S. P. S. Ghuman

Impact of pre-ovulatory follicle diameter on plasma estradiol, subsequent luteal profiles and conception rate in buffalo (Bubalus bubalis)

The present study was designed to investigate the impact of pre-ovulatory follicle (POF) diameter on the day of estrus on plasma estradiol concentration, subsequent luteal profile (corpus luteum, CL, diameter and plasma progesterone concentration) and conception rate in buffaloes. Twenty-eight buffaloes were synchronized with synthetic analogue of prostaglandin F(2α) (PGF(2α)) administered 11 days apart. Transrectal ultrasonography and jugular vein blood sampling was carried out on the day of estrus and on days 0 (day of ovulation), 5, 12, 16 and 21 post-ovulation. Out of 28 buffaloes, 11 (39.3%) were diagnosed pregnant on day 40 post-ovulation. Retrospective analysis revealed that the buffaloes getting pregnant had larger (p<0.05) POF diameter. In fact, all the buffaloes (n=5/5) having POF diameter between >14 and 16 mm conceived, whereas, conception rate in buffaloes with POF diameter between >12 and ≤14 mm (n=6/17) or <12 mm (n=0/6) was 35.3% and 0.0%, respectively. A positive correlation (r=0.57, p<0.05) was observed between POF diameter and plasma estradiol concentration at estrus. On day 5 post-ovulation, luteal profile was positively correlated (CL: r=0.34, p<0.05; plasma progesterone concentration: r=0.27, p>0.05) with POF diameter. Further, on the same day, plasma progesterone concentration was positively correlated (r=0.47, p<0.05) with CL diameter, however, this correlation was absent (r=0.05, p>0.05) during the subsequent luteal phase. Nevertheless, the post-ovulation luteal profile of pregnant buffaloes was higher (p<0.05) compared to non-pregnant counterparts. In conclusion, the diameter of POF in buffaloes has positive impact on plasma estradiol concentration at estrus, post-ovulation luteal profile and conception rate. The diameter of CL can be used as an indicator of luteal function at early but not at mid or late luteal phase of estrus cycle in buffaloes.

Induction of Ovulation of Ovulatory Size Non‐Ovulatory Follicles and Initiation of Ovarian Cyclicity in Summer Anoestrous Buffalo Heifers (Bubalus bubalis) Using Melatonin Implants

This study was conducted on summer anoestrous buffalo heifers to monitor the efficacy of melatonin for induction of ovulation and ovarian cyclicity. During pre-treatment period of 24 days, the ovarian dynamics of five cycling and 10 summer anoestrous heifers was monitored on each alternate day using a transrectal ultrasound scanner. Thereafter, during treatment period, these 10 anoestrous heifers along with additional seven anoestrous heifers were randomly allocated into non-implanted (n = 5) and implanted (n = 12, one melatonin implant/50 kg, 18 mg melatonin/implant) group. Non-implanted heifers were monitored on each alternate day till the confirmation of second-ovulation in implanted heifers. Pre-treatment period revealed the presence of dominant follicles in anoestrous heifers which attained the diameter comparable with ovulatory follicles of cycling heifers but failed to ovulate and regressed. Between 6 and 36 days (15.3 +/- 2.9 days) post-treatment, all the implanted heifers (p < 0.05) exhibited ovulation of dominant follicles; however none of the non-implanted heifers ovulated during the corresponding period. The first-interovulatory period in implanted heifers ranged between 8 and 28 days (18.0 +/- 1.8 days). The implanted heifers with short (<or=16 days) interovulatory period had short-lived corpus luteum (CL) that had smaller diameter and secreted less progesterone (p < 0.05). The diameter of CL was large (p < 0.05) and plasma progesterone was high (p < 0.05) following second-ovulation compared with first-ovulation in implanted heifers. In conclusion, using melatonin implants, ovulatory size nonovulatory follicles observed in summer anoestrous buffalo heifers can be successfully ovulated to initiate ovarian cyclicity.

Effectiveness of melatonin and controlled internal drug release device treatment on reproductive performance of buffalo heifers during out-of-breeding season under tropical conditions.

Sixteen Murrah buffalo heifers, divided into control and treatment groups of eight animals each, were used to study the effect of melatonin and controlled internal drug release (CIDR) device treatment on the resumption of ovarian activity during out-of-breeding season (summer solstice). Treated group was implanted with melatonin (18 mg of melatonin per 50 kg of body weight) for 45 days and then heifers of both groups received CIDR for 9 days. All heifers received intramuscular 500 IU eCG on the day before CIDR removal and 10 μg GnRH on the day after CIDR withdrawal. All animals were subjected to estrus detection daily. Blood sampling in conjunction with transrectal ultrasonography were performed twice weekly to determine serum concentrations of melatonin, progesterone, LH, and antioxidant enzyme activities, as well as to monitor the ovarian follicular activity. Melatonin treatment resulted in an increase (P < 0.01) in serum melatonin and a decrease (P < 0.01) in serum progesterone and LH. In addition, melatonin had no significant effect on the frequency of LH pulses. Furthermore, melatonin treatment increased (P < 0.01) the diameter of the largest follicle and the number of large follicles between Days 0 and 35 of melatonin treatment. However, melatonin exhibited superior ability to maintain CL at 21 days after artificial insemenation (AI) and increased the percentage of conception to threefold higher than control. In conclusion, melatonin implantation successfully improved the diameter of largest follicles and the ability to maintain CL at 21 days after AI in buffalo heifers during out-of-breeding season under tropical conditions.

Impact of buserelin acetate or hCG administration on day 12 post-ovulation on subsequent luteal profile and conception rate in buffalo (Bubalus bubalis)

The present study investigated the impact of gonadotropic hormone administration on day 12 post-ovulation on subsequent luteal profile and conception rate in buffaloes. All the buffaloes (n=48) were estrus synchronized by a synthetic analogue of prostaglandin F(2α) (PGF(2α)), administered 11 days apart, followed by insemination during mid to late estrus. To examine the effect of mid-luteal phase hormonal treatment, buffaloes were randomly divided into control (normal saline, n=14), d12-BA (buserelin acetate, 20μg, n=17) and d12-hCG (hCG, 3000IU, n=17) groups. Ovaries were scanned on the day of induced estrus to measure the preovulatory follicle (POF) diameter and on days 5, 12, 16 and 21 post-ovulation to examine the alterations in corpus luteum (CL) diameter. On the day of each sonography, blood samples were collected for the estimation of plasma progesterone. In treatment groups, luteal profile (CL diameter and plasma progesterone) on day 16-21 post-ovulation was better (P<0.05) as well as first service conception rate was higher (52.9% in each treatment group vs. 28.6%, P>0.05) compared to controls. All the pregnant buffaloes exhibited higher (P<0.05) plasma progesterone on various post-ovulation days than their respective non-pregnant counterparts. Treatment-induced accessory corpus luteum (ACL) formation was observed in 58.8 per cent and 70.6 per cent buffaloes of d12-BA and d12-hCG group, respectively, that also had higher (P<0.05) plasma progesterone compared to controls. Compared to the spontaneous CL, the diameter of ACL was less (P<0.05) in the treatment groups. In conclusion, buserelin acetate and hCG administration on day 12 post-ovulation leads to accessory CL formation, improves luteal profile and consequently increases conception rate in buffaloes.

Attenuation of luteolytic response following fish meal supplementation in dairy buffaloes (Bubalus bubalis)

Luteolysis of corpus luteum, due to un-inhibited PGF(2α) secretion, has been reported to be a cause of early embryonic mortality in dairy animals. The objective of this study was to determine the effects of fish meal (FM) supplementation on the uterine secretion of PGF(2α) and hence establish its supplementation as an antiluteolytic strategy in dairy buffaloes. Five cycling Murrah buffaloes were supplemented with 250g FM daily for 55 days in addition to their routine feed and seven buffaloes were kept as non-supplemented control. After 30 days of FM supplementation, the oestrus was synchronized in all the buffaloes using Ovsynch protocol. On day 15 of synchronized cycle, animals were challenged with oxytocin (OT; 100IU) intravenously and blood samples were collected at 15min interval, 1h before to 4h after OT challenge. The PGF(2α) response was measured as the venous concentration of 13,14-dihydro-15-keto PGF(2α) (PGFM). The mean hourly concentration of PGFM in FM supplemented buffaloes was lower than in the control buffaloes at all the occasions. During peak response (1h post-OT challenge), PGFM concentration was significantly lower (P<0.05) in FM supplemented buffaloes than in the control (197.4±41.7pg/ml versus 326.3±33.5pg/ml, respectively). Also the percent rise in PGFM after OT-challenge in FM supplemented buffaloes was less than the control (11.73% versus 22.47%). The dietary supplementation did not affect the size of corpus luteum (CL) and plasma progesterone concentration. Plasma glucose and total protein concentrations remained within the normal physiological limits during FM supplementation. The present study indicated that supplementing FM decreased the concentrations of PGF(2α) without alterations in the size of CL and plasma progesterone concentrations in dairy buffaloes.

Detection of estrous behavior in buffalo heifers by radiotelemetry following PGF2α administration during the early or late luteal phase.

This study examined the usefulness of radiotelemetry for estrous detection in buffalo heifers and the impact of prostaglandin F2α (PGF2α) administration during the early or late luteal phase on estrous behavior and ovulatory follicle variables. Induction of estrus with PGF2α at a random stage of the estrous cycle was followed by the arbitrary division of heifers into groups receiving a second dose of PGF2α during either the early (n=33) or late (n=17) luteal phase (6-9 or 11-14 days after estrus, respectively) for the induction of synchronized estrus. The electronic detection of synchronized estrus by radiotelemetry was confirmed using ultrasonography every 6h until ovulation. Radiotelemetry was 90% efficient and 100% accurate for estrous detection. Intervals between the PGF2α dose and the beginning of synchronized estrus (40.7 ± 10.9 vs. 56.7 ± 12.8h) or ovulation (70.0 ± 11.3 vs. 85.6 ± 12.5h) were shorter (P<0.05) for heifers receiving PGF2α during the early luteal phase. PGF2α administration during the early or late luteal phase produced similar (P>0.05) results for the duration of estrus, the intervals from the beginning or end of estrus to ovulation, the number and duration of mounts per estrus, the duration of mounts, the diameter of the ovulatory follicle and the luteal profile on day 5 after estrus. In conclusion, radiotelemetry was a suitable tool for the efficient and accurate detection of estrus in buffalo heifers. Treatment with PGF2α during the early luteal phase had a shorter interval to synchronized estrus and ovulation; however, estrous behavior, ovulatory follicle dynamics and subsequent luteal activity were similar following PGF2α administration during the early or late luteal phase.

Improvement of conception rate in postpartum flaxseed supplemented buffalo with Ovsynch + CIDR protocol

The present study was conducted on lactating Murrah buffalo to assess the effect of crushed flaxseed (a source of omega-3 fatty acids) supplementation (300g/100kg bwt/day for 60 days), over and above the routine feed, on luteolytic signal (PGF2α), luteal function (progesterone) and conception rate. In first experiment, on day 50 post-calving, six non-supplemented buffalo were treated to synchronize time of ovulation using an Ovsynch+Controlled Internal Drug Release (CIDR) protocol followed by intravenous oxytocin treatment (OT; 100IU) on day 15 post-ovulation. Blood samples were collected at 15min interval, 1h before to 4h after OT challenge. Thereafter, the same buffalo were supplemented with flaxseed, treated to synchronize time of ovulation starting on day 35 post-supplementation using the same protocol and subjected to OT treatment and blood sampling on day 15 post-ovulation. The PGF2α response was measured as the venous concentration of 13,14-dihydro-15-keto PGF2α (PGFM). The mean hourly concentration of PGFM subsequent to flaxseed supplemented was less (P<0.05) than in the pre-supplementation period at all the occasions. Flaxseed supplementation did not affect plasma fatty acids and other plasma metabolites except for an increase (P<0.05) in plasma cholesterol and plasma alanine transaminase. In the second experiment, 31 buffalo were randomly assigned to a control (n=16) and flaxseed supplemented (n=15) group. The latter group was supplemented with flaxseed starting from day 15 post-calving. On day 50-post-calving, buffalo of both groups were treated to synchronize time of ovulation among animals as described for the first experiment followed by artificial insemination (AI). Post-AI luteal phase plasma progesterone was greater (P<0.05) in the supplemented group compared to controls. Conception rate on day 63 post-AI was 66.7% in supplemented and 31.2% in controls (P<0.05). The present study indicated the beneficial impact of dietary supplementation of crushed flaxseed on conception rate through attenuation of luteolytic signal and improvement in post-breeding luteal profile.

Impact of buserelin acetate or hCG administration on day 5 post-ovulation on subsequent luteal profile and conception rate in Murrah buffalo (Bubalus bubalis)

The present study aimed to establish the impact of buserelin acetate or hCG administration on day 5 post-ovulation on subsequent luteal profile and conception rate in buffalo. The buffalo (n=45) were subjected to an estrous synchronization protocol (synthetic analog of PGF2α administered, through intramuscular route, 11 days apart), followed by artificial insemination (AI) during mid to late estrus. On day 5 post-ovulation, buffalo were administered (i.m.) normal saline (Control, n=14), buserelin acetate (20μg, d5-BA, n=14) or human chorionic gonadotropin (3000IU, d5-hCG, n=17). Ovarian ultrasonography was conducted on the day of induced estrus and on days 0, 5, 12, 16 and 21 post-ovulation to assess preovulatory follicle or corpus luteum (CL) diameter. Also, on these days, jugular vein blood sampling was conducted for the estimation of plasma progesterone. First service conception rate was greater (χ(2)=5.18, P>0.05) in d5-BA and d5-hCG groups (71.4% and 47.1%, respectively) as compared to control (28.6%). Both treatment groups had a greater (P<0.05) CL diameter and plasma progesterone during the post-treatment period in comparison to that control treatment group. Treatment-induced accessory CL formation was observed in 92.9% and 76.5% buffalo of d5-BA and d5-hCG groups, respectively. In conclusion, buserelin acetate and hCG administration on day 5 post-ovulation leads to accessory CL formation that may have a role in enhancing conception rate.