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ThesisItem Open Access EVALUATION OF ANTI-DIABETIC EFFECT OF CAMEL MILK PROTEIN HYDROLYSATES(ICAR-NDRI, KARNAL, 2019) KHAN, SUHAIL HAKEEM; Kapila, SumanCamel milk has been reported to have potential benefits in many health issues and diseases with antibacterial, anticarcinogenic, antioxidant, antihpertensive and antidiabetic properties. The exact antidiabetic component(s) present in camel milk is still unknown. Therefore, the present study has been designed to evaluate the antidiabetic potential of camel milk protein hydrolysates. Camel milk protein hydrolysates were prepared using different digestive enzymes and among them the pepsin hydrolysates of whey proteins stimulated the pancreatic beta cell proliferation and insulin secretion by RIM-5F cell lines significantly All the hydrolysates of camel casein and whey proteins exhibited DPP-IV inhibitory activity. The antidiabetic effect of pepsin hydrolysates of whey proteins (WPH), whey proteins (WP), casein proteins (CP), camel milk (CM) was carried out in the diabetic rat model. Hyperglycemic conditions in experimental rats were created by intraperitoneal injection of streptozotocin. The administration of camel milk to diabetic rats for two months resulted in significant enhancement in body weight. There was significant decrease in the levels of blood glucose, HbA1c and increase in the insulin levels in the diabetic rats maintained on whey protein hydrolysates based diet. Camel milk and its protein and protein hydrolysates were effective in decreasing the levels of plasma TC, TGs, VLDL-C, LDL-C, atherogenic index and increasing the levels of HDL-C in the diabetic rats. The hepatic cholesterol and TGs were reduced significantly on feeding CM, WP and WPH whereas only CM and WPH helped to increase cholesterol and TGs excretion in feces significantly. Moreover, WPH also helped to improve liver antioxidative enzymes (Catalase and SOD). The kidney function markers were improved in the diabetic rats by the administration of WPH as depicted by decreased levels of urea, uric acid and creatinine in plasma. CP, WP and WPH also decreased the urea level in urine & WP and WPH increased the levels of uric acid in urine but excretion of creatinine significantly increased with the treatments of CM and WPH. Liver enzyme AST was significantly reduced in groups treated with CM, CP, WP and WPH and treatment of WP and WPH decreased ALP enzymes significantly. Expression of gluconeogenesis enzyme gene (pepck) and bile acid synthesizing gene (CYP7A1), HMGCoA and g6pase in liver were significantly downregulated in CM, CP, WP & WPH. Thus all the in vitro and in vivo results indicated that among different interventions camel WPH has the maximum antidiabetic potential.ThesisItem Open Access ASSOCIATION OF SELECTIVE GENES FROM EARLY LACTATION STRESS SPECIFIC ADIPOSE TISSUE WITH POST-PARTUM ANESTRUS IN BUFFALOES(ICAR-NDRI, KARNAL, 2021) SURYA KANT VERMA; SUNEEL KUMAR ONTERUGenetic markers associated with postpartum anestrus interval (PPAI) of buffaloes need an extensive exploration for implementing them in marker assisted selection programs. Therefore, the present study identified 29 SNPs in 11 candidate genes using pooled DNA of Murrah buffaloes with extreme positive (case) and negative (control) PPAI residuals resulted by adjusting the recorded PPAI with 38 nongenetic variables collected in the field conditions. Considering the conservative bioinformatics based functional criteria, only 7 SNPs located in the AKR1C3, PLG, MYF5, SEC14L2, TPM1, LAMA2 and TRHDE genes were selected for association analysis. During the discovery phase, the SNPs in LAMA2 (c.+549967C>A) and TRHDE (c.+445G>C) genes were significantly associated (P=0.02267 and P=0.0021, respectively) with PPAI in a population sample of 50 extreme cases and 50 controls. The significant association of the SNP in LAMA2 (c.+549967C>A) gene was further confirmed (P=0.0401) in a validation population of 415 buffaloes, in which the SNP explained 1.36% of genetic variance and the CC genotypes had 12.71 ± 3.21 PPAI days greater than that of AA and CA genotypes. Computational analysis showed the RNA transcribed from C allele of the LAMA2 (c.+549967C>A) gene might have higher stability than that of A allele. This prediction was confirmed by higher LAMA2 gene (P = 0.01798) expression (> 8 fold) and fat percentage (P < 0.05) in the adipose tissue of CC genotypes (48.78 ± 1.87%) than AA genotypes (33.59 ± 4.5%). Similarly, significantly (P<0.05) higher upregulation of the CEBPβ, PPARγ, FASN, CEBPα, PNPLA2 and the downregulation of BMP2 and PTCH1 genes in LAMA2 (c.+549967C>A) CC genotypes than AA genotypes, indicating the role of PPARγ mediated pathway for both the adipogenesis and lipolysis in CC genotypes. Overall, we found an association of the LAMA2 (c.+549967C>A) and TRHDE (c.+445G>C) SNPs with PPAI in Murrah buffaloes, which needs future validation in large populations.ThesisItem Open Access METABOLOMIC ANALYSIS OF BUFFALO FAECES TO IDENTIFY ESTRUS AND EARLY PREGNANCY BIOMARKERS(ICAR-NDRI, KARNAL, 2019) SHARMA, RITU; SINGH, DHEERFaeces play significant roles in chemical communication in many animal species. Faeces is believed as good non-invasive medium for various diagnostic purposes. In the present study, a comprehensive metabolomics and metagenomic analysis of buffalo faeces during the estrous cycle and early pregnancy was studied. Results showed total three bacterial phyla with six different bacterial order having twenty-four different genera were identified during the estrous cycle. In addition, the Clostridiales were found to be most abundant at estrus while the Bacteroidales were encountered exclusively at estrus. Pathway enrichment analyses showed that steroid biosynthesis and steroidogenesis were most enriched metabolic pathways at estrus. Testosterone propionate, 17-alpha-methyltestosterone, hydrocortisone acetate, cortisone, corticosteroid ester, testosterone cypinate, pregneninolone, aldosterone, carticosteron, glucocortocoids, testosterone enanthate, cholesterol benzoate and cholesterone were most predominant metabolites during estrous cycle with higher abundance intensity at estrus. Based on the above interpretation, it has been postulated that the cycling dairy animals have a close relationship between steroid metabolites and members of order Clostridiales and Bacteroidales. Unfortunately, we did not further validate the individual steroid(s) and/or metabolites in the faeces. Thus, it is difficult to establish a direct link between faecal steroid hormone metabolites and faecal bacteria during the estrous cycle. In addition, 9 metabolites like Pseudoephedrine, 2,5,8,11,14,17- Hexaoxaoctadecane, 1,3-Dioxolane, 2-heptyl-, Silane, (bromomethyl)trimethyl-, Benzaldehyde, 3-(4-methoxyphenoxy)-, Benzene, 1-bromo-2-fluoro-, Juglone, 4- t-Butyl-O-xylene, and 1-Naphthalenamine, N-phenyl- were identified exclusively at estrus only. However, the direct role of these metabolites in estrus in not yet known. Total five bacterial phyla with nine different bacterial order having thirtyeight different genera were identified during early pregnancy in buffalos. The phylum Firmicutes were found to be common and most abundant in both pregnant and non-pregnant animals. The proportion of order Clostridiales was found to be higher in pregnant animals than the non-pregnant, while the order Acholeplasmatales, Erysipelotrichales and Rhizobiales was exclusively present in pregnant animal. Interestingly, the free fatty acids were found to be high in pregnant animal in compare to non-pregnant. Metabolomic data also showed that the fatty acid biosynthesis and glycerosphingolipid biosynthesis pathways were highly enriched in pregnant animals. Hexadecanoic acid, Dodecanoic acid, Tetradecanoic acid, undecanoic acid, nonadecanoic acid, octadecanoic acid, pentadecanoic acid and tridecanoic acid were highly abundant in pregnant buffaloes. These differentially abundant free fatty acids could be the potential biomarker for early pregnancy. In conclusion, the results of present study demonstrated that shift in faecal metabolome and bacterial profile during the estrous cycle and early pregnancy showed a strong relationship amongst the gut microbiome, faecal metabolome and animal reproductive physiology.
