Dr. M.T. PanchalKHAWASKAR MEGHA VINOD2018-01-242018-01-242012http://krishikosh.egranth.ac.in/handle/1/5810039820This investigation was carried at the Semen Station, Anand Veterinary College, Anand (Gujarat) on a total of 60 semen ejaculates (10 ejaculates/bull/season) collected from 2 sexually mature bulls over one year period divided in to three seasons (monsoon, winter and summer). The objectives were to study the seasonal variations in the seminal characteristics including acrosomal structures and membrane integrity of sperm, and biochemical and enzymatic profile of seminal plasma, and the interrelationships among the physico-morphological, biochemical and functional attributes of semen of Surti buffalo bulls. The gross and microscopic evaluation of seminal characteristics including cold shock test, hypo-osmotic swelling (HOS) test, methylene blue reduction test and cervical mucus penetration test was carried out as per standard procedures. Initial seminal fructose was estimated by using resorcinol method. Seminal plasma biochemical and enzymatic profiles, viz., total protein, total cholesterol, AKP-ACP and GOT-GPT were determined using standard procedures and assay kits on an auto-analyzer. The data were analyzed statistically using 2 factors factorial CRD on SAS Programme to know the effect of bulls, seasons and bull x season interaction on each of the parameters studied. Finally the correlations between various physico-morphological and biochemical attributes were worked out. The overall mean ejaculate volume, consistency/density score (0-4), initial pH, mass activity (score 0-5), individual sperm motility, sperm concentration, live sperm, total abnormal sperm and sperm with intact acrosome recorded in Surti buffalo bulls’ semen were 3.16 ± 0.76 ml, 3.05 ± 0.92, 6.89 ± 0.16, 2.96 ± 0.18, 71.66 ± 12.37 per cent, 885.42 ± 242.90 millions/ml, 85.75 ± 4.65 per cent, 6.53±1.28 per cent and 90.73 ± 3.16 per cent, respectively. The bulls varied significantly only in their ejaculate volume (P<0.008), sperm concentration (P<0.002) and live sperm per cent (P<0.014). Seasonal influence was significant for ejaculate volume (P<0.002), density score (P<0.05), mass activity (P<0.002), sperm concentration (P<0.047), live sperm (P<0.001), abnormal vi sperm (P<0.003) and intact acrosome (P<0.022). The lowest values observed during winter were for ejaculate volume, sperm concentration and live sperm per cent, whereas density, mass activity and individual motility was the lowest during monsoon, and abdormal sperm per cent during summer. However, the bull x season interaction was not significant for any of these attributes. The average values for cold shock resistant (live) sperm, hypo-osmotic swelling reactive sperm, methylene blue reduction time and cervical mucus penetrance score (0-3) for Surti buffalo ejaculates were 53.45 ± 8.30 per cent, 59.28 ± 6.10 per cent, 7.11 ± 1.28 minutes and 2.20±0.74, respectively. The influence of bulls was insignificant on all these traits, however, seasonal influence was significant for cold shock resistant and hypoosmotic swelling reactive sperm (P<0.000). The lowest mean values for all four functional tests were observed during summer and the highest during monsoon, except CMPT which was highest during winter. Further, the bull x season interaction was significant only for cold shock resistant (live) sperm. The overall average seminal plasma contents of fructose, total protein, total cholesterol and enzyme AKP, ACP, GOT and GPT activity recorded were 586.61±65.77 mg/dl, 2.54 ± 0.51 g/dl, 53.71±10.05 mg/dl, 53.71±10.05 KAU/dl, 246.78 ± 27.92 KAU/dl, 57.30 ± 10.73 IU/L and 15.38 ± 5.75 IU/L, respectively. The bulls varied significantly only in their seminal plasma content of fructose (P<0.000), protein (P<0.014) and alkaline phosphatase (P<0.000). Seasonal influence was highly significant (P<0.000) for initial fructose and all four enzymes. The lowest values for fructose, protein, AKP and GPT were observed during summer and for cholesterol, ACP and GPT during monsoon. Further, the bull x season interaction was significant for seminal fructose and total proteins only. The interrelationships between various physico-morphological, functional and biochemical attributes of buffalo semen revealed that the ejaculate volume was significantly correlated with cold shock resistant sperm (0.35), MBRT (0.26), seminal fructose (0.46) and ACP (-0.35). Sperm concentration was significantly correlated with live sperm (0.30), MBRT (-0.31) and seminal AKP (-0.54). Mass activity was significantly correlated with abnormal sperm (-0.26), cold shock resistant sperm (-0.39), seminal fructose (-0.29), GOT (0.44), GPT (-0.28), AKP (-0.39) and ACP (0.26). Initial motility had significant correlation with live sperm (-0.27), HOS reactive sperm (0.25). vii Seminal pH had significant correlation with seminal cholesterol (0.26). Live sperm per cent was significantly correlated with HOS reactive sperm (-0.49), cold shock resistant sperm (-0.56) and seminal AKP (-0.41). The abnormal sperm per cent was significantly correlated with intact acrosome (0.25), cold shock resistant sperm (0.32), seminal fructose (0.33), cholesterol (-0.41), GOT (-0.32), AKP (0.27) and ACP (-0.51). The intact acrosome showed significant correlation with HOS reactive sperm (0.37) and seminal GPT (-0.33), whereas HOS reactive sperm was significantly correlated with live sperm (-0.49), cold shock resistant sperm (0.29) and seminal GOT (0.32). The cold shock resistant sperm was significantly correlated with seminal fructose (0.34), GOT (-0.36), GPT (0.29), AKP (0.59) and ACP (-0.37). The MBRT was significantly correlated only with seminal fructose (0.32). Amongst the biochemical and enzymatic attributes, significant correlations were observed for seminal fructose with seminal GOT (-0.43), GPT (0.36) and ACP (-0.38); seminal total protein with seminal GOT (-0.76); cholesterol with ACP (0.27); seminal GOT with GPT (-0.43), AKP (-0.44) and ACP (0.46); seminal GPT with AKP (0.42), and seminal AKP with ACP (-0.41). In general, the initial semen quality, in terms of physico-morphological and functional attributes, of Surti buffalo bulls under study was of good to average type and was within acceptable limit for use in artificial breeding programme. The seminal plasma profile of biochemical and enzymatic constituents studied was within the normal physiological limit for bovines and indicated optimal functioning of gonads and all the accessory sex glands. Most of the physico-morphological, functional and biochemical/ enzymatic attributes were highly significantly (P<0.001) influenced by the seasons, suggesting that Surti buffalo bulls are sensitive to seasonal variation in agro-climatic conditions of the region. Overall winter season was found to be more congenial for better quality semen production as compared to monsoon and summer season. Significant interrelationships observed between some of the physico-morphological attributes of semen and biochemical constituents of seminal plasma suggested that a good functional coordination existed between gonads and accessory sex glands in the Surti buffalo bulls towards optimizing reproductive functions, and that the assessment of motility, morphology and viability may be sufficient to predict the quality of fresh semen fairly accurately, without going into too clumsy, time consuming tests under routine laboratory procedures.enveterinary science, animal reproductionThesis