Exogenous progesterone-dependent modulation in the follicular dynamics of Bos indicus cattle undergoing repeated ovum pick-up sessions.

The objective of the present study was to evaluate the role of serum progesterone (P4) in follicular dynamics, oocytes' recovery and quality and their in vitro developmental competence during consecutive ovum pick-up (OPU) sessions in Bos indicus dairy cows. Wave-synchronized Sahiwal cattle (n = 20) were randomly divided into treatment (n = 10) and control (n = 10) groups. CIDR was used as a source of external progesterone in the treatment group. Four consecutive OPU sessions at 96-hr intervals were conducted and repeated ultrasonography at 12-hr intervals was done to monitor follicular dynamics. The viable oocytes were processed for IVC following IVM and IVF until day 7. The serum P4 concentrations in the P4 and control groups were recorded as 2.31 ± 0.059 versus.0.32 ± 0.065 ng/ml, respectively (p < .05). In the treatment group, the total number of recorded follicles was higher (p < .05; 12.05 ± 0.37 versus. 10.87 ± 0.40), whilst the growth rate (mm/day) of follicles was lower (p > .05). Per session recovered oocytes (5.31 ± 0.19 versus. 3.58 ± 0.21; p < .0001) and recovery rate (54.23 versus. 42.53%; p < .05) were also higher in the treatment group compared to control. Similarly, the viable oocytes (4.54 ± 0.187 versus. 3.06 ± 0.199) and the number of grade I and II oocytes per session (3.37 ± 0.196 versus. 2.06 ± 0.21) were higher (p < .05) in the treatment group compared with the control group. However, the nuclear maturation, cleavage, and blastocyst rate did not differ (p > .05) between the groups. Taken together, during OPU sessions, serum P4 improves oocytes' recovery and quality, whilst does not affect the in vitro developmental competence of recovered oocytes.

Effect of exogenous administration of oxytocin on postpartum follicular dynamics, oestrous rate and ovulation in Nili-Ravi buffaloes.

Based on different surveys, dairy farmers are concerned about extensive use of exogenous oxytocin in buffaloes, which is being held responsible for reproductive problems including irregular oestrous cycle and delayed ovulation. For these concerns, effects of oxytocin injection on postpartum follicular dynamics, postpartum oestrous interval (PEI), oestrous length, the interval from onset of estrus to ovulation and blood progesterone (P4) were studied in Nili-Ravi buffaloes. For this purpose, 23 animals within 1 week after calving were randomly divided into three groups: without oxytocin (CON; n = 7), 10 i.u. oxytocin (LOW; n = 8), 30 i.u. oxytocin - (HIGH; n = 8) and used to record the PEI for the study period of 154 days. At subsequent estrus, three buffaloes from each group (not served) were selected randomly to monitor two cycles for 6 weeks. Transrectal ultrasonography was performed to evaluate follicular and corpus luteum (CL) development, and blood sampling was done for progesterone (P4) analysis. These results revealed that postpartum oestrous interval (PEI) decreased significantly in oxytocin-treated groups. The number of small, medium and total follicles on the left ovary was significantly higher in the HIGH group. However, an overall number of small and total follicles on both right and left ovaries was significantly higher in CON and HIGH groups. On the other hand, there was no difference in the number of follicles on the right ovary among all treatment groups. The same was true for the size of pre-ovulatory follicles, CL, P4 concentrations and oestrous cycle length. The intervals from onset of estrus to ovulation and from standing estrus to ovulation were increased considerably in the HIGH group. It is concluded that exogenous oxytocin administration resulted in the shortening of PEI but triggered a delay in ovulation. Moreover, a higher dose of oxytocin could stimulate the growth of small, medium, and total follicles in postpartum Nili-Ravi buffaloes.

Effect of plasma progesterone on oocyte recovery, oocyte quality, and early in-vitro developmental competence of embryos in Bos indicus dairy cows.

The objective of present study was to determine the effect of plasma progesterone (P4) on oocyte recovery, oocyte quality, and early in-vitro developmental competence of embryos in Bos indicus dairy cows. The ovaries were collected in an abattoir. These ovaries (n = 750) were divided into two groups: 1) estrous CYCLIC (n = 318), and 2) estrous ACYCLIC (n = 432). Mean serum concentrations of P4 in a subset of (n = 85; 4.21 ± 0.4 ng/ml compared with 0.5 ± 0.2 ng/ml; P < 0.05) were greater in estrous CYCLIC as compared to ACYCLIC cows, respectively. The mean number of oocytes recovered per ovary (6.5 ± 0.5 compared with 4.0 ± 0.2; P < 0.05) was greater for estrous CYCLIC than ACYCLIC cows, respectively. The oocytes with grade I_+_II quality (55.3% compared with 47.6%; P < 0.05) were greater, whereas, there was lesser percentage with grade III_+_IV quality (44.5% compared with 52.4%; P < 0.05) from estrous CYCLIC as compared with ACYCLIC cows, respectively. Cleavage rate (70.9% compared with 52.8%; P < 0.05) was greater for embryos derived from estrous CYCLIC than ACYCLIC cows, respectively. Similarly, the embryo developmental rates to the 8- (38.5% compared with 20.8%; P < 0.05) and 16- (20.0% compared with 10.9%; P < 0.05) cell stage were greater for embryos derived from estrous CYCLIC as compared to ACYCLIC cows, respectively. In conclusion, the presence of greater plasma P4 has a beneficial effect on oocyte recovery, oocyte quality, and early IVEP outcomes in Bos indicus dairy cows.

Luteal blood flow measured by Doppler ultrasonography during the first three weeks after artificial insemination in pregnant and non-pregnant Bos indicus dairy cows.

The objective of this study was to determine if there are differences in luteal size (LS), progesterone (P4), and luteal blood flow (LBF) between pregnant and non-pregnant Bos indicus dairy cows during the first three weeks after insemination, and whether these parameters are related to each other. Lactating cows (n = 13) of mixed parity with a body weight of 430 ± 18 kg (mean ± SD), showing regular estrous cycle were used in the study. All cows were artificially inseminated and were classified as pregnant (embryonic heartbeat on day 30; n = 8) or non-pregnant (inter-estrus interval 17 to 21 days, n = 5). In order to compare the LS and LBF after artificial insemination, B-mode and color Doppler ultrasonography of ovaries were performed on days 4, 5, 6, 7 (first week), 8, 10, 12, 14, (second week), and 16, 17, 18, 19, 20, 21 (third week) in pregnant and non-pregnant cows. Results revealed that the mean LBF was consistently higher (P < 0.05) during days 7 through 21 in pregnant cows than in non-pregnant cows. The mean LS was higher (P < 0.05) on days 6 and 7, and from day 17 onwards, and the mean concentration of P4 was higher (P < 0.05) on days 19, 20, and 21 in pregnant cows. In conclusion, LBF is a more sensitive parameter than LS and P4 for detection of differences in luteal function between pregnant and non-pregnant Bos indicus dairy cows during the first three weeks after AI.

Equine chorionic gonadotropin (eCG) enhances reproductive responses in CIDR-EB treated lactating anovular Nili-Ravi buffalo during the breeding season.

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) on the estrous response, ovulatory follicle development, ovulation rate, corpus luteum (CL) diameter, concentrations of progesterone (P4) and pregnancy per AI (P/AI) in CIDR-EB treated lactating anovular Nili-Ravi buffalo during the breeding season. Buffalo (n = 87), 60 to 200 Days postpartum were treated with CIDR at a random stage of the estrous cycle (Day 0) and assigned to two groups; with eCG (+eCG; n = 44), or without eCG (-eCG; n = 43). In both groups, each animal received 2 and 1 mg estradiol benzoate (EB) on Day 0 and then on Day 8, respectively. On Day 6, eCG and PGF2α were administered. On Day 7, CIDR were removed and FTAI was performed at 48 and 60 h later and pregnancy was diagnosed 35 Days post AI. The results revealed that eCG treatment increased (P <  0.05) the CL diameter on 15, 18, and 21 after the ovulation that resulted from imposing the estrous synchronization protocol than when eCG was not administered. Similarly, treatment with eCG resulted in an increased (P <  0.05) mean concentration of P4 as compared with animals not treated with eCG on Days 18 and 21 after the ovulation that occurred subsequent to imposing the estrous synchronization protocol. The ovulation rate was greater in eCG-treated buffalo as compared with those not treated with eCG (93% compared with 74%, respectively; P < 0.05). The P/AI was greater in buffalo treated with eCG than those not treated with eCG (56% compared with 31%, respectively; P < 0.05). In conclusion, the addition of eCG along with EB and P4 based treatments enhanced the estrous response, follicular and luteal function, ovulation rate, concentrations of P4, and P/AI in lactating anovular Nili-Ravi buffalo.

Effect of resynchronization with GnRH or progesterone (P4) intravaginal device (CIDR) on Day 23 after timed artificial insemination on cumulative pregnancy and embryonic losses in CIDR-GnRH synchronized Nili-Ravi buffaloes.

The objective of the present study was to determine the effect of resynchronization on Day 23 with either GnRH or P4 (controlled internal drug release device containing progesterone; CIDR) on pregnancy rate, cumulative pregnancy, and embryonic and fetal losses in CIDR-GnRH synchronized Nili-Ravi buffaloes. Buffaloes (n = 181) of mixed parity, lactating, 181 ± 73 days postpartum, a body condition score (BCS) of 3.2 ± 0.5 (scale of 1-5), and 450-600 kg weight were subjected to synchronization and resynchronization. All buffaloes received CIDR on Day -9.5. In addition, GnRH was injected 36 h after CIDR removal, and timed artificial insemination (TAI) was performed 18 h later (Day 0). On Day 23, buffaloes were randomly assigned to receive one of the following treatments: 1) CON (n = 63), 2) P4 (n = 55), and 3) GnRH (n = 63) for resynchronization (2nd AI). Pregnancy rate, and embryonic and fetal losses were monitored by serial ultrasonography on Days 30, 45, 60, and 90 after synchronization (1st TAI), respectively. The pregnancy rate in GnRH-treated buffaloes remained significantly and consistently higher (P < 0.05) than in the CON group at Days 30, 45, 60, and 90 after 1st TAI. Based on the pregnancy diagnosis, on Day 30 post 1st TAI, buffaloes that remained non-pregnant in the CON, P4, and GnRH groups received: 1) Artificial insemination on detected estrus (AIDE; n = 37), 2) CIDR-GnRH protocol (CIDR; n = 27), and 3) Ovsynch protocol (OVS; n = 23), respectively. The pregnancy rate in resynchronized buffaloes did not differ (P > 0.05) between the OVS and CIDR groups; whereas the, cumulative pregnancy rate in GnRH + OVS buffaloes (81%) after 1st and 2ndAI when determined on Day 64 was higher (P < 0.05) than that in CON + AIDE (59%) buffaloes. The embryonic losses were significantly lower (P < 0.05) in GnRH-treated (18%) buffaloes, than in CON (42%) buffaloes on Day 45 post 1st TAI. Fetal losses were fewer and did not differ (P > 0.05) due to treatments on Day 60 or 90 post 1st AI. In conclusion, 1) the pregnancy rate and cumulative pregnancy rate in GnRH-treated buffaloes were higher than in CON buffaloes on Day 64 after synchronization and resynchronization, and 2) embryonic and fetal losses were lower in GnRH-treated buffaloes than in CON buffaloes when determined from Day 31-90 post 1st TAI.

Incorporation of estradiol benzoate to CIDR protocol improves the reproductive responses in crossbred dairy heifers.

The present study was designed to determine the effect of estradiol benzoate (EB) on reproductive response following a controlled internal drug release (CIDR) protocol in crossbred (Sahiwal × Friesian) dairy heifers. In the first trial, a total of 100 crossbred dairy heifers were treated with CIDR protocol for 7 days and injected with the PGF2α on day 6. After 24 h of CIDR removal, one group (EB = 50) was injected with estradiol benzoate whereas the other (control = 50) remained untreated. Estrus intensity and response were recorded visually and ovulation rate was recorded by ultrasonography. All heifers were artificially inseminated at 48 and 60 h following CIDR removal. Heifers were scanned for pregnancy within days 30-40 of artificial insemination (AI). In the second trial, two subgroups of heifers were included to observe the estrus and ovulatory events. The results of the first trial revealed that estrus response was achieved 100% in both the treatment groups. Estrus intensity (2.9 ± 0.1 vs. 2.0 ± 0.7) and ovulation rate (100 vs. 88%) differed significantly (P < 0.05) between the EB and control groups. However, a tendency for higher pregnancy per AI was observed (54 vs. 36%; P = 0.07) in EB than that in control groups. The results of the second trial revealed that a significantly (P < 0.05) shorter estrus and earlier ovulatory events were observed in EB-treated heifers. It is concluded that the incorporation of estradiol benzoate to the CIDR protocol is helpful to improve the estrus signs and enhance the ovulation and the pregnancy per AI in crossbred dairy heifers.

Effect of ovsynch versus prostaglandin F2α protocol on estrus response, ovulation rate, timing of ovulation and pregnancy per artificial insemination in Sahiwal cows.

Sahiwal cow is Bos indicus which is an important dairy breed of tropical and sub-tropical region. Research on reproduction is rare in this breed. The objectives of the present study were to determine the effect of Ovsynch (OVS) versus prostaglandin F2α (PG) protocol on estrus response and its intensity, ovulation rate, timing of ovulation and pregnancy per artificial insemination (AI) in Sahiwal cows. Experimental cows (n = 80) were of mixed parity, lactating, suckled, ≥120 days postpartum with body condition score 3.08 ± 0.34 and 375-475 kg of body weight which were randomly assigned to receive either OVS (n = 46) or PG (n =34) protocol. Cows were inseminated twice at 12 and 24 h after second gonadotropin releasing hormone in the OVS group, and 72 and 84 h after administration of prostaglandin F2α in PG group, respectively. The results revealed that estrus response did not differ (P > 0.05) and was 87% in OVS and 78% in PG cows. Ovulation rate did not differ (P > 0.05) and was 50% in both, OVS and PG cows. The pregnancy per AI did not differ (P > 0.05) and was 43% in OVS compared to 31% in PG cows. It is concluded that estrus response, ovulation rate and pregnancy per AI of OVS protocol is the same as PG in Sahiwal cows.

Presynchronization of lactating dairy cows with PGF2α and GnRH simultaneously, 7 days before Ovsynch have similar outcomes compared to G6G.

The overarching objective of this study was to develop an alternative strategy for first and greater services that will improve fertility in lactating dairy cows for dairy operations limited by labor or other logistical constraints that make it difficult to use Presynch-11, G6G, or Double-Ovsynch. Our overall hypothesis was that simplification of a Presynch program through the combination of PGF2α and GnRH on the same day (PG + G), 7 days before the first GnRH of Ovsynch, would allow for similar ovulation and luteolysis rate and pregnancies per AI (P/AI) compared with G6G. Lactating dairy cows 58 to 64 days in milk (first service; n = 114), and cows diagnosed not pregnant 39 days after previous AI (second + service; n = 122) were blocked by parity and service and randomly assigned to control or PG + G. Control cows received G6G (n = 116) that consisted of PGF2α, 2-day GnRH, 6-day GnRH, 7-day PGF2α, 56-hour GnRH, and 16-hour AI. Treated cows (PG + G; n = 121) received PGF2α and GnRH, 7-day GnRH, 7-day PGF2α, 56-hour GnRH, and 16-hour AI. All cows received a second PGF2α 24 hours after the PGF2α of Ovsynch. First service cows received AI between 76 and 82 days in milk and second + service received AI 57 days after previous AI. Pregnancies/AI (n = 230) were similar in controls compared with treated cows on Day 35 (57 vs. 50%; P = 0.27) and Day 49 (54 vs. 47%; P = 0.33), respectively. Percent of cows ovulating after GnRH of the presynchronization was greater (P = 0.002) for controls vs. treated (80 vs. 58%); however, ovulation after first GnRH of Ovsynch was similar (67 vs. 68%; P = 0.86). Serum concentrations of progesterone were similar (P = 0.78) at the time of first GnRH of Ovsynch for control and treated cows (2.22 vs. 2.14 ng/mL). However, serum progesterone at the time of PGF2α of Ovsynch was greater (P = 0.002) for control cows compared with treated cows (5.75 vs. 4.64 ng/mL). In summary, administering both PGF2α and GnRH on the same day, 7 days before the start of Ovsynch, appears to be a simple alternative that results in acceptable P/AI but potentially less progesterone during the growth of the ovulatory follicle.

Trehalose improves semen antioxidant enzymes activity, post-thaw quality, and fertility in Nili Ravi buffaloes (Bubalus bubalis).

Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profile during cryopreservation (after dilution, before freezing, and after thawing), (2) in vitro quality (after thawing), and (3) in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Semen samples (n = 20) from four buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of trehalose (0.0, 15, 30, 45, and 60 mM) and frozen in French straws. At post dilution, profile of sperm catalase (U/mL) was higher (P < 0.05) in extenders containing 15, 30, and 45 mM of trehalose as compared to control. Although profiles of superoxide dismutase (U/mL) and total glutathione (µM) were higher (P < 0.05) in extenders containing 15 and 30 mM of trehalose as compared to control. At prefreezing, sperm catalase, superoxide dismutase, and total glutathione profiles were higher (P < 0.05) in all the treatment groups as compared to control. At post thawing, the profiles of catalase and total glutathione were higher (P < 0.05) in extender containing 30-mM trehalose as compared to other treatment groups and control. Whereas, profile of superoxide dismutase was higher (P < 0.05) in extenders containing 30, 45, and 60 mM of trehalose as compared to control and 15mM group. Post thaw total sperm motility (%) was higher (P < 0.05) in extender containing 30-mM trehalose as compared to control and 15 and 60-mM groups. Although sperm progressive motility (%), rapid velocity (%), average path velocity (µm/s), straight line velocity (µm/s), curvilinear velocity (µm/s), plasma membrane (structural and functional, %), acrosome (%), and DNA (%) integrity were higher (P < 0.05) in extender containing 30 mM trehalose as compared to other treatment groups and control. The fertility rates (61% vs. 43%) were higher (P < 0.05) in buffaloes inseminated with semen doses cryopreserved in extender containing 30 mM of trehalose than the control. It is concluded that addition of 30-mM trehalose in extender improves the semen antioxidant enzymes activity, post thaw quality, and fertility in Nili Ravi buffaloes.

Effect of oestradiol benzoate on oestrus intensity and pregnancy rate in CIDR treated anoestrus nulliparous and multiparous buffalo.

The objective of the present study was to examine if administration of oestradiol benzoate (OEB) after removal of control internal drug releasing device (CIDR) could enhance oestrous intensity and pregnancy rate in anovular nulliparous and multiparous Nili-Ravi buffalo. For this, a total of 298 nulliparous (n=91) and multiparous (n=207) buffaloes received a CIDR on Day 0, and were administered PGF2α on Day 6 followed by removal of CIDR on Day 7. At Day 8, OEB was administered in approximately half of nulliparous (n=45/91) and multiparous (n=100/207) buffalo. All animals were fixed time inseminated 48 and 60h after CIDR removal, respectively. The results showed that administration of OEB but not the parity, improved oestrous intensity (3.15±0.05 vs 2.99±0.05; P=0.0026) compared to those not received OEB, respectively. However, OEB did not affect (46.2 vs 44.1; P=0.8) pregnancy per AI (P/AI). In addition, P/AI was greater (50.7 vs 39.6; P=0.036) in multiparous compared to nulliparous buffalo, respectively. The oestrous intensity (P=0.025) and response to OEB (P=0.0002) was greater in buffalo having a greater body condition (>3.0). Though, non significant, timing of ovulation was more synchronous (62.9±1.8 vs 72.4±3.6h; P>0.05) and ovulation rate was greater (91% vs 64%; P>0.05) in buffalo after OEB administration. It is concluded that administration of OEB in conjunction with the CIDR improves oestrous intensity without affecting P/AI in nulliparous and multiparous anovular buffalo.

Relationship of age to body weight, scrotal circumference, testicular ultrasonograms, and semen quality in Sahiwal bulls.

The objective of the present study was to determine the relationship of age to body weight (BW), scrotal circumference (SC), number of pixels of testicular ultrasonograms (NP), and semen quality in Sahiwal bulls. The study was based on 128 Sahiwal bulls of different age groups (from 0 to >100 months of age). Bulls were evaluated for SC, BW, and NP. Semen was evaluated once a week for five consecutive weeks from regularly collected donor bulls (n = 86) ranging in age from 25-30 to >100 months. Ejaculate volume, sperm motility, sperm concentration, sperm morphology, percent live sperms, sperm plasma membrane integrity, and normal acrosome were compared among different age groups. Mean SC and BW increased (P < 0.05) in a curvilinear manner from birth to >100 months of age. Mean NP of testicular ultrasonograms increased (P < 0.05) from 0 to 24 months and then plateaued until >100 months of age. Body weight, SC, and NP were positively correlated with age from birth until >100 months (r = 0.91, 0.87, and 0.40, respectively). Ejaculate volume (5.7 ± 0.2 vs. 4.6 ± 0.09 ml) and sperm concentration (1,281.6 ± 17.7 vs. 1,115.8 ± 55.9 × 10(6)/ml) increased (P < 0.05) in mature bulls compared to younger ones. However, motility (68.6 ± 0.3%), plasma membrane integrity (50.8 ± 1.0%), and normal acrosome (74.8 ± 0.5%) remained insignificant due to age. In six of eight age groups studied, morphological abnormalities were well within the range (18.1 ± 0.3%). In conclusion, the BW, SC, and NP of testicular ultrasonograms, ejaculate volume, and concentration increased with age. Moreover, semen quality is fairly independent of age except volume and concentration in Sahiwal bulls.

Effect of oestrus synchronization methods on oestrus behaviour, timing of ovulation and pregnancy rate during the breeding and low breeding seasons in Nili-Ravi buffaloes.

The objective of this study was to determine the effect of oestrous synchronization methods on oestrous behaviour, timing of ovulation and pregnancy rate during the breeding and low breeding seasons in Nili-Ravi buffaloes. In Experiment 1, oestrous behaviour and timing of ovulation were determined from (n=34) oestruses. The mean (+/- S.E.M.) time of ovulation after the onset of standing oestrus was greater (P<0.05) in PGF(2alpha)-induced luteolysis (30.6+/-1.5h) compared to Ovsynch buffaloes (15.0+/-0.8h). In Experiment 2, pregnancy rates were compared between two methods of synchronization (detected oestrus and Ovsynch protocol) during the breeding and low breeding seasons. Pregnancy rates of buffaloes bred at detected oestrus (62.5%) or by the Ovsynch protocol (36.3%) during the breeding season did not differ significantly (P>0.05) from those which were inseminated during the low breeding season (55.5%) and (30.4%), respectively. This study demonstrates clearly that (1) timing of ovulation in Nili-Ravi buffalo is about 30h after the onset of standing oestrus and (2) buffaloes can be successfully synchronized with optimum fertility using either PGF(2alpha) alone (detected oestrus) or using (Ovsynch protocol) during low breeding season, to calve during the period when milk availability is short.