among animal classes were found for magnesium and phosphorus. Lambs had higher (P<.05) concentrations for serum phosphorus than ewes (5.2, 5.6 vs 3.6, 2.5 mg/dl) in the rainy and dry seasons, respectively, and higher (P<.05) than yearlings but only in the dry season (4.0 mg/dl). On the contrary, lambs had lower (P<.05) values for serum magnesium than yearlings in the dry season. There were no differences (P>.05) for serum calcium among the three animal classes. The general tendency was for the dry season to have lower values of calcium, magnesium and phosphorus; nevertheless, phosphorus was the only element statistically affected (P<.05) by season (4.71 vs 3.99 mg/dl). The incidence of calcium deficiency as percentage of samples below critical levels (McDowell et al., 1984) was 3% for the rainy season and 94% for the dry season. The incidence of deficiency for San Jorge, Don Benito and San Francisco during the dry season was 100, 97 and 87%, respectively; the incidence of deficiency for ewes, lambs

PAGE 73

61 2 c w en o Q w o o o o Co f\D o O r, 2 vD o O o a fvm ire «,D ^ _ o o o o o o do CO < CO 2 c O ED O <0 i." f z a u ?-, o o r s c m a i tn o O to »'n o — o ri o o do £0 CO n vi — o o O O O O O o" i>. O lO u* 5 a • H E co S Q 6, O o a J 2 B) < I jj r 5 .P tu 3 X 3 H 3 O * 10 s , w m r c 01 01 ID -w m (I t< u s s "P L*| « n x X fN en w .

PAGE 74

62 s f*l ^ <-• tN H O 0> Of(?> Oi (7> in\o r-jui o(B o en tj>ff> O c *in r»ox rHO \o r-«. K Eh ? »_„„_, *„«,,»,.,,.,« Jl 5 OO (N ~J OO <"! Ifl . > B) | mo i/io ov mo* a\*-> oo oo OO rfi — (N CO OOOOOC Eh Q g " c — « Eh en 1 a z * pi \0 ^ C IC 0*W Ofh a> «J (BCD —1 -" [0 -i rN miN OC 1C u*> *• rfi ul IN OO | o u 3 a OO CO OO 0*h OO CO OO o J z -.0 y m o o "« t o TJ TJ 4J -fi t I c r-_« --^o o en <-> » nr* J 2 Orinto rj— o en r-i v£ m v OO V, TJ <*4 ^ 1 X f-> ic m IN rl . >. TJ o w .5 "« " -H *5 rcn oo oo oo — Mm a oeotjicMin or~oooo PI 4J d * m ^ S -5* S . V D. V r---" 3 in £ in w ^ oi id in r» tNO coo in io •* »» 9 X cj w r-. H ^,H CO ^T H M IN— OO IS c a 3 « oc oo oo oo oo oo oo k $ a" in K 41 O A 4) w-i TJ ffi U1 N W Ul i£ TJ If CN "ff TT If TJ 41 CQ • m \0 c io in p-ioi cm(D • j6 >.>.>.>, -* -.3 c e c c c c c S j I t* TJ U TJ 1TJ lj TJ l* TJ I* TJ l* TJ |c 5 1 IQ ; tn h z 5"Ui 5 ""' S e Q < *j > in co m m j »' 3 5 o .« * • — ui tn o a u U3 rN rt -HO c « c V i0 IB co n41 e tn J z I V V V « < rt U W C CJ — £ rj> uQ ui 4J s — rji c -H C n 1C 41 io « M Q 41 <0 -. | 3 U TJ 41 1* S JJ CO 4i 41 >. 41 4i u **j K -H IB U TJ c ^ -H J O U 3 U) 41 »m H 01 C 3 * TJ — TJ •S. TJ X O 1 3 B io » Oi S in tj r -j D C -~ C >. HJ X Q' S b « I £ X E TJ W Wl U CZ CD -t (J 0. X fO £ UTJtT

PAGE 75

63 and yearlings was 100, 74, and 100%, respectively during the dry season. The high incidence of serum samples below the critical level indicated that the sheep had a very severe calcium deficiency during the dry season. Sheep and cattle have hormonal mechanisms which maintain blood calcium concentrations within narrow limits by adjusting the proportion of dietary calcium absorbed and, when dietary calcium is inadequate, by resorbing calcium from body reserves in the skeleton (Rowlands, 1980). Black et al. (1973) reported that serum calcium concentrations may be directly affected by dietary calcium intake, and Steevens et al. (1971) reported that serum calcium concentrations are affected more by the amounts of phosphorus and magnesium in the diet than by calcium itself. Serum calcium, however, is influenced only by severe deficiency, and calcium dietary levels may be a more adequate criterion in assessing status of calcium (CMN, 1973) . Serum inorganic phosphate was deficient in 51% of samples during the rainy season and of 67% during the dry season according to the level of 4.5 mg/dl suggested as a critical concentration by McDowell et al. (1984). Deficiency of phosphorus for farms San Jorge, Don Benito, and San Francisco was 53, 42 and 58% for the rainy season and 71, 62, and 67% for the dry season, respectively. Deficiency of serum phosphorus for ewes, lambs and yearlings was 84, 38 and 23% for the rainy season and 97, 19 and 72% for the dry season. Plasma inorganic phosphate

PAGE 76

64 concentrations are maintained by absorption of phosphorus from the gut, and there is no specific mechanism for bone phosphorus resorption (Jacobson et al., 1972); positive relationships between dietary phosphorus intake and plasma inorganic phosphate concentrations have been observed (Rowlands, 1980). However, serum or plasma phosphate is not recommended as a practical criterion for assessing phosphorus status in cattle or sheep (CMN, 1973). Overall incidence of samples below the critical level of 2 mg/dl of magnesium suggested by McDowell et al. (1984) was 22% for the rainy season and 51% for the dry season. The deficiency for farms San Jorge, Don Benito and San Francisco was 41, 47 and 61% for the dry season. For ewes, lambs and yearlings the deficiency was 52, 81 and 35% for the dry season, in contrast to calcium, serum magnesium concentration depends mainly on the dietary intake of available magnesium (Rowlands, 1980). Levels below 2 mg/dl in plasma are classed as hypomagnesaemic, but magnesium concentration in blood serum does not fall until there is a severe deficiency (CMN, 1973) . Bone. Bone macromineral concentrations as related to season and farm and as related to season and animal class are presented in tables 7 and 8. Summary of analysis of variance is presented in appendix B table 26. There was no season nor farm effect (P>.05) for any of the bone parameters studied with the exception that ash

PAGE 77

65 during the rainy season had a variation among animal classes. Ewes had higher (P<.05) ash concentrations than yearlings (64.2 vs 61.7%), and lambs lower (P<.05) than both classes at 60.3%. Individual evaluation of samples indicated that ash concentrations below a critical level of 66.8% (McDowell et al., 1984), was 100% during the rainy season and 89% during the dry season. Spongy bone of the axial skeleton are the first to demineralize in periods of negative balance (Little, 1972). Calcium mean concentrations were below the suggested critical level of 24.5% (McDowell et al., 1984). Deficiencies were 100% for the rainy season and 97% for the dry season. This is in agreement with the results observed by other researchers (Knebush et al., 1986, Tejada et al., 1987) who reported that when animals were evaluated for calcium status, using blood serum and bone samples, a consistently higher number of animals were determined to be below the critical levels based on bone analysis when compared to critical levels in serum. Furthermore, this supports the concept that cattle or sheep resist depletion of plasma calcium through mobilization of bone calcium, thus making bone a more accurate indicator of calcium levels in the animal. Mean rib bone phosphorus was 99% deficient in both rainy and dry seasons, according to the critical level of 11.5% (McDowell et al . , 1984). This is in agreement with

PAGE 78

66 the results of other researchers who worked with cattle in the tropics (Tejada et al., 1987, Vargas et al., 1984). Adequate forage phosphorus concentration appears to be reflected more accurately in plasma phosphate than bone, even though many researchers prefer bone phosphorus over serum concentrations to evaluate the phosphorus status of an animal (McDowell et al., 1983). Bone magnesium concentrations were similar among farms and among animal classes. Cohen (1987) suggested that bone magnesium may be useful in assessing magnesium status in grazing livestock. Correlation Coefficients of Minerals Blood serum and bone macromineral correlation coefficients as related to season are presented in appendix B table 28. Correlation coefficients (P<.05, r>|.50|) in serum were calcium-phosphorus (r=.76), calcium-iron (r=.65) and calcium-zinc (r=.77) during the dry season. For bone, the correlations were calcium-ash (r=.79) during the rainy season, calcium-phosphorus (r=.80), and calcium-ash (r=.97) during the dry season. Correlation coefficients between serum and bone minerals as related to season are presented in appendix B table 29. Correlation coefficients between serum and bone macrominerals were observed among serum phosphorus-bone magnesium (r=.84) and serum magnesium-bone phosphorus (r=.85) during the rainy season.

PAGE 79

67 In general, mineral concentrations among animal tissues did not correlate with each other with a correlation coefficient greater or egual to J .50 j at the probability level of .05. This demonstrated the problem of finding significant correlation coefficients between soil, forage and animal tissues (Conrad et al., 1980). Hematological Measurements Hemoglobin and hematocrit concentrations and leucocyte counts as related to season and farm are presented in table 9 . Summary of analysis of variance is in appendix B table 27. San Jorge had higher (P<.05) total leucocyte counts (16,838/ul) than Don Benito (7,605/ul) and San Francisco (8,107/ul) during the rainy season. During the dry season the values were much lower for San Jorge and different only to San Francisco. Total leucocyte counts in San Jorge during the rainy season exceeded the normal limit of range in sheep set by Schalm and Nemi (1986) of 12,000/ul. Differential leucocyte count in the same farm, during the rainy season, showed that the percentage of lymphocytes exceeded the limit of range of 75% (Schalm and Nemi, 1986). There were no differences (P>.05) among animal classes for any of the hematological measurements studied, but lambs in the rainy season exceeded the limit of range for total leucocytes (Schalm and Nemi, 1986) and also exceeded the limit of range of 75% for lymphocytes but were below the

PAGE 80

68 TABLE 9. n™*S°^ N ' HEMAT0CRIT COHCENTRATIOH MD LEUCOCYTE COUNTS AS RELATED TO SEASON AND FAR"!. Range of levels Season San Joroc Don Mean Benito S.E. Overall Mean Variable Mean S.E. C Mean S.E. Hematocrit, \ 27-45 rainy 34.0 0.51 40.8 0.67 40.3 0.84 37.6 0.74 39.5 Hemoglobin, g dl 9-15 rainy 11.4 0.15 13.6 0.22 dry ii. e 0.32 12.7 0.27 13.4 0.30 Leucocytes, / U X 400O-120OC rainy dry 16838 d 7277 d 997 316 7605*: 6671 d 322 369 8107 6 4598 6 285 11458 Neutrophils, % 10-50 rainy dry 12.8 24.4 1.21 2.45 18.6 22.1 1.92 1.98 26.0 31.0 1.97 18.0 Lympnocytes, % 40-75 rainy 84. B 1.43 76.5 2.04 65.0 2.68 73.1 2.23 70.3 Monocytes , % 0-6 rainy 0.72 0.15 1.50 0.21 2.00 0.22 1.34 0.24 0.63 Eosinophils, * 0-10 rainy 1.72 0.48 3.28 0.79 3.5 2.56 0.50 2.66 3.55 2.88 Basophils, % 0-3 rainy 0.00 0.09 0.06 0.03* 0.44 d 0.42 0.11 0.76 0.17 0.23 0.08 Means based on the following number of for Don Benito and 21, 40 (rainy, dry) Standard error of means Means among farms in a row with diffe samples: 36, 36 (rainy, dry) for San Francisco. i Jorge, 32, 29 (rainy, dry) rent superscripts differ
PAGE 81

69 range of 10% for mature neutrophils percentage. These lambs exhibited slight to moderate leucocytosis, defined as WBC values of 13,000 and 20,000/ul (Holman, 1944) and could mean that an infectious process was taking place in the lambs of San Jorge at the time the samples were taken. Summary and Conclusions A study was conducted to determine the macromineral status of three sheep farms located on the paramo region of the Cordillera Oriental in Colombia, and to compare animal classes (pregnant-lactating ewes, lambs, and yearlings) . Soil, forage, blood and rib bone samples were collected to correspond to the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988) . Season affected (P<.05) soil concentrations of calcium and sodium and forage concentrations of magnesium, potassium, phosphorus and crude protein. Soil analyses showed high organic matter (19%) and low pH values (5.0) for the three farms. Magnesium was the most deficient macroelement in both seasons with an overall deficiency of 70%, followed by potassium with 52% of the samples deficient; phosphorus was 46% deficient. Sodium was the most deficient macrolement in forage samples with 93% overall deficiency; phosphorus was deficient only in the dry season (62%) and magnesium had 53% overall deficiency. Crude protein was deficient in 6% of the forage samples.

PAGE 82

70 Blood serum analyses showed an overall phosphorus deficiency in 59% of the samples, and a calcium dry season deficiency of 94%. Bone was 98% deficient in both calcium and phosphorus, with ash percentage being also very deficient (95%). Differences (P<.05) among animal classes were found in serum phosphorus in both seasons (lambs were higher in phosphorus) , in serum magnesium in the dry season (lambs were lower in magnesium) , and bone ash in the rainy season (lambs were lower in ash percentage) . Among soil minerals and the corresponding forage minerals only calcium and magnesium had positive correlation (P<.05, r > |.50|) coefficients for both seasons. In general, macrominerals between animal tissues did not correlate with each other. Based on these analyses it was concluded that macromineral status of sheep on the paramo was deficient and supplementation programs should provide common salt, calcium, phosphorus and magnesium.

PAGE 83

CHAPTER V MINERAL STATUS OF SHEEP IN THE PARAMO REGION OF COLOMBIA. II. MICROELEMENTS Introduction It has been reported (McDowell and Conrad, 1977) that ruminants grazing forages in severely mineral-deficient areas may even be more limited by this condition than either a lack of energy or protein, and that trace element deficiencies or imbalances in soils and forages are responsible for low production and reproduction among grazing livestock (McDowell et al., 1984). As grazing livestock usually do not receive mineral supplementation except for common salt, they must depend almost exclusively upon forages for their requirements. Only rarely, however, can forages completely satisfy all mineral requirements (Miles and McDowell, 1983). It has been shown from a region in the Colombian paramo that forages are low in copper, cobalt and zinc and high in iron and molybdenum and that this imbalance in micronutrients might be the cause for the low production of the sheep in the area (Proyecto Ovino Colombo Britanico, 1979) . 71

PAGE 84

72 The objectives of this study were to evaluate the micromineral status of grazing sheep and to determine the microelement status of soil and forages of three sheep farms as related to the two seasons prevailing in the paramo region of Colombia. Materials and Methods Soil, forage and animal tissue samples were collected from three sheep farms in the Cordillera Oriental of the paramo of Colombia. Sampling periods corresponded to the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988) . A total of 113 composite soil, 131 composite forage, 207 serum and 113 liver biopsy samples were obtained for each of the sampling periods. Each composite soil sample for each farm was obtained from 8-12 samples. A sampling technique described by Bahia (1978) was used. Soil samples were analyzed by standard methods in the Laboratorip Nacional de Suelos of ICA in Bogota for boron, copper, iron, manganese and zinc. Minerals were extracted from the soil samples .with a double acid extracting solution (.025N H 2 S0 4 and .05 N HC1) . Soil mineral concentrations were determined by atomic absorption spectrophotometry (Perkin-Elmer, 1980) . Based on texture, the soil in the upper part (above 3,000 m) of San Jorge is classified as loamy (about 50%) or

PAGE 85

73 silt loam (about 50%); in the lower part (below 3,000 m) about 40% of the soil is clay, 40% a combination of clay loam and sandy clay loam, and the rest is sandy clay, loam, and sandy loam, in Don Benito the predominant soil is loamy, and in San Francisco is either loamy or silt loam. Each composite forage sample was obtained from 20-25 samples of the same forage species predominating and most frequently grazed by sheep in the different areas of the farm. The forage species collected were vernalgrass (A. odoratum) , a native cultivar of velvetgrass (Bi lanatus L) , an imported cultivar of velvetgrass (H. lanatus basyn) , kikuyugrass (P. clandestinum ) , white clover (T. repens ) , tall fescue (F. arundinacea ) , and orchardgrass (D. glomerate . Appendix B table 21 shows the detailed number of soil and forage species; proper identification of paddocks and of forage species is in appendix C. Forage samples were processed and analyzed for mineral content according to methods described by Fick et al. (1979). Each of the farms maintains about 3 sheep/ha/year under a rotational grazing system. There is not a fertilization program in San Jorge; however, some paddocks in the lower part of the farm receive nitrogen (in the form of Urea) at 50 kg/ha/year. In Don Benito and San Francisco pastures are not fertilized; however, some paddocks take advantage of residual fertilization as a consequence of potato cropping. Samples were collected from animals that were divided into three classes: lactating (or pregnant) ewes, lambs (l-

PAGE 86

74 4 months of age) and yearlings (10-14 months of age) . The sheep were Criollo x Romney or Criollo x Corriedale in San Jorge and Criollo x Blackface in Don Benito and San Francisco. Blood plasma and liver biopsy samples were collected from the animals at each farm as described by Fick et al . (1979) and McDowell et al . (1983). Selenium analyses of soil, forages, blood and liver were carried out using a fluorometric technique described by Whetter and Ullrey (1978) . Forage iron, copper, manganese and zinc, blood copper and zinc, and liver copper, iron, manganese and zinc were determined using atomic absorption spectrophotometry using a Perkin-Elmer AAS 5000 (Perkin-Elmer, 1980) . An atomic absorption spectrophotometer equipped with a graphite furnace and Zeeman background corrector (Perkin-Elmer Model 3030) was used to determine forage and liver cobalt and molybdenum. Data were analyzed by use of the Statistical Analysis System (SAS Institute, 1985). Soil and forage were analyzed as a split-plot design, with farm as the main plot and season as the subplot. For forage, plant part was considered as the subplot. Animal tissues were analyzed as a split-plot design with animal class as the main plot and season as the subplot. Significance level was limited to .05 in all statistical analyses. Since the data were unbalanced, hypothesis testing was based on the Type IV Sum

PAGE 87

75 of Squares. Correlation coefficients between minerals were estimated. Results and Discussion Soil Analyses Soil micromineral analyses as related to season and farm are presented in table 10. Summary of analysis of variance is shown in appendix B table 30. Description of paddocks and forage species are in appendix C. Soil iron was higher (P<.05) in Don Benito than San Francisco during the dry season. A similar trend was found during the rainy season. According to the critical level of 2.5 ppm set by Viets and Lindsay (1973) none of the samples for the three farms were deficient in iron. These concentrations were considerably in excess of normal values, and could result in a reduced availability of phosphorus to plants (Lindsay, 1972) . San Francisco farm had a lower (P<.05) soil zinc value than the other two farms during the rainy season. Individual evaluation of samples based on a critical level of 1 ppm when pH is below 6.5 (Rhue and Kidder, 198 3) indicated that none of the samples during the rainy season, and only 13% during the dry season were deficient for this element. San Francisco had a lower (P<.05) soil boron concentration than the other two farms during the dry

PAGE 88

76 m o IB 01 1C f > U -. h ai « O Of ai -m Op D" B = 4 01 U M

PAGE 89

77 season. The percent deficiency of this element, according to the value of 0.4 ppm suggested by Gammon (1976) was 85% for the rainy season and 80% for the dry season. Boron has been established as essential for higher plants and is added frequently to fertilizers for plants with high requirements such as alfalfa (NRC, 1980) . Farm was not a source of variation (P>.05) for copper and manganese during both seasons, for boron and iron during the rainy season and for zinc during the dry season. The only microelement affected (P<.05) by season was zinc which had a lower value during the dry season. Copper, iron and manganese had lower values during the dry season but were not different (P>.05). The percentage of samples below the critical level of .3 ppm for copper (Rhue and Kidder, 1983) was none during the the rainy season and 17% during the dry season. Most of the deficient samples for copper were found in Don Benito, with 57% of the samples deficient during the dry season. For manganese, the percentage of samples below the critical level of 5 ppm (Rhue and Kidder, 1983) was 44% during the rainy season and 59% during the dry season. Among farms the deficiency was San Jorge, 30 and 35; Don Benito 53 and 79; San Francisco, 50 and 70% for rainy and dry seasons respectively. Soil selenium was not included in the statistical analysis but the concentrations for the three farms during the rainy season are presented in table 10. The selenium

PAGE 90

78 concentration of soil reflects that of the parent material (Reid and Horvath, 1980) . Some research has indicated that soil selenium concentrations of less than .5 ppm are found in areas where selenium deficiency in livestock occurs (McDowell et al., 1989). Based on this critical level, all samples were deficient in selenium. Forage Analyses Results of forage micromineral analyses as related to farm and season are presented in table 11. Summary of analysis of variance is shown in appendix B table 31. Forage cobalt was higher (P<.05) in San Jorge in comparison to Don Benito and San Francisco during the rainy season. A similar tendency was observed during the dry season, but was not significant (P>.05). According to the critical level of .1 ppm suggested by the NRC (1985), 79% of the samples were deficient in cobalt during the rainy season and 95% during the dry season. Among farms deficiencies were San Jorge, 54 and 90; Don Benito, 100 and 100; San Francisco 95 and 100% for rainy and dry seasons, respectively. McDowell et al. (1982) reported that, with the exception of phosphorus and copper, cobalt deficiency is the most severe mineral limitation to grazing. livestock in many tropical countries, and sheep are more severely affected than other ruminant species. Manganese concentration was higher (P<.05) in San Francisco in comparison to San Jorge during the dry season. According to the value of 20 ppm (NRC, 1985) only 1% of the

PAGE 91

O m o r~ en 79 Q < o oc OM oo OiA ho Ol ' O O O U1 o 2 < Cm Q s o CO a w CO o £ c > o o o o o O O O O ff(N dp-' o o OO O rsi £• p 5 H? wo -h(n low (Titn trroo co £ no S3 oo wv * N ° o o'o do tn'o VO CO O O o O O O O o \0 "» O O I CO in l» ~ _ ,-; g. w u z c u H s **> 0"i tn CTi o o «r» ; « OO oj ro*o ™c oo w o o d d d -h' d r-' d d d o" t^ n o d a. ~ 4J * 5 £ i 5 o w S CO p < fa en J *C b -O kj -o o O o £ | a I i 1 * i i I ca a. a £ ft * 8 * 8 s e £ £ • fi CJ c

PAGE 92

80 samples were deficient during the rainy season and none during the dry season. All values were in excess of the requirements for sheep. Several studies indicate the ruminant has a high tolerance for manganese (Hansard, 1983) ; mineral imbalances typified by excesses of iron and manganese may interfere with metabolism of other minerals (Lebdosoekojo et al., 1980). The farm means were well below the maximum tolerable level of 1000 ppm (NRC, 1985) . Forage selenium was higher (P<.05) in Don Benito compared with the other two farms during the rainy season. Forty-three % of the samples during the rainy season and 69% during the dry season were deficient according to the .1 ppm value suggested for sheep by NRC (1985). Among farms the deficiencies were: San Jorge, 50 and 70; Don Benito, 25 and 53; San Francisco, 53 and 80% for rainy and dry seasons, respectively. Underwood (1981) considered that for all ordinary grasses and legumes the primary determinant of selenium concentration was the level of available selenium in the soil. Forage molybdenum concentrations were different (P<.05) among farms during the rainy season. Don Benito had a higher value in comparison with the other two .farms. However, none of these values were above the 6 ppm (dry basis) suggested as a toxic limit (McDowell et al., 1984). According to Suttle (1986) only a small increase in molybdenum and sulfur concentrations will cause major reduction in copper availability. He reported that

PAGE 93

81 differences of 3 ppm of molybdenum (from 1 to 4 ppm) and .5 ppm of sulfur (from 2.5 to 3) between two pastures are sufficient to reduce copper availability to one half. Farm was not a source of variation (P>.05) for copper, iron and zinc during both seasons; for cobalt, molybdenum and selenium during the dry season and for manganese during the rainy season. Season was a source of variation (P<.05) for the following microelements: cobalt, copper, manganese and molybdenum; all except manganese had lower (P<.05) concentrations during the dry season. The levels of selenium and zinc were also lower during the dry season, but were not significant (P>.05). On the contrary, iron and manganese were higher during the dry season. The percentage of samples below the critical level of 7 ppm suggested by NRC (1985) for copper was 57% for the rainy season and 94% for the dry season. Among farms the deficiencies were as follows: San Jorge, 36 and 97; Don Benito, 65 and 87; San Francisco, 79 and 95% for rainy and dry seasons, respectively. The results of this study showed that copper: molybdenum ratio was at least 6:1 in San Jorge and San Francisco and 4:1 in Don Benito, and that molybdenum levels were not higher than 4 ppm in any case. Copper concentration was marginal in forages during the rainy season and deficient during the dry season. The animals, in this situation, might respond to appropiate copper supplementation.

PAGE 94

82 Evaluation of samples based on the dietary zinc requirement of 20 ppm (NRC, 1985) indicated deficiencies of 28% during the rainy season and of 45% during the dry season. Among farms, the deficiencies were: San Jorge, 11 and 57; Don Benito, 20 and 13; San Francisco, 63 and 50% for rainy and dry seasons. Ruminants have exhibited signs of zinc deficiency when grazing forage containing 20 to 30 ppm of zinc (Pierson, 1966) . However, a marginal zinc deficiency appears to be a more widespread occurrence (Spears, 1989) . Forage samples evaluated on the levels of 3 ppm of iron (NRC, 1985) showed that only 6% were deficient during the rainy season and none during the dry season. This is in agreement with the zero incidence of iron deficiency found in the soil samples. McDowell (1985) stated that ruminant animals are not likely to suffer from iron deficiency. None of the samples reached the maximum tolerable level of 500 ppm suggested for sheep by the NRC (1985) . Animal Tissue Analyses Blood serum and liver micromineral concentrations as related to season and farm are presented in table 12, and as related to season and animal class are in table 13. Summary of analysis of variance of serum and liver microminerals is shown in appendix B table 32. Blood serum. There were no significant farm by animal class interactions (P>.05) for any of the serum microminerals considering seasons separately, iron,

PAGE 95

83 — CC O >» rin CO »» mm oifl "in Oi" IO •O a 3 c z 01 "ffj o 4 tj 11 n I NO «« Oh l»» m eo in n c CO B — O 1 R 01 O O «r \o O n hr-wnonipr-oi*io *2 cm in — _* ,n -r r-g int\ eo a w 8 3 C U E n in "»m oo f~-o in n-nintNOOnnoOOO O c c ^ CO < en c 1 vvininwr-t-rui n gg — e _, Of a h OO h ffl in -h r-n >»* (Nto »ho in o o id — >• 1 •— -h Oo oo (NfM w i\ r~in i-ieo (N n OO OC -< CO 1 rt 9s c — ? -4 H -1 J8 H It." • — o O O IN (N iD n hOnr^Oon-NOC IO l Z Eh -H K c •U = n E T. a OOOOCO<-»0 niOiOCDOnlNWOOO© rN = * O m OO OO OO OO « ff C O fn -N ro OO OO o" ,5 *" i • " 4 k> S 7 j ^p'TO'^Offir•*0 f N iflin rh e S c * i i 10 IN rin H H f~-o O-h OO OO fN(N (Tin on n ij\ inrN in fh mo o fa « Cy "*>» inffi n in r» co oo OO --• n c -I fn o> O K k >. 01 H = *E H >. h s S "3 B K T CJ 4 -rnf-io-NOrtfi aj COCOODi-lf^ojr'e n e — 1 £ 3 1) OO O-H OC -n fn OO OO OO OO c?*iO Pio ^ 0} "T -4 OO -NO -4 in c 01 01 torro oo coco 0*0 oo Q c tT-JOCCwu-ihtt e* 03 tf> N r> m o r*cr-r-F40'"0 •»a>coiniD--HiahrMMioo cr> SI u — V a > £ V Ofnooooinin minr-cohFNr-noddd o fa TJ < -j* m -H co co a ° s 3 u n o; . >•>•>.>. >•>->.>.>,>.>, — I to £ „ < 3 0) C C C C CCC ccc -j ^f ^> -^ «>-.f:>.5>.=>,^>.=*,^ > , £ £ £ ii £-rt Si; Pi: Pi: ? is m u m u * u * * H TJ T) U TJ H TJ -.TJ U TJ -i TJ H U I. TJ H "0 . TJ 0' a tc s o nm * to .5 o a iJ 2" -« u * « S g JQ 5 — — c c a < H * o o o omicvc'Tj-o' f uco ffi ,-. FN m >. c u 1 — j: -. UN V) U TJ U c (Nl 4 •J -H U U 01 8 S r~\ c c *J TJ --< H3 01 01 A U K § tn_.[i. j3 I e E E a E u -1 u — E. ". _ Z. "_ jl Br D. B B Eli S S « IB — — J3 C a « ij TJ Id c TJ « C 01 H > S'f a c oi J -* wl u n tn u. j • p s a • h. U E N O E w TJ £ tfl U TJ

PAGE 96

K c < M CO C to a CO % o Eh SS K u 2! o B H s g O CO w 8 W H Q < O O Q J Z CQ < a i jo io it orro> CO OO OO — < o OO OO OO OO vO JJ O O CO C-H OO rl — OO OO OO OO CO OO OO INN OO OO OO -H-* OO OO OO OO O rt O O O o TJ II U J 1 (Jt OO CO ft OO OO • <£> OCT" O CO (E 51 Oli P — i UVLfl qT> O OO OO JO OO r»V OO OO O O O O rH iO CTt mo in h i/i r» oo So -i CO OO ft ifi OO oo o en rfirloin no mo \CO O 3t U „tj Tj >i S * c £ 1Ot ~ T ri CO "0 EN C <0 2 to 11 v* o T I>• <0 c > H C a u » 0) a ft tfi u o rrQ 13 9 S8 w ' u 1 0/ — It > a u E . ** C -H i |l 1 3 OUT 1 C 5 u -o 0)
PAGE 97

85 selenium and zinc tended to be higher during the rainy season, and copper higher during the dry season, but none was affected (P>.05) by season. The percentage of serum samples deficient in copper, according to the value of .6 ppm (McDowell et al., 1984) was 18% during the rainy season and 0% during the dry season. Among animal classes, the deficiencies in copper were 19% for ewes, 24% for lambs and 14% for yearlings during the rainy season. Forage copper concentrations were shown to be highly deficient, especially during the dry season (94%) . Serum copper concentrations are often directly affected by dietary copper intakes (Rowlands, 1980) . However, serum copper concentrations do not always reflect dietary copper levels and copper deficiency may occur when serum copper concentrations are high. As discussed by Underwood (1981) , copper deficiency is often caused by molybdenum and sulfur, which interfere with the utilization of copper by the animal, but which may induce either low or high plasma concentrations. It has been observed that low blood copper levels are associated with a reduced microbial activity of the phagocytes in the peripheral blood in sheep and cattle and this is possibly due to a reduction in the intracellular activity of the enzyme superoxide dismutase (Grace, 1988) . Based on a critical level of . 03 ppm for selenium (McDowell et al., 1984), only 15 and 18% of the samples were considered deficient for rainy and dry seasons, respectively. There was no agreement with the percentage

PAGE 98

86 deficiency found in the forage (43 and 69%) for this element. Incidence of serum samples below the critical level of .6 ppm of zinc (McDowell et ml., 1984) was 14 and 27% for rainy and dry seasons. Among animal classes, deficiencies were ewes, 11 and 21; lambs, 5 and 22; yearlings 23 and 3 3% for rainy and dry seasons, respectively. Because signs of zinc deficiency are nonspecific, poor zinc status should be considered in cases of unexplained reproductive problems in the ewe (Apgar and Fitzgerald, 1985) . Serum zinc concentrations are a reasonable criterion to determine the status of the animal, however, values are particularly susceptible to stress of the animal during sampling and can fluctuate rapidly (Underwood, 1981) . Iron deficiency anemias are of the hypochromic microcytic type and it has been emphasized (Underwood, 1981) that an uncomplicated iron deficiency has not been observed in cattle or sheep under normal grazing conditions. Liver. Results of liver biopsy micromineral concentrations as related to farm and season are presented in table 12, and as related to animal class and season are in table 13. Summary of analysis of variance .of liver microminerals is shown in appendix B table 32. There were no significant farm by animal class interactions (P>.05) for any of the liver biopsy microminerals considering seasons separately. Cobalt and molybdenum were higher (P<.05) during the rainy season.

PAGE 99

87 Using the upper limit of the critical value of 75 ppm suggested for copper by McDowell et al. (1984), the percentage of liver samples deficient for this microelement were 72 for the rainy season and 51% for the dry season. However, if we use 25 ppm, which is the lower limit of the range (25-75 ppm) given by McDowell et al . (1984), the incidence of deficiencies would be 3 0% for the rainy season and 20% for the dry season. We can take the upper limit as the marginal level for deficiency and the lower limit as deficient. Among farms, the percentage of marginal copper deficiencies were San Jorge, 82 and 74; Don Benito, 93 and 57; San Francisco, 44 and 23% for rainy and dry seasons, respectively. Among animal classes the percent marginal copper deficiencies were ewes, 80 and 60; lambs, 100 and 33; yearlings, 50 and 49%. Undoubtedly, copper is marginal at best in a high proportion (59%) under the present conditions and special attention must therefore be given to this microelement in the formulation of mineral supplements especially in San Jorge and Don Benito. Although the best criterion of copper status is copper content of liver, blood tests are widely used in practice (CMN, 1973). in this study soil, forage, serum and liver analyses indicated 14, 75, 8 and 59% copper deficiency, respectively. Forage appeared to aid in the diagnosis of copper deficiencies in sheep. Blood did not, but as reported by CMN (1973), if values in liver of yearlings fall

PAGE 100

below about 25 ppm, copper concentrations in blood serum start to decrease. For zinc, the percent liver samples below the critical level of 84 ppm suggested by McDowell et al. (1984) was 42 for the rainy season and 4 5% for the dry season. Among farms, deficiencies were San Jorge, 53 and 74; Don Benito, 13 and 21; San Francisco, 56 and 27% for rainy and dry seasons, respectively. There is no "best" analysis to determine zinc status in the animal (McDowell, 1985). In this study overall zinc deficiency in forages was 36%; on this basis forage appeared to aid in the diagnosis of zinc status in sheep. Liver biopsy samples were not analyzed for selenium during the rainy season. The deficiency of selenium according to the critical value of .25 ppm (McDowell et al.,1984), was 0% for the dry season. This is in disagreement with McDowell et al. (1989) that serum and liver selenium concentrations provide good indicators of dietary selenium status in cattle. The deficiencies for selenium shown in this study were 100, 56, 16 and 0% for soil, forage, serum and liver, respectively. These data are similar to the data of Valdes et al. (1988). It can be concluded that, based on results of liver analyses and of the low percentage of deficient serum samples, selenium status of the sheep appeared to be adequate under present conditions. Forage levels, although marginally deficient, appeared to underestimate the selenium status of the animal.

PAGE 101

89 There were no deficient samples for cobalt during the rainy season, according to the .05 ppm level (McDowell et al., 1984). However, 55% deficiency was found during the dry season. This deficiency appeared to be localized on the San Francisco farm, which had 87% of the samples deficient for cobalt. Among animal classes all three had similar deficiencies (55%) during the dry season. Overall deficiency of this element was 21% in liver and 87% in forage, which did not appear to accurately reflect the concentrations in liver. The deficiency for liver manganese was 6% during the rainy season and 15% during the dry season according to the critical level of 6 ppm suggested by McDowell et al. (1984). The deficiency appeared to be localized only to San Jorge during both seasons. Among animal classes, ewes were 13% deficient, lambs, 31 and yearlings, 6%. Only 1% of the forage samples were deficient in manganese. Liver iron means for the three farms were higher than the proposed critical level of 180 ppm (McDowell et al, 1984). However, 14% of samples of this element were low during the rainy season and 19% during the dry season. Liver iron concentrations suggested that most, of the animals had adequate intake of iron. None of the soil samples, and only 1% of the forage samples were deficient for this micronutr ient . Evaluation of liver samples for molybdenum showed that none of the samples were above the suggested toxic level of

PAGE 102

90 4 ppm (McDowell et al., 1984). Molybdenum has been shown to be an essential component of certain ruminant enzyme systems, but it can be toxic if it blocks copper use in the body (McDowell et al., 1983). Correlation (^ efficients of Minpralc Blood serum and liver micromineral correlation coefficients as related to season are presented in appendix B table 29. Correlation coefficients (P<.05, r> |.050| between serum microelements and liver microelements during the rainy season were as follows: serum copper-liver copper (r=.86); serum copper-liver cobalt (r=-.75); liver cobalt-serum calcium (r=-.70). During the dry season the correlation coefficients were: serum zinc-liver iron (r=.90); serum iron-liver manganese (r=-.91); serum iron-liver cobalt (r=.81); serum selenium-liver selenium (r=.84); liver copperserum magnesium (r=-.80); liver cobalt-serum magnesium (r=.80); liver molybdenum-serum calcium (r— .86). in general, the only positive correlations between serum and liver microelements were found for copper (rainy season), selenium (dry season) and serum zinc-liver iron (dry season) . Summary and Concliisinng A study was conducted to determine the micromineral status of grazing sheep in three farms located on the paramo

PAGE 103

91 region of the Cordillera Oriental in Colombia and to compare animal classes (pregnant-lactating ewes, lambs and yearlings). Soil, forage, blood and liver samples were taken at the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988). Season affected (P<.05) soil zinc and forage cobalt, copper, manganese, and molybdenum concentrations. Soil micromineral concentrations suggesting overall (both rainy and dry seasons) deficiencies were selenium (100%) , boron (83%) , and manganese (54%) . Forage microelements with the highest overall deficiencies were cobalt (87%) and copper (75%) ; copper was 94% deficient during the dry season. None of the values for molybdenum were above the toxic limit of 6 ppm for sheep, and the copper: molybdenum ratio was of at least 4:1. Iron and manganese were in excess of the requirements for sheep but were below the maximum tolerable levels. Blood serum analyses showed only small deficiencies; liver showed overall deficiencies for copper (59%) and zinc (44%) , and for cobalt (55%) during the dry season. No diferences (P>.05) were found among animal classes for any of the plasma and liver microelements in both .seasons. Ewes were more deficient in liver zinc (58%) and liver copper (71%); yearlings were more deficient in liver cobalt; and lambs were about as deficient in liver copper (69%) as ewes. Based primarily on liver analysis it was concluded that copper, zinc and cobalt were the most deficient

PAGE 104

92 microminerals. These elements plus selenium should be present continually in all supplementation programs in the paramo . There was a lack of direct relationship between soil and plant micromineral concentrations. Mineral problems in livestock can be predicted by forage and tissue analyses.

PAGE 105

CHAPTER VI MINERAL CONCENTRATIONS IN LEAVES AND STEMS OF VARIOUS FORAGES OF THE COLOMBIAN PARAMO Introduction It is generally accepted that the nutritive value of a forage, measured as crude protein concentration and organic matter digestibility (IVOMD) is greater in the leaves than in the dead leaves or stems (Pitman and Holt, 1983) . Analysis of total above-ground herbage often leads to an underestimation of the nutritional potential of forages since the animal selectively grazes parts of the plant which have greater protein concentration and higher digestibility (Arnold et al., 1966). Nutrient concentrations in different parts of the plant may vary greatly depending upon forage species, the rate of nutrient absorption and movement of nutrients within the plant (Loneragan, 1975) . Of the total mineral concentrations in soils, only a fraction is taken up by plants (Lindsay, 1972) . However, little relationship has been reported between soil chemistry and mineral composition of native vegetation and farm crops (Reid and Horvath, 1980) . 93

PAGE 106

94 The purpose of this study was to determine the mineral concentration of the leaves and stems in forages of the paramo and to relate soil chemistry and the mineral composition of the same forages. Materials and Methods Soil and forage samples were collected from three sheep farms in the Cordillera Oriental of the paramo of Colombia. Sampling periods corresponded to the end of the rainy season (May-June, 1987) and the middle to end of the dry season (February, 1988) . A total of 113 composite soil and 131 composite forage samples were obtained. Each composite soil sample for each farm came from 8-12 samples. A soil sampling technique described by Bahia (1978) was used. Soil samples were analyzed by standard methods in the Laboratorio Nacional de Suelos of ICA in Bogota for organic matter, pH, aluminum, calcium, magnesium, phosphorus, potassium, sodium, boron, copper, iron, manganese and zinc. Minerals were extracted from the soil samples with a double acid extracting solution (.025N H 2 S0 4 and .05N HC1) . Soil mineral concentrations were determined by atomic absorption spectrophotometry (Perkin-Elmer, 1980) . Based on texture, the soil in the upper part (above 3,000 m) of San Jorge is classified as loamy (about 50%) or silt loam (about 50%); in the lower part (below 3,000 m)

PAGE 107

95 about 40% of the soil is clay, 40% between clay loam and sandy clay loam, and the rest is sandy clay, loam, and sandy loam. In Don Benito the predominant soil is loamy, and in San Francisco is either loamy or silt loam. Each composite forage sample came from 20-25 samples of the same forage species predominating and most frequently grazed by sheep in the different areas of the farm. The forage species collected were vernalgrass (A. odoratum) , a native cultivar of velvetgrass (H. lanatus L) , an imported cultivar of velvetgrass (H. lanatus basyn) , kikuyugrass (P. clandestinum) , white clover (T. repens ) , tall fescue (F. arundinacea) , and orchardgrass (D. glomerata) . Appendix B table 21 shows the detailed number of soil and forage species samples; proper identification of paddocks and of forage species is in appendix C. Samples were collected in plastic bags and kept refrigerated at 5° C until further processing. Samples were then hand separated by species and within each species they were further separated into two fractions; stems and leaves. The fractions were not weighed to get the percent composition of the canopy. In this study, then, only the absolute values for each fraction were obtained. Forage samples were processed and analyzed for mineral content according to methods described by Fick et al. (1979). Forage calcium, potassium, magnesium, sodium, iron, copper, manganese and zinc were determined by atomic absorption spectrophotometry using a Perkin-Elmer AAS 5000 (Perkin-Elmer, 1980) . An atomic absorption

PAGE 108

96 spectrophotometer equipped with a graphite furnace and Zeeman correction (Perkin-Elmer model 3030) was used to determine forage cobalt and molybdenum. Phosphorus was determined by a colorimetric method described by Harris and Popat (1954) and included by Fick et al. (1979). Selenium was carried out using a fluorometric technique described by Whetter and Ullrey (1978) . For nitrogen analysis forage samples were digested using a modification of the aluminum block digestion procedure of Gallaher et al. (1975). IVOMD was performed by a modification of the two-stage technique (Moore and Mott, 1974) . Data were analyzed by the use of the Statistical Analysis System (SAS Institute, 1985) . Soil and forage were analyzed as a split-plot design, with farm as the main plot and season as the subplot. For forage, plant part was considered as the subplot. However, the analysis of the forage data did not use species as a variable. Significance level was limited to .05 in all statistical analysis. Since the data were unbalanced, hypothesis testing was based on the Type IV Sum of Squares. Correlation coefficients between soil and forage minerals were estimated. Results and Discussion Plant Fractio ns-Macrominerals. Crude Protein, and IVOMD Forage macromineral and crude protein concentrations and IVOMD as related to season and plant part are presented

PAGE 109

97 in table 14. Summary of analysis of variance is presented in appendix B table 33. Leaves had higher (P<.05) concentrations of the following nutrients: calcium, magnesium and crude protein during both seasons; phosphorus, potassium and IVOMD only during the dry season. Sodium was the only element with no difference (P>.05) between the two fractions during both seasons. The overall means showed that leaves were higher (P<.05) than stems for calcium, phosphorus, magnesium, potassium, crude protein and IVOMD. Within stems all nutrients had lower values during the dry season and within leaves all nutrients, except calcium, had lower values during the dry season. The overall percentage of leaf calcium was more than twice that of stem, and the percentage of deficiencies were more drastic in stems (83 and 94%) than in leaves (18 and 8%) for rainy and dry seasons, respectively. Working with temperate grasses in Colombia, Laredo and Anzola (1986) reported a value of .31% in leaves and .12% in stems, values which agree with the present study. Although significant (p<.05), the value of phosphorus in the leaves (.21%) was not much higher than in the stems (.19%); the percent deficiencies for the stems were 26 and 78% and for the leaves, 6 and 48% for rainy and dry seasons, respectively. Magnesium was found to be a highly deficient element in this study. Percentage of magnesium deficiencies were 78 and 94% for the stems and 23 and 59% for the leaves

PAGE 110

CO < 98 7O Eh Z K O O U Q -< T, V. O H > CO H < CO o K T. C < D iJ K P O c §s H O T. V. h <: t w C 0": U O " c H K < < O W Cm B. 3 ' £ C X C o o o o o o E > E > E > £ > E> E > E > 10 CO ("H g\ OO OO OO OO OO OO to o o o o OO OO OO OO OO OO O O O O HO rhH OO f^ CO l/l ^ OO « OO " ^ i-> -» e c e c . c .„ >, .H >. H >. •« > -H >. UT3 l--a U 73 HO IHT3 o o o c3 5 10 w C = 1 II II » 5 § £ W Z « J3 ,*••

PAGE 111

99 during rainy and dry seasons, respectively. These data disagree with that of Montalvo et al. (1987) who found lower magnesium concentrations in leaves than stems. For practical purposes, animals grazing either one of the two fractions would not meet their requirements for magnesium. For potassium, most of the samples had higher concentrations than the reported requirements; leaves had higher (P<.05) potassium concentrations than stems, but the percent deficiency in the stems was quite low (13%). There was no difference in sodium concentrations for the two fractions; both parts were highly deficient for this element during both seasons. Although there was a difference (P<.05) in the concentration of crude protein between the two fractions, stems provided higher protein than the requirements during the rainy season but failed to do so during the dry season. Percent deficiency for stems was 2 6 and 67 and for leaves 8 and 9 during the rainy and dry seasons, respectively. A similar trend occurred with the IVOMD values between the two fractions. This could mean that the forages under this study are of good quality and provide adequate protein for the most of the year. Plant Fractions-Microminerals Forage micromineral concentrations as related to season and plant part are presented in table 15. Summary of analysis of variance is presented in appendix B table 34.

PAGE 112

00 00 00 jjjjLI WJJ U II wop 100 io 00 d . O* CO OO rtO 00 00 00 00 00 rco 00 -no 00 Eh 2 « U z • ~ u 0: 1-1 a H s > H « £* a c ~ « U Eh H (K a < Pw O Eh gi ^ 6j c OO O -H o o 00 00 00 o o o" o o o o U V U T3 U -O • -D Ij "C T3 I Eh s S3 -0 K ^

PAGE 113

101 Leaves had higher (P<.05) concentrations of molybdenum during the rainy season, higher concentrations of iron, and manganese during the dry season, but were lower (P<.05) than stems in zinc concentration during the rainy season. The overall means showed that leaves were only higher (P<.05) than stems in iron and molybdenum concentrations. Within stems, iron, manganese, and molybdenum had higher values during the dry season; copper, zinc, cobalt, and selenium were higher during the rainy season. Montalvo et al . (1987) and Laredo and Anzola (1986) also found higher concentrations of iron in leaves than stems. Percent deficiency of this element, according to the 20 ppm critical value (NRC, 1985) , was only 4% for leaves and 10% for stems. Concentration of iron in forages suggested that sheep in the paramo do not need supplemental iron in the diet. Molybdenum was higher (P<.05) in leaves than stems, but none of the individual values were higher than the 6 ppm suggested by NRC (1985) as the toxic limit. Cobalt was equally highly deficient comparing the two fractions. According to the .1 ppm level (NRC, 1985), leaves were 85% deficient and stems 92% deficient. Along with copper, these elements proved to be the most deficient in the forages of this study, and both were potentially limiting grazing sheep productivity in the area.

PAGE 114

102 For copper the deficiencies were 65% in the leaves and 82% in the stems. According to these results, copper should be provided to sheep in the mineral mix. Laredo and Martinez (1984) reported that both zinc and copper were the microelements most affecting production and reproduction of beef cattle in Colombia. Zinc presented 46% deficiency in stems and 42% in leaves, according to the 30 ppm value (NRC, 1985). Montalvo et al. (1987) found much greater zinc levels in the stems than in the leaves. Selenium was highly deficient especially in stems during the dry season (92% deficiency) using the critical level of .1 ppm (NRC, 1985). In general, stems were 77% selenium deficient and leaves 65%. Manganese was consistently higher in both fractions than the recommended value of 40 ppm (NRC, 1985) . It is not advisable to supplement sheep with manganese as it can interfere with the utilization of other minerals. Forage Species Minerals, crude protein and IVOMD concentrations of forage species collected during the rainy and dry seasons, presented with overall species deficiencies are in tables 16 (macrominerals) and 17 (microminerals) ; presented with individual species deficiencies are in appendix B tables 35 (macrominerals) and 36 (microminerals) . Concentrations tended to differ, but the differences were not statistically analyzed, because of the uneven occurrence of some of the species which produced a large number of empty cells.

PAGE 115

103 OS Q « a, m w SB C W u Si o u Q s: 5 K H O « P w Q a u SB § o E O < fc. CO K J I O O O O O O O O (N in O O O O ' O OO OO COn o> ) O OO OO Oh (N 00 m r-j 00 oo do " t o o o o oo oo oo oo do h c rin eoio fvfN co oo oo oo do dd o'o fNr< O\o ho oo f\ h m. oo i-i-h oo do do dt °. c . T"! °.° °° °° °> & oo oo oo oo oo o'o --i -i iO _l _ CO
PAGE 116

& 3 Ji ul \£ O -" O 104 iO IN O o ^ * O O O O "> f > o ort\o H o c co c vO O O to w H u w p W X B § oro o co c ) •£ -ho o o cn kO tin O O O O T O O « 01 O O O O i rO O O O W 8 > O O O O f.O O O o o a o pu o o o o on o o ifN o o « V 01 d .-< O 01 C O O O O n l ..1 a u S OO OO 01 O OOD oo oo £ TJ [i > >. >. hi >. : = c c c c 4 >. -H >. -H >. -rt >, -H > -H >, 3 ij miflu fl ^ AM « O O O i£ o o V V V A V V | & | & | | ft ft a a a a & a s 6 i i i £ i C 5 <0 « .. m ^ T3 — e -h c B E X O U) Z

PAGE 117

105 Because it may be of interest, means and standard errors of each species in a season, are presented. There is a variety of forage species which have colonized the paramo, and for the purpose of this study they are considered native to the paramo. Among them, the most important for grazing sheep are Holcus lanatus L (velvetgrass) , Anthoxanthum odoratum (vernalgrass) , Lachemilla orbiculata (plegadera) , and Tri folium repens (white clover) (Ferguson et al., 1987). Pennisetum clandestinum (kikuyugrass) is an aggresive species which grows well up to 2900-3000 m. Samples of this grass were only obtained in the lower part of the farm San Jorge. The reason two ecotypes of {L. lanatus were included in this study was because of canopy structure differences between the two. The native ecotype, H± lanatus L, produces many erect, unbranched stems with few leaves; matures very early and is a prolific seed producer. In contrast the imported ecotype, H lanatus basyn, forms a denser canopy with more leaves, and is not a seed producer during a considerable time (Mack and Pastrana, 1987). Among the introduced grasses of value to the paramo are Dactyl is glomerata, Festuca arundinacea and Lolium perenne . They have the disadvantage of being easily overcome by the native species. Mean calcium concentrations ranged from .24% in H. lanatus basyn to 1.53% in T. repens . According to the value of .2% as a critical level for sheep (NRC, 1985), H^. lanatus

PAGE 118

106 basyn was 27% deficient for calcium considering both seasons. For potassium, none of the species (except A. odoratum, with 2% deficiency) were deficient, according to the .5% level (NRC, 1985). The lowest mean magnesium concentrations were found in A. odoratum (.09%), H. lanatus basyn (.12%) and D. glomerata (.12%) and the highest was found in T. repens The percent deficiency was 77, 62, 67 and 0%, respectively. All species were highly deficient in sodium, according to the level of .09% (NRC, 1985). P. clandestinum was 100% deficient and T. repens 40%. Mean phosphorus concentrations ranged from .15% in A. odoratum to .41% in D. glomerata and F. arundinacea . The same species showed a deficiency of 56, and 0%, according to the .16% value for phosphorus suggested by NRC (1985). For crude protein and IVOMD, species were different as related to season. All species had higher values of protein and IVOMD during the rainy season. The overall mean protein concentration ranged from 11.4% for A. odoratum to 27.9% for T. repens , and IVOMD ranged from 52.3% for P. clandestinum to 73.7% for D^ glomerata. Among the microelements, high deficiency percentages were found for copper, cobalt, selenium and zinc. Mean copper concentration ranged from 3.2 4 ppm in A. odoratum to 9.27 ppm in F. arundinacea . Percent deficiencies were 93% for A., odoratum and 20% for T. repens . according to the critical value of 7 ppm (NRC, 1985) .

PAGE 119

107 H. lanatus basyn and D. qlomerata had the lowest mean values of cobalt (.03 ppm) and P. clandestinum (.14 ppm) , the highest. The percent cobalt deficiency in P. clandestinum was 33%, and A. odoratum was 98% deficient. For selenium, the values ranged from .07 ppm (A. odoratum and £• arundinacea) to .16 ppm (P. clandestinum). According to the critical concentration of .1 ppm (NRC, 1985), A. odoratum was 72% deficient for this element. H. lanatus L had a zinc mean value of 19.4 ppm which is below the critical value of 20 ppm suggested by NRC (1985). Percent deficiency for the element was 46%. None of the species had iron or manganese mean values below the critical levels recommended by NRC (1985). However, P. clandestinum presented 20% deficiency for iron and 7% for manganese. Excluding A. qlomerata and F. arundinacea from overall comparisons, A. odoratum had the lowest values for potassium, magnesium, phosphorus, copper, selenium, and crude protein; T. repens had the highest values for calcium, potassium, magnesium, sodium, phosphorus, copper, molybdenum, zinc, crude protein and IVOMD. Comparing the two ecotypes of H. lanatus . H. lanatus basyn was higher than the native grass in potassium (2.62 vs 2.13%), sodium (0.4 vs 0.3%), phosphorus (.24 vs .21%), copper (6.23 vs 4.49 ppm), iron (116 vs 103 ppm), manganese (268 vs 232 ppm), molybdenum (1.09 vs .64 ppm), selenium (.12 vs .10 ppm), zinc (22.8 vs 19.4 ppm), protein (19.5 vs 14.6 %) and IVOMD (68.7 vs 65.5 %) . However, the

PAGE 120

108 differences were not extreme enough to claim H. lanatus basyn (imported) to be a better ecotype. If production of DM/ha is higher in H. lanatus basyn, as reported by Laredo et al. (1989), then, this ecotype is of much greater value than the native ecotype for grazing sheep on the paramo. With the exception of provided salt, sheep depend entirely on forages to meet their mineral requirements. However, as shown in this study, the predominant forage species of the paramo do not meet mineral requirements; therefore, mineral supplementation, apart from common salt, is needed in the area. Soil-Plant Relationship Results of correlation coefficients between soil and forage minerals as related to season are presented in table 18. Mineral concentration of a plant depends to some extent on the mineral concentration of the soil. However, soil properties and conditions markedly affect mineral uptake and utilization by plants (Wilkinson, 1972). Correlation coefficients (p<.05, r> |.50|) between soil minerals and the corresponding forage minerals, during the rainy season, were as follows: calcium (r=.81), magnesium (r=.89), potassium (r=.71), zinc (r=.69). During the dry season, the correlation coefficients were calcium (r=.61), and magnesium (r=.58). Only calcium and magnesium had positive correlation coefficients (P<.05, r> |.50|) for both seasons of the 12 mineral elements evaluated. These results showed that only a few minerals have a high correlation

PAGE 121

109 I i£ O — Of<1 r o SS B ™ 9 : i£ -* C* III Di O U). f— CO «n — q rj , w ^ til *~i mo vof < H o w w £ Ed EC 0. E-i w • H 2 O' O h en In < En Ed W v. o u o z o a H w Eh Eh K O W u < J m < on omo mv £ ® >£C Oir\ voo O-I n° £2 ^ ™ "^ !S §8 S; §s ss ss ss vC ^ O-i o ?RSSSS? + sg2SSS2S3S3f ; . a 6 £ c F | .0 W "0 -"I'D l_.-c I-.-D i«>i J -D >t3 1>0 1m -o 8 & & h = o £ o a. c 1« tr< o U o a.

PAGE 122

110 between soil and forage values. Therefore, soil analyses are not of great importance when assessing mineral status of sheep in the paramo. Correlation coefficients between soil organic matter and forage nutrients for the rainy season were magnesium (r=-.76), zinc (r=-.63), and cobalt (r=-.71) . For the dry season, correlation coefficients for soil organic matter were magnesium (r=-.74), iron (r=.64), and cobalt (z— .59). Correlation coefficients between soil pH and forage nutrients for the rainy season were calcium (r=.56), magnesium (r=.58), manganese (r=-.67), and cobalt (r=.54). For the dry season, correlation coefficients were magnesium (r=.61), iron (r=-.53), and manganese (r=-.68). Hodgson (1963) reported the effect of pH on mineral absorption by plants, which could be modified by plant species and by the amount and form of the element in the soil. Summary and Conclusions A study was conducted to determine the mineral composition of two plant fractions (leaves and stems) of several forage species in three sheep farms of the paramo in Colombia. The mineral composition of the forage species, and the relationship between soil chemistry and the mineral composition of the forages were also determined. Sampling periods corresponded to the end of the rainy season (MayJune, 1987) and the middle end of the dry season (February,

PAGE 123

Ill 1988). Leaves were higher (P<.05) than steins in the following: calcium, phosphorus, magnesium, potassium, iron molybdenum, crude protein, and IVOMD. Overall percentage of deficiencies was as follows: calcium (88, 13), phosphorus (50, 27), magnesium (85, 41), potassium (13, 3), sodium (94, 96), iron (10, 4), copper (82, 65), manganese (7, 0), zinc (46, 42), cobalt (92, 85), selenium (77, 65), crude protein (45, 8%) for stems and leaves, respectively. Based on analyses, mineral, crude protein and IVOMD concentrations of forage species, tended to differ. Soilforage correlation coefficients of the same mineral for the rainy season were calcium (r=.81), magnesium (r=.89), potassium (r=.71), zinc (r=.69). For the dry season, correlations were calcium (r=.61) and magnesium (r=.58). Results indicated low correlation coefficients between soil and forage minerals, and that they are not of great value in assessing the mineral status of grazing sheep in the paramo.

PAGE 124

CHAPTER VII SUMMARY AND CONCLUSIONS A study was conducted in three farms of the Cordillera Oriental on the paramo region of Colombia to determine the mineral status of grazing sheep by evaluating mineral concentrations in soil, plant, and animal tissues. Sampling collections corresponded to the end of the rainy season (May-June, 1987) and the middle end of the dry season (February, 1988) . Samples were collected from animals that were divided into three classes: lactating (or pregnant) ewes, lambs (1-4 months of age), and yearlings (10-14 months). A total of 113 composite soil, 131 composite forage, 207 blood serum, 192 whole blood, 148 rib bone biopsy and 113 liver biopsy samples were obtained for each of the sampling periods. Soil samples were analyzed for organic matter, pH, aluminum, calcium, phosphorus, magnesium, potassium, sodium, iron, copper, manganese, zinc, selenium and boron. Forage samples were analyzed for the same minerals as in soil (except aluminum and boron) in addition to cobalt, molybdenum, crude protein and IVOMD. Blood samples were analyzed for calcium, phosphorus, magnesium, copper, zinc, selenium and iron. Bone was analyzed for calcium, 112

PAGE 125

113 phosphorus, magnesium, and ash concentration. Liver was analyzed for iron, copper, manganese, zinc, cobalt, molybdenum and selenium. Whole blood samples were analyzed for hematocrit, hemoglobin, total leucocytes, neutrophils, lymphocytes, monocytes, eosinophils and basophils. Season affected (P<.05) concentrations of soil calcium, which were higher in the dry season, and sodium and zinc, which had higher values during the rainy season. Overall incidence of forage deficient samples in decreasing order was as follows: selenium (100%), boron (83%), magnesium (70%), manganese (54%), potassium (52%), phosphorus (46%), calcium (24%), copper (14%), zinc (12%), and iron (7%). The three farms were characterized by their high organic matter (19%) and low pH value (5.0). Season affected (P<.05) concentrations of forage manganese, which had higher values during the dry season, and of potassium, magnesium, phosphorus, cobalt, copper, molybdenum and crude protein which had lower values during the dry season. Overall incidence of deficient forage samples in decreasing order was as follows: sodium (93%), cobalt (87%), copper (75%), selenium (56%), magnesium (53%), zinc (36%), phosphorus (32%), calcium (17%), and crude protein (6%). None of the values for molybdenum were above the toxic limit of 6 ppm for sheep, and the copper .-molybdenum ratio was of at least 4:1. Iron and manganese were in excess of the requirements but were below the maximum tolerable levels for sheep.

PAGE 126

114 Plant fraction was a source of variation for forage minerals. Leaves had higher (P<.05) concentrations for calcium, phosphorus, magnesium, potassium, iron, molydenum and crude protein. Based on analyses, mineral, crude protein and IVOMD concentrations among forage species tended to differ. A high percentage of species were deficient in magnesium, sodium, cobalt, copper, selenium and zinc for both seasons and in phosphorus in the dry season. Among soil minerals and the corresponding forage minerals, only calcium and magnesium had positive correlation (P<.05, r> |.50|) coefficients for both seasons of the 12 mineral elements evaluated. Therefore, soil analyses are not of great importance when assessing mineral status of sheep in the paramo. Season only had effect (P<.05) on the following animal tissues: serum phosphorus, and liver cobalt and molybdenum, which had lower concentrations during the dry season. Serum analyses showed an overall phosphorus deficiency in 59% of the samples, magnesium 38%, zinc 21%, selenium 16%, and copper 8%; calcium was 94% deficient during the dry season. Bone was 98% deficient in both calcium and phosphorus, with ash percentage being also very deficient (95%XLiver showed an overall copper deficiency of 59%, zinc 44%, and cobalt 55%, and selenium 41% during the dry season. Differences (P<.05) among animal classes were found in serum phosphorus in both seasons (lambs were higher in phosphorus) , lambs were lower in serum magnesium during the

PAGE 127

115 dry season, and bone ash during the rainy season. No differences (P>.05) were found among animal classes for any of the serum and liver microelements in both seasons. Ewes were more deficient in liver zinc (58%) and liver copper (71%) ; yearlings were more deficient in liver cobalt (40%) ; lambs were as deficient in liver copper (69%) as ewes. In general, mineral elements among animal tissues did not correlate with each other. Considering only the minerals which were analyzed in the samples it can be concluded that the minerals most likely limiting sheep production in the paramo were calcium, phosphorus, magnesium, copper, zinc, and cobalt. Supplementation programs for sheep in the paramo should provide common salt, calcium, phosphorus, magnesium, copper, zinc, cobalt and selenium. ,•

PAGE 128

APPENDIX A FIGURES

PAGE 129

E] San Jorge Don Benito UJ San Franciscc FIGURE 1. COLOMBIA, GEOGRAPHICAL LOCATION OF THE THREE SHEEP FARMS SURVEYED. 117

PAGE 130

118 PiSOS TERMlCOS NlEVES PEHPETUA5 UNO < 12°C [U Cordillera Occidental [2] Cordillera Central Q] Cordillera Oriental FIGURE 2. THE PARAMO REGION IN COLOMBIA.

PAGE 131

::' APPENDIX B TABLES

PAGE 132

TABLE 19. DESCRIPTION OF FARMS Description San Jorge Don Benito San Francisco Name of owner: Location of farm : Departamento Municipio Distance from Bogota (km) Land use Total area (ha) Natural pasture (ha) Improved pasture (ha) Agronomic crops (ha) Forest-shrubs (ha) Non-usable land (ha) Pastures Name of predominant forages First ICA Caja Agraria Caja Agraria Cundinamarca Cundinamarca Boyaca Soacha. Zipaquira Ventaquemada 30 50 110 800 836 1100 445 320 480 60 80 90 80 70 110 110 216 260 105 150 160 Second Third p Grazing system Stocking rate (animals/ha) Sheep Principal breeds Total sheep Rams Ewes Lambs Yearlings Lambing (%) Mortality (%) Breeding season Lambing season Weaning age (months) Age female bred (months) Holcus lanatus A.odoratum clandestinum rotational 3.4 Criollo Romney 1710 85 605 526 494 87 6 Aug-Sept Jan-Feb 4 18 Holcus Holcus lanatus lanatus A.odoratum A.odoratum T.repens P. clandestinum rotational rotational 2.2 3.6 Blackface Criollo 872 24 320 272 256 85 6 NovDec April-May 4 18 Blackface Romney 2045 96 720 634 595 88 6 Dec-Jan May-June 4 18 121

PAGE 133

TABLE 19. — CONTINUED 122 Description 8 San Jorge Don Benito San Francisco Parasite control Internal External Mineral supplements ** every month 3 times/yr 4 times/yr once/yr once/yr once/yr yes yes yes Some data were derived from approximation in cases where exact data were not available. •ta ineral supplementation was not a continuous practice in any case.

PAGE 134

123 TABLE 20. CLIMATE AND AVERAGE MONTHLY RAINFALL (mm) Farm Description San Jorge Don Benito San Francisco Type Average temperature (°C) Average rainfall (mm) Rainfall, 1987 (mm) January February March April May June July August September October November December Dry forest Wet forest Lower montane Montane 10 9 640 1300 13.8 21.3 25.3 97.4 35.4 52.0 73.7 100.7 120.0 217.2 13.2 89.4 77.0 125.2 43.5 78.4 42.7 96.1 93.0 220.7 43.7 87.8 21.7 50.5 Wet forest Montane 9 1300 January, 1988 31.8 16.5

PAGE 135

124 TABLE 21. DETAILED NUMBER OF SOIL AND FORAGE COMPOSITE SAMPLES Paddock # Rainy season ( 19871 Soil Forage Species 8 Rainy season (1987) Soil Forage Species San Jorae 1 2 3 4 5 6 7 8 Totals Don Benito 9 10 11 12 13 14 15 Totals San Francisco 28 28 1 2,3 2,3,4,5 2,3,4,6 1 4 4 4 4 2 1 1 20 4 4 8 6 3 2 2 29 4 4 4 4 4 7 2 7 2 1,4,6,7 1 2,3 1,2,3,4 1,2,3,6 1,2 1,2 1,2 1,2 6,7 1 1,2

PAGE 136

TABLE 22. COMPOSITION OF THE MINERAL MIXTURES 3 125 Compound Element San Joroe Common salt 50.0 Steamed bonemeal 49.7 Cupric sulfate .25 Potassium iodide .005 Cobalt chloride .003 Sodium chloride Calcium Phosphorus Copper Cobalt 50.0% 14.4% 6.0% .06% 7.0 ppm Don Benito and San Francisco Common salt Dicalcium phosphate Zinc sulfate Flowers of sulfur Cupric sulfate Potassium iodide Cobalt chloride Sodium selenite 62.0 Sodium chloride 33.0 Calcium 1.7 Phosphorus 1.0 Sulfur .6 Copper .015 Zinc .004 Iodine .003 Selenium Cobalt 62 .0% 10 .0% 6 .0% 1 0% 15% 5% 00 ppm 14 ppm 10 ppn Calculated consumption of mineral mixture: 12 g/head/day in San Jorge and 10 g/head/day in Don Benito and San Francisco. Tlineral supplementation was available during breeding and lactation and occasionally during the rest of the year. c Mineral supplementation was available during most of the year.

PAGE 137

126 TABLE 23. DETAILED NUMBER OF TISSUE SAMPLES Animal class Rainv season (1987 Bone Liver Dry (19881 Serum Blood 3 Serum Blood Bone Liver San Jorge Ewes Lambs Yearlings 12 12 12 12 13 10 11 7 8 8 3 6 10 12 12 11 13 12 12 12 9 8 6 9 Totals 36 35 26 17 34 36 33 23 Don Benito Ewes Lambs Yearlings 13 9 11 12 10 10 11 7 4 8 4 3 6 6 22 4 5 24 15 14 Totals 33 32 22 15 34 33 15 14 San Frsnniqrn 10 10 12 13 9 9 17 9 20 11 4 21 8 19 Ewes Lambs Yearlings 12 12 8 18 Totals 24 20 25 18 46 36 27 26

PAGE 138

TABLE 24. SUMMARY OF ANALYSES OF VARIANCE OF SOIL pH, AND MACROMINERALS— MEAN SQUARES BY 127 ORGANIC MATTER, SEASON Response Farm 3 Paddock/ farm Residual Rainv season Organic matter. 1495.51** 140.08 P H .6272* .2316 . Aluminum 842202.17* 185294.14 Calcium 2571258.49* 504613.84 Magnesium 193431.01* ,. 31442.20 Phosphorus 7774.58 3168.51 Potassium 11364.49 4631.40 Sodium 5769.74** 360.06 df* 2 12 Drv season Organic matter 277.63 140.92 P H 1.6800** .1811 Aluminum 323635.24* 108049.92 Calcium 1357359.20 646938.00 Magnesium 50091.30 21191.18 Phosphorus 706.94 699.91 Potassium 12400.31* 3912.66 Sodium 538.40* 174.21 df 2 13 7.05 .0090 4250.61 10424.34 784.87 177.36 742.66 20.61 44 6.1174 .0329 15913.57 41548.64 1483.12 82.14 349.19 52.48 38 Test farm using paddock/farm as an error term. ""Represents degrees of freedom. P<0.10. *P<0.05. **P<0.01.

PAGE 139

128 TABLE 25. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE CRUDE PROTEIN IVOMD AND MACROMINERALS— MEAN SQUARES BY SEASON Response Farm Calcium Magnesium Phosphorus Potassium Sodium Crude protein IVOMD df b Calcium Magnesium Phosphorus Potassium Sodium Crude protein IVOMD df .2381 .0261* .0472* 5 .9751* .0011 80 .3744* 1236 .18** 2 0428 0164* 0112 1 4409 0003 31. 7166 105. 9609 Paddock/ farm Rainy season .1083 .0090 .0167 1 .7494 .0007 28 .4002 77 .0316 12 Drv season 0385 0040 0089 9772 0026 63. 1011 124. 2633 13 Residual .0840 .0029 .0020 .0009 19.9156 47.5852 52 .0392 .0012 .0011 .1618 .0010 9.7055 28.4418 49 "Test farm using paddock/ farm as an error term. Represents degrees of freedom. *P<0.10. *P<0.05. **P<0.01.

PAGE 140

129 TABLE 26. SUMMARY OF ANALYSES OF VARIANCE OF SERUM AND BONE MACROMINERALS— MEAN SQUARES BY SEASON Response Serum Farm" Class" Farm x class Rainv season Residual Calcium Magnesium Phosphorus Calcium Magnesium Phosphorus .9008 .0212 1.1413 2 4.5138 .9806* 4.6465 6.3642* .2455 37.6635** 2 1.0086 .2165 1.4961 3 Dry season 13.1692 1.1426* 58.8009** 4.7069 .1152 2.0116 Bone Rainv season .5271 .0821 1.7711 85 .5134 .0790 .9489 Ash Calcium Magnesium Phosphorus Ash Calcium Magnesium Phosphorus df 5.9424 2.1997 .0026 6.3506 91.1334 34.7895* .0195* 12.2519 2 79.9057** 16.3216 .0229* 42.3664* 1.5355 3.8251 .0030 8.3914 Dry season 268.2370 40.1908* .0041 9.4867 2 49.5250 2.5943 .0015 4.0728 2 3.7976 .9316 .0028 2.8385 8.4698 6.7941 .0045 2.0671 70 Test farm and class using farm * class as an error term. Represents degrees of freedom. P<0.10. *P<0.05. **P<0.01.

PAGE 141

130 TABLE 27. SUMMARY OF ANALYSIS OF VARIANCE OF WHOLE BLOOD VARIABLES — MEAN SQUARES BY SEASON Response Hematocrit Hemoglobin Leucocytes Neutrophils Lymphocytes Monocytes Basophils df 6 Hematocrit Hemoglobin Leucocytes Neutrophils Lymphocytes Monocytes Basophils df Farm 8 . 315.9647* 48.1726* 575884482** 730.4968 1636.91* 9.4986 .1138 261.9621* 30.6456 58276064* 1729.55 540.2386 .1822 2.3330 2 Class" Farm x class Rainv season 2.5048 1.3066 20004183 382.5963 480.6160 .7951 .0695 2 53.9255 4.9817 16111240 186.2268 221.5605 4.3139 .0985 3 Dry season 57.3294 8.7337 1993866 1048.91 317.2786 1.7832 .3309 2 60.2374 10.4269 4251604 1024.33 1148.18 2.6948 .7082 4 Residual 9.5438 1.0322 16108862 67.7747 75.1212 2.9106 .0524 81 16.6150 2.5663 2798545 526.56 197.5488 197.5488 .4751 96 "Test farm and class using farm * class as an error term. Represents degrees of freedom. *P<0.10. *P<0.05. **P<0.01.

PAGE 142

131 TABLE 28. BLOOD SERUM AND BONE MACROMINERAL CORRELATION COEFFICIENTS AS RELATED TO SEASON 8 Serum Bone Variable Ca Mg P Ca Mg P Ash Rainv season Serum Calcium i.ooo Magnesium .263 1.000 Phosphorus .673* -.137 1.000 Copper .493 .452 -.175 ., Iron -.176 -.173 -.086 Selenium -.448 .112 .020 Zinc -.227 -.156 -.557 Drv season Calcium 1.000 Magnesium -.486 1.000 Phosphorus .765* -.407 1.000 Copper -.414 -.011 -.519* Iron .659* -.753* -.307 Selenium -.284 -.367 -.077 Zinc .778* -.583* .566* Rainv season Bone Calcium 1.000 Magnesium -.674* 1.000 Phosphorus .620* -.465 1.000 Ash .796* Drv season -.939** .486 1.000 Calcium 1.000 Magnesium .402 1.000 Phosphorus .805* .665* 1.000 Ash tz. ; — r~. .977** .315 .722* 1.000 Correlation coefficients based on 8 , 9 (rainy, dry) means for serum; and 8, 7 (rainy, dry) for bone. P<0.10. *P<0.05. **P<0.01.

PAGE 143

132 TABLE 29. CORRELATION COEFFICIENTS BETWEEN BLOOD AND LIVER AND BONE MINERALS AS RELATED TO SEASON 8 Season Blood serum Variable Ca P Mg Zn Fe Cu Se Liver Mn rainy dry .290 -.209 .246 .169 -.080 .646 -.071 -.188 -.039 -.913**.002 .433 .109 .185 Zn rainy dry .302 .186 .409 .645 -.127 -.723* -.642* .554 -.268 .497 .405 -.024 .120 .540 Fe rainy dry .356 .530 .296 .787* .154 -.251 -.586* .907** -.143 .000 .251 .789* .240 -.042 Cu rainy dry .587 -.274 .063 .032 .425 -.803* .259 -.059 -.476 .730* .869**-. 536 .715* .552 Co rainy dry -.705*.078 .353 .036 -.485 .805* -.244 -.131 -.050 -.817* .755* .551 .353 -.091 Mo rainy dry .196 -.869*.115 .459 .469 .156 -.010 -.537 .564 -.412 .049 .361 .127 .267 Se rainy dry -.238 .671 -.239 -.076 -.237 .062 .842* Bone Ash rainy dry -.518 -.384 .864** .564 .346 .468 .468 -.841* -.155 -.354 .314 .534 .024 .125 Ca rainy dry -.463 -.397 .673* .491 .428 .377 .329 -.785* .204 -.353 .170 .525 .481 .255 P rainy dry .155 -.220 226 147 .850**-. 271 -.130 -.610 -.211 -.211 .271 556 .310 .365 Mg rainy dry .487 .317 845** 441 -.491 -.322 -.445 -.585 .169 .021 351 106 .062 .675* Correlation coefficients seasons in liver and 8, based on 8, 6 means 7 means for rainy anc for rainy and dry in bone. dry *P<0.10. *P<0.05. **P<0.01.

PAGE 144

133 TABLE 30. SUMMARY OF ANALYSES OF VARIANCE OF SOIL MICRO-MINERALS — MEAN SQUARES BY SEASON Response Farm 3 Paddock/ farm Residual Rainv season Boron .0001 .0104 .0094 Copper 11.0098 6.1491 3 .2721 Iron 362715.97* 113986.02 39531.26 Manganese 296.88 138.29 10.29 Zinc 6.8931* 1.5172 .2933 df 2 12 Drv season 44 Boron .1016** .0118 .0044 Copper 5.9755* 1.7999 .2017 Iron 837900.09* 178337.50 41748.07 Manganese 70.8478 50.7362 4.8890 Zinc 2.9718 1.9128 .3450 df 2 13 38 "Test farm using paddock/farm as an error term. Represents degrees of freedom. *P<0.10. *P<0.05. **P<0.01.

PAGE 145

TABLE 31. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE MICROMINERALS — MEAN SQUARES BY SEASON 134 Response Cobalt Copper Iron Manganese Molybdenum Selenium Zinc df b Farm 3 Paddock/ farm Rainy season .0440** .0951 10611.83 10420.08 4 304 2 4002* 0186** 12 .0047 21 .6271 17280 .00 29354. .30 1. .1388 •/ ,0029 150. ,87 12 Dry season Cobalt .0021 Copper 3 .8044 Iron 2303 .26 Manganese 233382 .23** Molybdenum .3952 Selenium .0017 Zinc 231 .4363' df 2 .0045 6.3543 5601.09 33671.88 .8585 .0096 71.2156 13 Test farm using paddock/farm as an error term. Represents degrees of freedom. *P<0.10. *P<0.05. **P<0.01. Residual .0021 5.2256 1972.74 3709.55 .2380 .0031 52 .4691 52 .0091 2.5376 3521.46 7399.76 .1740 .0071 63.5348 49

PAGE 146

135 TABLE 32. SUMMARY OF ANALYSES OF VARIANCE OF BLOOD SERUM AND LIVER MICROMINERALS — MEAN SQUARES BY SEASON Response Serum Farm" Class" Farm x class Rainy season Residual Copper Iron Selenium Zinc df b Copper Iron Selenium Zinc df Liver .7717 5.5228 .0738* .2370* 2 .6604* 9.8931* .0155 .1519 2 .0063 1.4429 .0086 .1566 2 .1762 2.1731 .0079 .0301 3 Dry season .4143 .8614 1 .0035 .8949 .1284 .4742 .0049 .2649 Rainv season .0384 .4120 .0033 .0352 85 .0709 .5642 .0011 .1394 105 Cobalt Copper Iron Manganese Molybdenum Zinc df .0333 12013.36* 166259.84 94.7558* .0426 2077.01 .0337 1829.60 380856.81 57.6226 .0763 5703.33 .0240 2220.49 150557.0 17.0931 .2365 3208.08 3 .0216 2169 ,16 5, .9142 ,1411 410. ,18 42 Dry season Cobalt Copper Iron Manganese Molybdenum Selenium Zinc df .0676 32103.09 152996.83 120.3429 .0018 .0253 4489.88 2 .0119 3195.19 618011.81* 10.8961 .0014 .0646 3386.23 2 .0007 903.17 4479.49 14.8920 .0004 .0862 642.80 .0031 4147 .45 53849 .97 11, .6766 .0003 ,1452 899. 72 61 Test farm and class using farm * class as an error term. Represents degrees of freedom. P<0.10. *P<0.05. **P<0.01.

PAGE 147

TABLE 33, 136 SUMMARY OF ANALYSIS OF VARIANCE OF FORAGE (PARTS) BY SEASON TEIN ' IV ° MD AN ° "^O^NEKALS-MEAN SQUARES Response Part" df° Calcium Magnesium Phosphorus Potassium Sodium Crude protein IVOMD df Paddock * part/ farm Calcium Magnesium .5632** .0332* Phosphorus Potassium Sodium .0021 .5131 .0000 Crude protein IVOMD 828 .4354** .0000 1.0320** .0530** .0099* 3.5430** .0023 845.0289** 1272.5860** Rainy season .0212 .0041 .0044 .2868 . 0O02 25.1014 109.7614 12 Dry season .0542 .0008 .0011 .2538 .0018 12.5668 48.7012 12 Residual .0700 .0079 .0578 .4859 .0005 16.6603 55.8628 85 .1563 .0013 .0016 .2230 .0003 6.5930 22.0784 82 Test plant part (stems, leaves) using paddock * part/farm. 'Represents degrees of freedom. P<0.10. *P<0.05. **P<0.01.

PAGE 148

137 TABLE 34. SUMMARY OF ANALYSES OF VARIANCE OF FORAGE (PARTS) MICROMINERALS— MEAN SQUARES BY SEASON Response Cobalt Copper Iron Manganese Molybdenum Selenium Zinc df b Cobalt Copper Iron Manganese Molybdenum Selenium Zinc df Part" .0010 9.0807 10617.96* 6780.05 14.8481** .0059 306.7934* 1 .0004 1.8003 18280.45* 128730.53** .0073 .0080* 202.0959 1 Paddock * part/ farm Rainy season .0011 17 .7774 3121 .55 5748 .89 .3028 .0055 66 .5135 12 Drv season 0046 1 1553 2712 29 9106 28 5432 0018 74. 5792 Residual .0048 7.8907 2535.73 2742.53 .1814 .0034 51.8941 85 .0058 2 .7159 5060 .09 8508 90 6608 0023 50 1032 12 82

PAGE 149

o Eh Q W Eh 5 < Eh ss a o 2 o u c > 3 Eh o~ K CO Bi H w < Q CO a a x u a a < w a « m H z u h w 2 p. O CO « U Q § < C O < CO « <: o w ft, en 133 mioioiOiOimi tl cn in o (N OO rMtN OO OO OO com Hi OO OO oo o'o 0*0 Oh in KS §5 22 3= £2 „„ „, OO mm OO OO OO h o' cr ^ (N tN iiO mm OO *T O » ffl OO OO otN iam oo oh h(n mm mn OO H el OO OO OO 10 (N t-> V OO OO OO OO oo o ft om hot tot men ^ *j co rrsiN o io ho oo nh (nh o\r* OO oh oo oo oo om m^j nh rm i0^ OO mv Tm Om oh <** h ra. m rOO (Nh OO OO OO* r-H oh cm om f\im orcoror(N iD CO O Ol PH 3S Sg gg gg gg S« sg OO OO OO OO OO HO hh miN r-O ho OO INH 01 £T> t~JO OO ^ >-t OO OO OO *]
PAGE 150

139 S i V. *~ c u I "* m a 1 V f** 0) Q L> a

PAGE 151

140 re w 1 a H Ej O w O o u X o a o H H ~ U 03 <; h o < In a) « O 1 C 1 O i 6 i O 1 O 1 * u iO (N a s 3 ^ a\ co o — * s w o o n S M M o n o o e "*3 O O a a 10 H O i-l 3 S c »4 o o -i a 3 rib T o g\ o O O o o o> o o co u £ £ o o a a o> m »N o a a> o u o o o o 3R K o o o o — T 0" O ffl ul oo oo ~ a co a> o o o o I o o o o o * oo Mt>n a> H^i 5 3 woo oo u o oo oo" do" O O in^f o a! --00 cn\o Of* £ 10 OO cn*r Cfi^D n vo MO fO •* f* o o --H d o o m in sjrf!''3jJ3!rf6 , '3&iB HTJ Ij-O H-O M-O H "O W-3 1--0 I I I I I I sis 8 i i i s i

PAGE 152

141 o — K >. — TJ Sn SI i-» ^ CT> Ji OO —• O I I i-l ffi BlTi D IO (T f* ff ^ . fl c u i nj a arj) in ^ ui mo* n co — tr> fl — O OO On OO O in -hO O rCO ro O CO 3 o — . i _ -o b e >0 rN O <0 O ** •o >> 2 a\ o oi m o o cm m co -i C 1u o 01 oi oi ai oi ii mi oi a\ cm a * 4 c s. .. 41 M»1 i-L t* HI M I . IB fr iw -i O r»o O O u a o 1c CM -O OO CO ^ l " JIN O(-1 M U 1J 1« T3 M OO OO OO OO 1 1 OO OO a . — § fl V c a I IS M B oj CT< 0^ CM n O >-i in CT* H O OrOO COCO CO \0 — O OO ;r>^D 3 U a w rH « •ho oo o in cr>in oo oo ^ w *0 u c i! 01 u 4 s i-i IN \0 ft r\C V O c • E 0. £ (Din oo r-n orn oo OO o cm • Q e 41 W B >.>>.>*>.>.>. •8 ^ c c s s a c a •u H >, -H >. -H >, -H >. -H St 'H S — >. flu ---. it ki .. a U n u «u V c s» S k"0 ~ — ~ r -c ki-a c w D>-< g 3 — 8 u u o -* o-3 i.-. v U (J (J "O o •n o ij co 1 u > ro O o >0 o o 4) -• 1 j: .. . iu — ° u C < IX) c ta id h -h io ki O X) C) 4J W 41 — 4) 3 V W 3 ~h in fl *i tr *D co -h 5 H U Ol H < i I s 1 a g a a a a, a a a E S 5 S S & S Means H. Ian for D. Standa NRC (1 McDowe 1j 1 3 1> C Q 41 E O £ £ £ n H 10 .Q TJ

PAGE 153

142 O ** O O O ffi w w H u w En H o H c u g o o o o o o c o rc h o" o o o o a .O" o o o a o o o o ro § § O 10 O»0*ra>vDp4OO c s IJ • Hi — > < O CQ rr o O O CTi in n» o o ^ o ^ o o o rv tfl — C t-t O O O O O o to •* co a en v O O O -^H H o o = Si a a & a a S, a q t e a • c * >

PAGE 154

T, O H En < ci o u 143 w 2 H s: o « u H as p •z < a §ra w • S z H O 5" CO O < « W u co «! s o H a a aw Eh -< a. j w w E-i S E-i < CO S eC u co H E-I < m a h K U O H b hj t. H U o o CO u i O f» m r-i O O ric •* aj a co I 10 o (N ^ o* ^ r» into a} t it ; oo r-f-cno o « Of IV© f*> r\Q ffi (Ti a h -. a, n g I 12 SI a o 3£ S3 S

PAGE 155

144 1 O o o £ H H a o M En O CD i s o o c w o o 5 § — . " O tO H CO (N 3 o ^ o (N m c o cc C « m 2 H O CO CN CO O O CO -. H o O AH H O in r— H u J, ,' ," ,' "• O S C+ orto ico o ^ ^ j w c a to wo <*> O W O (N PI O O r\© \o o A n *4 in Q H rt * r * J O « U O O ^ co -h rv O <*• O ON IN iC V Q o m f« oi oi >n o W — pn o -> o. Z c 0) M (0 c c * • 3 J s J 2 < O w rt £ 41 >= 2 O r10 PI i-> fN *T fl 11 II >. ori0 w r-j « o oo rr— i m eo O *N PI PI 10 PI Pi 41 z w Q >• b H CO * « + •o e; O r-iM cno — i Oi in o r~ — i o n i-ioi \s o o 3 O in ifl V CO INrt CO o 10 m «»tM (no ?n o cow r-co co— i pi o oo w^h pio r> = « Eh u -ii '' >c H Q I £ W Eh 0*-i o co in in m oi — i O— OCD IT, PI MO --< £$ I if m r3 ifl W ON in O CO -J X 1/1 4] O01 O Om mo 10 o -«co rrVia to™ 3 o o tn "" mo -h w pO C0(N tDO enw OO ~ O iNv o« too pn pi m o Hin ow pio riw coo eo O n h cow !Dr(no ai o "in ry ow r^iji wco in — i v in r*m ro% -_ u i >0 — i pn oj o w rw Of" O Oio vow wpi (N — i ow in n p> m c W m Eh f " E § CV _ m 3 3 ui eg EO— i-M 4) 5J3C 3 .E to tfl E H e — .._ ; x 3 $ ib — fi u „ -: .r i : ? t « >. « *j Si! 1 i] h .„ S Sic — —:..' -. nj — j^C4) 'j icm — ^ c oi uia >» j-i 10 • . r* — \ O m o 41 -h o h • O 5 -"O cr*j -oa o e c ja -« « o «jj3 ai o 00 i-m-4 Ocu O o. so, mo w X nu £ co — 4 9 i *e -tsa oc cjj --, o fl£ ieo oo fc-n •-o 04» oa. i p. wo m£ mo £w a. o ? v a, o. o. *

PAGE 156

'-•• APPENDIX C RAW DATA

PAGE 157

SOIL DATA CODES OBS = Observation number SEASON 1 = Rainy (1987) 2 = Dry (1988) FARM 1 = San Jorge 2 = Don Benito 3 = San Francisco PADDOCK 1 = Nevado 2 = Papayo 3 = Regadera 4 = Esperanza 5 = Tablon Alto 6 = Pantano 7 = Laurel 8 = Mirador 9 = Siberia 10 = Pantano Amarillo 11 = El laurel 12 = Aguasal 13 = Porvenir 14 = Panama 15 = Llanos 16 = La Sierra 17 = Palocaido 18 = La Mina 19 = Cajon 20 = Puertas 21 = Buenavista 22 = Lomaciro 23 = El Pantano PH OM AL CA P MG K Soil pH Soil organic matter, % dry matter Soil aluminum, ppm dry matter Soil calcium, ppm dry matter Soil phosphorus, ppm dry matter Soil magnesium, ppm dry matter Soil potassium, ppm dry matter 146

PAGE 158

147 NA = Soil sodium, ppm dry matter FE = Soil iron, ppm dry matter cu = Soil copper, ppm dry matter MN = soil manganese, ppm dry matter ZN = Soil zinc, ppm dry matter B = Soil boron, ppm dry matter

PAGE 159

dbs seas rm mi ?m 3 10 13 :4 :s 17 is 19 23 24 25 26 27 2B 29 30 31 32 34 35 36 37 39 3' : 40 41 42 43 44 45 46 47 48 49 1 3 I 3 1 3 I 3 1 3 13 13 13 16 16 16 !6 17 17 IS 19 315 I 5.0 16.0 I 4.9 13.1) 365 4.8 26.) 387 ! 5.4 5.0 27 5.1 5.:) 31 5.2 4.0 54 5.3 4.0 S3 5.3 5.0 63 5.4 6.0 36 5.6 6.0 5.3 4.0 63 5.3 4.0 54 5.2 5.0 «] 5.0 17.0 270 5.0 15.9 193 5 1 19 '3~ 5.1 !5.0 152 4.7 J9.0 8:3 4.5 23.0 1053 4 .5 16,8 963 4.5 22.0 1053 4.5 20.0 765 • c 25.0 373 4.7 25.0 634 4.5 15,0 535 4.6 15.0 553 4.5 15.0 575 4.5 19.0 535 4.5 14.8 566 5.3 13.0 108 5.2 18.0 162 5.4 20.0 39 5.4 21.0 M 5.! 29.0 315 5.1 30.0 252 5.1 23.0 270 5.2 32.0 297 5.2 23.0 297 5.0 32.0 360 5.1 30.0 306 5.1 32.0 225 J .3 30.0 333 5.1 32.0 315 4.8 32.9 436 4.9 30.0 360 4.3 32.0 360 5.0 30.0 270 3*4 352 m 1308 554 1103 1.26 i5u2 1524 1050 1184 1164 1154 424 510 466 454 104 112 20.4 30.0 18.3 236.8 229.2 432.0 257.2 35.1 50, 44 46 18 53 86 116 100 400 212 »26 516 730 10 00 602 620 514 380 274 262 200 482 314 29 9 11 9 5 :oo 90 150 100 21 15 10 17 68 110 110 30 55 34 10 242.4 "13.2 253 7 273.6 304,8 334.8 357.6 31.2 46.8 23.8 26,4 18.0 19.J 15,6 13.2 13.2 15.6 13.2 12.0 12,0 14.4 20.4 10.8 43.2 56.0 34.8 76.8 44.4 39.6 26.4 28.8 31.2 22.3 25.2 3.6 10. 8 13.2 19.2 13.2 8.4 9.6 105.3 137.2 123.7 148.2 52.4 53.5 74.1 50.7 .01.4 23.4 39.0 7.3 62.4 101.4 78.0 37.5 56.3 133,3 31.3 78.0 35.8 124.3 105.3 37.5 74.1 128.7 117,0 152.1 206.7 81.3 81,9 89.7 62.4 62.4 91.9 62.4 27,3 46.3 39.0 62,4 46.8 42.9 46. 8 148 W FH 3 «N ZN 9 13.8 325.5 7.2 6.2 :.7 0.22 25.3 461.4 0.8 5.5 2.1 0.22 23.0 230.0 C.6 5.4 1.4 0.22 13.9 373.5 0.5 2.5 1.0 0.13 43.7 150.5 0.3 16.3 2.2 0.32 46.0 395.0 1.7 23.5 3.3 j.27 43.7 4O6.0 2,4 32.7 4.2 0.27 50.6 573.0 2.6 33.0 2.3 0.27 27.6 725.0 2.1 11.5 3.5 0.36 34.5 559.0 2.4 14.7 3.7 0.22 36.3 586.0 3.0 17.3 4.3 0.41 52.3 585.0 3,0 14.7 3.2 0.22 25.3 583.0 3.4 21,3 3.7 0.27 59.8 515.0 3.1 12.6 3.1 0.41 48.3 713.0 1.9 10.6 3.0 0.35 48.3 456.0 1.6 3.2 2.5 0.22 16.1 273.0 1.0 3.6 1.6 0.32 13.8 231.0 0.7 4.1 2,7 0,41 11.5 170.0 0.6 3.3 1.7 0.32 9.2 338.0 1.2 3.0 1.6 0.22 2.3 35.0 11.2 1.4 3.0 0.182.3 870.0 11.8 2.5 2.1 0.22 2.3 670.0 1.0 3.3 2.4 0.32 2.3 1150.0 1.1 4.3 2.0 0.22 2.3 1160.0 2.2 2.9 2.9 0.18 2.3 1200.0 2.0 2.2 3.7 0.41 2.3 450.0 1.7 2.3 2.3 0,09 2.3 140.0 1.3 1.7 1.9 0.36 2.3 350,0 3.3 7.3 0.27 2.3 640.0 1.5 7.2 2.9 0.36 2.3 750.0 2.8 3.0 4.7 0.27 2.3 300.0 1.9 4.5 2.6 0.27 13.8 3*8.0 1.2 11.4 3.1 0.32 16.1 401.0 2.2 10,0 3.3 0.22 6.3 368.0 1.9 6.9 2.7 8.27 2.3 136.0 1.2 12.9 3.6 0.32 2.3 331.0 1.0 7.3 2.5 0.32 4.8 863.0 l.S 12.5 2.5 0.46 6.9 749,0 0.7 4,6 2.0 0.27 2.3 593.0 1.8 4.6 1.9 0.32 2.3 679.0 1.3 7.2 1.8 0.46 2.3 766.0 1.2 7.6 2.1 0.27 2.3 572.0 1.3 9.2 2.5 0.22 2.3 251.0 0.4 3.4 1.5 0.27 2.3 310.0 1.0 5.4 1.8 0.32 2.3 231.0 0.7 2.5 1.5 0.04 2.3 449.0 2.3 3.3 3.1 0.27 2.3 369.0 .4 5.4 .7 0.27 2.3 256.0 ».6 2.9 .8 0.13 2.3 381.0 .5 1.6 .4 0.32

PAGE 160

03S 5EAS FASH ?A3D ^H OH 51 52 53 54 cr 56 57 53 59 50 61 52 53 64 65 66 67 68 69 70 71 72 73 74 75 76 77 79 79 90 91 82 63 34 85 36 87 38 89 90 91 32 93 94 95 36 97 58 99 100 2 1 19 19 13 :9 20 20 13 13 13 13 14 14 14 10 10 10 15 15 15 13 19 19 19 17 17 4.8 30.0 5.0 26.0 4.9 25.0 4.9 23.0 5.1 5.1 1 5.0 15.2 1 5.0 6 5.1 6 5.5 7 5.1 8 5.3 4 5.7 5.7 5.0 5.5 5.3 5.3 5.6 5.3 5.5 5.3 5.9 5.8 5.3 4.8 4.7 4.8 4.4 4.3 4.5 4.4 4.3 5.0 5 2 4.9 4.8 5.0 5.1 4.9 4.8 5.2 5.0 4.8 4.8 :3.o 19.0 2i. e 24.0 25.0 25.0 24.0 25.0 23.9 23.9 23.9 12.9 7.6 7.5 5.6 5.6 7.3 7.0 4.5 3.1 4.3 4.3 3.6 4.8 22.7 24.6 21.4 22.0 16.7 19.2 19.7 20.8 22.0 15.8 18.7 16.3 17.2 17.7 23.2 21.4 19.7 19.7 29.1 28.3 27.5 486 270 306 315 206 261 243 261 90 :ob 108 12S 315 329 477 1080 810 738 810 360 378 306 144 162 108 360 315 450 360 351 531 531 62 152 40 26 52 378 490 556 524 258 298 464 25B 194 170 254 640 1514 1490 1320 1814 1616 141 1740 1119 1400 926 1119 1532 376 730 624 70 74 72 40 76 14 110 54 156 124 810 230 232 9 15 5 5 5 44 5! 66 57 14 7 14 45 24 :e 25 25 30 19 13 10 19 15 17 38 12.0 14.4 8.4 7.2 26.4 30.0 24.0 23.8 20.4 13.2 15.6 16.8 14.4 19.2 8.4 25.2 243.6 414.0 194.4 333.6 186.0 1B0.0 180.0 363.6 132.0 253.2 212.4 2*9.6 25.2 14.4 19.0 16.8 18.0 15.8 8.4 6.0 6.0 6.0 7.2 6.0 6.0 20.4 12.0 16.8 20.4 28.8 14.4 20.4 58.5 50.7 35.1 31.2 39.0 105.3 140.4 37.5 113.1 124.9 46.9 52.4 58.5 31.2 54.6 62.4 39.0 167.7 128.7 132.6 128.7 202.8 187.2 163.8 132.6 101.4 97.5 124.8 113.1 58.5 46.8 "' 7 21.2 58.5 42.9 46.8 13.5 15.6 11.7 11.7 19.5 23.4 31.2 50.7 27.3 58.5 58.5 42.3 81.9 70.2 149 NA FE CO m n b 2.3 398.0 0.8 4.5 2.0 0.19 2.3 2';4,3 0.3 4.2 1.6 0.32 2.3 261.0 0.3 2.1 1.2 .1.46 2.3 348.0 0.5 2.2 i.l 0.22 2.3 315.0 0.4 2.3 1.1 0.46 2.3 331.0 j « 5.4 1.4 0.18 2.3 362.0 1.2 3.5 1.6 0,5' 2.3 439.0 2.1 5.9 1.4 0.13 2.3 491.0 !.5 3.2 1.3 0.22 9.2 365.0 0.7 5.2 1.3 3.27 2.3 265.0 0.5 2.5 1.0 0.25 5.2 288.0 0.4 3.8 0.7 0.41 11.5 395.0 0.7 4.6 1.1 0.41 4.6 600.0 1.7 3.0 0.3 0.36 2.3 199.0 0.8 3.4 1.0 0.41 2.3 130.0 0.6 2.5 0.6 0.36 2.3 121.0 1.3 4.2 1.2 0.41 16.1 403.0 3.5 17.8 4.6 0.41 20.7 269.0 2.0 3,4 1.7 0.36 39.1 490.0 2.9 12.2 2.2 0.36 25.3 490.0 3.4 12.0 3.0 0.46 29.9 450.0 2.6 15.2 3.1 0.36 18.4 480.0 2.2 13.7 3.2 0.41 13.8 500.0 2.2 9,6 2.3 0.41 54.4 386.0 4.3 21.4 3.1 0.32 23.0 240.0 1.3 17.7 1.9 0.09 18.4 550.0 2.7 15.2 3.5 0.32 13.8 560.0 2.7 17.2 3.3 0.22 13.4 580.0 2.6 IB. 4 3.9 0.27 2.3 360.0 0.2 6.5 0.2 0.36 2.3 352.0 0.1 3.6 0.7 0.27 4.6 405.0 0.1 3.3 0.7 0.41 2.3 575.0 0.1 6.3 2.3 0.32 2.3 980.0 0.2 3.8 1.3 0.41 2.3 1050.0 0.1 3.3 1.2 0.41 2.3 1730.0 0.8 9.4 3.1 0.27 2.3 1500.0 1.1 3.6 2.8 0.22 2.3 320.0 0.7 3.3 2.4 0.27 2.3 430.0 0.2 3.5 1.6 0.32 2.3 260.0 0.4 4.2 2.2 0.27 2.3 210.0 0.5 3.1 2.2 0.22 2.3 170.0 0.2 3.2 2.0 0.13 2.3 320.0 0.4 3.5 2,3 0.27 2.3 90.0 1.6 3.8 1.3 0.22 2.3 150.0 1.6 3.4 1.0 0.09 2.3 170.0 1.8 4.5 1.1 0.18 2.3 175.0 1.6 4.9 1.1 0.13 2.3 260.0 1.6 5.6 1.2 0.27 2.3 240.0 1.2 3.2 1.1 0.22 2.3 270.0 1.0 4.9 1.2 0.27

PAGE 161

DBS SEAS FARM "ADD :0l 1 i 17 5,0 24.6 315 504 44 1J.6 !02 2 7 21 5.0 23.2 147 325 36 14.4 103 • 3 21 5.2 2S1 680 50 22.8 104 :u5 " 11 5.0 5.0 13.2 270 297 464 456 54 54 25.2 13.2 106 5 3 22 4.7 23.2 702 44 3 15.6 107 22 4.3 23.3 558 32 15 13.2 108 103 ) i 22 4.7 4.3 24.6 23.3 621 35! 214 74 13 < i 15.6 no 2 3 23 4.3 23.1 435 330 20 22.8 111 2 3 M 4.9 17.7 435 330 33 14.4 112 2 3 23 4.3 17.2 450 250 45 13.2 113 2 3 23 4.7 13.3 633 162 33 13. 2 150 K M FE CU IN IS 8 13.5 2.3 130.0 1.1 3.5 1.0 0.22 105.3 2.3 160.0 1.0 3.1 0.3 0.13 31.3 2.3 250.0 1.0 6.2 1.0 0.22 70 2 • i j . . . _ 4,0 0.27 31.3 6.9 150.0 0.6 4.6 1.0 0.03 25.1 6.3 253.0 0.3 :,6 1.0 0.16 42.9 2,3 25 q 0,3 5.6 1.5 0.03 1 ~ :.: .35.0 0.3 4.4 1. ' 0.27 23.4 2.3 125.0 0.6 4.3 1.1 0.27 J J. 1 2.3 47o,o 0.7 3,3 1.2 0.13 23.4 2.3 370.0 0.5 5,3 1.5 0.22 19.5 2.3 250.0 0.2 4.3 13 0.27 31 .J 2.3 170.0 0.5 3.0 0.36

PAGE 162

151 FORAGE DATA CODES OBS SEASON FARM PADDOCK SPECIES PART = Observation number 1 = Rainy (1987) 2 = Dry (1988) 1 = San Jorge 2 = Don Benito 3 = San Francisco 1 = Nevado 2 = Papayo 3 = Regadera 4 = Esperanza 5 = Tablon Alto 6 = Pantano 7 = Laurel 8 = Mirador 9 = Siberia 10 = Pantano Amarillo 11 = El laurel 12 = Agua Sal 13 = Porvenir 14 = Panama 15 = Llanos 16 = La Sierra 17 = Palocaido 18 = La Mina 19 = Cajon 20 = Puertas 21 = Buenavista 22 = Lomaciro 23 = Pantano 1 = Anthoxanthum odoratum 2 = Holcus lanatus L (native) 3 = Pennisetum clandestinum 4 = Tri folium repens 5 = Festuca arundinacea 6 = Dactylis glomerata 7 = Holcus lanatus basyn (imported) 1 = Stem 2 = Leaf 3 Whole plant

PAGE 163

152 CA MG K NA CU IVOMD = Forage calcium, % dry matter P = Forage phosphorus, % dry matter = Forage magnesium, % dry matter = Forage potassium, % dry matter = Forage sodium, % dry matter = Forage copper, ppm dry matter FE = Forage iron, ppm dry matter MN = Forage manganese, ppm dry matter ZN = Forage zinc, ppm dry matter co = Forage cobalt, ppm dry matter M0 = Forage molybdenum, ppm dry matter SE = Forage selenium, ppm dry matter cp = Forage crude protein, % dry matter = Forage in vitro organic matter digestibility, % dry matter

PAGE 164

OSS SEAS FAR PAD SPE PAR CA *G % '*N ZN DC 153 l• IvOHD :o i i 1 i '. ".' 1 i t i ! : i 1 * 1 i 14 1 i il 15 J i ;5 1 1 £ 17 i i L sa 4 1 1 _ 26 2S 4. J 30 3; 32 ! H f j j * j4 i 3' 38 39 40 44 45 46 47 48 1 i 1 1 1 1 i 54 1 36 57 3 4 3 3 3 3 3 5 3 5 3 5 3 3 3 3 4 6 4 S 4 6 0.13 0.30 0.26 0.13 0.23 n ?•? 0.27 0.35 0.16 0.14 0.28 1 0.17 -i ft *5»l L i'iJi 3 0.37 1 2 1.90 3 1.81 1 0.18 0.22 0.20 0.42 0,66 0,64 0.70 1.3S 1.96 t jn L.ti. 0.32 0.39 0.21 0.57 0.65 0.44 0.41 n *>c •J 1L 1 0.13 0.21 0.18 0.29 0.30 0,12 ;"! 3? L. Li ml 24.3 23.9 19.3 42.1 23 i 23.6 50.6 42.7 57.7 32.3 70,1 38.5 28.6 28. 1 43, 41.7 46.2 32.8 40.4 23.7 25.0 26.4 32.4 31.0 30.8 30.6 ';Q c 0.09 0.10 0.10 0.06 0.05 0.03 0.07 0.07 0.05 0.07 0.08 0.06 0.06 0.07 0.36 0.18 0,21 0.16 0.18 0.13 0.12 0.11 0.10 0.14 0.24 0.47 0.26 0.23 0.11 0.08 A 1 C it . I .1 03 0.16 0.11 0.13 0.09 0.08 0.03 0.11 0.07 0.03 0.16 0.03 1.19 1.70 1 ' ~: 1, .'8 2.25 1.38 l.i./ 1.38 1.74 2.27 1.37 , n 0.60 0.31 0.41 0.11 0.05 0.03 0.24 0.16 0.15 0.33 0.71 0.45 0.62 0.17 0.03 0.22 0.56 0.50 0.75 A 50 0.13 0.88 0.16 1.07 0.40 0.34 0.21 0.66 0.63 0.84 0,96 0.38 0.33 0,04 0,07 0.04 0.07 0.05 0.04 0.06 -. I c *.'« id 0.06 0.07 0.05 0,18 0.15 0.10 0.12 0.15 0.18 0.13 0.09 0.20 0.33 0.13 0,15 12 0.17 0.14 0.05 0.09 0.10 0.10 0.03 0.02 0.11 0.09 0.18 0.04 0.07 0.04 0.07 0.07 0.07 0.07 0.05 0.21 0.10 • 2 25 •--c 02 07 04 £.5 41.8 20.3 61.3 15.3 57.1 7,9 44,2 25.5 55.7 v: a ~a_" A ". 'A 7 i II.! 11.5 15.6 11.3 15.3 16.4 15.1 16.7 12.4 15.9 15.4 14.2 14.3 19.2 34.4 29,0 20.0 21.0 15.7 23,6 18.5 13.2 19.4 18.8 21.4 30.7 27.2 19.9 20.5 I 1 + .lii 17.4 16.8 15.8 16.0 18.7 1 c Q 22.9 69.7 58.2 59.7 55.4 62.5 64.3 65.4 66.1 63.2 49.0 73.6 62.2 75.3 40.3 40.4 67.6 57.3 69.5 44.0 47.8 53.4 57.9

PAGE 165

154 DBS SEAS r^. PAD 3PE PAR 24 P 13 ^ M cu FE J N :n 13 : J 3C CP IV0HD 52 i 1 4 3 i i 0,13 0.47 0.15 4.24 0.05 16.07 244.6 59.9 51.3 A --:C 59 1 1 4 3 2 0.30 0.22 0.21 2.48 0.03 12.20 SI. 9 99.0 40.2 0=46 0,12 10.9 41.6 -) i i 4 3 0.34 0.29 0.24 2. 73 0,04 10.01 66.3 119,8 46.6 0.2S 0.16 '• * 1 10.8 -2. 1 61 I i 4 7 4 0.16 ;"! 70 0.12 2.08 0.03 3.11 26.8 166,3 28,4 ;}.06 A .-': 0,01 -. A 7 it, 4 :4. 4 i 4 2 y.yj 0.42 0.17 3.3b 0. 03 6.8B r*7 i J.' ; t '• "-''-. _ H 1 b . A 0. -.'-* ft "T.4 -J . / t 0,13 13.7 70.1 53 1 i 1 : 0.40 0.40 0.13 '": (] ' 0.03 3.01 73 ! * -.1 c; 23,3 . •) / 0.23 0.09 17.6 ji 1 4 1 . , , , , , , . , 0.05 -.; ;1 1 J 4 i . , . , . . , , a , 0.0B J^o I 56 4 1 4 4 7 1,59 0,29 0.38 4.25 0.13 3.21 33,3 33,9 34.1 0.08 , 0.09 23,3 67 1 i 4 4 i . , . , . , , , , , , 0, 12 19,2 bS 4 1 4 ~ i 0.33 0.31 0. 22 7 C7 0.02 8.49 68,0 159.7 23.4 1 ^ 0,23 0.17 13.7 51.5 69 1 1 4 3 0,35 0.23 A 7S ftc. 0.03 8.72 76.6 135.5 27.1 ft 1 7 0.25 0. 17 14.0 47.4 71 I 1 4 .2 L 0.40 0.12 '• 7 c 3.13 9-02 8.01 75.8 71,5 30,4 0.18 0.03 0.08 13.8 i 7 ti. ; 72 1 i 4 U 0,05 0.24 0,06 0.70 0.00 1.74 11.9 10.9 24,5 0,07 , 0.17 4 4 4 45 6 73 1 1 1 -^ 3 J . . . , . , , ( , , , 0.08 18.0 74 1 1 4 7 2 0.53 0.18 A. 7 A . 40 2.31 A A7 3.13 32.2 159.7 23.7 0.10 0.36 0.07 14.1 50.9 " i ( 4 3 3 0.21 0.32 7 7 1 3 0,03 5.27 258.7 264.4 01 1 *.i 1 i 0.06 1.15 , , "-_ 1 i i ~ [ 4 0.30 0, IS 0.09 0.34 0.03 1.19 255.6 238.6 13.3 0.03 1.30 0,13 9.2 45,4 SO 1 i E J I 2 0.34 7 k u« 12 2.72 0.04 7.24 153.1 245.9 28.0 0.03 1.44 0.02 L t a J 55,5 31 1 i 5 i 2 0.40 0.24 5.14 2.66 0.03 7.04 135.7 215,7 25.7 0.03 1.78 0.06 18.5 64.1 02 1 '. r 3 i i 0,45 0,29 0,17 3.07 0.05 7.29 200.3 230.2 26.3 0.02 2.08 0. 04 21.7 67 8 33 i i < c 1 0.43 0,26 'J. lb 7 o 1 ; La Ji. 0.04 4.45 291.5 7r I 23 1 0.02 2.10 0.06 17.1 S3 7 54 4 i E J 1 2 0.42 0,27 0.21 3.50 0.06 4 A i 1 tdVi L '1^ ' 31.0 0.01 1,63 0.14 23.6 71.4 [ C J 1 7 0.38 0.23 0.19 2.54 0.05 7,26 239.3 23.8 0.02 1.94 0.10 23.4 57.9 101 i X 2 3 7 1 O.il 0.24 0.07 0.04 5,87 125.7 275,1 25.8 0.03 0.18 0.25 , 102 4 i ~ 7 2 0.20 0.28 0.03 2.32 0.02 5.21 127,6 153.6 16.9 0.01 1.75 0.20 24.1 7*3 c i V J \ I 2 3 7 / ^ 0.25 WiUw ft i 7 3.34 0,05 7,43 345.5 77 i r 28.3 0.01 2.02 0.15 21.3 105 1 7 3 7 2 0.24 0.40 0.10 3.57 0,03 5.76 114,2 177.6 70 ft n,01 2.03 0.13 25.8 :og 1 9 -i 7 > :'\ 7 1 0.35 0,03 3.17 0.03 5.34 123.5 239.5 I 7 . 6 A Art 1.27 0,10 20.3 107 1 2 9 7 i 0.10 0.25 J i -.* " '.L 0.02 4.23 -'J. L 234.5 26,0 0.01 0.33 0.14 14.1 ioa 1 <7 :7 2 O c 0.34 0.13 3.65 0.05 7 h7 121.3 219.5 23.9 0.01 3.70 0.06 •7t; 7 75.7 103 J 9 0.26 0.34 0,11 3.46 0.03 6.36 155.0 252.7 20.8 0,01 1.89 0.18 20.0 T A J ! 1 1 4 2 ? " i g 1 1 , 37 0,07 3.39 0.03 4.60 76.7 303.3 19.8 0.02 "• crc; 0.09 , ' 1 1 1 2 9 7 -: 0.24 0.41 0.11 3.65 0.05 3.29 93.4 23 ' ' 19.7 0.02 3.62 0.06 ^ "i 4 71 ~ 112 J 2 9 7 rt 7<7 0.26 0.11 7 J^ 4. '4 0.03 5.04 71.4 257.2 17.3 ?;! 4.24 0.07 19.3 r C 7 113 i 2 10 •7 i i A A;7 0,26 0.10 2.85 0.02 5.47 -ic 7 258.6 7C « 0.02 0.34 0.07 i i . U 114 1 2 10 7 _ _ 22 0.30 0.17 3.28 0.03 6.49 94.1 231.4 28.3 0.01 2.25 0.14 25.7 115 i 2 10 " j 0.20 0.24 0.15 3.07 0.02 g f.7 108.7 77i j JUT. i 30.5 0.01 1.83 0.18 21.6 57,7 1 15 : 7 10 2 i 0.13 0.28 0.11 3.53 0.05 20.09 459.0 307.7 42.1 A A7 y , v j ft 77 0.13 t * 1 7 ill 1 i •7 10 7 '2 0.27 0,30 0.16 3.79 0.05 7,27 296.5 233.8 £ 9 . 1 0.01 1.54 0.22 24.0 63.4 118 1 i 10 3 0.24 0.31 0.14 3.43 0.04 11.63 143.9 360.4 «r 7 J J. 4 0,01 1.38 0.10 21.6 53.5 119 i 7 L 10 1 0.12 0.32 0.11 3.73 0,04 12.20 153.3 346.0 45.5 f\ ft -] 0.22 0.10 , 120 i •7 10 2 C.24 A ta 0.17 3.91 0.03 7.56 36.8 274.5 29.5 0.01 1.54 0.09 23.8 121 \ -7 L 10 _ 7 _ 0.22 0,30 0.16 3.46 0.02 6.69 143.0 315.3 29.2 0.01 1 "3-7 0.07 0-7 A 59.2 i .-^n i 2 10 _ *i 0.27 J. 4A 0.13 2.95 0.03 6.63 101.3 225.2 29.2 0.02 1.04 0.07 , 124 i •7 io 7 3 0.29 0.32 0.14 3.06 0.03 5.24 92.2 25.2 0.01 1.73 0.17 23.3 70.3 125 i 7 4 i 7 t o.oe 0.33 0.08 3.72 0.02 13.80 156.3 277.9 53.0 A A 7 0.14 0.11 15.7 125 i 2 11 7 _ 0.20 0.38 0.15 4.48 0.03 7.29 77.7 232.4 23.3 0.01 1.49 0.07 25.9 77.1 127 i 2 i * 7 n 0.21 0.42 a 4 r 4.70 0.05 7.47 57.5 231.6 25.9 0.02 1.32 0.11 24.4 7 7 -i

PAGE 166

155 fi o ; SEA -^ ?4D SPE ?A3 CA : : IS H NA L-U 77 *N ZK CO m :2 C? IVOHB [20 | z i 4 7 I 0,10 0.30 ", no ^27 0.03 3,98 i 50 1 3 259.8 22.5 0.02 0.18 0.09 121 1 1 11 7 4 0.13 0.44 o, ih ": ' 0.02 10.42 33.1 113.4 20,8 '' A t 1.3S "-. -3 22.7 7Q 5 30 4 J. 1 1 7 j 0.16 0,36 u. 12 3.44 0.04 11.15 95, S 2 1 8 & ib. 0.02 0,53 0.10 13.4 7* 2 i a a 7 jj 7 2 0.27 0,34 ft ! ^ V f i J 3.63 i AT 7,22 ^j,/ : "'5.3 22.4 0.02 0,35 ft *' Q 2!. 5 7c a * "T i i 11 :... . . -1 -j * n 3.27 0.01 5.3! 32.3 21.6 0,01 0.40 f~\ '.' '? 4 A, 7 i j ; 74 3 " ':! 1 2 4 j 1 3 . ! P 0. 23 0.03 3.83 0.02 10.27 1 4 3. 3 !L0 29.8 ft .-'. 4 0.15 0.08 IS, 2 i 7 < t 7 _ 0.34 0. 33 0.1S " : '•--. 'J » ''J2 5.30 78.3 * * 1 25,1 0.02 1.84 0.17 25.9 78 2 J 2 u 7 ft V) ^S n ' ~: 4, dU 0.02 7.24 33.2 253,2 2b. S 0.01 0.57 0.10 22 . 75 7 7 4 i "' i 4 ) . i 3 f ! J 7 \) '^ 2 1 iO 0.02 10.6, 227.7 202.6 21.4 0.02 0.31 0.C4 33 J 2 i 1. Z 0.31 •j , 2 4 0.16 3.17 0. 02 .7 " ' 35.3 133.5 20.0 a QQ * 7C 1 . / J V 1 v'3 20. 1 1 ^ * 4 2 i"/ 2 ? n 7. ;' * 7 ' :" j = *5 77 0,02 3.22 IIP f\ 178.1 7 7 0,01 1.18 0,20 16.4 ;"•, HO I . * -[ 2 4 ' j 1 4 ^ 0.36 ^ /i V , Ut 5.58 39.3 174.3 19.1 0.02 0.09 0.17 ;*i 1 5 12 j _ 0,25 A 7 7 0, :S 3.03 0.02 5.63 30.6 ! 5? 3 13,7 0,01 1.47 fl fl7 19.3 77 f L 7 142 1 -' 12 2 j 29 0, 2* 0.14 i. vV {)0 C 11 123,5 136. 2 18.5 , 1.02 0.14 * 7 *3 7A 7 1 i A ] 2 12 2 1 0.10 0,15 0.38 0,01 J 4 .-•-) li.L'i 33.1 196.8 24.2 0.01 0,18 0.08 m 144 4 1 i. *•? ^ 2 0.2S 'J ! i / 0.13 3: 43 !: ^3 21.30 435.7 152.3 31.7 0.01 2.70 0,13 LA . i_ 72 9 ' j C i -j ._ : 0.27 0.19 . . 6 3.23 0.02 C 7 C Da /J ii), iii.i ; 22, 9 A " 4 V 1 V i 0.95 0.13 13.7 70 c :^q i 2 12 v 1 0.11 0,15 0.10 -« J 1 * 0.03 10.42 120,7 (71 23,2 0.02 0.18 0.02 , '.47 1 1 9 2 9 f] ^n 0.13 0.18 •; 7-_ . ! i. 0.01 •6.82 Sb.O 117.9 21.2 0.01 1.64 0.14 23.2 7 < 3 [48 [ 2 it 1 3 0.23 0,19 0.15 "1 r: A A £0 6.85 62.7 ; 20 . j 22.4 i 1.38 0.10 -n 72 A 149 1 7 1 "3 1 1 0.10 0.20 . OS 2.51 0.03 4.34 45.9 237.2 19.8 0.02 0.04 0.13 , * 1 *5 1 1 2 0.33 0.28 0.12 3. 4o 0.05 9.33 127.8 170.1 A.* 4 it. i 0.02 1.20 0.18 21.4 73 0" ! r < * i 7 I " 7 _ 0.29 0.27 0, 10 2.93 0.05 5.38 67.6 ISO. 8 a* r 0.00 2.33 0.13 20.6 -}£ 4 152 1 1 _ 13 5 4 0,13 0,23 0.03 ~ j Art A "T V 1 'J / 9.29 96.2 161.6 21.9 0,03 0.59 1 1 10.4 (C^ f i _ j " 5 "> 0.37 (i *"'> J y / j "fp 0.05 6.93 7S.9 155.9 17.2 0.01 2.51 0.17 17.4 70 j 154 1 _ 13 -; ^ 0,35 0.30 Vi i i 3,00 0.04 S.b3 66.9 109.4 * 7 " V V 1 2.00 0.15 19.9 a 1 7 [55 J 7 :: 4 1 3, 32 0.50 0,17 5.14 0.0s S.30 72. S 63.0 24.0 0.01 1.51 „ , 156 t _ 1 -j 1 2 1,31 0.45 q 73 "4 0,04 3.34 147.3 lift "5 7=: " 0,01 1.32 0.42 , 1 c1 2 i j 4 •^ 4 4 c 0.4S r"j ~ 4 .1 i.i 3.79 0,06 8. 38 126.9 ' -• H 31 7 0.00 1 Q " • • A A i C*3 4 f CQ 1 . 13 4 1 i fl T3 V.iJ v * 2 1 0.07 1.49 0, (,j 7.70 230.0 198.2 26.0 0.03 0.S7 0.18 , 159 1 L !: t 7 '. c =: 0.10 1.96 0,03 4,31 101.7 7 1 7 3 23.7 0.00 2.0! 0.28 17.6 55 i 4 ISO | 1 ' -. 1 3 0.48 V * I i 0,10 1.43 0,03 4.13 103.1 226.3 22.8 0.02 1.11 0.26 J c 58 x V; ' 4 i 4 £ 2 I 0.21 0. 12 •"< * j 1 .25 0.10 2.13 -"?P •"; 172.6 13.8 0.01 0.13 0.15 3.5 79 2 i')i 4 j IS ^ 2 /» =7 y . JL 0,13 0.2b ! 7! 1 . / I 0.07 3=05 97.0 530.7 0.02 1 . 59 '} 1 7 19.1 73 f 2C3 t 7 I . . "5 A .4 i fl 1 7 0.24 1,38 0,09 4.53 67.2 196,2 19.2 0.01 0,53 0.13 1 K 31 7 / m 1 d ! J 2 i • . « . i 21 0.08 L.Li 0. 02 3.59 CJ 4 214.3 18,3 0.0! 0,0! fl.03 8.9 7-7 Q 205 4 J 13 2 £ 0.32 0.20 0.15 2.45 A i\c V . V u 7 T 1 . JJ 59.7 169.2 17,4 0.0! 1,43 0.02 17 C : T ? i .-. b 4 g IS 2 ~ 0.20 ''? 0.14 •1 or o.os 5 si 54.5 203.1 19 fl 1 o 7j) 0.03 14.7 74 : 237 4 i 7 4 r 17 2 * 0.11 0.19 0.03 2,19 0.03 4.S5 57.1 Vl7 Q 18.6 0.03 0.04 0.03 3,2 ~ r -\ 2 20a { 3 IS 7 0,33 0.20 0.17 2.41 0.03 7.57 74.1 ICC A iJJ. U 19,7 0.08 0.43 0.27 13.5 " ~; 7 20? 4 IS 2 n 0.18 0.19 ft i A 1.29 0.03 b.55 42.6 217.3 1 R 2 0.02 0.04 0.10 14.2 77 9 210 4 3 16 L 1 0.08 0.13 0.05 1.39 A A -^ 3.04 31.5 010 rj !1 0.02 0.14 0.03 8.0 Q ill 4 3 16 7 2 0.3S 0.20 0,14 2.06 0.04 7.29 76.0 266.1 18.9 U « vi 0.82 0.05 18.0 73 8 7 1 7 f IS 2 xJ 0.24 0.18 0.10 1.86 0.03 5.34 65.7 306.4 18.4 0.02 0.49 A A 7 12.8 70 C J] 5 4 1 3 17 4 1 0.13 0.17 0.07 1.20 0.04 2.81 80.6 330.7 32.8 . Ui 0.31 0.10 9.0 2S4 t 3 1 7 4 i. 0.24 0. 15 A t 4 V. II 1.35 0.03 3.98 63.3 577.7 ! S 1 0.01 1.08 0.10 14.6 SI " 1 '•; I 7 • J 0.31 A 4 Q 0.13 1.09 0.09 3.77 107.9 599.6 21.0 0.04 1.02 0,03 14.0 6! 1 21S J J 17 : 1 0.20 0. IS 0.07 1.42 0.01 5.26 118.7 220.8 20.6 0.05 1.04 0.07 9.5 C7 c 217 1 T 17 1 ': 0.33 0.19 0.11 1.59 0.02 6.50 115.4 Arn a 19,5 0.03 1.09 0.05 16.2 70 i) 218 4 3 • 7 j •J 0.42 0.20 0. 12 1.64 0.02 S.23 180. S 237.5 21.7 0.07 0.78 . 05 14.5 ;1 219 1 J 17 1 i 0,12 o.ie 0,07 1.53 0.01 7.08 66.5 195,5 23.3 0.01 0.57 0.01 9.4 55 -T

PAGE 167

156 OSS ;cA FAS PAD 3PE PAR 2 A 3 m K NA 2u FE m 2N CO "2 :E rp ivono 220 J 2 .7 1 2 0.23 0.23 0.12 1.71 0,03 9.40 90.3 267.2 20 ' 0.05 1.61 0,05 15.4 88,0 i * 4 j [7 1 " 0.30 0.20 0.11 1.50 0.02 6,74 142.1 211.0 2' j . l * * . J ! 0.18 1 A 64,9 122 1 n J r 11 1 1 0,11 0.15 0.07 1.51 0.02 4.23 125.1 228.5 ' a r 0, ; .-3 0.34 0.05 12.3 , 223 I 3 17 i i _ }.27 0.13 0.13 4 77 0.03 3.75 165.6 7 r« 11 7 0.09 i : L~ . 22 17.3 5b, : 224 s q 1 7 1 / 1 3 0.23 0.17 0.10 1.S8 0.02 6.8! 178.7 -Lit J 23, 1 0.05 0.77 0,10 14.7 S4.1 * 3 18 I 2,23 :) ' ^ 0.03 1.35 0.0? 2.61 172.4 3S6.7 7C -7 iJ.Z . 02 0a-2 0.02 9.3 , 226 1 7 18 7 L 0,21 0.23 0,17 2,-5 0.08 5.72 71.8 32 r .5 16,0 U s Jo 0.49 u . 1 1 19,7 7 £,5 7-77 1 ^ p 7 i » 2 1 0.13 ft |0 V , .J•5 «i 0.05 6.17 84,6 323.1 14,6 v 1 y i 0.30 '.i J 15.8 70.3 223 4 3 |g 7 1 ft. ! "3 0.24 O, Iz 2.46 0,14 5.64 72.8 330.2 23.1 0.04 0.12 0.05 ii 7 , 225 «, •j 13 7 2 A ; r U 1 ^ 2) U , . 7 -L . i-_' 0.07 5.31 74.5 133.7 13.3 0,02 0,43 0.04 20,2 77 fi 23C J 18 7 *3 0.22 0.25 0.18 2.48 1 1 62 70.1 290 2 21.9 0.04 0.73 0.03 tq ? 75 A in f 3 ia • 0. 10 •" "7C 0.12 2.33 0.03 9.55 53,5 316:8 ";-* ^ LL 1 0.04 0.14 v« 02 13,6 , i<3i | 3 is 7 ft i e .'. i J 0.18 0.14 2.05 0.04 6.87 / .' . / 210.2 13 7 0.03 A C "t 0.05 19.1 75.5 -'£ i 1 a 7 v 0.2O 0,22 0.16 2.58 0.05 5.35 71.5 245.3 13.4 . 0.01 0.46 0.08 19.5 75,5 234 1 IE 7 * 0.08 0.26 0.08 2,72 0,03 4.67 44.0 284.5 17.8 0.04 0.35 0.01 10.1 , 235 1 3 1 a 7 i' 0.20 U 1 j. j 0.14 2.03 0.03 8.03 73.3 7^0 7 ltd. / 13.8 0.01 0.73 0.03 16.1 78.0 225 1 i ;Q 7 n : 0.13 ft *M '2 2.41 0,02 4.46 52.3 200.3 * O 7 0.01 0.66 0.11 15.3 74.3 23^ | j 19 7 / i A t t 0,28 A ] ? 3,62 0.06 6.72 80.0 248.1 25 5 0.03 0.35 0.22 11.6 , 238 1 n 19 7 i 0.15 0.2* A ic i 2.30 0.05 7.01 155.3 160,3 19.4 0.10 0.37 0.05 23.8 77.4 239 t 19 7 " 0.23 0.25 0,15 2 . 93 0.05 6.22 103.7 171.4 19.5 0.08 0.41 0.04 21.4 75.0 240 1 j 19 / 1 0.03 0.25 0.11 2.93 0.06 6.33 88.8 298.3 30.5 0.06 A 7C. 0.11 12.8 , t j 19 7 ^ 0.23 0.23 0,16 3.06 0.06 8.06 122.0 179.5 20.3 0.06 0.59 0.08 70 1 i.-J i. 77.1 __ i2. 1 a 1 q 7 q 0. 19 0= 20 0,14 2.*3 . 06 6.23 30.3 187.6 0.06 0.57 A i -7 23.9 75.5 Hi 1 3 19 T 1 0.1O U . i y 0.11 2.30 0.11 4.33 47.3 213.1 29.1 Vi iJ . 23 0.10 i 15 v 1 -_' : J . 244 1 2 19 7 i '1 'iC 0.23 0.14 3.01 0.03 8.29 77.6 lJVitl 20.6 0.14 0.79 0.07 23.4 78.3 2^5 1 3 ;3 ] ;, ,22 0.13 0.12 2,53 0.06 5.38 63.0 tlilli 13.1 0.10 0.86 0.11 ">o 9 i.i. 1 i. 74.9 242 1 1 3 19 7 * , , , , . , , , . , a 0.11 16. 3 . 24 7 1 3 19 7 & 0.23 0.23 0.16 4.00 0.05 10.67 97.1 154.1 23.3 0,13 1 1 LJ 0.04 73.2 24E i 3 19 7 ~ 0.15 0. 13 v. 11 2.69 0.04 6.14 116.4 * ft i r iO**. J i J / 0.03 0.54 0,12 24.5 77,1 243 1 3 20 7 J, 0.09 ,22 0.08 2,79 0.04 2.51 43.2 176.0 15.5 0.04 0.50 0.05 3.8 t 250 I 3 20 7 1 Oi dv * ;/ "' V . 1 1 2.91 0.03 8.03 65.3 109.4 14.3 0,09 1.67 0,01 15.7 ^ >; ".-' '' i 1 '; 20 7 / j 0.1b 0.13 0.11 2.14 0.05 16.21 61.5 150.3 i C .« i J.T 0.08 0.76 0.03 15.0 76.4 ? c ; ? 1 ~J t 20 E i 0.17 0.13 0.09 2.83 0.02 5.88 118.1 38.5 Oi R 0.03 0.07 , 11.5 . 253 * 2 22 -; _ 0.18 0,27 ft * * U a Li 2.37 0.03 8.07 30.2 106.2 20.3 0.06 1.39 0.08 5.7 73,0 22i 1 i 20 b j 0.13 0.21 0.10 >•: CI i. JJ 0.03 7.05 60.0 39.4 22. 1 0.07 0.74 0.04 210 72.6 1W i 3 20 £ I , , , , , . , . , t , , 77 £ . 252 ! £U 6 j. fj 1 *5 0.21 0.10 3,00 6.33 £8.3 94,2 16 7 0.82 A A 1 •77 < 1C \ 257 t _ J £ 3 0.23 ft 70 U 1 Li. 3.21 0.03 3.30 75.7 106.2 18.3 0.02 1 ;~!h 0.12 , 77.6 301 t 1 i 1 i I 0.13 0.05 0.06 0.74 0.01 0.44 117.6 207.0 19.4 y.u3 1.03 0.15 5.5 47.9 302 t. i 1 1 J &a 0.06 0.03 0.33 0.01 2.53 107.1 253.0 22.4 0.06 0.41 0.16 3.4 c»3 J w : « J 303 1 i i 1 2 0.16 0.06 0.08 .'1 ifii 0.02 1.41 119.5 -.7= ^ 14.3 0.02 0.46 0.05 6.4 49.4 304 _ 1 1 1 i 1 0.15 0.10 0.06 0,75 0.03 0.79 85.6 213.0 14.8 0.02 0.15 0.06 48.5 305 9 1 i 1 4 A 0.32 0.11 0.08 1.01 0.01 1.19 136.5 252.4 j 5 Q 0.02 0.38 0.13 5.3 50.5 205 £ 1 1 ; 2 0.27 0.11 0.07 0.49 0.04 1.08 212.1 210.0 14.2 0.03 2.77 0.05 6.0 51 •} 307 n 1 1 1 | i 0.17 0.10 0.05 0.64 0.02 0.59 38.3 280.1 1 A ' ivi i 0.02 0.29 0.07 4.6 44.8 308 5 1 1 1 0.30 o.io 0.06 0.58 0.02 1.65 1 -7A 1 306.6 14. 7 0.04 1.78 0.19 8.0 54.5 303 2 1 i < i 1 0.20 0.03 0.05 0.53 0.03 0.80 82.9 233. 1 10.7 0.03 1.57 0.09 5.0 49.3 310 2 j 1 1 1 0,22 :,13 0.06 0.63 0.02 1.54 242.3 375.1 25.4 0.10 8.27 0.04 3.1 . 311 I 1 2. 0.35 0.13 0.10 1.53 0.04 2.70 261.1 403.5 31.4 0.67 0.65 0.02 13.5 . 212 2 1 1 1 <5 0.3S 0.15 0.09 0.75 0.04 1.74 245.1 375.2 26.5 0.06 0.18 0.16 13.5 39.3 3 J _ 1 £ i ! 0.29 0.09 1 '-' -2* . 1 wO 0.05 1.36 81.3 308.3 18.5 0.02 0.23 0.01 6.4 47,4

PAGE 168

157 OBS SEA FAR PAD SPE FA? CA ^b NA Ca t 6 0.11 -:"> i •"> 0.05 0.97 0.01 4.32 34.7 126.2 27,1 0.02 0.19 , 7,5 52,7 320 2 j £ 1 2 ii.27 J , >.} f 0.10 0.89 0.02 1.70 104.0 < 7S ft i / J V Li: '') 0.04 0.32 0.15 t -. .-, z~ j1 9 1 i 3 0.28 0.03 0.10 0.96 i).02 132.1 162.6 25.8 0.02 ft ~>u 0.01 9 3 62,5 V ' i i 7 i 2 0.15 0.17 0.09 1.95 0.03 3.00 120.1 93.3 13.7 0.02 0.30 0.01 11.7 , £ » -• I 2 0.44 0.16 0.14 2.72 0.03 5,15 234.3 112.7 16.3 0.03 0.42 0.12 4 q n 57," 324 2 1 t 3 0.36 . 1 7 0.15 9 ^Q 0.03 3.13 298.9 114.3 23.0 0,01 0.30 V. ij 15,3 54.3 325 o i 7 i 4 j.ii 0.1S 0.07 t =,9 i a J£ 0.04 4.39 129.5 100.2 26.3 0.04 0.35 0.04 19 :. i J I V o2, E 326 t i J _ 0.2b 0.14 0.11 1.56 0.05 3.82 233.3 154.5 21.0 0.05 0.36 0.10 20.0 6^.7 327 2 1 7 J 3 0.16 0.20 Oi Ob 1.05 0,02 1,68 58.0 52.2 11.1 0.02 0.20 0.07 4 n c iO. J 61.9 328 2 4 1 -Z i ]*"' * '" 8 '" ;C 0.11 1.95 03 0,32 102.3 168.2 99 A 0.04 0.34 0.05 10.5 , tj a it j 2 1 9 2 0.52 0.23 0.23 2.50 0.03 2.55 133.3 183.4 15.6 0.00 0.34 0.01 13.4 7"3 1 330 ' 9 1 3 v 0.40 0.26 0.21 2. 52 0.02 3.49 180.6 205.7 17.3 0.01 0.36 0.01 19.7 53.8 331 2 1 1 3 1 i 4 1 0.17 0.16 0.10 1.19 0.02 0.84 131.7 215.3 15,9 0.01 0.48 0.05 8,3 54.7 33i 2 J B i £ 0.08 0.04 U . ; .'i 0.39 0.00 0.83 37.0 53.4 2.6 0.02 0.27 0.04 17.1 68,2 i JO £ 1 3 1 9 0.37 0.17 0.14 1.39 0.04 3.02 137.1 309.3 29.0 0.02 0.30 0.05 13.1 57,2 234 9 i 3 9 1 0.07 A 0.06 1.50 0.01 0.77 67.7 7 •) 1 L 0.01 0.43 0.11 7.8 , -r 2 1 1 i3 i 2 0.40 0.32 0.15 2.73 0.03 2.41 32.7 31.1 9.5 0.02 1.16 0.10 15.8 67.9 336 ! q 2 9 0.24 0.12 V 4 1 1.73 0.01 1.20 268.3 121.3 20.7 0.01 0.59 0.01 14.0 59.3 :37 2 1 3 i | 0.19 0.13 0.10 1.19 0.03 1.33 59.2 149.8 13.8 0.01 0.28 0.01 5.7 44.9 ;36 2 1 i 2 0.37 0.24 0.14 1.71 ft M 2.25 101.1 145.5 2? 2 0.04 0.41 0.03 12.3 58,8 333 •> 1 3 i 3 0.27 0,21 0.13 1.70 0.04 1.31 78.8 156.0 20.7 0.05 0.23 0.10 10.2 54.4 340 9 1 3 2 ! ft 1 9 v Lii 0.08 1.91 0.03 1.26 47.8 145.9 8.1 0.04 0.26 0.05 7.2 61.1 341 £ 1 9 J 2 2 0.34 0.26 0.13 9 in £ -jj 0.02 4.35 68.8 4 9fl c i£3i J 12.3 0.01 0.18 0.14 15.3 68.4 342 iy 1 3 i 3 0.34 0.24 0.12 2.15 0.03 2.92 80.3 146.5 12.1 0.02 0.49 0.18 12.6 53.9 344 9 i i 3 4 i 4.12 0.28 0.27 2 50 0.07 5.74 191.9 86.1 21.9 0.05 0.03 0.07 , , 345 1 1 3 4 3 1.58 0. 22 0.20 * 9 9 £ £0 0.06 4.80 126.9 119.9 14.5 0.02 0.29 0.04 21.0 53.2 346 2 1 1 1 0.13 ft '••) Jm ll 0.08 1.66 0.03 5.42 55.1 146.1 14.2 0.02 0.44 0.01 8,9 51.3 347 i 1 : 1 i 0.42 0.19 0.12 1.57 0.02 2.76 232.4 126.1 i ~ i i J. i 0.02 1.09 , , . 343 9 1 4 i 9 0.32 0.25 0.12 2.09 0.02 4.93 83.0 172.9 17.9 . 04 0.38 0.09 11.0 56.5 349 9 1 3 ^ 1 0.13 0.31 0.21 2.13 0.02 7 91 55.7 156.6 29.7 0.35 0.11 0.02 ; . : ft jJv 2 i 3 9 2 0.37 0.23 0.23 2.34 0.03 1.11 106.5 146.1 Ll ,0 0.03 0.22 0.06 15.4 sn .4 3S1 n 1 'i 9 2 0.35 0.24 0.22 1.31 0.03 5.41 124.8 150.8 36.6 0.03 0.19 0.07 8.5 51.8 "" i \ 3 i ii 0.45 0.24 0.16 7 f)1 J . V J 0.04 7.q5 118.7 nec 9 £JJ.£ 20.4 0.07 0.13 0.02 22.3 1L ") . » 4. 354 i 1 i 2 3 0.48 0.24 0.16 2.36 0.03 3.25 94.0 242.7 W "7 U . / 1 0.36 0.04 4 n j ta B 4 55.3 ;C^ 2 1 ~ ^ i 0.20 0.22 0.07 1.55 0.04 1.14 132.0 176.1 * c 7 0.02 0.74 , , . £ 1 3 9 £ 0.33 0.20 0.14 2.38 0.03 5.50 87.5 173.0 i 1 "> 0.04 0.57 0.01 17,1 72.3 :-7 2 | 3 2 3 0.35 0.22 1.93 0.03 2.44 143.1 156.9 4 n r ll.J 0.03 0.61 0.02 13.5 63.7 358 2 1 1 4 £* 1 0.13 0.13 0.07 1.10 0.03 1.77 47.2 179.9 8.3 03 0.25 0.05 c 9 55.3 359 I 4 2 2 0.45 0.17 A 4 "i 1.56 0.03 0.65 373.0 275.4 ! 1 1" tin J 0.06 0.84 0.05 , 350 9 £ ; 4 -i £ 3 0.26 0.17 0.09 1.32 0.04 0.85 103.2 181.9 10.7 0.02 0.25 0« il 3.1 ^9 9 331 > J 1 4 1 I i 0.12 0.13 0.07 1.05 0.03 1.58 53.2 182.0 13.5 0.03 0.22 0.08 7.0 49.7 3E2 i i 4 | 2 0.57 0.17 0.18 1.34 0.03 3.16 113.4 280.9 21.5 0.06 1.04 0.15 12.2 52.3 363 2 l 4 1 3 0.42 0.14 0.13 1.15 Vi j2 • *7C 36.0 239.4 21.0 0.04 0.65 0.17 8.5 55.3 :g4 £ 1 .» 4 2 < 0.08 0.14 0.07 1.27 0.03 0.89 47.8 130.9 7.6 0.03 0.21 0.03 5.3 L 4 "T 3l. .' -I. 7 l 4 2 i. 0.29 0.19 0.14 1.97 0.03 2.60 38.9 120.3 6.2 0.03 0.30 0.11 14.2 65, 1 366 9 £ i 4 ^ 3 0.10 0.10 0.07 0.88 0.02 0.85 36.8 109.3 4.3 0.01 0.20 0.23 10.7 50,4

PAGE 169

158 3BS 3£A FAR PAD SPE FAR 3A p *3 K NA 01 "m it CQ ,*,n 2E :_-1V0HD 2b7 2 , 4 i 0,15 0.28 0.14 3.17 0.03 5.10 37 "! J t m U 143.1 43.0 0.07 0.11 0,13 , . 3bo ^ i 4 j _ 0.35 0.24 0.17 2.50 0.03 5.22 105.4 121.3 27.8 0,10 0, 16 0.20 13 s 61,9 369 7 i 4 n n 0.27 0,26 0.17 2.28 0.02 7.85 100.1 118.6 21.3 0.06 0.18 0.05 11.8 59.5 370 2 1 4 i 1 i 5.13 0.15 ft A 7 1,70 04 LlUt 59.7 f CI ™ i J i i ^ j T 0.22 0.12 0.16 1.54 0.01 3.51 54.7 125.3 6.4 0.06 0.11 ft 17 10.0 50.4 384 4 t 2 7 0.27 0.13 0,21 1.46 0,02 4.72 C7 ft J / . V 150.9 14.6 0.17 0.10 0.05 9.5 49.6 385 2 i / j i 0.11 0.14 0.08 * 01 0.04 1.08 45.2 352.5 17.1 0.04 0.22 0.06 5.0 53.5 386 ^ i -7 0.54 0.17 0.20 1,57 0.03 1.84 86.9 524.5 0.07 0.54 0.03 10.1 63.5 337 £ 1 o i 0.21 0.14 0,11 1.21 0.03 1.17 97.7 436.3 9.1 0.03 0.25 0.04 7.7 55.8 389 2 1 2 2 i. 0.26 0.08 0.18 4 -:>' 1 . L'-J 0.01 3.30 45.8 163.5 8.5 0.16 0.10 0.21 9.1 46.9 390 1 j " 0.41 0.23 1,72 0.02 3.93 ~-3« L 260.8 15.6 0.05 0.16 0.55 9.3 50.6 401 _^ 7 . ; : 7 2 0.42 !* 0.12 3.11 0,08 8.99 133.4 352.7 27.3 0.04 0.34 0.06 27.7 58.0 402 v 2 13 7 V ft 7 * 0,25 0.09 i 52 0.05 6.83 110.0 374.5 23.0 0.05 1.34 0.10 "" *J 52. 1 404 7 i " 7 7 0.42 0,23 0.09 1.82 0.07 4.55 115.3 380,4 24.2 0.05 1.57 0.09 22.6 62.3 405 > 13 7 0.35 0.33 0,09 2.07 0.05 5.77 123.9 330.5 ">0 *3 0.02 1.13 0.05 18.4 60.2 407 2 7 i ~ 7 £ 0.51 Oj L.£ 0.09 1.88 0.12 3. 52 507.1 ^ " : ! 7 22 3 0.04 1.93 0.17 21.5 56.0 403 ^ £ 13 7 '• . 37 0.21 0.03 1.33 0. 10 4.55 343.1 241.0 46.2 0.03 1.23 0.12 22.7 33.0 410 ^ 7 13 7 1 n„"B 0.25 0.11 u , y J 0.06 5.10 155.6 243.0 29.4 0.02 2.04 0.08 27.4 56.2 411 __ £ 4 n * J 7 'J 0.33 0.25 :), ;/ 3.42 0.07 7.01 159.7 "J 7 '" ; c , 33.3 0.03 1 wU 0.10 29.3 "7 * "7 412 • h 1 0.19 . l § O.OB 1.98 0.27 7.91 232.3 381.1 38.5 0.05 7 77 . ! < f i i. 1 . 413 2 7 1 *5 s 3 0.26 0.24 0.10 L,. Lxl 0.15 7.72 131.4 343.1 22.9 0.04 2.78 0.09 22.5 . 414 ^ 4 i j G 2 0.36 D . 1 9 0.08 2.63 0.13 7,00 155.0 251.7 24.2 0.01 2.37 !) 77 y . Li, 21.6 . 415 7 14 4 • i 0.08 0.04 0.04 0.55 0.01 2.00 90.3 315.6 ";" C . 0.04 0.32 0.09 5.8 . 416 ^ 7 14 4 i ^ 0.24 V 1 i ft ' 2 4 <)4 0.03 3.71 153.4 598,6 23.5 0.04 0.71 0.06 17 -/ 49.3 i * 7 £ _ 14 ; j 0,22 0. 12 0.10 0.33 0.01 1.41 123.6 526.1 i-Ji 3 0.04 0,07 0.11 10.3 A A .': 419 £ L 14 1 4 A i i 0.06 A A5 0.50 0.02 3.14 103.4 338.8 22.5 0.03 0.56 0.05 5.3 . W £ L * 4 a 0.19 0.08 0.08 0.32 0.03 2.35 185.3 417.2 21.4 0.03 0.33 0.0! 10.4 AC D i *:4 o £ 14 •1 4 1 0.11 0.07 0.05 0.57 0.02 2.55 85.4 424.1 23.8 0.02 0.61 . . . 423 2 14 1 3 0.26 0.10 0.12 0.39 0.01 5.00 109.7 540.3 26.9 0.03 0.75 0.11 , . 424 fc 2 6 4 1 i 0.09 0.09 0.05 0.86 0.02 2.13 104.0 251.9 rJWtl 0.04 0.09 0.02 7.0 46.4 425 J _ b 1 L 0.25 0.14 0.09 1.14 0.02 0.96 293.7 709.7 34.8 0.05 0.48 0.13 . . 425 L 2 G J «3 0.20 0.11 0.08 1.12 0.03 2.05 229.2 516.8 35.4 a An U.Dj 0.32 0.14 3.5 49.7 428 7 6 2 2 0.15 ft 4 4 0.05 1.27 0.02 4.05 93.0 245.7 13.0 0.02 0.40 0.02 11.3 65.2 429 ^ 6 T. 0.16 0,09 0.05 1.14 ?\ •" 100.4 244.6 12.6 0.02 0.08 0.07 11.0 56.1 431 2 7 ^ £ 0.27 0.13 0.09 1.35 0.03 3.35 159.1 495.5 20.7 0.04 1.03 0.13 12.6 bi .j 432 A £ 7 6 £ n 0.24 fi 13 0.08 1.26 0.02 1.30 150.0 418.3 29.5 0.03 0.68 0.22 11.6 54.7 433 ^ t 15 i 0.14 0.17 0.04 1.67 0.04 3.15 53.7 258.9 25.3 0.04 0.35 0.09 . . 434 L 15 " ^ 0.26 y. ;/ 0.07 1.93 0.03 3.50 98.3 293.8 iDi w 0.01 0.40 0.03 * 7 7 55.7 435 7 / ;5 2 3 0.19 n 4 n y . 1 3 0.02 1.39 0.05 3.33 73.3 296.3 17.4 0.02 0.34 0.09 (7 7 H.M i. 70.0 43b 2 o 15 2 1 0.09 0.14 0.04 1.67 0.02 3.01 47.9 £31 24.8 0.05 0.38 0.04 3 , 2 .

PAGE 170

159 ." j SEA C 1 ^ PAD --jPAR CA J *3 i NA cu ^ n :n 22 M0 BE DP 2V0-D -2~ ^ c 15 2 0.22 0.16 2.09 0.02 5,23 91,2 233.0 17,0 V . V i 0.40 0.07 13.3 / J . J 439 2 2 15 2 3 0,13 0.16 fl ?*' ^ ' 1~! 0.01 n en j J i bJ, 3 206,1 20,6 M ;•;" 0.40 0,04 10,5 _"7 7 439 2 > l ~ i i 0.15*) 0.12 0.03 0, 99 0.02 1 ~56 274. 1 23,9 A AC 0.38 9. 25 5.2 c Q t" I "; r t Q ' i 0.13 0.03 1-01 (;„ 02 * y _ 7Q * / J . 7 407.5 23.1 M . v5 A A " , i4j 2 2 '^ l" J '"• 1 *5 A t "3 0.04 0.01 1 7! 3* 1 ! 23.4 0.04 ),3£ '\ '} ~7 2\4 442 } 2 . J i 1 ' ;". 7 1 2.C3 1 . 05 0.02 ^ . a i i. J l 7 220.5 1 3 * : . . 1 i 0.45 J , J 1 b. u ~i -_ . i*3 ~) 2 1 i 0.15 A 1 2 r* 1.30 0' 1^7-K 282.4 23.4 > j": 1 :, z ^ 444 2 2 4\ c 1 J 0.13 V i ii 0.05 1.17 'J . y L i.Dt .til J 3-6.2 26. 2 0.04 ?) ." 1 A 7.9 •4 \ r 2; I 7 _: 7 4 0.06 [ ,04 v. vO } , i'i 3 0' j ". ~ 121.7 219.8 8. 1 ': 3 ' 0.09 A. 1 A 3, _ -.v . *, i ;|} 7 -" 0,40 " 1 7 O i-7 i "A •ftj 7 77 1 ' Q '! 657,5 -, -. . ft A *A A 7 ( l -.. J '.' i _ * 1 ' ' £ '' * * -* ' .' « A -j i i JV .' , V * i.i w / 1 iJ.il _i , i V V Z .' : j ,' >!, iB . i.. / 502 2 19 7 jj 29 J. ll 0.11 1 * C 0,01 1,74 105.4 508.4 47.5 0.02 1.24 y , i.u ft C 2 \ 503 2 T J 19 7 / 1 0.07 0.04 0,03 A ^ J V. it ft A \ 1.37 138.2 256.1 3.9 0.06 0.72 0.08 4.1 504 2 A '; H 7 0.2B :• t * i. y > I i 1,23 0.02 1.38 32.9 504.5 17.4 0.02 v.bD v', :9 t ( 4 ll SAC 2 1 J 13 7 g CIS 0. )9 0.07 0.75 0.02 1.56 127.3 394.6 11. 1 0,03 "C V ( tw A •:-! -• . i 1 i ~ n : 5 2 J 19 7 * 0.12 0.05 0.04 0.24 0.01 2.49 181,3 231.5 t r a 1 b . U 0,04 0.74 0,03 5.1 7 ' g 507 2 j 13 7 2 C.33 n * >-. 0.14 2.01 y , y j *Ii « j 176.3 475.0 21,6 0.01 0.19 20.2 2' n 509 2 _ 19 7 i 0. 21 o.so 0,08 l . y j 0.01 2.06 152.0 299,7 17.9 0.02 0,30 0.08 15.5 m\ 5 509 2 a 19 7 1 0.13 0.05 0.04 0.25 0.01 1.49 136.4 239.2 12.8 0.04 0.38 . c o J. £ 510 L n 19 7 2 0.30 f r\ 3.11 1.44 A ^"3 2.45 135,8 554.5 14.2 0.02 0,35 0.08 '2 2 Z r H r ! ' 2 9 19 7 *5 0.20 0.03 0.07 0.56 0.02 1.21 113.0 375.1 16.1 0.01 0.20 0.05 13.0 47 c 5;3 2 *j 2i 1 2 036 On lG 0.05 I i 29 0.05 2.61 224.1 507.0 20.0 0.C1 1,05 0.08 12.8 C" \ 514 2 a 23 1 3 0.40 0.15 f\ * fl 1.20 0.07 2.19 237.9 592.3 21 3 0.01 1.55 0.07 10.6 51 ,4 — SIS 3 23 i 2 0.45 0.18 0,12 1.50 0.05 1.85 123.5 467.5 16.7 0.04 \ t n 0.07 11.9 r 1 3 C t ~ 2 3 23 i 3 0.44 0,19 0. il i . 32 0.07 2.17 141.7 7 3 j , 7 20.1 0.05 2.02 0.05 11.3 2* 22; 6 519 2 3 00 4 i 0.49 ;. 4 7 0.18 S.47 0.11 5.01 126.1 828.6 28.8 0.02 1.18 0.07 13.5 r 5 3 r 1 .2 "; 23 1 3 5.61 0.19 0. '5 1 , 40 0.13 7.17 153.8 713,3 33.2 0,03 1.37 0.01 r j 2 J T 22 1 _ . 25 J iii 0oS9 1.48 0.02 3.35 171.2 372,7 13.1 0.02 a nn 0.07 10.5 u 3 R 523 > 3 23 1 ^ 2.23 0.11 0.09 1.47 0.02 5 OS £.. i. j 205,4 372,3 15.1 0,02 0.46 0.05 10.4 rr 2 524 2 -. 7» LI I 0. 12 ittijl 0.25 1.13 0.03 * nq iii., J 228.4 13,8 0.04 35 a a c 3. i 525 L 3 -' _ 2 0.35 rt * 1 .: 10 1.90 0.01 •J. 20 139.0 10.3 0.02 0.33 0.07 12,5 "" 5 : k ; 2 j 21 2 ? 0.12 U 1 *.' 0.06 0.83 MS 0.70 102,0 248.6 10.0 0.01 0.28 0.13 6,2 :r 2 523 £ j " t 2 _ ft TO V .17 0.09 1.33 0.07 3.33 143.7 239.1 ' »1 i 0,05 0.75 02 10.5 r. r 529 *. 7 <--4 2 •5 0.28 0.14 0.08 1.29 0.03 1.97 110.3 298.0 I> t 0.01 0.42 0.0? 7,8 -7 4 7 A •"5 2?1 £ i 0.08 v * * 7 1.00 0.03 1.33 81,7 194.3 10.9 0,04 0.10 0.02 r r J J i 2 J *--4 2 I 1.34 0.15 0.13 1.36 0.10 2.63 185.5 292.2 14.3 i) c . 0.69 , i i " 532 i n 21 2 7 0.26 0.15 0.09 1.20 0.08 73.0 274.3 9.7 A A A 0.08 0.07 3.3 51 I; ":!'.'; 2 1 7* i 4 i ' i 0,10 0.05 0.49 0.02 2.39 126.2 229,5 a. 5 0.04 0,30 0. 09 4.5 _ 534 2 "i) 0)1 i 2 1 i*t 0,12 0.12 0.82 0.04 107.8 342.3 . 2,05 32 0.03 9,2 j 2 r "*c; 2 n li i 3 •j. iC 3.11 0.07 0.75 A At 2.49 217.0 285.3 8,7 0.23 0.29 0.04 , i :3 c J JO 9 3 * 7 1 i i 0.16 0.03 0.06 0.52 0.02 6.70 222.7 301.1 ! 7 3 0.06 0.48 0.11 5.4 _ JO / £ ~ | 7 I 1 i 0.28 '"> 1 Q 0.10 V. U J 0.03 2.24 577 -. 464.3 15.7 0.07 0.94 AT 5.5 49 7 538 £ 3 17 i j 0.28 0.10 0.03 M. w„ j . 2.01 213.4 435.8 i 5. J ft fl "3 0.56 0.02 7 c — -T G 533 £ n 17 1 i i 0.15 0.10 0.05 0.81 0.01 2.48 154.7 248.0 19.2 0.06 a nn A A, 7 3 . J 540 2 w 17 i i > 0.32 u. II 0.09 0.95 0.01 7 .-c 207.5 366.3 22.3 i j , y /; 0.45 0.05 i u . V 35 541 3 17 | d 0.26 .'. i i 0.07 1.02 0.02 2,38 153.6 432.2 24.4 0.04 0.62 0.06 3.0 :: Q 542 •> J $ 7 { 0.13 y 1 1 1 0.04 0.75 0.01 0.55 64.0 315.1 19.2 0.05 0.27 0.04 6.0 49 1 543 2 j 1 / 4 2 r \ 32 0.13 0.03 0.94 0.02 1.26 217.0 463.0 23.8 0.04 0.85 0.02 1 1 i --A 1 544 2 3 • 7 1 3 0.23 0. 13 0.08 0.96 0.02 1 . 39 153.4 426.0 24.2 0.06 0.75 0.07 11.6 p 545 17 1 4 0.18 0.09 0.06 0.50 0.03 1.33 107.5 iijt i '16.7 .A ,A A 0.72 0.09 6. 2 44 Q 546 > 3 17 | 9 0.32 0.10 0.08 0.49 o 02 1.53 32.2 358.8 17.5 0.06 0.48 0.02 10.4 547 i d 17 4 i ;,0.41 0.16 0.11 0.77 0.03 173.5 405.7 20.8 0.05 0.85 0.07 9.3 3 2 -

PAGE 171

160 r -5 ::.--.r 3 C : ~z 3 A^ ; AS K ;; : :: RK IN CO jig :: ^3 rvotffi 2 3" ' . '; c .V: i.-i ).73 5.0! t.39 :0.0 206.fi :-.! j jg " '0 • Jfl , -5.0 : : :: ; : ,: : ;'o? !.:£ .01 £.10 It"'" 2 1 ".: ::': J.07 ' 58 5.5S3 0,00 '. '. 7 S0.4 58.3 ~. , '.'. -Z '' ' ;'_ J , .: ... " "* , : .:.! '.' , .4 '.;? . . 5 :. 2 50.1 ~ * .".' :,-, ?. 03 '..;-; 119.1 -12. 9 24, 2 ;, ' i '.:: ; . : o 1' i 55 ; ; ;; . . ! 3 ., .? :.:; :.;i :2:.: 225,2 25:, 4 ...4 ),4* :!.-; 58.9 _ .. '•' • 4 L7; £.2: 7 .' . 5 lEi.S :.: ).02 ; ": 46.3 : 3 ;; | :,25 '; ' 1 .is f _ IB : J3 J 4. .4 75,5 2:4,7 2. 7 . 33 0.05 0.45 . :s ::.: 53.4 • :: . )S ' . 75 :.j] 39, " 337.4 3. ^2 5, 2*j . 53 6.7 -3.2 " " " ; . :J . ,: .. i2 : .. ;i=0.2 «S1.4 5.3 j. 06 C>i5fi }. 03 J 3 « : . ._ ).2fl ;.io ,:-4 ;.-)! :.3! "5.4 556.4 7.1 0,05 . 52 1.09 3.2 72.: •'

PAGE 172

BLOOD SERUM DATA CODES 161 OBS SEASON 1 2 FARM 1 2 3 CLASS 1 2 3 NUMBER CA MG P ZN FE CU SE = Observation number = Rainy (1987) = Dry (1988) = San Jorge = Don Benito = San Francisco = Pregnant-lactating ewe = Lamb = Yearling = Animal number = Serum calcium, mg/dl = Serum magnesium, mg/dl = Serum phosphorus, mg/dl = Serum zinc, ppm = Serum iron, ppm = Serum copper, ppm = Serum selenium, ppm

PAGE 173

IUNBES .c :,?; 162 .3 2.32 0.15 23 14 89 20 3! 3B 33 40 41 42 43 43 4E 47 48 43 50
PAGE 174

3.9 :.\9 163 30 ; 7 :45 :; '° 4 ' l'.7i :.'•! 4>5 >'« ;'!" 173 :,3 ; i' : .w " ' "'" 77 : ; 130 11.4 2.'c9 S23 li'Jn •'M? 5 ' 143 •1, ,, , ," -'" '•!•'' 0.17 0.050 :s . ; M 19.2 1.34 8.SS o.63 2 .35 1-7 »,» 63 ,' : ; :: = ;.I «35 2.71 O.SC 5.14 3.51 £ j| 57 sa ; & .ft lil '•" ;" "4 0.73 0.207 3 535 w." : ' 3 '« J"2 ^ °' fil 5 37i U, ',! ;•}' =" : 75 :« 0.69 o.:n7 ? 1 7" v .;', :;' ,|37 «» i.a mo 0.029 ?: ; ::; ;?? 13 4 --' >•« ;,;: : . M ,.« n \ n : -7. 80 : 1 : 2:5 '!' j? J' 57 : '' 83 '43 5-39 C047 * : ; ,;;', :'":' ;-' 4 °' 33 !•* 1.27 0.014 — " '->.a i.« 4,53 0,33 ' "J i ,-,a n .,,, : ;44 15.7 1.89 '3 ) » J « „ . 3 3 '49 •-, 4 " "~ " "" 5 i-M 0.064 *" 'J. 9 £.20 2.85 t.3| ; 54 t m ,,, i 47 2.00 .3 '55 1 19 1 mi 036 '0 5 7 77 7 ,, . . " M "' z '-OH i g j a a ;s ;:,•: 3 a 31 i -3 3S9 : o'' i« r« •' r3 2,£ M1 °'°« 7 3 :•: ' '« C ' 77 '* «.M 0.0O7 ... •'" '7 * 4.43 0.70 9.61 j jo , «„ : -' 11.3 2 35 e 77 -, ., ... '••'' ,._. t, J E *« y.81 £.63 -go n -,dc 3 557 ::.) 5 77 ^ r 7 , ., . " "" J ' vUi! 1 665 in a -, ! J,J ' J ''* 0.014 1 »" iO.S I.93 7 47 no, , „,, , 3 379 .. , ,, !'™ °S " -<•' l.H 0.343 32 : 34 ! 3 57 ; 33 : S3 i 50 1 3 3 ~, •' L Jj 3 '-3 0.78 2.5J 55 ft <«, 1 f 3 ' £ : ' 73 X:4 ».7B 2.52 0.86 « 3916 '" J' I 6 "2 ; ' 1S 3 '» 'J/ »» : . ;b 0.34 0,035 93 : 94 2 35 35 : 37 ; 33 : 33 2 oo : I >

PAGE 175

'.a 5.043 ,:30 154 123 ;:4 126 !27 i:9 13S ill 139 HO 141 .4a :47 :48 :49 150 . =i*J •' '':'< 4.41 5.57 1.33 1.15 1.024 t:3 »•! :."5 4.6} },73 1.77 5.35 2.006 :1b ".0 1.38 J.33 ).71 2.70 0.35 0.022 : 73S7 : "3 "X33 4.33 5,41 3.13 0.36 0.043 =3 3.40 2,:: 0.54 2.02 0.35 0.018 ' '' -'32 1.39 0.49 2.59 1112 0.076 -•3 2.33 2.34 0.65 1.33 1.20 0.049 , ! 5 7 2.20 3.43 J.32 1.64 0.35 0.070 2 . _ 7.0 3.02 3.74 0,76 1.86 0.30 0.023 J °91j] 5.8 3.73 4.30 0.78 1.43 I. 01 0,081 3 7859 3 734; £•3 2.3a 3.21 0.48 3. Id 1.12 0.024 '•1 2.1: 3.27 0,35 1.33 0.36 0.070 ; 35! 2.5 2.77 4.76 0.41 1.35 1.31 0.012 3 7233 3 7417 -' 2.41 3.90 3.73 5.0 2.22 4.37 :.24 1.17 1.32 O.130 1.13 0.070 3 7333 6.5 2.40 3.79 ;,7.1 1.23 1.13 0.076 : 136 6-3 2.12 2.42 1.03 1.40 0.139 * :5 5.3 2.12 3.03 0.49 2.59 1.22 0.155 1 :i ''• 1.70 5.30 0,35 1, 35 1.14 0.117 ' 1170 6.0 1.32 3.36 0.39 2.43 1.21 0.132 3 1 6.4 2.03 5.33 0.66 1.55 1. 22 0, 072 4 7-0 2.00 '.55 0.54 : 35 0.33 0.244 * ' j i« 3* 2,65 'J, u3 2.67 1 . 24 3 132 3 155 3.0 2.00 7.44 0.71 5.4 2.05 4.23 0.32 15a 1 :i J.31 0.055 1.07 0.278 3.73 0.072 » y t . 'j J . 1 S 0.0a 1.21 0,106 3 701 5.1 2,07 5.20 0.30 2.31 V34 0.1OS =. J 2.29 3.50 0.20 .39 .03 j.113 3 15 5.5 2.33 3.30 0.71 .79 .58 0.071 3 173 5.1 2.47 4.17 0.71 .o2 .38 0.103 3 134 2.5 2.25 3.32 0. : i . 11 .25 3.339 S 130 5.4 2.23 4.06 0.33 .23 .20 0.059 132 3 147 4 137 3.3 2.10 4.32 0.59 2 6-2 1.54 4.02 0.83 1 5.4 1.90 4,17 0.64 2 .07 .23 1 .10 : .43 :. 232 47 0.124 .33 0.034 3 315 5-5 2.13 2.65 0.80 ! 54 : .12 0.133 3 115 6-0 2.17 3.12 0.52 1 77 : 33 0.08] 473 3-3 1.70 4.37 0.59 2 12 36 1.044

PAGE 176

343 Hf.H 't.JSS 4j J.::' 165 174 !8C .33 uo m IS2 :S3 :J4 :58 195 •24 5.0 1.43 i.s 35 . 4 2. 55 :.*D i 2.0/ :.3 50 „ u 45 7.0 2.1! l,o; 6,4 :,; : ' :,:: IS s.4 !.SS 1.2* :a 7 -j -jii ii j_ i 10.4 1.;: S, 24 f 3.6 I.8C ?< 33 4 '.6 [ f 3 5.47 j 7.0 1.13 5.24 1.45 3.7 I* (3 S.4 :.62 1.34 242 5,7 2.3b ? ai 715 5.0 1 52 3,4' 457 5.0 1.53 2. -J :3d = ^ 1.31 • =233 5*7 1.34 2.23 4,2 " A i •itVi *31 i , )'3 7 77 -.3 :.:i ^.57 04 4 3 2. ^7 " 53 '" -._. .._; .'.vu5 ..-5 2.1* 1*06 3.323 3. SO 3.32 :.4i 3.352 ;.4i 3.35 :.53 3.323 3.74 3.25 i.7i 3.053 0.S3 :.74 0.33 0.043 'j.zd "^ 1. jl 0,353 : . oo 4.15 :.43 0.054 3.76 2.32 1.25 0.059 5.57 3.44 2. .'5 0.006 1.26 1.99 3.72 0.027 ..2! 5. 40 1.12 0.043 "-. : J . . 13 . i '.-•'. .37 3.62 1.13 0.013 .52 3. 39 0.54 0.086 2.23 1 , 04 0.01! .23 2.27 0.31 0. 054 4.90 3,56 0.141 .32 3.53 1.17 0.022 • SO 1.73 1.73 3.065 .65 5. 12 1.45 0.054 1.12 ,055 5.3 33 3.34 758 3.3 2.22 3. 15 0.S1 0.75 303 E.i 1.34 3,35 0.50 773 5.8 2,01 3.94 0.72 303 £.5 1.80 5.04 0.52 7 25 5.9 1.98 ".43 0.21 741 5.5 1.66 4.45 0.40 747 5.0 2.56 4,22 0.61 1.15 .076 1.34 3. 353 1.-5 0.040 1.37 0.041 1.13 0.053 , :, : ; 0.070 1. "' 0.070 1.68 0.053 1,03 0.054 1.58 0.053 1.13 0.114 1.39 0.053

PAGE 177

166 MiF

PAGE 178

167 BONE DATA CODES OBS SEASON FARM CLASS NUMBER ASH CA P MG = Observation number = Rainy (1987) = Dry (1988) = San Jorge = Don Benito = San Francisco = Pregnant-lactating ewe = Lamb = Yearling = Animal number = Bone ash, % dry, fat-free basis = Bone calcium, % dry, fat-free basis = Bone phosphorus, % dry, fat-free basis = Bone magnesium, % dry, fat-free basis

PAGE 179

166 40 41 41 43 44 45 43 30 73 53.3 ;;• 33.4 24.34 20.30 22.16 22.36 21.41 2^.30 721 35,4 22.27 -:, i 21.73 911 60.5 20.54 3:2 32.6 22.77 313 52.7 22. 13 322 35.2 23.03 336 51.3 22.22 359 jit 21.42 2673 54.3 24.49 2346 51.4 22.92 l.X 3.43 7.41 5.77 3.37 5.2E 7.54 7.00 a. 37 B.S9 3i24 3.33 ),4C 5.44 ;.40 :,53 :.40 3.55 3.33 0.23 0.23 0.36 3.30 0. 31 ).45 .41 0.43 0.44 0.35 ).38 0.43

PAGE 180

R8E3 -:a 169 1 -231 55.3 14.:: i «!0 ;>.: :j.'oi 3.N 74 T 5 7 6 73 73 Si 30 31 92 33 34 33 56 37 33 39 :oo :t~ -::.17.53 3.3/ _ 4] ihSl 21.42 7.77 j.55 53:7 -::-.; 21,51 -_:; ), 40 £581 32.3 22.22 3.31 0,39 '343 £2.3 '/ 22.54 3.J7 0.33 "172 50. 7 22.13 2.45 3. 33 ":o 31.5 22. )£ 3,35 1.33 7558 52.3 23.74 8.54 0.30 35146 33.7 22,22 2.34 0.44 76145 il.i f* -"7 3.54 3.38 76154 37.7 20. 34 3,53 0.35 7C1S 30.2 ::.7i 7, 13 1,-1 73 53.2 26.31 10.25 0.33 35 5!.: 52.43 3. 57 0,i3 115 54,7 2' -3 3.07 0.-2 !SS 73.5 ^0. 59 13.47 0.96 137 o4. 5 12.20 7. JO 0.4! ::s S4.fi 23.20 :. -5 0.22 .1! 54.3 22.20 ".31 i.37 54.3 22.17 :. 24 . 42 56.0 23.05 i.i* !.3S " ,c 65.1 22.51 i . 74 0.44 225 btf.b 15.54 3.40 0.32 ::3 34.4 -.« t 7 22 1 ., .,-3 134 55.3 22. 3a 5.74 0.25 us 52.1 1 3 . 51 5.23 242 53.4 21.33 ;.. : 3 0.27 316 55.4 20.59 3,33 0.41 33! 33.2 19.46 4.33 .' . 43 356 23. 3 29.33 '.34 0.39 157 60.5 :o.45 5.53 0.43 359 31,7 19.16 g t 39 0.42 3 '2 60.7 ?t 27 3.51 0,42 421 54.6 22.46 6.3! 5.43 437 65.2 21.53 7.37 0.37 473 52.3 22.20 3.30 •.43 504 53.3 22.20 4,32 0.46 608 61.5 -1.13 3.47 3.55 b!0 51.3 -. ., o,g2 0.43

PAGE 181

"23: '-'iI; :S,B 170 134 :23 140 141 !42 :48 m 30.3 21.24 7,57 0.41 7"~ r _ , * . , j 3 :. 44 5,39 . 73 51. 3 15.57 £.53 :' 0.42 ESS 25, 5 20.43 3.30 J. 43 336 :z i 22.03 7.23 '.'. 2i i;oc 52.5 22.36 ",72 ).44 I51£ 53,6 21.24 7 0"* 0.30 -;40 33.7 -4.i.b 7.05 3.32 -Jib Oii2 22. 34 7.53 0.35 5S06 55.4 22.34 7. SI '.31 5310 :o.O 22. 4^ :: '.36 5360 £5.3 21.60 £.54 5952 £5,2 21.33 7.21 0.27 ?""" 54.3 22.14 7.52 3.25 '417 55.3 20.72 «. JO 7-2' £4.3 22.54 ' = ^37 "357 55.1 22.00 3.££ 0.36 7891 £2.9 22.15 :,30 fl IJ* 7:51 52.9 17.38 7.23 :,54 "67 43.0 15.45 3.14 o,2 5 7:71 52.3 13.11 4,33 ; , 31 "377 52.6 13 " c 2.21 0.21 41.6 14,20 4.94 '' . 2 7 73S3 i 3.3 17.23 1,57 0^22 7334 43.4 13.30 4.41 0.22 7332 51. ; 13.17 3.32 0.23 mu }7, £ 22.19 7.33 0.34 23170 St. 3 22,32 ".17 0.35 33142 £5.3 24.13 3.35 0.33 73131 £3,3 22.63 75251 £5.0 23.47 0.41 73143 ££.£ )g 53 4.70 K34 73163 ;e c 22.55 0,53 73203 51.4 2 1 50 2.07 0,31 7320! 40,4 14.44 4.10 0.23 7S264 43. 4 16.53 3.25 73273 41.4 13.72 4.55 0.25 "3100 -6.3 15.34 3.33 0.20

PAGE 182

171 WHOLE BLOOD DATA CODES OBS = Observation number SEASON 1 2 = Rainy (1987) Dry (1988) FARM 1 2 = San Jorge Don Benito 3 = San Francisco CLASS 1 2 = Pregnant-lactating ewe Lamb 3 = Yearling NUMBER = Animal number HTO = Hematocrit, % HB = Hemoglobin, g/dl LEUCO = Leucocytes, /ul NEUT Neutrophils (mature) , % LYM = Lymphocite, % MONO = Monocyte , % EOS = Eosinophil, % BAS = Basophil, %

PAGE 183

172 U83 rfta ka *n & AS NUflB 1TQ -3 LEiffiO LYfl SON 203 3AS I 1 1 4942 35 ;:.5 21700 72 2 1 1 520 30 3.7 17500 7! 5 3 1 2346 20 10. 1 10760 85 4 3632 32 10.9 22300 33 2 5 i 5632 23 10.7 16800 37 1 b 1 2220 24 11.3 15100 68 7 1 426: j! "' « 20400 38 1 1 2678 30 10.2 20600 66 1 15 9 5340 23 10.2 11351 72 2 !0 1 4*610 35 12.3 30500 70 2 7 j il J 3316 23 10.5 21200 84 3 12 1 ! 1 7658 29 3.6 9910 77 2 ;3 1 1 2 7830 37 12.3 14560 79 1 14 1 1 2 7272 32 10.3 16600 96 15 ! 1 2 73134. 39 12.9 24300 93 2 16 1 1 2 76145 3! 10.3 13200 30 1 17 1 1 2 731 13 37 12.5 22000 83 1 1 ia 1 1 2 78199 36 11.9 18100 52 1 1 IS 1 1 2 7960 31 10.9 24000 93 1 20 1 1 2 78142 35 11.2 15500 79 1 21 ; i 2 78111 34 11.3 11350 97 22 ! 1 i 73131 35 11.8 10329 87 j 1 1 2 7620 34 11.2 15300 30 1 24 1 2 76154 35 11.6 12590 93 1 25 I 2 7243 35 11.0 20600 32 3 J 26 1 3 6233 40 12.1 7854 93 1 27 1 3 5413 30 11.2 200O0 87 3 20 1 3 5883 34 12.3 M200 32 29 1 3 6895 37 11.6 11344 37 1 30 j 7 6831 39 12.0 15000 86 31 1 1 3 6417 34 11.4 9950 9! 3 32 1 i 3 6309 38 11.3 9824 95 33 1 1 3 6867 35 11.4 17300 82 34 1 1 3 6881 34 11.5 10334 79 5 35 1 1 3 5229 26 11.3 11553 35 2 } 36 1 3 S313 38 13.1 24100 82 1 5 37 1 2 1 22 44 14.4 8700 68 2 2 33 1 2 1 132 39 13.0 5000 76 20 39 ! 2 1 38 37 12.3 8200 63 1 5 40 1 2 1 146 40 13.3 7400 75 1 8 41 1 2 1 106 37 12.3 6800 73 1 42 1 i 1 110 46 15.3 7700 66 2 2 43 1 2 1 261 35 11.6 9600 76 1 44 1 2 1 60 44 14.5 7800 63 1 7 45 1 2 1 102 35 11.6 5900 70 4 7 46 1 2 1 236 43 14.3 6000 79 1 1 1 47 1 2 1 78 42 14.0 8000 58 48 1 2 2 212 42 14.0 3400 92 2 49 1 2 2 2 43 14.4 7500 78 2 1 50 1 2 2 194 43 14.3 3100 33 2 1

PAGE 184

033 SEAS F«8« CAS SJBB «T0 '-EUC0 LVD BON SOS 3AS 173 SI 1 2 2 173 48 :i.O 14900 77 ; 8 52 1 2 2 76 41 13.5 65O0 36 3 1 53 1 2 2 73 38 12.6 £000 38 54 1 2 2 130 <2 13.9 3800 33 2 55 I 2 2 114 33 11.5 5000 37 4 2 •) 56 1 2 2 536 44 14.6 BMC 96 i CT 1 2 2 112 45 14.9 ":-:; 57 4 1 ) 55 ! 2 1 504 45 15.1 6500 30 2 59 ! 2 3 131 45 15.0 7100 76 3 60 I 2 3 49 32 10.6 7500 57 : 10 6! 1 2 3 113 40 13.3 S350 81 : 62 *l 2 3 107 39 13.0 S500 73 j 5 S3 : 2 3 111 38 12.6 5300 47 5 64 : 2 3 759 41 13.6 3000 85 3 65 1 2 3 125 39 13.0 iZHO S3 1 1 66 : 2 3 127 39 12. J 10000 75 3 67 1 2 3 135 40 13.3 7700 31 2 68 ! 2 3 121 46 15.1 6500 90 69 I 2 1 312 41 13,4 6650 S3 2 5 70 ; 2 1 225 40 12.2 7400 52 11 71 1 3 1 282 42 14.0 6350 57 9 72 i 2 1 277 44 14.6 10750 62 9 7 3 1 3 ! 936 il 13.5 7500 60 1 12 74 : 3 1 295 40 13.3 7600 93 3 7 75 ! 3 1 244 41 13.6 7700 32 10 76 1 3 1 922 44 14.8 9600 54 g 77 1 3 1 731 45 15.1 8000 72 ') 6 78 1 J 1 313 41 13.5 9400 71 3 9 79 1 3 ! 286 41 13.6 3100 73 3 3 80 1 3 3 959 41 13.5 8500 60 2 ;7 8! 1 2 3 226 46 15.2 7800 62 12 82 1 3 3 339 34 11.3 9900 74 5 6 83 1 3 3 311 36 11.7 6500 SO 3 84 : 2 3 606 39 12.9 9500 58 3 12 35 1 3 3 637 38 12.6 S800 64 5 56 ! 3 3 217 40 13.3 6100 67 2 37 I 3 3 657 37 12.3 10200 66 1 S3 ! 3 3 249 35 11.6 9300 62 3 89 : 205 41 13.6 7000 52 11 30 2 1 2 7818 39 14.5 7550 30 1 31 2 1 3 7831 36 12.8 6440 71 3 I 92 2 1 3 7357 39 12.7 6440 66 1 93 2 1 3 7891 32 12.7 6800 45 8 94 2 1 3 78143 36 12.9 6100 47 1 6 1 35 2 3 78151 30 10.0 5650 59 : 4 1 36 2 3 73163 36 10.9 5150 53 2 ! 97 : 3 78203 38 12.2 9100 63 1 7 1 98 2 2 78205 42 14.5 9250 91 99 2 2 78243 42 13.6 7250 85 2 • 00 2 2 78264 29 13.8 7950 70 3

PAGE 185

174 DBS SEAS WH CLAS NUI1B HTD LEUCD LV» ra« EOS CDS 101 102 103 104 •05 106 107 108 105 110 111 112 113 114 115 116 117 113 119 120 121 122 123 124 125 126 127 128 123 130 131 132 133 134 135 . 136 137 138 133 140 141 142 143 144 ; 145 ; 146 ; 147 2 148 2 143 2 150 2 3 78273 7 G 23 "962 7567 7971 7377 7993 7984 7932 79100 5500 5910 5960 76131 75201 7417 48214 7233 2940 5E170 3916 58142 1915 626 5963 37 37 36 26 40 35 41 J3 36 35 27 23 36 37 11.7 11.0 13.2 13.1 9.1 13.7 13.5 13.0 14.4 10550 9500 7300 '-10 5250 3500 6B 2 3 3 4 15 24 54 78 80 35 93 106 115 42 33 34 29 35 34 34 45 42 34 126 47 137 33 2 3 147 150 151 152 :54 163 170 1B6 132 215 10.3 9.6 12. 1 12.2 9.3 7.3 13.0 10.9 10.0 10.1 7.5 13.1 11.7 3.5 13.1 13.4 12.1 12.7 11.3 13.5 11.3 11.1 12.6 16.0 14.9 11.4 15.9 11.1 14.2 12. B 10.5 12.8 13.3 10.2 11.0 12.2 13.8 13.6 5350 5750 7050 3900 7250 3850 30 6950 70 9550 83 3350 30 6650 53 4500 32 53 ' 7250 3650 78 11200 39 6500 74 5200 73 4650 73 7800 34 10750 66 3400 73 6000 78 9650 62 5250 3" 1 7200 60 7700 61 5650 77 6450 68 3500 35 3950 30 3900 9 12 51 2 5700 68 4900 73 5000 77 4250 97 4150 73 4650 67 7050 73 8750 56 5000 32 3300 97 9500 72 3500 74 9000 10300

PAGE 186

175 OSS SEAS FARM CIAS NUNB HTQ KB LEUCO LYN hon EOS 5*3 151 2 7 3 473 34 12.1 3500 67 2 3 \ 152 2 2 3 701 37 12.3 5250 57 3 Q 1 153 2 2 3 7C2 36 12.4 3050 51 7 1 154 2 2 1 1170 35 12.7 6950 53 7 1 1 155 i 3 2 ; 47 16.6 3950 46 5 1 156 1 3 2 3 47 16.1 3400 43 3 1 2 157 2 3 7 5 40 13.6 2000 74 158 2 3 2 6 41 13.7 5400 14 3 159 2 3 1 137 34 11.3 4300 57 2 1 160 2 3 3 225 43 13.7 5000 51 10 161 2 3 1 226 40 13.8 3100 44 12 162 2 3 1 233 44 15.0 6500 54 t 11 163 9 3 1 234 23 9.8 3200 73 1 4 164 2 3 1 236 34 3.0 1800 32 6 165 2 3 1 242 36 12.3 6000 71 5 1 166 2 3 1 246 33 13.5 4600 93 167 "i 3 3 216 41 '.3.8 4600 75 ) 1 168 t 3 3 331 36 13.2 4100 82 3 163 2 3 2 555 37 11.3 3400 70 1 170 2 3 3 357 37 13.4 5500 74 1 17! 2 3 3 358 41 14.8 4500 31 2 1 172 2 3 3 372 41 14.0 3550 40 2 2 1 173 2 3 3 375 36 11.5 6500 84 1 174 2 3 1 421 35 11.8 5400 76 1 6 \ 175 3 3 1 434 42 17.0 5; 00 65 6 176 2 3 1 457 41 12.3 3400 51 7 5 177 2 3 3 604 41 13.5 5200 80 4 1 173 i 3 3 608 31 11.1 4500 33 I 179 2 3 3 615 41 14.0 4900 78 1 1 1B0 2 3 3 649 38 13.3 4300 71 1 11 2 181 2 3 3 7!5 42 14.7 4400 74 1 3 182 2 3 3 716 36 11.3 3700 74 183 9 3 3 725 42 12.9 4800 56 5 184 2 3 3 741 42 13.4 4550 98 5 1B5 2 3 3 747 40 :2.8 3300 65 1 136 2 3 3 758 37 13.3 4200 34 1 1S7 2 3 3 765 44 14.5 3500 82 188 2 3 3 769 33 13.2 3550 73 4 189 2 3 3 772 41 13.6 3000 48 130 2 3 3 777 44 13.5 4350 83 1 191 2 3 3 773 40 12.7 4400 54 192 2 3 3 809 31 10.7 5000 83 2 193 2 3 1 906 39 12.3 5050 69 2 194 2 3 3 5000 46 14.3 3900 70 2

PAGE 187

LIVER DATA CODES 176 OBS SEASON FARM CLASS NUMBER MN ZN FE CU CO MO SE = Observation number = Rainy (1987) = Dry (1988) = San Jorge = Don Benito = San Francisco = Pregnantrlactating ewe = Lamb = Yearling = Animal number = Liver manganese, ppm dry matter = Liver zinc, ppm dry matter = Liver iron, ppm dry matter = Liver copper, ppm dry matter = Liver cobalt, ppm dry matter = Liver molybdenum, ppm dry matter = Liver selenium, ppm dry matter

PAGE 188

27 29 40 41 42 43 44 45 45 47 43 49 50 34.3 . i9 11. J .-5.4 •34 3.9 25.2 215 J.3J 75.0 21? 11. 4 35.5 .25 U.3 5:. 7 22S 12. 1 H.S 226 13, 1 55.2 244 3.3 72.3 2-: . .-. c 55.4 255 '. ; 115. S 332 ;7, ] ::5.3 333 7.4 57.3 "36 14.0 50,0 506 1 3. 5 532.7 11.2 77.6 731 12.6 107.3 ill 12.2 33.3 313 3.2 58.3 522 11.5 "3.5 325 10.7 55. S 559 12.2 52.4 2220 3.3 "4.3 2673 3.7 73.3 2345 5.5 57.7 3522 :.2 130.0 4310 ',2 50.3 5222 3.2 55.: :533 10,2 125.4 5'2!5 7.3 59,2 5233 5.5 173.0 5417 9.2 149.5 .'.7 7.7 '.24.2 5331 3.0 113.7 5331 3.0 137.0 7652 3.5 72.0 75154 2.3 33.0 73134 £.2 72.3 73145 3.3 42.5 177 F; 32 32 "0 3E -i -, . ,-. -*...173 J . 5243 1:1. ; 25. fl 0.302 1 . 535 316.5 102,: 0.2:: ;,434 : -.2 ::.3 :,3:3 ;.'.$'< 329 4 15.4 0.264 :.22: . 53.5 5.055 .322 ?3» a 25.2 0.155 3.S57 . '.z'.'.Z 6". J 3.235 :.; : . --i 54.9 0.224 3.667 . 223.4 15.7 0.230 5.545 . 520/5, 12.1 0.334 0.375 . SOS, I 13.4 0.210 0.331 . 425.5 25.5 0.335 j.393 . . c . . •h? 1 ' ftM C7t ..J. 1 .-. .BJ .'. j/i , 233.5 121.4 0.121 0.J39 . 33.5 62.5 8.143 0.054 . '.35.5 75.3 0.034 0.515 . 545.5 54.6 0.265 0.520 533.5 43.3 0.233 0.521 . 252.4 131.2 0.223 0.5O5 . 2::.; 23.6 0.112 0.271 . 504.3 122.4 0.7B3 0.523 . 163.6 108.2 0.090 0.202 . 452.2 276.6 0.162 0.619 . ..., j 30.2 0.080 0.355 . 1 7 . * 4 58.5 3.133 0.517 . 475.5 21.7 0.156 0.530 . 501.2 91.0 0.149 0.533 . 237.3 37.1 0.095 0.36C . 3:4,2 33.5 0.255 0.434 . 433.3 111.7 0.112 0.456 . 241.0 :E6.7 0.132 0.502 . 231.5 24.0 0.264 i.564 64.1 33.1 0.275 0.222 . 450.3 23.7 0.237 0,477 . rrg • :05.2 0.325 0.634 . 270.3 21.: 0.035 3.159 . 281,6 S3O.0 23.4 0.443 0.403 . 265.3 104.5 1.337 0.692 , 535.2 20.2 0.365 0.719 . :52.£ 108.3 0.193 0.554 . 540.2 2-. 7 0.140 0.351 . 693.5 22.4 0.254 0.632 . 533.4 33.0 0.232 0.640 . 2:5.4 64.8 0.332 0.472 . 132.5 33.1 0.065 2.530 . )32 7 50.1 0.205 0.209 . 64.4 .4.5 0.172 0.245 .

PAGE 189

178 73 EO SO 91 54 3: 56 37 39 100 : :-.j 1 33. 5 555. 3 5.2.3 3,169 ..1:7 .2.2 1.4 35.3 :l, 5 55.2 3.334 j7 ::.: 31.5 551. 5 7 3.044 2.114 t 3 232 3." 133,7 : 35, : 55. ;.i23 1.151 5 MA 51.5 ::55.3 254,5 ' .l-i 1. 15: 5 331 3.2 545.5 22. 1 1.55; ,.521 )&l 333 ...4 -55.5 554, ; 50.5 3. 1 12 l.;24 1 333 .55.5 5:5.2 252.5 1.025 1.024 -. _ 2~ 524 5,7 113.4 524.0 54.5 0.026 0.123 333 \i. ' 54.1 205.4 226.2 1.013 .027 :.74 3-3 ".: : 3 . . 5 173.2 145.5 0.031 3.034 . , S3 2 3:3 /.5 1 5 1 . 2 I""-. " lll.C 0.023 0.030 3 331 3.2 -5 -' 553.3 131.6 ;•, 0.023 ) '' 2 III 7.3 77,3 174.4 "5.5 1.015 0.003 ':." 5: 7 :.S 35,0 553.2 103.3 0.040 0.013 :.3fi 3 325 53.5 275.5 2:6.3 0.023 0.033 \. .z 37: 3. ; 1:5.5 ^i j 155.5 5.052 1 .225 '.. 22 3 2~5 9,4 •5, 1 515.3 121.2 0.025 2.223 t.40 ! 431* 7 ' 106.1 330.3 53.7 0.054 2.034 j\ -:" ". 3 55.5 229.6 132.5 0.034 0.027 1.02 3 473 ". : . 3 37.S 555.: 25.3 0.155 0.235 i 33 4 212.5 '17 3 0.038 0.022 1.22 3 5)8 ij.i 140,7 :33:.3 £23.3 2.021 0.03S 0.75 3 oio 7,0 73,7 330.3 143.3 0.023 2.025 3 515 3.0 53.3 100.3 47.3 3.033 0.006 2.30 j 5-5 3.5 53.7 54-J.2 "5.3 2.031 0.021 1.23 3 733 : ,3 33.5 -54,4 71. ! 0.034 2 ,013 3 ~15 ;2.3 ic ' 206.1 1:7.3 ... 332 1.027 1.45 3 7'6 ".4 12-4.5 511.2 155.5 0.022 0.521 2.14 717 3. 3 53.7 552.7 50. 3 '. 529 0.020 1,02 t 725 3.3 55.5 227.5 15.0 .. ,5_ 0.025 1,53 3 741 ;. 4 55.5 275.4 133.5 5. 525 0.028 2.30 3 747 11.4 54.3 221. 5 :51.2 -.025 0.025 3 hS ".7 56.5 212.2 :24.2 0.125 0.225 0.72 3 "35 •? 4 35.3 415.5 :14.5 1.134 0.011 2 . 54 7 S9 :2.o 55.6 555. 1 1 25 53 . 22 0.055 0.326 1.4: 772 10.2 33,3 331.7 57.6 3.060 3.329 1.33 T " 7 S.2 37.2 254.2 110.4 1.024 1.123 1.05 3 773 33. 3 505.4 :22.7 1.023 2.023 0.94 3 90S 533 • < 1 79.0 26.7 55.2 0.051 O.011 1.51 . :: 3 lOOO 3.3 54.0 370.4 53.1 5.016 2 ; .014 5.78 1 1315 13.3 55.3 220.8 "5." 0.203 O.051 1 5340 1.01 1 3*15 0,32 22.3 1506.2 0.022

PAGE 190

EAS FARM :U33 MB HH ZN 179 3 "357 2 33! * 7329 5*125 2 3337 2 3339 43I1-; 23170 I ; 3:40 3 '3:31 2 30.3 506.2 21. 1 1.143 0.303 133.5 1:43.2 .-?.; 3,029 :.;os 133.7 :37.4 /9.6 3,035 :.003 33.3 32 3." '. ': . 1 1.040 3.003 31.3 -15. 1 IS.; 0.072 1.031 -4. 1 377.3 54.9 j f 35 0.150 33. J :33.; IB. 3 0.063 0,031 3:, 2 289. 1 : 4 " . s 0.153 0.003 31.2 S58.3 45.3 o. i ;o 0,043 33.3 121.9 IE 9 'M23 3.013 33. 3 343.4 27.3 0.03b 5.031 22.4 3.354 8.004 1.060 0.315

PAGE 191

LITERATURE CITED Apgar, J., and J. A. Fitzgerald. 1985. Effect on the ewe and lambs of low zinc intake throughout pregnancy. J. Anim Sci 60:1530-1538. Arnold, G.W., W,R. McManus and I.G. Bush. 1966. Studies in the wool production of grazing sheep. 5. Observations on teeth wear and carry-over effects. Aust. J. Exp. Agric Anim. Husb. 6:101-109. Bahia, V.G. 1978. Techniques of soil sampling and analysis. In: Latin American symposium on mineral nutrition research with grazing ruminants, pp 27-29. J.H. Conrad and L.R. McDowell (Ed.). Department of Animal Science, University of Florida. Gainesville. Black, C.A. 1967. Methods of soil analysis. Part II. Am. Soc. Agron. Madison. Wisconsin. Black, C.A. 1968. Soil-Plant Relationships. 2nd Ed. John Wiley and Sons. New York. Black, H.E., c.c. Capen, J.T. Yarrington and G.N. Rowland. 1973. Effect of a high calcium prepartal diet on calcium homeostatic mechanisms in thyroid glands, bone, and intestine of cows. Lab. Invest. 29:437-448. Boling, J. A., T.O. Okolo, N. Gay and N.W. Bradley. 1979. Effect of magnesium and energy supplementation on blood constituents of fall-calving beef cows. J. Anim. Sci. 481209-1215. Bradford, G.E. and H.A. Fitzhugh. 1983. Productivity of hair sheep and opportunities for improvement. In: H.A. Fitzhugh and G.E. Bradford (Ed.). Hair Sheep of Western Africa and the Americas: A Genetic Resource for the Tropics, pp 17-18 Westview Press. Boulder. Colorado. Brady, N.C. 1974. The Nature and Properties of Soils. (8 th Ed.). Macmillan Publishing Co., Inc. New York. Breland, H.L. 1976. Memorandum to Florida extension specialists and county extension directors. Gainesville: IFAS Soil Science Lab. Univ. of Florida. Brenner, I., and A.H. Knight. 1970. The complexes of zinc, copper and manganese present in ryegrass. Br. J. Nutr 24:279-285. 180

PAGE 192

181 Caja Agraria. 1985. La Caja Agraria ante el sector rural colombiano. Inv 148-151. Bogota — -«j« <»3i. U i..i.u auvc ex t»etjtor rural ??o°?rJ' an0 ; I nventarios del sector rural. Cap 1 Tomo 2 pp Cleef, A.M. 1980. La vegetacion del paramo neo-tropical y sus lazos australo-antarticos. Colombia Geografica. 7:9-10. Cohen R.D.H. 1980. Phosphorus in rangeland ruminant nutrition: A review. Livestock Prod. Sci. 7:25-32. Cohen, R.D.H. 1987. Supplementation practices of grazing liyestock-Macrominerals. in: Proceedings, Grazing Livestock Nutrition Conference, pp 93-99. Jackson. Wyoming. Combs, O.K. 1987 Hair analysis as an indicator of mineral status of livestock. J. Anim. Sci. 1753-1758. Committee on Mineral Nutrition (CMN) . 1973. Tracing and treating mineral disorders in dairy cattle. Centre for Agricultural Publishing and Documentation. Wageningen. Netherlands. Conrad J H. 1978. Soil, plant and animal tissue as predictors of the mineral status of ruminants. In: J.H. Conrad and L. J:^™ 1 =?•>• PP 143-148. Latin American Symposium on Mineral Nutrition Research with Grazing Ruminants. University of Florida. Gainesville. Conrad, J.H. , J.c. Sousa, M.O. Mendez, W.G. Blue and LR McDowell. 1980. Iron, manganese, sodium and zinc interrelationships in a tropical soil, plant and animal system. In: L.S. Verde and A. Fernandez (Ed.) Fourth World Conference on Animal Production, pp 38-53. Buenos Aires Argentina . Cooper M. 1968. A comparison of five methods for determining the sulphur status of New Zealand soils. Trans 9 Int Congr. Soil Sci. 2:263-271. Dahnke W C and EH. Vasey. 1973. Testing Soils for Nitrogen. An^lv^; cm *"? J -°Beat ° n (Ed ) Soil Testing and Plant Analysis. Soil Science Society of America, Inc. pp 102-106. Madison. Wisconsin. Dick, A.T., D.W. Dewey and J.M. Gawthorne. 1975 Thiomolybdates and the copper-molybdenum-sulphur interaction in ruminant nutrition. J. Agric. Sci. (Camb.) 85:567-568. ^"'cficlum^nH 11 LUCaS 1973 Tes ting Soils for potassium, «2 » L a ? mai 3nesium. In: L. M. Walsh and J. D. Beaton (Ed.). Soil Testing and Plant Analysis, soil Science Society of America, Inc. pp 133-135. Madison. Wisconsin

PAGE 193

182 Egan, A.R. 1980. The assessment of nutritional requirements of grazing beef cattle. Aust. Meat Res. Comm. Reviews 40:1-6. Ferguson, J. A., H.J. Anzola and R. Pastrana. 1987. Algunas caracteristicas del suelo y de la vegetacion de un paramo de Cundinamarca. Carta Agraria. No 282 pp 3-8. Bogota. Fick, K.R. , L.R. McDowell, P.H. Miles, N.S. Wilkinson, J.D. Funk and J.H. Conrad. 1979. Methods of mineral analysis for plant and animal tissues. (2 nd Ed.). Department of Animal Science, University of Florida. Gainesville. Fontenot, J. p. "1982. Hypomagnesemia in ruminants, in: J.C. Woodard and M. Bruss (Ed.). Comparative Aspects of Nutritional and Metabolic Diseases, pp 95-120. CRC Press Inc. Boca Raton. Florida. ' Foth, H.D. and B.G. Ellis. 1988. Soil Fertility. John Wiley and Sons. New York. Foy, CD. 1974. Effects of aluminum on plant growth. In: E.W. Carson (Ed.). The plant root and its environment, pp 601642. University Press of Virginia. Charlottesville. Gallaher, R N. , CO. Weldon and J.G. Futral. 1975. An aluminum block digester for plant and soil analysis. Soil Sci. Soc Amer. Proc. 39:803-806. Gammon, N. 1976. Plant Nutrient Deficiency Symptoms. IFAS Cooperative Extension Service Circ. 435. Univ. of Florida Gainesville. ' Gentry, R.P., W.J. Miller, D.G. Pugh, M.W. Neathery and J.B. Byrum. 1978. Effects of feeding high magnesium to young dairy calves. J. Dairy Sci. 61:1750-1754. Giduck, S.A. and J. P. Fontenot. 1987. Utilization of magnesium and other macrominerals in sheep supplemented with different readily-fermentable carbohydrates. J. Anim. Sci. 65:16671673 • Goodrich, R.D. and W.R. Thompson. 1981. Sulfur. Anim. Nutr. Health. 38:24-30. G °° ne fo = S e ' !£;!," L 3 ^ 1 ^ R -KChaplin and D.A. Christensen. 1989. Profiles of cu in blood, bile, urine and faeces from Cu-primed lambs: effect of " Mo-labelled tetrathiomolybdate on the metabolism of recently stored tissue cu. Br. J. of Nutrition. 61:355-371. Grace, N.D. 1983. The mineral requirements of grazing ruminants . The New Zealand Soc. Anim. Prod., Occasional Pub. 9. Palmerston North. New Zealand.

PAGE 194

183 Grace, N.D. 1988. Recent developments in trace elements in animal production (invited review) . Proc. Aust. Soc. Anim. Prod. 17:42-46. Gross, C.F. and G.A. Jung. 1981. Season, temperature, soil, pH, and magnesium fertilizer effects on herbage calcium and phosphorus levels and ratios of grasses and legumes. Agron. J. 73:629-634. Guhl, E. 1982. Los paramos circundantes de la Sabana de Bogota. Litografia Arco. Bogota. , Hambleton, L.G. 1977. Semiautomated method for simultaneous determination of phosphorus, calcium and crude protein in animal feeds. J.A.O.A.C. 60:845-852. Hansard, S.L. 1983. Microminerals for ruminant animals. Nutrition Abstracts and Reviews-Series B. 53 No 1 Commonwealth Bureau of Nutrition, pp 24-35. Harris, W.D. and P. Popat. 1954. Determination of the phosphorus content of lipids. J. Am. Oil Chem. Soc. 31:124127. Hartley, W.J. and A.B. Grant. 1961. A review of selenium responsive diseases of New Zealand livestock. Fed. Proc 20:679-688. Hidiroglou, M. and J.E. Knipfel. 1981. Maternal-fetal relationships of copper, manganese and sulfur in ruminants. A review. J. Dairy Sci. 64:1637-1647. Hodgson, J.F. 1963. Chemistry of the micronutrient elements in soils. Adv. Agron. 15:119-159. Holman, J.J. 1944. Studies on the hematology of sheep. 1. The blood picture of healthy sheep. J. Comp. Pathol. 54:26-31. Humphries, W.R. , M. Phillippo, B.W. Young and I. Bremner. 1983. The influence of dietary iron and molybdenum on copper metabolism in calves. Br. J. Nutr. 49:77-86. Hurley, W.L. and R.M. Doane. 1989. Recent developments in the roles of vitamins and minerals in reproduction. J. Dairy Sci. 72:784-804. Instituto Colombiano Agropecuario (ICA) . 1980. Planif icacion de la Investigacion en el Sector Agropecuario. Instituto Colombiano Agropecuario. Bogota. Instituto Geografico Agustin Codazzi (IGAC) . 1986. Atlas Basico de Colombia. Centro de Informacion Geografica. Instituto Geografico Agustin Codazzi. Bogota.

PAGE 195

184 Jackson, M.L. 1958. Soil Chemical Analysis. Prentice Hall Inc. Englewood Cliffs New York. Jacobson, D.R., R.w. Hemken, F.S. Button and R.H. Hatton. 1972. Mineral nutrition, calcium, phosphorus, magnesium and potassium interrelationships. J. Dairy Sci. 55:935-944. Kamprath, E.J. 1972. Potential detrimental effects from liming highly weathered soils to neutrality. Proc. Soil Crop Sci. Soc. Fla. 31:200-203. Kincaid, R.L. and J.D. Cronrath. 1983. Amounts and distribution of minerals in Washington forages. J. Dairy Sci. 66:821-824. Kincaid, R.L. , J.K. Hillers and J.D. Cronrath. 1981. Calcium and phosphorus supplementation of rations for lactating cows. J. Dairy Sci. 64:754-758. King, J.O.L. 1971. Nutrition and fertility in dairy cows. Vet. Rec. 89:320-328. Knebush, C.F., J.L. Valdes, L.R. McDowell and J.H. Conrad. 1986. Macromineral status and supplementation of grazing steers under tropical conditions in Guatemala. Nutrition Reports International. 33:917-929. Lakin, H. W. 1972. Selenium accumulation in soils and its absorption by plants and animals. J. Anim. Sci. 1712-1726. Langlands, J. P., j.e. Bowles, G.E. Donald, A.J. Smith and D.R. Paul. 1981. Copper status of sheep grazing pastures fertilized with sulfur and molybdenum. Aust. J. Agr. Res. 32:479-485. Laredo, M.A. and H.J. Anzola. 1986. Concentracion mineral y nutricional en las diferentes fracciones de la planta. Revista Acovez. 35:4-12. Bogota. Laredo, M.A. and G. Martinez. 1984. Variacion del contenido mineral del pasto Puntero ( Hyparrhemia rufa (Nees) Stapf) bajo pastoreo en zona de ladera. Revista ICA. 19:131-140. Laredo, M.A. , R. Pastrana and H.J. Anzola. 1989. Fluctuaciones minerales en pastos de clima frio. III. Falsa poa (Holcus lanatus ) en lluvia y sequia. Revista ICA. Bogota (In press) . Latteur, J.P. 1962. Cobalt deficiencies and sub-deficiencies in ruminants. Center D> Information du Cobalt. Brussels.

PAGE 196

185 Lebdosoeko j o , S., C.B. Ammerman, N.S. Raun, J. Gomez and R.C. Littel. 1980. Mineral nutrition of beef cattle grazing native pastures on the eastern plains of Colombia. J. Anim. Sci. 15:1249-1260. Lesperance, A.L. , V.R. Bohman and J.E. Oldfield. 1985. Interaction of molybdenum, sulfate and alfalfa in the bovine. J. Anim. Sci. 60:791-802. Lindsay, W.L. 1972. Inorganic phase equilibria of micronutrients in soils. In: J.J. Mortvedt, P.M. Giordiano and W.L. Lindsay (Ed.) Micronutrients in Agriculture, pp 41-57. Soil Sci. Am., Inc. Madison. Wisconsin. Little, D.A. 1972. Bone biopsy in. cattle and sheep for studies of phosphorus status. Australian Veterinary Journal. 48:668j 670. Littledike, E.T. and J. Goff. 1987. Interactions of calcium, phosphorus, magnesium and vitamin D that influence their status in domestic meat animals. J. Anim. Sci. 65:17271743. Loneragan, J.F. 1975. The availability and absorption of trace elements in soil-plant systems and their relation to elements in plants. In: D. J. Nicholas and A. R. Egan (Ed.) Trace elements in soil-plant-animal systems, pp 109-139. Academic Press, Inc., New York. Mack, S.D. and R. Pastrana. 1987. El pasto Falsa poa (Holcus lanatus) , su uso y valor en los paramos de Colombia. Carta Agraria. No 282. pp 15-20. Mayland, H.F., T.R. Kramer and W.T. Johnson. 1987. Trace elements in the nutrition and inmunological response of grazing livestock. In: Proceedings, Grazing Livestock Nutrition Conference, pp 101-113. Jackson. Wyoming. McCoy, K.E.M. and P.H. Weswig. 1969. Some selenium responses in the rat not related to vitamin E. J. Nutr, 98: 383-389. McDowell, L.R. 1985. Nutrition of Grazing Ruminants in Warm Climates. Academic Press. Orlando. McDowell, L.R., B. Bauer, E. Galdo, M. Roger, J.K. Loosli and J.H. Conrad. 1982. Mineral supplementation of beef cattle in the Bolivian tropics. J. Anim. Sci. 55:964-970. McDowell, L.R. and J.H. Conrad. 1977. Trace mineral nutrition in Latin America. World Anim. Rev. 24:24-33.

PAGE 197

186 McDowell, L.R. , J.H. Conrad and G.L. Ellis. 1984. Mineral deficiencies and imbalances and their diagnosis. Symposium on Herbivore Nutrition in Subtropics and Tropics-Problems and Prospects. Pretoria, South Africa. McDowell, L.R. , J.H. Conrad, G.L. Ellis and J.K. Loosli. 1983. Minerals for grazing ruminants in tropical regions. Extension bulletin, Department of Animal Science, University of Florida, Gainesville. McDowell, L.R.,. J.H. Conrad and J.K. Loosli. 1986. Mineral imbalances and their diagnosis in ruminants. In: International Symposium on the Use of Nuclear Techniques in Studies of Animal Production and Health in Different Environments. Vienna, Austria.. McDowell, L.R., J.H. Conrad, J.E. Thomas, L.E. Harris and K.R. Kick. 1978. Nutritional composition of Latin American forages. Trop. Anim. Prod. 2:273-279. McDowell, L.R., Y. Salih, J.H. Hentges and C.J. Wilcox. 1989. Selenium supplementation and concentration of selenium in cattle tissues and fluids. In: Florida Beef Cattle Report. Gainesville, pp 61-64. Metson, A.J. 1974. Magnesium in New Zealand Soils. I. Some factors governing the availability of soil magnesium: a review. N.Z.J. Exp. Agric. 2:277-319. Metson, A.J., w.M.H. Saunders, T.W. Collie and V.W. Graham. 1966. Chemical composition of pastures in realation to grass tetany in breeding beef cows. N. Z. J. Agric Res. 9:410-436. Michael, D.T. 1962. Manurial treatment in relation to calcium and magnesium levels in sheep. Vet. Record. 74:103-107. Miles, W.H. and L.R. McDowell. 1983. Mineral deficiencies in the llanos rangelands of Colombia. World Anim. Rev. 46:2-10. Millar, K.R. 1983. Selenium. In: N.D. Grace (Ed.) pp 38-43. The mineral requirements of grazing ruminants. Occasional publication No 9. New Zealand Soc. of An. Prod. Palmerston North. New Zealand. Miller, W.J. , J.w. Lassiter and J.B. Jones. 1972. Problems in the use of mineral values for feed formulation. In: Proceedings Georgia Nutrition Conference for Feed Industry, pp 94-106. University of Georgia Press. Atlanta. Miltmore, J.E. and J.L. Mason. 1971. Copper to molybdenum ratio and molybdenum and copper concentrations in ruminant feeds. Can. J. Anim. Sci. 51:193-200.

PAGE 198

187 Minson, D.L. and R. Milford. 1967. The voluntary intake and digestibility of diets containing different proportions of legume and mature Pangola grass ( Diqitaria decumbens ) . Aust. J. Exp. Agr. Anim. Husb. 7:546-551. Moore, J.E. and G.O. Mott. 1974. Recovery of residual organic matter from in vitro digestion of forages. J. Dairv Sci 57:1258-1259. * Montalvo, M.I., J. v. Veiga, L.R. McDowell, w.R. Ocumpaugh and G.O. Mott. 1987. Mineral content of Dwarf Pennisetum purpureum under grazing conditions. Nutrition Reports International. 35:157-169. Moseley, G. and D. J.H. Jones. 1974., The effect of sodium chloride supplementation on the mineral nutrition of sheen Proc. Nutr. Soc. 33:87 A. Mtimuni, J.p. 1982. Identification of mineral deficiencies in soil, plant and animal tissues as constraints to cattle production in Malawi. Ph.D. dissertation. University of Florida, Gainesville. National Research Council (NRC) . 1980. Mineral Tolerance of Domestic Animals. Subcommitee on Mineral Toxicity in Animals. National Academy of Sciences. Washington D.C. National Research Council (NRC). 1985. Nutrient Requirements of Domestic Animals. Nutrient Requirements of Sheep. Sixth Revised Edition. National Research Council, Washington D. C. Perkin-Elmer. 1980. Perkin-Elmer Model 5000 Atomic Absorption Spectrophotometer Operator's Manual. Perkin-Elmer Corporation. Analytical Instruments, Norwalk, Connecticut. Perkin-Elmer. 1984. Zeeman/3 03 Atomic Absorption Spectrophotometer Operator's Manual. Perkin-Elmer Corporation. Analytical Instruments, Norwalk, Connecticut. Peterson, R.G., T.E. Nash and J. A. Shelford. 1982. Heritabilities and genetic correlations for serum and production traits of lactatmg Holsteins. J. Dairy Sci. 65:1556-1561. Pfander, W.H. 1971. Animal nutrition in the tropics: Problems and solutions. J. Anim. Sci. 33:843-849. Pierson, RE. 1966. Zinc deficiency in young lambs. Amer.Vet. Med. Assoc. 149:1279-1284. Pitman, W.D. and E.C. Holt. 1983. Herbage production and quality of grasses with livestock and wild life value of Texas. J. Range Management. 36:52-54.

PAGE 199

188 Poole, D.B.R. 1982. Bovine copper deficiency in Ireland-the clinical disease. Irish Vet. J. 36:169-171. Proyecto Ovino Colombo Britanico. 1979. Informe Anual 1978-1979. ICA, Caja Agraria, Planeacion Nacional, Reino Unido. Boqota! Colombia. Ravel, R. 1989. Clinical Laboratory Medicine: Clinical Application of Laboratory Data. 5 th Ed. Year Book Medical Publishers, Inc. Chicago. Reffet, J.K. and J. A. Boling. 1985. Nutrient utilization in lambs fed diets high in sodium or potassium. J. Anim. Sci 61:1004-1009. Reid, R.L. and D.J. Horvath. 1980. Soil chemistry and mineral problems in farm livestock. A review. Anim. Feed Sci Technol. 5:95-167. Rhue, R.D. and G. Kidder. 1983. Analytical procedures used by the IFAS extension soil testing laboratory and the interpretation of results. Gainesville: Soil Sci. Dent Univ. of Florida. ' Robayo, S.E, J. Cuatrecasas, A. Torres and D. Witlin. 1988 Paramos. Editorial Villegas. Bogota. Rosa, I. V., P.R. Henry and C.B. Ammerman. 1982. Interrelationship of dietary phosphorus, aluminum and iron on performance and tissue mineral composition in lambs. J Anim. Sci. 55:1231-1240. Rosero, O.R., r.e. Tucker, G.E. Mitchell, Jr. and G.T Schelling. 1980. Mineral utilization in sheep fed spring forages of different species, maturity and nitrogen fertility. J. Anim. Sci. 50:128-136. Rowlands, G. J 1980. A review of variations in concentrations of metabolites in the blood of beef and dairy cattle associated with physiology, nutrition and disease. Wld Rev. Nutr. Diet. 35:172-235. Rowlands, G. J. , R. Manston, R.M. Pockock and S.M. Dew 1975 Relationships between stage of lactation and pregnancy ' and blood composition in a herd of dairy cows and the influences or seasonal changes in management on these relationships J Dairy Res. 42:349-362. *»••* Salamanca, S. 1986. La vegetacion del paramo, unica en el mundo. In: Colombia, sus gentes y regiones. Instituto Geografico Agustin Codazzi. Bogota.

PAGE 200

189 Sanchez, P. A. 1976. Properties and management of soils in the tropics. John Wiley and Sons. New York. SAS Institute Inc. 1985. SAS User's Guide: Statistics. Fifth ed. Author, Cary. North Carolina. Schalm, O.W. and C.J. Nemi. 1986. Veterinary Hematology. 4 th ed. Lea and Febiger. Philadelphia. Siddons, R.C. and C.F. Mills. 1981. Glutathione peroxidase ^ activity and erythrocyte stability in calves differing in selenium and vitamin E status. Br. J. Nutr. 46:345-355. Sillanpaa, M. 1982. Micronutrients and the nutrient status of soils: a global study. FAO Soils Bulletin 48. Food and Agriculture Organization of the United Nations. Rome. Spears, J.w. 1989. zinc methionine for ruminants: Relative bioavailability of zinc in lambs and effects of growth and performance of growing heifers. J. Anim. Sci. 67:835-843. Steevens, B. J. , L.J. Bush, J.D. Stout and E.I. Williams. 1971. Effects of varying amounts of calcium and phosphorus in rations for dairy cows. J. Dairy Sci. 54:655-661. Suttle, N.F. 1986. Copper deficiency in ruminants; recent developments. Veterinary Record. 119:519-522. Swift, G. 1972. A review of factors affecting the mineral content of herbage in relation to animal requirements with particular reference to the North of Scotland. Technical Report No l. North of Scotland College of Agriculture. Scotland. Tejada, R. , L.R. McDowell, F.G. Martin and J.H. Conrad. 1987. Evaluation of the macromineral status of cattle in specific regions in Guatemala. Nutrition Reports International. 35:989-998. Tomas, F.M. and B.J. Potter. 1976. The effect and site of action of potassium upon magnesium absorption in sheep. Australian J. Agr. Res. 27:873-878. Towers, N.R. and R.G. Clark. 1983. Factors in diagnosing mineral deficiencies. In: The mineral requirements of grazing ruminants. Occasional Publication No 9. New Zealand Society of Animal Production. Ullrey, D.E. 1987. Biochemical and physiological indicators of selenium status in animals. J. Anim. Sci. 65:1712-1726. Underwood, E.J. 1981. The Mineral Nutrition of Livestock (2nd ed.). Commonwealth Agricultural Bureau, Farnham Royal, England.

PAGE 201

190 Valdes, J.L. , L.R. McDowell and M. Koger. 1988. Mineral status and supplementation of grazing beef cattle under tropical conditions in Guatemala. II. Microelements and animal performance. J. Prod. Agric. 1:351-355. Vargas, R. , L.R. McDowell, J.H. Conrad, F.G. Martin, C. Buergelt and G.L. Ellis. 1984. The mineral status of cattle in Colombia as related to a wasting disease ("secadera") . Trop. Anim. Prod. 9:103-113. Velez, J. and W.G. Blue. 1971. Effect of lime and extractable iron and aluminum, and phosphorus absorption in a tropical and a temperate soil. Proc. Soil Crop Sci. Soc. Florida 31:169-174. Viets, F.G. and W.L. Lindsay. 1973^ Testing soils for zinc, copper, manganese and iron. In: L.M. Walsh and J.D. Beaton (Ed.). Soil testing and plant analysis. Soil Science Society of America, Inc. pp 153-172. Madison. Wisconsin. Volkweiss, S.J. 1978. Soil properties that influence mineral deficiencies or toxicities in plants and animals. In: J.H. Conrad and L.R. McDowell (Ed.). Latin American Symposium on Mineral Nutrition Research with Grazing Ruminants, pp 17-22. University of Florida, Gainesville. Ward, G.M. 1978. Molybdenum toxicity and hypocuprosis in ruminants: A review. J. Anim. Sci. 46:1078-1085. Watt, A.T. 1978. The biology of Holcus lanatus L. (Yorkshire fog) and its significance in grassland. Herbage Abstracts. 48:195-204. Whanger, P.D., P.H. Weswig, J. A. Schmitz and J.E. Oldfield. 1978. Effects of various methods of selenium administration on white muscle disease, glutathione peroxidase and plasma enzyme activities in sheep. J. Anim. Sci. 47:1157-1166. Whetter, P. A. and D.E. Ullrey. 1978. Improved fluorometric methods of determining selenium. J. Assoc. Anal. Chem. 61:927-930. White, C.L. , T.K. Caldwalader, W.G. Hoekstra and A.L. Pope. 1989. Effects of copper and molybdenum supplements on the copper and selenium status of pregnant ewes and lambs. J. Anim. Sci. 67:803-809. Wilkinson, H.F. 1972. Movement of micronutrients to plant roots. In: J.J. Motvedt, P.M. Giordiano and W.L. Lindsay (Ed.). Micronutrients in Agriculture, pp 139-169. Soil Sci Soc. Am. Madison. Wisconsin.

PAGE 202

191 Williams, R. 1963. Minor elements and their effects on the growth and chemical composition of herbage plants. Publication No 1/1959. Commonwealth Agricultural Bureau, Bucks, England.

PAGE 203

BIOGRAPHICAL SKETCH Rodrigo Pastrana was born in Neiva (Huila) , Colombia, on February 27, 1945, the son of the late Luis Carlos Pastrana and Solita Bonilla dePastrana. He attended the National University of Colombia and graduated with a doctoral degree in veterinary medicine and zootechnics in 1967. Since then, he has been working in the sheep program of the Instituto Colombiano Agropecuario (ICA) in Bogota. In 1971, he received a master's degree in animal science from the University of Wyoming. In 1978 he was awarded a diploma degree in tropical animal health and production at the University of Edinburgh, Scotland. With the support of a scholarship from Instituto Colombiano Agropecuario, he enrolled in August 1986 in the Graduate School at the University of Florida and is a Ph.D. candidate. He is married to Diana Herrera de Pastrana and has five children, Diana Constanza, Gloria Consuelo, Maria Paola, Rodrigo Jose and Ian Gabriel. 192

PAGE 204

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. 4 /?. a '/ V4A .Lee R. McDowell, Chairman 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 the degree of Doctor of Philosophy. Clarence B. Ammerman Professor of Animal Science

PAGE 205

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. tkjL=> 8. ^sfe Dfam ouglas B. Bates 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. / -U-?\^T\ e Lynn E. Sollenberger /' Assistant Professor of Agronomy 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. December 1989 Lax 3 "%L Dean, (ipollege of Agriculture Dean, Graduate School

PAGE 206

UNIVERSITY OF FLORIDA lllllllllffllllllll 3 1262 08554 8583