Loading...
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.
Browse
11 results
Search Results
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 STUDIES ON PHYSICO-BIOCHEMICAL CHARACTERISTICS OF ESTRUAL CERVICO-VAGINAL MUCUS WITH REFERENCE TO BODY CONDITION SCORE AND FERTILITY IN RURAL CROSSBRED COWS(AAU, Anand, 2010) GAVIT, SANJAYKUMAR PUNDALIKBHAI; Panchal, M. T.The present study was undertaken at the Department of A.R., Gynaecology and Obstetrics during the period from November 2009 to April 2010, on the rural crossbred cows (n=100) selected at random from the Ambulatory Clinic Centers, viz., Chikhodra (n=70), Bedva (n=05) and Sarsa (n=25) of Anand district. Incidence of fertile and infertile heats in cows A total of 100 breedable crossbred cows, being presented for AI, were screened per rectum together with collection of history. The per cent incidence of fertile and infertile heats in cows being attended at Chikhodra, Bedva and Sarsa were found to be 58.57 and 41.43; 40.00 and 60.00, and 56.00 and 44.00, respectively, with the overall per cent incidence of fertile and infertile heats in the different villages to be 57.00 and 43.00, respectively. Physical characteristics of CVM in conceived and non-conceived cows The per cent clear, turbid and dirty colours of CVM were obtained to be 92.98, 1.76 and 5.26; and 79.07, 2.33 and 18.60, respectively, in conceived and non-conceived cows, with the difference being non-significant. The overall per cent values for clear, turbid and dirty CVM were obtained to be 87.00, 2.00 and 11.00, respectively. The per cent thin and thick consistencies of CVM were found to be 89.47 and 10.53; and 58.14 and 41.86, respectively, in conceived and non-conceived cows, with the difference found to be significant (P<0.01). The overall per cent values for thin and thick CVM were obtained to be 76.00 and 24.00, respectively. The per cent typical, atypical and nil fern patterns of CVM were obtained to be 91.23, 8.77 and 0.00; and 69.77, 30.23 and 0.00, respectively, in conceived and nonconceived cows, with the difference being statistically significant (P<0.05). The overall per cent values for typical, atypical and nil types of fern patterns of CVM were obtained to be 82.00, 18.00 and 0.00, respectively.. The mean spinnbarkeit values of CVM were found to be 14.61±0.33 (Range: 8.00-19.00) and 12.16±0.48 (Range: 7.00-18.00) cm, respectively, in conceived and non-conceived cows, with the difference observed statistically significant (P<0.01). Biochemical characteristics of CVM in conceived and non-conceived cows The mean pH value of CVM for conceived animals was found to be significantly (P<0.01) higher than that of non-conceived animals (7.57±0.05 Vs 7.31±0.05). The mean level of calcium (15.64±0.45 mg/dl) in CVM was obtained to be significantly (P<0.01) higher in non-conceived cows as compared to conceived cows (11.59±0.22 mg/dl). The mean inorganic phosphorus levels (mg/dl) in CVM of conceived cows was found to be significantly (P<0.05) higher as compared to that of the non-conceived cows (1.91±0.10 Vs 1.57±0.10). The mean magnesium concentration (mg/dl) in CVM for conceived animals was found to be lower than that of non-conceived animals (3.22±0.09 Vs 3.49±0.10). The mean sodium concentration (mEq/L) in CVM for conceived cows was obtained to be higher as compared to that of the non-conceived cows (167.70±7.78 Vs 162.15±8.03). The mean potassium concentration (mEq/L) in CVM for conceived cows was obtained to be higher as compared to that of the non-conceived cows (43.11±1.99 Vs 40.73±1.53). A significantly (P<0.01) higher mean concentration of total protein (4.23±0.04 g/dl) was obtained in CVM of conceived cows as compared to that of the pooled nonconceived cows (3.89±0.04 g/dl). Micro minerals Significantly (P<0.01) higher mean concentrations of copper, iron, zinc and manganese (1.31±0.07, 12.30±0.51, 1.16±0,04 and 2.03±0.12 ppm, respectively) in CVM of conceived cows were obtained as compared to that of the non-conceived cows (0.65±0.06, 7.16±0.37, 0.82±0.02 and 0.90±0.07 ppm, respectively). Body condition scores in conceived and non-conceived cows Out of 100 animals covered under the study, 57 (57.00%) conceived and 43 (43.00%) did not conceive. Among them, 41.28 per cent animals (n=57; 57.00%) were having 'good' body condition (BCS 3.0), followed by 'very good' body condition score (BCS 3.5) in 27 (27.00%) animals, 'fair' body condition score (BCS 2.5) in 10 (10.00%) animals and 'excellent' body condition (BCS 4.0) in 1 (1.00 %) animal. The maximum (62.96%) conception rate was obtained in animals having 3.5 (Very good) BCS, followed by per cent conception rates of 60.00, 56.14, 50.00 and nil in BCS groups of 2.0 (Poor), 3.0 (Good), 2.5 (Fair) and 4.0 (Excellent), respectively. BCS and physical characteristics of CVM in relation to fertility In the animals having BCS 2.0, the per cent incidences of clear colour, thin consistency, typical, fern pattern and spinnbarkeit value in CVM samples were found to be 100.00, 100.00, 100.00 and 15.00±0.57 cm; and 100.00, 100.00, 100.00 and 14.00 cm, in conceived and non-conceived cows, respectively. For the animals having BCS 2.5, 3.0 and 3.5, the per cent incidences of clear colour, thin consistency, typical fern pattern and mean spinnbarkeit (cm) values in CVM were found to be 80.00, 100.00, 100.00 and 13.80±0.66; 93.75, 87.50, 87.50 and 14.28±0.47; and 94.12, 88.24, 94.12 and 15.41±Q.78, respectively, in conceived cows. Among the cow (n=l) having BCS 4.0, none of the cows was conceived. BCS and biochemical characteristics of CVM of cows in relation to fertility The mean total protein levels (g/dl) in CVM of cows ranged from 3.84±0.05 to 4.31±0.10. The mean total protein levels in CVM of conceived cows were found to be slightly higher as compared to non-conceived cows under BCS 2.0, 2.5, 3.0 and 3.5 groups. The mean pH values of CVM of cows were found to be varying from 6.96±0.07 to 8.29±0.14. The mean pH values of CVM in 2.0 and 3.0 BCS were significantly (P<0.01) higher in conceived cows as compared to non-conceived (8.29±0.14 Vs 6.97±0.07 and 7.51±0.05 Vs 7.28±0.05, respectively). The similar trend was also observed in animals under 2.5 and 3.5 BCS groups, with the difference being nonsignificant. The mean calcium levels (mg/dl) in CVM of cows were ranged from 10.94±1.81 to 19.13±1.35. The mean calcium levels in CVM of conceived cows were found to be significantly (P<0.05) lower as compared to non-conceived cows under BCS 2.5, 3.0 and 3.5 groups. The mean inorganic phosphorus levels (mg/dl) in CVM of cows varied from 0.80±0.40 to 2.87. The mean inorganic phosphorus levels in CVM of conceived cows were found to be higher as compared to non-conceived cows under BCS 2.0, 2.5, 3.0 and 3.5groups, with the difference being non-significant. The mean magnesium levels (mg/dl) in CVM of cows ranged from 2.36 to 3.82±0.19. The mean magnesium level in CVM of conceived cows was found to be significantly (P<0.05) lower as compared to non-conceived cows under BCS 3.5. The similar trend was also observed in animals under 2.0, 2.5 and 3.0 BCS group, with the difference being non-significant. The mean sodium levels (mEq/L) in CVM of cows varied from 144.90±18.59 to 189.00±60.00. The mean sodium levels in CVM of conceived cows were found to be higher as compared to non-conceived cows under BCS 2.0, 2.5 and 3.0 gi'oups, whereas, the mean sodium levels (mEq/L) of CVM in 3.5 BCS was lower in conceived cows as compared to non-conceived (148.50±17.96 Vs 178.30±18.58), with the difference being non-significant. The mean potassium levels (mEq/L) in CVM of cows ranged from 32.50±6.72 to 45.50. The mean potassium levels in CVM of conceived cows were found to be lower as compared to non-conceived cows under BCS 2.0 and 3.5 groups, whereas, the mean potassium levels (mEq/L) of CVM in 2.5 and 3.0 BCS was lower in conceived cows as compared to non-conceived, with the difference being non-significant. BCS and trace elements in CVM of conceived and non-conceived animals The mean copper levels (ppm) in CVM of cows found to be varying from 0.32±0.31 to 1.34±0.11. The mean copper levels in CVM of conceived cows were found to be significantly (P<0.05) higher as compared to non-conceived cows under BCS 2.5, 3.0 and 3.5. The similar trend was also observed in animals under 2.0 BCS group, with the difference being non-significant. The mean iron levels (ppm) in CVM of cows varied from 5.11 to 12.59±0.78. The mean iron levels in CVM of conceived cows were found to be significantly (P<0.01) higher as compared to non-conceived cows under 3.0 and 3.5 BCS groups. The similar trend was also observed in animals under 2.0 and 2.5 BCS group, with the difference being non-significant. The mean zinc levels (ppm) in CVM of cows ranged from 0.71 ±0.13 to 1.28±0.11. The mean zinc levels in CVM of conceived cows were found to be significantly (P<0.05) higher as compared to non-conceived cows under BCS 2.5, 3.0 and 3.5 groups. The similar trend was also observed in animals under 2.0 BCS group, with the difference being non-significant. The mean manganese levels (ppm) in CVM of cows were found to be varying from 0.51±0.36 to 2.44±0.28. The mean manganese levels in CVM of conceived cows were found to be significantly higher (P<0.01) as compared to non-conceived cows under BCS 3.0 and 3.5. The similar trend was also observed in animals under 2.0 and 2.5 BCS groups, with the different obsei-ved to be non-significant.ThesisItem Open Access INDUCTION AND SYNCHRONIZATION OF ESTRUS IN POSTPATRTUM ANOESTRUS KANKREJ COWS(AAU, Anand, 2010) BHORANIYA, HARESHKUMAR LABHUBHAI; Dhami, A. J.This study was carried out at Livestock Research Station of the University on 36 Kankrej cows of 3 to 8 months postpartum period. The study included use of four estrus induction/synchronization protocols, viz. (i) Ovsynch (ii) CIDR (iii) Ovsynch + CIDR and (iv) Heatsynch on six anoestrus cows each, keeping 12 animals (6 cyclic-fertile and 6 anoestrus) as control. The effect of these hormone protocols was evaluated by comparing their estrus induction response and conception rates to fixed time inseminations as well as through monitoring plasma progesterone and other metabolites at different time interval (0,7, 9/11 -AI) after treatment and on day 20 post-insemination. The Ovsynch protocol was initiated in six Kankrej cows with postpartum anoestrus period of more than four months. All six (100 %) cows exhibited synchronized ovulatory estrus within 24 hrs of second GnRH injection with prominent, moderate and weak estrus signs in 2, 3 and 1 cows, respectively. The conception rates during the first, second and third service/cycle following induced estrus were 33.33, 25.00 and 0.00 per cent, respectively, with an overall conception rate of 50.00 (3/6) per cent. Palpation per rectum on day 60 post-insemination, revealed pregnancy rate of 50.00 per cent. The use of CIDR protocol in postpartum anoestrus Kankrej cows caused induction of ovulatory estrus in all six cows with prominent estrus signs in most of the animals. These animals were bred by FTAI and ultrasonographic scanning on day 40 revealed first, second and third service/cycle conception rate following induced estrus as 66.66, 50.00 and 100.00 per cent, respectively, with an overall conception rate of 100.00 (6/6) per cent. Palpation per rectum on day 60 post-AI indicated 83.33 % pregnancy rate, suggesting one case of embryonic mortality between days 40 and 60. A combined Ovsynch + CIDR protocol tested in six postpartum anoestrus Kankrej cows induced and synchronized estrus within 24 hrs of second GnRH injection in all the 6 animals with prominent, moderate and weak estrus signs in 3, 2 and 1 cows, respectively. The animals were bred by FTAI with the conception rates at induced, second and third service/cycle as 50.00, 0.00 and 33.33 per cent, respectively, with an overall conception rate of 66.66 (4/6) per cent. Palpation per rectum on day 60 post-AI also revealed pregnancy rate of 66.66 (4/6) per cent. The Heatsynch protocol tried in six postpartum anoestrus Kankrej cows induced estrus in all animals with prominent, moderate and weak estrus signs in 4, 2 and 0 cows, respectively. The conception rates following AI at first, second and third service/cycle were 16.66, 40.00 and 0.00 per cent, respectively, with an overall conception rate and pregnancy rate of 50.00 (3/6) per cent each. In the normal cyclic control group, all the six (100 %) cows showed prominent estrus signs, while in anoestrus control group, only one (16.67 %) cow expressed natural estrus with moderate signs 28 days later and conceived at second cycle. The conception rates at first, second and third cycle were 50.00, 66.66, 00.00 per cent, respectively, in normal cyclic (control) group with the overall pregnancy rate was 83.33 (5/6) per cent. Upon transrectal ultrasound scanning using 5 MHz linear transducer on day 26 post-AI, the embryo proper was observed in 11 out of 12 predicted pregnant cows, though embryonic vesicle was seen in all 12. Moreover, both embryonic vesicle and embryo proper were detected in all 15 predicted pregnant animals on day 40 postinsemination. The sensitivity of early pregnancy diagnosis through ultrasound scanning on days 26 and 40 was 85.71 and 93.75 per cent, and the specificity 75.00 and 100.00 per cent, respectively. The positive predictive values of early pregnancy diagnosis were 80.00 and 100.00 per cent by day 26 and 40, while the corresponding negative predictive values were 81.82 and 90.91 per cent. The overall diagnostic accuracy was found to be 80.77 and 96.15 per cent, respectively. The accuracy of early pregnancy diagnosis through plasma progesterone assay on day 20 post-AI was 76.00 per cent. Among the four treatment groups, the plasma progesterone levels increased in almost all the animals within 7 days of treatment and then decreased drastically around the day 9/11 (day of FTAI). Later, increased/high progesterone levels were detected on day 20 post-Al in conceived cows, but in non-conceived cows the levels were much lower or basal on that day. Statistically, the plasma progesterone (ng/ml) concentrations were significantly higher on day 7 in Ovsynch (5.727 ± 1.26), CIDR (4.37 ± 0.66), Ovsynch plus CIDR (3.55 ± 0.34) and Heatsynch (5.92 ± 1.11) protocols as compared to the corresponding values obtained on day 0, 9/11 (Al) and on day 20 post-AI for the same protocols. The overall mean plasma progesterone levels on the day of initiating treatment were 1.27 ± 0.56, 0.47 ± 0.09, 1.03 ± 0.32 and 0.58 ± 0.31 ng/ml for the four protocols, respectively, and 0.67 ± 0.33 ng/ml in anoestrus control group. Biochemical investigation revealed that plasma total protein, total cholesterol, calcium and inorganic phosphorus profiles were within normal physiological limits, and neither varied significantly between sampling days of different protocols nor between protocols on any of the days/periods studied. Results on the study in general revealed that among the four estrus induction/synchronization protocols tested, CIDR protocol was the best. Relatively poor results obtained in Heatsynch protocol was attributed to reduced (one half) dose of GnRH and very high potency of estradiol benzoate used instead of estradiol velarate, that probably induced overt estrus signs. The accuracy of early pregnancy diagnosis by day 40 using USG and by day 60 through per rectal palpation was almost similar.ThesisItem Open Access ESTRUS INDUCTION IN PUBERTAL ANESTRUS GIR HEIFERS USING DIFFERENT HORMONE PROTOCOLS AND EARLY PREGNANCY DIAGNOSIS BY TRANSRECTAL ULTRASONOGRAPHY(AAU, Anand, 2010) KHADE, NITIN BABAN; PATEL, D. M.The present study was carried out on 24 pubertal anestrus Gir heifers at Baroda District Co-op. Milk Producers Union Ltd., Baroda Dairy, Vadodara at their Heifer Development Project (Gir), Itola (Gujarat) during January 2010 to May 2010. The animals were divided into four groups (six in each group) and six animals were kept as control. The study included use of three hormone protocols: i) Ovsynch ii) Ovsynch plus CIDR and iii) CIDR alone. The effect of these hormone protocols was studied using different parameters and B-mode transrectal ultrasonography and the findings were evaluated by comparing them with plasma progesterone levels at different time intervals after treatment. Also, the arborization / fern pattern of estrus (induced) mucus was studied. The Ovsynch protocol was initiated in Gir cow heifers during puberty (> 2 yrs of age), considering the day of first GnRH injection as day 0. All the animals of this group showed induced estrus and ovulation. Estrus was detected between 10-20 hrs after the last GnRH injection with ovulation 9 to 15 hrs after the end of estrus in all the animals. The animals were bred by fixed-time AI (FTAI) twice (day 10) and ultrasonography on day 26 post-insemination revealed first service and second service conception rates as 16.66% and 40.00%, respectively, giving overall conception rate of 50.00%. Ultrasonography on day 40 and rectal palpation on day 60 postinsemination revealed both pooled conception rate and pregnancy rate of 50.00%. No early embryonic death (EED) or early fetal death (EFD) was noticed during the study. Combination of Ovsynch-CIDR protocol was initiated in pubertal anestrus animals, considering the first day of treatment as day 0. Induction of estrus and ovulation occurred in all the six animals and the animals were bred by FTAI and ultrasonography on day 26 post-insemination revealed first service conception rate, second service conception rate and overall conception rate of 50.00%) (three out of six animals), 33.33%) (one out of three animals) and 66.66% (four out of six animals), respectively. Both, ultrasound scanning and rectal palpation on day 40 and 60 postinsemination revealed pooled pregnancy rate of 66.66%o. The use of CIDR protocol in pubertal anestrus animals caused induction of estrus in all the six animals. These animals were bred by FTAI and ultrasonographic scanning on day 26 and 40 post-insemination revealed first service and second service conception rates were 33.33%) and 50.00%), respectively and pooled conception rate of 66.66%o (four out of six animals). The rectal palpation on day 60 post-insemination indicated 66.66% pregnancy rate. In the control group, all the six animals were anestrus as observed by rectal examination at regular interval of time. The plasma P4 levels in all these six animals remained < 1.20ng/ml throughout the research period. Upon ultrasound scanning, the embryo proper was observed on day 26 postinsemination. However, the embryo was observed in seven out of 11 heifers. The amniotic and allantoic vesicles were observed as non-echogenic cavities closely surrounding the embryo proper on day 40 post-insemination. These cavities were surrounded by large anechoic area i.e. embryonic fluid. The heartbeats were observed in 10 out of 13 animals on day 26 and in all 12 animals on day 40 post-insemination. The mean heart rates were found to be 174.60 ± 0.81 beats per minute on day 26 postinsemination, which later on decreased to 165.28 ± 1.4 beats per minute on day 40 post-insemination. The sensitivity of ultrasound scanning was cent percent on day 26 and 40 post-insemination. The specificity of ultrasound scanning was 87.50 per cent and 100 per cent on day 26 and 40 post-insemination, respectively. The positive predictive value (PPV) of the ultrasound scanning was 90.90 per cent and 100 per cent on day 26 and 40 post-insemination, respectively. The negative predictive value (NPV) of the ultrasound scanning was cent per cent on both the days i.e. day 26 and 40 post-insemination. The overall accuracy/diagnostic accuracy in the present study was recorded to be 94.44 per cent and 100 percent on day 26 and 40, respectively. All the 24 heifers included in this study were anestrus, having plasma progesterone levels ranged from 0.56-1.27ng/ml. After initiation of treatment, plasma progesterone levels increased in all the animals among the treatment groups upto 9 - 10 days and then decreased drastically around the day of estrus. Plasma progesterone concentrations in estrus heifers ranged from 0.31 - 0.53ng/ml (day 9, 10 and 11). Plasma progesterone concentrations ranged from 3.10 - 5.26ng/ml with an average of 4.04 ± 0.29ng/ml in pregnant animals on day 20 post-insemination. In non-pregnant animals plasma progesterone concentrations ranged from 1.20-2.10ng/ml with an average of 1.68±0.15ng/ml on day 20 post-insemination. The fern pattern of estrus mucus in animals induced to estrus was studied and it was found that conception rates were better in typical type of fern pattern when compared to atypical type. The difference was non significant. No conception was found in the nil type of fern pattern. These results suggest that estrus induction in pubertal anestrus Gir heifers using different hormone protocols viz. Ovsynch, Ovsynch and CIDR and CIDR alone are extremely useful in initiating cyclicity in pubertal anestrus/acyclic Gir heifers and may serve as an excellent tool for induction of estrus, ovulation and conception in pubertal anestrus Gir heifers. Ovsynch and CIDR combination as well as CIDR alone were found to be better protocols as pregnancy rates (66.66%) were good in Gir heifers. Ultrasonography facilitates diagnosis of all non-pregnant animals at an early stage (day 26 post-insemination) and is really advantageous than pregnancy diagnosis by rectal palpation method, with 100 per cent reliability. Thus, ultrasound scanning appeared to be of immense value in detection of early pregnancy, early nonpregnancy and early embryonic mortality.ThesisItem Open Access STUDIES ON PHYSICO-BIOCHEMICAL CHARACTERISTICS OF ESTRUAL CERVICO-VAGINAL MUCUS WITH REFERENCE TO BODY CONDITION SCORE AND FERTILITY IN GIR COWS(AAU, Anand, 2010) GOHEL, MUKESHBHAI MANUBHAI; Kavani, F. S.The present investigation was carried out during November 2009 to April 2010 on 50 Gir cows (10= normal cyclic, 20=repeat breeder and 20=induced estrus cows) from Livestock Research Station, AAU, Anand and Heifers Development Project of the Baroda District Co-operative Milk Producers' Union Ltd., Itola (Vadodara) as well as private farms of Anand district of Gujarat. Further all three groups were divided into two subgroups, conceived and non-conceived. The study included evaluation of physicobiochemical properties of estrual cervico-vaginal mucus (CVM) collected by standard procedure and their relationships with the body condition scores and fertility of cows. Among 10 normal cyclic cows, 70 and 30 per cent cows had clear and turbid mucus. Among 20 repeat breeding cows 20, 60 and 20 per cent cows had clear, turbid and dirty mucus, respectively and in induced estrus cows the figures were 50, 40 and 10 per cent. Overall, the frequency of clear, turbid and dirty colour mucus was 30, 24 and 4 per cent, respectively, in conceived cows and 12, 22 and 8 per cent in non-conceived cows. The conception rate was higher in normal cyclic cows (100 %) with clear cervical mucus compared to repeat breeder cows (50 %) and induced estrus cows (60 %). The dirty mucus was not observed in normal cyclic cows. Overall 75.00 and 41.67 per cent cows conceived with turbid mucus in induced estrus and repeat breeder groups, respectively. All the induced estrus cows with dirty mucus conceived, whereas none conceived in repeat breeder group with similar type of mucus. Out of 50 samples, 56 and 44 per cent were found to be thin and thick in consistency, respectively. The percentages of CVM samples with thin and thick consistency were 50-50 in repeat breeding, 55-45 in induced estrus cows and 70-30 in normal cyclic cows. Among normal cyclic cows, the frequency of the thin and thick mucus was 70 and 10 per cent, respectively, in conceived cows while only thick mucus (20 %) was observed in non-conceived cows. The frequency of thin and thick mucus for conceived and non-conceived repeat breeder cows was 20 and 15, and 30 and 35 per cent, respectively. The frequency of thin and thick mucus observed was equal for conceived induced estrus cows. The conception rate was higher in normal cyclic cows (100.00 %) with thin cervical mucus compared to repeat breeder cows (40.00 %) and induced estrus cows (63.64 %). The conception rate was higher with thick mucus (77.78 %) in induced estrus cows as compared to normal cyclic (33.33 %) and repeat breeder cows (30.00 %). The mean (±SE) pH of CVM in normal cyclic, repeat breeder and induced estrus cows was 7.17±0.04, 7.22±0.04 and 7.69±0.10, respectively. A higher mean pH of CVM was observed in non-conceived as compared to conceived cows of normal cyclic and repeat breeder groups, whereas in induced estrus group, pH of CVM was higher in conceived cows than non-conceived cows. Overall, non-significantly higher mean pH of cervico-vaginal mucus was observed in non-conceived (7.50±0.10) as compared to conceived cows (7.32±0.06). The mean (±SE) spinnbarkeit value of CVM in normal cyclic, repeat breeder and induced estrus cows was 14.40±0.30, 11.81±0.45 and 13.77±0.39 cm, respectively. Cows that conceived irrespective of normal or repeat breeder status had non-significantly higher spinnbarkeit values as compared to those that did not conceive whereas among induced estrus cows the spinnbarkeit value was higher in non-conceived as compared to conceived group. The atypical type of fern pattern was observed more in repeat breeders (55 %) and induced estrus cows (50 %) as compared to normal cyclic cows (20 %). A crystallization of cervico-vaginal mucus in induced estrus cows was typical and atypical in equal number (50 %). The degree of cervical mucus crystallization in the induced (CIDR) estrus group was lower than that in the normal estrus group. Among normal cyclic cows, 8 cows conceived with 70 and 10 per cent typical and atypical fern pattern, respectively. The frequency of typical and atypical fern pattern was 20 and 15 per cent in conceived and 20 and 45 per cent in non-conceived repeat breeder cows, whereas in the induced estrus cows the values were 30 and 40, and 20 and 10 per cent. The conception rate was higher in normal cyclic cows (87.50 %) with the typical fern pattern as compared to repeat breeder (44.44 %) and induced estrus cows (60.00 %) whereas, the higher conception rate was observed in induced estrus cows (80.00 %) with atypical fern pattern as compared to normal cyclic (50.00 %) and repeat breeder (27.27 %) cows. Nil type of fern pattern was not observed in any of the groups. A non-significantly higher mean total protein in CVM was recorded in pooled conceived as compared to non-conceived cows. A significantly (P<0.05) lower calcium was observed in cervico-vaginal mucus of normal cyclic cows (12.47±0.52 mg/dl) as compared to repeat breeder cows (15.44±0.62 mg/dl). The overall significantly (P<0.05) higher mean calcium (mg/dl) in CVM was observed in non-conceived (15.67±0.53) as compared to conceived cows (13.27±0.32). Contrary to calcium, significantly (P<0.01) lower mean inorganic phosphorus was observed in repeat breeders (1.26±0.14 mg/dl) as compared to normal cyclic (2.43±0.25 mg/dl) and induced estrus cows (2.29±0.25 mg/dl). Overall, significantly (P<0.05) higher mean value of inorganic phosphorus was observed in CVM of conceived (2.15±0.14 mg/dl) than the non-conceived cows (1.48±0.25 mg/dl). Non-significantly higher magnesium was observed in normal cyclic cows (4.34±.28 mg/dl) as compared to repeat breeder (4.25±0.17 mg/dl) and induced estrus cows (4.18±0.13 mg/dl). The mean magnesium in CVM of pooled conceived and nonconceived cows also did not differ significantly. Significantly higher mean values of sodium and potassium were recorded in CVM of conceived (181.01±4.85 and 32.42±0.81 mEq/L) as compared to non-conceived cows (130.61±7,66 and 25.17±0.94 mEq/L, respectively). The mean values of sodium and potassium were also higher in normal cyclic cows (169.69±7.73 and 31.91±0.91 mEq/L) and induced estrus cows (174.03±6.55 and 33.52±0.90 mEq/L) as compared to repeat breeders (140.75±10.35 and 24.27±0.68 mEq/L). Overall the means of copper, iron, zinc and manganese in cervico-vaginal mucus were found to be higher in pooled conceived cows as compared to non-conceived cows. A higher mean value of body condition score was observed in repeat breeders (3.27±0.07) as compared to induced estrus (3.00±0.07) and normal cyclic cows (2.90±0.08). Higher mean calcium was observed in CVM of repeat breeder cows that did not conceive than those conceived with 3.5 BCS. The mean sodium level was observed significantly higher in CVM of conceived as compared to non-conceived induced estrus cows with 2.5 BCS. Significantly (P<0.05) higher body condition score was observed in non-conceived (3.21 ±0.07) as compared to conceived cows (2.95±0.06). Thus the required number of Al/service per conception was more in cows with higher BCS. All the physical and certain biochemical properties of CVM were found to be independent of conceived and non-conceived status of normal cyclic, repeat breeder or induced estrus cows. However there were some positive associations of these traits with fertility rate.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.