H. P. Gupta

Effect of including growth factors and antioxidants in maturation medium used for in vitro culture of buffalo oocytes recovered in vivo.

This study examined the effect of including one of two growth factors (100 ng/ml IGF-1 or 20 ng/ml EGF) in combination with one of two antioxidants (50 microM cysteamine or 50 microM beta-mercaptoethanol) in maturation, fertilization and subsequent development of buffalo oocytes. The oocytes were recovered by in vivo ovum pick-up technique from six Murrah buffalo heifers twice a week over a period of 16 weeks. Immediately after ovum pick-up oocytes recovered from six donors were allocated randomly to five different maturation treatments. The control treatment was the basic maturation medium (MM; TCM-199 supplemented with 10% FBS, 10 IU/ml LH, 0.5 microg/ml FSH, 1 microg/ml estradiol-17beta and 50 microg/ml gentamicin). The other four treatments consisted of the control maturation medium (MM) plus one combination of a growth factor and an antioxidant viz. IGF-1+cysteamine; IGF-1+beta-ME; EGF+cysteamine or EGF+beta-ME. The total number of oocytes assigned to each maturation treatment ranged from 31 to 66. After maturation in different maturation medium, media used for in vitro fertilization and subsequent development of embryo was same for all groups. Data were analysed using Chi-square test. The maturation rate observed for the growth factor plus antioxidant treatments was similar to that for the control (90.4%). The highest cleavage rate recorded in the IGF-1+cysteamine treatment (71.9%) which was significantly higher (P<0.05) than the IGF-1+beta-ME (45.2%) and EGF+beta-ME (46.4%) treatments, but not significantly differ from the control (63.8%) and EGF+cysteamine treatment (60.7%). The proportion of cleaved oocytes those developed to blastocyst stage was significantly higher in the IGF-1+cysteamine treatment (52.2%; P<0.05) than in the control (23.3%), the EGF+cysteamine (13.5%) or the EGF+beta-ME (7.7%) treatments, but did not differ significantly from the IGF-1+beta-ME (28.6%) treatment. Following non-surgical transfer of 15 embryos to 14 synchronized recipients, four became pregnant and only one recipient sustained the pregnancy as long as 4.5 months when spontaneous abortion occurred. It was concluded that supplementing the maturation medium with IGF-1+cysteamine improved the production of buffalo embryos significantly in vitro culture.

Effect of heat stress on pregnancy rates of crossbred dairy cattle in Terai region of Uttarakhand, India

Abstract Objective To test whether heat stress during one or both of the critical periods (pre-insemination and post-insemination) causes reduction in pregnancy rates of crossbred dairy cattle in Terai region of Uttarakhand. Methods Data on metereological (ambient temperature and humidity) and reproductive (date of artificial insemination and day 60 reproductive status) variables were collected for a total of 1199 inseminations and allocated, on the basis of temperature humidity indices during 30 days before and after insemination, to one of the four groups: Thermoneutral-thermoneutral (TN-TN), Thermoneutral-heat stress (TN-HS), Heat stress-thermoneutral (HS-TN), and heat stress-heat stress (HS-HS). Results Chi -square analysis of the day 60 pregnancy data revealed lower ( P Conclusions These results indicate that reduction in pregnancy rates of crossbred dairy cattle due to heat stress in the Terai region was evident only when both growth of the ovulatory follicle and fertilization and early embryonic development occurred in the heat stress period. In addition, the study showed that use of temperature humidity indices rather than the traditional seasonal classification could provide more accurate insights into the effects of heat stress on pregnancy rates of crossbred dairy cattle in the Terai region.

Plasma progesterone profile and conception rate following exogenous supplementation of gonadotropin-releasing hormone, human chorionic gonadotropin, and progesterone releasing intra-vaginal device in repeat-breeder crossbred cows.

Aim: This study was designed to evaluate the effect of gonadotropin-releasing hormone (GnRH), human chorionic gonadotropin (hCG), and progesterone impregnated intra-vaginal device on progesterone profile and conception rate in repeat-breeding crossbred cows. Materials and Methods: Repeat-breeding crossbred cows aged 3-8 years (n=32), lactating and negative to white side test were randomly divided into four groups: Group 1 (Control, n=8), Group 2 (GnRH at 10 µg i.m, n=8), Group 3 (hCG at 1500 IU i.m., n=8), and Group 4 (progesterone impregnated intra-vaginal device at 958 mg, n=8). All the treatme nts were given on 5th daypostbreeding and in Group 4 intra-vaginally implanted device was withdrawn on 9th day (i.e., implant inserted for total 4 days) of the estrous cycle. Blood samples were collected on day 0, 5, 10, 15, and day 20 of estrous cycle, and plasma was separated for progesterone estimation. Results: Accessory corpus luteum was not formed in crossbred cows of Group4 and control group. However, total 6 and 8 accessory corpora lutea were found in Group 2 and Group 3, respectively. In pregnant cows, the plasma progesterone concentration increased continuously from day 0 to day 20. In non-pregnant cows, it increased from day 0 to day 15 and then declined. The conception rate on day 60 in Group 1, Group 2, Group 3, and Group 4 was 37.5%, 50%, 75%, and 37.5%, respectively. Conclusions: Treating repeat-breeder cows with hCG is effective in increasing conception rate by developing accessory corpora lutea and higher progesterone level.

Production of the first viable ovum pick–up and in vitro embryo produced (OPU–IVEP) buffalo calf in India

Abstract This report puts on record production of the first viable calf following ovum pick-up in vitro embryo production (OPU-IVEP) technique in buffalo ( Bubalus bubalis ) in India. A total of 313 transvaginal OPU sessions were performed in six elite Murrah buffaloes and 1 171 follicles were aspirated. From these follicles, a total of 729 immature oocytes were recovered. The recovered oocytes were graded as A (59, 8.1%), B (111, 15.2%), C (293, 40.2%) and D (262, 35.9%) following standard procedures. Maturable grade oocytes (A, B, and C; 473, 68.9%) were subjected to in vitro maturation in Medium 199 supplemented with 10% FBS, 0.5 μg/mL FSH, 10 IU/mL LH, 1 μ g/mL estradiol-17 β, 20 ng/mL EGF and 50 μM cysteamine in CO 2 incubator at 39 °C temperature, 5% CO 2 and high humidity. The matured oocytes (427 out of 473; 90.3%) were co-incubated with 1×10 6 /mL buffalo sperms in modified synthetic oviductal fluid (mSOF) containing 10 ng/mL heparin for 22 hrs for in vitro fertilization. After fertilization the presumptive zygotes were stripped off of remaining cumulus cells and cultured in mSOF for 7 days to study embryonic development. A total of 278 (65.1%) oocytes cleaved after 24-36 hrs of fertilization. Out of the cleaved oocytes, 249 (89.6%), 216 (77.7%), 167 (60.1%), 139 (50.0%) and 73 (26.3%) developed up to 4-cell, 8-cell, 16-cell, morula, and blastocyst stages, respectively. A total of 29 embryos were transferred into 27 synchronized recipient buffaloes. Two of these buffaloes were found pregnant at 90 days. One of them aborted at 4.5 months of gestation (male fetus) and the other gave birth to a live male calf weighing 25 kg. Currently, the calf is normal and has attained the age of 4.5 years.

Endometritis Increases Pro-inflammatory Cytokines in Follicular Fluid and Cervico-vaginal Mucus in the Buffalo Cow

ABSTRACT Emerging evidence shows that some of the pro-inflammatory cytokines are elevated not only in the endometrium but also in the follicular fluid of cows with endometritis. Developing a cervico-vaginal mucus (CVM) based test has the potential for becoming a pen-side test because of the ease of sample collection. The present study describes the results of two different experiments. The first experiment was conducted to investigate the influence of endometritis on the proinflammatory cytokines of follicular fluid based on the reproductive tracts of buffalo collected at a slaughter house Buffalo genitalia were categorized into purulent endometritis (PE), cytological endometritis (CE), and non-endometritis (NE) based on the white-side test and endometrial cytology, respectively (n = 14/group). Each group was subdivided into follicular and mid-luteal stage (n = 7/stage) and the follicular fluid was collected from the largest follicle. Second experiment was done to study the difference in the levels of proinflammatory cytokines in the CVM of repeat breeders with subclinical endometritis presented to the clinic. CVM was collected from the repeaters (n = 10) and non-repeaters (n = 10) through aseptic trans-vaginal aspiration. The pro-inflammatory cytokines such as IL-1β, IL-6, IL-8, and TNFα were quantitated through bovine specific ELISA kits. Significantly higher concentrations of pro-inflammatory cytokines (IL-1β, IL-8, IL-6, and TNFα) along with low intra-follicular estradiol in buffaloes of PE and CE groups suggest that endometritis impedes the follicular steroidogenesis. Significantly higher concentration of IL-1β and TNF-α in the CVM of repeaters indicate their potential as a pen-side diagnostic test for CE.

Fetal Dystocia due to Postural Defects of Neck and Carpals in a Jenny: Case Report

This article puts on record successful management of dystocia due to lateral deviation of head and neck and flexion of carpal of both forelimbs in a jenny. A 5 year old jenny in her first parity was presented to outpatient department of Veterinary Teaching Hospital, College of Veterinary and Animal Sciences, G.B. Pant University of Agriculture and Technology, Pantnagar with a history of complete gestation, straining since morning, the water bag ruptured and treated by a local vet without any success. Per vaginal examination revealed complete dilatation of cervix and dry foaling passage. The case was diagnosed as dystocia due to lateral deviation of head and neck and flexion of carpal of both forelimbs. The postural abnormalities of the fetus were corrected by applying repulsion on the brisket region and the head and neck were brought into normal posture by holding the mouth of the fetus in the palm. The fore limbs were brought to normal posture by grasping the hoof of the fetus and straightened one by one. Snares were applied on both the fore limbs at fetlock joint and a blunt eye hook was placed in the right orbit of the fetus. The fetus and the foaling passage were thoroughly lubricated with liquid paraffin and traction is applied. A dead male foal was delivered per vaginally after applying traction. Post operative medication of the jenny was done and healthy recovery was observed.

Trivers–Willard hypothesis revisited: Does heat stress peri–insemination alter secondary sex ratio in crossbred dairy cattle?

Abstract Objective To test the hypothesis that heat stress peri–insemination skews towards female the secondary sex ratio in dairy cattle. In addition, the effect of heat stress peri–insemination on birth weight of resultant calves was investigated. Methods Data on the date of insemination and sex and birth weight of the resultant calf were collected for a total of 934 single births on a crossbred dairy farm and grouped into thermoneutral and heat stress peri–insemination groups on the basis of temperature humidity indices on the day of insemination. Results Logistic regression revealed no difference in the secondary sex ratios between thermoneutral (53.4:46.6) and heat stress (52.5:47.5) peri–insemination groups. These sex ratios were not different from the expected 50:50 ratio on Chi–square goodness of fit test. Differences in birth weight of calves between thermoneutral and heat stress peri–insemination groups did not approach statistical significance. Conclusions These results indicate that heat stress peri–insemination does not affect secondary sex ratio and calf birth weight in crossbred dairy cattle.