Browsing by Author "ANJALI AGGARWAL"
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ThesisItem Open Access ANALYSIS OF SEMEN QUALITY AND FERTILITY BIOMARKERS IN CROSSBRED MALE GOATS SUPPLEMENTED WITH ZINC DURING DIFFERENT SEASONS(ICAR-NDRI, KARNAL, 2022) RESHMA DEBBARMA; ANJALI AGGARWALThe health of livestock is significantly impacted by heat stress, metabolic dysfunction, oxidative stress, and immunological suppression as a result of more frequent extreme weather events, including higher temperatures and extremely low temperatures. The present study was experimentally investigated (i) to study the effect of zinc supplementation on the semen parameters and fertility biomarkers in seminal plasma of crossbred male goats during different seasons (ii) to study the effect of zinc supplementation on haematological parameters, hormones and enzymes in blood of crossbred male goats during different seasons. To investigate, eighteen Alpine x beetle buck goats age (2-4 years) were selected and classified into three groups (n=6) as control, zinc supplemented group T1 @40ppm/Kg BW DMI and T2 @60ppm/Kg BW DMI on the basis of their average body weight. All bucks were housed in individual pens and fed in accordance with ICAR standards uniformly. In each season, semen samples and blood were collected and physiological parameters were observed fortnightly from mid December to June. The influence of season (winter, spring and summer) and effect of zinc supplementation on animal’s physiology, semen and blood parameters were studied. Season showed significant (p˂0.05) variations in semen parameters. Semen quality in the zinc supplemented group was improved as evidenced by a statistically significant (p˂0.05) increase in semen volume, progressive motility, individual motility, and acrosomal integrity. The activities of seminal plasma glutathione peroxidase, superoxide dismutase, catalase, and MDA were all significantly (p˂0.05) varied during different seasons and were improved by zinc supplementation as well as MDA level reduced. There was a significant (p˂0.05) seasonal variation in heart rate, respiration rate, rectal temperature, TEC, TLC, PCV, haemoglobin, ALT, AST and lactoferrin. The levels of blood plasma cortisol, thyroxine, zinc, testosterone, ALT and AST were significantly different (p˂ 0.05) in different seasons. Therefore this study concludes that seasonal variation affects the semen quality. Zinc supplementation improved the semen quality by reducing oxidative stress in crossbred male goats. Seminal attributes, testosterone hormone, enzymes such as glutathione peroxidise, catalase, malondialdehyde and seminal plasma protein lactoferrin can serve as potential fertility biomarkers.ThesisItem Open Access ASSESSMENT OF MILK METABOLOMICS AND PHYSIOBIOCHEMICAL PROFILES IN ZINC SUPPLEMENTED DAIRY GOATS DURING WINTER AND SUMMER SEASONS(ICAR-NDRI, KARNAL, 2022) POOJA ANAND; ANJALI AGGARWALPresent study was carried out on crossbred lactating goat (Capra hircus) to study the effect of zinc supplementation on milk metabolites and physio-biochemical changes during winter and summer seasons. Eighteen crossbred Alpine × Beetal (AB) goats were selected and divided into three groups, 6 animals each as Control (C: no zinc supplementation), Treatment 1 (T1: 30ppm of zinc supplementation) and Treatment 2 (T2: 50ppm of zinc supplementation) on DMI basis during winter and summer seasons. Milk and blood samples were collected at fortnight interval. The physiological, hematological, bio-chemical and hormonal parameters as well as milk yield, milk composition and milk minerals content in goat were investigated during both the seasons. The present study intended to investigate the changes in milk metabolites if any in zinc supplemented dairy goats. One-way ANOVA analysis revealed no significant difference between the metabolites in milk samples of control, T1 and T2 groups. However, PLS-DA analysis revealed twenty-five (25) metabolites found in control, T1 and T2 groups of milk samples were having VIP score ≥ 1. Based on -log10(p) and impact values, pathways such as alanine, aspartate and glutamate metabolism, biosynthesis of unsaturated fatty acid, TCA cycle, linoleic acid metabolism, glyoxylate and dicarboxylate metabolism, glutathione metabolism, and glycine, serine and threonine metabolism were found to be differentially regulated in control, T1 and T2 groups and these were involved in carbohydrate, lipid, amino acid and nucleotide metabolism. No significant effect of zinc supplementation was observed on rectal temperature, respiration rate, pulse rate and skin temperature in control, T1 and T2 groups but significantly (p<0.05) higher values were observed during summer as compared to winter. No significant effect of zinc supplementation was observed on hematological parameters (Hb, PVC and TEC), milk yield and milk composition (fat, SNF, protein and lactose) but their values were significantly (p<0.05) higher in control, T1 and T2 groups during winter as compared to summer. No significant effect of zinc supplementation and seasonal variation was observed on milk mineral content of sodium, potassium and zinc but significantly (p<0.05) higher values of milk calcium were observed in control, T1 and T2 groups during winter as compared to summer. Plasma cortisol was found significantly (p<0.05) lower in T2 group during summer season, whereas plasma thyroxine was found significantly (p<0.05) higher in T2 group during winter season. A significantly (p<0.05) higher level of plasma insulin was observed in control and treatment (T1 and T2) groups of AB goats during summer season, whereas no effect of zinc supplementation and seasonal variation was observed on plasma prolactin levels. No significant change in plasma glucose level was observed in control and supplemented groups (T1 and T2) but during winter season, significantly (p<0.05) higher plasma glucose levels (mg/dl) was observed. A significant (p<0.05) increase in plasma zinc levels in T2 group, during the IV and V fortnight intervals of experimental period during summer season was observed. A significantly (p<0.05) higher values of plasma copper level (ppm) in control group as compared to T2 group, whereas no significant variation between the supplemented groups (T1 and T2) was found during summer season. Based on present study, it can be concluded that supplementation of 50ppm of zinc was better in ameliorating effect of heat stress in goats.ThesisItem Open Access DEVELOPMENT OF ELISA FOR ESTIMATION OF BOVINE ANTI-MULLERIAN HORMONE USING EPITOPE SPECIFIC ANTIBODY(ICAR-NDRI, KARNAL, 2022) PRASANNA PAL; ANJALI AGGARWALAnti-Mullerian Hormone (AMH) is a dimeric glycoprotein and a member of the transforming growth factor β (TGF-β) family of growth and differentiation factors. AMH secretion is not dependent on other hormones and it can be used potentially to predict fertility in cattle. So, assessment of AMH levels in animals can be beneficial for the selection of animals. Unfortunately, generating polyclonal and monoclonal antibodies used in immunological assays have several disadvantages. The problem can be solved through selection of epitopes from the AMH protein and generating antibody against those. Therefore, we aimed to generate antibodies against selected epitope specific peptide sequence of bovine AMH; to develop and validate a suitable bovine AMH assay using raised antibodies and to estimate the basal level of AMH in indigenous cows in different age groups. For this purpose, we predicted three linear B cell epitope and synthesized three peptides. The peptides were conjugated to Keyhole limpet hemocyanin (KLH) and emulsified with FCA (Freund’s Complete Adjuvant) or FIA (Freund’s Incomplete Adjuvant). Nine female rabbits of three months of age were selected and 250 μg of peptides were administered in the rabbits (n=3) through subcutaneous route with a volume of 0.5 ml (day 0, 14, 28, 42). After final dose, we collected sera, determined antibody titer, purified it and conjugated with HRP. A good titer was observed for AMH 1 and AMH 3 peptides but not in AMH 2. The purified Ig G antibody concentration in serum was found to be 8.005 mg/ml and 10.7725 mg/ml for peptide 1 and 3 group respectively. The antibodies were used for development of direct, competitive and sandwich ELISA. Among three ELISA, sandwich was found to be the best and selected for further study. The assay was highly specific as it did not bind with AMH-related members and with non-related members. The sensitivity of direct and competitive ELISA was 50 pg/ml and for the sandwich ELISA was 5.0 pg/ml. The intra-assay CV was <6% and the inter-assay CV was <9%. The average recovery percentages were found to be within 88-100%. LLOQ was found to be 5 pg/ml and ULOQ at 50 μg/ml (CV< 20%). Blood samples (5 ml) were collected from Sahiwal and Tharparkar cattle of 0-10 years of age for standardizing the level of AMH. The concentration of AMH changes with progression of age in both the breeds (p<0.001) but no difference was observed between the breeds (p>0.05). The minimum AMH level was found in 0 to six months of age for both the breeds. Afterwards it increased significantly (p<0.05) and reaches a peak value between 1.5-2.5 years. After attaining the peak value, the level of AMH remained nearly same up to about eight years of age (p>0.05) with slight fluctuations. Thereafter it decreased progressively and reached lowest value at ten years of age. In conclusion, we developed a new highly sensitive ELISA against bAMH using epitope specific antibodies and performed age specific characterization of bAMH in Sahiwal and Tharparkar breeds for the first time.