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Birsa Agricultural University, Ranchi
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ThesisItem Open Access EFFECT OF ENDOSULFAN ON GONADAL ENZYMIC ACTIVIT AND RESIDUAL EFFECT IN BLACK BANGAL BUCKS(Birsa Agricultural University, 1988) BANDYOPADHYAY, ANAND GOPAL; Mukherjee, S.K.it is concluded that the regional distribution pattern for L-isomer and B-isomer may help to explain the difference in biochemical toxic potential of these forms of endosulfan. The early variation of some of the marker enzymes of testicular functions suggests the role of various enzymes investigated in this study in the testicular toxicity of endosulfan. Although, at present, it may be difficult to understand the mechanism of action by which endosulfan causes the testicular injury but the significant decrease in dose dependent activity of enzymes involved in the normal functions of spermatogenesis and fertilization indicates that endosulfan is injurious to testis and functional damage can occur at comparatively low doses. From the above findings in the present study with organochlorine pesticide "Endosulfan" it may be concluded that "Endosulfan" pre-exposed feed when fed to bucks were found to produce hazardous effects on gonadal enzymic activity leading to decreased reproductive efficiency of Black Bengal bucks.ThesisItem Open Access ANTIOXIDANT STATUS OF PIG DURING DIFFERENT STAGES OF REPRODUCTION AND ITS RELATION WITH REPRODUCTION STRESS(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2011) Mandi, Binod Bhushan; Prasad, R. L.Reproduction and developmental process accompany dynamic changes in metabolism and oxygen consumption. Therefore the byproducts are also generated in an extraordinary scale. Among these byproducts, reactive oxygen species (ROS) are more troublesome. Normally, there exist a balance between the oxidants and antioxidant. When the balance is disrupted towards an overabundance of ROS, oxidative stress occurs. This oxidative stress leads to many problems. Antioxidants vitamins are powerful and they are helpful in prevention of many reproductive disorders like Abortion, still birth, low birth weight, pre-eclampsia, premature birth, ROS induced fetal abnormalities, endometeritis, ovarian cancers, preterm labour. Instead of their important role in reproduction there are few trials investigating the antioxidant status during the reproductive stress. Pig being an important part of meat industry, a great 2 source of animal protein and a remunerative enterprise, especially in the tribal belt of Jharkhand. The present study was conducted on 8 T & D sows to compare the antioxidant vitamins status and their correlation with reproductive parameters during different physiological stages by estimating the concentration of antioxidant vitamins, biochemical parameters and some of the reproductive performance and the same have been summarized and concluded as follows: 1. There is significant (p < 0.01) increase in blood glucose level during early pregnancy, mid pregnancy, late pregnancy even after farrowing as compared to 0 day. 2. There is significant (p < 0.01) increase in total serum protein during estrus and pregnancy as well as after farrowing. 3. The total serum cholesterol level increased significantly during estrus but goes down throughout pregnancy and even after farrowing. 4. The level of β-carotene significantly decreased slowly from estrus (22.91±0.02 μg/100 ml) to early, mid and late pregnancy as well as after farrowing (19.40±0.04 μg/100 ml). 5. Similarly vitamin C decreased from 0 day (531.31±1.66 μg/100 ml) to pregnancy and reached still lower (482.25±1.85 μg/100 ml) after farrowing. 6. However, vitamin E increased significantly from 3.51±0.04 μmol/L (0 day) to 3.87±0.03 μmol/L (estrus) but, decreased significantly in pregnancy (4.44±0.03 to 3.99±0.04 μmol/L) as well as after farrowing to 2.79±0.03 μmol/L. 7. Lipid peroxidation (LPO) of erythrocytes increased significantly in estrus, pregnancy and after farrowing as compared to 0 day. 3 8. Not a single case of abortion occurred during the present experiment, while the overall percentage of stillbirth and low birth weight were found to be 5% and 6.66% respectively in T & D sows. 9. Both stillbirth and low birth weight had negatively significant (p < 0.05) correlation with β-carotene concentration during early pregnancy. 10. Vitamin E level showed positive significant (p < 0.01) correlation during estrus with low birth weight. 11. The vitamin C concentration had negative correlation with stillbirth and low birth weight throughout the pregnancy and even after farrowing. 12. There was no valid correlation between abortion and β-carotene, vitamin E and vitamin C respectively throughout the pregnancy and even after farrowing. 13. The level of lipid peroxidation of erythrocytes showed negatively significant (p < 0.01) correlation with β-carotene and vitamin C. Whereas, the level of lipid peroxidation of erythrocytes showed positive significant (p < 0.05) correlation with blood glucose.ThesisItem Open Access BIOCHEMICAL CHARACTERIZATION OF FMD VIRUS AND ITS IMPACT ON METABOLIC PROFILE, OXIDATIVE STATUS AND OVARIAN ACTIVITY IN CATTLE(Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2011) Nath, Rita; Prasad, Ram LakhanFoot and mouth disease virus (FMDV) is the first animal virus to be identified in 1898 by Loeffler and Frosch. It is known as one of the most fearful viral pathogens of animals since it is highly contagious among all species of cloven-hoofed animals including cattle, sheep, goats and swine . It is caused by a small single-stranded, positive- sense RNA virus of about 8500 nucleotides. It belongs to genus aphthovirus, family Picornaviridae, The term aphthovirus is derived from the Greek word 'aphtha', which refers to the vesicles found in the mouth and feet of affected animals. There are seven serotypes of FMD virus reported globally. Firstly the serotypes O, A and C were discovered and later on three additional serotypes found in Southern Africa and were named accordingly as Southern African Territories types SAT1, SAT2, SAT3 and Asia-1. Oxidative stress (OS) is an active field of research in Veterinary medicine and has been implicated in numerous disease processes . Oxidative stress may impair health in dairy cows both directly and indirectly. It a negative effect on animal health and productivity as well as it has been implicated as a major initiator of tissue damage . A limited number of conditions in ruminant medicine have been investigated in regard to the effects of oxidative stress (Bras, 2010) but not on FMD infected animals. With this in mind, it is the goal of this study to summarize the present knowledge of oxidative stress in ruminant physiology and production during FMD. However, the epidemiology and etiology of FMD have been extensively investigated (Yang et al., 1993) but, a few published reports on haematological and biochemical parameters of cattle with FMD are available (Elitok at el. 1999; Lubroth, 2002 and Gokce et al., 2004). Keeping these in view, the work was designed to run in cattle with the following objectives so that FMD in Indian origin is better understood and a more effective treatment can be given. i) Biochemical characterization of the FMD virus. ii) To study the metabolic profile of FMD infected cattlea. Haematological parameters b. Biochemical parameters iii) To study the impact of FMD on the oxidative status of infected cattle. iv) To study the impact of FMD on the ovarian activity of infected cattle. The present study was carried out in two groups of cross bred cattle. Group I consisted of ten (10) crossbred cattle from Instructional Bovine farm, Ranchi Veterinary College, that were clinically healthy and group II consisted of thirty (30) crossbred cattle having clinical signs of FMD. Animals without characteristic lesions of FMD were not considered for the study. After estimation and statistical analysis by ANOVA , students’ t-test and co-relation coefficient of various hematological and biochemical constituents, the findings can be summarized as follows: Hematology: The concentration of hemoglobin in control and was found to be 9.966 ± 0.344 % that had a decreasing trend to 9.220 ±0.337 %. The PCV was observed to be 27.899±0.603 % in the healthy animals while in the FMD affected animals it decreased to 26.464±0.588 % but the difference was non significant. The ESR value was recorded as 0 in both the groups. The WBC count in control group was recorded as 5.775±0.320 X 103 which slightly increased to 5.850±0.108 X103 in the test group while RBC count in group I was 5.588±0.220 X 106 which decreased significantly (P<0.05) to 5.065± 0.132 X 106 in group II. The red blood indices ie MCH, MCV and MCHC in control animals were found to be 17.915±0.693 pg, 50.283±2.145 fl and 35.795±1.363% and in the test group animals the values showed a non significant variation. Clinical Biochemistry: The concentration of protein in the healthy animals were recorded as 6.998 ± 0.288 gm/dl while in the FMD affected animals it decreased to 4.325±0.222 mg/dl while the serum albumin concentration in group I was 4.325±0.222 gm/dl and in group II was 3.230±0.251 gm/dl . Both total serum protein and albumin decreased highly significantly (P<0.01). In the healthy animals the serum globulin concentration was recorded to be 2.637±0.296 gm/dl while in FMD affected animals was 2.516±0.174 gm/dl .The A:G ratio was 1.777±0.201 and 1.334±0.163 in the control and FMD affected animal and both showed a non-significant variation.The concentration of serum cholesterol, urea, BUN, uric acid and calcium in group I was 116.792 ± 11.615, 15.637±0.9287, 7.128±0.413, 2.927±0.159 and 8.706±0.338 mg/dl respectively and in group II all decreased highly significant (P< 0.01) to 85.701±3.795, 12.075±0.430, 5.588 ± 0.125, 2.525±0.166 and 6.589±0.366 mg/dl respectively. Serum creatinine, BUN:Creatinine ratio and blood glucose concentration was estimated to be 1.297±0.267 mg/dl, 6.527 ± 1.212 and 45.769 ± 2.210 mg/dl respectively in the healthy animals and all showed a decreasing trend in the diseased cows. The total serum bilirubin, direct bilirubin (conjugated) and indirect bilirubin (non-conjugated) in the healthy animals were 0.467±0.055, 0.329±0.015 and 0.138±0.023 mg/dl respectively. In the FMD affected animals the concentrations changed slightly but indirect serum bilirubin increased significantly (P< 0.05) to 0.271±0.089 mg/dl. Enzymology: The alanine transaminase (ALT) activity in control group was observed to be 16.789±2.840 U/L and in test group it increased slightly while aspartate transaminase (AST) activity was 78.032±3.187 U/L in the control group which increased significantly(P< 0.05) to 95.105±5.582 U/L in the test group. Alkaline phosphatase (ALP) activities in serum in group I and II were almost same in both the groups ie 39.258±2.195 U/L and 39.184±3.402U/L. Superoxidase dismutase (SOD) activity in hemosylate of group I and II were 391.023±25.930 U/ml and 337.756±9.502 U/ml respectively. SOD activity showed a significant (P< 0.05) decrease in group II. Glutathione peroxidase (GSH-Px) and Glucose-6-phosphate dehydrogenase (G6PD) activity in hemoysate were observed to be 123.556±5.208 U/ mg Hg and 11.907±1.287 U/gm Hb respectively in group I which decreased slightly in group II. Vitamins: The concentration of β- carotene in plasma was 1.075 ± 0.134 mg/dl in control and in the test group it was almost same but ascorbic acid in healthy animals was 11.229±0.814 mg/dl and in FMD affected animals it decreased significantly (P<0.05) to 8.856±0.603 mg/dl . Lipid peroxidation: The amount of MDA produced was 1.904±0.132 nmol/gm Hb in the healthy animals which increased slightly to 2.287±0.371 nmol/gm Hb in the FMD affected animals. Hormone activity: The serum progesterone concentration during estrus phase in control group was 0.77± 0.11 ng/ml and during luteal phase the was 7.310±1.546 ng/ml which decreased slightly in the deseased animals both during estrus and luteal phase. Molecular weight and mobilities of proteins: The molecular weight of the FMD viral proteins I and II of trivalent A, O and Asia 1 FMD vaccine were 101 and 62 K Da respectively as per known protein molecular weight standard. Both in the control and FMD affected serum sample seven distinct protein bands were observed and were from the top of the gel were as ϒ globulin, β globulin, unidentified protein I, heptoglobin, albumin, α2 globulin and unidentified protein II in the control serum sample. Their molecular weights were 150, 140, 120, 100, 60, 48, and 33 K Da respectively as calculated from the semi log graph. In the FMD affected sample, δ globulin was undetectable but an extra band was noticed in the infected serum sample which was identified as the FMD viral protein and their molecular weight was 40 K Da. Correlation between the different parameters: It was observed that among the blood haematological parameters, a highly significant (P<0.01) positive correlation was observed between Hb and MCH and MCHC, between PCV and MCV, between MCH and MCV and MCHC whereas a highly negative significant (P<0.01) correlation was observed between RBC and MCV. Among the serum protein fractions and liver enzymes, a positive significant (P<0.05) correlation was observed between protein and albumin and A:G ratio, between ALT and ALP but a significant (P<0.05) negative correlation was observed between albumin and globulin and between globulin and A:G ratio. A highly positive significant (P<0.01) correlation was observed between albumin and A:G ratio and a highly negative significant (P<0.01) correlation was observed AST and ALT . Among the serum biochemical variables, a positive significant (P<0.05) correlation was observed between direct bilirubin and calcium and between between indirect bilirubin and glucose but a negative significant (P<0.05) correlation was observed between urea and glucose, between BUN and glucose and between direct bilirubin and cholesterol. A highly positive significant (P<0.01) correlation was observed between urea and BUN and between total bilirubin and indirect bilirubin and glucose . Among the oxidative stress biomarkers, a positive significant (P<0.05) correlation was observed between protein and albumin and SOD, between albumin and ascorbic acid and between MDA and ascorbic acid but a negative significant (P<0.05) correlation was observed between uric acid and ascorbic acid whereas between MDA and SOD, there was a highly significant (P<0.01) negative correlation. From the study it was concluded that: Hematological and biochemical values were an efficient tool for evaluation of physiological status and metabolic disorders which have a great relation to health status of the animal. The animals affected with Foot and Mouth Disease showed an alternation in the metabolic profile. There might be liver dysfunction as the liver enzyme activities (liver function test) were increased indicating affected the metabolic profile. Metabolic profile may also be affected also due to endocrinopathy, but to what extend it occurs in cattle with FMD requires further study. Thus, biochemical and hematological alternations may indicate the development of anemia, liver dysfunction and endocrinopathy. The disturbed oxidative status of the animals affected with FMD reflected that they were in oxidative stress condition. Most of the antioxidant biomarkers showed a significant change in the infected animal. The depletion in the body’s antioxidant resource have resulted from its action to counter balance the toxic effects of the oxidants which is produced in increased amount during FMD confirming the occurrance of oxidative stress during FMD. The present work summarizes the knowledge of oxidative stress in ruminant physiology and production during FMD and highlighting the need for continued research on oxidative stress in ruminant medicine since it is still in the early stage of development. The low serum progesterone concentration (non-significant) in FMD affected cow is associated with sub functional ovarian activity as these animals may have no corpora lutea which is the main source of serum progesterone. Oxidation stress as well as less feed intake has a significant role in cessation of ovarian activity by affecting the growth of graafian follicles and oocyte maturation. Thus, it can be concluded that during FMD, the ovary is also involved. The study has also provided information regarding the molecular weights and mobilities of the FMD viral proteins in the trivalent FMD vaccine and FMD affected serum sample. It was also seen that both the control and test serum sample segregated into seven distinct bands but in the FMD affected serum sample the protein ϒ – globulin was undetected but an additional FMD viral protein band was noticed. Thus, the FMD affected animals should be treated symptomatically as well as be given hepato stimulants. They should be provided with supplements rich in trace minerals (eg Fe, Cu, Zn, Co etc.) and antioxidant vitamins (eg Vit A, Vit C, Vit E) which would be beneficial for the antioxidant defence system. Again, further studies should be made to set the levels at which it would be beneficial for the antioxidant defence system. Finally, the present study may provide a better understanding of the disease process and clinical pathology of Foot and mouth disease for a better line of treatment.
