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Effect of Long Term Use of Bovine Somatotropic Hormone on Milk production, Reproduction, Nutrient Utilization and Blood metabolites in Nili-Ravi Buffaloes BY IFTIKHAR AHMAD 2004-VA-01 A thesis submitted in partial fulfillment of requirements for degree of Doctor of Philosophy in Animal Nutrition UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, LAHORE 2009

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Effect of Long Term Use of Bovine Somatotropic Hormone on Milk production, Reproduction, Nutrient Utilization and Blood

metabolites in Nili-Ravi Buffaloes

BY

IFTIKHAR AHMAD

2004-VA-01

A thesis submitted in partial fulfillment of requirements for degree of Doctor of Philosophy in Animal Nutrition

UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, LAHORE

2009

II

III

IV

Effect of Long Term Use of Bovine Somatotropic Hormone on milk Production, Reproduction, Nutrient Utilization and Blood

metabolites in Nili-Ravi Buffaloes

BY

IFTIKHAR AHMAD 2004-VA-01

A thesis submitted in partial fulfillment of requirements for degree of

Doctor of Philosophy in Animal Nutrition

UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, LAHORE

2009

V

To

The Controller of Examinations

University of Veterinary and Animal Sciences,

Lahore.

We the members of supervisory committee certify that the contents and form of the

thesis submitted by Mr. Iftikhar Ahmad has been found satisfactory and recommend that it

be processed for evaluation by external examiner (s) for the award of degree.

CHAIRMAN -----------------------

MEMBER---------------------------

MEMBER---------------------------

VI

ACKNOWLEDGEMENT

O ver above every th ing I offer m y hum ble and sincerest gratitude to A L M G H T Y A L M G H T Y A L M G H T Y A L M G H T Y

A L L A H ,A L L A H ,A L L A H ,A L L A H , the m ost M ercifu l and Com passionate, the m ost G racious and B eneficent, W ho

bestow ed on the deep oceans of know ledge already existing. I offer m y hum blest thanks; form the

deepest of m y heart to the H oly Prophet, M U H A M M A DH oly Prophet, M U H A M M A DH oly Prophet, M U H A M M A DH oly Prophet, M U H A M M A D (peace be upon h im ) w ho is, forever a

torch of gu idance and know ledge for hum anity as a w hole.

This m anuscript w ould not have been possible w ithout the enthusiasm , know ledge,

gu idance, tenacity, and, perhaps m ost im portantly faith that I received from m y supervisor,

P rofessor D r. M akhdoom A bdul Jbbar, Chairm an, F ood and N utrition D epartm ent, P rofessor D r. M akhdoom A bdul Jbbar, Chairm an, F ood and N utrition D epartm ent, P rofessor D r. M akhdoom A bdul Jbbar, Chairm an, F ood and N utrition D epartm ent, P rofessor D r. M akhdoom A bdul Jbbar, Chairm an, F ood and N utrition D epartm ent, F rom the

very first interv iew to the last queries on research accom plishm ents and career p lans, he w as

alw ays eager to entertain m y ideas in hope that I fulfilled m y dream (s) and becom e successfu l. I

w as not sure I could handle an undertaking of such a m agnitude, but w as able to thanks to h is

consistent effort and true desire to keep m e on track .

I am greatly indebted to, P rofessor D r. T alat N aseer P asha D ean F aculty of A nim al P rofessor D r. T alat N aseer P asha D ean F aculty of A nim al P rofessor D r. T alat N aseer P asha D ean F aculty of A nim al P rofessor D r. T alat N aseer P asha D ean F aculty of A nim al

production and Technologyproduction and Technologyproduction and Technologyproduction and Technology and P rofessor D r. M uham m ad A bdullah D epartm ent of L ivestock P rofessor D r. M uham m ad A bdullah D epartm ent of L ivestock P rofessor D r. M uham m ad A bdullah D epartm ent of L ivestock P rofessor D r. M uham m ad A bdullah D epartm ent of L ivestock

P roductionP roductionP roductionP roduction for their great help painstaking contribution during the course of research and

finalization of th is w rite up .

I ow e a debt of gratitude to D r. R ashid A hm ad, D irectorD r. R ashid A hm ad, D irectorD r. R ashid A hm ad, D irectorD r. R ashid A hm ad, D irector L ivestock Production R esearch

Institute, B ahadurnagar, O kara and R aja M uham m ad L atifeR aja M uham m ad L atifeR aja M uham m ad L atifeR aja M uham m ad L atife, Sen ior R esearch O fficer, A nim al

N utrition Center, R akh D era Chahal, L ahore for their generous co- operation , help and provid ing

an environm ent conducive for a creative activ ity like research .

I am also thankful to D r. K halid JavidD r. K halid JavidD r. K halid JavidD r. K halid Javid , A ssociate Professor D epartm ent of A nim al

B reeding and G enetics for h is help during the w rite up phase. I feel p leasure to p lace on record m y

profound obligation to D r. Sualeha R affitD r. Sualeha R affitD r. Sualeha R affitD r. Sualeha R affit, A ssistant Professor, D epartm ent of Pharm acology,

U niversity of V eterinary and A nim al Sciences, L ahore for her active cooperation during m y Ph.D .

studies.

VII

L ast but not the least, I am greatly indebted to m y brothers, for their love, im m ense

orison , w hich heartened m e to achieve success in every sphere of life and w hose hands alw ays

rem ained raised in prayer for m e.

(IF T IK H A R A H M A D )(IF T IK H A R A H M A D )(IF T IK H A R A H M A D )(IF T IK H A R A H M A D )

VIII

CONTENTS

CHAPTERS T I T L E Page

List of tables IX

List of figures XI

1 Introduction 1

2 Review of literature 6

3

Effect of long-term use of bovine somatotropic hormone on

milk production, Reproduction and physiological parameters in

Nili Ravi Buffaloes

Abstract

Introduction

Materials and Methods

Results and discussion

31

31

32

33

40

4

Production Performance of Lactating Nili-Ravi Buffaloes

under the influence of Somatotropic Hormone on varying

levels of dietary Energy

Abstract

Introduction

Materials and Methods

Results and discussion

64

64

65

66

70

5

Publications

I Effect of bovine somatotropic hormone on the

productive Performance of Nili-Ravi buffaloes

II Performance of lactating buffaloes administered bovine

somatotropin hormone

76

81

6 Summary 84

Literature cited 88

IX

LIST OF TABLES Table No

T I T L E

Page

1 Average chemical composition of fodders (self analyzed in

nutrition labs. at University of Veterinary and Animal Sciences,

Lahore) used to fulfill the maintenance requirements of

experimental animals during study period.

36

2 Composition of Concentrate Ration used to fulfill the production

requirements of treated (+bST) and control (without bST) during

study.

37

3 Effect of bovine somatotropic hormone administration on Milk

yield of Nili- Ravi buffaloes (means ± SE)

45

4 Effect of recombinant bovine somatotropic hormone on

productive traits of Nili- Ravi Buffaloes (means ± SE).

47

5 Effect of bovine somatotropic hormone administration on milk

components of lactating Nili –Ravi buffaloes over the 3 year

study period

50

6 Effect of bovine somatotropic hormone administration on

reproductive parameters of lactating Nili –Ravi buffaloes during

study period.

54

7 Effect of bovine somatotropic hormone administration on health of lactating Nili- Ravi buffaloes

57

X

8 Effect of bovine somatotropic hormone administration on

hematology of lactating Nili –Ravi buffaloes (Means ±SE)

58

9 Administration effect of bovine somatotropic hormone on blood

metabolites in lactating Nili –Ravi buffaloes (Means ±SE).

60

10 Effect of bovine somatotropic hormone administration on mineral

metabolites of lactating Nili –Ravi buffaloes (Means ±SE)

61

11 Economics of use of bovine somatropic hormone in lactating Nili-

Ravi buffaloes.

63

12

Ingredients and Chemical Composition of Total Mixed Rations

(TMR) offered with administration of bST in Nili Ravi buffaloes.

67

13

Effect of bovine somatotropic hormone administration on

productive performance with in lactating Nili Ravi buffaloes fed

varying level of dietary energy (means± SE).

72

14 Effect bovine somatotropic hormone adminstration on daily nutrients intake of in Nili Ravi buffaloes fed varying level of dietary energy (means± SES).

75

15 Mean (± S.E.) values of different haematological and biochemical

parameters.

79

16 Intake and digestibility of nutrients in buffaloes administered

Bovine somatotropin hormone.

81

17 Performance of buffaloes on bST administration.

82

XI

LIST OF FIGURES

S.No T I T L E Page

1 Monsanto mechanism of recombinant bovine somatotropic

hormone

8

1 Trend of milk production in control and bST- treated groups

during the first year of the study

43

2 Milk production pattern in control and bST- treated groups

during the 2nd year of the study.

44

3 Trend of milk production in the control and bST-treated

groups during the 3rd year of the study.

44

4 Comparison of milk production in control and bST-treated

groups over the 3 year study period.

45

5 Comparison of lactation length (LP), dry period (DP) and

calving interval (CI) in bST-treated and control groups during

the study period

48

6 Comparison of yearly and overall milk fat % of bST-treated

and control groups over the study period

51

7 Comparison of yearly and overall solid not fat % of bST-treated

and control groups during the study period.

51

8 Comparison of year wise and overall total solids % of bST-

treated and control groups during the study period.

52

9 Comparison of reproduction parameters (post partum estrus

[PPE], service period [SP]) of bST-treated and control groups

during the study period.

55

10 Trend of milk production in animals given low, medium and

high-energy diets.

71

11 Average feed efficiency of bST-treated Nili-Ravi buffaloes fed

low, medium and high energy density rations

74

1

Chapter-1

INTRODUCTION

Adaptation of recent developments in various scientif ic and

technological f ields is important to sustain the growing world

population, which is predictable to reach 8000 million by 2020, of

which about 6700 mill ions will be in developing countries (Mehra,

2001). It has been estimated that the supply of food required to

adequately meet human nutritional needs over the next 40 years is

quantitatively equal to the amount of food previously produced

throughout the history of mankind (NRC, 1994). With f inite resources

and an increasingly vulnerable environment, i t is highly important that

eff icient growth of agriculture and livestock products should be a

dominant factor. To meet future needs and to sustain agricultural

production, agricultural research and its applications wil l have to use

all available technologies and especially the rapidly developing modern

biotechnologies that increase productive eff iciency of food production.

With respect to animal production, synthetic bovine somatotropic

hormone (bST) is one of biotechnological product that increases the

food production per unit of feed intake (Bauman, 1992).

Bovine somatotropic hormone is produced by the somatotrophs of

the anterior pituitary gland and consists of a chain of 190 to 191 amino

acids (Hedge et al. , 1987). The secretion of bovine somatotropic

hormone is regulated by two well-known hypothalamic peptides that

proceed to motivate (growth hormone-releasing factor) or diminish

(somatostatin) the release of ST from the pituitary gland (Tuggle and

Trenkle, 1996). The amino acid sequence of bST is substantially

2

different from human ST (35% of the amino acids in hST vary from

bST). For this difference, bST has no effect on human development, in

harmony with their binding aff inity to the hST receptor being several

orders of magnitude lower than that of hST (Lesniak et al., 1977).

Bovine somatotropic hormone is released from the pituitary as

four variants. These variants have either a leucine or valine substitution

at posit ion 127 and an alanine or a phenylalanine at the NH2 terminus

(Wood et al., 1989). The variation in the NH2 terminus is due to

differences in cleavage of the signal peptide. The frequency of these

gene alleles differs between dairy breeds (Lucy et al., 1993).

Furthermore, there are some signs that these variants may differ in their

effectiveness. Although studies have been limited, results indicate that

treatment with the valine-127 variant elicited a greater increase in

circulating ST and milk yield than the leucine-127 variant (Eppard et

al. , 1992). The commercial bST formulation (Monsanto, St. Louis, MO)

approved for apply in dairy cows is leucine variant-127, it has an extra

methionine at the NH2 terminus.

Somatotropic hormone was first characterized in the 1920s when

Evans and Simpson demonstrated growth-promoting effects in rats

treated with a crude extract from bovine pituitaries. The discovery that

the pituitary contained a factor that stimulated growth led to this factor

being referred to as "growth hormone." It became apparent shortly

thereafter, however, that growth hormone did much more than

stimulating growth, since administration of pituitary extracts also

enhanced milk yield in lactating goats (Asdell, 1932). Major research

development in ST study was accomplished in 1937, when Russian

scientists treated dairy cows with BST and demonstrated that milk yield

was increased with the administration of pituitary extract (Asimov and

3

Krouze, 1937). Studies conducted in the 1940s showed that BST

augmented the milk yield of dairy cows; however, the amount of BST

that could be extracted from the pituitary glands of slaughtered animals

was inadequate to impact commercial production (Young, 1947). In

1982, bST was produced by DNA biotechnology that permitted

scientists around the world to observe various aspects of its biology

(Bauman, 1992).

The dependable genes for production of BST in catt le were

identif ied in bovine t issue cells, which cause the pituitary cells to

produce the biological product of BST. These genes were isolated and

inserted into specif ic bacteria as a part of plasmid, as these altered

bacteria replicate, the new genes are also replicated and conceded along

to all new bacteria. The presence of these genes causes the bacterial

cell to become a lit t le manufacturing plant, which produces bST in

large quantities. Eventually the bacterial cells are kil led and removed,

leaving the purif ied bST (Bauman et al., 1985; Roush, 1991).

Somatotropic hormone orchestrates many diverse physiological

processes, so that for milk synthesis (during lactation) additional

nutrients can be uti l ized. Somatotropic hormone is a homeorhetic

control that affects numerous target tissues in ways that are highly

coordinated to affect marked changes in nutrient partitioning among

these tissues. The biological effects of somatotropic hormone can be

broadly classif ied as either somatogenic or metabolic. The somatogenic

effects are those in which ST stimulates cell proliferation. These effects

are mediated by insulin-l ike growth factor (IGF-I) (Rechler and

Nissley, 1990).

Milk yield response to bST has been observed for all dairy breeds

and in animals of different parity and genetic potential (Burton et al. ,

4

1994). In general, milk yield response in early lactation is very small,

and thus bST use is usually over the last 80% of the lactation cycle.

Typical milk yield responses are increases of 10-15% although even

greater increases occur when the management and care of the animals

are excellent (Chil l iard, 1989; NRC, 1994).

Bovine somatotropic hormones improve the productive eff iciency

because administration of bST to dairy animals increases milk yield per

unit of feed. In dairy cattle, use of bST has resulted in improvements

of 12-25% in milk yield without any substantial alteration in the

composition of milk (Ludri et al., 1989). BST hormone reduces the

proportion of maintenance requirement out of total nutrient intake

(Bauman et al., 1989). The higher productive response is attributed to

changes in coordinated metabolism of various body organs and tissues

that support increased synthesis of lactose, fat and protein in mammary

glands, in meticulous the glucose and fatty acid metabolism that

account for increase in lactose and milk fat using bST treatment

(Bauman et al., 1988). An addit ional beneficial effect is improvement

in pregnancy rates when bST is injected during the period prior to

estrus and subsequent artif icial insemination (Santos et al. , 2004).

Most of available studies on bST hormone are related to cattle,

however, buffalo is the main dairy animal in Asia and 97% of the world

population is found in Asian subcontinent (FAO, 2004). Of the total

worldwide milk production 12.84% comes from buffalo, with 97.11% of

buffalo milk produced in the developing countries of Asia. Over the

last decade the annual growth in buffalo milk production was 6.19%

higher than that of worldwide total milk production (FAO, 2003),

highlighting the importance of buffalo as a dairy animal. In spite of

large numbers and good adaptabil ity, per animal productivity is very

5

low as compared to Western dairy animals. The normal lactation yield

in the best-known dairy breeds eg. Nil i-Ravi and Murrah is around

2000 kg (Agarwal and Tomar, 1998).

There are multiple approaches to improve productivity of the

dairy buffalo including improvement in genetics, nutrition and

management. Use of bST is one way for increasing milk production in

dairy animals. Usmani et al. (1991) have suggested that research should

be planned to determine the effect of bST in buffaloes before its

commercialization in Pakistan. However, since then some researchers

have undertaken short-term studies involving buffaloes but

comprehensive information is stil l lacking. The present project was

planned to ascertain the consequences of extensive use of bST on

productive and reproductive parameters in lactating Nili-Ravi buffaloes

with the following objectives

a) To study the long term effect of bST hormone on production,

reproduction and physiological parameters in Nil i-Ravi buffaloes.

b) To monitor the quality of milk under the influence of bST through

chemical composition.

c) To study the effect of bST on nutrient uti l ization in Nili- Ravi

buffaloes.

d) To determine the optimum dietary energy levels for eff icient milk

production using bST in Nil i-Ravi buffaloes.

6

Chapter-2

REVIEW OF LITERATURE

2.1 Bovine somatotropic hormone

The effects of somatotropic hormone on the performance of

animals have been the subject to interest for over 50 years. Early

research established that injection of crude pituitary extracts increased

growth rate in rats. This extract was called somatotropin, a Greek word

meaning “tissue growth”. Somatotropin is a hormone that is produced

by one organ and transported to another to cause a biological effect. In

contrast to steroid hormone it is composed of 20 different amino acids,

which are combined in specif ic sequence to form different body

proteins, however, every species has its own amino acid sequence

(Kostyo and Reagan, 1976 ). Being a proteinous hormone in nature it

must be injected for biological activity and if it is given orally the

hormone is broken down in to amino acids during the digestive process

like other dietary proteins (Juskevich and Guyer, 1990). In order to

produce biological effect, protein hormone must f irst bind to specif ic

receptors located at the cell surface. The amino acid sequence develops

its three dimensional shape, which determines whether the protein wil l

be able to bind to t issue and elicit a biological response. On the

identif ication of the amino acid sequence it becomes clear that the three

dimensional unique shape of bovine somatotropic hormone (BST)

differs from that of human somatotopin (hST). This means that it is not

possible to bind BST receptors to human tissues (Wallis, 1975).

However, BST is biologically active in sheep because the amino acid

composition of ovine somatropin and BST differ only by one amino

acid. BST, which is naturally produced in cows, has homeorhetic

7

control that coordinates the metabolism of many tissues in dairy cattle

(Bauman et al., 1988).

2.2 Recombinant bovine somatotropic hormone (bST)

Prior to 1980s restricted progress was made in BST research

due to limited availabil ity and its high cost of production because it

was extracted in only small quantities with varying purity from each

pituitary gland of slaughtered animals. Thus only a limited number

of dairy animals were able to be treated for only a l imited period of

time (Peel and Bauman, 1987). The advent of recombinant DNA

technology allowed the production of large quantit ies of synthetic

bST. Since then a lot of research has been conducted on the effect of

bST on different production parameters (Peel et al. , 1985; McGuffy

et al. , 1990). Bauman et al. (1982) were the foremost researchers

who util ized the recombinant methionyl bovine somatotropic

hormone and showed that i t was effective as a pituitary derived

growth hormone in enhancing the milk production. Recombinant

bovine somatotropic hormone (bST) is a genetically engineered

synthetic analogue of the natural hormone. Scientists identif ied the

gene responsible for the production of natural BST and by using

standard genetic–engineering techniques (Fig 1), duplicated the gene

and spliced it into the DNA of Escherichia coli (E. coli) bacterium.

These bacteria can be grown in fermentation tanks and bST is

produced with the organism. The E. coli are killed and ruptured and

large amounts of bST can then be inexpensively extracted for

injection into the cow (Bauman et al., 1985; Roush, 1991). The

injection of bST produces a biological reaction that is essentially the

same as that which occurs in dairy cows that naturally produce

higher levels of BST (Bauman et al. , 1985).

8

Figure I. Monsanto mechanism for the production of recombinant

bovine somatotropic hormone (www.diseaseproof.com)

9

2.3 Mechanism of action of bST

Somatotropic hormone, through homeorhetic control, shifts the

partitioning of nutrients in a lactating cow so that more nutrients are

used for milk synthesis (Bauman, 1992). This involves coordinating the

metabolism of diverse body organs and tissues and includes the

metabolism of al l nutrients (carbohydrates, lipids, proteins and

minerals). Thus, treatment with bST not only increases the rate of milk

synthesis within the mammary gland but also orchestrates other body

processes in a manner to provide the necessary nutrients to prop up this

enhanced rate of milk synthesis. Baldwin and Knapp (1993)

summarized a number of the major coordinated changes that occur with

bST treatment of a dairy cow. These adaptations are of critical

importance during the init ial period of bST treatment when milk

production is augmented but feed intake is not affected. These

adaptations can be broadly divided into two types (i) direct effects on

some tissues; and (i i) indirect effects that are thought to be mediated by

the insulin-l ike growth factor (IGF) system (Bauman and Vernon,

1993).

Direct actions of ST appear to be primarily concerned with the

coordination of metabolic processes (Bauman, 1992; Baldwin and

Knapp, 1993). Adipose tissue provides an example to demonstrate the

mechanisms of action. Adipose tissue has two main functions,

lipogenesis and lipolysis. Somatotropic hormone treatment has no acute

effects on either of these functions, but it does alter l ipid metabolism

on a chronic basis. In particular, ST treatment changes adipose tissue

retort to homeostatic signals affecting lipid synthesis and l ipid

mobil ization. Studies have shown that somatotropic hormone reduces

10

the abil ity of insulin to stimulate lipid synthesis and enhances the

abili ty of catecholamine to stimulate l ipolysis. Thus, if the cow is in a

posit ive energy balance, bST treatment reduces the nutrients used for

body fat and these nutrients are redirected in support of the increased

milk synthesis (Bauman and Vernon, 1993). On the other hand, if the

cow is near zero or in negative energy balance when bST treatment is

initiated, then body fat reserves are mobil ized to support the need of

nutrients for the additional milk production. With prolonged

Somatotropic hormone treatment, voluntary food intake increases and

animals return to a positive energy balance allowing the replenishment

of body reserves over the lactation cycle (Bauman et al ., 1988)

The indirect effects of ST appear to be primarily associated with

the mammary gland through the actions of IGF system (Bauman and

Vernon, 1993). These effects involve an increase in the rate of milk

synthesis per cell and an improved maintenance of mammary cells. The

IGF system is complex involving IGF-1 and IGF-1I as well as two

specif ic IGF receptor types (Celemmons and Underwood, 1991). In

addit ion, the preponderance of IGFs in body f luids is bound to soluble,

high aff inity binding proteins. There are six specif ic IGF-binding

proteins (IGFBP) and their postulated roles include serving as

circulatory transport vehicles, retarding IGF degradation, facilitating

transvascular movement, providing an extra vascular pool, and

modulating directly the actions of IGFs at specif ic marked cells.

Administration of bST to well-managed lactating cows causes an

increase in circulating concentrations of IGF-1 and IGFBP-3 and a

decrease in IGFBP-2 (Bauman and Vernon, 1993).

11

2.4 Milk production and bST

Administration of bST has been reported to enhance the lactation

performance in all dairy breeds but milk yield response to bST is

dependent on the dosage level, parity and genetic potential of the

animal (Burton et al. , 1994). It must be given each day to maintain an

increased milk response because it is not stored in the body and is

rapidly cleared from the blood stream. To amplify milk yield bST must

be administered daily or in prolonged release formulation. Ludri and

Singh, (1989) reported increased milk yield over controls by 19.4 and

32.39% for 25 and 50 mg bST/d, respectively in Murrah buffaloes. In

dairy cows increases in milk production with similar protocols ranged

from 15 to 25%. Similarly, Chil l irad (1989) summarized the conclusion

of different studies and reported that daily milk yield increased 2.8,

3.9, 5.2 and 5.6 kg/d against 5, 10, 20 and 31 mg bST/d, respectively.

In the same way super-physiological doses of Sometribove have been

shown to increase milk production. Eppard et al. (1991) evaluated the

effect of bST in a sustained-release formulation administered to

lactating cows at concentrations up to 3.0 g every 14 d over two

lactations. During the f irst and 2nd year of treatment, administration of

bST increased mean 3.5% fat-correct milk (FCM) by 21.1 and 24.8 kg/d

respectively over the control.

Other researchers have also reported increased milk yield in

response to bST administration. Phipps et al. (1997) investigated the

effects of administration of a prolonged-release formulation of bST at

14 d intervals on milk production of indigenous, crossbred, and

purebred cows in Africa. Administration of 334 mg bST increased milk

from 22 to 25% in various breeds of cattle; however, increasing the

12

dose by 500 mg did not further increase milk production. To determine

the breed difference in milk yield response to bST, Jorge et al. (2002)

studied the effect of administering 500 mg bST every 14 d for a period

of seven months on milk production of Murrah buffaloes and found that

milk production was increased in the treated group by 15.72% on total

milk yield, adjusted 4% of fat corrected milk (FCM). Similarly, Mishra

and Shukla (2004) investigated the effect of bST on milk yield with

normal recommended dose of 250mg/14 d in lactating buffaloes and

found a 25% increment in milk yield.

The milk yield response to bST is generally dose dependent and

related to lactation period. Mc Guffy et al. (1991) evaluated lactation

performance of multiparous Holsteins cows supplemented with 0, 320,

640, or 960 mg of Sometribove every 28 d. The treatment period was of

various lengths, depending upon stage of lactation of animals at f irst

administration (of bST). Milk and 3.5% FCM yields were increased by

each dose of Sometribove in all stages.

Whilst administering bST increases milk production in the current

lactation, the effects of long-term administration of bST are less clear.

Hansen et al. (1994) conducted a long-term study to observe the effects

of bST in herds of Holstein cows. bST was injected daily from 28 to 35

d postpartum with 0 (control), 5.15, 10.3, and 16.5 mg bST/d for a two-

year period. During the f irst year FCM production increased l inearly in

response to increased bST administration; however, the magnitude of

this increase decreased in the second year to only 67% of the response

during the f irst year.

Posit ive responses to bST have been reported during short term

studies for other species. Baldi (1999) reported increased milk yield

13

(20-30%) in dairy ewes following treatment with bST. Similarly in

dairy goats, administration of bST increased overall milk yield by 14-

29%. In Ital ian river buffalo bST treatment increased milk yield by

about 17%. In general, studies on dairy ruminants show that treatment

with bST increased milk production in the short term (immediate post

injection period) and that there is also a medium to long term effect on

persistency of lactation (Huber et al. , 1997).

Use of bST during the transit ion period may cause positive and

beneficial effects supporting milk synthesis prior to parturition. Milk

production response to bST occurs because of its known effects on

partitioning of nutrients and because a greater proportion of the

nutrient intake is used for milk synthesis. It increases liver glucose

output, cardiac output, blood f low to the mammary gland and uptake of

nutrients used for milk synthesis by the mammary gland among other

effects. In addit ion, ST decreases the rate of oxidation of amino acid

and glucose and therefore decreases glucose clearance (Gulay and

Hatpolu, 2005). Treatment with bST results in coordinated changes of

various organs and tissues that naturally occur during the transit ion

from a non-lactating to lactating state when circulating concentration of

ST is high. Gulay et al. (2004) investigated the effect of low doses of

bST during the transition period and early lactation period and their

results suggested that administration of low dosage of bST during the

transit ion and early lactation period resulted in increases in milk yield.

In early lactation there is typically an extensive period of

negative energy balance. There are signif icant responses in milk yield

to bST administration, especially when used in combination with

feeding of high energy density rations Polidori et al . (1997) evaluated

14

the effects of bST and calcium salts of long chain fatty acids during

early lactation of Italian river buffaloes and found that bST along with

high energy diets had a beneficial effect on milk yield. Moalllem et al .

(2000) allocated high yielding, individually fed cows to three

treatments or a control group. Calcium soap of fatty acids was fed at

2.2% DM, and bST injected from 10 - 150 days-in-milk (DIM). They

found that treating high yielding cows with bST in early lactation

increased milk production at the expense of an extensive period of

negative energy balance. However, Vicini et al. (1994), in investigating

the effect of supplemental energy and protein on milk production

response to bST in cows found that additional energy and protein did

not alter the milk production response to bST. Similarly, Bremmer et

al. (1996) found that additional ruminally-protected amino acids and fat

in combination with bST resulted in no response in milk production in

cows fed a well balanced diet.

Several researchers have investigated the effects of bST when

used in combination with dietary treatments. Nisa et al. (2006)

investigated the effect of bST when used in combination with varying

levels of enzose on milk yield and its composit ion in lactating Nili-

Ravi buffaloes. They found the interactions for bST x Enzose were

signif icant only for milk yield and milk ash contents (P<0.025), with

bST administration increasing milk production by 30% when used in

combination with 20% enzose. Abo El-Nor et al. (2007) evaluated the

effects of bST and monensin on the milk yield in buffaloes. The control

group of animals was fed a basic ration, the 2nd group was also fed the

basic ration and injected with bST, the 3rd group was fed the basic

ration and supplemented with 4 g monensin/hd/d whilst the 4th group

was fed a supplemented ration as well as being administered bST.

15

Animals treated with bST had signif icantly higher milk yield (p<0.01),

and this was attributed to increased production of propionate in the

rumen. Monensin treatment caused increased production of insulin by

the pancreas, resulting in a decreasing effect on the production of

somatotropin and thus diverting nutrients to body tissues rather than to

milk production. Khattab et al. (2008) evaluated the singular and

combined effect of bST and monensin on the productive performance of

buffaloes. They found that treatment with bST resulted in signif icant

production benefits including higher milk yield and 4% FCM yield and

importantly, without any adverse effects on the health of lactating

buffaloes.

Management and nutrit ion are major factors that affect responses

in lactation yield and its persistency to bST administration. A poorly

managed herd is l ikely to achieve li t tle or no benefit f rom bST use

(Bauman, 1989). Mollet et al. (1986) reported that deficiencies in

environmental, nutrit ional and milking management programs may

attributed to the lack of response to bST. In investigating the

interaction of the effects of photoperiod and bST on milk production

Miller et al. (1999) found that a combination of bST with long daily

photoperiod tended to amplify the increases in milk production. Van

Baale et al. (2005) determined the effects of bST on early lactation

milk yield and subsequent lactation persistency with or without bST

Treatments included a control group milked 3x and three groups milked

6x for either the f irst 7, 14, or 21 DIM. These four groups of cows all

received bST starting at 63 DIM. The fif th treatment group was also

milked 6x for the f irst 21 DIM but these cows received no bST during

the entire lactation. Cows subsequently administered bST (at 63 DIM)

16

produced more milk (38.8 vs. 34.2 ± 0.9 kg/d) than the group injected

in early lactation.

2.5 Milk composition and bST

Like milk yield, bST may also affect milk composit ion, but

mostly the effect is only minor. Bauman et al. (1989) evaluated the

eff icacy of a prolonged-release formulation for sometribove (n-

methionyl bST) in Holstein cows. Milk contents of lactose, fat, ash,

and calcium were not affected in primiparous and multiparous animals

but protein and phosphorous were slightly greater (less than 5%) in

milk from sometribove-treated cows. In contrast, Barbano et al. (1992)

found variable effect of bST on milk composit ion. The

concentrations of milk lactose, fat and ash were similar from cows

receiving bST or excipient; however, percentages of non-protein

nitrogen (NPN) and total protein were greater in milk from bST-treated

cows compared with milk from cows injected with excipient. Lynch et

al. (1992) injected Holstein cows bi-weekly (from 60 ± 3 d postpartum

and continued at 14-d intervals for the ful l lactation) and found that

whilst production of milk, total fat, and all milk fat components

measured were increased, the average fatty acid composit ion of milk fat

remained unchanged. Phospholipids and cholesterol content of milk

changed with stage of lactation but were not influenced by bST

treatment. They concluded that the composit ion of milk is more

influenced by dietary manipulation than by administration of bST.

Stegeman et al. (1992) evaluated the interactions between bST and

dietary fat supplementation (either from sunflower or saff lower seeds)

on the composition and flavor of milk and butter. Supplementation with

unsaturated dietary fat resulted in lower concentrations of short- and

17

medium-chain and higher concentrations of long-chain fatty acids in

milk fat and butter. Sensory evaluations indicated that butters from the

bST with sunflower seed and bST with saff lower seed treatments were

equal or superior in f lavor to the butter from the control cows. Milk

from cows receiving bST or supplemented with unsaturated dietary fat

and receiving bST was no more susceptible to oxidized off-f lavors than

control milk.

Responses in milk composition are often mediated by the energy

status of the cows. Capuco et al. (2001) investigated the relationship

between energy balance and effects of bovine somatotropin (bST) on

thyroid hormone metabolism and cytokine concentrations in serum.

Cows were fed to achieve either positive or negative energy balance

and then administered 40 mg bST/d. Whilst bST decreased energy

balance in both groups of animals, milk production and the fat

percentage of milk increased in response to bST treatment in both

dietary groups. Other researchers have investigated potential

relationships between bST and other hormones in the dairy animal.

Molento et al. (2002) investigated the effects of insulin, bST, and their

interaction on milk protein and selected blood parameters in Holstein

cows. Percent milk protein and urea contents were found to be lower

during bST administration, regardless of insulin infusion.

Micro-constituents of milk are also unchanged under the

influence of bST. For example, milk from bST-treated cows does not

differ in vitamin content or in concentrations of nutritionally important

mineral elements (Bauman, 1992). Long-term studies conducted at four

different locations (Missouri, New York, Maryland, and Utah)

involving a total of 364 cows indicated that the percentage of total ash,

phosphorous and calcium in milk was not altered by bST administration

(Hard et al. , 1988). Milk also contains many hormones and growth

18

factors; two that have received substantial attention are somatotropin

and insulin-like growth factor-I (IGF-I). Milk concentration of bST

only increase when provocative doses of exogenous bST are

administered to dairy cows (Burton et al. , 1994; Juskevich and Guyer,

1990). In the case of IGF-I, small increases in milk concentrations were

observed in the init ial studies. But as the number of animals and range

of situations were expanded, it became evident that milk IGF-I varied

widely between cows and was affected by many factors (stage of

lactation, environment) and that the use of bST had minimal impact on

the milk concentration of IGF-I (Juskevich and Guyer, 1990; Coll ier et

al. , 1991). Only l imited research has been conducted into the enzymes

in milk in relation to the bST administration in dairy animals. No

signif icant differences in the activit ies of aminopeptidase,

endopeptidase, phosphatase, protease, lipase (Lynch et al. , 1988) and

lipoprotein lipase (Lough et al. , 1989) in milk have been found in milk

from treated and untreated animals. The pH of the milk has also been

found to be not affected by bST injections in different studies (Baer et

al. , 1989). Overall, studies on the macro and micro components of milk

indicate that composit ion is unchanged by use of bST.

2.6 Calving interval and bST

Intercalving interval is often increased as a result of

bSTadminstration’ McGuffey et al. (1991) observed the lactation

performance of multiparous Holstein cows from five herds

supplemented with 0, 320, 640, or 960 mg of Somidobove every 28 d in

early, mid or late stages of lactation. For the early cows, days to

postpartum estrus and service period were similar, however, total

number of inseminations for cows receiving Somidobove was twofold

greater than for the control animals, resulting in a longer calving

19

interval. The duration of treatment with bST also affects intercalving

interval. Esteban et al. (1994) studied the relationship of calving

interval with conception in cows treated for two lactations with

recombinant bST. Multiparous cows treated during the f irst lactation

had signif icantly longer intervals from calving to conception. However,

cows treated during the previous lactation, but not during the second

lactation, had signif icantly shorter intervals from calving to

conception.

2.7 Reproductive performance and bST

The effects of bST on reproductive performance appear to be

variable. Judge et al. (1999) administered bST (500 mg) every 14 d

beginning at 63 d of lactation and continuing unti l 21 d prior to the end

of lactation. They found bST had no signif icant impact on reproductive

performance. Silvia et al. (2002) also found reproductive performance

to be unaffected by bST. In their study cows were randomly allocated

to receive bST supplementation at 14 d intervals, beginning during

weeks 9 to 10 and weeks 17 to 18 after calving. Administration of bST

at weeks 17 to 18 postpartum had no beneficial effect on reproductive

performance of multiparous cows. However, the length of exposure to

bST does not appear to y have an impact on reproductive performance.

Esteban et al. (1994) investigated reproductive performance in cows

exposed for two lactations to recombinant bST. During the f irst

lactation, the chances of a cow becoming pregnant decreased l inearly as

dose increased. During the second lactation, pregnancy rates were

similar to those in the f irst lactation, with again the rates reduced as

bST dose increased.

20

Whilst the hormonal induction of lactation may have an adverse

effect on milk yield in bST treated cows, it does not affect subsequent

reproductive performance of these cows. Mellado et al. (2006) util ized

high-yielding dairy cows in a large dairy operation where bST was

routinely used in all cows. The proportion of cows pregnant was similar

in induced cows and non-treated cows (71 vs. 75%). Thatcher et al.

(2006) reported that coupled with a timed-insemination program or

cows detected in estrus, administration of bST increased embryo

development and embryo survival. In contrast, Waterman et al. (1993)

suggested that chronic administration of a high dose of bST could

reduce reproduction performance by promoting ovarian acyclicity.

2.8 Conception rate and bST

Changes in conception rate in response to the administration of

bST have been variable. Luna-Dominguez et al. (2000) investigated the

effects of bST treatment on reproductive performance in two high

producing herds and found that at 180 d postpartum 65.4 vs. 48.5%

cows were pregnant in control and treated groups, respectively.

However, f irst-service and conception rate were not signif icantly

different between the groups. In contrast, Santos et al. (2004) found

that bST improved conception rates in cyclic cows. Moreira et al.

(2000) examined the effect of bST on pregnancy rates to a t imed

artif icial insemination protocol and to test a resynchronization system

with two consecutive synchronized services. Pregnancy rates were

increased in cows where bST treatment was init iated at 63 ± 3 d

postpartum, compared with cows init iating bST treatment at 105 ± 3 d

postpartum. Combining administration of bST with dietary

manipulation may be beneficial. Billy et al. (2006) examined the

21

effects of bST, pregnancy, and dietary fatty acids on reproductive

responses in lactating dairy cows and found that bST increased milk

production, pregnancy rate (83% vs. 40%) and conceptus length (45 vs.

34 cm). Similarly, Starbuck et al., (2006) found that when lactating

dairy cows, dairy heifers, and lactating beef cows were administered

500 mg bST, artif icial insemination conception rates were improved,

however, the retention of pregnancy was not affected.

Heat stress is often associated with poor reproductive

performance as well as reductions in animal performance and health. To

enable investigation of the effects of bST on reproductive function in

lactating dairy cows during heat stress, Jousan et al. (2007)

presynchronized cows with two injections of PGF2 given 14 d apart

followed by a modif ied Ovsynch protocol. They found that pregnancy

rates did not differ between bST and control cows for f irst and second-

service.

2.9 Dry matter intake and bST

Intake is an important factor that influences nutrient uti l ization,

feed eff iciency and ultimately the profit from the dairy animals.

Vallimont et al. (2001) reported that bST had no effect on DMI or the

digestibi l ity of DM, nitrogen and energy in dairy cows. However,

Santos et al. (1999) demonstrated that cows receiving bST in early

lactation had lower DMI during the f irst 45 d of treatment, whilst

Bauman et al. (1985) and Hartnell et al. (1991) found that with an

extended period of bST administration DMI linearly increased with the

increment of milk production. This increase in DMI occurred within 3

to 5 weeks after bST treatment while the response in milk yield started

within 2 weeks of treatment. This means that initial ly the nutrients for

22

increased milk production must come from body reserves (Peel and

Bauman, 1987) and thus in the f irst few weeks of treatment cows may

experience a period of negative energy balance, depending upon their

energy status at the start of bST treatment, the magnitude of the

production response to bST and the energy density of the ration.

Researchers have suggested various schemes for feeding bST-

treated cows including increasing the amount of grains in feed

(Tessman et al., 1988), feeding a high-energy total-mixed ration ad

libitum (Hutjens, 1990), increasing the amount of energy (Lormore,

1990), increasing the amount of ruminally by passed fat (Chalupa and

Galligan, 1989) or increasing total protein and intake of undegraded

protein (McGuffey et al. , 1991). Every one of these schemes have

revealed very low effects on bST response to intake. However, in a

large f ield study (Thomas et al., 1990), cows were found to respond

well to bST over a wide range of ration nutrient composition.

Preliminary studies have demonstrated that bST responses are greatest

when feed is available for consumption at least 20 hours a day (Patton

and Heald, 1992). These studies showed that when forage quality was

poor, bST response appeared to be reduced. So, overall intake response

to bST can be improved by making the ration more balanced and by

increasing the feeding time.

Intake response to bST, with different dietary energy

concentrations, for the growth performance of bull calves was studied

by Holzer et al. (1999). They found DMI was inversely related to the

energy concentration of the diet and that feed eff iciency was

signif icantly improved by the bST treatment. Peters (1986) also

reported that the level of nutrition may have a large effect on the

response to bST administration.

23

2.10 Dietary energy and bST

Treatment with bST is reported to result in coordination of

metabolism to meet nutrient needs for increased synthesis of milk

components. Bauman et al. (1988) examined the effects of bST on

irreversible loss rate (ILR) and oxidation rate of glucose and

nonesterif ied fatty acids (NEFA) and found that bST treatment

increased milk energy yield by 31% but did not affect feed intake.

Blood glucose concentrations were not affected by treatment; however,

glucose ILR was increased and glucose oxidation was reduced to

accommodate the addit ional glucose (+1.3 mol/d) required for the

increase in lactose secretion. The increases in milk energy yield with

bST treatment caused cows to be in a substantial negative net energy

balance. Other researchers have also investigated the interactions

between bST and dietary energy. McGuffy et al. (1991) reported that

bST increased milk and 3.5% FCM but had effect on DMI and net

energy intake.

Nutritional status of the animal has an effect on the response to

bST treatment. Austin et al. (1991) studied the interactions of bST and

nutrient density during weeks 5 to 20 postpartum. Administration of

bST increased milk production, but augmentation had correlation with

dietary energy and protein. Percentage of milk fat was not affected by

treatment; however, milk protein percentage was reduced with the high

protein and energy diet Kirchgessner et al. (1991) also evaluated

energy metabolism in bST-treated lactating cows and found that

mobil ization of body energy compensated for the bST-induced increase

of milk energy and supported milk production of underfed cows by two-

thirds of the net energy deficit. The uti l ization of mobilized body

energy and ME from the diet for maintenance and milk production was

24

unchanged. The bST and energy deficiency increased N uti l ization for

milk production, because bST increased milk N secretion at the same N

intake, and energy deficiency partly compensated the reduced N intake

by N mobilization.

Administration of bST has variable effects on nutrient uptake.

Miller et al. (1991) reported a potential fourfold increase in NEFA

uptake in bST-treated cows while Tessman et al. (1991) found that

daily injection of bST did not change blood concentrations of glucose

or urea or plasma β-hydroxybutyrate concentrations. Similarly,

McGuire et al. (1992) investigated the relationship bST and dietary

energy and protein status in mid-lactation Holstein cows and found

that milk yield increased for cows fed all diets with bST, but the

response was greatest for those fed the high NE/high CP diet. For all

diets, bST caused an increase in plasma IGF-I (125%) and IGF-II

(21%), with the increase being substantially greater for cows fed the

high NE/high CP diet. Basal insulin levels differed among diets and

increased with exogenous bST in cows fed the high NE and high CP

diet. Leonard and Block (1997) studied the effects of bovine

somatropin on nutrients util ization with either a glucose or insulin

infusion, with their results suggesting that, in early lactation, insulin

was sti l l anabolic because the body weight gain of cows increased.

However, milk yield was stil l higher than that for cows in late lactation

with similar insulin concentrations.

2.11 Dry matter digestibil ity and bST

In a trial on 24 dairy cows, Santos et al. (1999) determined the

digestibi l it ies of DM, organic matter (OM), crude protein (CP), starch,

neutral detergent f iber (NDF) and acid detergent f iber (ADF) in bST

treated animals. Treating cows with bST had no effect on the

25

digestibi l it ies of DM, carbon, and energy, confirming the previous

results of Robinson et al. (1991) who showed that basal metabolism and

maintenance requirements are unaffected by bST treatment. Similarly,

when calculating nitrogen balance, bST administration did not alter

feed digestibi l i ty. In contrast, nutrient demand and metabolism may be

altered by bST administration, but these alterations are attributed to the

animal factors l ike body weight, milk production, milk fat test, and

expected gain or loss of body weight (NRC, 1988).

Santos et al. (1999), while studying the effect of bST with two

types of diets containing corn and sorghum, concluded that bST had no

effect on digestibi l it ies of DM, NDF, ADF, CP and OM for either diet,

and furthermore, no interactions between diets and bST treatment were

observed. Other studies (McGuffey and Wilkinson. 1991) also revealed

no effects of bST on apparent digestibi l i ty of nutrients. However,

Newbold et al. (1997) demonstrated that supply of fermentable energy

may mediate response to bST by increasing the negative energy balance

observed during early lactation. This indicates that digestibi l i t ies of

different nutrients can be increased by providing readily available

source of energy l ike molasses.

2.12 Feed efficiency and bST

The eff iciency of feed util ization is often increased in response to

administration of bST. Bauman et al. (1985) found that when Holstein

cows were treated with either 13.5, 27, or 40.5 mg bST/d average FCM

yield increased in a dose-dependent fashion from 23 to 41%. Overall,

gross lactational eff iciency (milk per unit of net energy intake) was

improved by treatment with bST. Similar results were also obtained by

Solderhalm et al. (1988), who treated Holstein cows with subcutaneous

26

injections of 10.3, 20.6, or 41.2 mg bST daily. Cows treated with bST

consumed 4-10% more feed, but were 11-17% more eff icient than the

controls for conversion of feed to milk. Huber et al. (1997) found that

milk yield and eff iciency of feed uti l ization were enhanced in cows

injected with bST for four consecutive lactations. Santos et al. (1999)

also found that milk yield and efficiency of feed uti l ization were increased

with bST. Similarly, Remond et al. (1991) investigated the effects of

bST (500 mg injected at 14d intervals) with varying level of energy

concentrate in the diet and found that milk yield of bST injected cows

increased 11.0% and that gross eff iciency of milk yield was improved

2.13 Animal health and bST

Treatment of cows with bST is generally associated with no

adverse effects on animal health, although there are some instances

where this may not be the case. Lean et al. (1991) studied post

parturient metabolic disorders and production responses in animals that

had been previously exposed to long-term treatment with bST and

concluded that bST use in a prior lactation may have potential benefit

in reducing risk of metabolic disorders associated with l ipid

mobil ization in the post parturient period.

The effect of bST on the incidence of mastit is appears to be

variable. White et al. (1994) reported that the incidence of clinical

mastitis within a herd was similar, regardless of whether or not the cows

were treated with bST. Zhao et al. (1992) found that mostly the health

and reproduction related variables were not affected by bST treatment,

although the incidence of teat and udder disorders and feet and leg

problems tended to be higher in bST-treated cows. Lack of effect of

bST on the incidence of mastitis was also reported by Collier et al.

27

(2001) who found that bST treatment had no effect on total mastitis

cases, total days of mastit is, duration of mastit is, or the odds ratio of a

cow to develop mastit is. In contrast, Hoeben et al . (1999) found that

bST had a protective effect during experimental Streptococcus uberis

mastitis in cows, whilst Dohoo et al. (2003) found bST increased the

risk of clinical mastitis by approximately 25% and increased the

chances of developing clinical signs of lameness by 55% during the

treatment period.

2.14 Blood metabolites and bST

Treatment of cows with bST affects various blood metabolites.

Guillermo et al, (1990) found blood pH and buffering capacity

decreased as a result of long-term administration of sustained-release

bST. The decrease in buffering capacity was due to decreased

bicarbonate without affecting blood partial pressures of oxygen or

carbon dioxide. Concentration of plasma albumin was decreased and

NEFA, glucose, insulin, bST and IGF-I was increased by treatment.

Total plasma proteins and cortisol, hemoglobin, and hematocrit were

unaffected. Liver DM, total l ipid and triacylglycerol contents, and

plasma 3-hydroxybutyrate and glutamic oxalacetic transaminase were

not affected by bST, but the percentage of triacylglycerol in total l ipids

was increased. These results suggest that bST exhibits lipolytic

activit ies in lactating dairy cows. Responses in blood metabolites may

vary depending on the parity of the cow and the level of bST treatment.

Lean et al. (1992) found that cows treated with 51.6 or 86 mg bST/d

had signif icantly higher serum-free fatty acids than controls. They also

found that older cows treated with 86 mg of bST/d tended to have

higher blood glucose concentrations than control-group cows.

28

Other researchers have also reported parity effects to bST

administration. Skarda et al. (1992) found that administration of bST

resulted in elevated plasma bST during the f irst 9 d after injection;

however, blood erythrocyte and leucocyte counts, haemoglobin

concentrations, haematocrit values, plasma levels of glucose, FFA,

urea, P, serum alanine aminotransferase and aspartate aminotransferase

were not affected. Mishra and Shukla, (2004) in their study with

lactating Murah buffaloes, reported that glucose, triglycerides, total

proteins, albumin, globulin and electrolytes l ike sodium and potassium

were not altered by bST. However, they observed a signif icant decrease

in blood urea nitrogen (BUN) concentration in response to bST

treatment.

Plasma IGF-I concentrations are affected by bST treatment. In

studies involving a prolonged-release formulation of sometribove (N-

methionyl bST) Schams et al. (1991) found that plasma ST increased

signif icantly at d 7 after bST treatment but then decreased on d 14. In

addit ion, plasma IGF-I was posit ively correlated with ST. Rose et al.

(2004) found plasma IGF-I concentrations following injections of bST

was greater low response (LR) cows, whereas the difference in plasma

IGF-I level between week 1 and 3 was greater for the high response

(HR) cows due to lower levels of 3-hydroxy-butyrate and NEFA before

bST treatment. Chaiyabutr et al. (2005) also found that bST increased

the concentration of plasma IGF-I throughout the lactation period,

while plasma glucose, protein and triglyceride concentrations remained

stable. Slaba et al. (2005) measured plasma bST and IGF-I

concentrations in dairy cows following a single subcutaneous injection

of a slowly released preparation of either recombinant enterokinase

linker bST or recombinant methionyl bST. There was a three to seven

29

fold increase in plasma bST concentrations during the f irst three hours

post-injection in cows treated with both sometribove (from 3.4 ± 0.8 to

11.2 ± 3.0 ng/ml) and somidobove (from 2.3 ± 0.3 to 17.5 ± 2.6 ng/ml).

In the next 8 d the bST concentration in the bST-treated cows varied,

but was stil l signif icantly higher than the controls. In the following

days, the concentrations of bST did not differ from the controls.

2.15 Economics and bST

There are variable reports on the f inancial benefit of bST for milk

production. Tauer and Knoblauch, (1997) using data from 259 dairy

farm business in the USA, found that, on average, farms using bST had

25% longer lactation length, produced 510 kg more milk per cow and

net farm income increased by $27 per cow as compared to the farms

where bST was not used. Kinoshita et al. (2004) developed a

comprehensive economic model to evaluate potential impacts of bST in

Japan and concluded that bST can have negative effects in smaller

farms as compared to large farms due to higher costs, and lower profit-

earning ability. Judge et al. (2009) evaluated the economics of four

dairy farms and reported that the overall average change in net farm

income (NFI) attributable to bST was $43.01 per bST-treated cow.

However, profitability of bST use was observed to be quite variable

between farms studied due to the level of production response and the

price received for milk. Similarly, Tauer (2008) using data over the

years 1994 through 2004 for selected New York dairy farms found that

bST increased milk production per cow and decreased the cost of

production per hundredweight of milk.. Mc Bride (2002) assessed the

production and financial impacts of bST adopted U.S. dairy operations

and reported that a substantial increase in milk production per cow is

associated with bST adoption, but large estimated financial impacts

30

were not statistically signif icant because the substantial variation in the

net returns of bST adopters may be related to the management-intensive

nature of bST.

31

CHAPTER -3

Effect of long term use of bovine somatotropic hormone on milk production, reproduction and blood metabolites in Nili- Ravi

buffaloes

ABSTRACT

The objectives of the study were to investigate the effects of

administering bST on milk production, milk composition, reproduction

and hematological and biochemical parameters in Nili-Ravi buffaloes

for a period of three years, from 2004 to 2007. Thirty (n = 30) Nili-

Ravi lactating buffaloes with similar milk production and stage of

lactation were selected and randomly divided in to two groups, A and

B, with 15 animals in each group. Group A (0 bST) served as the

control while animals in group B (+bST) were injected every 14 d with

Boostin-250™ (250mg/animal). Nutritional requirements of the

experimental animals were met through available green fodder (45-50

kg/d) supplemented with concentrate ration at a rate equivalent to half

of the milk production of the animal. Average daily milk production

per animal was 6.76 vs. 7.98 kg for control and treated groups,

respectively. Milk production increased by 18.04% in response to bST

treatment. Numerical variations in the values of the percentages of fat,

SNF, and TS were observed, although the variation of these parameters

in both the groups was non signif icant. The calving interval, dry period

and lactation length were shorter by 14.0, 26.0 and 2.7%, respectively

in the treated group compared with the control. The postpartum estrous

period, service period and services per conception were 98.2 vs. 160 d,

115.10 vs. 207.04 d and 1.31 vs. 1.47 in the bST treated and control

groups, respectively. Statistically, differences were signif icant for

postpartum estrous and service period but for services per conception

32

the difference was non-signif icant. Results indicated positive effect of

bST on reproductive parameters. Prevalence of mastitis was 10.0%

higher in bST-treated animals. Although there were variations in body

weights for animals in groups A and B, these changes over the time of

the study were non-signif icant. The differences among hematological

and biochemical parameters were also non-signif icant except blood

urea nitrogen (p< 0.05), which was higher in the control group. The

economic benefit over a 305 d lactation period with the use of bST was

(Rs. 4227.0).

INTRODUCTION

Pakistan has approximately 27.33 mill ion buffaloes, which

contribute more than 73% of total milk production and 42% of beef

production in the country (Ministry of Food and Economics and

Statistics, 2006). In spite of this large number, per animal productivity

is low as compared with western dairy cattle breeds. The average milk

production is 7-8 kg/d at farm level, which needs to be increased to

make this animal more profitable as well as to meet the protein

requirements of the ever-increasing human population in the country.

Breeding and genetic selection failed to increase much the amount of

milk produced by buffaloes as did for cows (Otto et al., 2003). The

biotechnological treatment (bST) supported by appropriate nutrit ion

and herd management can be a feasible substitute for high buffalo milk

production (Ludri et al. , 1989).

Administration of bST is proved to increase milk yield without

marked changes in composition in dairy animals (Bauman et al., 1985;

Rose et al., 2004; Van Bbaale et al., 2005). It improves the productive

eff iciency due to increased milk yield per unit of feed intake (Bauman,

33

1992). It has no effect on reproductive performance, but tended to

improve some measures of reproduction (Judge et al. , 1999; Silvia et

al. , 2002). Increased milk production in dairy cows with bST has been

shown to be cost effective and without any adverse effects on animal

health (Bauman, 1992).

Little information is available on the use of bST in buffalo, with

only a few studies of short-term duration having been carried out. The

present study was undertaken to investigate the effect of long term use

of bST on productive, reproductive, health, physiological and

biochemical parameters in Nil i-Ravi buffaloes.

MATERIALS AND METHODS

Experimental design

The trial was conducted to study the effect of administration of

bST in Nili-Ravi buffaloes on productive, reproductive, physiological

and biochemical parameters for a period of three years. Thirty (30)

multiparous Nili-Ravi buffaloes with similar stage of lactation and

parity were selected from the herd of the Livestock Experiment Station

Directorate of Livestock Production Research Institute, Bhadurnagar,

and Okara. The average milk yield of the experimental animals was 08

± 1 kg/d and average body weight was 540 ± 15 kg. The animals were

randomly divided in to two groups, A and B, with 15 animals in each

group. Group A was kept as control, while group B was exogenously

injected with the (bST) Boostin -250™ (manufactured by LG Life

Sciences, Ltd. South Korea and distributed by M/S Ghazi Brothers,

Pakistan). The dose level was 250 mg/ animal as recommended by the

manufacturer and subcutaneous injection was given at fortnightly

34

interval after 60 ± 3 d postpartum until the end of lactation. The same

pattern of bST administration was followed in the subsequent

lactations.

Experimental ration

The nutrit ional requirements of experimental animals were

fulf i l led according to NRC (2001) on the basis of body weight and milk

production through the provision of green fodder and concentrates.

Maintenance requirements were largely met through available green

fodder. Winter fodders were chopped berseem (Trifolium pretense) and

lucerne (Medicago sativa) mixed with wheat straw, while summer

fodders were chopped maize (Zea mays), sorghum (Sorghum bicolor)

and sadabehar (Sorghum bicolor x Sudanense). The average chemical

composition of these fodders is presented in Table 1. The daily

allowance of 50 ± 5 kg fodders/d was provided to each (individual)

animal. The milk production requirements were met through provision

of the concentrate ration, which contained 17.39% CP and 2.45 Mcal

ME/kg on DM basis. Concentrate was offered individually at the time

of milking on the basis of daily milk production. The composit ion of

concentrate ration is given in Table 2.

Feeding program

Chaffed, green fodder was fed to the animals in the shed while

the concentrate mixture was offered at the time of milking. The

concentrate was fed at a rate equivalent to half of their average weekly

milk production. Samples of feed and green fodder were collected at

fortnightly interval and analyzed for proximate analysis according to

AOAC (1995). The gross energy of the ration was determined through

the IKA C-2000 Bomb Calorimeter, while metabolizable energy was

35

calculated as 63% of the gross energy (Mandal et al ., 2003). The

analysis was done in the Nutrition Laboratories of the Department of

Food and Nutrit ion, University of Veterinary and Animal Sciences,

Lahore.

Management of animals

During the experimental period, homogeneous management

practices were adopted for al l animals. They were housed in the same

shed equipped with shade, mangers and water troughs; the shed was

partitioned to accommodate both groups separately. The vaccination

schedule was followed against contagious diseases (especially

Hemorrhagic septicemia and Foot and Mouth Disease) according to

farm routine, whereas for the control of parasitic infestation, animals

under tr ial were dewormed with a broad-spectrum anthelminthic

(Nilzan plus™) twice a year.

The fol lowing parameters were studied to see the influence of bST.

1. Milk production and composition

Buffaloes were hand milked twice daily at 02.00 and 14.00 h and

milk production at each milking was weighed and recorded. Daily milk

recording commenced 3 d post-parturition and continued until the

completion of lactation. Milk was sampled from consecutive a.m and

p.m milking each week and composited for analysis. Milk samples were

used to determine fat %, and SNF %, using the Gerber and Rapid

methods, respectively as described by FAO (1986a and 1986b). Total

solids % was determined by the Gravimetric method according to

AOAC (1990).

36

Table 1

Average chemical composition of fodders Fodders DM

(%) CP (%)

NDF (%)

ADF %

ME (Mcal/kg)

Ca (%)

P (%)

Winter Fodders

Berseem (Trifolium

pretense) 20.23 18.82 51.32 41.97 2.17 1.66 0.33

Lucerne (Medicago

sativa) 24.95 21.25 52.67 40.59 2.24 2.00 0.32

Wheat straw 92.75 2.59 80.9 50.2 1.55 0.30 0.07

Summer Fodder

Maize (Zea mays) 25.93 7.20 66.01 34.11 2.0 0.52 0.28

Sorghum (Sorghum

bicolor) 28.8 6.86 67.09 37.90 1.86 0.58 0.12

Sadabahar

(Sorghum bicolor x

Sudanese)

24.16 9.70 65.06 41.64 1.79 0.65 0.17

DM = dry matter, CP = crude protein, NDF = neutral detergent f iber, ADF = acid detergent f iber, ME = metabolisable energy, Ca = calcium, P = phosphorus

37

Table 2 Composition of concentrate ration Ingredients Percentage

Maize grain 10.00

Rice polishing 8.00

Wheat bran 22.00

Cotton seed cake 14.00

Canola meal 10.00

Maize gluten meal 30 % 22.00

Molasses 12.00

Mineral mixture* 2.00

Chemical composition

Dry matter (%) 91.00

Crude protein % 17.39

Metabolisable energy (Mcal/kg) 2.45

Neutral detergent f iber (%) 32.17

Acid detergent f iber (%) 18.30

Calcium (%) 0.70

Phosphorus % 0.45

* Mineral mixture composition (/kg) Dicalcium phosphate 708 g; Sodium choloride 189 g; Magnassium sulphate 86.0 g; Ferous sulphate 8.9 g; Manganese sulphate 4.9 g; Zinc sulphate 3.2 g; Copper sulphate 0.3 g; Potassium iodide 0.087 mg and Cobalt chloride 0.0089 mg ; Sodium selenate 0.015 mg.

38

2. Reproductive parameters

During the experiment the animals were closely observed for

response in reproductive parameters under the influence of bST

hormone.

2.1 Postpartum estrous

The exhibition of heat (estrus) in the cows was detected through

twice daily (2 h) exposure to vasectomised bulls at 5.00 and 17.00 h.

Observations on the onset of estrous were recorded for individual

animals in both of the groups.

2.2 Services per conception

The animals were inseminated using artif icial insemination

technique. The number of services for establishing pregnancy were

recorded for each animal.

3. Health data

The animals were watched closely for any ailment (infectious and

metabolic) and any incidences were recorded. The animals were also

monitored for udder health by carrying out the sub-clinical mastitis test

(Muhammad et al., 1995) on a weekly basis.

4. Physiological parameters

Blood samples were collected every 14 d by puncturing of the

jugular vein and collecting blood in two (10×100mm) evacuated tubes;

one containing sodium heparin and the other without anticoagulant. For

those animals in Group B the blood was collected prior to the bST

injection. Immediately fol lowing collection, the blood samples were

39

placed on ice. The collection tubes containing 0.2 ml sodium heparin

were used for determination of erythrocyte sedimentation rate with

wstertergen sedimentatation tubes followed by Benjamin, (1985). The

Pack cell volume was noted as percentage with the help of hematocrit

scale explained by Bush, (1991). While, the haemoglobin contents of

the plasma were monitored through Sahili ’s Apparatus described by

Sastry, (1983). However, the tubes without anticoagulant were allowed

to clot, centrifuged at 3000 RPM for 30 min. at 5°C and then the

harvested serum was stored in capped glass bottles at -4°C til l analysis.

For determination of blood chemistry, preserved samples of serum were

analyzed by using Cecil 2041 single beam spectrophotometer with

Randox Kits developed by Randox Laboratories Ltd. United Kingdom. However, non-metal elements like sodium and potassium were

determined through Sherwood flame photometer-410 in the laboratories

at University of Veterinary and Animal Sciences, Lahore.

5. Statistical Analysis

Data on productive parameters (milk production, lactation length,

dry period and calving interval), milk composition (fat, SNF, TS),

reproductive parameters (postpartum estrus, service period and services

per conception) and blood metabolites (blood glucose, blood urea

nitrogen, total protein, cholesterol, total l ipids, calcium, sodium,

potassium, Hemoglobin, ESR and PCV) were statistically analyzed by

student’s t-test using MINITAB® software package of statistical

analysis.

40

RESULTS AND DISCUSSION

Milk production

The weekly milk production pattern in both the groups during

the f irst year of the study is shown in Figure 1. During this f irst year

the average daily milk yield of groups A and B was 6.57 and 7.76 kg,

respectively (Table 3), representing an increase (P<0.05) in milk

production of 18.11% in response to bST treatment. These findings

are agreement with those of Jorge et al. (2002), Mishra and Shukla,

(2004) who found that the increase was 14.93 to 32.4% in treated

Murrah and Kundi buffaloes, respectively, although the increase

depended on the dose rate of bST.

During the second year of the study the average daily milk

yield of groups A (control) and B (treated) was 7.64 and 8.09 kg,

respectively (Table 3). The weekly milk production pattern in both the

groups is given in Figure 2. The increase in milk production in the

treated group (B) over the control (A) was 5.90 %, which was much

less than the response in the f irst lactation, and furthermore the

difference observed in the second year was non signif icant (P>0.05).

This response to bST in the second year may have been confounded due

a number of animals contracting FMD. Given that more animals (11) in

the bST-treatment group (group B) were infected than in the control

group (8), this would have had a greater impact on average milk yields

of the bST-treated group. This argument for the lower response in the

second year is supported by Bauman, (1992) who reported that poorly

managed farms, where animals are stressed, underfed or sick will have

achieved negligible milk response to bST. However, it is also possible

that the lower response to bST in the second year was not due to the

41

health status of the animals as Hansen et al. (1994) found the increase

in milk production during the second year (of bST treatment) was

decreased by an amount equivalent to 67% of the total increase

observed in the f irst year of use of bST in dairy cows.

The average daily milk yield for the third consecutive year was

5.97 and 8.11 kg for group A and B, respectively (Table 3),

representing a 35.84% increase (P<0.05) in milk yield in response to

bST. The weekly milk production trend of both experimental groups is

reflected in Figure 3, showing a similar trend as observed in the f irst

and second years. The increase in milk yield of the bST-treated animals

may not have been due entirely to bST as some of the animals within

this group were parity 3 (peak lactation) whilst all animals in the

control group were parity at declined lactation. However, the results of

the present study were consistent with those of Jorge et al. (2000), Nisa

et al. (2006) and Sarwar et al. (2007) who reported that bST

administration in buffaloes increased milk production from 30 to 37%.

Over the entire trial period, the average daily milk production

was 6.76 vs. 7.98 kg in control and treated group, respectively (Table

3, Figure 4); representing an increase (P<0.05) of 18.04% in response

to bST treatment. Similar results have been reported in previous

studies. Huber et al. (1997) reported that milk production was

increased by 14% in dairy cows injected every 14 d with 500 mg bST

for four consecutive lactations. Hemken et al. (1991) also reported

that milk production was increased in dairy cows receiving bST for

two consecutive lactations. The increment in milk production may be

that bST is a homeorhetic controller that shifts the portioning of

nutrients so that more are used for milk synthesis (Peel and Bauman,

1987). Thus, effects are primarily, perhaps exclusively, on directing

42

the use of absorbed nutrients. This involves coordinating the

metabolism of various body organs and tissues; these orchestrated

changes in tissue metabolism involve both direct effect on some tissue

and indirect effects mediated by somatotropic hormone dependent

somatomedians (IGF-I) for other tissues like mammary glands

(Bauman, 1992). The mechanism of bST to exert its galactopoietic

effect is complex and involves a multiplicity of events in the whole

animal. Increases in the availability of glucose with in the mammary

epithelial cell in response to growth hormone treatment would result

in increases in rate of lactose synthesis and hence stimulation of milk

production. Increases in the concentration of IGF-I in milk and plasma

could be one of the processes involved in stimulation of milk

synthesis (Faulkner, 1999). In addition, bST appears to promote milk

production by a partitioning effect on absorbed nutrients so as to

supply more substances for mammary synthesis and also the level of

nutrition may influence yield responses for milk and nutrient f low

(Bauman and Currie, 1980). The increase in milk synthesis most likely

involves changes in the activity of key regulatory enzymes resulting

in an increased synthesis rate per epithelial cell.

Lactation length

The average lactation length of the treated (+bST) and control

(0 bST) groups were 259.4 vs.264.0, 256.2 vs. 275 and 251.0 vs. 0 d

for lactations 1, 2 and 3, respectively (Table 4). Relatively, the

lactation length of the control group was longer to that of treated

(+bST) group, although this difference was not statistically signif icant

(P>0.05). No animal in the control (0bST) group calved and

completed their 3rd lactation during study period, while five out of fifteen

43

(5 /15, 33.3%) of the bST-treated group calved and completed their 3rd

lactation.

The overall mean lactation length group for treated (+bST) and

control (0 bST) groups was 253.0 and 260.0 d, respectively (Table 5

and Fig. 5). However, this difference was not statistically signif icant

(P>0.05). This indicates that bST does not have negative effect on

productive traits in buffaloes, which agrees with the results of

Bauman et al. (1999) and Jones (2000) who reported that rbST

treatment with good management had no negative effect on productive

and fertil ity traits in cattle.

0

2

4

6

8

10

12

1 3 5 7 9 11 13 15 17 19 21 23 25 27

Weeks

Milk

yie

ld (

kg/d

)

Treated

Control

Figure 1. Trend of milk production in control and bST-treated groups

during the f irst year of the study.

44

0

2

4

6

8

10

12

1 4 7 10 13 16 19 22 25 28 31 34 37 40

Weeks

Milk

yie

ld (

kg/d

)

Treated

Contol

Figure 2. Milk production pattern in the control and bST-treated

groups during the 2nd year of the study.

0

2

4

6

8

10

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35

Weeks

Milk

yie

ld (

kg/d

)

Treated

Control

Figure 3 Trend of milk production in the control and bST-treated

groups during the 3rd year of the study.

45

0

2

4

6

8

10

12

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39

Weeks

Milk

yie

ld (

kg/d

)

Treated

Control

Figure 4. Comparison of milk production in control and bST-treated

groups over the 3 year study period.

Table 3.

Effect of bovine somatotropic hormone administration on average milk yield of Nili- Ravi buffaloes (means ± SE)

Parameters

Study period

Group A (0 bST)

Group B (+bST)

Yield SE Yield SE

Milk

production

(kg/day)

Year-1 6.57 a 0.29 7.76 b 0.33

Year-2 7.64 a 0.33 8.09 a 0.31

Year-3 5.97 a 0.23 8.11 b 0.20

Overall 6.76 a 0.25 7.98 b 0.26

Means within row bearing different superscripts differ significantly (P<0.05). SE = Standard error bST= Recombinant bovine somatotropic hormone.

46

Dry period

The average length of the dry period following the f irst lactation

was 204.2 vs.246 d for the bST-treated and control groups,

respectively. The average length of the dry period for the bST-treated

group during the second lactation was 160 d (Table 4). No animal in

the control (0bST) group calved and thus none completed their 3rd

lactation during the study period. The overall average length of the

dry period in treated and control groups were 182.1 vs. 246.0 d,

respectively (Table 4), although these values were not signif icantly

different (P>0.05). The trend in length of the dry period is shown in

Figure 5.

Calving interval

Between the f irst and second lactations the average calving

interval of bST-treated and control groups were 460 and 506 d,

respectively, although these values did not differ signif icantly

(P>0.05). The intercalving interval between the second and third

lactations could only be determined for the bST-treated group and

which was 410 d. Despite being unable to determine the average

intercalving interval of the control group over the entire study period,

the fact that the control group animals did not calve in the f inal year

of the study is the basis for concluding that bST-treatment resulted in

a decrease in the intercalving interval. The relative shorter period may

be due to improvement in fertil ity because higher peak in

progesterone production has been reported in the f irst two cycles of

bST treated cows and it was suggested that this may have been due to

ovarian IGF-I mechanism (Gallo and Block, 1990). The results of the

present study are in agreement with those of Thomas et al. (1990) and

Jones (2000) who reported that bST treatment with good management

47

has no negative effect on productive and fertil ity traits in cattle. In

the study presented here, bST-treatment had a positive effect on

fertil ity traits by decreasing the intercalving interval.

Table 4

Effect of recombinant bovine somatotropic hormone on productive traits of Nili- Ravi buffaloes (mean ± SE)

Parameters

Parity

Control (0 bST)

Treated (+ bST)

Days SE Days SE Lactation

persistency

Dry period Calving interval

Lactation -1

Lactation -2 Lactation –3 Overall average Lactation -1 Lactation -2 Overall Average

Lactation -1 Lactation -2 Overall average

264.1

275.1

--

260.0

246.0

--

246.0

506.0

--

506.0

15.0

12.0

---

9.5

34.0

--

34.0

33.0

---

30.0

259.4

256.2

251.0

253.0

204.2

160.0

182.1

460.0

410.0

435.0

8.8

13.0

4.2

6.0

27.0

22.0

20.0

30.0

20.0

23.0

SE= standard error. bST= Recombinant bovine somatotropic hormone.

48

Figure 5. Comparison of lactation length (LP), dry period (DP) and calving interval (CI) in bST-treated and control groups during the study period

Milk composition

Administration of bST had no effect (P>0.05) on any of the

measures of milk components. The average milk composition over the

study period were 6.79 vs. 6.75% fat, 9.36 vs. 9.27% SNF, and 16.15

vs. 16.00% TS for bST-treated and control groups, respectively (Table

5). A comparison of milk components over the study period is shown in

Figures 6, 7 and 8). Linn (1987) observed variations in bST production

trials and reported that there can be minor changes, primarily in fat

content of milk, during the f irst few weeks of bST administration due to

adjustments in the cow’s metabolism and voluntary feed intake.

However, these changes only temporary and negligible when compared

with variations normally occurring over a lactation cycle. Other

researchers, for example, Barbano et al. (1992), Jorge et al. (2002),

Chalupa et al. (1996), Mishra and Shukla (2004), and Khattab et al.

49

(2008) also found that bST did not influence the milk composition of

dairy cows, Murrah, Kundi and Egyptian buffaloes, respectively.

However, Huber et al. (1997) reported that fat percent was decreased in

dairy cows with long term use of bST. Such differences may be due to

variation in the quality and quantity of feed offered to the animals

under trial. The results of the present study are also different from

those of Eppard et al. (1985), Mc Bride et al. (1989) and Capuco et al.

(2001) who showed that milk fat percentage increased in response to

bST treatment. An increase in fat content of milk during bST

administration is often caused by an extra mobilization of the fat

reserves of the animal. Greater increases in fat percent with bST

administration have been observed when cows were in negative energy

balance (Peel et al., 1989). Similarly the results of McGuffey et al.

(1990) are also not in agreement with the present findings as they

observed a signif icant decrease of TS in milk obtained from bST-

treated animals.

50

Table 5.

Effect of bovine somatotropic hormone administration on milk components of lactating Nili–Ravi buffaloes over the 3 year study period (mean ± SE)

Parameters Study period

Control ( 0 bST)

Treated ( + bST)

Percent SE Percent SE Milk components Fat Solid not fat (SNF)

Total solids (TS)

Year -1 Year-2 Year-3 Over all Year -1 Year-2 Year-3 Over all Year -1 Year-2 Year-3 Over all

6.54

6.79

6.89

6.75

9.25

9.28

9.29

9.27

15.79

16.04

16.18

16.00

0.04

0.06

0.08

0.05

0.04

0.06

0.03

0.01

0.08

0.08

0.12

0.05

6.66

6.87

6.83

6.79

9.36

9.38

9.34

9.36

16.04

16.25

16.17

16.15

0.05

0.07

0.07

0.06

0.03

0.06

0.06

0.04

0.07

0.12

0.11

0.09 SE= standard error. bST= Recombinant bovine somatotropic hormone.

51

Figure 6 Comparison of yearly and overall milk fat % of bST-treated

and control groups over the study period

Figure 7 Comparison of yearly and overall milk solid not fat % of bST-treated and

control groups during the study period.

9.15

9.2

9.25

9.3

9.35

9.4

Year -1 Year-2 Year-3 Over all

Study period

Per

cen

t

Treated

control

9.15

9.2

9.25

9.3

9.35

9.4

Year -1 Year-2 Year-3 Over all

Study period

Per

cen

t

Treated

control

52

Figure 8 Comparison of year wise and overall milk total solids % of

bST-treated and control groups during the study period.

Reproductive performance

The reproductive parameters in terms of postpartum estrus days,

service period and services per conception were studied for three

consecutive years to know the long-term effect of bST on fertil ity of

buffaloes. The postpartum estrus days were 139.0, 103.6 and 98.20,

service period was 156, 126.5 and 62.58 d and services per conception

were 1.26, 1.46 and 1.2, respectively in the bST-treated group for

lactations 1, 2 and 3. For the control group the postpartum estrus days

were 175.0 and 239.08, the service period was 175.0 and 239.08 d and

services per conceptions were 1.73 and 1.21 for lactations 1 and 2,

respectively (Table 6). Within the control group of animals, none

calved for their third lactation during the three year study period, while

in the bST-treated group all completed their 2nd lactation and 33% of

15.5

15.6

15.7

15.8

15.9

16

16.1

16.2

16.3

Year -1 Year-2 Year-3 Over all

Study period

Per

cen

t

Treated

control

53

the animals calved for their 3rd lactation. The overall means of

postpartum estrus were 98.20 vs. 160.0 days, service period 115.0

vs.207.04 and services per conception 1.31 vs. 1.47 for the bST-treated

and control groups, respectively. The improvement was observed in

treated group over the control. The statistical analysis revealed that

there was signif icant improvement (P<0.05) in postpartum estrus and

service period in treated group over the control. However, the

improvement was non signif icant (P>0.05) in services per conception

for experimental animals. The results of the study are supported by the

results of Spicer et al. (1990).who reported that granola cells in the

bovine ovary apparently produce IGF-I that acts synergistically with

FSH to increase the production of estrogen and progesterone hwhich

have ultimately effect on reproductive performance in dairy cows.

Health parameters

The effect of long-term administration of bST on the health of

Nili–Ravi buffaloes was monitored through observation of the

incidences of both contagious and non-contagious diseases. The

incidences of mastitis for bST-treated and control group were 5.0 vs.

2.0, 9.0 vs.7.0 and 4.0 vs.5.0 in years 1, 2 and 3 respectively (Table 7).

The increase of mastitis incidences in treated group over the control

was 28.47%., although this increase was non signif icant (P>0.05).

These findings are in agreement with those of White et al. (1994) and

Collier et al. (2001) who found rbST did not increase the risk of

clinical mastitis; instead they found but a positive linear relationship

existed between milk yield and mastitis incidence.

54

Table 6.

Effect of bovine somatotropic hormone administration on reproductive parameters of lactating Nili–Ravi buffaloes during the study period

Parameters

Parity

Control (bST 0)

Treated (bST+)

Days SE Days SE Post partum

estrus

Service period/open

day

Number of services per conception

(Nos.)

Lactation-1

Lactation -2

Lactation-3

Over all

Lactation-1

Lactation -2

Lactation-3

Over all

Lactation-1

Lactation -2

Lactation-3

Over all

145.00

175.00

--

160.00

175.00

239.00

---

207.04

1.73

1.21

---

1.47

11.00

14.00

9.30

22.00

23.00 -

17.00

0.03

0.21

--

0.2

139.00

103.06

52.00

98.20

156.00

126.50

62.80

115.10

1.26

1.46

1.24

1.31

11.00

11.00

14.00

8.30

19.00

18.00

6.50

12.00

0.17

0.50

0.40

0.09

Means with in row bearing different superscripts differ significantly (P<0.05). SE= standard error. bST= Recombinant bovine somatotropic hormone.

55

Figure 9 Comparison of reproduction parameters (post partum estrus [PPE], service period [SP]) of bST-treated and control groups during the study period.

The overall incidences of Foot and Mouth disease were 11.0 vs. 10.0

for the bST-treated and control groups, respectively (Table 7). The

difference in the incidences was non signif icant (P> 0.05) in treated and the

control groups. Only one incidence of lameness was observed during the

study in each group. The lameness was may be the after effect of FMD. On

the overall, the results are in agreement with Burton et al. (1990) who

reported no signif icant difference in the incidence of lameness or hoof

injuries in bST-treated animals.

56

Blood metabolites

During the study period, serum from blood samples collected

from both groups of animals was analyzed for hematology, nutrient

metabolites and mineral contents.

Hematological parameters

The overall (3 year) average hematological parameters for ESR,

HCV and Hb content of control and bST-treated groups were 70.50 vs.

70.70 mm/Ist h, 35.10 vs. 34.55% and 13.00 vs. 13.26 g/dl, respectively

(Table 8). Statistically, al l differences were non signif icant (P>0.05).

These results are similar to those of Mishra and Shukla (2004) who

reported that bST had no effect on hemolymphatic (blood) system of

dairy buffaloes. In contrast, Rowlands et al. (1979) observed a decrease

(p<0.05) in hematological parameters including HCV, ESR and Hb of

bST-treated dairy animals. The relative difference may be due to stage

of lactation and dose of bST used, because init iation of lactation leads

to an increase in the vascular bed, followed by an increase in blood

volume, whereas Hb concentration is directly related to PCV (Schams

et al. 1975).

57

Table 7

Effect of bovine somatotropic hormone administration on the health of lactating Nili- Ravi buffaloes

Period Diseases Treated

Control

(No. of cases) (No. of cases)

First year

Foot and mouth disease - -

Mastitis 5.0 2.0

Theileriosis - -

Lameness 2.0 4.0

Second year

Foot and mouth disease 11.0 10.0

Mastitis 9.0 7.0

Theileriosis - -

Lameness 1.0 1.0

Third year

Foot and mouth disease - -

Mastitis 4.0 5.0

Theileriosis - 1.0

Lameness - -

Overall

prevalence

Foot and mouth disease 11.0 10.0

Mastitis 18.0 14.0

Theileriosis - 1.0

Lameness 1.0 1.0

58

Table 8

Effect of bovine somatotropic hormone administration on the hematology of lactating Nili –Ravi buffaloes (mean ±SE)

Parameters Study period

Control (0 bST)

Treated (+bST)

Concentration SE Concentration SE

Erythrocyte sedimentation

rate (ESR)

(mm/Ist hr)

Year -1

Year-2

Year-3

Over all

70.50

60.10

80.90

70.50

4.20

6.20

4.10

2.60

71.70

64.30

76.10

70.70

3.80

4.60

5.40

1.54

Hematocrit value

(HCV) (%)

Year -1

Year-2

Year-3

Over all

31.85

36.34

37.12

35.10

0.62

0.46

0.85

0.73

32.27

35.24

36.15

34.55

0.76

0.58

1.10

0.52

Hemoglobin (g/dl)

Year-1

Year-2

Year-3

Over all

12.62

13.63

13.74

13.00

0.19

0.29

0.32

0.30

12.72

13.26

13.80

13.26

0.18

0.24

0.38

0.34

SE = standard error. bST= Recombinant bovine somatotropic hormone mm / Ist hr = millimeter first hour.

Biochemical parameters

The overall (over the 3 years study period) average biochemical

parameters including plasma total protein, total l ipids, BUN,

cholesterol and glucose of the control and bST-treated groups were 8.29

vs. 8.39 mg /dl, 533.0 vs. 531.1 mg /dl, 4.41 vs. 4.11 mmol /L, 162.92

59

vs. 159.2 mg /dl and 55.22 vs. 57.0.3 mg/dl, respectively (Table 9). The

results of all parameters except BUN were non signif icant (P>0.05);

BUN was signif icantly lower (P<0.05) in the-bST treated group

compared to the control. Lower value of BUN is an indication of more

eff icient util ization of protein. Mishra and Shukla, (2004) also found

that bST had no effect on biochemical parameters except BUN of the

dairy Murrah buffaloes. The present results partially agree with those

of Chaiyabutr et al. (2005) who found that bST had no effect on plasma

glucose, total protein and l ipids in dairy cows. In contrast, the present

observations were not in agreement with of Helal and. Lasheen (2008),

who found that the plasma protein and glucose concentrations were

signif icantly increased, while blood urea and cholesterol remained

unaffected in bST-treated (500 mg/14 d) in Egyptian dairy buffaloes.

Mineral metabolites

The overall average serum sodium, potassium and calcium

concentrations of the control and bST-treated groups were 144.90 vs.

137.36, 6.31 vs. 6.6.28 and 4.49 vs. 4.54 meq/L, respectively (Table

10), with none of the differencs being signif icant (P>0.05). McGuffy et

al. (1990) also found sodium chloride and potassium concentrations

were not signif icantly different between bST-treated and control dairy

cows

60

Table 9

Effect of administration bovine somatotropic hormone on blood metabolites in lactating Nili –Ravi buffaloes (mean ±SE)

Parameters Study period

Control (0 bST)

Treated (+bST)

Concentration SE Concentration SE

Protein Mg/dl

Year-1 Year-2 Year-3

Over all

8.48 7.92 8.46 8.29

0.08 0.12 0.20 0.08

8.37 8.11 8.70 8.39

0.07 0.98 0.18 0.07

Lipids mg/dl

Year -1 Year-2 Year-3

Over all

549.9 576.3 472.7 533.0

9.4 21.0 6.6

13.9

564.7 554.1 474.5 531.1

8.5 15.0 3.8

12.7

Blood urea

nitrogen mmol/l

Year -1 Year-2 Year-3

Over all

4.32 a 4.02 a 4.91 a 4.41 a

0.13

0.07

0.16

0.11

3.95 b

3.80 b 4.59 a 4.11 b

0.04

0.05

0.08

0.06

Cholesterol

mg/dl

Year -1 Year-2 Year-3

Over all

173.83 157.03 157.9

162.92

1.8 1.9 5.0

2.44

170.95 154.4 152.2 159.2

2.5 3.4 4.3

2.63

Glucose

G/dl

Year -1 Year-2 Year-3

Over all

55.49 55.37 54.82 55.22

2.1 1.7 2.9

0.86

57.22 56.80 57.0.7 57.03

1.4 2.0 2.4

0.77

SE = standard error mmol = mill i mole bST= Recombinant bovine somatotropic hormone.

61

Table 10

Effect of bovine somatotropic hormone administration on mineral metabolites of lactating Nili –Ravi buffaloes (mean ±SE)

Parameters Study period

Control (0 bST)

Treated (+bST)

Concentration SE Concentration SE

Sodium (meq/l)

Year-1

Year-2

Year-3

Over all

131.70

135.69

155.50

144.90

3.40

4.70

7.20

8.00

124.70

137.79

149.60

137.36

4.50

4.40

8.10

7.10

Potassium (meq/l)

Year -1

Year-2

Year-3

Over all

6.41

6.35

6.17

6.31

0.40

0.34

1.07

0.60

6.31

6.43

6.11

6. 28

0.89

0.41

0.88

0.72

Calcium (meq/l)

Year -1

Year-2

Year-3

Over all

4.71

4.60

4.17

4.49

0.45

0.53

0.70

0.56

4.74

4.63

4.23

4.54

0.43

0.64

0.56

0.55

bST = recombinant bovine somatotropic hormone. SE = standard error meq = mil l i equivalent

62

Economics of milk production

The average daily milk production in bST treated and control

groups was 7.98 and 6.86 kg, respectively throughout the experimental

period. The average price per ki logram of milk in the peri- urban area

was Rs. 25.0. Therefore, the daily net income per animal was Rs 92.00

vs 78.14 in the bST-treated and control groups, respectively. Increased

net income in the bST-treated group was Rs.13.86/d. Hence the net

profit (of bST-treatement) per animal over the 305 d lactation was

Rs.4227.3 (Table 11), thus indicating that the use of bST for increasing

milk production is economical. These results are with agreement with

Tauer and Knoblauch, (1997) who studied the economics of dairy farms

and reported that net farm income increased by $27.0 per cow

compared to the farms where bST was not supplemented to dairy cows.

The economics of using bST may vary depending on farm size and the

management of farm. Kinoshita et al. (2004) who reported that bST had

negative effect in smaller farms as compared to large ones due to

higher cost and lower profit earning abili ty.

63

Table 11

Economics of the use of bovine somatotropic hormone in lactating Nili-Ravi buffaloes.

Parameters Treated (+bST)

Control (0bST)

Av. daily feed intake/ animal (kg)

13.22 12.98

Value of feed @ Rs.7.0 / Kg

92.54 90.86

Cost of bST/animal /day

15.0 -------

Total cost (Feed + bST) /day (Rs.)

107.50 90.86

Av. daily milk yield / animal (kg)

7.98 6.86

Value of milk @ Rs. 25 / kg

199.50 169.00

Net income Rs./day/animal

92.00 78.14

Net daily income in profit (Rs.)

13.86 ------

Net per animal prof it over lactation of 305 Days (Rs.)

4227.30 ----

bST = recombinant bovine somatotropic hormone. Rs. = Pakistan rupees

64

CHAPTER-4

Production performance of lactating Nili-Ravi buffaloes under the influence of somatotropic hormone with varying levels of

dietary energy

ABSTRACT

The study was conducted to determine the effect of dietary energy

on milk yield and its composition under the influence of bST in Nili

Ravi buffaloes. Multiparous buffaloes (n=12) at mid lactation and

similar level of production were selected and randomly divided into

three groups (A, B and C) with four animals in each group in a

completely randomized design. All the experimental animals were

injected bST at fortnightly intervals after 60 ± 3 d postpartum for a

period of 90 d. The nutritional requirements of these animals were met

through a total mixed ration with different densities of energy (low

energy density, 85 %; medium energy density, 100 %; and high energy

density, 115 % of NRC, 2001 standards). The increase in milk yield on

the high energy density (HED) ration was signif icantly higher (P<0.05)

than for the low energy density (LED) and medium energy density

(MED) diets. The difference in the milk yields between the LED and

MED rations was non signif icant (P>0.05). Feed eff iciencies were 0.61,

0.66 and 0.74 on LED, MED and HED diets (P<0.05), respectively.

Nutrient intake (except for ether extract) was signif icantly (P<0.05)

higher on the LED ration than the other two other rations. However,

milk composit ion and body weight gain were similar for all the rations.

Results indicated that 15% higher energy than recommended by NRC,

favored milk production in Nili-Ravi buffaloes when they were injected

with bST hormone.

Key words Bovine somatotropic hormone; Nili-Ravi buffaloes; milk

yield; dietary energy.

65

INTRODUCTION

Bovine somatotropin improves eff iciency of milk yield by

partitioning nutrients toward the mammary gland for increased milk

synthesis (Snatos et al., 1999). Response to bST is dependent on

overall management practices before and after initiation of treatment

(Bauman, 1992). A reduced response might be associated with the

energy and metabolic status of the cow (Leonard and Block, 1997). In

early postpartum, high yielding cows experience a negative energy

balance because of low energy intake relative to energy demand for

maintenance and milk yield, which may l imit their abil ity to respond to

exogenous bST. During periods of negative energy balance endogenous

bST is high, but concentrations of IGF-I and insulin are low and have

ult imately effect on plasma level of l ipitin, which counteract on milk

production of the cow (Block et al. , 2001).

Energy requirements are increased because milk yields are

elevated by bST. The requirements appear to be met primari ly by an

increase in feed intake or, when cows are in negative energy balance,

by lipid mobil ization (Peel and Bauman, 1987). Because the restoration

of body energy stores during late lactation is important to milk yield in

subsequent lactations, dietary fat supplements fed in early lactation and

during bST treatment may help to increase the energy density of the

ration and to improve the metabolic eff iciency of energy uti l ization for

milk production (Srivastava and Mudgal, 1984). However, some studies

have shown that dietary energy concentration in excess of NRC

recommendations does not signif icantly affect responses to bST for any

variables measured (Hemken et al. 1991; Lormore et al. 1990). The

objective of study was to determine the optimum dietary energy level

for lactating buffaloes receiving exogenous bST.

66

MATERIALS AND METHODS

Experimental design

A 90-d feeding tr ial was conducted to determine the optimum

level of energy in the ration of Nil i-Ravi buffaloes being administered

bST. For this study 12 multiparous buffaloes in mid-lactation having

similar milk yield (8.0±1kg/d) and parity were selected from the herd

maintained at the Livestock Experiment Station Rakh Dera Chahal,

Lahore. These animals were randomly divided in to three groups i.e. A,

B and C with four animals in each group. All the experimental animals

were injected with bST (Boostin -250™). The dose level was 250

mg/animal and injection was given at fortnightly intervals during study

period. The study lasted for three months.

Experimental rations

The maintenance and production requirements of individual

animal were met through total mixed rations (TMR) with varying levels

of metabolizable energy (ME). Three isonitrogenous experimental diets

with varying levels of energy (100%, 15% below and 15% above the

recommendations of NRC, 1989) were prepared. The TMR was fed

twice daily to individual animals and water was offered ad libitum. The

ingredients and composition of the rations is presented in Table 1.

67

Table 1

Ingredients and chemical composition of total mixed rations offered to bST treated Nili Ravi buffaloes

Ingredients A B C

LED MED HED

Corn gluten meal 30% 10.00 11.50 19.00

Corn gin - 12.50 15.00

Rice polishing - 10.50 19.00

Wheat bran 25.00 6.00 -

Cotton seed cake 15.00 11.00 -

Sun flower meal 5.50 7.50 6.50

Wheat straw 25.00 24.00 22.00

Molasses 17.50 14.50 13.00

Magalac* - 1.00 4.0

Mineral mix.** 1.00 1.00 1.00

Calcium carbonate 1.00 0.50 0.50

Nutient composition

Dry matter (%) 90.30 90.60 90.80

Crude protein (%) 12.03 12.00 12.07

Metabolizable energy (Mcal/kg) 2.03 2.35 2.69

Neutral detergent fiber (%) 40.88 35.90 33.88

Acid detergent fiber (%) 23.38 20.75 18.70

Calcium (%) 0.84 0.84 0.84

Phosphorus (%) 0.45 0.45 0.45 *Magalac Commercial protected fat *٭ Mineral mixture contained (/kg) Dicalcium phosphate 708g; Magnassium

sulphate 86.4 g; Sodium choloride 189.2 g; Ferous sulphate 8.9 g; Manganese sulphate 4.9 g; Zinc sulphate 2.2 g; Copper sulphate 0.3 g; Potassium iodide 87.7 mg; Cobalt choloride 8.9 mg and Sodium selenate 15 mg.

68

Milk production and composition

Buffaloes were hand milked twice daily at 0300 and 1500 h. Milk

production from the morning and evening was weighed and recorded.

The recording of milk production commenced in the adjustment period

(-1 week) prior to bST treatment and continued for three months

(completion of the trial). Milk samples were collected on weekly basis

and representative samples were used to determine fat %, SNF % by

using Gerber and Rapid methods, respectively described by FAO

(1986). Total solids (TS%) and lactose percentage determined

according to the methods described by AOAC (1990). Crude protein

(CP%) was analyzed with Kjeldahl methods according to BSI (1990).

Dry matter intake

The feed consumption data were recoded daily to evaluate

production eff iciency of the experimental groups. Representative

samples of TMR and orts were analyzed weekly for proximate analysis

according to AOAC, (2001) and the cell wall constituents (NDF% and

ADF%) were analysed according to the procedures described by Van

Soest et al. (1991). The gross energy of the TMR was determined

through the IKA C-2000 Bomb Calorimeter, while metabolizable energy

(M.E) was calculated as 63% of the gross energy (Mandal et al. , 2003).

The analysis was done in Laboratories of Food and Nutrition

Department, University of Veterinary and Animal Sciences, Lahore.

Feed eff iciency of the experimental rations was determined with the

help of following formula

Feed eff iciency = milk produced (kg) / dry matter intake (DMI)

69

The body weights of the experimental animals were regularly recorded

at fortnightly interval using weighbridge available at the farm.

Statistical Analysis

Data obtained on milk production, milk composit ion (fat, SNF,

total solids, milk protein, ash and lactose), DMI, feed eff iciency,

weight gain and nutrient intake were statistically analyzed using

MINITAB software package of statistical analysis following completely

randomized design (CRD). Tukey’s signif icance difference test was

applied to compare the difference between the treatment means. The

statistical model used for all parameters was

Ỵij = µ + tj + єij

Where

µ = Over all means

tj= Effect of treatments ( 1, 2, 3)

єi j = Difference within treatments means

70

RESULTS AND DISCUSSION

Milk production

The average daily milk yields of the animals fed either a low

energy density (LED), medium energy density (MED) or high energy

density (HED) and treated with bST was 7.90, 8.16 and 8.88 kg/d,

respectively (Table 3). The weekly milk production trend of the three

treatment groups is shown in Figure 1. The average daily milk yield of

animals fed HED ration was signif icantly higher (P<0.05) than other

rations but the difference was non signif icant (P>0.05) between MED

and LED. The increase in milk yield of HED group was due to the

effect of their higher energy intake since the dietary protein level and

management was the same for all three groups. Lormore et al. (1990)

and McGuffey et al. (1990) reported that administration of bST causes

a shift in partioning of available energy towards milk yield at the

expense of body tissue, so that an energy dense diet may be needed.

Contrary to the present f indings Hemken et al. (1991) reported that

feeding bST-treated dairy cows a diet higher in energy than NRC

recommendation had no signif icant impact on milk yield. Also, contrary

to the f indings of the present study Tarazon-Herrera et al. (2000) found

that milk yield response was greater for bST-treated cows fed a low

rather than a high-energy diet. Difference may be attributed to stage of

lactation.

71

0.00

2.00

4.00

6.00

8.00

10.00

12.00

1 2 3 4 5 6 7 8 9 10 11 12

Milk

yie

ld (

kg/d

)

Weeks

LED

MED

HED

Figure 1 Trend of milk production in animals given low, medium and

high-energy diets.

Milk composition

The average milk fat, SNF, total solids, protein and lactose percent are

shown in Table 3. The energy density of the diet had no effect (P>0.05) on

any of these parameters. Bauman, (1989) found that bST treatment did not

affect milk composit ion due to the homeorhetic control involved in the

metabolism of lipid, carbohydrates and amino acids. The results of present

study are also consistent with previous f indings of Terrazon-Herraera et al.

(2000) who did not record any effect of bST administration on milk

composition (SNF, fat, protein and lactose) in western dairy cows with

varying level of dietary energy.

72

Table 3.

Effect of bovine somatotropic hormone administration on productive performance in lactating Nil i- Ravi buffaloes fed varying levels of dietary energy (mean± SE)

Variables LED (+ bST)

MED (+ bST)

HED (+ bST)

Milk production (kg/d) 7.90 ± 0.11 a 8.16 ± 0.06ab 8.88 ± 0.0.20 c

Milk composit ion (%) Fat

SNF

TS

CP

Lactose

7.00 ± 0.12 9.02 ± 0.12 16.07 ± 0.03 3.35 ± 0.05 4.65 ± 0.08

6.46 ± 0.16 8.94 ± 0.09 15.40 ± 0.09 3.27 ± 0. 10 4.63± 0.08

7.09 ± 0.13 8.95 ± 0.09 16.05 ± 0.08 3.24 ± 0.07 4.86 ± 0.09

Body weight gain (kg) 5.75 ± 0.64 4.75 ± 1.10 5.00 ± 1.08

Feed eff iciency (kg of milk yield/ DMI)

0.61 ± 0.01a 0.66 ± 0.01b 0.74± 0.01c

Means with different superscripts in a row differ significantly (P< 0.05) bST = bovine somatotropic hormone Fat= crude fat SNF = solid not fat TS = total solid CP = crude protein

Body weight change

The average weight gain over the tr ial period on ration LED,

MED and HED was 5.75, 4.75 and 5.00 kg, respectively (Table 3) and

these did not differ signif icantly (P>0.05). Srinivasa-Rao and

Rangandham (2000) and Jorge et al. (2002) found that body weights

were not affected on administering bST to buffaloes, whilst Huber et al.

73

(1997) found that body weights were increased up to 37% in cows

injected with bST for four consecutive lactations. However, Moallem et

al.(2001) observed a decrease in body weights in dairy cows despite an

increased DMI after bST treatment.

Feed intake and feed efficiency

There was a signif icant decrease (P<0.05) in DMI as the energy

density of the diet increased (Table 4). However, there was a

concomitant improvement in average feed eff iciencies (milk yield/kg

DMI); 0.61, 0.66 and 0.74 for LED, MED and HED, respectively (Table

3 and Figure 2). The improvement in feed eff iciency with energy

density of the diet was l inear; with an improvement (P<0.05) of 8.2 and

21.3% for MED and HED groups, respectively compared to LED.

Chill iard (1989) suggested that the magnitude of increase in feed intake

depends upon milk yield and energy density of diet. Results of this

study indicate that feed intake increases as energy density of the diet

decreases Lormore et al. (1990) and Drackley et al. (2003) also found

feed eff iciency was improved in bST-treated dairy cows fed a high-

density ration. This was achieved by altering the nutrient in a manner

that provides the mammary glands increased access to substrate

supplies that may be uti l ized for milk production rather than increasing

adipose tissue store. Bauman et al. (1992) and Chalupa et al. (1989)

observed that daily nutrient requirements were increased by an amount

equal to the increase in milk, and productive eff iciency (milk per unit

of feed) was improved because a greater proportion of the nutrient

intake was used for milk synthesis.

74

0.610.66

0.74

0.00

0.10

0.20

0.30

0.40

0.50

0.60

0.70

0.80M

ilk y

ield

/ D

MI

LED MED HED

Dietary groups

Figure 2. Average feed eff iciency of bST-treated Nil i-Ravi buffaloes

fed low, medium and high energy density rations

Nutrients intake

The average daily nutrient intakes (crude protein, NDF, ADF,

ether extract and Ash) are shown in Table 4. As the energy density of

the diet decreased, individual nutrient intakes (with the exception of

ether extract fraction) increased (P<0.05). This was to overcome the

energy requirements in response to increasing milk yield with bST

because the magnitude of increase in DMI depends upon the response in

milk yield, the method of bST administration and the energy density of

the diet (Chil l iard, 1989). Bauman et al. (1985), Moallem et al. (2000)

and Helal and Lasheen, (2008) concluded that DMI increased linearly

as milk production increased when animals are injected with bST. This

increase in nutrients may be attributed to the homeorhetic regulation of

bST. Results of the present study however, indicate that responses in

75

DMI will be modif ied by the energy density of the diet being fed to the

animals being administered bST.

Table 4.

Effect of bovine somatotropic hormone administration on daily nutrient intake of in Nil i Ravi buffaloes fed varying level of dietary energy (mean ± SE)

Variables LED (+ bST)

MED (+ bST)

HED (+ bST)

Dry matter intake (kg/d) 11.67 ± 0.06 a 11.17 ± 0.11b 10.85 ± 0.06c

Crude protein (kg/d) 1.40 ± 0.01a 1.34± 0.01b 1.30± 0.04 c

Neutral detergent f iber

(kg/d) 4.77 ± 0.02a 4.00 ± 0.04b 3.73 ± 0.06

Acid detergent f iber

(kg/d) 2.70 ± 0.01a 2.30 ± 0.02 b 2.02 ± 0.01c

Ether extract (kg/d) 0.37 ± 0.06a 0.39 ± 0.01 b 0.46± 0.01 c

Ash (kg/d) 1.56 ± 0.01a 1.55 ± 0.01b 1.27 ± 0.00c

Means within a row bearing different superscripts differ signif icantly (P<0.05) DM = dry matter CP = crude protein, NDF= neutral detergent f iber ADF = acid detergent fiber EE = ether extract.

76

CHAPTER-5 Publication 1

Effect of bovine somatotropic hormone on the

productive performance of Nili-Ravi buffaloes

M.A. Jabbar1, I. Ahmad1, S. Javid2, M.A. Chaudhry2, R.H. Usmani3 1 Department of Animal Nutrition, University of Veterinary and Animal Sciences,

Lahore, Pakistan 2 Livestock Production Research Institute, Bahadurnagar, Okara

3 Pakistan Agriculture Research Council, Islamabad Corresponding author M.A. Jabbar. Department of Animal Nutrition University of Veterinary and Animal Sciences, Lahore, Pakistan - Tel. +92-42-7236374 - Email [email protected]

ABSTRACT The study was conducted on 21 Nili-Ravi lactating buffaloes with similar milk production and stage of lactation. They were randomly divided into three groups A, B and C with 7 animals in each group. The group A was injected with full dose of bST hormone (250 mg/animal) with trade name of Boostin-250 at an interval of 14 days, while animals in group B were given injection on alternate days with divided dose of 36 mg/animal. The group C was kept as control. The duration of study was 5 months and the animals were kept on green fodder supplemented with concentrate ration @ half of milk production. The concentrate ration had 17.2% CP and 72.0% TDN. The milk production was 8.06 ± 1.49, 7.85 ± 1.08 and 6.81 ± 1.80 liter in respective groups. The milk production increased by 18.35% and 15.27% in group A and B compared with group C (control). However, the increase was statistically non-significant (P>0.05). The difference in Fat, SNF, and TS percent was nonsignificant among the groups. The service period and services per conception were improved in both the treated groups. The body weight in sub-group A, B and C were similar and none of them lost weight over the study period. The difference among hematological and biochemical parameters was also non-significant.

Key words Bovine Somatotropic hormone, Nili-Ravi buffaloes, Milk production, Reproduction.

INTRODUCTION

With the increase in demand of foods for the ever-growing human population, there is a need to increase the livestock productivity. Feed is a major cost in the Production, which accounts for 60-65% of the total production cost. Hence new technologies are required to attain greater feed efficiency. Bovine somatotropic hormone is one of the first potential biotechnology products in animal production, which improves productive efficiency (Bauman, 1992). Ital.J.anIm.Sci. 6, (Suppl. 2), 1039-1042, 2007 1039

77

VIII World Buffalo congress

In dairy cattle use of bST resulted in 12-25% improvement in milk yield and 5 to 15% feed efficiency with out any substantial alteration in the composition of milk (Burton et al., 1994). Bovine somatotropin reduces the proportion of maintenance requirement out of total nutrient intake (Bauman et al., 1989). Buffalo is the main dairy animal in Pakistan. In spite of large number and good adaptabilility, per animal is very low as compare to Western dairy cattle breeds. Usmani et al. (1991) reported that increased milk production in buffaloes with (bST) is cost effective and harmless and is good contribution towards the milk production. Most of available information’s on buffalo are based on short-term studies. Present study was planned to determine the effect of long-term use of bST on productive, reproductive, physiological / biochemical parameters in Nili-Ravi buffaloes.

MATERIAL AND METHOD S

For the study twenty-one multiparous lactating Nili-Ravi buffaloes with similar stage of lactation and production were selected and kept at Livestock Experiment Station Bahadurnagar, Okara. They were randomly divided into three groups A, B and C. The group A was treated with (bST) hormone with trade name of Boostin-250 manufactured by LG Life Sciences, Ltd. South Korea, at 14 days interval with full dose of 250 mg. Group B was injected at alternate day at divided dose of 36mg/day whereas group C was control (without bST). The research trial continued for five months. The experimental animals were fed green fodders and concentrates. The maintenance requirement of the animals was met by available green fodder of 10% of body weight while production requirements were met through concentrate ration with 17.1% CP and 72.15 TDN. The following parameters were studied 1 Milk production and composition Milk production of individual animals of both groups was recorded daily at the morning and evening milking while milk samples was collected individually on weekly basis. Milk samples were analyzed for Fat, Solids Not Fat (SNF) and Total Solids (TS) percentage through method described by Aggarwala and Sharma, (1961). 2. Feed consumption and body weight Body weights were recorded on monthly basis. Weighed quantities of concentrate ration and fodder were offered to animals. Feed and fodder samples were collected on weekly basis and analyzed for proximate analysis (AOAC, 1995). 3. Reproductive Parameters For reproductive performance data on postpartum estrous, length of estrous period/ (days open) and no of services per conception were recorded. 4. Physiological and Biochemical parameters Blood samples were collected on fortnightly basis and were analyzed for the hematological and biochemical parameters given in Table 1. The data obtained on various parameters were subjected to statistical analysis by using complete randomized design. Duncan’s multiple range test was applied to compare the difference between the means (Steel et al., 1997).

1040 Ital.j.anim.Sci.vol. 6, (Suppl. 2), 1039-1042, 200

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VIII World Buffalo congress

RESULTS AND CONCLUSIONS

Milk production The average milk production of groups A, B and C were 8.06 ± 1.49, 7.85 ± 1.08 and 6.81 ± 1.80 liters, respectively. The increase in milk production was 18.35 and 15.27% compared with group C (control) but statistical analysis revealed that the difference was non-significant. The results of the present study are in agreement with the findings of Jorge et al., (2002), Mishra and Shukla (2004) who found that the increase was 14.93 to 32.4% in treated Murrah and Kundi buffaloes but increase depended on dose rate of bST. Milk composition The average Fat, SNF and TS percentage of the group A, B and C was 6.62 ± 0.21, 6.67 ± 0.07, and 6.54 ± 0.07; 9.25 ± 0.28, 9.33 ± 0.13 and 9.11 ± 0.11; 15.90 ± 0.48, 16.04 ± 0.13 and 16.10 ± 0.85 percent, respectively. Slight increase was observed in milk contents of treated animals over the control but statistical analysis revealed that there was no significant difference. The results of study were similar to the results of Jorge et al., (2002) and Mishra and Shukla (2004) who reported that bovine samototropin did not influence milk composition of dairy cows, Murrah and Kundi buffaloes. The results differed from the findings of Huber et al., (1997) who reported that fat percent decreased in dairy cows with long term use of bST hormone. Body weights The average body weights in groups A, B and C were 537.19 ± 50.89, 550.62 ± 89.43 and 528.30± 30.80 kg, respectively. There was non-significant difference in body weights of bST treated groups than control. The results of the study were similar to the results of Srinivasa-Rao and Ranganadham (2000) who found that body weights were not affected by supplementation of bovine somtotropin. Results of present study differed from Huber et al., (1997) who found that body weights were enhanced up to 37% in cows injected with bST for four conescutive lactations. Reproductive performance The service period was 71.4, 67.33 and 112.36 day while services per conception were 1.2, 1.16 and 1.58 in group A, B and C, respectively. Apparently it was found that there was improvement in reproductive parameters in buffaloes receiving bST. The results of the study are supported by the findings of Dahoo et al., (2003) who found no adverse effect of bST on reproductive performance in dairy cows rather some measures tended to improve with bST. Blood metabolites The means haematological and biochemical values of the control and treated animals are shown in Table 1. All such parameters did not vary (P > 0.05) among experimental groups. The results of study are similar to the results of Kweon (2000) who concluded that bST had no effect on hematological and biochemical values of the dairy animals.

Ital.j.anim.Sci.vol. 6, (Suppl. 2), 1039-1042, 2007 1041

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VIII World Buffalo congress

Table 1. Mean (± S.D.) values of different haematological and

biochemical para-meters. Parameters Group –A Group-B Group-C

ESR mm (Ist h) 64.15 ± 7.93 55.15 ± 29.03 62.59 ± 19.05

Haemetocrit value% 32.0 ± 3.59 32.37 ± 2.30 31.96 ±1.70

Glucose mg /dl 56.36 ± 3.05 55.49 ± 4.62 57.27±2.32

Haemoglobin mg /dl 11.43 ± 0.62 11.85 ± 0.34 12.04 ± 0.53

Total protein mg /dl 7.67 ± 0.43 7.96 ± 0.38 7.49 ± 0.36

Cholestrol mg /dl 173.90 ± 34.9 175.90 ± 17.4 185.12 ±14.1

Urea mg /dl 3.79 ± 0.36 3.85 ± 0.12 3.92 ± 0.29

Calcium mg /dl 9.54 ± 1.32 9.23 ± 1.30 9.63 ± 1.16

Sodium meq / liter 177.31 ± 30.1 165.29 ± 28.2 177.39 ± 30.2

Potassium meq / liter 36.48± 4.52 34.22 ± 6.85 35.81 ± 5.70

REFERENCES - Aggarwala, A.C. and Sharma, R.H. 1961. A laboratory manual of

milk inspection. 4th ed. Asia Publishing House London. AOAC. 1995. Offcial

Methods of Analysis. 15th ed. Association of Analytical Chemists, Arlington,

Virginia, USA. Bauman, D.E. (1992) Bovine somatotropin review of an emerging

animal technology. J. Dairy Sci.75 3432-3451. Bauman, D. E., Hart, D.L. Crooker,

B.A., Partridge, M.S., Garrick, K. D., Sandles, K., Erb, H.N., Franson, S.E., Hrtnell,

G.F.and Hintz, R.L 1989. Long-term evaluation of prolonged-release formulation of

N-methionyl bovine somatotropin in lactating dairy cows. J. Dairy Sci. 72 642.

Burton, J.L., Block, E., Glimm. R., 1994. A review of bovine growth hormone. Can.

J. Anim. Sci.74 167-201. Dohoo, I.R, DesCoteaux, L., Leslie, K., Fredeen, A.,

Shewfelt, W. and Preston, A., 2003. A meta analysis review of the effects of recombinant

bovine somatotropin. 2. Effects on animal health, reproductive performance, and culling.

Can. J. Vet. Res. 67 252-64. Huber, J. T., Fontes, JR. Z. WU. C., Sullivan, J. L., Hoffman,

R. G. and Hartnell, G. F., 1997. Administration of recombinant bovine somatotropin to dairy

cows for four consecutive lactations, J Dairy Sci 802355-2360. Jorge, M.A., Gomes,

M.I.F.V. and Halt, R.C., 2002.

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VIII World Buffalo congress

Effect of recombinant bovine somatotropin (bST) utilization on milk production from

buffaloes. R. Bras Zootec. 31 213-222. Kweon, U.G.K.T., Nam, H.S., Kim, S.K., Yun, J. S.

and. Hang, B. J., 2000. Effect of recombinant bovine somatotrpic hormone injection time on

haemotological values and blood metabolic substrates in Holstein steers. Kor .J. Anim. Sci.

42 269-278. Mishra, A. and Shukla, D. C., 2004. Effect of Recombinant Bovine

Somatotropin (Boostin-250) on blood metabolites and milk yield of lactating buffaloes.

Asian-Aust. J. Anim. Sci. 17 1232-1235. Srinivasa-Rao, K. and Ranganadham, M., 2000.

Effect of bovine somatotropin on milk production and composition in lactating Murrah

buffaloes. Indian J. Dairy Sci. 53 46-50. Steel, R.G.D., Torrie, J.H and Dickey,. D.A., 1997.

Principles and procedures of statistics. A biochemical approach (3rd ed.) McGraw Hill Book

Co. Inc., New York, USA. Usmani, R.U., Khan, A.G. and Author, I.H., 1991. A review of

growth hormone biotechnology for increased milk production. Pakistan.Vet.J.11 100.

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CHAPTER-5 Publication 2

Short communication

PERFORMANCE OF LACTATING BUFFALOES ADMINISTERED BOVINE SOMATOTROPIN HORMONE

I. Ahmad, M. A. Jabbar1, T. N. Pasha and M. Abdullah2 Department of Animal Nutrition

University of Veterinary and Animal Sciences, Lahore-5400, Pakistan (Received 12 July, 2006)

ABSTRACT Nili-Ravi buffaloes (14; 476.9t57.2 kg BW; 8.88t0.98 1 milk/d; 90t6.5 days in milk) randomly divided into two equal groups. Animals in both the groups were offered conventional TMR, while animals in 2nd group were injected bovine somatotropin (bST) hormone (250 mg/animal) at an interval of 14 days for 60 days. The milk production was increased by 7.0% in bST group over that of control group and the increase was statistically non-significant. However, the feed efficiency for milk production was improved (P< 0.05) in treated group. The milk composition, body weight gain and digestibility of nutrients were similar in both groups. It was concluded that there was no effect of administering bST hormone @ 250mg/animal on milk yield and its composition in buffaloes. (Indian J. Anim. Nutr. 2007, 24(1) 5254)

Key words Bovine somatotropin, Milk production, Feed efficiency, Buffaloes

Bovine somatotropin improves the

productive efficiency specially milk yield per unit of feed input. In dairy cattle, use of bST resulted in 12-25% improvement in milk yield and 5 to 15% feed efficiency without any substantial alteration in the composition of milk (Ludri et al., 1989; Rose et al., 2004). The higher productive response was attributed to increased synthesis of lactose, fat and pro-tein in mammary glands (Bauman et al., 1988). There is not much information available about use of bST hormone in buffaloes. The present experiment was conducted to study effect of bST on feed efficiency in lactating buffaloes.

Nili-Ravi buffaloes (14; 476.9 to 57.2 kg BW; 8.88to.98 kg/d milk yield and 90t6.5 days in milk) randomly divided in to two equal groups. Animals in both the groups were offered conventional TMR, while animals in 2nd group were injected recombinant bovine somatotropic hormone (bST) @ 250 mg/animal (Boostin -250) at fortnightly interval for 60 days.

Total mixed ration (maize grain 9.0, rice polish 8.0, wheat bran 7.0, cotton seed cake 10.0, cotton seed meal 2.0, maize gluten (30%) 13.0, rape seed meal 7.0, rice straw 30.0, molasses 12.0, mineral mixture 2.0%) containing 12.5% CP and 65.0% TDN (NRC, 2001) was offered.

Table 1. Intake and digestibility of nutrients in buffaloes administered bovine somatotropic hormone

Parameters TMR TMR-bST

Pooled SE

DM intake % BW 2.3 2.4 0.9 Digestibility of nutrients, %

DM 67.3 71.6 0.9 CP 65.0 66.3 1.5 CF 76.7 77.4 1.0 EE 75.5 75.9 1.5 Ash 71.3 71.1 0.7

1Corresponding author E-mail: [email protected]; 2Department of Livestock Production

82

Effect of bovine somatotropin on the performance of buffaloes

. The experimental animals in both the groups were fed available green fodder @5 kg/animal/d to fulfill the requirement of vitamin A. Milk yield was recorded daily while milk samples were analyzed for Fat, SNF and TS weekly (Aggarwala and Sharma, 1961). Feed samples were analyzed for proximate principles (AOAC, 1990), and feed consumption data were recorded. During the last week of study a digestibility trial on 3 animals from each group was conducted. Representative feed and fecal samples were collected and analysed. Data was statistically analyzed by using t-test (Steel et al., 1997). There was no difference in the digestibility of nutrients in the two groups (Table 1), confirming the earlier report in cows (Santos et al., 1999). However, Newbold et al., (1997) reported that digestibility of nutrients was increased in bST treated animals.

The daily milk yield of groups A and B was 8.25 and 8.80 kg respectively (Table 2), Milk production increased by 7% in group B over that of group A though the difference was non significant. Contrary to the finding of the present study, increased milk production has been reported in many studies on administration of bST hormone. Jorge et al., (2002) observed 32.8% increase with 4% FCM yield in Murrah buffaloes when injected with 500 mg of bST/animal after 14 day interval. Relatively higher increase could be due to higher dosage level used in studies. The average milk fat, SNF, TS and protein percent of both the groups were similar (Table 2). The results of study were similar to those of Mishra and Shukla, (2004) who also did not record any effect of bST administration on milk composition in western dairy cows and Kundi buffaloes.

There was no difference in body weight gain of both groups. The results of the study were similar to the results of Jorge et al., (2002), but Huber et al., (1997) reported that body weights were increased up to 37% in cows injected with bST for four consecutive lactations.

Table 2. Performance of buffaloes on bST administration.

Parameters TMR TMR-bST

Pooled SE

Milk yield, kg/d 8.3 8.8 0.50 Milk composition,

%

Fat 6.7 6.9 0.04 SNF 9.1 9.2 0.10 TS 15.7 16.1 0.20 Protein 3.6 3.7 0.04 Cain in BW, kg 12.9 14.3 14.30 FC, kg/d 12.2 12.4 0.04 PCE 1.5a 1.4b 0.01

Figures with different superscripts in a row differ sig-nificantly, P<O.05; FC - Feed Consumed; FCE-Feed conversion efficiency, kg/kg milk yield

The feed efficiency was significantly improved (P<0.05) in bST treated group confirming the earlier report (Annexstad et al., 1990). It was con-cluded that there was no effect of administration of bST @250mg /animal on milk yield and its composition in Nili-Ravi buffaloes.

REFERENCES

Aggarwala, A. C. and Sharma, R. H. (1961) A laboratory manual of milk inspection. 4th ed. Asia publishing house London.

Annexstad, R. J., Oiterby, D. E., Linn, J. G, Hanse, W. P., Soderholm, C. G and Wheatstone, J. E. (1990) Somatotropin treatment for consecutive lactations. J. Dairy Sci.. 73 2423-2436.

AOAC. (1990) Offcial Methods of Analysis. 15th ed. Association of Analytical Chemists, Arlington, Virginia, USA

Bauman, D. E., Peel, C. J., Steinhour, W. D., Reynolds, P. 1., Tyrell, H. E, Brown, A. C. G and Haland G. L. (1988) Effect of bovine somatotropin on metabolism lactating dairy cows Influence on rates of irreversible loss and oxidation of glucose and non-esterified fatty ac-ids. J. Nutr., 18 1031-1040.

Huber, J. T., W. U., Z., C. Fontes, J. R., Sullivan, L., Boffman, R O. and Hartnell, O. E. (1997) Administration of recombinant bovine somatotropin to dairy cows in four consecutive lac-tations. J. Dairy Sci., 80 2355-2360.

Jorge M. A., Gomes M. I. E. V. and Halt, Re. (2002). Effect of recombinant bovine somatotropin (bST) utilization on milk production from buffaloes. R. Bras Zootec., 31 213-222.

Ludri, R. S., Upadhyay, R. C., Singh, M., Guneratne, J. R. and Basson R. P. (1989). Milk production in lactating buffaloes receiving recombinantly produced bovine somatotropin. J. Dairy Sci., 72 2283-2287.

83

Newbold, J., Heap, A. R. B., Prosser, E. G, Pipps, R. H., Adrianes, E. and Hard. D. L. (1997). The effect of bovine somatotropin and diet on somatotropin binding sites in hepatictissue of lactating dairy cows. J. Dairy Sci., 80 10851091.

NRC (2001) Nutrient Requirements of Dairy Cattle. 7th revised edition, National Academy Press, National Research Council. Washington, DC. pp.147.

Rose, M. T., Weekes, T. E. C. and Rowlinson, P. (2004) Individual variation in the milk yield response to bovine somatotropin in dairy cows. J. Dairy Sci., 87 2024-2031.

Mishra, A. and Shukla, D.C. (2004) Effect of recombinant bovine somatotropin (Boostin-250) on blood metabolites and milk yield of lactating buffaloes. Asian-Aust. J. Anim. Sci., 17 1232-1235.

Santos, J. E. P., Huber, J. T., Theurer, C. B., Nussio, L. G., Nussio, C. B., Tarazon, M. and LimaFilho, R. O. (1999) Performance and nutrient digestibility by dairy cows treated with bovine somatotropin and fed diets with steam-flaked sorghum or steam-rolled com during early lactation. J. Dairy Sci., 82 404-411.

Steel, R. G. D., J. H. Torrie and D. A. Dickey (1997) Principles and procedures of statistics. A Biochemical Approach (3nded.) McGraw Hill Book Co.Inc., New York, USA

84

Chapter-6

SUMMARY

Use of bovine somatotropic hormone (bST) for increased milk

production has been widely investigated in dairy cattle, whereas very

lit t le work has been done in buffaloes. To observe the effect of bST on

buffalo for long term duration study was planned with the objectives to

investigate the effects of long term use of bST on milk production, milk

composition, reproduction, hematological and biochemical parameters

in Nili-Ravi buffaloes. For this study 30 lactating Nili-Ravi buffaloes

with similar milk production and stage of lactation were selected and

randomly divided in to two groups A and B with 15 animals in each

group. The group A (0 bST) served as control while animals in group B

(+bST) were given injection of bST (250 mg Boostin-250/animal) at

14d interval. Nutritional requirements of experimental animals were

met through available green fodder (45-50kg/day) supplemented with

concentrate ration @ half of milk production. The milk production was

signif icantly (P<0.05) increased by 18.04 % in treated group compared

with control. The results showed that there was no signif icantly

variations in parameters like milk composit ion, dry period and lactation

length, calving interval in both the groups. The postpartum estrous

period and service period were signif icantly (P< 0.05) improved which

reflected positive effect of bST on reproductive parameters. However,

the difference in services per conception was non-signif icant. Small

variations were found in the prevalence of contagious and non

contagious diseases in both experimental groups during the study

period. The differences among body weights, hematological and

biochemical parameters were also non-signif icant expect blood urea

85

nitrogen (p< 0.05). The proceeds over a lactation period of 305 days

was PKR. 4227.0 with the use of bST.

Second tr ial was conducted to study the effect of dose interval of

bST in Nili-Ravi buffaloes. For the proposed study 21 Nili-Ravi

lactating buffaloes with similar milk production and stage of lactation

were randomly divided into three groups A, B and C with 7 animals in

each group. The group A was injected with full dose of bST hormone

(250 mg/animal) with trade name of Boostin-250 at an interval of 14

days, while animals in group B were given injection on alternate days

with divided dose of 36 mg/animal. Group C was kept as control.

Duration of study was 5 months and the animals were kept on green

fodder supplemented with concentrate ration @ half of milk production.

The concentrate ration had 17.2% CP and 72.0% TDN. The milk

production increased by 18.35% and 15.27% in-group A and B

compared with group C (control) but increase was non-signif icant

(P>0.05) . Similarly data revealed that dose interval had no affect on

milk contents, reproductive and hematological parameters in all the

experimental groups.

In a third trial feed digestibi l i ty and eff iciency for milk

production was studied. For the study fourteen Nili-Ravi buffaloes at

their mid lactation with almost same level of milk production were

randomly divided into two groups A and B with seven animals in each

group. The group A was kept as control, while group B was injected

bST hormone (250 mg/animal) at an interval of 14 days and continued

for 60 days. The nutritional requirements of animals in both the groups

were met through TMR according to NRC recommendations. The milk

production was increased by 7.0% in treated group (B) as compared

with control group (A) and the increase was statistically non-signif icant

86

(P>0.05). However, the feed eff iciency for milk production was

signif icantly improved (P< 0.05) in treated group. The differences in

milk composition (Fat, SNF, TS and Protein percent) body weight gain

digestibi l ity of dry matter and other nutrients in treated and control

groups were found non-signif icant (P>0.05).

The fourth trial was conducted to determine the effect of energy

on milk production and its quality under the influence of bST hormone

in Nili-Ravi buffaloes. Multiparous (n=12) buffaloes with mid lactation

and similar level of milk yield were selected and randomly divided in

to three groups i.e. A, B and C with four animals in each group. All the

experimental animals were injected bST with trade name of Boostin -

250. The dose level was 250 mg per animal and injection was given at

fortnightly interval during study period. The nutritional requirements

of three groups animals were met through TMR with varying levels of

energy (15% low and 15% above the recommendations of NRC). The

milk yield was signif icantly higher (p<0.05) on medium and high

energy ration but the difference of milk yield was non signif icant

(p>0.05) between medium and low energy diets. The milk components

and body weight gain were similar on all rations, while feed eff iciency

and nutrient intake (except ether extract) in low energy diet was

signif icantly higher (p<0.05) from two other rations. It may be

concluded that 15% higher energy than recommended by NRC favoured

milk production in Nil i Ravi buffaloes when they were injected bST

hormone.

Conclusion

On the over all there was consistency of results for milk

production and milk composition with reference to available l iterature.

However, some reproductive parameters including postpartum estrus

87

and service period were signif icantly improved with the use of bST

hormone. This effect has not been reported in the previous li terature

which needs to be further investigated and verif ied. Similarly the dose

level in buffaloes needs to be further studied.

88

LITERATURE CITED

AboEl-Nor, S. A. H., A. M. Mona, Abd. El-Gawad, and M.H. Abd El-

Salam. 2007. Effect of somatotropin and monensin on the

yield and quality of milk and tal laga cheese from Egyptian

buffaloes. Intl. J. Dairy Sci. 2 58.

Agarwal, S. K., and O. S. Tomar. 1998. Reproductive Technologies in

Buffaloes. Indian Veterinary Research Institute, Izatnagar,

India.

Aggarwala, A.C. and R.H. Sharma. 1961. A Laboratory Manual of Milk

Inspection. (4th ed.) Asia Publishing House London, UK.

Annexstad, R.J., D.E. Oiterby, J.G. Linn, W.P. Hanse, C.G. Soderholm,

and J.E. Wheatstone. 1990. Somatotropin treatment for

consecutive lactations. J. Dairy Sci. . 73 2423.

AOAC. 1990. Solids (Total) in milk Official Methods of Analysis

No.925.21. Association of Official Analytical Chemists

Inc., Arlington, Virginia, USA.

AOAC. 1995. Official Methods of Analysis. 15th ed. Association of

Analytical Chemists, Arlington, Virginia, USA

Asdell, S.A. 1932. The effect of the injection of hypophysial extract in

advanced lactation. Anim. J. Phys. 100 137.

Asimov, G.J., and N.K. Krouze. 1937. The lactogenic preparations from

the anterior pituitary and the increase of milk yield in cows.

J. Dairy Sci. 20 289

89

Austin, C.L., J.D. Schingeothe, and D.P. Casper. 1991. Influence of

bovine somatotropin and nutrition on production and

composition of milk from dairy cows. J. Dairy Sci. 74 3920.

Baer, R.J., K.M. Tieszen, D.J. Schingoethe, R.D. Shaver and R.M.

Cleale. 1989. Composit ion and flavor of milk produced by

cows injected with recombinant bovine somatotropin. J.

Dairy Sci. 72 1424.

Baldi, A. 1999. Manipulation of milk production and quality by use of

somatropin in dairy ruminants other than cows. Domest.

Anim. End. 17 131.

Baldwin, R.L., and J.R. Knapp. 1993. Recombinant bovine

somatotropin's effects on patterns of nutrient uti lization in

lactating dairy cows. Anim. J. Clin. Nutr. 58(Suppl.1) 282.

Barbano, D.J., J.M. Lynch, D.E. Bauman, G.F. Hartnell, R.L. Hintz,

and M.A. Nemeth. 1992. Effect of a prolonged-release

formulation of N-methionyl bovine somatotropin

(sometribove) on milk composition. J. Dairy Sci. 75 1775.

Bauman, D.E., and W.B. Currie. 1980. Partioning of nutrients during

pregnancy and lactation a review of mechanisms involving

homeostasis homeorhesis. J. Dairy Sci. 63 1514.

Bauman, D. E., M. J. DeGeeter, C. J. Peel. G. M. Lanza, R. C. Gorewit,

and R.W. Hammond. 1982. Effect of recombinantly derived

bovine growth hormone on lactational performance of high

yielding dairy cows. J. Dairy Sci. 5(Suppl.1) 121.

90

Bauman, D.E., P.J. Eppard, M.G. DeGeeter, and G.M. Lanza. 1985.

Responses of high producing dairy cows to long term

treatment with pituitary Bovine somatotropic hormone and

recombinant Bovine somatotropic hormone. J. Dairy Sci. 68

1352.

Bauman, D.E., C.J. Peel, W.D. Steinhour, P.J. Reynolds, H.F. Tyrrell,

A.C.G. Brown, and G.L. Haland. 1988. Effect of bovine

somatotropic hormone on metabolism of lactating dairy

cows influence on rates of irreversible loss and oxidation of

glucose and nonesterif ied fatty acids. J. Nutr. 118 1031.

Bauman, D.E., D.L. Hard, B.A. Crooker, M.S. Partridge, K. Garrick,

L.D. Sandles, H.N. Erb, S.E. Franson , G.F. Hartnell, and

R.L. Hintz. 1989. Long-term evaluation of a prolonged-

release formulation of N-methionyl bovine somatotropin in

lactating dairy cows. J. Dairy Sci. 72 642.

Bauman, D.E. 1992. Bovine somatotropic hormone. Review of an

emerging animal technology. J. Dairy Sci. 75 3432.

Bauman, D.E., and R.G. Vernon. 1993. Effects of exogenous bovine

somatotropic hormone on lactation. Annu. Rev. Nutr. 13

437.

Bauman, D.E., R.W. Evertt, W.H. Weiland, and R.J. Collier. 1999.

Production responses to bovine Bovine somatotropic

hormone in northeast dairy herds. J. Dairy Sci. 82 2564.

Benjamin, M. M. 1985. Outline of Veterinary Clinical Pathology. 3rd

ed. Iowa State University Press, Iowa, USA. Pp 64.

91

Bil ly, T.R., A. Sozzi, M.M. Lopes, F.T. Sillvesre, A.D. Ealy, C.R.

Staples, and W.W. Thacher. 2006. Pregnancy, bovine

somatotropin, and dietary n-3 fatty acids in lactating dairy

cows. Ovarian, conceptus, and growth hormone-insulin-like

growth factor system responses. J. Dairy Sci. 89 3360.

Block, S.S., W.R. Butler, R. A. Ehrhardt, A.W. Bell, M.E. Van

Amburgh, and Y.R. Boisclair. (2001). Decreased

concentration of plasma leptin in periparturient dairy cows

is caused by negative energy balance. J. Endocrinol. 17 339.

Bremmer, D.R.; T.R. Overton, and J.H. Clark. 1996. Production and

composition of milk from Jersey cows administered bovine

somatotropin and fed ruminally protected amino acids. J.

Dairy Sci. 80 1374.

Boynd R.D., and D.E. Bauman. 1989. “Mechanisms of action for

somatotropin in growth”. P 257. in D.R. Campion, G. J.

Hausman, R. J. Martin (eds.). Animal Growth Regulation.

New York, Plemun press.

BSI. 1990. Determination of the nitrogen content of l iquid milk. In

Methods for chemical analysis of l iquid milk and cream.

BSI1741, Brit ish Standard Instituation, London, UK.

Burton, J.L., B.W. McBride, J.H. Burton, and R.G. Eggert. 1990.

Health and reproductive performance of dairy cows treated

up to two consecutive lactations with bovine somatotropin.

J. Dairy. Sci. 73 3258.

92

Burton, M.I., B.W. McBride, E. Block, D.R. Glimm, and J.J. Kennelly.

1994. A review of bovine growth hormone. Can. J. Anim.

Sci. 74 167.

Bush, B.M. 1991. Interpretation of Result in Clinical Medicine.

Blackwell Scientif ic Publications, Oxford, UK. Pp. 35.

Capuco, A.V., D.L. Wood, T.H. Elsasser, S. Kahl, R.A. Erdman, C.P.

Vantassel, A. Lefcourt, and L.S. Piperova. 2001. Effect of

bovine somatotropic hormone on thyroid hormones and

cytokines in lactating dairy cows during ad-l ibitum and

restricted feed intake. J. Dairy Sci. 84 2430.

Chaiyabutr,N.,S.Thammacharoen,S.Komolvanich,and S. Chanpongsang.

2005. Effects of long-term administration of recombinant

bovine somatotropin on milk production and plasma insulin-

like growth factor and insulin in crossbred Holstein cows. J.

Agri. Sci. 143 311.

Chalupa, W., and D.T. Gallian. 1989. Nutrit ional implications of

Somatrotropin for lacting cows. J. Dairy Sci. 72 2510.

Chalupa, W., B. Vecchiarelli , D.T. Galligan, J.D. Ferguson, L.S. Baird,

R.W. Hemken, R.J. Harmon, C.G. Soderholm, D.E. Otterby,

R.J. Annexstad, J.G. Linn, P. Hansen, F.R. Ehle, D.L.

Palmquist, and R.G. Eggert. 1996. Responses of dairy cows

supplemented with somatotropin during weeks 5 to 43 of

lactation. J. Dairy Sci. 79 800.

Chill iard, Y. 1989. “Long-term effects of recombinant bovine

somatotropic hormone (rBST) on dairy cow performances A

93

review”. Pp. 61. in K. Sejrsen, M. Vestergaard, and A.

Neimann-Sorensen (eds). Use of Bovine Somatotropic

Hormone in Livestock Production. New York, Ny Elsevier

Applied Sciences.

Clemmons, D.R., and L.E. Underwood. 1991. Nutritional regulation of

IGF-1 and IGF binding proteins. Annu. Rev. Nutr. 11 393.

Collier, R.J., J.C. Byatt, S.C. Denham, P.J. Eppard, A.C. Fabellar, R.L.

Hintz, M.F. McGrath, C.L. McLaughlin, J.K. Shearer, J.J.

Veenhuizen, and J.L. Vicini. 2001. Effects of sustained

release bovine somatotropic hormone (sometribove) on

animal health in commercial dairy herds. J. Dairy Sci. 84

1098.

Dohoo, I.R., L. DesCoteaux, K. Leslie, A. Fredeen, W. Shewfelt, A.

Preston, and P. Dowling. 2003. A meta analysis review of the

effects of recombinant bovine somatotropin. 2. Effects on

animal health, reproductive performance, and culling. Can. J.

Vet. Res. 67 252.

Drackley, J.K., T.M. Cicela, and D.W. LaCount. 2003. Responses of

primiparous and multiparous Holstein cows to addit ional

energy from fat or concentrate during summer. J. Dairy Sci.

86 1306.

Eppard, P.J., D.E. Bauman, and S.N. McWebeoe. 1985. Effect of dose

of bovine growth hormone on lactation of dairy cows. J.

Dairy Sci. 68 1109.

94

Eppard, P.J., D.E. Bauman, C.R.Curtis, H.N. Erb, G.M. Lanza, and M.J.

DeGeeter. 1987. Effect of 188 day treatment with

somatotropin on health and reproductive performance of

lactating dairy cows. J. Dairy Sci. 70 582.

Eppard, P.D., S. Hudson, W.I. Cole, R.L. Hintz, G.F. Hartnell, T.W.

Hunter, L.E. Metzger, A.R. Torkelson, and R.J. Collier.

1991. Response of dairy cows to high doses of a sustained-

release bovine somatotropin administered during two

lactations. 1. Production response. J. Dairy Sci. 74 3807.

Eppard, P.J., L.A. Bentle, B.N. Violand, S.Ganguli, R.L. Hintz, L.

Kung. G.G. Krivi, and G.M. Lanza. 1992. Comparison of the

galactopoietic response to pituitary-derived and recombinant-

derived variants of bovine growth hormone. J. Endol. 132

47.

Eppard, P.J., T.C. White, R.H. Sorbet, M.G. Weiser, W.J. Cole, G.F.

Hartnell, R.L. Intz, G.M. Lanza, J.L. Vicini, and R.J. Collier.

1997. Effect of exogenous somatotropin on hematological

variables of lactating cows and their offspring. J. Dairy Sci.

80 1582.

Esteban, E., P.H. Kass, L.D. Weaver, J.D. Rowe, C.A. Holmberg, C.E.

Franti, and H.F. Troutt. 1994. Interval from calving to

conception in high producing dairy cows treated with

recombinant bovine somatotropin. J. Dairy Sci. 77 2549.

Esteban, E, P.H. Kass, L.D. Weaver, J.D. Rowe, C.A. Holmberg, C.E.

Franti, and H.F. Troutt. 1994. Pregnancy incidence in high

95

producing dairy cows treated with recombinant bovine

somatotropin. J. Dairy Sci. 77 468.

FAO. 1986a. Determination of milk fat by Gerber method Manual of

Food Quality Control. 14/8, P. 8.

FAO. 1986b. Determination of total solids (Rapid method) milk Manual

of Food Quality Control. 14/8, p. 12.

FAO. 2003. FAOSTAT Agriculture Data . Food and Agriculture

Organization Statistics, Rome, Italy.

FAO. 2004. FAOSTAT Agriculture Data . Food and Agriculture

Organization Statistics, Rome, Italy.

Faulkner, A., 1999. Changes in plasma and milk concentration of

glucose and IGF-1 in response to exogenous growth hormone

in lactating goat. J. Dairy Res. 66 207

Gallo, G. F., and E. Block 1990. Effects of recombinant bovine

smotrotropin on nutrit ional status and l iver function of

lactating diary cows. J. Dairy Sci. 73 3276.

Guillermo F., G.F. Gallo, and E. Block. 1990. Effects of recombinant

bovine somatotropin on nutrit ional status and liver function

of lactating dairy cows. J. Dairy Sci. 73 3276.

Gulay, M.S., M.J. Hayen, M. Liboni, T.I. Belloso, C.J. Wilcox, and

H.H. Head. 2004. Low doses of bovine somatotropin during

the transit ion period and early lactation improves milk yield,

eff iciency of production, and other physiological responses

of Holstein cows. J. Dairy Sci. 87 948.

96

Gulay, M.F., and F.H. Hatpolu. 2005. Use of bovine somatotropin in

the management of transit ion dairy cows. Turk. J. Vet. Anim.

Sci. 29 571.

Hansen W.P., D.E. Otterby, J.G. Linn, J.F. Anderson, and R.G. Eggert.

1994. Multifarm use of bovine somatotropin for two

consecutive lactations and its effects on lactational

performance, health, and reproduction. J. Dairy Sci. 7794.

Hard, D.L., W.J. Cole, S.E. Franson, W.A. Samuels, D.E. Bauman, H.N.

Erb, J.T. Huber, and R.C. Lamb. 1988. Effect of long term

sometribove, USAN (recombinant methionly bovine

somatotropin) treatment in a prolonged release system on

milk yield, animal health and reproductive performance-

pooled across four sites. J. Dairy Sci. 71(Suppl)210.

Hatrnell, G.F., S.E. Franson, D.E. Bauman, H.H. Head, J.T. Huber,

R.C. Lamb, K.S. Madsen, W.J. Cole, and R.L. Hintz. 1991.

Long term evalution of sometribove, recombinant methionyl

bovine smatotropin, in a prolonged release system in

lactating diary cows-production responses in a dose titration

study. J. dairy Sci. 74 2645.

Hedge, G.A., H.D Colby, and R.L. Goodman, 1987. Clinical Endocrine

Physiology. W.B. Saunders Co., Philadelphia, P. 69.

Helal, F.I.S., and M.A. Lasheen. 2008. The productive performance of

Egyptian dairy buffaloes receiving biosynthetic bovine

somatotropin (rbST) with or without monensin. American-

Eurasian J. Agric. & Environ. Sci. 3 771.

97

Hemken, R.W., R.J. Harmon, W.J. Sil i iva, W.B. Trucker, G. Heersche,

and R.G. Erregt. 1991. Effect of dietary energy and previous

bovine somatotropin on milk yield, mastit is and reproduction

in dairy cows. J. Dairy Sci. 74 4265.

Hoeben, D., C. Burvenich, P.J. Eppard, and D.L. Hard. 1999. Effect of

recombinant bovine somatotropin on milk production and

composition of cows with Streptococcus uberis mastitis. J.

Dairy Sci. 82 1671.

Holzer, Z., Y, Aharoni, A. Brosh, A. Orlov, J. J. Veenhuizen, and T.R.

Kassar. 1999. The effect of long-term adminstration of

recombinant bovine somatotropin and synovex on

performance, plama hormone and amino acid concentration

and muscle and subcutaneous fat fatty acid composition in

Holstein Friesian bull calves. J. Anim.Sci. 7 1422.

Huber, J.T., Z. Wu, J.R. Fontes, J.L. Sull ivan, R.G. Hoffman, and G.F.

Hartnell. 1997. Administration of recombinant bovine

somatotropin to dairy cows in four consecutive lactations. J.

Dairy Sci. 80 2355.

Hutjens, M.F. 1990. Nutrit ional management economical aspects of

BST. P13. in symposium of 15th Annual Food Animal

Medical Conferance.October 16-17, 1989, Columbus, Ohio.

Jones, B.L. 2000. Economic consequences of extending the calving

intervals of dairy cows treated with recombinant bovine

somatotropic hormone. Working paper by Bruce L. Jones

Professor of Agriculture and Applied Economics, University

of Wisconsin, USA.

98

Jorge, M. A., M. I. F. V. Gomes and R.C. Halt. 2002. Effect of

recombinant bovine omatotropin (bST) uti l ization on milk

production from buffaloes. R. Bras. Zootec. 31 213.

Jousan, F.D., L.A. De Castro e Paula, J. Block, and P.J. Hansen. 2007.

Ferti l i ty of lactating dairy cows administered recombinant

bovine somatotropin during heat stress. J. Dairy Sci. 90 341.

Judge, L.J., P.C. Bartlett, J.W. Lloyd, and R.J. Erskine. 1999.

Recombinant bovine somatotropin association with

reproductive performance in dairy cows. Therio. 52 481.

Judge, L.J., J.W. Lloyd, and P.C. Bartlett. 2009. The Effect of

recombinant bovine somatotropin on patterns of milk

production, lactational milk estimates and net farm income.

NEP- AGR. 10-24 (Agricultural Economics).

Juskevich, J.C., and C.G. Guyer. 1990. Bovine growth hormone. Human

Food Safety Evaluation. Sci. P. 249.

Khattab, H. M. M., H. M. El-Sayed, S.A.H. Abo El-Nor, H.A. EL-

Alamy and R. M. A. Abd El-Gawad. 2008. Impact of bovine

somatotropin and monensin on the productive performance of

Egyptian dairy buffaloes. Intl. J. Dairy Sci. 3 11.

Kinoshita,J., N. Suzuki, and H.M. Kaiser. 2004. An economic

evaluation of recombinant bovine somatotropin approval in

Japan. J. Dairy Sci.87 1565.

Kirchgessner, M., W. Windisch, W. Schwab, and H.L. Muller. 1991.

Energy metabolism of lactating dairy cows treated with

99

prolonged-release bovine somatotropic hormone or energy

deficiency. J. Dairy Sci. 74 (Suppl.2) 35.

Kostyo, J. L., and R.C. Reagan. 1976. The biology of somatotropin.

Pharmacol. Theriogenol. 2 591.

Kweon, U.G.K.T., H.S. Nam, S.K. Kim, J.S. Yun, and. B.J. Hang, 2000.

Effect of recombinant bovine somatotrpic hormone injection

time on haemotological values and blood metabolic

substrates in Holstein steers. Kor .J. Anim. Sci. 42 269.

Lanza G.M., T.C. White, S.E. Dyer, S. Hudson, S.E. Franson, R.L

Hintz, J.A. Duque S.C. Bussen, R.K. Leak, and L.E.

Metzger. 1988. Response of lactating dairy cows to

intramuscular or subcutaneous injection of sometribove,

USAN (recombinant methionyl bovine somatotropin) in a

14-day prolonged release system. Part II. Changes in

circulating analytes. J. Dairy Sci. 71 (suppl. 1) 195.

Lormore, M. J., L.D. Muller, D.R. Deaver, and L.C. Griel. 1990. Early

lactation responses in dairy cows administered bovine

somatotropin and fed diets high in energy and protein. J. Dairy

Sci. 733237.

Lean, I.J., R. L. Baldwin, H.F. Troutt, M.L. Bruss, J.C. Galland, T.B

Farver, J.Rostami, L.D. Weaver, and C.A. Holmberg. 1992.

Impact of bovine somatotropin administration beginning at

day 70 of lactation on serum metabolites, milk constituents,

and production in cows previously exposed to exogenous

somatotropin. Anim. J. Vet, Res. 53 731.

Leonard, M.,and E. Block. 1997. Effects on nutrient and hormonal

profi le of long-term infusions of glucose or insulin plus

100

glucose in cows treated with recombinant bovine

somatotropin before peak milk yield. J. Dairy Sci. 80 127.

Lesniak, M.A., P. Gorden, and J.Roth. 1977. Reactivity of non-primate

growth hormones and prolactins with human growth hormone

receptors on cultured human lymphocytes. J. Clin. End.

Metab. 44 838.

Linn, J. G. 1987. Factors effecting the composition of milk from dairy

cows. In Designing Foods Animal Product Options in the

Marketplace. National Academic Press. Washington, DC., Pp.

224.

Lough D.S., L.D. Muller, R.D. Kensinger, L.C. Griel, Jr., and C.D.

Azzara. 1989. Effect of Exogenous bovine somatotropin on

mammary lipid metabolism and milk yield in lactating dairy

cows. J Dairy Sci 721469.

Lucy, M. C., S.D. Hauser, P.J. Eppard, G.G. Krivi, J.H. Clark, D.E.

Bauman, and R.J. Collier. 1993. Variants of somatotropin in

cattle gene frequencies in major dairy breeds and associated

milk production. Domest. Anim. End. 10 325

Ludri, R.S., R.C. Upadhyay, and Mahendra. 1989. Milk production in

lactating buffalo receiving reconbinantly produced bovine

somatotropic hormone. J. Dairy Sci. 72 2283.

Luna- Dominguez, E.L., R.M. Enns D.V. Armstrong and R.L. Ax. 2000.

Reproductive performance of Holstein cows receiving

somatotropin. J. Dairy Sci. 83 1451.

101

Lynch, J.M., D.M. Barbano, D.E. Bauman, and G.F. Hartnel. 1988.

Influence of sometribove (recombinant methinoyl bovine

somatotropin) on the protein and fatty acid composition of

milk. J. Dairy Sci. 71(Suppl.1)100.

Lynch, J.M., D.M. Barbano, D.E. Bauman, G.F. Hartnell, and M. A.

Nemeth. 1992. Effect of a prolonged-release formulation of

N-methionyl bovine somatotropin (sometribove) on milk fat.

J. Dairy Sci. 75 1794.

Mandal, A.B., S.S. Paual, and N.N. Pathak. 2003. Nutrient requirements

and feeding of buffaloes and cattle. Published by Int. Book

distributing Co. Charbagh, Lucknow, India.

McBride, B.W., J.L. Burton, J.H. Burton, G.K. Meleod, and R. Eggert.

1989. Multi lactational treatment effects of rbST on

production responses in lactating Holstein cows. J. Dairy Sci.

72(Suppl.1) 430.

McBride, B.W., W. D. Short, S. El-Osta, and Hisham. 2002. Production

and financial impacts of the adoption of bovine somatotropin

on U.S. dairy farms. 2002 Annual meeting, July 28-31, Long

Beach, CA with number 19908 American Agricultural

Economics Association.

McGuffey, R.K., H.B. Green, R.P. Basson, and T.H. Ferguson. 1990.

Lactation response of dairy cows receiving bovine

somatotropic hormone via daily injections or in a sustained-

release vehicle. J. Dairy Sci. 73763.

102

McGuffey, R.K., and J.I. D. Wilkinson. 1991a. Nutrit ional implications

of bovine somatotropin for the slactating dairy cow. J. Dairy

Sci. 74, Suppl. 2 63.

McGuffey, R.k., R. P. Basson, and T.E. Spike. 1991b. Lactation

response and body composition of cows receiving

somatotropin and three ratios of forage to concentrate. J.

Dairy Sci. 74 3095.

McGuffey, R.K., R.P. Basson, D.L. Snyder, E. Block, J.H. Harrison,

A.H. Rakes, R.S. Emery, and L.D. Muller. 1991c. Effect of

somidobove sustained release administration on the lactation

performance of dairy cows. J. Dairy Sci. 741263.

McGuire M.A., D.E. Bauman, M.A. Miller, and G.F. Hartnell. 1992.

Response of somatomedins (IGF-I and IGF-II) in lactating

cows to variations in dietary energy and protein and

treatment with recombinant n-methionyl bovine somatotropin.

J. Nutr. 122.

Mehra, K.L. 2001. Animal biotechnologies; benefits and concerns. Nat.

Acad. Agric. Sci. New Delhi-110067, India.

Mellado, M., E. Nazarre, L. Olivares, F. Pastor, and A. Estrada. 2006.

Milk production and reproductive performance of cows

induced into lactation and treated with bovine somatotropin.

J. Anim. Sci. 82 555.

Miller, P.S., B.L. Reis, C.C. Calvert, E.J. De Peters, and R.L. Baldwin.

1991. Patterns of nutrient uptake by the mammary glands of

lactating dairy cows. J. Dairy Sci. 74 3791.

103

Miller, A.R.E., E. P. Stanisiewski, R.A. Erdman, L.W. Douglas, and

G.E. Dahl. 1999. Effect of long daily photoperiod and bovine

somatotropin (Trobest) on milk yield in cows. J. Dairy Sci.

82 1716.

Ministry of Food and Economics and Statistics. 2006. Livestock Census

2006. Ministry of Food and Economics. & Statistic,

Pakistan.

MINITAB. 1996. Manual Release II. Minitab Inc. State Coll. PA.

Mishra, A., and D.C. Shukla. 2004. Effect of recombinant bovine

somatotropic hormone (Boostin-250) on blood metabolites

and milk yield of lactating buffaloes. Asian-Aust. J. Anim.

Sci. 17 1232.

Moallem, U., Y. Folman, and D. Sklan. 2000. Effects of somatotropin

and dietary calcium soaps of fatty acids in early lactation on

milk production, dry matter intake and energy balance of

high-yielding dairy cows. J. Dairy Sci. 83 2085.

Molento, C.F., E. Block, R.I. Cue, and D. Petitclerc. 2002. Effects of

insulin, recombinant bovine somatotropin, and their

interaction on insulin-like growth factor-I secretion and milk

protein production in dairy cows. J. Dairy Sci. 85 738.

Mollet, T.A., M.J. De Geeter, R.L. Belyea, R.A. Youngquist, and G.M.

Lanza. 1986. Biosynthetic or Pituitary extracted growth

hormone induc. glactopoiesis in dairy cows. J. Dairy Sci. 69

(Suppl. 1) 118.

104

Moreira, F., C. A. Risco, M. F. Pires, J. D. Ambrose, M. Drost, and W.

W. Thatcher. 2000. Use of bovine somatotropin in lactating

dairy cows receiving timed artif icial insemination. J. Dairy

Sci. 83 1237.

Muhammad, G., M. Akthar, A. Shakoor, M.Z. Khan, F. Rehman, and

M.T. Ahmad. 1995. Surf f ield mastitis test an expensive

new tool for evaluation of wholesomeness of fresh milk.

Pak .Vet. J. 5 3

National Research Council (NRC). 1988. Metabolic Modif ier- Effect on

the Nutrient Requirement of Food Producing Animals. National

Academy Press, Washington, DC.

National Research Council (NRC). 1989. National Academy Press,

Washington, DC.

National Research Council (NRC). 1994. Metabolic Modif ier- Effect on

the Nutrient Requirement of Food Producing Animals. National

Academy Press, Washington, DC.

National Research Council (NRC). 2001. Nutrient Requirements of

Dairy Cattle. Natl. Acad. Sci. Press. Washington, DC.

Newbodl, J.A., R.B. Heap, C.G. Prosser, R.H. Phipps, F. Adrianes and

D.L. Hard. 1997. The effect of bovine somatotropic hormone

and diet on Bovine somatotropic hormone binding sites in

hepatic tissue of lactating dairy cows. J. Dairy Sci. 80 1085.

Nisa, M., A. Sufyan, M. Sarwar, and M. A. Shahzad. 2006. Influence of

bovine somatotropin and varying levels of enzose on

nutrients intake, digestibi l i ty, milk yield and its composition

in mid-lactating Nili-Ravi buffaloes. J. Dairy Sci. 89 1157.

105

Otto, G., M. Khalid, and H. Torsten. 2003. A review of milk production

in Pakistan with particular emphasis on small-scale

production. PPLPI Working Paper No. 3. Pro-Poor Livestock

Policy Initiative. Food and Agricultural Organization, Animal

Production and Health Division, Rome, Italy.

Patton, R.A., and C.W. Healed. 1992. Management of bST

supplemented cows. Bovine somatropin and emerging issues

edited by M.C. Hallberg. Boulder, Co. Westview. P. 73- 98.

Peel, C. J., and D. E. Bauman. 1987. Somatotropin and lactation. J.

Dairy Sci. 70 474.

Peel, C. J., D.L. Hard, K.S. Madsen, and G. de Kerchove. 1989.

Monsanto technical symposium Pioc. Cornell Nutrition

Conference. Anim Sci. Div. Monsanto Agriculture Co. St.

Louis, Mo.

Peters, J. P. 1986. Consequances of accelerated gain and growth

hormone administration for lipid metabolism in growing

beef steers. J. Nutr. 116 2490.

Phipps, R.H., D.L. Hard, and F. Adriaens. 1997. Use of bovine

somatotropin in the tropics. The effect of sometribove on

milk production in Western, Eastern, and Southern Africa. J.

Dairy Sci. 80 504.

Polidori F, R. C.A. Sgoifo, E.M. Senatore, G. Savoini, and V.

Dell 'Orto. 1997. Effect of recombinant bovine somatotropin

and calcium salts of long-chain fatty acids on milk from

Italian buffalo. J. Dairy Sci. 80 2137.

106

Rechler, M.M., and S.P. Nissley. 1990. Insulin-l ike growth factors.

Handb. Exp. Pharmacol. 95 263.

Remond, B., M. Cisse, A. Ollier, and Y. Chill iard. 1991. Slow release

somatotropin in dairy heifers and cows fed two levels of

energy concentrate for performance and body condition. J.

Dairy Sci. 74 1370.

Richard, R.O., and A.K. Davis. 1947. The application of f lame

photometery to sodium and potassium determination in

biological f luids. J.Biol.Chem.171 641.

Robinson, P.H., G. DeBaer and J.J. Kennelly, 1991. Effect of bovine

samatotropin and protein on ruman fermentation and

forestomach and whole tract digestion in dairy cows. J. Dairy

Sci. 743505.

Rose, M.T., T.E.C. Weekes, and P. Rowlinson. 2004. Individual

variation in the milk yield response to bovine somatotropic

hormone in dairy cows. J. Dairy Sci. 87 2024.

Roush, W. 1991. Who decides about biotech the clash over bovine

growth hormone. Tech. Rev. 14 28.

Rowlands, G.J., W. Little, A.J. Stark and R. Manston. 1979. The blood

composition of cows in commercial dairy herds and its

relationship with season and lactation. Proceedings of

International Conference on Production Disease in Farm

Animals. 7 324.

Santos, J.E., J.T. Huber, C.B. Theurer, L.G. Nussio, C.B. Nussio, M.

Tarazon, and R.O. Lima-Filho. 1999. Performance and

107

nutrient digestibil i ty by diary cows treated with bovine

somatotropic hormone and fed diets with steam flaved

sorghum or steam rolled corn during early lactation. J. Dairy

Sci.82 404.

Santos, J.E.P., S.O. Juchem, R.L.A. Cerri, K.N. Galva, R.C. Chebel,

W.W. Thatcher, C.S. Dei, and C.R. Bilby. 2004. Effect of

bST on reproductive management and reproductive

performance of Holstein dairy cows. J. Dairy Sci. 87 868.

Sarwar, M. M.A. Shahzad, M. Nisa, and M.A. Sufyan. 2007. Influence

of bovine somatotropin and replacement of corn dextrose

with concentrate on the performance of mid-lactating

buffaloes fed urea-treated wheat straw. Turk. J. Vet. Anim.

Sci. 31 259.

Sastry, A.G. 1983. Veterinary Clinical Pathology. (3rd ed.)

Philadelphia, PA lea and Febiger.

Schams, O.W., N.C. Jain and E.J. Carroll. 1975. Veterinary

Hematology, (3rd ed.) Philadelphia, PA lea and Febiger.

Schams, D., F. Graf, J. Meyer, B. Graule, M. Mauthner, and C.

Wollny. 1991. Changes in hormones, metabolites, and milk

after treatment with sometribove (recombinant methionyl

bST) in Deutsches Fleckvieh and German black and white

cows. J. Dairy Sci. 69 1583.

Silvia, W.J., R.W. Hemkenand T.B. Hatler. 2002. Timing of onset of

somatotropin supplementation on reproductive performance

in dairy cows. J. Dairy Sci. 85 384.

108

Skarda. J., E. Markalous, J. Slaba, P. Krejci, O. Skardova, and J.

Zednik. 1992. Effect of methionyl bovine somatotropin in a

prolonged-release vehicle on milk production, hormone

profi les and health in dairy cows. J. Dairy Sci. 79 499.

Slaba. J., P. Krejci, J. Skarda, L.M. Huybrechts, E. Decuypere, and H.

Herrmann. 2005. Plasma profiles of somatotropin and IGF-I

in diary cows following application of two preparations of

recombinant bovine somatotropin in a sustained release

vehicle. J. Dairy Sci. 8 37.

Solderhalm, G.G., D.E. Otterby, J.G. Linn, F.R. Ehle, G.E. Wheaton,

W.P. Hansen, and R.J. Annexstad. 1988. Effects of

recombinant bovine somatotropic hormone on milk

production, body composit ion, and physiological parameters.

J. Dairy Sci. 71 355.

Spicer, L. J., W. B. Tucjer, and G. Dadams. 1990 Insulin-l ike growth

factor-I in diary cows; relationships among energy balance,

body condition, ovarian activity, and estrous behavior. J.

Dairy Sci. 73929.

Srinivasa-Rao, K., and M. Ranganadham. 2000. Effect of bovine

somatotropin on milk production and composit ion in

lactating Murrah buffaloes. Indian J. Dairy Sci. 53 46.

Srivastava, A., and V.D. Mudgal. 1984. Effect of feeding protected fat

to buffaloes on milk production. Indian J. Anim. Sci.54

1122.

109

Starbuck, M. J., E. K. Inskeep, and R. A. Dailey. 2006. Effect of a

single growth hormone (rbST) treatment at breeding on

conception rates and pregnancy retention in dairy and beef

cattle. Anim. Reprod. Sci. 93 349.

Stegeman G. A., R. J. Baer, D. J. Schingoethe, and D. P. Casper. 1992.

Composit ion and flavor of milk and butter from cows fed

unsaturated dietary fat and receiving bovine somatotropin. J.

Dairy Sci. 75 962.

Steel, R. G. D., J.H. Torrie, and D.A. Dickey. 1997. Principles and

Procedures of Statistics. Biochemical Approach (3rd

ed.).McGraw Hil l Book Co. Inc., New York, USA.

Tarazon-Herrera, M. A., J. T. Huber, J. E.P. Santos, and L.G. Nussio.

2000. Effects of bovine somatotropin on milk yield and

composition in Holstein cows in advanced lactation fed low-

or high-energy diets. J. Dairy Sci. 83 430.

Tauer, L.W., and W.A. Knoblauch. 1997. The empirical impact of

bovine somatotropin on New York dairy farms. J. Dairy Sci.

80 1092.

Tauer, L.W. 2008. Estimatiom of treatments effects of recombinants

bovine somatotropin using matching samples. Working paper

51118, Cornell Univesity Department of Applied Economics

and Management.

Tessman, N. J., T. R. Dhiman, J. Klenmanns, H. D. Radloff, and L.D.

Satter. 1991. Recombinant bovine somatotropin with

110

lactating cows fed diets differing in energy density. J. Dairy

Sci. 74 2633.

Thatcher, W.W., T.R. Bilby, J.A. Bartolome, F. Silvestre, C.R. Staple,

and J.E.P. Santos. 2006. Strategies for improving ferti l ity in

the modern dairy cow. Therio. 65 30.

Thomas, J.W., W.A. Samuels, and C.J. Peel. 1990. Evaluation of

management practices and sometribove, USAN

(recombinant methionyl) bovine somatotropin in Michigan

dairy herds during two years. J. Dairy Sci. 73 158.

Tuggle, C.K., and A. Trenkle. 1996. Control of growth hormone

synthesis. Domest. Anin. Endocrinol. 13 1.

Usmani, R.U., A.G. Khan, and I.H. Athar. 1991. A review of bovine

growth hormone biotechnology for increased milk

production. Pak. Vet. J. 11 100.

Vallimont, J.E., G.A. Varga, A. Ariel i, T.W. Cassidy and K.A.

Cummins. 2001. Effect of prepartum somatotropin and

monansin metabolisim, production of beef cattle. J. Ji l in.

Agri. Uni. 21 5.

VanBaale, M.J., D.R. Ledwith, J.M. Thompson, R. Burgos, R.J. Coll ier,

and L.H. Baumgard. 2005. Effect of increased milking

frequency in early lactation with or without recombinant

bovine somatotropin. J. Dairy Sci. 88 3905.

111

Van Soest, P.J., H.B. Robertson, and B.A. Lewis. 1991. Methods of

dietary f iber, NDF and non-starch- polysaccharides in

relation to animal material. J. Dairy Sci.74 3583.

Vicini, J.L., G.F. Hartnell, J.J. Veenhuizen, R.J. Collier, and L.

Munyakazi. 1994. Effect of supplemental dietary fat or

protein on the short-term milk production response to bovine

somatotropin. J. Dairy Sci. 78 863.

Wallis, M. 1975. The molecular evolution of pituitary hormones. Biol.

Rev. 50 35.

Waterman, D.F., W.J. Sil ivia, R.W. Hemken, G.J. Heersche, T S.

Swenson, and R.G. Eggert. 1993. Effect of bovine

somatotropin on reproductive function in lactating dairy

cows. Therio. 40 1015.

White, T.C., K. S. Madsen, R.L. Hinz, R.H. Sorbet, R.J. Coll ier, D.L.

Hard, G.F. Hartnell, G.D.E. Kerchove, F. Adriaens, N.

Craven, D.E. Bauman, G. Bertrand, P.H. Bruneau, G.O.

Gravert, H.H. Head, J.T. Huber, R.C. Lam, C. Palmer, A.N.

Pell, R. Phipps, R. Weller, G. Piva, Y. Ripjkema, J. Skarda,

J. Vedeau, and C. Wollny. 1994. Clinical mastit is in cows

treated with sometribove (recombinant bovine bovine

somatotropic hormone) and its relationship to milk yield. J.

Dairy Sci. 77 2249.

Wood, D.C., W.J. Salsgiver, T.R. Kasser, G.W. Lange, E. Rowold, B.N.

Violand, A. Johnson, R.M. Leimgruber, G.R. Parr, N.R.

Siegel, N.M. Kimack, C.E. Smith, J.F. Zobel, S.M. Ganguli,

J.R. Garbow, G. Bild, and G.G. Krivi. 1989. Purif ication and

112

characterization of pituitary bovine somatotropin. J. Biol.

Chem. 264 14741.

Young, F.G. 1947. Experimental stimulation (galactopoiesis) of

lactation. Br. Med. Bull. 5 155.

Zhao, X,. J.H. Burton, B.W. Mc Bride. 1992 Lactation, health, and

reproduction of dairy cows receiving daily injectali le or

sustained release somatotropin. J. Dairy SCI. 753122