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Anand Agricultural University, Anand
Anand Agricultural University (AAU) was established in 2004 at Anand with the support of the Government of Gujarat, Act No.(Guj 5 of 2004) dated April 29, 2004. Caved out of the erstwhile Gujarat Agricultural University (GAU), the dream institution of Sardar Vallabhbhai Patel and Dr. K. M. Munshi, the AAU was set up to provide support to the farming community in three facets namely education, research and extension activities in Agriculture, Horticulture Engineering, product Processing and Home Science. At present there seven Colleges, seventeen Research Centers and six Extension Education Institute working in nine districts of Gujarat namely Ahmedabad, Anand, Dahod, Kheda, Panchmahal, Vadodara, Mahisagar, Botad and Chhotaudepur
AAU's activities have expanded to span newer commodity sectors such as soil health card, bio-diesel, medicinal plants apart from the mandatory ones like rice, maize, tobacco, vegetable crops, fruit crops, forage crops, animal breeding, nutrition and dairy products etc. the core of AAU's operating philosophy however, continues to create the partnership between the rural people and committed academic as the basic for sustainable rural development. In pursuing its various programmes AAU's overall mission is to promote sustainable growth and economic independence in rural society. AAU aims to do this through education, research and extension education. Thus, AAU works towards the empowerment of the farmers.
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ThesisItem Open Access COMPARATIVE EFFICACY OF DIFFERENT LEVELS OF ANTIOXIDANT SERICIN IN EGG YOLK TRIS EXTENDER FOR CRYOPRESERVATION OF BOVINE SEMEN(DEPARTMENT OF VETERINARY GYNAECOLOGY AND OBSTETRICS COLLEGE OF VETERINARY SCIENCE & ANIMAL HUSBANDRY ANAND AGRICULTURAL UNIVERSITY ANAND, 2019) Patel Tapasvikumar M.; Dr. A.J. DhamiThis investigation was undertaken during winter season on three mature healthy pedigreed breeding bulls each of Gir cattle and Murrah buffalo breeds, with the aim to assess effect of different concentration of antioxidant Sericin in standard Tris fructose egg yolk glycerol (TFYG) extender for improving cryopreservation of cattle and buffalo semen based on sperm quality parameters, and assay of oxidative markers in seminal plasma of freshly diluted and cryopreserved semen, and thereby to find out the optimum level of Sericin that can be recommended for cryopreservation of bovine semen. Ten ejaculates were studied from each bull at weekly interval in a split-sample technique for spermatozoa quality traits, and representative six ejaculates for oxidative markers, viz., malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity in freshly diluted and frozen-thawed seminal plasma. Only the ejaculates with >70% initial motility were used.ThesisItem Open Access MONITORING POSTPARTUM REPRODUCTIVE PERFORMANCE IN KANKREJ COWS THROUGH CLINICAL DIAGNOSIS, BLOOD PROFILE AND HORMONAL THERAPY(AAU, Anand, 2012) NAIKOO, MEHRAJUDDIN; Dhami, A. J.The present study was carried out at Livestock Research Station of the University on 42 Kankrej cows of 2nd to 4th parity. The chief objectives were: to monitor the early postpartum period (0-90 days) clinically and through plasma profile of progesterone, metabolites and macro-micro minerals at 10 days intervals; to evaluate the efficacy of a sustained release mineral supplement (Mega bolus PO) on the day of calving and five oestrus induction and synchronization protocols (Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and PGF2α) on day 90-95 postpartum towards augmenting reproductive efficiency of anestrous and subestrous cows (6 animals in each group), keeping 6 normal cyclic animals as control, and its effect on above profile till day 40 post-treatment/post- AI, and to compare plasma profiles of conceived and non-conceived cows at first Al. The time required for expulsion of fetal membranes, weight of expelled fetal membranes and the birth weight of calf (pure and crossbred) were 5.04 ± 2.0 hrs, 2.84 ± 0.76 kg and 24.29 ± 1.54 kg, respectively. The Kankrej cows showed complete uterine involution by mean interval of 36.80 ±1.21 (range 32-45) days postpartum. The interval for occurrence of first oestrus postpartum clinically and through plasma P4 profile was 105.49 ± 1.66 (range 86-106) and 56.42 ± 3.88 (range 30-80) days, respectively (P<0.05). The first service and overall conception rates obtained at spontaneous/ induced oestrus, within 150 days postpartum were 30.95 (13/42) and 40.47 (17/42) per cent. The comparative evaluation of the efficacy of five oestrus induction/ synchronization protocols tested, on 6 cows each, viz. Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and PGF2α revealed oestrus induction response of 66.66, 83.33, 50.00, 66.66 and 66.66 per cent, respectively, with behavioural signs at FTAI as confirmed by palpation per rectum. The first service conception rates obtained were 16.66, 33.33, 16.66, 50.00 and 50.00 per cent, respectively, as compared to 33.33 per cent in normal cyclic control cows. The corresponding second service conception rates were nil, 25.00, 20.00, nil, nil and nil per cent, as compared to 25.00 per cent in untreated control animals. The overall conception rates of three cycles over the 45 days period were 33.33, 50.00, 33.33, 50.00 and 50.00 per cent, respectively, as against 50.00 per cent in normal cyclic group. The results of CIDR, Cosynch and PGF2α protocols were better than the Ovsynch and normal control groups.ThesisItem Open Access EFFECT OF PROGESTERONE AND HEAT STRESS ON REPRODUCTIVE PERFORMANCE OF DAIRY COWS AND VALIDATION OF CONTINUOUS BODY TEMPERATURE MEASUREMENT IN IDENTIFYING ESTRUS(AAU, Anand, 2012) SUTHAR, VISHAL S.; Dhami, A. J.The present study with four experiments and manifold objectives were conducted on German (HF) dairy cows at or affiliated farm facilities of Clinic for Animal Reproduction, Freie Universitaet Berlin, Germany, during January 2010 to June 2012. The overall objectives of this study were to evaluate: 1) performance of temperature data logger in in vitro and in vivo conditions, 2) validity of BT to identify induced estrus, 3) effect of exogenous and endogenous P4 on BT of dairy cows and 4) and to study effect of heat stress on BT and postpartum performance of dairy cows. In first study to evaluate performance of temperature data logger (Minilog 8, Vemco Ltd., Halifax, Canada), three sub-experiments were conducted. The study began with an in vitro validation of 24 temperature loggers comparing them to a calibrated liquid-in-glass thermometer as a reference method (sub-experiment 1). The association and agreement between the 24 temperature loggers with the reference method was r = 0.996 (P < 0.001) and a negligible coefficient of variance (0.005) between the loggers. In vivo temperature loggers were tested in 11 healthy postpartum cows (sub-experiment 2) and 12 early postpartum cows with greater BT (sub-experiment 3). Temperature loggers were set to record VT and RT at 1 min intervals. To prevent rectal and vaginal straining and potential expulsion of temperature logger an epidural injection of 2.5 ml of 2% Procain was administered. Association between RT and VT was r = 0.92 (P < 0.001) in sub-experiment 2 and r = 0.94 (P < 0.001) in sub-experiment 3 with a negligible difference of -0.1 and 0.01°C, respectively. Bland-Altman plots demonstrated an agreement between RT and VT for healthy and early postpartum cows with greater BT in sub-experiment 2 and 3, respectively. Therefore, continuous VT monitoring with temperature loggers can be used as a measure for BT in dairy cows.ThesisItem Open Access INFLUENCE OF DIFFERENT CATEGORIES OF FOLLICLES ON QUANTITY AND QUALITY OF OOCYTES WITH RESPECT TO THEIR IN VITRO MATURATION IN SURTI BUFFALO(AAU, Anand, 2011) SINGH, RAHUL PRATAP; Shah, R. G.The present study was conducted on buffalo abattoir ovaries at Laboratory of Biotechnology of Veterinary College, Anand. The study was conducted over nine months from February 2010 to October 2010 on 499 ovaries. They had 1740 different size of follicles; from them 1046 different grades of oocytes were recovered for the study by slicing method. The influence of different categories of follicles on quantity and quality of buffalo oocytes, its maturation, effect of presence or absence of CL on oocytes recovery rate and its quality and in vitro fertilization of matured oocytes were studied. The total numbers of follicles recovered from 499 ovaries were 1740, of them 726 follicles were observed on the ovaries which had all the three size of follicles. The overall mean number of follicles per ovary was found to be 2.40 ± 0.07. The mean number (percentage) of small, medium and large size follicles per ovary were 3.16 ± 0.11 (49.40), 1.81 ± 0.06 (34.20), and 1.30 ± 0.05 (16.40), respectively. Oocytes collected by slicing method were classified on the basis of cumulus investment and ooplasm homogenecity, viz. grade A (>3 layers of cumulus cells), grade B (1-3 layers of cumulus cells), grade C (less compact cumulus) and grade D (nude oocytes). The mean number of oocytes per ovary of grade A (2.92 ± 0.14) and B (2.49 ±0.13) were significantly higher (P<0.05) followed by grade C (1.66 ± 0.09) than that of grade D (1.32 ±0.10) oocytes. The correlation studies between follicles size and oocytes qualities indicated that the small (<5 mm) follicles had highly significant (P<0.01) and positive correlations with grade B and C oocytes. Similarly, medium size follicles (5-8 mm) showed highly significant (P<0.01) and positive correlations with grade A oocytes. Large follicles (>8 mm) showed significant (P<0.05) and posiive correlation with grade A oocytes, whereas it had negative correlation with grade D, C, and B oocytes. The maturation rate achieved was 80.97 per cent in TCM-199 supplemented with 0.6 per cent BSA. The highest percentages of cytoplasmic maturation observed in oocytes of grade A, B, C and D were for good (50.30 per cent), fair (37.70 per cent) and poor (33.30 per cent) and poor (28.60 per cent) quality of oocytes, respectively. The significantly higher number of grade A (>3 layers of cumulus cells) and grade B (1-3 layers of cumulus cells) quality of oocytes attained good cytoplasmic maturation than grade C (less compact cumulus cells) and grade D (nude oocytes). The total maturation rate from grade A oocytes was highest (87.94 per cent) followed by oocytes of grade B (82.91 per cent) and grade C (72.22 per cent). The nuclear maturation was evaluated in 528 oocytes by Hoechst 33342 stain. The highest number of grade A oocytes (26 per cent) reached to M-II stage, followed by grade B (24.21 per cent), and grade C (14.28 per cent) and none of the oocytes from grade D reached to M-II stage, they were mainly arrested at GV and GVBD stage. The grade D oocytes did not mature and maximum per centage of degenerated oocytes (72.50 per cent) was found in this category. Of the 367 ovaries, the CL was present on 73 and absent on 294 ovaries that yielded 74 and 972 oocytes, respectively. Significantly (P<0.05) greater number of oocytes per ovary were recovered (5.06 ± 0.36) when the CL was absent compared with ovaries on which CL was present (0.38 ± 0.05). Further, significantly higher percentage (P<0.01) of recovery rate of grade A (37.76 per cent) and grade B (37.96 per cent) oocytes was obtained from the ovaries in which CL was absent than the ovaries in which CL was present (grade A: 40.54 per cent and grade B: 39.19 per cent). The effect of presence Vs absence of CL on the ovaries revealed 8.11 Vs 18.00 per cent recovery rate for grade C and 12.16 and 6.27 per cent for grade D oocytes. Oocytes of grade A (n=181) and B (n=305), which had cytoplasmic maturation, were utilized for in vitro fertilization. The overall fertilization rate observed was 20.16 per cent for grade A and B occytes. The higher fertilization rate was observed for grade A (21.00 per cent) oocytes than that of grade B (19.67 per cent). Application of in vitro embryo production technology in assisted reproduction of buffalo will not only improve productive and reproductive potential of the buffalo population but will also help to rescue the precious germ plasma going to waste by indiscriminate slaughter of this animal. This study showed that functional structure on the ovaries, i.e. corpus luteum, size of follicles etc. affect the recovery rate and quality of oocytes. The higher oocyte recovery rate and good quality oocytes were observed in absence of corpus luteum on the ovaries. It also revealed that the presence of cumulus layer around the oocytes affect the maturation rate. Further studies required for the in vitro culture system for in vitro embryo production of buffaloes.ThesisItem Open Access COMPARATIVE EVALUATION OF QUALITY AND FERTILITY OF BOVINE SEMEN USING COMPUTER ASSISTED SEMEN ANALYSER(AAU, Anand, 2011) PATEL, JIGNESHKUMAR BHEMABHAI; Dhami, A. J.This study was undertaken on 90 semen ejaculates (6/bull) of fifteen sexually mature healthy breeding bulls (5 Jafarabadi, 5 Mehsana and 5 HF x Kankrej F1), aged 3-8 years, stationed at State Frozen Semen Production and Training Institute, Patan. The study included subjective evaluation of seminal characteristics, hypo-osmotic swelling (HOS) test, post-thaw motility, and objective assessment of sperm motion characteristics of fresh and frozen-thawed semen by Computer Assisted Semen Analyser (CASA) including in vivo fertility trials of frozen-thawed semen under field conditions and establishment of interrelationships among various spermatozoal traits. The ejaculate volume (ml), mass activity (score 0-5), initial motility (%), sperm concentration (million/ml), live sperm (%), total abnormal sperm (%) and HOS reactive sperm (%) recorded in Jafarabadi buffalo semen averaged 4.81 ±0.21, 3.30 ± 0.08, 74.33 ± 0.75, 1466.80 ± 72.43, 84.93 ± 0.59, 10.67 ± 0.43 and 70.47 ± 0.54, respectively. The corresponding values for Mehsana buffalo semen were 5.01 ± 0.23, 3.23 ± 0.12, 73.83 ± 0.67, 1307.43 ± 94.27, 82.47 ± 0.67, 10.83 ± 0.38 and 71.53 ± 0.89, and for HF x K crossbred bull semen 6.68 ± 0.31, 2.83 ± 0.09, 72.00 ± 0.51, 1094.80 ± 78.85, 79.60 ± 0.65, 12.87 ± 0.45 and 61.37 ± 0.69, respectively. The values of all traits, except volume, were significantly (P<0.05) higher in buffalo bulls than the crossbreds. In Jafarabadi bulls, mass activity showed significant positive correlation with (P<0.01) sperm concentration per ml (0.865), and live sperms with abnormal sperms (-0.853). In Mehsana bulls, ejaculate volume had significant correlations with sperm concentration (-0.471) and live sperms (0.576); mass activity with initial modlity (0.554), sperm concentration (0.859) and live sperms (0.491); and sperm concentration with initial modlity (0.530) and live sperms (-0.469), whereas in HF x K bulls, mass activity showed significant correlations with initial motility (0.493) and sperm concentration (0.953); initial motility with sperm concentration (0.534), and live sperms with abnormal sperms (-0.525). The fresh semen analyzed by CASA revealed the mean motile sperm (%), progressively motile sperm (%), average path velocity (VAP μm/s), straight line velocity (VSL nm/s) and curvilinear velocity (VCL μm/s) for Jafarabadi buffalo sperms as 79.77 ± 1.62, 78.90 ± 1.22, 114.15 ± 2.28, 99.97 ± 2.09 and 181.30 ± 4.19, respectively. The corresponding values for Mehsana buffalo sperms were 61.80 ± 1.85, 61.37 ± 1.58,108.75 ± 2.59, 93.63 ± 2.14 and 176.72 ± 6.12, and for HF x K crossbreds 74.73 ± 1.71, 51.57 ± 2.61, 100.37 ± 2.61, 86.25 ± 2.15, and 158.93 ± 6.46, respectively. The values of all the traits were significantly (P<0.05) lower for crossbreds as compared to Jafarabadi and Mehsana buffalo sperms, which however differed significantly (P<0.01) for VSL. Among the other sperm velocity parameters of fresh semen, the mean values of amplitude of lateral head displacement (ALH, μm), beat cross frequency (BCF, Hz), straightness (STR, %), linearity (LIN, %), elongation (ELG, %) and sperm area (ARE, μm2) for Jafarabadi buffalo sperms were 6.57 ± 0.17, 38.51 ± 0.75, 87.63 ± 0.84, 60.73 ± 1.47, 43.07 ± 1.99 and 20.23 ± 0.58, respectively. The corresponding values for Mehsana buffalo sperms were 6.63 ± 0.27, 37.64 ± 0.64, 86.43 ± 1.04, 60.00 ± 2.58, 46.83 ± 2.41 and 22.13 ± 0.99, and for HF x K crossbreds 6.87 ± 0.31, 32.79 ± 0.81, 86.63 ± 1.13, '61.93 ± 2.24, 44.43 ± 1.51 and 22.13 ± 0.81, respectively. The mean values of none of these traits, except BCF, differed significantly between breeds. Moreover, the mean values of rapidly mofile sperms were significantly (P<0.01) higher in the semen of Mehsana and Jafarabadi buffalo bulls, whereas the values of medium motile and static sperms were significantly (P<0.01) higher in HF x K bulls. The mean post-thaw motility of Jafarabadi, Mehsana and crossbred bulls semen averaged 52.67 ± 0.79, 51.33 ± 1.01 and 50.50 ± 0.91 per cent, respectively, while HOS reactive sperms were 54.57 ± 0.61, 51.50 ± 0.83 and 46.80 ± 1.06 per cent, respectively, the values of later trait were significantly (P<0.01) higher in buffalo semen than crossbreds. For the frozen-thawed semen assessed by CASA, the viable (via-dent stained), motile and progressively motile sperms of Jafarabadi buffalo bulls averaged 87.47 ± 1.04, 51.20 ± 1.57, 33.20 ± 1.45 per cent, respectively. The corresponding values for Mehsana bulls were 84.40 ± 1.24, 52.10 ± 1.70 and 34.30 ± 1.54 per cent, and for HF x K bulls 81.37 ± 1.18, 50.80 ± 1.36 and 30.23 ± 1.16 per cent, respectively. The viable sperm differed significantly (P<0.01) only between Jafarabadi buffalo and HF x K bulls. The values of velocity parameters, viz., VAP μm/s), VSL (μm/s), VCL (μm/s), ALH (μm), BCF (Hz), straightness (%), linearity (%), elongation (%), and head area (μm2) of post-thawed sperms of Jafarabadi bulls were 71.69 ± 1.64, 55.84 ± 1.49, 126.80 ± 2.89, 6.72 ± 0.19, 34.05 ± 0.46, 74.70 ± 1.02, 44.37 ± 1.01, 50.73 ± 0.93 and 10.83 ± 0.17, respectively. The corresponding values for Mehsana bulls were 72.11 ± 1.86, 55.26 ± 1.42, 129.76 ± 3.16, 6.78 ± 0.14, 33.35 ± 0.34, 74.27 ± 0.67, 42.97 ± 0.65, 49.87 ± 0.88 and 11.20 ± 0.42, and in HF x K crossbreds 71.11 ± 1.45, 52.28 ± 1.14, 136.26 ± 2.73, 7.86 ± 0.14, 31.88 ± 0.39, 69.57 ± 0.67, 38.10 ± 0.72, 43.83 ± 0.97, and 12.03 ± 0.16, respectively. The values of VAP, VSL and VCL in frozen-thawed sperms did not differ between buffalo and crossbred bulls. The values of head area, VCL and ALH were higher in HF x K bulls than the Mehsana buffalo. The mean values of BCF, STR, LIN and ELG were statistically similar in the semen of Jafarabadi and Mehsana buffalo breeds, though differed significantly from HF x K bulls. The rapidly motile sperms in the frozen-thawed semen were higher in Jafarabadi bulls than in HF x K crossbreds. The medium and slow motile and static sperm were, however, higher in crossbreds than in buffalo bulls semen. The overall conception rates obtained through 3275, 4320 and 3632 AIs performed using frozen semen doses of respective breeds in Jafarabadi and Mehsana buffaloes and crossbred cows under field conditions were 40.37, 41.39 and 43.36 per cent, respectively, which did not differ significantly. Significant (P<0.05) or highly significant (P<0.01) interrelationships were observed between some of the CASA attributes, viz., motile, progressive motile, VAP, VSL, VCL, ALH, BCF, sperm area, rapid moving spermatozoa of Jafarabadi buffalo bulls in fresh and frozen-thawed semen (0.468 to 0.960). Similar significant interrelationships were also observed between subjective and objective (CASA) assessment traits of fresh and frozen-thawed semen of Mehsana buffalo bulls (0.464 to 0.959) and in HF x K crossbred bulls (0.465 to 0.932). These interrelationships indicated that a good functional correlation existed between the two systems of semen analysis. The CASA provided the base for quality assessment of sperm kinematics in both fresh as well as frozen-thawed semen of all three breeds and supported the visual or subjective assessment of semen quality. HOS test gave indirect indication of fertilizing potential of fresh and frozen-thawed spermatozoa.ThesisItem Open Access MONITORING POSTPARTUM REPRODUCTIVE PERFORMANCE IN GIR COWS THROUGH CLINICAL DIAGNOSIS, BLOOD PROFILE AND HORMONAL THERAPY(AAU, Anand, 2011) AMMU, RAMAKRISHNAN; Dhami, A. J.The present study was carried out at Livestock Research Station of the University on 24 Gir cows of 2nd to 4th parity. The chief objectives were: to monitor the early postpartum period (0-90 days) clinically and through plasma profile of progesterone, metabolites and macro-micro minerals at 10 days intervals; to evaluate the efficacy of estrus induction and synchronization protocols (CIDR, Ovsynch and Cosynch) on day 90 postpartum for enhancing the reproductive efficiency of anestrous and subestrous cows (6 animals in each group), keeping 6 normal cyclic animals as control, and its effect on above profile till day 40 post-AI, and to compare plasma profiles of conceived and nonconceived cows at first AI. The time required for expulsion of fetal membranes, weight of expelled fetal membranes and the birth weight of calf were 4.67 ± 0.46 hrs, 3.06 ± 0.19 kg and 19.08 ± 0.84 kg, respectively. The Gir cows showed complete uterine involution by mean interval of 36.54 ± 0.68 (range 31-42) days postpartum. The interval for occurrence of first estrus postpartum clinically and through P4 profile was 94.29 ± 2.24 (range 70-101) and 65.42 ± 5.77 (range 20-101) days, respectively (P<0.05). The first service and overall conception rates obtained at spontaneous/induced estrus were 41.66 (10/24) and 87.50 (21/24) per cent within 150 days postpartum. The comparative evaluation of the efficacy of three estrus induction/ synchronization protocols tested, on 6 cows each, viz. CIDR, Ovsynch and Cosynch revealed estrus induction response of 83.33, 83.33 and 100.00 per cent with behavioural signs, although all animals were in estrus at FTAI as confirmed by per rectal palpation. The first service conception rates obtained were 50.00, 50.00 and 33.33 per cent, respectively, as compared to 33.33 per cent in normal cyclic -control- cows. The corresponding second service conception rates were 66.66, 33.33 and 75.00 per cent, and the overall conception rates of two cycles over the 25 day period were 83.33, 66.66 and 83.33 per cent, respectively, as against 50.00 and 66.66 per cent in normal cyclic group. The results of CIDR and Cosynch protocols were better than the Ovsynch and normal control groups. The mean plasma P4 level varied significantly between different intervals in all the four groups including the overall pooled mean and also in the conceived and nonconceived groups upto day 40 post-AI. The mean P4 concentration on the day of calving was low, around 1 ng/ml, in all the groups, which further reduced slightly till day 20 postpartum and then gradually increased to reach appreciable level of 2.24 ± 1.03,2.59 ± 1.95, 2.56 ± 1.70 and 3.68 ± 1.30 ng/ml by day 30, 50, 70 and 40 postpartum in control, CIDR, Ovsynch and Cosynch groups, respectively. In control group, all the animals came into estrus at least once between day 30 and day 60 postpartum. Three animals each were inseminated on natural estrus around day 70-72 and day 80-82 postpartum. In CIDR group, three animals each remained anestrous and subestrous until day 90 postpartum. In Ovsynch group, 4 animals were in subestrous stage and two were in anestrous condition. In Cosynch group, 5 and 1 animals remained in subestrous and anestrous condition, respectively. The mean plasma progesterone concentrations of all the groups remained at the lowest or basal level on the day of AI, irrespective of whether they came to estrus naturally or after treatment. Further, the mean progesterone levels, irrespective of control or treatment groups, increased significantly by day 10 post-AI and then remained at that elevated levels till day 40 post-AI in conceived animals, whereas in non-conceived animals the levels dropped on day 20 post-AI, and thereafter showed cyclic pattern upto day 40 post- AI. The overall pooled mean values of biochemical constituents, viz. protein, cholesterol and triglyceride analyzed at 10 days interval varied in the range of 5.82 ±0.19 to 6.44 ± 0.15 g/dl, 79.66 ± 2.70 to 190.57 ± 8.84 mg/dl and 16.31 ± 0.86 to 21.53 ± 1.59 mg/dl, respectively, from calving upto 90 days postpartum. The values of cholesterol varied significantly in all the 4 groups, including pooled values, and that of triglycerides showed significant difference only in CIDR group. None of the three parameters were influenced by estrus induction/synchronization protocols, during treatment or till 40 days post-AI. The overall pooled mean values of calcium, phosphorus and magnesium studied at 10 days interval varied in the range of 8.74 ± 0.17 to 9.42 ± 0.18 mg/dl, 5.66 ± 0.18 to 7.34 ± 0.26 mg/dl and 2.87 ± 0.07 to 3.05 ± 0.08 mEq/L, respectively, from calving till 90 days postpartum. None of the values varied significantly at any of the intervals, except calcium, which showed significant difference between different intervals in control group. None of the estrus induction/synchronization protocols, viz, CIDR, Ovsynch and Cosynch used in the study influenced the plasma calcium, phosphorus and magnesium concentration. The overall pooled mean levels of micro-minerals varied non-significantly from calving until 90 days postpartum in the range of 0.78 ± 0.06 to 0.98 ± 0.06 ppm for zinc, 2.86 ± 0.19 to 3.35 ± 0.20 ppm for iron, 0.94 ± 0.06 to 1.13 ± 0.06 ppm for copper, 0.34 ± 0.02 to 0.37 ± 0.03 ppm for cobalt and 0.32 ± 0.03 to 0.38 ± 0.04 ppm for manganese. There was no effect of groups or periods on the plasma profile of these elements, except that the values of copper on day 10, 20 and 30 postpartum were significantly higher in Cosynch group as compared to other groups. None of the micro-minerals studied was influenced by the different estrus synchronization protocols used. Among the conceived and non-conceived groups of Gir cows, progesterone and total cholesterol varied significantly between 10 day intervals from calving until day 140 postpartum, including the days of treatment, AI and 10-40 post-AI in both the groups. The mean values of progesterone ranged from 0.39 ± 0.14 to 7.06 ± 1.22 ng/ml for conceived and 0.60 ± 0.13 to 5.14 ± 1.15 ng/ml for non-conceived cows, and those of cholesterol were 81.08 ± 2.44 to 209.06 ± 11.25 and 78.65 ± 4.36 to 216.58 ± 14.56 mg/dl, respectively. The mean values of magnesium differed significantly between periods in conceived group in the range of 2.73 ± 0.10 to 3.15 ± 0.12 mEq/L, whereas no such difference was noticed in non-conceived group (2.85 ± 0.08 to 3.25 mEq/L). Further, the values of P4 were significantly increased in CIDR and Cosynch groups by day 7 of treatment due to exogenous and endogenous source as compared to control or Ovsynch groups. The P4 values in conceived and non-conceived cows of all 4 groups were lowest on the day of AI, with peaks on day 10 post-AI, which were then maintained almost at the same levels in the conceived groups till day 40 post-AI, while in nonconceived groups they dropped significantly on day 20 post-AI and then showed cyclic pattern till day 40 post-AI. However, none of the other parameters studied, viz., plasma proteins, triglycerides, calcium, phosphorus and trace minerals, varied significantly between periods within the conceived/non-conceived group or between the two groups at any of the intervals postpartum. The study, in general, revealed that CIDR and Cosynch protocols were better in estrus expression and fertility, and thus, reduce the maintenance cost of dry animals, which in fact is of economic importance to the farmers. The plasma progesterone profile studied at 10 days interval postpartum helped in detecting silently cycling animals. The biochemical investigations helped to conclude that the selected animals in all hormone protocols and control group were healthy and were maintained under optimum nutritional regime, and this was probably the reason, why their profile was not influenced by the various estrus induction and synchronization protocols used.ThesisItem Open Access STUDIES ON RETENTION OF FETAL MEMBRANES WITH REFERENCE TO BLOOD PROFILE, CLINICAL FACTORS AND POSTPARTUM FERTILITY IN SURTI BUFFALOES(AAU, Anand, 2011) THAVANI, KAMLESH LOKCHAND; PATEL, J. A.The present investigation was carried out to know the effect of clinical factors on the incidence of Retention of Fetal Membranes (RFM) and the effect of RFM on blood profile and postpartum fertility of the Surti buffaloes. Fifty Surti buffaloes with RFM from the farmers of Anand district, Gujarat, were selected to evaluate the clinical factors affecting RFM. For blood profile, 37 animals of farmers as well as of University farm, Anand were selected and divided into three groups, viz., Group I- Retention of fetal membranes for more than 12 hours after parturition (RFM/AP, n=12); Group II- Retention of fetal membranes for more than 12 hours after abortion (RFM/AA, n=10) and Group III- Expulsion of fetal membranes within 12 hours of parturition (Control, n=15). The incidence of RFM as per data collected from Amul Dairy, Anand for the period of March 2010 to February 2011 was 46.37 per cent of the total reproductive disorders (59267) and as 4.81 per cent of the total cHnical cases (571080) recorded. Among clinical factors season, nutrition, birth weight of calf and parity had an effect on the incidence of RFM. The highest incidence of RFM was noticed during monsoon (45.52 %) followed by winter (32.66 %) and summer (21.82 %). Incidence of RFM decreased with the increase in the quantity of concentrates feeding. With the increase in the birth weight of calf, the per cent occurrence of RFM also increased. Incidence of RFM was highest in the buffaloes during third lactation followed by second and fourth lactations. No effect of previous parturient abnormalities and suckling or weaning was observed on the incidence of RFM. Pooled serum total protein levels on day 30 and 45 were significantly (P<0.05) higher than those observed on day 0. The pooled mean serum total protein level observed in control group was significantly (P<0.05) higher than that observed in RFM/AA group. The pooled mean serum cholesterol level on day 45 was significantly (P<0.05) higher than the levels observed on day 30 and 0, the latter two also differed significantly (P<0.05). The pooled mean blood glucose level on day 0 was significantly (P<0.05) higher than that observed on day 45. The pooled mean blood glucose level observed in control group was significantly (P<0.05) higher than that observed in RFM/AA group. The pooled mean serum aspartate aminotransferase activity on day 0 was significantly (P<0.05) lower than those observed on day 30 and 45. The pooled mean serum alanine aminotransferase activity observed in control group was significantly (P<0.05) higher than that observed in RFM/AP and RFM/AA groups. The values varied significantly (P<0.05) between periods, irrespective of groups, for alanine aminotransferase. The pooled mean serum calcium level observed in control group was significantly (P<0.05) higher than that obtained in RFM/AP and RFM/AA groups. The pooled mean serum calcium level at day 45 was significantly (P<0.05) higher than that observed on day 0, but did not differ significantly from that recorded on day 30. The pooled serum inorganic phosphorus level on day 45 was significantly (P<0.05) higher than that observed on day 30 and 0, and the latter two also differed significantly (P<0.05). However, the group and period effect was found non-significant for serum magnesium and Ca : P ratio. The estimation of serum micro-minerals revealed that the pooled mean serum zinc level on day 45 was significantly (P<0.05) higher than that observed on day 30 and 0. The pooled mean serum manganese level on day 0 was significantly (P<0.05) lower than that observed on day 30 and 45. However, no significant difference was observed in mean serum iron and copper contents among groups and periods studied. Endocrinological estimation revealed that no significant difference was observed for mean serum progesterone concentration on day 0, 30 and 45 among the groups studied. The number of services per conception and fertile estrus interval (days) in RFM/AA group were significantly (P<0.05) higher than the control group, but did not differ stastically from those observed in RFM/AP group. In general, season, nutrition, birth weight of calf and parity influenced the incidence of RFM. Moreover, estimation of serum total protein, serum total cholesterol, blood glucose, serum calcium and serum zinc helps in predicting the RFM in buffaloes.ThesisItem Open Access COMPARATIVE EFFICACY OF EGG YOLK AND SOYA BASED EXTENDERS FOR REFRIGERATION (5 °C) AND CRYOPRESERVATION (-196 °C) OF BUFFALO SEMEN(AAU, Anand, 2014) Dineshkumar Chaudhary V.; Dr A.J. DhamiThe present investigation was undertaken during the favourable breeding season (November-February) of the year 2013-2014 on six mature Surti buffalo bulls at Central Sperm Station of Department of Gynaecology and Obstetrics of the Veterinary College, AAU, Anand. The study covered evaluation of seminal characteristics in neat semen and then comparative efficacy of egg yolk based standard TFYG (Tris-citric acid-fructoseegg yolk-glycerol) extender and soybean based commercially available extenders (Bioxcell® and Optixcell®, IMV, France) using split-ejaculate technique through various morphological and functional attributes of spermatozoa extended/preserved/ processed in these extenders for refrigeration preservation (at 5°C up to 72 hrs) and cryopreservation (-196°C), including interrelationships of quality sperm parameters of fresh, refrigerated and cryopreserved semen. Immediately after collection, the ejaculates (8 per bull) were evaluated for routine physico-morphological attributes, including motility, viability, morphology (eosinnigrosin), acrosomal integrity (Giemsa stain) and plasma membrane integrity (HOST 150 mOsm/L; an in vitro fertility test) through standard procedures and using phase contrast microscope. The ejaculates were divided in to three equal aliquots, and extended at the concentration of 100 ×106 spermatozoa ml-1 at 34°C with 3 different extenders. Small portions of the extended semen samples (2 ml from each aliquot) were transferred to a refrigerator for 5°C preservation and evaluated at 24 hrs interval up to 72 hrs for above quality parameters. The remaining portions of extended semen samples were used for filling the French mini straws on IS4 system (IMV, France). After gradual cooling over 60-90 minutes and equilibration for 4 hrs in cold handling cabinet, the straws were frozen in liquid nitrogen vapour using a programmable bio-freezer (Digitcool 5300 CE ZH 350, IMV, France). The straws of all three extenders were evaluated at pre-freezing (after equilibration) and after 24 hrs of freezing (post-freeze stage) for the above quality parameters. Post-thaw incubation test (37 ºC) was also performed to evaluate sperm survival at 30 and 60 min of incubation. The mean values of ejaculate volume, density (1-4 score), sperm concentration, mass activity (0-5 score), individual sperm motility, live sperm, intact acrosome and HOS reactive sperms observed in fresh semen were 3.43±0.10 ml, 2.14±0.11, 744.90± 36.38 million/ml, 3.45±0.07, 78.54±0.51 per cent, 90.48±0.33 per cent, 94.40±0.20 per cent and 79.35±0.42 per cent, respectively. The variation between bulls was significant (P<0.05) for most of these traits. The overall segment-wise per cent abnormalities of sperm head, midpiece and tail region recorded in fresh semen were 1.83±0.09 0.88±0.06 and 3.44±0.08, respectively, with the total of 6.15±0.15 per cent. The mean percentages spermatozoa having swollen, ruffled, detached and denuded acrosome in fresh semen were found to be 2.29±0.08, 1.60±0.10, 1.10±0.09 and 0.60±0.08, respectively. The overall pooled mean percentages of progressively motile spermatozoa (irrespective of extenders) observed on dilution (0-hr), and after 24, 48, 72 hrs of refrigeration preservation of semen at 5°C were 78.54±0.30, 68.30±0.30, 59.38±0.35 and 51.63±0.47. The corresponding values for live sperm per cent were 90.48±0.19, 76.89± 0.51, 69.51±0.35 and 61.86±0.45; those of morphologically normal sperm per cent 93.85±0.08, 90.70±0.11, 89.08±0.10 and 87.66±0.10; intact acrosome per cent 94.40± 0.12, 88.22±0.13, 84.60±0.14 and 81.70±0.17, and HOS reactive sperm (intact plasma membrane) per cent 79.35±0.24, 65.99±0.31, 57.69±0.36 and 50.70±0.43, respectively, all of which differed highly significantly (P<0.01) between storage intervals. The mean percentages of progressively motile spermatozoa observed in Optixcell, TFYG and Bioxcell extenders at 24, 48, 72 hrs of refrigeration storage were 70.31±0.46, 68.33±0.45, 66.25±0.46; 61.46±0.51, 59.79±0.58, 56.88±0.55, and 53.85± 0.75, 52.50±0.68, 48.54±0.84, respectively. The corresponding values for live sperm per cent in Optixcell, TFYG and Bioxcell extenders at three intervals were 78.35±1.36, 77.35±0.46, 74.96±0.47; 71.96±0.48, 69.90±0.54, 66.67±0.54, and 64.38±0.70, 62.56± 0.65, 58.65±0.78, and those of morphologically normal sperms were 91.37±0.16, 90.65±0.15, 89.79±0.14; 90.08±0.15, 88.20±0.11, 88.25±0.12, and 88.54±0.12, 87.35± 0.14, 87.08± 0.15, respectively. The mean percentages of spermatozoa with intact acrosomes observed in Optixcell, TFYG and Bioxcell extenders at 24, 48, 72 hrs of refrigeration storage were 89.27±0.20, 88.15±0.18, 87.25±0.21; 85.58±0.24, 84.63± 0.18, 83.60±0.24, and 82.83± 0.27, 81.67±0.23, 80.60±0.30, respectively. The corresponding values for sperms with intact plasma membrane (HOS reactive sperms) in Optixcell, TFYG and Bioxcell extenders at three intervals were 68.10±0.46, 66.04±0.50, 63.81±0.45; 60.00±0.55, 57.92±0.55, 55.15±0.56, and 53.40±0.68, 51.35±0.60, 47.35±0.68, respectively. The results were significantly (P<0.05) superior with Optixcell followed by TFYG extender than with the Bioxcell. There were gradual and significant (P<0.05) decline in progressively motile, viable, morphologically normal, acrosomal intact and plasma membrane intact sperm per cent in all the three extenders with each increase in 24 hourly storage interval at 5°C. The overall pooled mean percentages of progressively motile spermatozoa observed (irrespective of extenders) at initial, pre-freeze and post-thaw stage of buffalo semen were 78.54±0.30, 69.48±0.26, and 47.33±0.52. The corresponding values for live sperm per cent were 90.48±0.19, 79.39±0.28 and 56.90±0.52; those of morphologically normal sperm per cent 93.85±0.08, 92.14±0.10 and 87.88±0.12; intact acrosome per cent 94.40±0.12, 89.55±0.14 and 77.08±0.17, and HOS reactive sperm (intact plasma membrane) per cent 79.35±0.24, 67.94±0.26 and 45.05±0.52, respectively, all of which differed significantly (P<0.01) between freezing stages. The overall mean post-thaw incubation (37°C) survival of spermatozoa immediately after thawing (0-min), and after 30- and 60-min of post-thaw incubation was 47.33±0.85, 41.67±0.85 and 34.62±0.82 per cent (P<0.01), respectively. The mean percentages of progressively motile spermatozoa observed in Optixcell, TFYG and Bioxcell extenders at pre-freeze and post-thaw stages were 70.94± 0.38, 69.48±0.37, 68.02±0.49, and 49.90±0.90, 47.71±0.79, 44.38±0.85, respectively. The corresponding values for live sperm per cent in Optixcell, TFYG and Bioxcell extenders at pre- and post-freeze stage were 81.58±0.38, 79.21±0.39, 77.38±0.48, and 59.67±0.91, 57.19±0.79, 53.85±0.84; and those of morphologically normal sperms 92.92±0.15, 92.10±0.14, 91.40±0.16, and 88.73±0.18, 87.67±0.17, 87.25±0.21, respectively. The mean percentages of spermatozoa with intact acrosomes observed in Optixcell, TFYG and Bioxcell extenders at pre-freeze and post-thaw stage were 90.52± 0.21, 89.54±0.18, 88.58±0.22, and 78.50±0.25, 76.83±0.23, 75.90±0.27, respectively. The corresponding values for sperms with intact plasma membrane (HOS reactive sperms) in Optixcell, TFYG and Bioxcell extenders at pre-freeze and post-thaw stage were 70.23±0.37, 67.96±0.32, 65.65±0.42, and 47.81±0.90, 45.02±0.84 and 42.31±0.82, respectively. The results were significantly (P<0.05) superior in Optixcell followed by TFYG extender as compared to the Bioxcell extender. The mean percentages of progressively motile spermatozoa observed immediate after thawing in TFYG, Bioxcell and Optixcell extenders were 47.71±0.79, 44.38±0.85 and 49.90±0.90, respectively. The corresponding motility values after 30-min of postthaw incubation were 41.98±0.80, 38.44±0.82 and 44.58±0.93 per cent, and those of 60- min of incubation were 35.52±0.79, 31.15±0.85 and 37.19±0.81 per cent, respectively. The results showed that the commercial soya based Optixcell as well as standard egg yolk based TFYG extenders could sustain better and acceptable level of sperm survival at least for 1-hr after thawing. The overall mean percentages of sperms with abnormal head, mid-piece and tail and those with different forms of acrosomal defects like swollen, ruffled, detached and denuded acrosome also followed the pattern of morphologically normal sperm and intact acrosomes during different intervals of refrigeration as well as cryopreservation of extended buffalo semen in different extenders. The values of all these traits differed significantly (P < 0.05) between storage intervals, and increased with increase in storage duration at 5°C or in post-thawed over pre-freeze stage. The sperm protective ability of Optixcell extender was superior followed by TFYG and the least of Bioxcell. The pre-freeze and post-thaw sperm motility, viability, morphology, acrosomal integrity and plasma membrane integrity recorded in all three extenders followed the same trend of declining in each extender at post-thaw over pre-freeze stage. Optixcell was significantly superior, and at par with TFYG, than the Bioxcell in maintaining greater motility, viability, morphology, acrosomal / plasma membrane integrity of buffalo spermatozoa during cryopreservation process as well as in terms of keeping quality at refrigeration (5°C) preservation also. There were significant (P<0.01) interrelationships between sperm motility, viability, normal morphology, intact acrosome and plasma membrane integrity in fresh, refrigerated and cryopreserved buffalo semen, which proved that initial motility and membrane integrity can be used as predicative measures in routine semen evaluation.ThesisItem Open Access “COMPARATIVE EVALUATION OF QUALITY AND FERTILITY OF BOVINE SEMEN USING COMPUTER ASSISTED SEMEN ANALYSER”(Anand Agricultural University, Anand, 2011) PATEL JIGNESHKUMAR BHEMABHAI; Dr. A.J. DhamiThis study was undertaken on 90 semen ejaculates (6/bull) of fifteen sexually mature healthy breeding bulls (5 Jafarabadi, 5 Mehsana and 5 HF x Kankrej F1), aged 3-8 years, stationed at State Frozen Semen Production and Training Institute, Patan. The study included subjective evaluation of seminal characteristics, hypo-osmotic swelling (HOS) test, post-thaw motility, and objective assessment of sperm motion characteristics of fresh and frozen-thawed semen by Computer Assisted Semen Analyser (CASA) including in vivo fertility trials of frozen-thawed semen under field conditions and establishment of interrelationships among various spermatozoal traits