by ahmed soliman hassan soliman - puntoganadero.cl

IMPACT OF USING ANTIOXIDANT AGENTS ON REPRODUCTIVE

PERFORMANCE AND THERMOREGULATION IN ABERDEEN ANGUS

COWS UNDER NEW VALLEY CONDITIONS

By AHMED SOLIMAN HASSAN SOLIMAN

B.Sc. Agriculture Science, Faculty Of Agriculture (Animal Production),

New Valley Branch, Assiut University, 2013

A Thesis

Submitted in partial fulfillment of the requirements for the

degree of master of Agricultural Sciences

In

Animal Production - Animal Physiology

Supervision Committee :- Prof. Dr. Hassan Abdel-Ghany Hassan Daghash Professor Of Animal Physiology, Department Of Animal Production, Faculty Of Agriculture, Assiut University Dr. Ayman Youssef Mohamed Ahmed Kassab Associate Professor Of Animal Physiology, Department Of Animal Production, Faculty Of Agriculture, New Valley University Dr. Hatem Abd-Elkader Hamdon Associate Professor Of Animal husbandry, Department Of Animal Production, Faculty Of Agriculture, New Valley University

Dr. Waleed Senosy Ali Senosy Associate professor Of Theriogenology, Faculty Of Veterinary Medicine, New Valley University

2018 A – 1440 H

Assiut University

Faculty Of Agriculture

Department Of Animal Production

ACKNOWLEDGEMENT

Acknowledgements

ACKNOWLEDGMENTS

I express my deep thanks to Allah who fulfilled my hopes and promise to

offer every possible aid for any one need to Allah.

I express my appreciation to Prof. Dr. Hassan A. Dagash Professor of

Animal Physiology, Department of Animal Production, Faculty of Agriculture,

Assiut University for suggesting the current study, supervision, valuable advice,

guidance during this study.

I express my appreciation to Dr. Ayman Y. Kassab Associate Professor

of Animal Physiology, Department of Animal Production, Faculty of

Agriculture New Valley, Assiut University for his efforts in providing assistance

and providing the laboratory and tools required during the study .

I express my appreciation to Dr. Hatem A. Hamdon Associate Professor

of Animal Husbandry , Department of Animal Production, Faculty of

Agriculture New Valley, Assiut University for supervision, valuable advice,

guidance and valuable help during the preparation of this manuscript

I express my appreciation to Dr. Waleed S. A. Senosy Associate

professor of Theriogenology, Faculty of Veterinary Medicine New Valley,

Assiut University for supervision, valuable advice, guidance during this study.

Great thanks for Prof. Dr Mohamed N. Abd El-Ati Professor of Animal

Production , Faculty of Agriculture, Assiut University and Dr. Mostafa G. Abd

El-Fattah Lecturer of Poultry Production , Faculty of Agriculture, Assiut

University for their help in Statistical analyzing

I want to thank my friend Ali Khalifa and Eng. Ghada Khidiwe for their

help in analyzing serum and blood samples for haemato-biochemical

parameters.

Finally, I would like to extend my thanks to my father, mother, sisters

and brother for their support and everyone who gave me a hand to complete

this study.

CONTENTS

Contents

I

CONTENS Page SUBJECT

1 INTRODUCTION 1- 4 REVIEW OF LITERATURE 2- 4 Effects of using antioxidant on thermoregulatory responses 2-1- 6 Effects of using antioxidant on blood hematological parameters 2-2- 6 White blood cells and its fractions 2-2-1 - 8 Red blood cells 2-2-2 - 9 Effects of using antioxidant on blood biochemical parameters 2-3- 9 Blood total protein and its fractions 2-3-1 : 11 Total cholesterol 2-3-2 : 13 Glucose 2-3-3 : 14 Liver function (ALT and AST enzymes ) 2-3-4 : 15 Kidney function ( Urea and creatinine ) 2-3-5 : 17 Total antioxidant capacity (TAC) 2-3-6 : 18 Effects of using antioxidant on reproductive performance 2-4 : 20 Ovarian activity 2-4-1: 21 Conception rate 2-4-2 : 23 Service per conception 2-4-3 : 24 Estrogen and Progesterone hormone 2-3-3 : 26 MATERIALS AND METHODS 3: 28 Meteorological parameters 3-1-1: 28 Thermal responses 3-1-2: 29 Blood sampling and analytical methods 3-1-3: 30 Estrous synchronization using prostaglandins 3-2-1: 31 Trans-rectal ultrasonographic examination 3-2-2: 33 Reproductive parameters of cows 3-2-3: 33 Statistical Analysis 3-3: 34 RESULTS AND DISCUSSION 4- 34 Ambient temperature , relative humidity and temperature-humidity

index (THI) during experimental period . 4-1-

35 Effects of using antioxidant agents on thermoregulatory responses of experimental cows

4-2-

40 Effects of antioxidant on blood hematological parameters of experimental cows

4-3

40 White blood cells and its fractions 2-3-1 : 42 Red blood cells , Hb , HCT 2-3-2 : 45 Effects of antioxidant on blood biochemical parameters of

experimental cows 4-4

45 Total protein 4-4-1 :

Contents

II

47 Albumin 4-4-2 : 48 Globulin 4-4-3 : 49 Glucose 4-2-4 : 50 Total cholesterol 4-4-5 : 51 Liver function (ALT and AST enzymes ) 4-4-6 : 51 Aspartate Amino Trsnsferase (AST) 4-4-6-1 : 53 Alanine Amino Transferase (ALT) 4-4-6-2: 54 Kidney function ( urea-N and creatinine ) 4-4-7 : 55 Total Antioxidant Capacity (TAC) 4-4-8 : 57 Effects of antioxidant on reproductive performances of

experimental cows 4-5

57 Response to synchronization 4-5-1: 58 First service conception rate 4-5-2 : 59 Conception rate 4-5-3 : 61 Number of services per conception 4-5-4 : 62 Size of embryo, amniotic vesicle and corpus luteum 4-5-5 : 64 Ovarian follicular development and hormonal profile during estrous

cycle 4-5-6:

64 Follicular development 4-5-6-1: 69 Concentration of estrogen and progesterone hormones 4-5-6-2: 75 Correlation between ovarian follicular and luteal development and

hormone profile 4-5-6-3:

78 SUMMARY AND CONCLUSION 5- 82 REFERENCES 6- 1 ARABIC SUMMARY

LIST OF TABLES

List of Tables

III

LIST OF TABLES Page TITLE

27 Composition of concentrate feed mixture and treatments . Table (1) 27 Chemical composition of concentrate feed mixture and wheat straw. Table (2) 34 Means of ambient temperature (°C), relative humidity (%) and

temperature humidity index (THI) during the experimental period . Table (3)

36 Effect of treated cows with antioxidant on thermoregulatory responses (mean±SEM)

Table (4)

41 Effect of treated cows with antioxidant on white blood cell (WBC) and its fractions (mean±SEM)

Table (5)

43 Effect of treated cows with antioxidant on red blood cell RBCs , Hb and HCT (mean±SEM)

Table (6)

46 Effect of treated cows with antioxidant on total protein and its fractions (mean±SEM)

Table (7)

50 Effect of treated cows with antioxidant on glucose, cholesterol, AST and ALT (mean±SEM)

Table (8)

54 Effect of treated cows with antioxidant agents on urea-N, creatinine , and total antioxidant capacity (TAC) (mean±SEM)

Table (9)

58 Effect of treated cows with antioxidant on reproductive performance . Table (10) 63 Effect of treated cows with antioxidant on size of embryo (mm),

amniotic vesicle (mm) and corpus luteum. (mean±SEM) Table (11)

65 Effect of treated cows with antioxidant on number of different size of ovarian follicles during estrus period (mean ± SEM)

Table (12)

67 Effect of treated cows with antioxidant on size of largest follicle of Aberdeen Angus cows during estrus period (mean ± SEM)

Table (13)

68 Effect of treated cows with antioxidant on size of corpus lutem of Aberdeen Angus cows during estrus period (mean±SEM)

Table (14)

69 Effect of treated cows with antioxidant on estrogen concentrations during estrous period of Aberdeen Angus cows (mean±SEM)

Table (15)

70 Effect of treated cows with antioxidant on progesterone concentrations during estrous period of Aberdeen Angus cows (mean±SEM)

Table (16)

LIST OF FIGURES

List of Figures

IV

LIST OF FIGURES Page Title

31 Estrous synchronization using prostaglandins Fig. (1) 34 Means of temperature humidity index (THI) during the

experimental period. Fig. (2)

37 Effect of treated cows with antioxidant on rectal temperature, respiration rate and pulse rate of experimental cows.

Fig. (3)

39 Effect of treated cows with antioxidant on skin temperature, hair temperature and ear temperature of experimental cows .

Fig. (4)

41 Effect of treated cows with antioxidant on WBCs and its fractions. Fig. (5) 43 Effect of treated cows with antioxidant on RBC and HB . Fig. (6) 44 Effect of treated cows with antioxidant on HCT, MCV, MCH ,

MCHC and RDW . Fig. (7)

46 Effect of treated cows with antioxidant on total protein, albumin and globulin

Fig. (8)

51 Effect of treated cows with antioxidant on glucose, cholesterol, AST and ALT

Fig. (9)

54 Effect of treated cows with antioxidant on urea-N and creatinine Fig. (10) 55 Effect of treated cows with antioxidant on total antioxidant

capacity (TAC) Fig. (11)

58 Effect of treated cows with antioxidant on first service conception rate

Fig. (12)

59 Effect of treated cows with antioxidant on conception rate Fig. (13) 61 Effect of treated cows with antioxidant on number of services per

conception. Fig. (14)

63 Effect of treated cows with antioxidant agents on size of embryo(mm) and amniotic vesicle (mm)

Fig. (15)

66 Effect of treated cows with antioxidant on number of different size of ovarian follicles of Aberdeen Angus cows during estrus period

Fig. (16)

69 Effect of treated cows with antioxidant on estrogen concentrations during estrous period of Aberdeen Angus cows

Fig. (17)

71 Effect of treated cows with antioxidant on progesterone concentrations during estrous period of Aberdeen Angus cows

Fig. (18)

72 Estrogen and progesterone concentrations during estrous period of Aberdeen Angus cows

Fig. (19)

75 A positive correlation between progesterone concentrations and corpus luteum during estrous period of Aberdeen Angus cows

Fig. (20)

75 Ultrasonographic view from ovaries with non-vacuolated CL Fig. (21) 76 Ultrasonographic view from ovaries with vacuolated CL Fig. (22) 76 A positive correlation between estrogen concentrations and largest

follicle during estrous period of Aberdeen Angus cows Fig. (23)

List of Figures

V

77 Ultrasonographic view from ovaries with small follicle (< 5 mm) Fig. (24) 77 Ultrasonographic view from ovaries with follicle ( 5-12 mm) Fig. (25) 77 Ultrasonographic view from ovaries with ovulatory follicle ( > 12

mm) Fig. (26)

LIST OF

ABBREVIATIONS

List of Abbreviations

V

LIST OF ABBREVIATIONS

% Percentage

°C Degree Celsius

ALT Alanine aminotransferase

AST Aspartate aminotransferase

Ca Calcium

CF Crude Fiber

CL Corpus luteum

CP Crude Protein

Cu Copper

DM Dry Matter

EDTA Ethylenediaminetetraacetic acid

ET Ear temperature

GRAN Granulocytes

GSH-Px Glutathione peroxidase

Hb Hemoglobin concentration

HCT Hematocrit

HT Hair temperature

I Iodine

LYM Lymphocytes

MCH Mean corpuscular hemoglobin

MCHC Mean corpuscular hemoglobin concentration

MCV Mean corpuscular volume

Mg Magnesium

MID Monocytes

List of Abbreviations

VI

Mn Manganese

NFE Nitrogen Free Extract

P Phosphorous

PG Prostaglandin

PR Pulse rate

RBC Red blood cell

RDW Red Blood Cell Distribution Width

RR Respiration rate

RT Rectal temperature

Se Selenium

SOD Superoxide dismutase

ST Skin temperature

TAC Total antioxidant capacity

THI Temperature humidity index

WBC White blood cell

Zn Zinc

INTRODUCTION

Introduction

1

1-INTRODUCTION

The Aberdeen Angus is a Scottish breed of small beef cattle. It derives

from cattle native to the counties of Aberdeenshire and Angus in north-eastern

Scotland. The main use of Angus cattle is for beef production and consumption.

The beef can be marketed as superior due to its marbled appearance. This has

led to many markets, including Australia, Japan and the United Kingdom to

adopt it into the mainstream.( www.wikipedia.com )

Heat stress induces oxidative stress and reduces antioxidant status by

promoting overproduction of free radicals and reactive oxygen species and by impairing the antioxidant defense system (Alhidary et al., 2015 and Altan et

al., 2000). Hot environmental conditions markedly decrease blood concentration

of antioxidant micronutrients (Zn, Se, and vitamin E) in ruminants (Burke et al.,

2007 and Saker et al., 2004 ) and poultry (Saini et al., 2007 and Altan et al.,

2003 ) . Moreover, Oxidative stress plays a major part in the development of

chronic and degenerative ailments such as cancer, autoimmune disorders,

rheumatoid arthritis, cataract, aging, cardiovascular and neurodegenerative

diseases ( Pham-Huy et al ., 2008) .

Minerals and vitamins play an important role in the growth and

reproductive performance of farm animals. Vitamin E and the trace elements

selenium (Se) and zinc (Zn) are essential for the health and performance of beef

cows. These micronutrients are cellular antioxidants, preventing peroxidative

damage, either in cell membranes (vitamins) or in the cytoplasm (trace

elements), and are essential for a well functioning immune system. ( Weiss,

2002 ).

Minerals such as phosphorous (P), calcium (Ca), magnesium (Mg), iodine

(I), manganese (Mn), copper (Cu), selenium (Se), and zinc (Zn) are all involved

in governing successful reproductive processes ( Wilde, 2006 ). Some trace

elements, such as Cu, Zn, Se and Mo are involved in cellular respiration, cellular

Introduction

2

utilization of oxygen, DNA and RNA replication, maintenance of cell membrane

integrity, and sequestration of free radicals ( Chan et al., 1998 )

The most important biological antioxidants are vitamins A, C, E and

selenium, a key component of glutathione peroxidase, Vitamin E is an important

lipid soluble antioxidant that protects against free radical-initiated lipid

peroxidation ( NRC ., 2001 ). However, the amount of vitamin E and selenium

needed for maximizing immune competence is higher than the suggested

requirements of NRC (Nockels, 1996) .

The mode of action of vitamin E is closely associated with Se metabolism, and it can make up for selenium deficiency to a certain degree (Makimura et

al., 1993; Soliman 2015 b ) . Vitamin E supplementation (LeBlanc et al., 2004

and Panda et al., 2006) improves reproductive performance because of their

positive effect on steroid synthesis, release, follicular growth and symptoms of

ovulatory oestrus (Srivastava, 2008).

Several studies have demonstrated interaction between minerals and

reproduction in ruminant . For example, flushing or minerals improves has been

shown to improve production and reproduction parameters (Madibela et al.,

2002; Fernandez et al.,2004; Almeida et al., 2007; Griffiths et al., 2007).

Zinc is an essential component of numerous enzymes including enzymes

involved in the synthesis of DNA and RNA. In the antioxidant system Zn is a

component of Cu–Zn Superoxide dismutase (SOD) . Zinc also induces synthesis

of metallo - thionein, a metal binding protein that may scavenge hydroxide radicals ( Prasad et al., 2004) . In addition to an antioxidant role, Zn may affect

immunity via its important role in cell replication and proliferation (Weiss and

Spears, 2006).

As integral parts of the antioxidant defense system, Selenium and vitamin

E play important roles in the growth and health of humans and animals (Surai,

Introduction

3

2006 and McDowell, 2003). These micronutrients are often added to diets to

improve livestock production and enhance immune competence. The addition

of selenium and vitamin E improved glutathione peroxidase (GSH-Px) activity

in blood and increases plasma concentrations of total antioxidant capacity and selenium in dairy cattle (Calamari et al., 2011) and goats (Katamoto et al.,

1998).

Chauhan et al ., ( 2014 ) reported that heat stress negatively affects the

oxidative status of sheep along with the physiological responses and some of

these affects can be ameliorated through dietary antioxidants supplementation at

supranutritional concentrations.

Soliman et al.,(2012) administration of vitamin E plus Se to ewes was

accompanied with favourable signs on physiological reactions, blood

hematological profile and plasma metabolites . On the anther study Sobhanirad

et al, (2014) reported that supplementation of organic Zn either alone or in

combination with the inorganic Zn in the diet of growing lamb significantly

improved haematological and biochemical parameters .

The present study was aimed at elucidating the effects of zinc sulfate

administration and vitamin E + selenium (Se), as a antioxidant agents, on

thermoregulatory responses, hemato-biochemical parameters and reproductive

performance of Aberdeen Angus cows under New Valley conditions.

REVIEW OF

LITERATURE

Review of Literature

4

2 - REVIEW OF LITERATURE

In order to have a wide view on the objective of this study, the collected

literature will reviewed under the following items:-

2–1- Effects of using antioxidant on thermoregulatory responses:-

The rectal temperature and respiratory rates are recognized as important measures of physiological status (Lefcourt et al., 1986). Interestingly, the heart

rate was reduced by the antioxidant supplementation irrespective of thermal

treatment, which may be explained by the beneficial effects of Vitamin E on the

cardiac autonomic nervous system as has been reported in rats and humans (Behrens et al., 1986; Manzella et al., 2001; Fahim et al., 2013). There was

significant (p<0.05) increase in rectal temperature and respiratory rates in the

heat stress animal . The rectal temperature and respiratory rates in antioxidant

treatment groups (Vitamin C and E) were significantly (p<0.05) lowered. The

decreased rectal temperature and respiratory rates in treatment groups indicated

that supplementation of vitamin E with selenium and vitamin C ameliorated the heat stress in goats (Sivakumar et al., 2010) . Similar decrease in rectal

temperature and respiratory rates by vitamin E and C supplementation was

reported by Shenglins et al. (2003) in pigs and Kobeisy (1997) in goats under

heat stressed conditions. Previously, dietary antioxidant supplementation was

shown to reduce rectal temperature and respiration rate in goats under heat stress

where there was a small reduction (approximately –8%) in feed intake in

response to heat stress although not as great as in those not supplemented with

antioxidants (–15%; Sivakumar et al., 2010).

Chauhan et al. (2014) confirmed that supranutritional doses of Vitamin E

and selenium can reverse some of the negative effects of heat stress in sheep by

improving its oxidative status. Hence, we accepted the hypothesis that

supranutritional antioxidant supplementation can reverse some of the negative


5

effects of heat stress on oxidative status and improve some of the physiological

responses to heat stress in sheep.

Alhidary et al. (2012) reported that multiple injections of large doses of

selenium can be used to reduce or alleviate the consequences of thermal stress,

including the increased rectal temperature and BW loss, which has important

implications for the sheep industry . The 5 mg selenium treatment decreased

rectal temperature by 0.3°C (P = 0.02) and increased eosinophil count (P <0.05). There were no differences between treatments in Respiration rate (Alhidary et

al ., 2012). In other study Tahmasbi et al. (2012) reported that treated Holstein

cows with selenium-vitamin E were observed to be higher rectal temperature

and less respiratory rate than non-selenium-vitamin E group (p>0.05) during hot

weather. With no significant differences in rectal temperature and respiration rate. Chauhan et al. (2014) found that respiration rate increased in response to

heat stress (89.1 vs. 161 breaths per minute for antioxidant treatment (vitamin E

and selenium) and heat stressed conditions, respectively. Also, rectal

temperature increased in response to heat stress (39.45 vs. 40.03°C for

antioxidant treatment (vitamin E and selenium) and heat stressed conditions ,

respectively of lambs.

Daghash and Mousa (2002) showed a significant decrease ( P < 0.01) in

rectal temperature in T1 supplemented with 50 ppm Zn and T2 supplemented with 100 ppm Zn as compared to the control group. Also , Chirase et al . (

1991) reported that steers fed control diet had higher (P0.10) rectal temperature

than the steers fed Zn supplementation.

Khalifa et al. (2016) indicated that selenium treatment either as inorganic

or organic forms with or without vitamin E did not affect rectal temperature and

respiration rate of Holstein cows under Egyptian summer conditions. Also , El-

Shahat and Abdel-Monem (2011) showed that there was no significant

differences in rectal temperature and respiratory rates of Egyptian Baladi ewes


6

that treated with Vitamin E and selenium under subtropical conditions. On another hand Nayeri et al. (2014) reported that feeding complexed zinc had no

effects for rectal temperature of Holstein cows regard less of level .

2-2 Effects of using antioxidant on blood hematological parameters :-

2-2-1. White blood cells and its fractions :

It has been explained that the higher leucocytes and lymphocyte cell

counts due to selenium and vitamin E administration could be related to the

protection of cell membrane and intracellular organelles by the antioxidant

effects of selenium and vitamin E and thus increase their lifespan (Moeini and

Jalilian 2014). Supplementation of selenium alone or vitamin E plus selenium

had no effect on total count of leucocytes in buffalo calves (Shinde et al., 2009).

Also, selenium alone had no effect on neutrophil % in buffalo calves (El-Ayouty et al., 1996), while neutrophil phagocytosis was improved in vitamin E

plus selenium supplemented-heifers (Suwanpanya et al., 2007). In Awassi

rams, neutrophils percentages were found to be decreased with supplemental vitamin E alone vs. selenium plus vitamin E (Ammar et al., 2009) .The

synergistic effect of vitamin E and selenium to enhance lymphocyte function and its proliferative responses was detected with cattle (Pollock et al., 1994).

Injection of vitamin E alone enhanced lymphocyte populations in calves (Reddy et al., 1986). However, supplementation of selenium alone at different levels in

buffalo calves did not alter the lymphocyte % (El-Ayouty et al., 1996). So, it

appears that the usefulness of selenium to the immune function was increased by

giving vitamin E (Nockles 1988) and the combination of vitamin E and

selenium were more effective than selenium alone to improve immune system

(Fazaeli and Talebian 2009 ). The enhanced effect of vitamin E and selenium

to increase lymphocytes (%) agree with similar response in Ossimi sheep

(Soliman et al., 2001) , and buffaloes (Qureshi et al., 2001) . In addition, the

effect of vitamin E plus selenium increasing total count of leucocytes observed

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7

on Friesian heifers (Suwanpanya et al., 2007) adult buffaloes during late

gestation (Qureshi et al., 2001) growing Ossimi lambs (Soliman et al., 2001)

and dairy calves (Mohri et al., 2005) .

El-Ayouty et al.(1996) found that supplement of selenium alone to

suckling buffalo calves had no effect on neutrophil percentages. But, Suwanpanya et al. (2007) illustrated that neutrophil phagocytosis was improved

in vitamin E plus selenium supplemented heifers . Soliman et al. (2012) found a

significant increase in lymphocytes percentages either for vitamin E plus

selenium-injected ewes or their male lambs. The enhanced effect of injectable

vitamin E and selenium to increase lymphocytes (%) was previously reported on growing lambs (Soliman et al., 2001) and adult buffaloes (Qureshi et al.,

2001).

It has been noticed that lymphocyte proliferation was declined following

prolonged exposure of lambs to a diet deficient in both vitamin E and Se, while

their supplementation restored lymphocyte function within a week (Turner and

Finch 1990). The enhanced lymphocyte function and its proliferative responses

due to the interacted effect of vitamin E and selenium were also reported with

cattle (Pollok et al., 1994).

White blood cell and LYM differed significantly (P<0.05) between

control and Zn-supplemented groups as Zn supplementation increased these

parameters compared to control ( Southern and Baker 1983). King and

Fraker (1991) reported that reduction of the total number of WBC, especially

LYM, is the primary cause of depressed host defense capacity in Zn deficient adult mice . Moreover, Shinde et al. (2009) found that total count of leucocytes

was not affected by selenium alone or vitamin E plus selenium supplementation to buffalo calves. El-Amin et al. (2013) found that differences of WBCs were

not significantly affected by zinc supplementation (33mg zinc/kg) of goat Kids


8

in Sudan . Also, Cope et al. (2009) reported that no significant effect (P > 0.05)

on WBCs of zinc supplementation on dairy cow.

2-2-2. Red Blood Cells :

Qureshi et al. (2001) reported significantly (P<0.05) higher haemoglobin

concentration (Hb), red blood cell (RBC) count and hematocrit (HCT), while

mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and

mean corpuscular hemoglobin concentration (MCHC) remained unchanged in

buffaloes supplemented with vitamin E and selenium. In other study, Shetaewi et al. (1992) found that supplemented vitamin E at rate 100 mg/head/weekly

increased RBCs, HB and HCT in coarse-wool lambs. While in case of

supplementation of selenium alone to lambs, resulted to increase total RBCs count (Faixova et al., 2007) . However, El-Ayouty et al. (1996) found that

selenium alone had no marked effect on RBCs count, Hb and hematocrit in

buffalo calves. Likewise, supplementation of different forms of selenium did not

markedly influence the dynamics of blood parameters in non-pregnant, pregnant

and lactating ewes if the intake of vitamins and other essential microelements was adequate (Pisek et al., 2008) .

The significant increase in RBCs, blood Hb and HCT (%) due to injection of vitamin E plus selenium on Ossimi sheep (Soliman et al.,2001 and Soliman

et al., 2012) and on dairy calves (Mohri et al., 2005). In other study, selenium

supplementation at 0.3 mg Se/kg DM as sodium selenite had no effect on blood

Hb and HCT of goats (Kamdev et al., 2015). Also, supplementation of selenium

and vitamin E in the diet of buffalo calves had no significant effect on hematological parameters (Hb, HCT and RBCs) as reported by Shinde et al.

(2009) . The elevation in HCT % may be related to the antioxidant effect of

vitamin E and selenium that reduce the hemolysis of blood erythrocytes

(Makkawi et al., 2012).


9

Bremner et al. (1976) reported that higher level of Zn (420 mg/kg)

induced depression in haematological parameters such as blood Hb

concentration and HCT when compared with diets providing 48 mg/kg Zn in

sheep. In addition, dietary zinc supplementation (33mg zinc/kg) had no effect on

RBC, HCT and HB concentration of Goat Kids in Sudan had reported by El-

Amin et al. (2013) . Also , Southern and Baker (1983) did not find any

change in blood Hb or HCT of the male chicks supplemented with Zn at the rate of 50 and 100 mg/kg. Also Cope et al. (2009) reported no significant effect (P

> 0.05) on HGB, RBCs , HCT , MCV , MCH and MCHC of zinc

supplementation on dairy cow .

2-3 Effects of using antioxidant on blood biochemical parameters:-

2-3-1: Blood total protein and its fractions:

Soliman et al.(2012) reported a significant increase in total protein and

globulin for ewes and there lambs treated with vitamin E and selenium, with no

changes on albumin . Similar response of elevated serum total protein and

globulin were obtained in buffaloes supplemented with vitamin E and selenium

during the last stage of pregnancy (Helal et al., 2009). However, Shinde et al.,

(2009) found no effect on serum total protein and globulin for supplementation of selenium and vitamin E in the diet of buffalo calves. While, Rahmani et al.

(2015) treated early-lactation cows with vitamin E were significant increase on plasma albumen and plasma total protein. Also , Shokrollahi et al. (2013)

reported that serum levels of total protein, albumin, and globulins were

significantly (P<0.05) different with selenium and Vitamin E treatments on

Newborn Goat Kids.

Soliman (2015 b) injected two groups of growing lambs with two doses

from vitamin E and selenium the first 0.5 ml/head ( 3.57 IU Vitamin E + 0.03mg

selenium /heed/day ) and second 1.0 ml/heed ( 7.14 IU Vitamin E + 0.06mg


10

selenium /heed/day ) , respectively . The results indicated that lambs gave

second dose were higher (P<0.05) of serum total protein and globulin than those

gave first dose and control . while no significant differences on serum levels

albumin, among all groups. In Baladi sheep, supplement of vitamin E plus

selenium had increased serum globulins, with no significantly differences in

serum total protein and albumin (Hamam et al., 2007) . Also, Ziaei (2014)

showed that supplementation of diets with different levels of selenium and

vitamin E had no significant effect on total proteins and albumin. Moreover, Kumar et al. (2009) observed that supplementation of selenium at 0.15 ppm

level either from organic or inorganic source had no effect on the values of

serum total protein, albumin and globulin . In another study, Supplementation

of selenium to male buffalo calves had no significant effects on concentrations

of total protein, While, the level of globulin was significantly increased, leading to reduced levels of albumin ( Mudgal et al., 2008) . Moreover, Bagnicka et

al. (2017) reported no differences in serum total protein and albumins with

selenium in the organic vs. inorganic form of dairy cows . Also, Dagash et al.

(1993) observed that serum total protein and globulin concentrations did not

differ as a result of vitamin E injection . While, the serum albumin tended to be

higher in treated group than control one .

Mudgal et al. (2012) found that supplementation of Se, Cu, and Zn had

not any significant effect on total protein, albumin, globulin and other investigated blood biochemical parameters in the same animals. Also, Hassan et

al.(2016) revealed that there were no significant effects (P>0.05) in

concentrations of serum total protein, albumin, and globulin due to the Zn-

methionine or Zn-sulphate supplements for calves. Farghaly et al. (2017)

revealed that no significant differences (P>0.05) in serum total protein, albumin,

globulin, of lambs due to Zn-supplemented diet.While, Mousa and EL-Sheikh

(2004) indicated that zinc sulfate addition increased total protein and globulin


11

concentration , while it decreased albumin concentration in blood serum of

buffalo calves. Also , Daghash and Mousa (2002) reported a significant

increase of total protein and total globulin in buffalo calves after Zn

supplementation over 180 days with no significantly change in albumin. In another study, El-Amin et al. (2013) showed that dietary Zinc supplementation

(33 mg Zinc/kg) did not significantly (P>0.05) affected levels of serum total protein . Also, Shakweer et al. (2010) showed that addition of zinc sulfate or

zinc methionine had no significant effect on plasma total protein concentration .moreover Shakweer et al. (2006) found normal concentrations of total protein,

globulin and zinc with different levels of zinc methionine supplementation.

Zeedan et al. (2014) found a significant increase (P<0.05) in total protein,

albumin, and globulin of goats that supplemented with Biogen–Zinc .

2-3-2 : Total cholesterol :

Rahmani et al. (2015) treated early-lactation cows, beginning 5 weeks

postpartum with vitamin E were more effected on serum cholesterol concentration . Also , Nayyar et al. (2003) reported that supplementation of

vitamin E (500 IU) and vitamin E + selenium (500 IU vitamin E + 2 mg

selenium) per head per day to anestrous buffalo heifers caused a significant

(P<0.01) increase in the levels of cholesterol, as compared to unsuppleemnted anoestrous buffalo heifers. Moreover, Shetaewi et al. (1992) reported that

supplemented Vitamin E at dose 100 and 200mg increased cholesterol

concentration in serum of ewes and lambs. While, dietary supplementation of

selenium and vitamin E significantly decreased total plasma cholesterol concentrations in sheep (Gabryszuk et al., 2007) . Also, Soliman (2015 b)

observed that serum cholesterol concentration decreased (P<0.5) for lambs

received two doses from vitamin E and selenium vs. control .

Some authors reported no effect on the blood cholesterol levels

supplementation with selenium at a level of 0.1 and 8.54 ppm, respectively in


12

Friesian steers (Arthur et al., 1988) and in buffalo calves (Singh et al., 2002).

Also, Kumar et al. (2009) found that supplementation selenium to lambs at rate

of 0.15 mg Se/kg of diet through sodium selenite (inorganic selenium ) and

Jevsel-101 (organic selenium) , had no effect on serum total cholesterol and antioxidant status of the lambs. Sushma et al. (2015) reported that

supplementation of selenium in the form of sodium selenite (inorganic source) at

different levels in growing Nellore ram lambs had no effect on serum Cholesterol . Furthermore, Bagnicka et al. (2017) reported that there were no

differences in blood serum cholesterol with selenium in the organic vs. inorganic

form of dairy cows.

Farghaly et al. (2017) reported that serum cholesterol was higher (p <

0.05) in Zn treated groups than in control one. Moreover, Zeedan et al. (2014)

reported that Biogen–Zinc improved total cholesterol in blood plasma within late pregnancy and lactation period of goat . While, Hassan et al. (2016)

reported that average value of cholesterol for calves fed diets supplemented

with Zn-methionine or Zn-sulphate were significantly lower than those fed control (196.96 and 209.61 vs. 230.47 respectively) . Also, Mirosław et al.

(2007) reported that enrichment of diet with Se, Zn and vitamin E (0.42, 68 and

60 mg /lamb/day, respectively) significantly decreased the cholesterol content of

loin muscle and the level of total cholesterol of blood plasma. In other hand Shinde et al. ( 2013) found that cholesterol levels of lambs fed Cu- and Zn-met

or Cu- and Zn-sulphate were comparable (p=0.05) . Also , El-Amin et al.

(2013) showed that dietary Zinc supplementation (33 mg zinc/kg) did not

significantly (P>0.05) affected levels on serum cholesterol of goat kids .

2-3-3 : Glucose :

Nayyar et al. (2003) observed significantly (P<0.01) higher level of

blood glucose in anoestrous buffalo heifers supplemented with vitamin E + selenium as compared to control group. Also, Alhidary et al. (2015) found


13

an increase in the serum concentrations of glucose in sheep fed diets supplemented with selenium and vitamin E. However, Singh et al. (2002)

observed low blood glucose concentration in buffalo calves fed wheat straw containing high selenium (8.54 ppm). In contrast, Shinde et al. (2009) reported

that the supplementation of selenium, vitamin E or both had no effect on the

serum glucose level of buffalo calves . Also, Sushma et al. (2015) reported that

supplementation of selenium in the form of sodium selenite (inorganic source)

at different levels in growing Nellore ram lambs had no effect on blood glucose

Soliman et al. (2001), Soliman et al. (2012) and Soliman (2015b)

reported that injection / supplementation of vitamin E and selenium had no significant effect on serum glucose concentrations in sheep . Also Kamdev et

al. (2015) found that supplementation of selenium alone had no significant

effect on serum glucose concentrations in goats. In addition , Shinde et al.

(2008) observed that supplementation of vitamin E (300 IU) and/or selenium

(0.3 ppm) in the diet had no effect on serum glucose levels in buffalo calves. The same result included by Amirifard et al. ( 2016 ) also found that plasma

concentrations of glucose were not affected in dairy cows supplemented with vitamin E. Also, Bagnicka et al. (2017) reported that there were no differences

in blood serum glucose, with selenium in the organic vs. inorganic form of dairy

cows.

Dietary zinc supplementation had significant (p=0.05) increase on blood serum glucose of goat kids in Sudan (El-Amin et al., 2013). Furthermore,

Zeedan et al. (2014) reported that serum glucose were significantly increased

(P<0.05) with supplementing Biogen- Zinc of Damascus goats. Also, El Hendy et al. (2001) showed that plasma glucose concentration in two test groups which

were fed with insufficient zinc diet was significantly lower than controls. While, Cortinhas et al. (2012) found no effects of organic and inorganic sources of

zinc, copper and selenium in diets for dairy cows on plasma concentrations of


14

glucose . Also, Kinal et al. (2007) did not observe any effect of

supplementation of organic Zn, Cu or Mn from six weeks pre partum to three

months of lactation on concentrations of glucose for dairy cows . Also, Farghaly et al. (2017) revealed that no significant differences in serum glucose

of lambs due to Zn-supplemented diet . Also , Puchala et al. (1999) detected

that no effect of dietary Zn-met supplementation in Angora goats on plasma

glucose concentrations.

2-3-4 : Liver function (ALT and AST enzymes )

Previous studies have demonstrated significant associations between

selenium plus vitamin E and blood concentrations of alkaline aminotransferase (ALT) , asparagine aminotransferase (AST) in sheep (Cronje et al., 2006 ;

Surai, 2006; Braun et al., 2010) . Regarding selenium supplementation, Singh

et al. (2002) observed that buffalo calves fed high selenium (8.54 ppm) had

higher activity of plasma ALT and AST . Furthermore, Rahmani et al. (2015)

showed that treated early-lactation cows with Vitamin E had no effect on AST, however more effected on ALT . Moreover, Bagnicka et al. (2017) reported

that There were no differences in (ALT) and (AST) with selenium in the

organic vs. inorganic form in dairy cows. Also, El-Sharwy et al. (2017)

reported that AST and ALT activities in blood serum of Egyptian buffalo

bulls were not significantly different between the control group and selenium treated groups . Also, Metery et al. (1999) reported no significant differences in

AST and ALT activities in buffalo calves in respect to selenium treatments. In

addition, Soliman (2015 a) found no significant differences in serum

concentration of AST activity due to treatments Vitamin A , Vitamin E and

Vitamin A+E vs. control . The same results obtained by Kassab and

Mohammed (2014) and Soliman (2015 b) thy reported that Serum AST and

ALT concentrations not changed due to vitamin E plus selenium treatment .


15

Hassan et al. (2016) reported that the average value of ALT in blood

serum of calves fed diet supplemented with Zn-methionine was higher (P<0.05)

than those fed diet supplemented with Zn-sulphate (30.77 vs. 25 Unit/l).

However, no significant differences were observed among groups for AST. Also, Azizzadeh et al. (2005) showed that significantly higher AST activity in

Holstein calves supplemented with 50 ppm Zn . Similarly , Daghash and

Mousa (1999) observed higher ALT and AST activity in buffalo calves supplemented with 50 or 100 ppm Zn . While, Shinde et al. (2013) found that

ALT activities significantly (p<0.05) declined in Cu- and Zn-met group than

Cu- and Zn sulphate indicating an effective role of chelated minerals in

improving liver function .

Valle et al. (2015) reported that AST activities were not affected by

organic and inorganic sources of minerals in diets for dairy cows . Also, no

significant changes (P>0.05) in AST and ALT activities were observed due to

Zn level, source or duration of feeding in crossbred calves ( Mandal et al.,

2008) . On another study in ewes, Hassan et al. (2011) reported that inorganic

or organic zinc did not cause any effect on AST and ALT activities .

2-3-5 : Kidney function ( Urea and creatinine )

Previous studies have suggested that the increase in creatinine

concentration during hot environmental conditions could be attributed to an

increase in muscle activity in the respiratory system and thoracic cavity caused

by an increased respiratory rate and increased oxygen consumption as a part of the thermoregulatory processes to maintain homestasis (Mendel et al., 2005;

Brosnan and Brosnan, 2010).

Reddy et al. (1987) did not notice any effect on serum urea and

creatinine values of calves, supplemented with graded levels of vitamin E.

While, An increase in serum creatinine concentration was associated with


16

selenium and vitamin E supplementation ( Aiello, 1998) . In other study, Mudgal et al. (2008) reported that supplementation of selenium to buffalo

calves had no effect on their serum urea and creatinine levels. Also , Kassab

and Mohammed, (2014) reported that injection of vitamin E plus selenium to

ewes for 4 weeks prior mating and 4 weeks after mating revealed no significant

effect in urea and creatinine concentration . Also, El-Shahat and Abdel

Monem, (2011) reported that serum urea concentration among different

supplement of Vitamin E and /or selenium were not significant different of

Egyptian Baladi Ewes

Fagari-Nobijari et al. (2012) reported that urea nitrogen concentration

were higher in animals fed ZnSO4. While, Zeedan et al. (2014) reported that

Biogen–Zinc caused asignificant decrease in plasma urea and creatinine concentration for dairy Damascus goats. Also, Shakweer et al. (2010) showed

that urea concentration in blood plasma was decreased either with zinc sulfate

or zinc methionine . Mousa and EL-Sheikh, (2004) indicated that zinc sulfate

addition decreased urea concentration in blood serum of buffalo calves. In addition, Hassan et al. (2011) reported that organic Zn (Zn-Met) caused a

significant decline (P < 0.05) in urea compared to the inorganic Zn . ut

inorganic or or-ganic zinc did not cause any effect on the serum creatinine

concentration.

Sobhanirad and Naserian, (2012) found no differences between organic

and inorganic zinc sources treatments in urea concentrations of blood serum in

Holstein dairy cows. Also, Cortinhas et al. (2012) reported that organic or

inorganic sources of Zn, Cu, and Se did not affect on plasma concentrations of urea for Holstein cows . Furthermore , Kinal et al. (2007) did not observe effect

of supplementation of organic Zn, Cu or Mn from six weeks pre partum to three

months of lactation on concentrations of urea for dairy cows. While,

Sobhanirad et al. (2014) found insignificant differences in urea concentration


17

in lambs treated with different source and level of zinc. In addition, Valle et al.

(2015) reported no significant differences on urea concentration for cows by the mineral source in the diet. Also, Huerta et al. (2002) indicated that serum urea

nitrogen did not differ when feedlot heifers were supplemented with 200 ppm

Zn.

2-3-6 : Total antioxidant capacity (TAC)

Vitamin E and selenium (Se) play complementary roles as antioxidants.

Glutathione peroxidase (GSH-Px) is an enzyme involved in detoxification of

hydrogen peroxide and lipid hydroperoxides. Selenium as an essential

component of GSH-Px, acts to destroy peroxides before they attach cell

membranes, while vitamin E acts within the membrane to prevent the formation of fatty acid hydro peroxides (Milad et al., 2001) . Selenium and vitamin E

increases plasma concentrations of total antioxidant capacity and Selenium in dairy cattle (Calamari et al., 2011) and goats (Katamoto et al., 1998) .

In the antioxidant system Zn is a component of Cu–Zn SOD. Zinc also

induces synthesis of metallothionein, a metal binding protein that may scavenge

hydroxide radicals (Prasad et al., 2004). The role of Zn as an antioxidant (i.e.,

superoxide dismutase) and the importance of Zn to cell proliferation are the two

most direct connections between Zn and immune function ( Prasad , 2008 ) .

Khatti et al. (2017) reported that Treated cows with vitamin E and

selenium showed increased (P < 0.05) total antioxidant capacity (TAC) at -1, 0 and 3 to 8 weeks postpartum. Furthermore, Gong et al. (2014) reported that

TAC was higher (P<0.05) with Selenium yeast supplementation (organic source) than with selenite (non organic source) . Miller et al. (1993) also reported that

feeding 0 and 1,000 IU vitamin E during dry period to cows, led to significant

increase in the plasma total antioxidant activity at parturition. In addition,

Soliman, (2015a) reported a significant (P <0.05) increase in TAC for lambs


18

received vitamin A , vitamin E and vitamin A+E treatments vs. control. Also,

Dietary supplementation with 0.8 mg/kg seleniumand 150 mg/kg vitamin E for 50 d increased (P<0.05) plasma TAC in sheep (Alhidary et al., 2015) . In a

study on Boer goats, adding vitamin E (80 IU/ kid/d) can increase serum TAC (Hong et al., 2010). Also, Yu et al. (2007) reported that selenium

supplementation increased (P<0.01) the activity of TAC in erythrocyte

membrane in lambs.

The activity of antioxidant enzymes was significantly decreased by

supplementation of vitamin E and zinc indicating improvement in the

antioxidant activity and decrease oxidative stress to cows ( Maurya et al.,

2014). The inorganic Zn (ZnSO4·7H2O) caused a significant decline (P < 0.05)

in the antioxidants activity compared to the both organic Zn (Zn.Met) (Hassan et al., 2011). While, Rashidi et al. (2011) indicate that supplementation with Zn,

vitamin E or both of serum TAC did not show significant(P<0.001) changes in

any of the studied groups. Moreover, Farghaly et al. (2017) reported that serum

concentration of TAC were not affected by zinc supplementation. Also, Aliarabi et al. (2015) reported that TAC did not differ significantly with Zn

supplementation.

2-4 Effects of using antioxidant on reproductive performance:-

Studies on the relation of vitamin E supplementation to reproductive

parameters have generally produced contrasting results. Most studies

administered about 1000 IU of vitamin E and positive effects were found when selenium was also supplemented or adequate (Lacetera et al., 1996). Successful

attempts to improve the reproductive and productive efficiency by providing vitamin E selenium in exotic cows (Harrison et al., 1984), Sahiwal cows

(Sattar et al., 2003b&c), Egyptian buffaloes (Youssef et al., 1985) and Nili-

Ravi buffaloes (Qureshi et al., 1997; Sattar et al., 2003a) during late gestation

have been reported previously. Furthermore, vitamin E-selenium treatments


19

during the dry period are recommended for reducing postpartum reproductive

disorders in cows (Cortese, 1988).

Sharma et al. (2017) concluded that the strategic nutrients (Soya-DOC-

300 g, sodium dihydrogen orthophosphate dehydrate (NaH2PO4 .2H2O) - 11 g ,

zinc oxide-250 mg, copper sulphate- 169 mg , vitamin A (5 lakh IU/g) -210 mg ,

vitamin E 50% IU/100 g; vitamin E 50% IU/100 g) - 1225 mg per head/day in

gelatine capsules) supplementation in the ration of advanced pregnant crossbred

cows prevented reproductive problems and metabolic disorders . Trace minerals

are important for reproductive performance in livestock because their

supplementation improves reproduction (Chester-Jones et al., 2013) and also

via contributing to the normal health of reproductive organs and reproductive cycles ( Karkoodi et al., 2012 ) .

Earlier Manspeaker et al. (1987) reported the importance of trace

minerals in reproduction in cattle. Rabiee et al. (2010) reported higher

conception rates with organic trace elements in cattle. Trace minerals are important for reproductive performance in livestock (Kumar et al., 2011)

because their supplementation improves reproduction (Grace and Knowles,

2012). Boland, ( 2003) showed that the ovarian activity of ruminants is

influenced by mineral deficiency. Trace elements are also involved in synthesis

of hormones that are important for reproduction. Their deficiency affects both steroid (Boland, 2003) and thyroid (Abdollahi et al, 2013) hormone production.

2-4-1 Ovarian activity :-

A clinical deficiency in selenium can lead to many reproductive disorders

such as retained placentas, infertility, cystic ovaries, metritis, delayed

conception, and erratic, weak or silent heat periods leading to poor fertilization

(Corah and Ives, 1991) . During early fetal growth, selenium functions as a reducta to help protect against mortality from oxidative damage (Hostetler et


20

al., 2003) . When injected with supplemental selenium and vitamin E , beef

cows produced more fertilized ova when super ovulated than control animals that were not supplemented (Kappel et al., 1984). However, when cattle were

selenium adequate (210 – 1200 ng Se /mL in blood) and given supplemental

selenium and vitamin E via an injection there was no enhancement in

reproductive performance (Hidiroglou, 1987).

Feeding organic trace elements increased conception rates in cattle,

reduced days open (13.5 less days open) and number of services per conception (0.27 less services per conception) (Rabiee et al., 2010) and increased ovarian

activity in dairy cows (Boland, 2003) . Zinc and copper play an important role

in regulating progesterone production by luteal cells via involvement of superoxide dismutase (Sales et al., 2011) . Inadequate serum levels of zinc and

copper may induce decreased follicular growth and fertility, abnormal estrus,

anestrus, and abortions (O'Donoghue and Boland, 2002) . Generally, zinc is

known to be essential for proper sexual maturity (development of secondary

sexual characteristics), reproductive capacity (development of gonadal cells) in

males and all reproductive events (estrus, pregnancy and lactation), more

specific with onset of estrus in female. Among these decreased fertility and

abnormal reproductive events are of prime importance in females (Kumar,

2003).

Shorter days to first service have been attributed to combined effect of

mineral and vitamin supplementation because their positive effect on steroid

synthesis, release, follicular growth and symptoms of ovulatory oestrus (Srivastava, 2008). Moreover, Khan et al. (2015) reported that groups

supplemented with vitamin E and minerals showed better reproductive

performance than control group. Also, Supplementation of vitamin E and

selenium to Murrah buffaloes during prepartum period has been shown to cause

early initiation of postpartum ovarian activity and early exhibition of first


21

postpartum heat (Mavi et al., 2006). Also, Kose et al. (2013) reported that

injected ewes with vitamin E and selenium before estrus didn‘thave a positive

effect on fertility in synchronized ewes in anestrus season .

2-4-2 Conception rate :-

Many studies have reported a significant difference in conception rates

between treatment groups with selenium or selenium plus Vit E supplementation (Allan et al., 1993 and Castro et al., 2009). Several studies

have shown that administration Selenium, vitamin E or vitamin E and

Selenium in combination resulted in higher pregnancy rates in cows and heifers (Richardson et al., 2008 and Zanella et al., 2010) . Also, Qureshi et al.

(2010) showed that vitamin E and selenium administration significantly

(P<0.01) improved the conception rate and estrus rate . Moreover ,

supplementation of vitamin E and selenium to Murrah buffaloes during prepartum period to cause a significant increase in conception rate ( Mavi et al.,

2006).

Macpherson et al. (1987) reported that significant improvement in

conception rate of Friesian heifers by giving a single subcutaneous injection of

barium selenite. Also, Nayyar et al. (2002) reported a significant increase in

conception rate by supplementing 14 mg of selenium per week in buffalo

heifers. Moreover, Hemingway , (2003) reported an increase in conception rate

by providing a sustained release multi-trace element rumen bolus in beef cattle.

He attributed this increase in conception rate as the effect of selenium on

establishment of ova and sperm transport. In another study, Yavuzerand and

Bengisu, (2014) showed no effect of organic selenium supplementation on

pregnancy rates of Awassi ewes.

In beef cows grazing native range with access to a free choice

supplementation of Cu, Mn, and Zn, cows receiving a trace mineral injection pre


22

and post-partum exhibited higher conception rates to fixed timed artificial insemination over a control group (Mundell et al., 2012). In beef heifers that

were provided a free choice supplementation of Cu, Mn, and Zn, had a greater

conception rate to timed embryo transfer when compared to control heifers (Sales et al., 2011). In another study, Kundu et al. (2014) reported that

supplementation of 50 to 100 ppm zinc oxide to the basal diet of Teressa goat

significantly improved different productive and reproductive parameters , such

as significantly increased the incidence of oestrus (33%), pregnancy rate (12%),

kidding rate (5%) and resulted in shorter onset (8 days) of oestrus as compared

to the control. In addition, feeding organic trace elements increased conception

rates in cattle, (Rabiee et al., 2010) Furthermore, Ahola et al. (2004) reported

that Supplementation with Cu, Zn, or Mn improved pregnancy rate to artificial insemination compared with un supplemented cows. Also, Stanton et al. (2000)

reported that cows receiving organic trace minerals exhibited higher pregnancy

rates to artificial insemination than those receiving inorganic trace minerals . While, Baker et al. (2002) found no differences in overall conception rates due

to Cu, Zn, or Mn supplementation with cows. Olson et al. (1999) reported no

difference in reproductive performance in cows supplemented with organic vs.

inorganic forms of trace minerals. However, supplemented cows had lower

pregnancy rates compared with controls . Reduction in pregnancy rates can occur as a subclinical effect of marginal trace mineral status (Hostetler et al.,

2003) . thay reported that deficiencies in Cu, Se, Mn, and Zn have been linked to

abnormal estrus cycles, impaired ovulation and decreased conception rates (Hostetler et al., 2003 and Underwood, 1981) .

2-4-3 Service per conception:-

Pontes et al. (2015) reported that addition of Vitamin E to cows

improved pregnancy per insemination due to decreased pregnancy loss from days 31 to 62 of gestation (12.5 vs. 20.5%). Also, Jin Yu et al. (2015) indicate


23

that nano-selenium or selenium yeast Supplemented to cows diets can

significantly improve the reproductive performance and immune function. In addition, Bayril et al. (2015) showed that supplemented selenium and vitamin

E decreased the number of services per conception and the services period of

dairy cattle. Furthermore, Feeding organic trace elements reduced days open

(13.5 less days open) and number of services per conception (0.27 less services per conception) (Rabiee et al., 2010) . The services per conception were also

reduced from 2.50 (control) to 2.17 in supplementing Vitamin E groups. (Panda et al., 2006) . Moreover, Supplementation of vitamin E and selenium to

Murrah buffaloes during prepartum period has been shown to cause a significant

decrease in service per conception (Mavi et al., 2006) .

Some authors reported a positive response to the complex organic-

inorganic microelements compared to inorganic sources on first-service conception rates: 27.0 versus 21.0% (DeFrain et al., 2009) , and27.4 versus

18.4% (Ballantine et al., 2002). Campbell and Miller, (1998) observed

reduced days to first estrus and a tendency for reduced days to first service in

cows fed 74 mg/kg of supplemental zinc [50:50 blend of zinc sulfate and zinc

amino acid complex] during the last 6 week of gestation. In other study, Nayeri et al. (2014) reported that feeding Complexed zinc decreased the number of

services per conception in Holstein cows regardless of level .

Ballantine et al . (2002) reported a decrease in days open and a tendency

for an increase in cows pregnant at 150 DIM and first service conception risk in

cows supplemented with Zn, Mn, Cu, and Co complexes. Interestingly, Nocek et

al. (2006) continued treatments into a second lactation with the same cows,

where they noted that cows cycled sooner after calving, had higher first-service

conception risk, and had more cows pregnant by 150 DIM. Another trial also

reported a tendency for increased percentage of cows pregnant at 150 DIM in

cows fed a mixture of sulfates and Zn, Mn, Cu, and Co complexes compared

javascript:void(0);


24

with similar levels of those minerals provided as sulfates ( DeFrain et al.,

2009).

2-4-4 Estrogen and progesterone hormones:-

It is known that vitamin E and Selenium are an antioxidant, and could

play a role in the antioxidant system in the corpus luteum (Kamada and

Hodate, 1998). The concentration of progesterone was higher when vitamin

supplemented than control group (5.31 vs. 3.73 ng /ml) respectively. However

this concentration was lower after estrus (4.43 vs. 2.25 ng /ml respectively) for

supplemented and control group (Akomas and Ezekwe, 2013). Moreover, Kamada et al. (2014) reported that selenium supplementation increased

(p<0.05) plasma progesterone concentration in cows at the 29-39 weeks of pregnancy from 4.98 to 6.86 ng /ml. similarly, Makkawi et al. (2014) suggest

that selenium increased progesterone on the establishment of pregnancy especially in cows of lower fertility, is warranted. Forthermore, Ali et al. (2017)

reported that significant increase on estrogen in dairy cows that injected with vitamin E and selenium compered to control . Also, Ahmed et al. (2000)

observed that progesterone concentration was higher in females treated with

vitamin E than untreated. Similar results observed when New Zealand rabbits

treated wish vitamin E (50 mg /kg diets ) during the gestation period, which

increase progesterone concentration as compared with control group (Hassanein et al., 1995).

Copper and zinc play an important role in regulating progesterone production by luteal cells via involvement of superoxide dismutase Sales et al.

(2011) . Zinc is involved in the reorganization of ovarian follicles which are the

source of progesterone. This occurs through the involvement of

metalloproteinase-2 (MMP-2) enzyme, which is a member of zinc

endopeptidase family Gottsch et al. (2000). Positive correlation was reported

between serum progesterone level and copper-zinc in cows by Yildiz and Akar,


25

(2001). The ability of heifers to reach sexual maturity by the start of the

breeding season is dependent on adequate concentration of hormones involved in development of the reproductive tract (Andersen et al., 1991; Day and

Andersen, 1998). It has been stated that Cu, Zn, and Mn influence reproduction in mammals, possibly through hormone synthesis (Hambidge et al., 1986 and

Davis and Mertz, 1987).

MATERIALS AND

METHODS

Materials and Methods

26

3 - MATERIAL AND METHODS

This experiment was carried out at the Animal Production Experimental

farm of Faculty of Agriculture, New Valley University, cooperation with

Animal production Department in Faculty of Agriculture, Assiut University. The

study was started from July to September 2016 in the new valley governorate

during summer season. New Valley governorate located in Upper Egypt also, in

western desert between 25º ;42º & 30º;47º E longitude , 22º;30 & 29º;30º N

latitude and lies 77.8m a latitude above the sea level . The climate of this area is

arid and dry, essentially that of the desert. Rainfall is almost negligible and the

ambient temperature ranges from 46º C during summer days to 8º C in chilly

winter nights. Data were collected from the Aberdeen Angus cattle flock in

college . This herd was imported from the state of Uruguay through the

Egyptian Ministry of agriculture . The aim of the current study was to

determine the interaction effect of vitamin E , selenium and zinc sulphate on

reproductive performance, blood hemato-biochemical parameter and resisting

thermal stress of Aberdeen angus cows.

Animal and experimental design

A total number of sixteen healthy Aberdeen Angus cows about 3-4 years

of age and an average body weight of 460-520 kg were used in this study . The

animals were divided randomly in to four equal (4 animals each groups) .

Control ( C ) , vitamin E and selenium (T1) , zinc sulphate (T2) , and vitamin E

and selenium plus zinc sulphate (T3) ). All cows were fed on basal diet which

was formulated according to NRC, (2000) for Beef cattle.

The basal diet consists of 40% wheat straw and 60% concentrate mixture.

Cows in first group (C ) were fed a basal diet without any supplementation

whereas , cows in second group (T1) was intramuscular injected every two

weeks for 90 days with 15 ml viteselen. Each 1ml of viteselen contains vitamin


27

E, 150 ml acetate and 1.67mg sodium selenite . The third group (T2) fed daily

with zinc sulphate 200mg/heed While, the fourth group (T3) was fed daily with

zinc sulphate 200mg/head and injected every two weeks for 90 days with 15 ml

viteselen (vitamin E and selenium).

Table (1): Composition of concentrate feed mixture and treatments .

Items Treatment

Control T1 T2 T3

Yellow Corn% 55 55 55 55

Wheat bran% 21.5 21.5 21.5 21.5

Soyabean meal% 20 20 20 20

Limestone % 1.5 1.5 1.5 1.5

Dicalcium phosphate% 0.5 0.5 0.5 0.5

Yeast % 0.2 0.2 0.2 0.2

Bicarbonate % 0.3 0.3 0.3 0.3

Sodium chloride % 1 1 1 1

Vitamin E and selenium (ml / head) 15/15day 15/15day

Zinc sulphate (mg / head) 200/day 200/day

C: Control, T1:injected with vitamin E and selenium , T2: supplementation with zinc sulphate, T3: injected

with vitamin E and selenium and supplementation with zinc sulphate

Table (2): Chemical composition of concentrate mixture and wheat straw.

Item DM OM CP CF Fat Ash NFE

Concentrate mixture 88.76 93.79 15.76 14.12 2.39 6.21 61.52

Wheat straw 90.35 89.05 1.79 38.71 1.12 10.95 47.43

DM: Dry Matter, CP: Crude Protein, CF: Crude Fiber, NFE: Nitrogen Free Extract


28

The experimental design was divided in to two parts :-

3-1. The first part :-

It was carried out to investigate the effect of different treatments on

resisting thermal stress of Aberdeen Angus cows.

3-1-1. Meteorological parameters :

Air temperature and relative humidity were recorded during the

experimental days using Temperature /Humidity thermometer at 2 pm and 10

pm .

Temperature Humidity Index (THI) was calculated according to Mader et al. (2006) as following THI = (0.8 × Ta) + [(RH/100) × (Ta – 14.4)] + 46.4

Where; Ta °C is the ambient temperature (°C), and RH is the relative humidity

(RH %) /100.

3-1-2. Thermal responses:

3-1-2-1. Respiration rate

Respiration rate (RR) was determined per 21 day by counting the flank

movements for a minute (breath / min.)

3-1-2-2. Pulse rate

Pulse rate ( PR ) was determined per 21 day, the pulse rate was counted

before measuring the body temperature .

3-1-2-3. Rectal temperature

Rectal temperature (RT) was measured per 21 day using a standard

clinical thermometer inserted gently into the rectum for one minute.

3-1-2-4. Skin temperature and Hair temperature

Skin temperature and hair temperature were recorded per 21 day using

Infrared Thermometer (-27º to 230º F - 33º to 110º C range )


29

3-1-2-5. Ear temperature

Ear temperature inside and outside the ear were recorded per 21 day

using Infrared Thermometer (-27º to 230º F - 33º to 110º C range )

3-1-3 Blood sampling and analytical methods

3-1-3-1 Collection of blood

About 12 ml blood was collected from each cows through jugular

venipuncture in the morning (before watering and feeding) at day 0, 30, 60 and

90 of the experimental period. Out of 12 ml, 10 ml blood was collected into

clean and dry test tube and kept in slanting position for 45 min for the separation

of serum for blood biochemical parameters . Remaining 2 ml was taken in

another clean and dry ependroph tube (2 ml) containing anticoagulant (EDTA)

for the haematological studies. About 10 ml blood was collected at day 0, 3, 6,

9, 12, 15, 18 and 0 of the estrus cycle into clean and dry test tube and kept in

slanting position for 45 min for the separation of serum for progesterone and

estrogen hormone.

The blood samples were centrifuged at 3000 rpm for 10 min at 4oC and

serum was separated. The serum was collected in plastic vials and kept at - 20oC

until further analysis.

3-1-3-2 Hematological parameters

Anti-coagulated blood was analyzed shortly for the number of white

blood cell (WBC) count, Lymphocytes (LYM), Granulocytes (GRAN) ,

Monocytes (MID) , red blood cell (RBC) count, hemoglobin concentration (Hb),

hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular

hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and

red Blood Cell Distribution Width (RDW), were calculated by an automatic

hematology cell counter (sysmex).


30

3-1-3-3 Serum biochemical parameters

Stored serum samples were analyzed for total protein (TP), albumin

(ALB), glucose (GLU), urea nitrogen (UN), creatinine, total cholesterol (TC),

alanine aminotransferase ( ALT) , and aspartate aminotransferase (AST) were

determined colorimetrically using Bio-diagnostic product kits (Egypt). Serum

globulin concentrations were calculated by difference between total protein and

albumin concentrations. Total antioxidant capacity (TAC) were analyzed

colorimetrically by jenway spectrophotometer, using Bio-diagnostic kits (Bio-

diagnostic Company, Egypt). Progesterone levels were determined using

ELISA. Kits were provided by BioChek Inc. (Foster City, CA, USA). The range

of the standards used was 0.5 to 50.0 ng/mL. Assay sensitivity was 0.22 ng ⁄ ml,

and a 50% effective dose (ED50) was 1.4 ng ⁄ ml. Intra- and interassay

coefficients of variation were 5.2 and 10.4%, respectively. Estadiol hormone

was determined using ELISA. Kits were provided by Diagnostic System

Laboratory Co. (DSL, Catalogue No. 3900, USA).

The analyses were performed at, Animal Production Department, Faculty

of Agriculture, New Valley Branch, Assiut University.

3-2. The second part :-

It was carried out to investigate the effect of experimental treatments on

reproductive performance of cyclic Aberdeen Angus cows.

3-2-1. Estrous synchronization using prostaglandins

The hormone, prostaglandin (PG), was the first available for estrous

synchronization. Prostaglandin systems work equally well for cows as well as

heifers. At least three products are commercially available, all by prescription

and all seem to work well. Cows must be cycling for PG to show a significant


31

effect. Prostaglandins are not generally recommended for timed insemination

without the use of other hormones.

This system, (A) is used to bring more cows in heat during the Artificial

insemination (AI) period (90%). Heat detection must be practiced. If extended

heat detection is not a concern but drug costs are a problem, then animals can be

heat checked and bred after the first injection of PG (B). Animals not showing

heat following the first injection would then be administered a second

prostaglandins injection 11-14 days later and then bred.

Fig. (1): Estrous Synchronization Using Prostaglandins

3.2.2 Trans-rectal ultrasonographic examination

3.2.2.1. follicular development and corpus luteum:

The ovarian structures (follicles and corpora lutea) were investigated

through transrectal ultrasonography by a real-time B-mode mobile US unit with

a 5 ⁄ 7.5-MHz linear array transducer (ECM, Noveko International, Inc.,

Angouleˆme, France). The US equipment was supplied with image freeze and

electronic calliper functions for taking measurements.

The animals were examined by the same operator every 3 days from the

day of estrus (day 0) till the next estrus at days 0, 3, 6, 9, 12, 15 18 and 0.

Oestrus was detected in combination with ultrasonography and by a teaser bull

with cows during the experimental period. During the trans-rectal

ultrasonography procedure, the operator removed the feces from the rectum,

introduced the transducer, and avoided grasping the uterine horns. The

transducer was cleaned between animals. During the transrectal ultrasonography


32

procedure, the operator removed the feces from the rectum, introduced the

transducer, and avoided grasping the uterine horns. The transducer was cleaned

between animals

Follicles were defined as non-echogenic rounded structures with a clear

demarcation between the follicular wall and antrum. A corpus luteum (CL) was

defined as a grainy echogenic structure that had a well-defined border with the

less echogenic ovarian stroma, and in some corpora lutea, there was a non-

echodense lacuna (Sheldon et al. 2002). The maximum diameter of each

structure was measured using the electronic calliper. When the image of the

structure being scanned was not circular, the diameter was estimated by

averaging two dimensions at 90º (Sheldon and Dobson 2000). The animal

considered ovulated when an ovulatory follicle (10–23 mm) detected in the

preceding examination disappeared with the formation of ovulation depression

and assured by the formation of a CL in the subsequent examination.

Ovarian follicles were counted and classified according to their diameter

into small (<5 mm), medium (5–8 mm), large (9-12) and ovulatory follicles

(>12 mm). The diameter of corpus luteum and size of largest follicle (mm) were

recorded.

Cows showed estrus have been mated with proven fertile bull at the

appropriate time (12 hrs after the appearance of estrus signs). for two successive

estrous cycles till pregnancy occur

3.2.2.2. Pregnancy diagnosis:

Pregnancy was estimated by trans-rectal ultrsonography between 35 and

65 days post mating. The diameter of corpus luteum gravidatis, size of embryo

(mm) and size of amniotic vesicle (mm) were recorded.


33

3-2-3 Reproductive parameters of cows:

x Percentage of estrus (Oestrus response) : Calculated as (estrus cows /

Synchronized cows)

x Number of services per conception: Calculated as number of all services

to the cows until the successfully conceiving.

x first services conception

x second services conception

x Conception rate: Calculated as (pregnant cows / cows mated) x 100 .

3-3. Statistical Analysis :-

The data were analyzed using a completely randomized design with the

GLM procedure of the statistical program SAS/STAT 9.1 (SAS, 2004).

The differences among treatments were tested using Duncan's Multiple

Range Test (Duncan, 1955).

The model used was Yij = U + Aj + Eij

Yij = Observation traits

U = Overall mean

Aj = Experimental treatment

Eij = Random error

Pregnancy rate values statically tested using Chi square test.

Correlation values were calculated between ovarian follicular and luteal

development and hormone profile.

RESULTS AND

DISCUSSION

Results and Discussion

34

4 - RESULTS AND DISCUSSION

4-1- Ambient temperature, relative humidity and temperature-humidity

index (THI) during experimental period.

The values of ambient temperature, percent of relative humidity (RH) and

temperature humidity index (THI) during the experimental period were shown in

Table (3) and Figure (2).

Table (3): Means of ambient temperature (°C), relative humidity (%) and temperature

humidity index (THI) during the experimental period .

Items 2pm 10pm

0 day 21 day 42 day 63 day 84 day

Air temperature ( Taº C )

2 pm 40.3 45 43.6 38.9 38.3 10 pm 29.3 28.5 30.3 25.2 26.4

Relative humidity (RH%)

2pm 10 10 10 10 10 10 pm 25 21 22 23 22

(THI)1Temperature humidity index

2 pm 81.23 85.46 84.20 79.97 79.43 10 pm 73.57 72.46 74.14 69.04 70.16

THI was calculated by using the following equation (0.8 × Ta) +[(RH/100) × (Ta – 14.4)] + 46.4 .

Fig. (2): Means of temperature humidity index (THI) during the experimental period .


35

The average ambient temperature during the experimental period ranged

from 38.3 o C to 45o C at 02:00 pm and 25.2 o C to 30.3 o C at 10:00 pm. While,

the respective average RH were 10% and 25%.

The average values of THI were between 79.43 to 85.46 at 02:00 pm and

69.04 to 74.14 at 10:00 pm during the experimental period .

The values of temperature humidity index THI from May to September in

New Valley recorded more than 74, indicating that animals under heat stress (kassab et al. 2017). Temperature humidity index value of 68 is considered the

upper limit of cattle comfort zone reported by (Johnson et al., 1989; Marai and

Habeeb, 2010). Temperature humidity index value of 74 to 78 is considered

hazardous and represents an alert condition for animals (Abd El-Ghany et al.,

2010). The present data indicated that animals were under heat stress during day and within the thermoneutral zone during night according to Davis et al. (2003).

4-2-Effects of antioxidant on thermoregulatory responses of experimental

cows .

Decreased animal performance under high ambient temperature is a cause

of concern in tropical and subtropical areas (Nardone et al., 2010; Katiyatiya et

al., 2015; Katiyatiya and Muchenje, 2017) as this triggers heat stress. Heat

stress reduces the efficiency of animal production leading to multibillion dollar losses to global animal agriculture (Bernabucci et al., 2010).

Heat stress, according to Hansen, (2009) is an environment that acts to

drive body temperature above set-point temperature. He also reported that body

temperatures of mammals usually range between 35–39 °C and they can be

maintained by the regulation of heat produced by the animal and heat loss to its

environment through the processes of convection, conduction and radiation. To

add on, each animal has a comfort zone beyond which it can experience stress


36

and a need will arise for thermal balance to be maintained (Nardone et al.,

2006).

The general homeostatic responses to thermal stress in mammals included

elevated respiration rate, panting, drooling of saliva, reduced heart rates, profuse

sweating, and reduced feed intake (Silanikove, 1992).

Table. (4): Effect of treated cows with antioxidant on thermoregulatory responses (mean±SEM)

Sign T3 T2 T1 C Items

NS 38.59 ± 0.04 38.73 ± 0.05 38.73 ± 0.35 38.79 ± 0.07 RT (ºC) * 34.67 ± 1.03 b 37.30 ± 1.26 ab 34.80 ± 1.01 b 37.67 ± 0.96 a RR

(breath/min) * 52.23 ± 1.25 b 54.10 ± 1.12 ab 53.13 ± .016 ab 54.90 ± 1.18 a PR

(beat/min.) NS 33.34 ± 0.24 33.47 ± 0.27 33.53 ± 0.26 33.66 ± 0.26 ST (ºC) * 32.45 ± 0.30 b 32.73 ± 0.28 ab 32.46 ± 0.31 b 32.92 ± 0.26 a HT(ºC)

NS 35.21±0.13 35.27±0.14 35.38 ± 0.19 35.48±0.14 ET (ºC) a, b Means of the same raw in each item with different superscripts are significantly different (P<0.05). RR, Respiration rate ; PR, Pulse rate; RT, Rectal temperature ; ST, Skin temperature; HT, Hair temperature ; ET , Ear temperature , C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

Previous studies have shown that high ambient temperatures amplify body

heat dissipation, leading to increased respiration rate and body temperature

(Marai et al., 2007; Sejian et al., 2010). Indeed, this most likely applies to any

of the dietary strategies that may be used to ameliorate heat stress, and so the

ability to predict when a heat stress event is likely to occur through web-based

tools or meteorological bureau data will greatly increase the ability to use dietary manipulations to ameliorate the effects of heat stress (Dunshea et al.,

2013). Hence, we accepted the hypothesis that antioxidant supplementation can

reverse some of the negative effects of heat stress on oxidative status and

improve some of the physiological responses to heat stress in cows.


37

Data in Table (4) and Figure (3 , 4) showed that using antioxidant agents

led to decrease rectal temperature , skin temperature and ear temperature, but

the differences were not significant. While, caused a significant decrease in

respiration rate, pulse rate and hair temperature in the treated animal.

These results demonstrate that using Vitamin E and Selenium or Zinc

sulphate or their combination alleviated the thermoregulatory responses to heat

stress in Aberdeen Angus Cows .

Fig. (3): Effect of treated cows with antioxidant on rectal temperature, respiration rate

and pulse rate of experimental cows

Recent research had shown that supplementation with vitamin E or

selenium, or their combination alleviated the physiological response to heat stress in dairy cows (Calamari et al., 2011), goats (Sivakumar et al., 2010)

and sheep (Alhidary et al., 2012 and Chauhan et al., 2016 a,b).

Supplementation of vitamin E and selenium, either individually or combined, at

supranutritional levels can partially mitigate against the effects of heat stress,

lowering respiration rate and rectal temperature and improving feed intake and oxidative balance in sheep (Sejian et al., 2014; Chauhan et al., 2016a).

Previously, there was no significant effect of dietary antioxidants on rectal or

skin temperature during heat stress despite the improvement in respiration rate.

(Chauhan et al., 2014).


38

In an elegant study, Sejian et al. (2014) included that supplemental

inorganic selenium and vitamin E, to Malpura ewes reduced the respiration

rate (−18%) and rectal temperature (−71%) . Respiration rates of cows in this

study were increased during heat stress in an effort to dissipate heat through the

respiratory tract. Interestingly, combined using of antioxidant reduced this

increase in respiration rate, indicating a reduction in the severity of heat stress.

Supplementation of vitamin E and selenium decrease the respiration rate, indicating a reduction in the severity of heat stress ( Chauhan et al ., 2014 ) .

Rectal temperature is another important indicator of thermal balance and

may be used to assess the adversity of thermal environment. A rise of 1°C or

less in rectal temperature is enough to reduce performance in most livestock

species, which makes it a sensitive indicator of physiological response to heat stress (Nardone et al., 2010 : Berman, 2011) as it is nearly constant under

normal conditions (Silanikove, 2000). Khalifa et al. (2016) indicated that

selenium supplementation either with vitamin E or in organic form did not affect

significantly rectal temperature and respiratory rates of Holstein dairy cows. The

decreased rectal temperature and respiratory rates in treatment groups indicated

that supplementation of vitamin E with selenium and vitamin C ameliorated the heat stress in goats,(Sivakumar et al., 2010). Similar decrease in rectal

temperature and respiratory rates by vitamin E and C supplementation was reported by Shenglins et al. (2003) in pigs and Kobeisy, (1997) in goats under

heat stressed conditions. Chirase et al. (1991) reported that steers fed control diet had higher

(P<0 . 10) mean rectal temperature than the steers fed Zn supplementation. In

addition Daghash and Mousa, (2002) showed that the significant decrease in

rectal temperature was recorded in buffalo calves after Zn supplementation.

The effects of heat stress on the heart rate has not been reported so far in

cows. However, it was expected to rise in response to increasing temperature in

the heat stress room in an effort to increase the blood flow towards the extremity


39

to dissipate heat from the body as was indicated by the increased skin

temperature. Interestingly, the heart rate was reduced by the antioxidant

supplementation, which may be explained by the beneficial effects of vitamin E

on the cardiac autonomic nervous system as has been reported in rats (Behrens et al., 1986; Fahim et al., 2013)

Fig. (4): Effect of treated cows with antioxidant on skin temperature, hair temperature

and ear temperature of experimental cows .

Skin temperature is another physiological parameter that can be used for

heat stress assessment as the heat exchange between the body and environment

is achieved through the skin. The adjustment of skin blood flow to regulate

transfer of heat from body core to the skin result in a shift in skin temperature in response to elevated temperatures (Habeeb et al., 1992).

Little data was available about effect of dietary antioxidant on ear temperature; Khalifa et al. (2000) found in goats, that exposure to heat stress

increased significantly (P<0.01) ear temperature by 3.0°C. They explained that

the increase in ear temperature under heat stress indicating the occurrence of

vasodilatation to facilitate in heat loss.


40

4-3- Effects of antioxidant on blood hematological parameters in

experimental cows.

4-3-1 - White blood cells and its fractions :

The leukocyte count is an important index for diagnosis of the health

status and problems, and may be adopted for assessing physiological changes

related to immune response (Bike, 2003). The effect of zinc sulfate

administration, vitamin E and selenium (Se) injection on white blood cells are

presented in Table (5) and Figure (5).

Results indicated that the average values of white blood cell (WBC) and

lymphocytes (LYM) of cows injected with vitamin E and selenium (T1) was

significant (P<0.05) higher than other treatments , while granulocytes (GRAN)

and monocytes (MID)did not exhibit any significant differences. These result

are agree with similar response in Ossimi lamb by (Soliman, 2015 b) who

reported that leucocytes profile showed a marked increase (P<0.01) in

lymphocytes percent for lambs received T1 and T2 vs. control. While the

differences in granulocytic and monocytes percentages were not significant. The

enhanced effect of vitamin E and selenium to increase lymphocytes agree with similar response in Ossimi sheep (Soliman et al., 2001), Awassi rams (Ammar

et al., 2009) and buffaloes (Qureshi et al., 2001).

The increase in blood lymphocyte populations may be a good indicator of an immunomodulatory response (Qureshi et al., 2001) . Mohri et al. (2005)

reported that Se and vitamin E supplementation affected on WBC counts in third

weeks of calves‘ life. In other study Kachuee, (2011) reported that the WBC

counts, neutrophils and lymphocytes counts were higher (P<0.05) in kids of

goats in selenomethionin group compared with the controls on birth day and 7

days of age. It has been explained that the higher leucocytes and lymphocyte cell

counts due to selenium and vitamin E administration could be related to the

protection of cell membrane and intracellular organelles by the effects of

selenium and vitamin E and thus increase their lifespan (Moeini and Jalilian,


41

2014) . Similar response of leukocytes counts to vitamin E plus selenium was also found in adult buffaloes (Qureshi et al., 2001), Friesian heifers

(Suwanpanya et al., 2007); and dairy calves (Mohri et al., 2005). In contrast,

supplementation of selenium alone or vitamin E plus Se had no effect on total count of leucocytes in buffalo calves (Shinde et al., 2009). While, Fazaeli and

Talebian, (2009) showed that the combination of vitamin E and Se were more

effective than selenium supplement to improve immune system . Table (5): Effect of treated cows with antioxidant on white blood cell (WBC) and its

Sign T3 T2 T1 C Parameters

* 8.88±1.89ab 8±1.15b 11.13±1.44a 8.18±1.35b WBC (103/μl)

* 2.88±0.33b 3.1±0.34 ab 4.01±0.43a 2.78±0.24 b LYM(103/μl)

NS 1.18±0.22 1.14±0.13 1.57±0.24 1.07±0.15 MID(103/μl)

NS 4.83±1.48 3.76±0.89 5.55±1.27 4.33±1.15 GRAN(103/μl)

a,b Means in the same row lacking a common superscript differ (P<0.05) WBC, White blood cell ; LYM, Lymphocytes ; GRAN, Granulocytes Cells ; MID, Monocytes C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

Fig. (5): Effect of treated cows with antioxidant on WBCs and its fractions .

The synergistic effect of vitamin E and Se to enhance lymphocyte function and its proliferative responses was detected with cattle (Pollock et al.,

1994). In in vitro study, exposure of peripheral blood lymphocytes to vitamin E

and Se treatment enhanced lymphocyte proliferation, indicating that this


42

response may optimize resistance to diseases (Ndiweni and Finch, 1995). The

increase in blood lymphocyte populations may be a good indicator of an immunomodulatory response (Qureshi et al., 2001). Injection of vitamin E

alone enhanced lymphocyte populations in calves (Reddy et al., 1986).

However, supplementation of Se alone at different levels in buffalo calves did

not alter the lymphocyte % (El-Ayouty et al., 1996). So, it appears that the

usefulness of Se to the immune function is increased by giving vitamin E

(Nockles, 1988); and the combination of vitamin E and Se were more effective

than Se alone to improve immune system (Fazaeli and Talebian, 2009). In sheep, Faixova et al. (2007) reported that supplementation of lambs with Se-

yeast had no positive effect on WBC counts.

In this study, dietary zinc had no significant differences WBC vs control . this result agree with Rupic et al. (1998) who reported that dietary Zn of organic

and inorganic origin had no effect on the WBC and platelet counts of fattening pigs . On another hand, Aliarabi et al. (2015) reported that White blood cells

and LYM differed significantly (P<0.05) between control and Zn-supplemented

groups they reported that Zn supplementation increased these parameters compared to control Also, Sobhanirad et al. (2014) found that the number of

WBC of lamds treated with 50 mg/kg zinc propionate and 50 mg/kg zinc

sulphate were significantly high (P<0.05) compared to the other groups. In the

other study, Southern and Baker, (1983) found that White blood cell and LYM

differed significantly(P<0.05) between control and Zn-supplemented groups Zn

supplementation increased these parameters compared to control.

4 -2- Red blood cells , Hb and HCT

Data in Table (6) and Figure (6,7) showed that no significances

differences were found among treatment in red blood cell (RBC) counts.

Treated cows ( T1) had higher (P<0.05) concentration of hemoglobin

concentration (Hb), hematocrit (HCT), red blood cell distribution width (RDW)

and mean corpuscular volume (MCV) than those of T3 , control and T1 . The


43

results revealed that mean corpuscular hemoglobin (MCH) increased (P<0.05)

for experimental cow compared to a control. While, the differences in mean

corpuscular hemoglobin concentration (MCHC) were not significantly. Value of RBCs tended to decrease in the T2 group and this accordance with El-Masry et

al. (1998). Low estimates for RBCs were reported by Ukanwoko et al. (2013),

whereas high value of RBCs were showed by Waziri et al. (2010), Kwari et al.

(2011) and Olorunnisomo et al. (2012). Table (6): Effect of treated cows with antioxidant on red blood cell (RBC), Hb and HCT

(mean±SEM)

Sign T3 T2 T1 C Parameters

NS 6.84±0.19 6.44±0.26 6.94±0.38 6.59±0.24 RBC (106/μl) ** 11.63±0.39 ab 10.65±0.37 b 12.84±0.81 a 10.73±0.36 b Hb (g/dl) ** 31.67±1.08 b 28.53±1.06 b 34.99±1.95 a 30.38±1.04 b HCT %

** 48.21±0.97 a 42.76±1.63 b 50.37±0.29 a 43.09±1.68 b MCV (fl) ** 17.78±0.64 ab 15.92±0.45 c 18.52±0.43 a 16.58±0.4 bc MCH (pg) NS 36.79±0.66 35.44±0.57 37.33±0.27 36.73±0.8 MCHC (g/dl) ** 31.98±0.98 a 29.25±0.61 b 33.63±0.49 a 29.86±0.61 b RDW

a,b,cMeans in the same row lacking a common superscript differ (P<0.05) RBC, red blood cell ; HB, hemoglobin concentration ; MCH, mean corpuscular hemoglobin ; MCHC, mean corpuscular hemoglobin concentration ; HCT,hematocrit ; MCV, mean corpuscular volume; RDW, red blood cell distribution width. C: Control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

Fig. (6): Effect of treated cows with antioxidant agents on RBC and HB .

Dietary zinc had no effect on Hb. This result were agree with Kwari et

al. (2011) and Ukanwoko et al. (2013). lower estimate of Hb was reported by


44

Olorunnisomo et al. (2012), while the higher Hb was reported by Waziri et al.

(2010) .

Cows of T1 group had higher (P<0.05) concentration of Hb than other

treatments ,These results supported the positive effect of vitamin E and Se

supplementation on blood hematology as reported in earlier studies in sheep

(Soliman et al., 2001; Makkawi et al., 2012), cattle calves (Mohri et al., 2005)

and buffaloes (Qureshi et al., 2001) .

Fig. (7): Effect of treated cows with antioxidant on HCT, MCV, MCH , MCHC and

RDW .

In the present study, some hematological parameters were positively

changed up as a result of vitamin E plus Se treatment, which may point out the

active metabolism and biological oxidation effects on the cellular base that

might lead to availability of metabolites required for tissue growth. The

significant increase in blood Hb due to injection of vitamin E plus Se agree with the results reported on growing Ossimi lambs (Soliman et al., 2001) and dairy

calves (Mohri et al., 2005). Likewise, these findings are in accordance with the

study of Qureshi et al. (2001) who found a significant higher Hb, RBC counts

and HCT values, while MCV, MCH and MCHC remained unchanged for adult

buffaloes supplemented with vitamin E and Se during late gestation. The lambs


45

born from vitamin E plus Se-injected ewes, had a significant increase in blood

Hb and MCHC and showed tendency to increase MCH values. In contrary to the

above results, supplementation of Se and vitamin E in the diet of buffalo calves

had no significant effect on hematological parameters (Hb, HCT, RBCs and WBCs counts) as reported by Shinde et al. (2009).

Sobhanirad et al. (2014) showed that the HCT and MCV in 50 mg/kg

zinc propionate (P<0.05) than other groups. Number of RBC, Hb concentration,

MCH and MCHC were higher (P<0.05) in 100 mg/kg zinc propionate compared with other groups. Bremner et al. (1976) reported that higher level of

Zn (420 mg/kg) induced depression in haematological parameters such as blood

Hb concentration and HCT when compared with diets providing 48 mg/kg Zn in

sheep.

4-4-Effects of antioxidant on blood biochemical parameters in experimental cows: 4-4-1- Total protein The effect of zinc sulfate administration, and vitamin E + selenium (Se)

injection on blood metabolites are presented in Tables (7, 8, 9) and Figures (8,

9, 10, 11) .

Results indicated that the average values of total protein in blood serum

for cows did not exhibit any significant differences among treatments and

control. Results in this study in cows injected with vitamin E plus selenium agree with that reported by Khalifa et al. (2016) who reported that Se

supplementation either with vitamin E or in organic form had no significant effect on plasma total protein of Holstein cows. Also, Shinde et al. (2009)

found no effect on serum total protein with supplementation of selenium and

vitamin E in the diet of buffalo calves. While El-Shahat and Abd El-monem,

(2011) found that ewes received selenium had a significant low serum total

protein . On another hand Soliman et al. (2012) found that injected ewes with


46

vitamin E plus selenium at late gestation and during suckling period had a

significant increases in plasma total protein. Table. (7): Effect of treated cows with antioxidant agents on total protein and its

fractions (mean±SEM) Sign T3 T2 T1 C Items

NS 7.56 ± 0.1 7.44 ± 0.08 7.66 ± 0.13 7.57± 0.1 Total protein (g/l)

* 3.13 ± 0.06ab 3.25 ± 0.07ab 3.31 ± 0.05a 3.07 ± 0.11c Albumin (g/dl)

NS 4.43 ± 0.14 4.2 ± 0.14 4.35 ± 0.14 4.5 ± 0.2 Globulin (g/dl) a,b,cMeans in the same row lacking a common superscript differ (P<0.05) C: Control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc

sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

Fig (8) Effect of treated cows with antioxidant on on total protein, albumin and globulin

Hassan et al. (2016) stated that zinc sulfate supplementation to growing

buffalos calves did not significantly effect in serum total protein concentration . Similarly, Farghaly et al. (2017) reported that serum concentrations of total

protein were not affected by zinc supplementation in growing lambs . While,

Daghash and Mousa, (2002) reported a significant increase of total protein in buffalo calves after Zn supplementation. On another hand El-Hendy et al.

(2001) reported that in two test groups fed with insufficient zinc diet, total

protein was significantly lower than controls .


47

4-4-2. Albumin Data in Table (7) and Figure (8) showed that the serum albumin

concentrations of cows injected with vitamin E plus selenium and zinc sulfate

supplementation were significantly (P<0.05) higher than control. These results are agreement with those reported by Daghash et al. (1993) who observed that

serum albumin tended to be higher in group injected with Vitamin E than control. Also, Metry et al. (1998) found that treated buffalo calves with Vitamin

E had more obvious effect (P<0.01) on albumin value. In addition, Soliman,

(2015a) showed a significant increase in serum albumin concentrations for

lambs received vitamin E and vitamin A+E treatments compared to those received vitamin A alone . On another hand, Khalifa et al. (2016) did not find

significant effect of Se supplementation on plasma albumin levels in Holstein

cows. Also, Soliman et al. (2012) found that ewes injection with vitamin E at

late gestation and during suckling period had no significant change in plasma

albumin in compared with control ewes.

Results in the present work showed that level of serum albumin

concentration of cows supplemented Zinc sulphate are agreement with those

reported by Shakweer et al. (2010) who reported that albumin concentration

was increased blood plasma with zinc sulfate addition in of Friesian calf . Similarly, Fagari-Nobijari et al. (2012) observed that albumin was higher (P<

0.05) in young Holstein bulls fed ZnSO4 than control . While, Mousa and EL-

Sheikh, (2004) indicated that zinc sulfate addition decreased albumin in blood

serum of buffalo calves. In other study, Hassan et al. (2016) revealed that there

were no significant effects in concentrations of serum albumin due to zinc supplements for calves. Also, Farghaly et al. (2017) reported that serum

concentration of albumin was no affect by zinc supplementation in growing

lambs .

4-4-3. Globulin


48

Data in Table (7) and Figure (8) showed that the serum globulin

concentration of cows injected with Vitamin E plus selenium and zinc sulfate

supplementation did not exhibit any significant differences among treatments

and control . Similar globulin response was reported by Shinde et al. (2009) who

reported that no effect of supplementation of selenium and vitamin E on serum globulin for buffalo calves. Also, Daghash et al. (1993) observed that serum

globulin concentrations did not differ as a result of vitamin E injection . In

cattle, feeding vitamin E (4400 IU/d) increased serum albumin fraction but did not affect different fractions of globulin (Rahmani et al., 2014). Similar

responses were detected in buffaloes (Helal et al., 2009). Also, Kassab and

Mohammed, (2014) reported that serum globulin concentrations did not

changed due to vitamin E plus Se treatment of Sohagi ewes . While, Soliman,

(2015b) indicated that lambs gave high dose of vitamin E and selenium led to

higher (P<0.05) serum globulin than those gave low dose of it. Also, Shetaewi et al. (1992) reported that supplemented vitamin E at dose 100 and 200 mg

increased serum globulin compared to control .

Results in the present work showed that level of serum globulin

concentration of cows supplemented zinc sulphate are agreement with those reported by Hassan et al. (2011) who reported that inorganic or organic zinc did

not cause any effect on the serum globulin in Barki ewes. Also, Farghaly et al.

(2017) revealed that there were no significant differences (P>0.05) in serum globulin of lambs due to Zn-supplemented diet. On another hand , Bednarek et

al. (1991) found a significant increase in gamma globulin in calves fid diet

supplemented with zinc .Also, Bires et al. (1992) found in lactating cows that

Zinc treated group had higher blood immunoglobulin , than those in untreated

group . In buffalo calves, Daghash and Mousa, (2002) reported a significant

increase of total globulin after Zn supplementation .


49

4-4-4. Glucose

Results presented in Table (8) and Figure (9) showed that the serum

glucose concentration was not significantly (P<0.05) differences among cows

with T1, T2 , T3 and control . the presented results agree with similar response in lambs by Soliman et al. (2012) who reported that no significant changes were

observed in plasma glucose either for ewes treated with vitamin E and selenium or their lambs . Also, Meydani et al. (1994) reported that vitamin E

supplementation had no effect on plasma glucose. Also, Soliman, (2015b)

found that injection of various doses of vitamin E and selenium had no

significant effect on serum glucose concentrations in lambs. Similar result of

blood glucose was observed supplemented with vitamin E plus selenium in sheep (Soliman et al., 2001) and in goats supplemented with selenium

(Kamdev et al., 2015).

In sheep, Alhidary et al. (2015) observed increase in the serum

concentrations of glucose fed diets supplemented with selenium and vitamin E .

On another hand , A decrease in plasma glucose concentration from 3.09 to 2.76

mM as result of supplementing the diets of calves with Se (0.3 mg Sel-Plex per

kg DM) has been reported (Ebrahimi et al., 2009) . Singh et al. (2002)

observed low blood glucose concentration in buffalo calves fed wheat straw

containing high selenium. The present results are agree with those reported by Farghaly et al.

(2017) who found that no significant differences in serum glucose of lambs

supplemented with zinc . As the same Aliarabi et al. (2015) reported that

plasma glucose concentration did not differ significantly between control and Zn supplemented groups of lambs . However, Hassan et al. (2011) reported that

organic Zn caused a significant(P < 0.05) decline in glucose compared to the

inorganic Zn . Table (8) Effect of treated cows with antioxidant on Glucose, Cholesterol, AST and

ALT (mean±SEM)


50

Sign T3 T2 T1 C Items

NS 57.83 ± 1.03 56.00 ± 1.71 59.00 ± 1.55 56.83 ± 1.6 Glucose (mg/dl)

** 155.2 ± 12.83b 151.5 ± 7.68b 190.1 ± 8.91a 153.9 ± 8.72b Total Cholesterol (mg/dl)

* 47.58 ± 3.14ab 40.92 ± 2.48c 50.33 ± 2.70a 43.92 ± 3.05bc AST (U/l)

NS 18.33 ± 1.94 17.00 ± 1.88 19.53 ± 1.58 18.17 ± 2.02 ALT (U/l) a,b,c Means of the same row in each item with different superscripts are significantly different (P<0.05) C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate AST, aspartates aminotransferase ; ALT, alanine aminotransferase 4-4-5. Cholesterol

Data in Table (8) and Figure (9) showed that cholesterol concentration of

cows treated with vitamin E and selenium (T1) was significantly (P<0.05)

highar than T3, T2 and control groups , with no significant defferances were

abserved among T3 , T2 and control groups. Results of cholesterol

concentration were in the normal range (50 – 250 mg/dl) as reported by

Mansourian, (2010) The results are agreement with Nayyar et al. (2003) who reported that

supplementation of vitamin E and vitamin E + selenium to anoestrous buffalo

heifers increase significantly (P<0.01) cholesterol concentration compared to

unsuppleemnted groups. Similarly , Avei et al. (2000) found that supplement

ewes with vitamin E 250 mg\kg increased plasma cholesterol level than those

supplement with 1500IU/kg, while the minimum value recorded in the control group . Rahimani et al. (2015) Found that treatment of cows at early lactation

with vitamin E, beginning 5 weeks postpartum was more effected on cholesterol

concentration. In contrast, Soliman, (2015b) found that serum cholesterol

concentration decreased (P<0.05) for lambs received two doses of vitamin E vs.

control. In contrast, Ziaei, (2015) reported that supplementation of diets with Se or vitamin E had no significant effect on serum cholesterol . Also, Metry et

al. (1998) indicated that serum cholesterol levels not affected by


51

supplementation with vitamen E alone or combined with selenium for Egyptian

buffalo calves .

Fig. (9): Effect of treated cows with antioxidant on glucose, cholesterol, AST and ALT

Acocording to zinc sulphate , results showed that cows fed diet containing

zinc did not differ in cholesterol concentration among T2 , T3 and the control group . This results are agreement with El-Amin et al. (2013) who showed that

dietary Zinc supplementation did not affected significantly (P>0.05) the levels of serum cholesterol. On the other hand Farghaly et al. (2017) reported that

serum cholesterol concentration of lambs fed diets supplemented with Zn-SO4

or Zn-Meth were higher (P>0.05) than that lambs fed the control diet .

4-4-6 Liver function (ALT and AST enzymes )

4-4-6-1 Aspartate Amino Transferase (AST).

Results in Table (8) and Figure (9) indicted that mean value of AST was

no significant difference in T2 group in comparison with the control one .

While, the activity of AST increased significantly (p<0.05) in T1 group compare

with the control group. However , no significant difference was found between

T3 and T1 groups. Regarding selenium supplementation, Singh et al. (2002) observed that

buffalo calves fed high Se (8.54 ppm) had higher activity of plasma AST. Also ,


52

Shashidhar and Prasad, (1993) found an increase of serum AST activity when

adult goats supplemented with 0.15 – 0.30 mg Se per kg body weight. Contrary to present findings, Cronje et al. (2006) reported that concentrations of AST in

sheep injected daily with 1.6 mg Se were less than in untreated sheep (48 vs. 96

and 40 vs. 87 units/L, respectively;) . Also Soliman. (2015) found that

concentrations of AST in sheep injected with different doses of vitamin E and selenium were less than control . On the other hand , Shinde et al. (2009)

reported that the activity of AST did not differ among buffalo calves

supplemented with selenium, and vitamin E + selenium and control group Also, Samanta and Dass, (2007) and Mudgal et al. (2008) did not find any effect of

supplemental vitamin E and selenium on the activity of AST in buffalo calves. Pond et al. (1995) did not observe any difference in serum AST in goats fed the

basal diet alone or supplemented with either inorganic or organic selenium.

Similar response of serum AST enzyme activity due to selenium supplementation was observed in goats (Kamdev et al., 2015). Moreover,

Kassab and Mohammed, (2014) reported that non-significant effect in AST

activity of Sohagi ewes injected with vitamin E and selenium.

According to zinc sulfate, results showed that the activity of AST did not differ between T2 and control one. This results agreed with Hassan et al. (2016)

they reported that no significant differences were observed of AST with Zn supplements for calves. Also, Hassan et al. (2011) reported that inorganic or

organic zinc did not cause any effect on the serum AST in ewes . Also, Mandal et al. (2008) reported that no differences in AST activity were observed due to

Zn level in crossbred calves. Contrary to these findings, Daghash and Mousa,

(1999) observed higher AST activity in buffalo calves supplemented with 50 or

100 ppm Zn.

4-4-6-2. Alanine Amino transferase (ALT) .


53

Results in Table (8) and Figure (9) indicted that mean values of alanine

amino transaminase (AST) did not differ among groups. This result is in agreement with those obtained by Shinde et al. (2009)

who reported that no significant variation the activities of ALT of buffalo calves

supplemented with vitamin E + selenium and control group. Also, Samanta and

Dass, (2007) and Mudgal et al. (2008), did not find any effect of supplemental

vitamin E and selenium on the activity of ALT in crossbred and buffalo calves.

Also, Kassab and Mohammed, (2014) reported no significant effect in the

activities of ALT of Sohagi ewes injected with vitamin E and Selenium. On the

other hand, Surai, (2006) indicated that increases in the activities of ALT in

sheep and cattle with a diet deficient in selenium and vitamin E.

The present results indicated that cows fed diet supplemented with zinc

sulfate did not differ the activities of ALT. This results are in agreement with those reported by Hassan et al. (2011) who reported that inorganic or organic

zinc did not cause any effect on the serum (ALT) in ewes . Also, Mandal et al.

(2008) reported that no differences in ALT activities were observed due to Zn level in crossbred calves . In contrast, Shinde et al. (2013) found that ALT

activities significantly (p<0.05) declined in Cu- and Zn-met group than Cu- and

Zn sulphate indicating an effective role of chelated minerals in improving liver

function .

The concentration of AST and ALT recorded in the present study are in

agreement with the normal ranges of AST and ALT activities (U/l) recorded previously in cattle by Stojevic et al. (2005). They reported that the normal

ranges of AST and ALT activities are 19.2 to 84.90 and 4.2 to 29.7, respectively.

Thus, the present results indicated that using antioxidant did not affect the

physiological functions of the important organs, practically the liver function.

4-4-7. Kidney function (Urea-N and Creatinine):

Urea-N and creatinine concentrations in blood serum of cows as affected

of the experimental treatment are presented in Table (9) and Figure (10) ,


54

Results indicated that cows treated with zinc sulfate administration, vitamin E

and selenium injection or both did not affect serum urea-N and creatinine

concentrations compared with control one . Table (9): Effect of treated cows with antioxidant on urea-N, creatinine , and Total antioxidant capacity (TAC) (mean±SEM)

Sign T3 T2 T1 C Items NS 22.04 ± 1.62 23.66 ± 1.93 24.15 ± 1.52 24.88 ± 1.74 Urea-N (mg/dl) NS 1.83 ± 0.09 1.91 ± 0.08 1.78 ± 0.12 1.84 ± 0.07 Creatinine (mg/dl) NS 0.65 ± 0.07 0.61 ± 0.12 0.62 ± 0.04 0.57 ± 0.07 TAC (mmol/L)

C: Control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate TAC , total antioxidant capacity

Fig. (10): Effect of Effect of treated cows with antioxidant on Urea-N and Creatinine

This results are agree with those reported by Kassab and Mohammed,

(2014) who found that no significant effect in serum urea and creatinine of

Sohagi ewes injected with vitamin E and selenium. Also, Shinde et al. (2009)

reported that non-significant variation in serum urea-N and creatinine

concentrations of buffalo calves supplemented with selenium, and vitamin E + selenium. Also, Mudgal et al. (2008) reported that supplementation of selenium

to buffalo calves had no effect on serum urea-N and creatinine levels.

Since serum urea-N and creatinine are indicators of the normal

physiological status and nitrogen metabolism of the animals, Hence it can be


55

concluded that supplementation of selenium, vitamin E or both had no effect on the nitrogen metabolism of buffalo calves (Shinde et al., 2009) .

Result in the present study in cows supplemented with zinc sulfate agree with that reported by Hassan et al. (2011) who reported that inorganic zinc

(ZnSO4·7H2O) did not cause any effect on serum creatinine and urea-N

concentrations. Also, Sobhanirad et al. (2014) reported that supplementation of

different source and level of zinc to Baluchi lambs had no effect on serum urea-

N levels. On the other hand , Mousa and EL-Sheikh, (2004) indicated that zinc

sulfate addition decreased urea-N concentration in blood serum of buffalo calves. Also, Zeedan et al. (2014) reported a significant decrease in plasma

urea-N and creatinine concentrations in dairy Damascus goats supplemented

with Biogen–Zinc . While, Aiello, (1998) reported that increase in serum

creatinine concentration was associated with selenium and vitamin E

supplementation.

4-4-8 Total antioxidant capacity (TAC)

Total antioxidant capacity (TAC) measures the antioxidant capacity of

antioxidants in a biological sample (Kusano and Ferrari, 2008).

The effect of zinc sulfate administration, vitamin E and selenium (Se)

injection on total antioxidant capacity (TAC) are presented in Table (9) and

Figure (11) . There were no significant differences in TAC among groups.

However, the highest (TAC) was recorded for cows treated with zinc sulphate

and vitamin E + sselenium (T3 ) followed by those groups treated with vitamin

E + selenium (T1) , at the same time zinc sulphate (T2 ) led to lesser value

compared to control.


56

Fig. (11): Effect of treated cows with antioxidant on Total antioxidant capacity (TAC)

The present results are in agreement with those reported by Rashidi et al.

(2011) who indicate that supplementation with Zn, vitamin E or both did not

affect serum TAC in any of the studied groups compered to control one . On the

other hand, selenium and vitamin E increases plasma concentrations of total antioxidant capacity and Se in dairy cattle (Calamari et al., 2011) , sheep

(Alhidary et al., 2015) and goats (Katamoto et al., 1998) . Also, Khatti et al.

(2017) found that treated cows with vitamin E and selenium increased (P < 0.05)

TAC levels. Moreover, in a study, adding vitamin E (80 IU/ kid/d) to Boer goats increase serum TAC (Hong et al., 2010) . In an elegant study on Holstein cows ,

supplementation of vitamin E did not affect TAC (Rahmani et al., 2014) .

Similar to our results Farghaly et al. (2017) reported that serum total

antioxidant capacity (TAC) of lambs fed diets supplemented with Zn-SO4 were higher (P>0.05) than that of lambs fed control diet. Also , Aliarabi et al. (2015)

reported that total antioxidant capacity (TAC) concentration did not differ

significantly with Zn-supplemented treatment (P>0.05) , However the average

value of serum TAC of lambs fed diets supplemented with Zn-SO4 or Zn-

proteinate were higher (P>0.05) than that of lambs fed control diet . On another hand Hassan et al. (2011) fonund that inorganic Zn (ZnSO4·7H2O) caused a


57

significant decline (P < 0.05) in the antioxidants activity compared to organic Zn

(Zn.Met) .

Results showed that antioxidant concinteration increased by

supplementation of vitamin E with selenium and zinc indicating improvement in

the antioxidant activity and decrease oxidative stress to cows.

4-5- Effects of using antioxidant on reproductive performances of

experimental cows.

Data in Table (10) illustrated that treated cows with antioxidant showed

better reproductive performance in all reproductive parameters considered in the study than control group. Also Khan et al. (2015) reported that mineral and

vitamin E supplementation improved the reproductive performances of buffalo

during periparturient period. Campbell and Miller, (1998) also reported

improvement of reproductive performances for dairy cows supplemented with

vitamin E and Zn.

4-5-1- Response to synchronization

Estrus synchronization is a beneficial tool for cattle producers due to its

ability to shorten the breeding season and create a more concise calving season (Larson et al., 2010). In the present study, there was no any significant effect

of treatments on estrus synchronization response, Table (10). These results are

in agreed with those reported by Brasche, (2015) who reported that heifers

exhibited no difference in their response to synchronization due to trace mineral

containing copper (Cu), manganese (Mn), selenium (Se) and zinc (Zn) . Moreover, Makkawi et al. (2014) reported that there was no difference in their

response to synchronization due to injection vitamin E and selenium in ewe .

On the other hand, Koyuncu and Yerlikaya, (2007) found that selenium and

selenium plus vitamin E treatments had a significant beneficial effect on oestrus

response in ewes if compared to control. Furthermore, Kundu et al. (2014)


58

indicated that the supplementation of different levels of inorganic zinc oxide

significantly increased the incidence of oestrus of Teressa goat.

Table (10): Effect of treated cows with antioxidant on reproductive performance .

Sign T3 T2 T1 C Items --- 4 4 4 4 Number of animals NS 100 100 100 100 Percentage of estrus % ** 75 (3/4)a 75 (3/4)a 75 (3/4)a 25 (1/4)b First service conception rate % ** 100(1/1)a 100(1/1)a 100(1/1)a 66.7(2/3)b Second service conception rate% ** 100(4/4) a 100(4/4) a 100(4/4) a 75 (3/4) b Conception rate % ** 1.25(5/4)b 1.25(5/4)b 1.25(5/4)b 1.75 (7/4)a Number of services per

conception Values in the same row with different superscripts (a, b ) are different (P<0.05) C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

4-5-2- First service conception rate

Data in Table (10) and Figure (12) showed that cows injected with

vitamin E and selenium (T1) , supplemented with zinc sulphate (T2) or that

supplemented zinc sulphate in combination with vitamin E and selenium (T3)

improved first service conception rates (75%) when compared to control group

(25%) .

Fig. (12): Effect of treated cows with antioxidant on first service conception rate

Selenium supplementation in selenium deficient dairy cows decrease the

number of service per conception and led to increase pregnancy rate at first service (Kommisrud et al., 2005). Furthermore, injection of vitamin E and

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4478481/#b23-ajas-28-8-1133


59

selenium to cows 3 weeks prepartum shortened calving to conception in carvel and increased pregnancy rate at first service from 25.3 to 41.2% (Arechiga et al.,

1994). In addition, Ballantine et al. (2002) reported a decrease in days open in

cows pregnant at first service conception risk in cows supplemented with Zn, Mn,

Cu, and Co complexes.

4-5-3- Conception rate

Data in Table (10) and Figure (13) showed that cows injected with

vitamin E and selenium (T1) , supplemented with zinc sulphate (T2) or that

supplemented zinc sulphate in combination with vitamin E and selenium (T3)

improved conception rates (100%) compared to control group (75%) .

Fig.(13): Effect of treated cows with antioxidant on conception rate

In the present study, injection of vitamin E and selenium and

supplementation of zinc sulphate in Aberdeen Angus cows improved the ovarian

activity and consequently the reproductive traits as monitored by conception rate

in comparison with the control group. The present result are concomitant with those reported by Khatti et al. (2017) showed an increase in the pregnancy rate

of crossbred cows supplemented with vitamin E and selenium. Also, Mavi et al.

(2006) stated that supplementation of vitamin E and selenium to Murrah

buffaloes during prepartum period has caused a significant increase in


60

conception rate . Moreover, improvement of fertilization rate could observed with vitamin E and selenium supplementation in cattle (Segerson et al., 1981)

and in sheep (Segerson et al., 1977).

Macpherson et al. (1987) reported that a single subcutaneous injection of

barium selenite improved conception rate significantly of Friesian heifers.

Furthermore, Nayyar et al. (2002) reported a significant increase in conception

rate in buffalo heifers supplemented with 14 mg of selenium per week.

Hemingway, (2003) reported an increase in conception rate by providing a

sustainedrelease multi-trace element rumen bolus in beef cattle. He attributed

this increase in conception rate due to the effect of selenium on establishment of

ova and sperm transport.

Earlier reports stated that reduction in pregnancy rates can occur as a subclinical effect of marginal trace mineral status (Hostetler et al., 2003).

Deficiencies in Cu, Se, Mn, and Zn have been linked to abnormal estrus cycles, impaired ovulation and decreased conception rates (Hostetler et al., 2003;

Underwood, 1981). In beef heifers that were provided a free choice

supplementation of Cu, Mn, and Zn had illustrated a greater conception rate to timed embryo transfer when compared to control heifers (Sales et al., 2011) and

higher conception rates to fixed timed artificial insemination (Stanton et al.,

2000; DeFrain et al., 2009; Mundell et al., 2012).

Kundu et al. (2014) indicated that goat that received 100 ppm ZnO

showed significantly (P<0.05) higher pregnancy rate (100%) followed by 50

ppm group (66.66%) and the control group (50%). On the other hand, Abdel-

Monem and El-Shahat, (2011 ) reported that inclusion of 50, 100, and 150 ppm ZnO increased pregnancy rates compared to controls. Ahola et al. (2004)

also found that cows received trace minerals (Zn, Cu and Mn) had higher

pregnancy rates than non-supplemented cows.


61

Another trial also reported a tendency for increased percentage of cows

pregnant at 150 DIM in cows fed a mixture of sulfates and Zn, Mn, Cu, and Co

complexes compared to similar levels of those minerals provided as sulfates (DeFrain et al., 2009) .

Zinc has a critical role in the repair of damaged uterine tissue and

maintenance of uterine lining following parturition, speeding return to the normal reproductive function and oestrus (Green et al., 1998). Improvement of

reproductive performances in dairy cows supplemented with Zn had reported by Manspeaker et al. (1987). This may be due to increase plasma β-carotene level

that correlates directly to the improved conception rates and embryonic

development (Hayat et al., 2010).

4-5-3. Number of services per conception

Data in Table (10) and Figure (14) showed that cows injected vitamin E

and selenium (T1) , supplemented with zinc sulphate (T2) or that supplemented

zinc sulphate in combination with vitamin E and selenium (T3) had decreased

the number of services per conception (1.25) compared to control group (1.75) .

Fig. (14): Effect of treated cows with antioxidant on number of services per conception.


62

Supplementation with selenium and vitamin E decreased the number of services per conception of dairy cattle as reported by Sattar et al. (2007) and

Bayril et al. (2015).

Dietary and /or injectable form of vitamin E supplementation to dairy cows decreased number services per conception (Jukola et al., 1996; Kim et al.,

1997). Moreover, injection of vitamin E and selenium to cows 3 weeks

prepartum reduced the number of services per conception from 2.8 to 2.3 (Arechiga et al., 1994). Previous reports by Archeiga et al. (1998) concluded

that injection of 500 mg of vitamin E and 50 mg of Se to 30 day postpartum

cows reduced days open (98.1 vs. 84.6 d, p<0.05) and services per conception

(2.0 vs. 1.7). On the other hand, Harrison et al. (1984) and Moenini et al.

(2009) reported that Se and Vit E supplementation had no effect on the number

of service per conception.

Feeding organic trace elements increased conception rates in cattle,

reduced days open (13.5 less days open) and number of services per conception (0.27 less services per conception) (Rabiee et al., 2010) and increased ovarian

activity in dairy cows (Boland, 2003). Supplementations of Cu, Zn and Mn as

well as Se and vitamin E had been shown to reduce services per conception in cattle (Ishak et al., 1983). Also, Nayeri et al. (2014) reported that feeding

complexed zinc decreased the number of services per conception in Holstein

cows regardless of level.

4-5-5 Size of embryo, amniotic vesicle and corpus luteum

Data in Table (11) and Figure (15) showed that size of embryo,

embryonic vesicle and corpus luteum were significantly (p<0.05) higher in T3

than other treatments.

During early fetal growth, Se functions as a reductase to help protection

against mortality from oxidative damage (Hostetler et al., 2003). Earlier reports

concluded that injecting beef cows with vitamin E and selenium produced more fertilized ova than control animals (Kappel et al., 1984).




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63

Table (11): Effect of Effect of treated cows with antioxidant on size of embryo (mm), Amniotic vesicle (mm) and corpus luteum. (mean±SEM)

Sign T3 T2 T1 C Items ** 17.37±0.30a 13.63±0.38b 14.53 ± 0.2b 12.8c Size of embryo (mm) ** 28.07±0.38a 21.17±1.30b 21.87±1.07b 19.6b Amniotic vesicle (mm) * 23.07±0.78a 21.07±0.78ab 22.03±1.07a 18.6b corpus luteum (mm)

Values in the same row with different superscripts (a, b, c) are different (P<0.05) C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

Some antioxidant systems actively work in the corpus luteum during

pregnancy (Sugino et al., 2000). It is known that vitamin E and selenium as

antioxidant could play a role in the antioxidant system in the corpus luteum

(Kamada and Hodate, 1998 ). The mechanism of action of Se as it degrades H2O2 or peroxides in the corpus luteum as a component of GSHPx (Flohe et

al., 1973) or phospholipid hydroperoxide glutathione peroxidise (Kamada and

Ikumo, 1997) which support its hormone production activity. Furthermore,

Kamada and Hodate , (1998) have reported the positive effects of Se on

plasma progesterone concentration production from luteal cells.

Fig. (15): Effect of treated cows with antioxidant on size of embryo (mm) and Amniotic vesicle (mm)


64

As the participation of zinc is centrally involved in most events relating to

cell division and nucleic acid synthesis repair including many other enzymatic

pathways, its presence is most important during periods of rapid cellular

development i.e. during organogenesis and fetal growth. Indeed, the time of

conception, and the following pregnancy, represents the most vital period for

assuring an optimum zinc status. In fact, it has been already established that

inadequate parental zinc status can result in a variety of deleterious effects on

the offspring. The most obvious connection between clinical and/or sub-clinical

zinc deficiency and fetal teratology seems to relate directly to its central role in

all processes concerning cell differentiation and replication. ( Tuormaa , 1995)

4-5-6. Ovarian follicular development and hormonal profile during estrous cycle

4-5-6-1 Follicular development

Antioxidants play a role in gonadotrophic hormone receptors protection form oxidation with subsequent elevation of oestrogen secretion (Sergerson et

al., 1980) which is important for follicular development. The

requirement/involvement of antioxidants is evident as a protective effect of

retinol on oocytes during heat stress (Lawrence et al., 2004).

The present results from the laparoscopic examination of ovarian activity

of Aberdeen Angus cows are illustrated in Table (12) and Figure (16) showed a

significant difference was found among different groups regarding the

population of small, medium and large follicles. The lowest population of

follicles found in this category was for control. Follicles population consisted

mainly of small follicles, followed by medium follicles, which were similar in

all studied groups. Thus all groups studied had the same potential of ovarian

activity in the reproductive stage.


65

Table (12): Effect of treated cows with antioxidant on number of different size of ovarian follicles during estrus period (mean ± SEM)

sign

Means of days

Treatments (T) Days Number of follicles T3 T2 T1 C

<0.0

001

2.33±0.19d 2.67±0.33 2.00±0.58 2.67±0.33 2.00 0 Small follicles (<5mm)

3.17±0.17c 3.67±0.33 3.00 3.33±0.33 2.67±0.33 3 2.50±0.15d 2.67±0.33 2.33±0.33 2.67±0.33 2.33±0.33 6 3.92±0.23b 4.67±0.33 3.67±0.33 4.00±0.58 3.33±0.33 9 3.17±0.24c 3.67±0.33 2.67±0.33 3.33±0.67 3.00±0.58 12 4.17±0.32b 5.33±0.33 3.67±0.88 4.33±0.33 3.33±0.33 15 4.00±0.21b 4.67±0.33 3.67±0.33 4.33±0.33 3.33±0.33 18 5.17±0.17a 5.67±0.33 5.00 5.33±0.33 4.67±0.33 0 4.13±0.24A 3.25±0.23B 3.75±0.22A 3.08±0.19A Means (T)

<0.0001 sign

<0.0

001

2.08±0.15c 2.33±0.33 1.67±0.33 2.00 2.33±0.33 0 Medium follicles (5-8 mm)

3.25±0.25ab 4.33±0.33 2.67±0.33 3.33±0.33 2.67±0.33 3 2.67±0.31bc 3.33±0.88 2.67±0.67 2.67±0.33 20.00±0.58 6 3.50±0.26a 4.00±0.58 3.33±0.67 3.67±0.33 3.00±0.58 9 3.25±0.25ab 4.00±0.58 3.00±0.58 3.33±0.33 2.67±0.33 12 2.25±0.18c 2.67±0.33 2.00±0.11 2.33±0.33 2.00±0.58 15 3.42±0.19a 4.00±0.13 3.33±0.33 3.67±0.33 2.67±0.33 18 3.50±0.26a 4.33±0.33 2.33±0.33 3.67±0.33 2.67±0.67 0

3.63±0.21A 2.75±0.18bC 3.08±0.16B 2.50±0.16C Means (T) <0.0001 P

0.00

02

1.83±0.21bc 2.33±0.33 1.33±0.33 2±0.58 1.67±0.33 0 Large follicles (9-12 mm)

2.5±0.15a 2.67±0.33 2.33±0.33 2.67±0.33 2.33±0.33 3 1.67±0.14c 2.00±0.14 1.67±0.33 1.67±0.33 1.33±0.33 6 2.83±0.21a 3.33±0.33 2.67±0.33 3±0.58 2.33±0.33 9 2.67±0.28a 3.67±0.33 2.33±0.33 2.67±0.67 2.00±.58 12 2.33±0.22ab 3.00±0.58 2.00±0.58 2.33±0.33 2.00±0.14 15 2.33±0.19ab 2.67±0.33 2.00±0.58 2.67±0.33 2.00±0.11 18 2.67±0.08a 3.33±0.33 2.33±0.33 2.67±0.33 2.33±0.33 0

2.88±0.15A 2.08±0.15bC 2.46±0.16B 2.00±0.12C Means (T) <0.0001 P

<0.0

001

2.75±0.18b 3.33±0.33 2.67±0.33 2.67±0.33 2.33±0.33 0 Ovulatory follicles (>12 mm)

1.75±0.13d 2.00±0.14 1.67±0.33 2.00±0.11 1.33±0.33 3 1.92±0.19d 2.33±0.33 1.67±0.33 2.00±0.58 1.67±0.33 6 1.25±0.13e 1.67±0.33 1.00±0.10 1.33±0.33 1.00±0.10 9 2.00±0.17cd 2.67±0.33 1.67±0.33 2.00±0.10 1.67±0.33 12 1.83±0.17d 2.33±0.58 1.67±0.33 2.00±0.10 1.33±0.33 15 2.42±0.15bc 2.33±0.33 2.33±0.33 2.67±0.33 2.33±0.33 18 3.33±0.28a 4.33±0.33 3.00±0.58 3.67±0.33 2.33±0.33 0

2.63±0.19A 1.96±0.16C 2.29±0.16B 1.75±0.14C Means (T) <0.0001 P

Values in the same row with different superscripts (A, B, C) are different (P<0.05) Values in the same column with different superscripts (a, b, c, d, e) are different (P<0.05) C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate


66

Fig. (16): Effect of treated cows with antioxidant on number of different size of ovarian follicles of Aberdeen Angus cows during estrus period

Research results in beef cattle indicate that primiparous cows fed

complexed trace minerals had confirmed pregnancies 10 days earlier (Swenson,

1998), 17 to 35% improvement in artificial insemination conception rates

(Ansotegui et al., 1994) and an increase in number of ova ovulated per heifer

(6.3 versus 2.8). Manspeaker et al. (1987) found that supplementation of dairy

heifers with Cu, Zn, Mn, Fe and Mg (chelated form) exhibited higher number of

mature follicles 30–80 days post-partum compared if compared to non-

supplemented (35 vs. 20%, respectively). Swenson, (1998) supplemented Cu,

Zn, Co and Mn as either inorganic sulfate or in a complexed form to first-calf

heifers showed that even though the percentage of significant structures

(follicles greater than 12 mm and/or corpora lutea as determined by rectal

palpation) and cows exhibiting oestrus by day 45 was lower when complexed

minerals were supplemented, the percentage of cows bred by artificial insemination was improved. Recent studies by González-Maldonado et al.

(2017) cocluded that there was no significant effect of injected trace minerals on

follicular and corpus luteum development in cows .

https://www.ncbi.nlm.nih.gov/pubmed/?term=Gonz%C3%A1lez-Maldonado%20J%5BAuthor%5D&cauthor=true&cauthor_uid=28385394


67

Regarding the effect of Zn supplementation on ovulation rate, the groups

that received 150 and 100 ppm ZnO had significantly (P<0.05) higher ovulation

rates than those in control (Abdel Monem and El-Shahat, 2011)

In terms of the percentage of ewes that ovulated and that had a functional

corpus luteum, the group supplemented with 150 ppm ZnO group had

significantly higher values (100%, P<0.05) than control, 50 ppm and 100 ppm

ZnO groups (50.0, 50.0, 75.0% respectively) (Abdel Monem and El-Shahat,

2011)

Vitamin E is necessary for normal reproduction in cattle (Allison and

Laven, 2000) and required for follicle and corpus luteum development (Das and

Chowdhury, 1999). The effect of vitamin E on fertility is mediated by a direct

antioxidant effect on the follicle (Al-Enazi, 2007). This is noteworthy, since

heat stress impairs fertility by follicle and oocyte disruption (Roth, 2008),

probably through inducing oxidative damage ( Roth, 2016 ) .

Data in Table (13) showed the significantly (P<0.05) highest size of

largest follicles in T3 then T1 and T2 compared with control one. Table (13): Effect of treated cows with antioxidant on size of largest follicle of Aberdeen Angus cows during estrus period (mean ± SEM)

P Means of days

Treatments (T) Days T3 T2 T1 C

<0.0

001

18.17±0.3a 19.20±0.12 18.3±0.06 17.6±1.05 17.57±0.15 0 13.03±0.18f 13.90±0.10 12.77±0.03 13.13±0.09 12.30±0.06 3 15.38±0.1d 15.80±0.06 15.13±0.12 15.57±0.03 15.03±0.12 6 12.83±0.09f 13.30±0.06 12.67±0.03 12.83±0.03 12.50±0.06 9 15.87±0.06c 16.18±0.02 15.73±0.03 15.9±0.06 15.67±0.03 12 14.27±0.08e 14.50±0.06 14.2±0.06 14.47±0.03 13.90±0.10 15 17.03±0.11b 17.5±0.02 16.87±0.09 17.20±0.06 16.53±0.03 18 18.26±0.16a 19.13±0.03 17.93±0.03 18.23±0.03 17.73±0.03 0 16.19±0.44A 15.45±0.42B 15.62±0.41B 15.15±0.41C Means(T)

<0.0001 P Values in the same row with different superscripts (A, B, C) are different (P<0.05) Values in the same column with different superscripts (a, b, c, d, e, f, g, h) are different (P<0.05) C: Control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3:

injected with vitamin E and selenium and supplementation with zinc sulphate


68

Abdel Monem and El-Shahat, (2011) found that supplementation of 100

to150 ppm zinc oxide to the basal diet of Baladi ewes significantly improved the

reproductive performance by increasing the number of large follicles and

ovulation rates. Furthermore, Abdel Monem and El-Shahat, (2011) indicated

that A significantly (P<0.05) higher population of large follicles (2.5±0.5 and

3.0±0.6 follicles) were found in ewes supplemented with 100 and 150 ppm ZnO

than in control and 50 ppm ZnO groups (1.0±0.2 and 1.0±0.3 follicles,

respectively).

Regarding to vitamin E, there is little information about its effect on corpus luteum functionality. Vierk et al. (1998) demonstrated that vitamin E

supplementation protects the corpus luteum from apoptosis.

Data in table (14) showed the significantly highest size of corpus luteum

in T3 then T1 and T2 compared with control one.

Table (14): Effect of treated cows with antioxidant on size of corpus lutem of Aberdeen Angus cows during estrus period (mean±SEM)

Values in the same row with different superscripts (A, B, C, D) are different (P<0.05) Values in the same column with different superscripts (a, b, c, d, e, f, g, h) are different (P<0.05) C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

Recent research concluded that injections of vitamin E did not affect

either the development of the preovulatory follicle and corpus luteum or

pregnancy rates in Holstein dairy cattle under heat stress conditions ( Gonzalez et al., 2017) .

P Means of days


<0.0

001

8.16±0.15h 8.85±0.04 8.01±0.06 8.28±0.05 7.51±0.15 0 10±0.13f 10.60±0.04 9.86±0.08 10.11±0.08 9.45±0.13 3

14.83±0.15e 15.50±0.10 14.53±0.03 15.07±0.09 14.24±0.14 6 18.18±0.18c 19.03±0.19 17.67±0.13 18.40±0.12 17.63±0.12 9 24.62±0.6a 27.50±0.40 23.87±0.12 24.90±0.38 22.20±0.31 12 19.87±0.17b 20.47±0.18 19.60±0.12 20.27±0.07 19.13±0.03 15 16.14±0.09d 16.50±0.15 16.00±0.10 16.23±0.03 15.83±0.03 18 9.48±0.18g 10.28±0.13 9.11±0.11 9.77±0.01 8.77±0.18 0

16.09±1.22A 14.83±1.09C 15.62±1.13B 14.34±1.04D Means (T) <0.0001 P


69

4-5-6-2. Concentration of estrogen and progesterone hormones

Data in Table (15) and Figure (17) showed a significant increase of

estrogen hormone level in Aberdeen Angus cows treated with antioxidant

compared to control group. Table. (15): Effect of treated cows with antioxidant on estrogen concentrations (pg/ml) during estrous period of Aberdeen Angus cows (mean±SEM)

P Means of days Treatments (T) Day T3 T2 T1 C

< 0.

0001

21.15±0.27a 22.01±0.11 21.28±0.07 21.57±0.07 19.73±0.26 0 11.06±0.21g 12.03±0.09 10.71±0.11 11.28±0.12 10.23±0.04 3 14.59±0.14e 15.22±0.15 14.34±0.03 14.69±0.09 14.08±0.09 6 10.85±0.08h 11.21±0.04 10.74±0.02 10.93±0.03 10.53±0.05 9 15.47±0.15d 16.23±0.04 15.23±0.03 15.51±0.1 14.91±0.03 12 12.61±0.13f 13.19±0.13 12.34±0.06 12.81±0.02 12.08±0.11 15 17.65±0.14c 18.21±0.05 17.32±0.02 17.96±0.02 17.10±0.14 18 20.97±0.2b 21.87±0.03 20.67±0.02 21.26±0.04 20.07±0.11 0

16.25±0.82A 15.33±0.81C 15.75±0.81B 14.84±0.75D Means(T) < 0.0001 P

Values in the same row with different superscripts (A, B, C, D) are different (P<0.05) Values in the same column with different superscripts (a, b, c, d, e, f, g, h) are different (P<0.05) C: control, T1:injected with vitamin E and selenium ; T2: supplementation with zinc sulphate T3: injected with vitamin E and selenium and supplementation with zinc sulphate

Fig. (17) Effect of treated cows with antioxidant on estrogen concentrations (pg/ml) during estrous period of Aberdeen Angus cows


70

The highest levels of estrogen hormone were recorded in T3 group

followed by T1 and T2 in comparison with control one. This result agree with

Ali et al, (2017) who reported that significant increase in estrogen levels in dairy

cows that injected with vitamin E and selenium compered to control .

Zinc has a key role in the physiology of the reproductive system (Hafiez

et al., 1990). It is known that zinc deficiency particularly in the diet leads to

hypogonadism (Nishi, 1996). Om and Chun, (1996) showed that zinc

deficiency led to an inhibition in LH and estrogen levels. An inspiring clue of

the relation between zinc and female reproductive system is that estradiol and

progesterone receptors obtained from calf uterus were bound to iminodiacetate-

sepharose chelate colons that contained zinc (Vallee and Falchuk, 1993).

Moreover, zinc was a significant stimulator in estradiol synthesis is a remarkable finding supporting the relation found between zinc and estrogen (Humeny et

al.,1999).

Data in Table (16) and Figure (18) showed a significant increase of

progesterone hormone level in Aberdeen Angus caws treated with antioxidant

compared with control group. Table (16): Effect of treated cows with antioxidant on progesterone concentrations (ng/ml) during estrous period of Aberdeen Angus cows (mean±SEM)

P Means of days


< 0.

0001

0.59±0.02g 0.67±0.03 0.57±0.01 0.61±0.02 0.50±0.02 0 0.83±0.01f 0.88±0.02 0.81±0.01 0.84±0.01 0.80±0.01 3 2.48±0.11e 2.93±0.09 2.23±0.03 2.67±0.13 2.07±0.15 6 7.44±0.11c 8.01±0.11 7.23±0.04 7.44±0.10 7.07±0.08 9 13.11±0.14a 13.71±0.06 12.99±0.06 13.28±0.12 12.48±0.19 12

9.70±0.1b 10.00±0.15 9.57±0.09 9.93±0.09 9.30±0.06 15 3.67±0.08d 4.00±0.10 3.47±0.07 3.80±0.06 3.40±0.06 18 0.77±0.02f 0.86±0.01 0.72±0.01 0.78±0.01 0.71±0.01 0

5.13±0.96A 4.7±0.92C 4.92±0.93B 4.54±0.89D Means(T) < 0.0001 P

Values in the same row with different superscripts (A, B, C, D) are different (P<0.05) Values in the same column with different superscripts (a, b, c, d, e, f, g, h) are different (P<0.05) C: control, T1:injected with vitamin E and selenium ; T2: Supplementation with zinc sulphate T3: injected with vitamin E and selenium and Supplementation with zinc sulphate


71

Bohr and Dial, (1982) suggested that the lower progesterone in hot

months might be due to indirect effect of increase ambient temperature that lead

to fluctuation in LH level, which has primary stimulation effect on progesterone

(P4) secretion in domestic animal. In the present study the averages of

progesterone hormone levels in Aberdeen Angus cows treated with antioxidant

agents showed significantly increased compared with control group. These results are in agreement with Ali et al. (2017) who found that vitamin E and

selenium supplementation raised plasma progesterone levels in Holstein cows. Moreover, Yildiz et al. (2015) found that that there was a significant increase in

progesterone in dairy cows that injected with vitamin E and selenium compered

to control.

Fig. (18) : Effect of treated cows with antioxidant on progesterone concentrations (ng/ml) during estrous cycle of Aberdeen Angus cows

Copper and zinc play an important role in regulating progesterone

production by luteal cells via involvement of superoxide dismutase (Sales et al.,

2011). Zinc is involved in the reorganization of ovarian follicles which are the

source of progesterone. This occurs through the involvement of

metalloproteinase-2 (MMP-2) enzyme, which is a member of zinc


72

endopeptidase family Gottsch et al. (2000). Positive correlation was reported

between serum progesterone level and copper-zinc in cows (Yildiz and Akar,

2001) .

Fig. (19): Estrogen and progesterone concentrations during estrous period of Aberdeen Angus cow

Estrogen and progesterone levels change continuously during estrous

cycle and these changes bring about changes in the hypothalamo-pituitary-

ovarian relationship that is the basis for endocrine regulation of reproductive

cycles in mammalian females (Pineda and Dooley, 2003). The progesterone

and estrogen concentrations during estrous cycle of Aberdeen Angus cows are

presented in Table (15,16) and Figure (17,18,19) . The overall mean

concentration of estrogen in the Aberdeen Angus cows during estrous cycle on

the day of estrus (day 0) and the day 3, 6, 9, 12, 15, 18, of estrous cycle and

subsequent estrus (day 0) were 21.15±0.27, 11.06±0.21, 14.59±0.14,

10.85±0.08, 15.47±0.15, 12.61±0.13, 17.65±0.14 and 20.97±0.2 pg/ml,

respectively. Similar to the concentration of estrogen in the present study, Naik

et al. (2013) in Punganur cows, Shukla et al. (2000) in crossbred cows, Coe

and Aldrich, (1985), Jimenez, (1988) , Gupta et al. (1998) and Selvaraju et


73

al. (2002) also recorded the same concentrations at different days of estrous

cycle. On the other hand, higher concentration of estrogen on different days of estrous cycle than the present finding were recorded by Mehrotra et al. (2005)

in Holstein Friesian x Hariana cattle, Mutha Rao et al. (2005) in Ongole cows,

Singh et al. (2006) in Sahiwal crossbred heifers. Estrogen concentrations in the

present study were higher than the reports of Agarwal et al. (1989) in cows,

Purohit et al. (2000) and Venkatesan et al. (2005) in indigenous cows,

Ecthernkamp and Hansel, (1973), and Glencross and Pope, (1981) in exotic

cows who recorded lower concentrations of estrogen on different days of estrous

cycle when compared to that in Punganur cattle. These inconsistencies in the

concentration of estrogen hormone might be due to the differences in the

sampling frequency, seasons and climate at the time of sampling and age and physiological stage (Lactating or not) of the animals (Alvarez et al., 2000).

Estrogen concentration showed a significant (P<0.01) decrease from day

‗0‘ to day 3 of estrous cycle which might be due to the ovulation and subsequent

development of luteal tissue under the influence of luteinizing hormone (Hafez,

2008 and Noakes et al., 2001). Later, the concentration of estrogen from day 3

to day 6 showed a significant (P<0.05) increase but the concentration of

estrogen did not reach the peak levels which indicated the development of first dominant follicle (Alvarez et al., 2000). Immediately after day ‗6‘ the

concentration of estrogen decreased significantly indicating atresia of dominant

follicle developed during first follicular wave (Hafez, 2008 and Noakes et al.,

2001). Later, the concentration from day 9 onwards estrogen showed an

increasing trend up to subsequent estrus day ‗00‘ at which time the

concentration of estrogen was at peak level which might be due to the development of preovulatory follicle (Alveraz et al., 2000 and Hafez, 2008).

The overall mean concentration of progesterone in the Aberdeen Angus

cows during estrous cycle on the day of estrus (day 0) and day 3, 6, 9, 12, 15,


74

18, of estrous cycle and subsequent estrus (day 0) were 0.59±0.02, 0.83±0.01,

2.48±0.11 , 7.44±0.11, 13.11±0.14, 9.7±0.1, 3.67±0.08, and 0.77±0.02 mg per

ml, respectively. The concentration of progesterone in the present study gained support from the findings of Echterkamp and Hansel, (1973) , Mutha Rao et

al., (2005), Singh et al., (2006), Naik et al. (2013) they also recorded similar

concentrations of progesterone at different days of estrous cycle. But the concentrations of progesterone reported by Rabiee et al., (2002) at day ‗0‘ and

Yildiz et al., (2005) at different days of estrous cycle were higher than the

present findings. While, progesterone concentrations in the present study were higher than the reports of Shukla et al. (2000), Selvaraju et al. (2002), Mandal

and Prakash, (2003), Mehrotra et al. (2005), and Venkatesan et al. (2005)

recorded lower concentrations of progesterone on different days of estrous cycle

as compared to the concentration of progesterone in Aberdeen Angus cattle.

The progesterone concentration showed significant increase from day ‗0‘

to day 15. Later, from day 18 onwards, the progesterone concentration showed a

significant decreasing trend and finally came down to the lowest concentration.

This constant rise in the concentration up to day 18 might be due to the

functional activity of corpus luteum and the decreasing trend from day 18 might

be due to the initiation of luteolysis (Hafez, 2008). Furthermore, this study also

recorded the lowest progesterone concentrations on the day of estrus and this

gained support from the findings of all the authors who also recorded either

similar or higher or lower concentrations on the other days of estrous cycle than

the present study. These inconsistencies in the concentration of progesterone

hormone might be due to the differences in the sampling frequency, seasons and

climate at the time of sampling, age and physiological stage (lactating or non-lactating) of the animals (Alvarez et al., 2000).


75

4-5-6-3. Correlation between ovarian follicular and luteal development and

hormone profile

In this study, the increase in follicular size was associated with an

increase in estrogen concentration. Also, the increase in corpus luteum size was

associated with an increase in progesterone concentration.

Fig. (20) : A positive correlation between progesterone concentrations and corpus luteum during estrous period of Aberdeen Angus cows

The corpus luteum is a temporary endocrine structure in female mammals

that is involved in the production of relatively high levels of progesterone.

Fig. (21) : Ultrasonographic view from ovaries with non-vacuolated CL


76

Fig. (22) : Ultrasonographic view from ovaries with vacuolated CL

Fig. (23) : A positive correlation between estrogen concentrations and largest follicle during estrous period of Aberdeen Angus cows

Perry et al. (2014) found that a significant correlation between the

preovulatory follicle diameter and peak concentrations of estradiol was detected, regardless of suckling status. This relationship is similar to a report (Jinks et al.,

2013) of a significant correlation between follicle diameter at second GnRH and

concentrations of estradiol at second GnRH in cows not exhibiting standing

estrus.


77

Earlier researchers have reported a strong correlation between diameter of

the dominant follicle and estradiol concentrations in the follicular fluid during

the preovulatory period (Ireland and Roche, 1982 : Kruip and Dieleman,

1985)

Fig. (24) : Ultrasonographic view from ovaries with small follicle (< 5 mm)

Fig. (25) : Ultrasonographic view from ovaries with follicle ( 5-12 mm)

Fig. (26) : Ultrasonographic view from ovaries with Ovulatory follicle ( > 12 mm)

SUMMARY AND

CONCLUSION

Summary and Conclusion

78

5- SUMMARY AND CONCLUSION

This experiment was carried out at the Animal Production Experimental

farm, Faculty of Agriculture, New Valley University in cooperation with Animal

production Department, Faculty of Agriculture, Assiut University. The study

was started from July to September 2016 in the new valley governorate during

summer season.

A total number of sixteen healthy Aberdeen Angus cows about 3-4 years

of age and an average body weight of 460-520 kg were used in this study . The

animals were divided randomly in to four equal (4 animals each group) .

(Control ( C ) , vitamin E and selenium (T1) , zinc sulphate (T2) , and vitamin E

and selenium plus zinc sulphate (T3). The present study was aimed at

elucidating the effects of zinc sulfate administration and vitamin E + selenium

(Se), as a antioxidant agents, on thermoregulatory responses, hemato-

biochemical parameters and reproductive performance of Aberdeen Angus

cows under New Valley Conditions.

The obtained results could be summarized as the following :- 1-Ambient temperature, relative humidity and temperature-humidity

index (THI) during experimental period :- x The average ambient temperature during the experimental period ranged

from 38.3 o C to 45o C at 02:00 pm and 25.2 o C to 30.3 o C at 10:00 pm .

While, the respective average RH were 10% and 25%.

x The average values of THI were between 79.43 to 85.46 at 02:00 pm and

69.04 to 74.14 at 10:00 pm during the experimental period .

2-Effects of using antioxidant agents on thermoregulatory responses of

experimental cows :- Cows treated with antioxidant led to significant decrease in rectal

temperature, skin temperature and ear temperature, but the differences were not


79

significant . While, a significant decrease in respiration rate, pulse rate and hear

temperature was showed in of experimental animal .

3-Effects of using antioxidant agents on hematological parameters in experimental caw .

x White blood cells and it fraction :

Results indicated that the average number of white blood cell

(WBC) and Lymphocytes (LYM) of cows injected with vitamin E and

selenium (T1) was significant (P<0.05) higher than other treatments.

x Red blood cells, Hb and HCT

The results showed that no significant differences were found among

treatment in , red blood cell (RBC) count.

- Treated cows with T1 had higher (P<0.05) concentration of

hemoglobin (Hb), hematocrit (HCT), red Blood Cell Distribution

Width (RDW), and mean corpuscular volume (MCV) than other

treated cows

- The results revealed that mean corpuscular hemoglobin (MCH) and

mean corpuscular hemoglobin concentration (MCHC) increased

(P<0.05) for T1, T3 and T2, respectively compared to control.

4- Effects of using antioxidant agents on biochemical metabolites

parameters in experimental cows:

x Serum concentrations of total protein, globulin, glucose, alanine

aminotransferase ( ALT), Urea nitrogen, and creatinine did not differ

significantly due to treatment. Moreover, serum cholesterol, albumin and

aspartate aminotransferase (AST) was significantly higher (p < 0.05) in

T1 group than other treatments. In addition, the highest total antioxidant

capacity (TAC) did not differ significantly due to treatment.

x Cows treated with antioxidant agent showed higher (P<0.05)

concentration of serum albumin .


80

x Serum concentration of cholesterol was significantly (P<0.05) higher in

cows injected with vitamin E and selenium (T1) than the other treatment .

x Serum concentration aspartate aminotransferase (AST) was significantly

(P<0.05) higher in cows injected with vitamin E and selenium (T1) than

other groups . While cows supplemented with zinc sulphate (T2) had a

significant decrease (P< 0.05) in aspartate aminotransferase (AST)

compared with other groups

x Serum concentration of alanine aminotransferase ( ALT) did not differ

significantly due to treatment .

x The highest (P< 0.05) total antioxidant capacity (TAC) was recorded for

cow groups treated with zinc sulphate and vitamin E + Se (T3 ) followed

by those groups treated with vitamin E + Se (T1) , then zinc sulphate (T2

) and to lesser extent of control treatment .

5-Effects of using antioxidant agents on reproductive performance in

experimental cow.

x Response to Synchronization

The present study illustrated that purebred cows exhibited no difference in

their response to synchronization due to treatments.

x Ovarian follicles

The highest number of all different size of ovarian follicles were recorded

for cows treated with zinc sulphate and vitamin E + selenium (T3 )

followed by those groups treated with vitamin E + selenium (T1) , then zinc

sulphate (T2 ) and to lesser extent of control treatment .

-The largest follicles and corpus luteum were recorded for cow in (T3 )

groups followed by (T1) , then (T2 ) and to lesser extent of control.

x First service conception rate

Cows treated with vitamin E + Se (T1), zinc sulphate (T2) or there

combination (T3) showed a significant (P<0.05) increase in the pregnancy

rate at first service than control group


81

x Number of services per conception

Cow treated with zinc sulphate and vitamin E + Se (T3 ) , vitamin E + Se

(T1) , zinc sulphate (T2 ) showed a significant (P<0.05) decrease the

number of services per conception compared with control .

x Conception rate

The highest conception rate ( 100%) was recorded for cow treated with

zinc sulphate and vitamin E + Se (T3 ) , vitamin E + Se (T1) , and zinc

sulphate (T2 ). While, lesser conception rate (75%) extent of control

treatment.

x Size of embryo and Embryonic vesicle

The largest size of embryo and amniotic vesicle were recorded for cows

treated with zinc sulphate and vitamin E + Se (T3 ) followed by those

groups treated with vitamin E + Se (T1) , then zinc sulphate (T2 ) and to

lesser size extent of control group .

x Concentration of estrogen and progesterone hormone

The highest levels of estrogen and progesterone hormone were recorded

in T3 group followed by T1 and T2 compared with control one.

Conclucion

It was concluded that the antioxidant agents which used in the present study

such as vitamin E + selenium and zinc sulphate in Aberdeen Angus cows have

an ameliorative effect on their physiological responses including thermo-

respiratory parameter and improve the reproductive performance, without any

harmful effect on animal health , blood hematology and serum biochemical

indices. Under heat stress of new valley conditions

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.

ARABIC SUMMARY

انهخص انعرب

1


حأثر اسخخذاو يىاد يضاداث الأكسذة عه الأداء انخاسه وانخظى انحراري لأبقار

الأبرد أجس ححج ظروف انىادي انجذذ

، اندذذ اناد فشع انضساػت ، كهت انسا، الإخبج يضسػت قغى ف انخدشبت ز جأخش

ي انذساعت بذأث قذ أعغ. خبيؼت ، انضساػت كهت ، انسا الإخبج قغى يغ ببنخؼب أعغ خبيؼت

.انصف فصم خلال اندذذ اناد يسبفظت ف 6116 عبخبش إن ن شش

ي اندغى ص يؼذل عاث 4-3ف ػش بقشة أبشد أدظ 16 انذساعت ز ف اعخخذو

(. يب نكم زابث 4) يخغبت يدػبث أسبغ إن ػشائب انسابث حقغى حى. كدى 461-561

انغهو E فخبي ،( T2) انضك كبشخبث ،( T1) انغهو E فخبي ،( C) كخشل)

حج انز الأعبع انغزائ انظبو ػه الأبقبس خغ حغزت حى((. T3) انضك بشخبثك إن ببلإظبفت

٪ 41 انقر حب ي خك الأعبع انغزائ انظبو. انهسى نبشت NRC(2000) نـ فقب صبغخ

أ د أعبع غزائ ببنظبو( C) الأن اندػت ف الأبقبس حغزت حى٪. 61 انؼهف انشكض

يهدى نكم زا كم 15انغهو بدشػت Eبفخبي (T1)حى زق اندػت انثبت بب لاثيك

يهدى / بقشة يب 611 انضك نهؼهقت بؼذل حى اظبفت كبشخبث( T2) انثبنثت اندػت . اعبػ

انضك كبشخبث اظبفت أعبػ انغهو كم Eبفخبي زقب حى( T3) انشابؼت اندػت بب

يهدى / بقشة يب.611 نهؼهقت بؼذل

:انخان انحى عهى عهها انحصىل حى انخ انخائج حهخص ك

خلال (THI) ويؤشر انعبء انحراري انسبت انرطىبت , انجىت انحرارة درجت - 1

-: انخجربت انفخرة

دسخت 45 إن يئت دسخت 3..3 ي انخدشبت شةانفخ خلال انسطت انسشاسة دسخت يخعػ حشاذ •

11011 انغبػت ف يئت دسخت 31.3 إن يئت دسخت 65.6 انظش بؼذ 6011 انغبػت ف يئت

٪. 65 ٪ 11 انقببم RH يخعػ انشغبت انغبت كب ، ز ف. يغبء

ف 34.14 نیإ 64.14 ء عبي 16011 نعبػتا ف 5.46. نیإ 34.43 ب يب THI وق غعيح ثبنغ •

.بتسنحخا سةنفحا لخلا ء يعب 11011 نعبػتا


2

: لأبقار الابرد اجس انخظى انحراري عهى الأكسذة يضاداث اسخخذاو حأثر - 2

زشاسة دسخت ف غش يؼ اخفبض اظشث الأكغذة بعبداث انؼبيهت الأبقبس

يؼ ف اخفبض أظشث ز ف. لأرا زشاسة دسخت اندهذ زشاسة دسخت انغخقى

.انخدشبت انسابث ف انشؼش زشاسة دسخت انبط يؼذل انخفظ، يؼذل

ف ابقار الابرد اجس انؤشراث انبىنىجت عهى الأكسذة يضاداث اسخخذاو حأثر – 3

:ويكىاحها انبضاء انذو خلاا • LYM)انخلاب انهفبت ) WBC و انبعبءػذد كشاث انذ يخعػ أ إن انخبئح أشبسث

.الأخش انؼبيلاث ي أػه( P <0.05) يؼب كب( T1) انغهو E بفخبي انسقت نلأبقبس

انحراء انذو خلاا •

خلاب انذو انسشاء. ف ػذد انؼبيلاث ب يؼت فشق حخذ لا أ انخبئح أظشث

(Hb)انخهب ف ( P <0.05) حشكضا أػه T1دػت انثبت ف ان أبقبس كبج -

انسدى انخقشب نخهت انذو انسشاء RDW)حصغ كشاث انذو انسشاء ) ((HCTانبحكشج

((MCV ف اندػت انشابؼت انخدة حهك ي T3 الأن C انثبنثتT2

(MCHC) حشكض انخهب MCH)) خهبيسخ انخهت ي ان أ انخبئح أظسج -

.بدػت انغطشة يقبست انخان ػه T1 T3 T2 ف اندػبث ( P <0.05) صادحب

ف ابقار الابرد انبىكائت الأض يعايلاث عهى الأكسذة يضاداث اسخخذاو حأثر -4

اجس

x انؼبيهت أظشث (T1 )انكنغخشل، الانبي ي نغشوا حشكضاث ف يؼت صبدة

ف يؼت اخخلافبث حخذ لا بب. انؼبيلاث بببق يقبست (AST) إضى شبغ يغخ

يغخ انكشبح، انسب، اندهكص، اندهبن، انكه انبشح ي انغشو حشكضاث

. انؼبيلاث ب (ALT) إضى شبغ

x أػه حشكضا الأكغذة بعبداث نؼبيهتا الأبقبس أظشث (P <0.05 )بب ي ، لا الأنبي

انكه انبشح ي انغشو حشكضاث ف يؼت اخخلافبث حخذ اندهبن


3

x أػه يؼب انغشو ف انكنغخشل حشكض كب (P <0.05 )بفخبي انسقت الأبقبس ف E

.ؼبيلاث الاخش ببن يقبست( T1) انغهو

x انصم حشكض كب (AST) يهسظ بشكم (P <0.05 )بفخبي انسقت الأبقبس ف أػه E

انضك بكبشخبث انكهت الأبقبس أ ز ف. الأخش ببندػبث يقبست( T1) انغهو

(T2 )يؼب اخفبظب عدهج (P <0.05 )ف AST الأخش ببندػبث يقبست.

x حشكض هفخخ نى ALT)) انغشو بشكم .انؼبيلاث بغبب كبش ف

x انكهت الأكغذة نعبداث حشكض اػه انخبئح أظشث (TAC )انشابؼت اندػت ف T3) )ثى

.حشكض اقم عدهج انخ انعببطت ببندػت يقبست( T2) انثبنثت ثى( T1) انثبت اندػت

ابقار الابرد اجس. ف انخاسه داءالأ عهى الأكسذة يضاداث اسخخذاو آثار - 5

الاسخجابت نخظى انشاع • اعخدببخب ف اخخلافبث ف اعخدببت الابقبس ف كم اندػبث خد ػذو أظشث انخبئح

% 111انشبع ، كبج غبت الاعخدببت نخظى انشبع ف كم الابقبس نخضاي

انبضه انحىصلاث • حى بؼبيهخب انخ ف الابقبس انبع انسصلاث ازدبو ف خغ ػذد أكبش حغدم حى

+ E بفخبي انؼبيهت اندػبث حهك حهب( T3) انغهو+ E يغ انسق بفخبي انضك بكبشخبث

يدػ انكخشل ، كب ػذد أقم صلا ان( T2) انضك كبشخبث ثى ،( T1) انغهو حغدم حى ف

اندغى الاصفش زدى أكبش اندػ نهسصلاث انبع + E فخبي انضك بكبشخبث انؼبيهت ف

اندػت انؼبيهت ثى ،( T1) انغهو+ E بفخبي اندػ انؼبيهت حهك حهب( T3)انغهو

اصغشى زدب ظش ف يدػت انكخشل ( T2) انضك بكبشخبث

نحم ي اول حهقحيعذل ا • أ انضك بكبشخبث انؼبيهت الأبقبس أظشث يؼت صبدة ا الاث يؼب + انغهو Eفخبي

ي ال حهقس انسم يؼذل ف

عذد انخهقحاث نحذود انحم • ، (T3) انغهو+ E فخبي انضك بكبشخبث انؼبيهت الابقبس يدػبث أ انخبئح أظسج

ادث ان حقهم ( T2) انضك انؼبيهت بكبشخبث ،( T1) انغهو + E بفخبي انؼبندت اندػبث

.ػذد انخهقسبث انلاصيت نسذد انسم


4

انحم يعذل • E فخبي انضك بكبشخبث انؼبيهت الابقبس ندػبث٪( 111) نهسم يؼذل أػه حغدم حى

اندػبث انؼبيهت ،( T1) انغهو+ E بفخبي انؼبندت اندػبث ، (T3) انغهو +

. ٪( 35) ف يدػت انكخشل انسم بب كب يؼذل( T2) انضك بكبشخبث

انجت وانحىصهت انج حجى • ضكان بكبشخبث الابقبس انؼبيهت ندػبث اندت انسصهت نهد زدى أكبش حغدم حى

ثى ،( T1) انغهو + E بفخبي انؼبيهت اندػت حهك حهب( T3) انغهو + E فخبي

ب الأقم اخشا( T2) انضك اندػت انؼبيهت بكبشخبث انكخشل ف يدػت زد

انبروجسخرو الإسخروج و حركز هريى • يدػت الابقبس انؼبيهت ف بشخغخشان الاعخشخ نشي يغخبث أػه حغدم حى

انغهو+ E بفخبي انؼبيهت اندػت حهب( T3) انغهو+ E فخبي انضك بكبشخبث

(T1 ) انضك بكبشخبث انؼبيهت اندػت ((T2 بدػت انكخشل يقبست.

انخلاصت

+ E فخبي ) انسبنت انذساعت ف تانغخخذي الأكغذة يعبداث أ إن انذساعت خهصج

ػه حأثش نب (انضك كبشخبث انغهو رنك ف بب ظأد أبشد بقبسلأ انفغنختالاعخدببت إدبب

يؤششاث انسا صست ػه ظبس حأثش أ د ، انخبعه الأداء اعخدببخب نهخظى انسشاس حسغ

اندذذ ببناد ظشف الاخبد انسشاس حسج ت انذو انبنخت انبكبئ

جاهعت اسوط كلت الزراعت

قسن الاخاج الحواى

حأثير اسخخذام هواد هضاداث الأكسذة عل الأداء الخاسل والخظن الحراري لأبقار الأبردي أنجس

تحج ظروف الوادي الجذذ

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