Aim:The aim was to compare commercially available soybean milk-based extenders, viz. Bioxcell® and Optixcell® (IMV, France) with standard Tris-citrate-fructose-egg yolk-glycerol (TFYG) extender for cryopreservation of buffalo semen.Materials and Methods:Semen was collected twice a week in artificial vagina from six sexually mature, 4-6 years old, healthy breeding bulls of Surti buffalo breed. In all 48 qualifying ejaculates (8 per bull) having initial motility >70% were split into three equal aliquots and were diluted (at 34°C keeping 100×106 sperm ml−1) in TFYG, Bioxcell and Optixcell extenders. The French mini straws filled from each aliquot were gradually cooled to 4-5°C, equilibrated at 4°C for 4 h and frozen in liquid nitrogen 2 vapor using programmable biofreezer. Just before freezing (post-equilibration) and 24 h after frozen storage, the samples were evaluated for various sperm quality parameters using standard protocols. Frozen semen straws were thawed in a water bath at 37°C for 30 s. The post-thaw incubation survival (37°C for 1 h) was assessed through motility rating at 0, 30 and 60 min of incubation.Results:The mean percentages of prefreeze sperms in TFYG, Bioxcell and Optixcell extenders in terms of progressive motility (69.48±0.37, 68.02±0.49, 70.94±0.38), viability (79.21±0.39, 77.38±0.48, 81.58±0.38), total abnormalities (7.90±0.14, 8.60±0.16, 7.08±0.15), intact acrosome (89.54± 0.18, 88.58±0.22, 90.52±0.21) and hypoosmotic swelling (HOS) reactivity (67.96±0.32, 65.65±0.42, 70.23±0.37) varied significantly (p<0.05) between extenders. Similar pattern of significant (p<0.05) variations between these extenders for post-thaw sperm progressive motility (47.71±0.79, 44.38±0.85, 49.90±0.90), viability (57.19±0.79, 53.85±0.84, 59.67±0.91), total abnormalities (12.33±0.17, 12.75±0.21, 11.27±0.18), intact acrosome (76.83±0.23, 75.90± 0.27, 78.50±0.25) and HOS reactivity (45.02±0.84, 42.31±0.82, 47.81±0.90) was also observed for TFYG, Bioxcell and Optixcell extenders. The recently launched improved soybean milk-based extender Optixcell excelled the older Bioxcell extender and even standard TFYG in respect of some of the sperm quality parameters.Conclusion:The advantages of soy lecithin-based bovine semen extenders over egg yolk regarding sanitary issues are unquestionable but still egg yolk-based semen extenders are widely used because of the cost factor and good in vivo fertility results.

Aim:The aim of this study was to evaluate the influence of peripartum protein and minerals supplementation on plasma profile of steroid hormones, metabolites, and fertility in rural buffaloes.Material and Methods:A total of 85 advanced pregnant (~8 months) pluriparous buffaloes selected at farmers’ doorstep in three tribal villages of Middle Gujarat were randomly divided into two groups, viz., control (n=45) and nutrients treatment (40). The buffaloes of treatment group (n=40), in addition to farmers feeding schedule/control, received daily 1.5 kg compound concentrate mixture (22% CP) and 50 g of chelated ASMM for 2 months each pre- and post-partum. Further, 15 buffaloes, each of control and treatment group, were injected parentrally (deep i/m) with 5 ml of micro-minerals (each ml containing Se, Zn, Cu and Mn at 5, 40, 15 and 10 mg, respectively), twice 2 months before and on the day of calving, keeping rest of the animals (control, n=30 and treatment, n=25) as controls. Blood sampling was done on days −60, −30, −15, 0, 15, 30, 45, and 60 peripartum for estimation of plasma progesterone and estradiol by standard RIA techniques and other metabolites using assay kits on biochemistry analyzer. The puerperal events and postpartum fertility were monitored through history and by fortnightly palpation per rectum till day 45 and then again at 120 days postpartum for both the groups and subgroups.Results:The mean plasma progesterone concentrations in all groups declined significantly (p<0.05) from day 60 to day 15 prepartum, reached to the basal levels (<0.5 ng/ml) on the day of parturition, and subsequently, reduced nonsignificantly till day 15 postpartum and then showed a rising trend from day 30 to 60 postpartum with significantly higher values at day 45 and/or 60. The mean plasma estradiol values increased with approaching parturition and were at its peak on the day of calving (p<0.01). Thereafter, there was a rapid fall in the levels by day 15 and it remained low till day 45-60 postpartum. The blood glucose values showed an increasing trend with advancing gestation, reaching the highest on the day of calving, dropped significantly (p<0.01) within 15 days postpartum, and thereafter showed consistent values. The buffaloes supplemented with peripartum nutrients maintained significantly (p<0.05) higher blood glucose concentrations than the control during the peak lactation. The plasma protein levels varied significantly (p<0.05) between days within the group with the lowest values on the day of calving, as well as between groups with higher (p<0.05) values on day 30 and 60 postpartum in treated group. Micro-minerals injected did not reveal significant influence on steroid hormones, blood glucose, or plasma protein. The mean plasma total cholesterol was significantly lower (p<0.05) in treatment than the control group. The mean values in micro-minerals injected subgroup were higher than the non-injected control subgroup during postpartum phase. The mean plasma triglyceride values in the pregnant buffaloes under both the gr...

Advanced pregnant healthy HF crossbred cows (n=20) of 2-4 parity were equally divided in to control (routine farm feeding-RFF) and treatment/nutrients supplementation (RFF + bypass fat @ 100-200 g/h/d and ASMM @ 50 g/h/d) groups and were studied from 2 wks prepartum to 8 wks postpartum for plasma profile of steroid hormones and metabolites on days -14, -3, 0, +3, +14, +28 and +42 as well as for puerperal events and postpartum fertility. The mean plasma progesterone values were maximum (>6 ng/ml) on day 14 prepartum, which declined significantly (p<0.01) on day 3 prepartum reached to the basal levels (<1 ng/ml) on the day of calving, remained basal till day 14, and thereafter showed a rising trend on days 28 and 42 postpartum. The oestradiol-17 values were at its peak on the day of calving (p<0.01), showed a rapid fall by day 3 postpartum and remained low till recrudesce of follicular activity around day 35 postpartum. However, there were no statistical differences between the two groups in either of the hormones, except on day 42 postpartum. The levels of cortisol and PGF 2  Metabolites (PGFM) were 3-8 times higher on the day of parturition as compared to values at day 14 pre-and postpartum, and declined further till day 42 postpartum reaching to prepartum levels. The plasma cholesterol gradually decreased as parturition approached and increased in postpartum days to reach the highest value (p<0.01) at day 42. The nutrients supplemented cows had significantly (p<0.01) higher plasma cortisol, PGFM and cholesterol values than the control cows around parturition. However, no such variation was noted in plasma protein profile. The period of uterine involution in control and supplemented groups was identical (31.97±1.82 and 30.27±1.41 days), yet the cows in treatment group resumed estrous cycle earlier (38.00±1.95 vs 42.32±4.14 days, p<0.05) and had shorter service period (85.22±7.17 vs 100.67±5.60 days) with improved pregnancy rate (80 vs 60 %) as compared to those in control group. Thus, the peripartum nutrient supplementation in crossbred cows was beneficial and had positive effect on the postpartum fertility and plasma cortisol, PGFM and cholesterol profile.

. How to cite this article: Savalia KK, Dhami AJ, Hadiya KK, Patel KR, Sarvaiya NP (2014) Influence of controlled breeding techniques on fertility and plasma progesterone, protein and cholesterol profile in true anestrus and repeat breeding buffaloes, Veterinary World 7(9): 727-732. Abstract Aim:The aim was to evaluate the estrus response, conception rate and plasma profile of progesterone, protein and cholesterol following use of different hormonal protocols in anestrus and repeat breeding buffaloes. Materials and Methods:This study was carried out on 20 true anestrus, 20 repeat breeders, and 10 normal cyclic buffaloes. Ten anestrus buffaloes each were treated with standard controlled internal drug releasing (CIDR) i/vg device and Ovsynch (GPG) protocols with fix timed artificial insemination (FTAI), and blood samples were obtained on day 0, 7, 9/10 (AI) of treatment and day 21 post-AI. Ten repeat breeding buffaloes with mature mid-cycle palpable corpus luteum (CL) were treated with i/m injection of 25 mg prostaglandin F2α (PGF 2 α) with FTAI twice at 72 and 96 h later, whereas other ten repeat breeding buffaloes in standing estrus were inseminated with simultaneous i/m injection of buserelin acetate-gonadotropinreleasing hormone (GnRH) 20 μg. 10 buffaloes exhibiting first estrus within 90 days postpartum and inseminated without any treatment served as normal cyclic control. Blood samples were obtained on day of PG injection, day of AI and day 21 post-AI for estimation of plasma progesterone, protein, and cholesterol.Results: CIDR and Ovsynch protocols resulted in 100 and 80% induction of estrus with conception rates of 40 and 30% at induced estrus, respectively, in anestrus buffaloes. Mid-cycle PGF 2 α treatment resulted in 90% estrus induction and 40% conception rate at induced estrus, while Buserelin acetate-GnRH 20 μg injection at AI resulted in 30% conception rate in repeat breeders. In normal cyclic control group also, the first service conception rate was 30%. The mean plasma progesterone concentrations on day 0, 7, 9/10 (AI) of treatment and on day 21 post-AI were found to be significantly (p<0.05) different in both CIDR and Ovsynch protocols, being higher on day 7 (day of PG injection) and on day 21 post-AI than on day 0 and 9/10 (FTAI), which were near basal levels. The mean plasma progesterone level was significantly (p<0.01) higher on the day of initiation of mid-cycle PGF 2 α treatment (3.81±0.67 ng/ml) in a repeat breeding buffaloes suggesting luteal phase. The mean plasma P 4 levels on day 21 post-AI were significantly (p<0.01) higher than on the day of estrus in both repeat breeders and in normal cyclic controls. The plasma P 4 value on day 21 post-AI was significantly (p<0.01) higher in conceived than non-conceived buffaloes in all five groups. The mean plasma total cholesterol and total protein concentrations in anestrus and repeat breeding buffaloes under different treatments did not vary significantly between sampling days. However the cholesterol content was significantly (p<0.05) lower (79...

Aim:To evaluate estrus induction response and fertility including plasma progesterone and biochemical profile following use of three standard hormonal protocols in anestrus crossbred cows.Materials and Methods:The study was carried out on 40 true anestrus and 10 normal cyclic cows. 10 anestrus cows each were treated with standard intravaginal controlled internal drug release (CIDR) device, Ovsynch (GPG) protocol, and Norgestomet ear implant with fixed-time artificial insemination (FTAI). 10 anestrus cows were kept as untreated control while 10 cows exhibiting the first estrus within 90 days postpartum without any treatment served as normal cyclic control. Blood samples were obtained from treated cows on day 0, 7, 9 (AI) of treatment and day 21 post-AI, and from control groups on the day of AI and day 21 post-AI for estimation of plasma progesterone, protein, cholesterol, calcium, and inorganic phosphorus profile.Results:The use of CIDR, Ovsynch, and Norgestomet ear implant protocols resulted in 100% estrus induction with conception rates at induced estrus of 60%, 50%, and 50%, and the overall of three cycles as 80%, 80%, and 70%. In untreated anestrus control (n=10), only three cows exhibited spontaneous estrus within 90 days of follow-up and conceived giving the first service and overall conception rates of 66.66% and 30.00%, respectively. In normal cyclic control (n=10), the conception rates at first and overall of three cycles were 50% and 80%. The overall mean plasma progesterone (P4) concentrations in anestrus cows studied on day 0 (initiation), 7 (prostaglandin injection and/or removal of implant), 9 (FTAI) of treatment and on day 21 post-AI revealed that the values on day 7 and 21 were significantly (p<0.01) higher than other two periods in all three groups. The concentrations were significantly (p<0.05) higher in conceived than non-conceived group on day 21 post-AI in CIDR (4.36±0.12 vs. 1.65±0.82 ng/ml) and Ovsynch (4.85±0.62 vs. 1.59±0.34 ng/ml), but not in Norgestomet ear implant (4.50±0.53 vs. 3.02±1.15 ng/ml) or normal cyclic group (5.39±0.67 vs. 3.13±0.37 ng/ml). The cholesterol and protein levels were significantly higher, but not the calcium and phosphorus, in normal cyclic control than in anestrus groups. The influence of treatment days and pregnancy status was not significant for any of the biochemical constituents in any of the groups.Conclusion:Ovsynch and/or CIDR synchronization protocol can be effectively used to improve fertility up to 80% in anestrus cows, as compared to 30% in anestrus control, combined with plasma progesterone to delineate the reproductive status before and after treatment.

A study was undertaken on semen of three mature bulls each of Gir, Surti and Murrah breed to evaluate the comparative motion characteristics and kinematics of their fresh and frozen-thawed spermatozoa by Biovis CASA. The ejaculates (n= 24/breed) having >75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and were assessed. Amongst motility traits, the total motile, rapid progressive motile and slow progressive motile spermatozoa percentage decreased significantly by 23.08 - 30.09, 43.57 - 55.18, 9.12 - 22.75 %, respectively, plessthan0.01), while non-progressive motile sperm (4.78 - 21.48%) and immotile sperm (164.38 - 178.38 %) percentage increased significantly ( plessthan0.01) in frozen-thawed semen compared to that of fresh semen. The post-thaw quality of semen of all three breeds was in acceptable range. The mean values of sperm velocity/kinematic parameters observed in frozen-thawed semen of Gir, Surti and Murrah bulls, based on total motile sperms, viz., average path velocity, curvilinear velocity, straight line velocity, linearity, straightness, beat-cross frequency, amplitude of lateral head displacement, dancing frequency and dancing mean decreased significantly by 13.70 - 17.79; 9.76 - 12.95; 13.28 - 21.90; 7.28 - 9.68; 4.36 - 7.79; 15.56 - 25.15; 8.78 - 10.50; 6.16 - 18.67 and 12.98 - 15.96 %, respectively, as compared to that of their fresh semen samples. However, wobbling index remained almost same for both fresh and frozen semen. All motility traits differed but none of kinematics/velocity traits differed significantly between breeds/species. The values of all velocity parameters for progressive motile sperms were higher than total motile sperms in all three breeds. The effect of freezing-thawing on velocity and kinematic attributes was much less compared to absolute sperm motility, and both the fresh and frozen-thawed sperms behaved identically with respect to their velocity and kinematics. The rapid progressive motile sperm in both fresh (r=0.41 to 0.92) and frozen-thawed (r=044 to 0.88) semen had significant correlations with most of their velocity traits, and the later were significantly and positively or negatively inter-related among each other in semen of all three breeds. It was therefore concluded that cryopreservation process significantly reduces the motility and kinematics attributes of bovine spermatozoa and, CASA analysis of fresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/ kinematics of spermatozoa.

No abstract

Fertility improvement camps were organized in different villages of middle Gujarat covering 1481 breedable cattle and buffaloes with the history of impaired fertility. The reproductive status of animals was ascertained through gynaeco-clinical examination per rectum. Blood sampling (n=305) was done from representative groups of animals to know the nutritional status. All non-infectious infertile animals were dewormed using either Inj. Ivermectin 100 mg s/c or bolus Fenbendazole + Ivermectin 3.0 g orally. Anestrus animals (n=265) were then nutritionally supplemented either with chelated area specific mineral mixture (ASMM, 50-60 g/d/h) or multi-minerals boli - one on alternate day, with and without bypass fat (75-80 g/d/h) for one month along with an advice to improve feeding level, estrus detection and timely breeding. Revisits were made after 2.5-3.0 months to check the reproductive status. The mean blood glucose concentrations were significantly lower (p less than 0.05), while total cholesterol and zinc levels were higher (p less than 0.05) in anestrus than in subestrus and/or repeat breeding cows and buffaloes. Moreover, the plasma total cholesterol levels in buffaloes were just one half to those of cows. Further, the plasma total protein, calcium, phosphorus, magnesium, iron, copper and cobalt concentrations were found to be nonsignificantly higher in repeat breeders than in anestrus or subestrus cows and buffaloes. Calcium was deficient in some 20% animals and calcium: phosphorus ratio varied from 1:1 to 1.8:1 in animals under different areas of study indicating its role in causing infertility in dairy animals. Supplementation of ASMM, multi-mineral boli alone or in combination with bypass fat in pubertal /postpartum anestrus animals resulted in 34-56% estrus induction response and 33-50% conception rate, both being better with mineral plus bypass fat supplementation than minerals alone.