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20233
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Subject: Veterinary Physiology × End date: 2023 × Start date: 2023 ×
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    ThesisItemOpen Access
    GENE EXPRESSION PATTERNS OF SELECTIVE MONOCARBOXYLATE TRANSPORTERS IN BUBALINE SPERMATOZOA
    (SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA, 2023-10) NIRUPAMA .M.U.L; SRINIVASA PRASAD .CH (MAJOR); IQBAL HYDER; ASWANI KUMAR .K; MUTHA RAO .M
    The objective of the present study was to evaluate the gene expression of certain monocarboxylate transporters in bubaline spermatozoa. Eight adult healthy buffalo bulls between three to six years of age were randomly selected from ABC Semen Station, Veeravalli, Krishna District, Andhra Pradesh. The fresh semen samples were collected from eight buffalo bulls once in a week for four weeks by AV method. As a preliminary quality check, fresh semen samples (4 samples from each bull, n=32) were evaluated at the semen station for volume, concentration, mass motility, individual progressive motility, viability, HOST, acrosomal integrity and the mean±SE values were found to be 4.09±0.26mL, 1243.875±65.621 million/mL, 3.86±0.34 and 70.04±9.92%, 73.42±0.49%, 73.98±0.56% and 87.75±0.52% respectively. With respect to evaluation of gene expression of monocarboxylate transporters, the fresh semen samples were transported at 37°C. The samples were divided in to two groups. Fresh uncapacitated as control (4 samples from each bull n=32) and fresh capacitated (n=32). It was taken care that fresh semen samples had sperm concentration of 50-60 million spermatozoa. In vitro capacitation of fresh semen samples was performed in BO media supplemented with heparin for 2 hrs. Genes such as SLC16A1 and SLC16A7 were selected for the study. The mRNA isolated from respective samples were converted to cDNA and subjected to q-PCR analysis. The target genes were normalized to endogenous control GLUT5. The mRNA expression of genes in fresh capacitated was analyzed relative to fresh uncapacitated as control. Fold change in gene expression was calculated using ∆∆Cq method. Significant difference between groups for fold change was analyzed using t test and multiple comparison was performed by Duncan multiple range test. No significant difference (p<0.05) was observed in the mRNA expression of MCT1and MCT2 between fresh capacitated and fresh uncapacitated (control) semen samples. It was concluded that, in vitro capacitation induces no significant change in the mRNA expression of both SLC16A1 and SLC16A7 genes which encode for MCT1 and MCT2 respectively and MCTs exert minimal role in capacitation related biochemical changes in bubaline spermatozoa.
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    ThesisItemOpen Access
    EFFECT OF EXOGENOUS MELATONIN ADMINISTRATION ON EXPRESSION PATTERNS OF SELECTIVE AQUAPORIN GENES IN HEAT STRESSED BUBALINE PERIPHERAL BLOOD MONONUCLEAR CELLS
    (SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA, 2023-08) CHAITANYA .G; IQBAL HYDER (MAJOR); SRINIVASA PRASAD .CH; ASWANI KUMAR .K
    The present study was designed to evaluate the effect of exogenous melatonin administration on heat stress (HS) amelioration in buffaloes during summer season. Fifteen adult Murrah buffaloes were randomly selected from Livestock Farm Complex (LFC), NTR College of Veterinary Science, Gannavaram, Andhra Pradesh. The study was conducted during the months of May - July 2022. The animals were divided into three groups, Group-1: (GI, Control n=5), Group-2: (GII, n=5 Melatonin @18mg/50 kg bwt. Subcutaneously single administration) and Group-3: (GIII, n=5 Melatonin@18mg/50 kg b wt. subcutaneously twice at fortnight interval. The AT and RH were recorded throughout the study period and THI was calculated. Physiological parameters like RT, RR and PR were recorded daily at 1:00 PM throughout the study period. Two aliquots of blood sample (whole blood with anticoagulants and blood with clot activators for serum isolation) were collected from all the 15 animals in weekly interval for seven weeks. An aliquot of whole blood was directly used for analysis of haematological parameters (TEC, TLC, Hb and PCV) and blood glucose, while the remaining aliquot was used for separation of PBMC and subsequent RNA isolation for gene expression studies of HSP90 and selective aquaporins (AQP1, AQP8 and AQP11). Blood with clot activators was processed to separate serum, for estimation of biochemical parameters (total protein, albumin, cholesterol, ALT and AST). The THI value above 78 was observed throughout the study period indicating that the animals were under HS. The mean±SE values of RT, RR and PR didn’t vary significantly (p>0.05) between control and melatonin treated buffaloes The mean±SE values of haematological parameters didn’t vary significantly (p>0.05) between control and melatonin treated buffaloes and no significant difference (p>0.05) was observed among all the weeks of our study period. The mean±SE values of total protein and albumin did not vary significantly (p>0.05) between control and melatonin treated buffaloes and no significant (p>0.05) difference was observed among all the weeks. The blood glucose levels were significantly (p<0.05) higher in GIII compared to control in week 6 and in overall mean. The mean cholesterol concentration was significantly (p<0.05) higher in GIII compared to control during week 5, 6 and in overall mean. The AST levels were significantly (p<0.05) lowered in GIII compared to control in all the weeks and also in overall mean. The ALT levels were significantly (p<0.05) lowered in GIII during week 5, week 6 and in overall mean compared to control. The mean cortisol levels were significantly (p<0.05) lowered in GIII compared control during week 4, 5, 6 and also in overall mean. The relative mRNA expression of HSP90 and AQP8 were significantly (p<0.05) upregulated on week 2 and 3 in GII and on week 2, 3, 4 and 5 in GIII compared to control. Further, the overall expression of HSP90 was significantly (p<0.05) upregulated in melatonin treated animals compared to control. Whereas for AQP8 overall expression was significantly (p<0.05) upregulated in GIII compared GII to control. There was no significant (p>0.05) difference in the mRNA expression of AQP1 and AQP11 between control and melatonin treated groups (GII and GIII) during all the weeks of the study and also in overall. Increased expression of HSP90 in melatonin treated group compared to control in our study suggests the role of melatonin in prevention of abnormal protein aggregation and misfolding of proteins caused due to oxidative stress. The increase in expression of AQP8, a peroxyporin in melatonin treated group establishes the role of melatonin in facilitating H2O2 diffusion and efflux of other free radicals generated due to oxidative stress induced by heat stress. A THI of above 78 throughout the study period is indicative of heat stress in animals. It can be concluded from our study that Melatonin administration can substantially ameliorate the effects of heat stress in buffaloes as by altering biochemical parameters (glucose, cholesterol, AST, ALT and cortisol) and further dynamically altering expression of HSP90 and AQP8 genes.
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    ThesisItemOpen Access
    EFFECT OF EXOGENOUS MELATONIN ADMINISTRATION ON EXPRESSION PATTERN OF CERTAIN MONOCARBOXYLATE TRANSPORTER GENES IN HEAT STRESSED BUBALINE PERIPHERAL BLOOD MONONUCLEAR CELLS
    (SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA, 2023-08) SUJANA GONTLA; SRINIVASA PRASAD .CH (MAJOR); VASANTHA SESHU KUMARI .I; ASWANI KUMAR .K
    The objective of the present study was to know the effect of exogenous melatonin administration on physiological, oxidant, antioxidant and cortisol profiles along with expression pattern of MCTs and HSP70 in heat stressed Murrah buffaloes. Fifteen non pregnant adult Murrah buffaloes of 5-8 years of age were selected randomly and divided into three groups Control (n=5), T1 (n=5) melatonin@18mg/50 kg b.wt single administration and T2 (n=5) melatonin@18mg/50 kg b.wt twice at fortnight interval. The blood samples were collected by jugular vein puncture into EDTA and serum vials at weekly intervals for a period of seven weeks during study period. Oxidant, antioxidant, cortisol profiles were estimated. The physiological parameters and THI were also recorded during the period of study. THI of the present study recorded was >80 except during 5th week (THI-78). RT, RR and PR were significantly (p<0.05) lower in melatonin treated buffaloes compared to control. Significant decrease (p<0.05) was observed in RT of T2 compared to T1 while no significant (p>0.05) difference was found in RR and PR in between these groups. MDA and cortisol levels were significantly (p<0.05) decreased in T1 and T2 compared to control, while significant (p>0.05) decrease was observed T2 compared to T1. Variation in both parameters was observed in all three groups with respect to THI. The mean values of SOD, CAT and GPx were significantly higher (p<0.05) in both treatment groups compared to control, with highest values obtained (p<0.05) in T2 among all three groups. The mRNA expression of MCT 1, 2 and 8 were significantly (p<0.05) downregulated in melatonin treated groups compared to control. HSP70 expression was upregulated in treatment groups compared to control. No significant (p>0.05) difference in expression of MCT1 and HSP70 was observed between T1 and T2. The expression of MCT2 and MCT8 was significantly (p<0.05) higher in T1 than T2. In conclusion, the present study demonstrated the effective role of melatonin in ameliorating heat stress which is evident through the study on various heat stress markers like physiological parameters, estimation of oxidant and antioxidant enzyme activity, estimation of stress hormone cortisol and gene expression studies on monocarboxylate transporters MCT 1, 2 and 8 along with HSP 70. Hence, we can conclude that melatonin could be effective in ameliorating heat stress in buffaloes.