Effects of preovulatory follicle size on estradiol concentrations, corpus luteum diameter, progesterone concentrations and subsequent pregnancy rate in buffalo cows (Bubalus bubalis).

The present study was undertaken to investigate the effects of preovulatory follicle (POF) size on estradiol concentrations, luteal profile (CL diameter and progesterone concentration) and subsequent pregnancy rate in Murrah buffalo cows. The buffalo cows (n = 49) were synchronized for estrus by two doses of PGF2α given 11 days apart. The buffalo cows were inseminated during standing estrus and again after 24 h. Ovaries were scanned at estrus and 24 h intervals until ovulation, thereafter on days 5, 12 and 16 post-ovulation to examine the POF and CL diameter. Size of POF at estrus was divided into three categories; I: 10 to ≤12; II: >12.0 to ≤14.0; III: >14.0-16.0 mm. Blood samples were collected for estradiol (on day of estrus) and progesterone concentration (on days 5, 12 and 16). The estradiol concentrations were greater (P < 0.05) in category II than category I with the greatest (P < 0.05) concentrations estimated in category III. A positive correlation (P < 0.05) between POF and progesterone concentration, CL diameter and progesterone concentration was observed on all sampling day. Pregnant buffalo cows exhibited greater (P < 0.05) plasma progesterone as compared with their non-pregnant counterpart. Greater pregnancy rates were observed with an increased size of POF (χ2 = 2.9, P > 0.05). It was concluded that the POFs having diameters between 12 and 16 mm are mature enough to be transformed into CL of such optimum diameter and can secrete optimum progesterone concentrations that can sustain the pregnancy in Murrah buffalo cows.

Impact of Buserelin Acetate or hCG Administration on the Day of First Artificial Insemination on Subsequent Luteal Profile and Conception Rate in Murrah Buffalo (Bubalus bubalis).

This study was designed to investigate the impact of buserelin acetate (BA) or human chorionic gonadotropin (hCG) administration on the day of first artificial insemination (AI) on subsequent luteal profile (diameter of corpus luteum (CL) and plasma progesterone) and conception rate in Murrah buffalo. The present experiment was carried out at two locations in 117 buffalo that were oestrus-synchronized using cloprostenol (500 µg) administered (i.m.) 11 days apart followed by AI during standing oestrus. Based on treatment (i.m.) at the time of AI, buffalo were randomly categorized (n = 39 in each group) into control (isotonic saline solution, 5 ml), dAI-BA (buserelin acetate, 20 µg) and dAI-hCG (hCG, 3000 IU) group. Out of these, 14 buffalo of each group were subjected to ovarian ultrasonography on the day of oestrus to monitor the preovulatory follicle and on days 5, 12, 16 and 21 post-ovulation to monitor CL diameter. On the day of each sonography, jugular vein blood samples were collected for the estimation of progesterone concentrations. All the buffalo (n = 117) were confirmed for pregnancy on day 40 post-ovulation. The conception rate was better (p < 0.05) in dAI-BA (51.3%) and dAI-hCG (66.7%) groups as compared to their control counterparts (30.8%). Furthermore, the buffalo of dAI-hCG group had improved (p < 0.05) luteal profile, whereas the buffalo of dAI-BA group failed (p > 0.05) to exhibit stimulatory impact of treatment on luteal profile when compared to control group. In brief, buserelin acetate or hCG treatment on the day of first AI leads to an increase in conception rate; however, an appreciable impact on post-ovulation luteal profile was observed only in hCG-treated Murrah buffalo.

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.

Comparative efficacy of E-17ß and GnRH administration on day 0 of a controlled internal drug release (CIDR) based protocol on synchrony of wave emergence, ovulation and conception rates in Murrah buffalos (Bubalus bubalis).

The study was undertaken to compare the efficacy of an estradiol-17ß + CIDR based protocol with the conventional ovsynch + CIDR based protocol for synchrony of wave emergence and ovulation in Murrah buffalos. In group I (n=25), on day 0 (beginning of experiment), buffaloes were administered a CIDR device (1.38 g P4) and concurrently received 1.5 mg E-17ß. On day 9, the CIDR was removed and a prostaglandin (PG) F2α analogue (500 µg) was administered. On day 11, buffaloes were administered a gonadotropin releasing hormone (GnRH) analogue (20 µg) and inseminated twice at 12 h and 24 h following GnRH injections. Group II (n=25) protocol was based on an ovsynch regimen plus CIDR for 7 days followed by double insemination at induced estrus. Group III (n=10) served as control and was not given any hormone on day 0 of the protocol. In groups I, II and III, the duration of new follicular wave emergences were observed on days 4.22 ± 0.12, 3.12 ± 0.33 and 5.14 ± 0.42, respectively. In group I, synchrony of wave emergence was more and the diameter of pre-ovulatory follicles was larger (P<0.05) compared to groups II and III. The first service conception rate (FSCR) was higher (P<0.05) in group I while ovulation rates were not different between groups I and II. In conclusion, more synchrony of wave emergence, larger diameter of dominant follicles and higher first service conception rate was observed following the E-17ß + CIDR based protocol in buffalos.

Comparison of follicular dynamics, superovulatory response, and embryo recovery between estradiol based and conventional superstimulation protocol in buffaloes (Bubalus bubalis).

AIM: To evaluate the follicular dynamics, superovulatory response, and embryo recovery following superstimulatory treatment initiated at estradiol-17ß induced follicular wave emergence and its comparison with conventional superstimulatory protocol in buffaloes. MATERIALS AND METHODS: Six normal cycling pluriparous buffaloes, lactating, 90-180 days post-partum, and weighing between 500 and 660 kg were superstimulated twice with a withdrawal period of 35 days in between two treatments. In superstimulation protocol-1 (estradiol group) buffaloes were administered estradiol-17ß (2 mg, i.m.) and eazibreed controlled internal drug release (CIDR) was inserted intravaginally (day=0) at the random stage of the estrous cycle. On the day 4, buffaloes were superstimulated using follicle stimulating hormone (FSH) 400 mg, divided into 10 tapering doses given at 12 hourly intervals. Prostaglandin F2α analogs (PGF2α) was administered at day 7.5 and day 8, and CIDR was removed with the second PGF2α injection. In superstimulation protocol - 2 (conventional group) buffaloes were superstimulated on the 10(th) day of the estrous cycle with same FSH dose regimen and similar timings for PGF2α injections. In both groups, half of the buffaloes were treated with luteinizing hormone (LH) 25 mg and other half with 100 ug buserelin; gonadotrophin releasing hormone (GnRH) analog at 12 h after the end of FSH treatment. All buffaloes in both protocols were inseminated twice at 12 and 24 h of LH/GnRH treatment. Daily ultrasonography was performed to record the size and number of follicles and superovulatory response. RESULTS: Significantly higher number of small follicles (<8 mm) was present at the time of initiation of superstimulatory treatment in the estradiol group compared to the conventional group (12.5±0.80 vs. 7.3±1.21, respectively, p=0.019), however, the number of ovulatory size follicles (≥8 mm) did not differ significantly between the respective groups (15.5±1.24 vs. 12.2±1.30; p=0.054). Total embryos and transferable embryos recovered were non-significantly higher in the estradiol group compared to the conventional group (5.83±0.86 vs. 4.67±1.16, p=0.328, and 3.67±0.93 vs. 2.67±0.68, p=0.437, respectively). The significant higher proportion of transferable embryos were recovered in buffaloes treated with LH compared to GnRH (73.3% vs. 48.5%; p=0.044). CONCLUSION: The average number of ovulatory size follicles (>8 mm), corpora lutea, and transferable embryos was higher in buffaloes superstimulated at estradiol-induced follicular wave compared to the conventional protocol: Further the percentage of transferable embryos was significantly higher in buffaloes administered with LH compared to GnRH.

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 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.

l-Ascorbic acid (vitamin C) supplementation to optimize health and reproduction in cattle.

Cattle can synthesize L-ascorbic acid (or Vitamin C) from either D-glucose or D-galactose through glucuronic acid pathway in the liver. L-Ascorbic acid present in cattle diet is almost totally destroyed by rumen microorganisms making them essentially dependent on its endogenous synthesis, which is assumed sufficient to meet the physiological requirement. Therefore, the role of vitamin C in cattle health and disease has remained widely overlooked. However, there is mounting evidence that the level of L-ascorbic acid in blood and other tissues decreases in association with stress and disease, and Vitamin C supplementation revealed favorable response as evident from early recovery. The present review is an attempt to summarize the existing literature pertaining to the physiological role of L-ascorbic acid and the scope of its supplementation in the prevention and treatment of diseases in cattle. It should be realized that the aqueous solution of vitamin C is highly acidic and subcutaneous or intramuscular administration may cause tissue irritation and inflammation, whereas the sodium ascorbate solution is less acidic and might be used for intramuscular administration.

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.

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.

Factors affecting the incidence of postpartum oestrus, ovarian activity and reproductive performance in Thoroughbred mares bred at foal heat under Indian subtropical conditions.

Decreased reproductive performance due to summer stress is a well known phenomenon in farm livestock. Whether this occurs in the mare and specifically how this might affect postpartum reproductive activity and performance, especially at Foal Heat (FH), is unknown. This study, therefore, aims to investigate this and the factors that might affect postpartum reproductive activity. Reproductive records of 228 Thoroughbred mares (694 mare years) bred in subtropical north-western India were retrospectively analysed. Overt oestrous activity occurred within 21 d postpartum in 92.94% (645/694) of mares. Significantly (p<0.001) more April foaling mares (97.37%, 185/190) expressed postpartum oestrous activity than those foaling in January (83.61%; 51/61) and February (88.49; 123/139). Similarly significantly (p<0.01) fewer multiparous mares failed to demonstrate oestrous activity than primiparous mares (6.12% vs.15.07%; 38/621 vs. 11/73, respectively). 190 of these 694 mares were additionally monitored to confirm ovulation; in these mares onset of FH (oestrus plus confirmed ovulation) occurred 8.42+/-0.17 d and first ovulation 13.64+/-0.20 d postpartum. Month, stud farm, year, and parity did not affect interval from parturition to FH onset or to first ovulation; or FH onset to ovulation. In FH bred mares Day 16 pregnancy rate and overall foaling rate were 53.76% (100/186) and 46.24% (86/186) respectively and were similar to those of mares bred later postpartum. FH pregnancy rates were not affected by stud, season, month, year, number of matings, or day of ovulation but were significantly (p<0.008) lowered by increasing mare age. Significantly (p<0.01) lower Day 16 pregnancy rates were observed in uterine treated mares compared to untreated mares (31.09% vs. 57.96%; 9/29 vs. 91/157, respectively), this difference was not evident during the rest of pregnancy. In conclusion, postpartum reproductive and ovarian activity appears to be affected by environment, i.e., delayed in subtropical kept Thoroughbred mares compared to those kept in temperate climates. However, resulting reproductive performance at FH and the factors affecting postpartum reproductive activity are similar.

The pattern of embryonic fixation and its relationship to pregnancy loss in thoroughbred mares.

Ultrasonographic pregnancy records of 195 mares from six Thoroughbred stud farms, over a period of 7 years were retrospectively analysed to assess the effect of various factors on embryonic vesicle (EV) fixation pattern and pregnancy outcome. Of the total of 746 pregnancies analysed, significantly (p < 0.01) more EV fixations were evident in the right uterine horn than in the left (53.35% vs 46.65% respectively). There was no significant effect of either, the side of ovulation, or age of the mare, on the side of EV fixation. However, EV fixation, was significantly (p < 0.001) more likely to occur in the right uterine horn in maiden and barren mares (65.75% vs 57.45% respectively). The age and reproductive status of the mare as well as foal heat breeding failed to demonstrate a consistent effect on pregnancy loss relative to the side of EV fixation. In lactating and foal heat bred mares, EVs were significantly (p < 0.0001) more frequently established in the contralateral horn to the one from which the mare delivered her most recent foal. In lactating mares, significantly (p < 0.05) higher embryonic and pregnancy losses were observed in the ipsilateral horn. In conclusion, (a) side of EV fixation was (i) independent of the side of ovulation and mare age (ii) significantly (p < 0.001) affected by reproductive status, (b) neither age of mare nor reproductive status had any effect on pregnancy loss rates relative to the side of EV fixation and (c) in lactating mares the EV had a greater chance of fixation and survival in the horn contralateral to the one from which the mare delivered her most recent foal.

Reproductive efficiency of Thoroughbred mares under Indian subtropical conditions: A retrospective survey over 7 years.

Service records of 253 mares (1181 mare-years) spanning over 7 consecutive years, from nine organized Thoroughbred stud farms, situated in the subtropical northwestern India were retrospectively analyzed to assess their reproductive performance. The overall per cycle pregnancy rate at Day 16 and overall foaling rates were 50.30% and 68.95%, respectively, and were significantly higher in mares aged 3-7 years than > or =18 years old mares. The late embryonic losses (9.86%) that occurred between Days 16 and 39 post-ovulation contributed more than 50% of the overall detected pregnancy losses (19.11%). The overall percent detected pregnancy losses were lower in mares at ages 3-7 years compared to those at ages > or =18 years (14.78% vs. 46.43%, respectively; P<0.0001). Chronic barren and habitual aborter mares tended to affect reproductive efficiency of mares. Fifty percent of the mares that experienced > or =2 consecutive abortions or barren years, again stayed aborted or barren in the next seasons, respectively. No effect of numbers of matings per oestrus was observed on overall fertility. Neither the induction of oestrus nor ovulation by exogenous hormonal treatment had any effect on most of the analyzed reproductive parameters. Regarding breeding month or years, the reproductive efficiency did not differ significantly. The incidence of multiple pregnancies was 5.40% and percent late embryonic loses were higher (P=0.0016) in twin (21.98%) than singleton (8.64%) pregnancies. In conclusion, comparatively lower fertility rates were recorded in Thoroughbred mares bred under Indian subtropical climatic conditions than those reported from temperate regions that might be due to difference in breeding management rather than prevailing environment.

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 (

Estrus induction and fertility rates in response to exogenous hormonal administration in postpartum anestrous and subestrus bovines and buffaloes.

A total of 130 animals (82 cattle, 48 buffaloes) with histories of anestrous 60-90 days post-partum and belonging to different agroclimatic zones of Punjab were subjected to rectal palpation and blood samplings at least three times at weekly intervals. The body condition score (BCS) of each animal was also recorded. The animals were divided into two groups; viz., true anestrous (Gp-I) and subestrus (Gp-II) through rectal palpation of ovaries and plasma progesterone (P4) concentrations. Furthermore, the Gp I and II animals were divided into treatment (Gp Ia, 40 cattle and 16 buffaloes; Gp IIa, 12 cattle and 14 buffaloes) and control groups (Gp Ib, 20 cattle and 8 buffaloes; Gp IIb, 10 cattle and 10 buffaloes). True anestrous animals (Gp Ia) were treated with 3 injections of hydroxyprogesterone caproate (750 mg, i.m.) at 72-hr intervals followed by injection of equine chorionic gonadotropin (eCG; 750 I.U., i.m.) 72 hr after the last progesterone injection. The animals were bred at the first estrus after the induced one. The first service conception rate (FSCR), overall conception rate (OCR), services per conception and pregnancy rate of the true anestrous treated cattle (Gp Ia) were 44.4%, 48.0%, 2.08 and 60.0%, respectively. In the true anestrous control cattle (Gp Ib), only five that were observed to be in estrus failed to conceive. In the anestrous treated buffaloes (Gp Ia), the FSCR, OCR, services per conception and pregnancy rate were 50.0%, 62.5%, 1.6 and 62.5%, respectively. No buffalo amongst true anestrous control (Gp Ib) showed estrus. The subestrus animals (Gp IIa) were administered Prostaglandin F(2alpha) (PGF(2alpha); 25 mg Dinoprost, i.m.) and bred at induced estrus. Amongst the Gp IIa animals, all cattle (100%) and twelve buffaloes (85.7%) responded to treatment. Of these animals, the FSCR and pregnancy rate at induced estrus in the cattle were 50.0% each, whereas they were 66.6% and 57.1%, respectively, in the buffaloes. The subestrus control animals (Gp IIb) remained infertile. In summary, the plasma P(4) profile can be used to differentiate true anestrous and subestrus animals and thus to determine a hormonal therapy. Furthermore, fertile estrus can be induced with hormonal therapy in anestrous and subestrus bovines.

Prevalence of aerobic and anaerobic uterine bacteria during peripartum period in normal and dystocia-affected buffaloes.

Parturition complications predispose establishment of uterine infections, which in turn affect subsequent fertility. The aim of present study was to characterize and compare the type of bacterial flora prevalent within the uterine lumen of dystocia-affected buffaloes and compare them with the normally calving buffaloes. The study was conducted on 40 buffaloes; of which 10 calved normally (Group I) and 30 were treated for dystocia (Group II). Bacteriological examination was performed using uterine swabs, which were collected before delivery, immediately after delivery and day's 24-60 postpartum. A total of 30 uterine swabs from Group I and 79 swabs from Group II were collected, of which 19 (63.3%) and 71 (89.9%) yielded significant bacterial growth, respectively. A total of 205 isolates belonging to 10 different genera of bacteria were identified, 8 facultative anaerobes and 2 obligate anaerobes. In Group II, 91.6% of the bacteria positive swabs (n = 71) yielded mixed cultures, whereas the remainder being pure cultures. In contrast, 89.5% of the bacteria positive swabs of Group I (n = 19) yielded pure cultures. Mixed infections comprised mostly Arcanobacter (Actinomyces) pyogenes together with obligate anaerobes, Fusobacterium spp. and Bacteroides spp. In Group II, the frequency of incidental and coliform group bacteria was highest at the time of parturition, i.e., before and immediately after delivery, and decreased to nil during the 24-60-day postpartum period. However, in Group I, the incidental and coliform group of bacteria present at the time of parturition apparently persisted beyond the period when uterine involution is complete. The frequency of obligate anaerobes and A. pyogenes at the time of parturition was nil in the Group I while they predominated in dystocia-affected buffaloes (Group II). During the postpartum period of 24-60 days, the frequency of both obligate anaerobes and A. pyogenes increased significantly in Group II, whereas in Group I, only occasional isolates were obtained. To conclude, at the time of calving the prevalence of obligate anaerobes and A. pyogenes occurring in combination was highest in dystocia-affected buffaloes, and further increased in the postpartum period suggesting that these infections act synergistically.

Quantitative unbiased estimates of endometrial gland surface area and volume in cycling cows and heifers.

Entire reproductive tracts were removed from seven normal healthy heifers and qualitative unbiased estimates made of endometrial gland volume density and glandular surface density. After examining approximately 55 microscopic fields of endometrium from each tract, a mean glandular surface density value of 10.2 mm(2)/mm(3) (CE 3.1%) was obtained. The stereological method was then employed in optimising the design of the main study. The endometrial height was measured for 17 healthy cycling heifers and 19 similar cows. Subsequently, unbiased estimates were made of intercaruncular endometrial gland volume per unit surface for all cattle were investigated; differences between heifers and cows generally, and the possible effect of the follicular and luteal phases of the oestrous cycle were compared. The mean surface area of glands per unit area of endometrium at the intercaruncular site in heifers and cows was approximately 18 mm(2)/mm(2) in the follicular phase and 26 mm(2)/mm(2) in the luteal phase, figures similar to the gland area found in women. The intercaruncular gland volume increased significantly, by about 30% during the luteal phase of the bovine oestrous cycle in heifers, from 0.01 to 0.13 per mm(3). The differences in endometrial anatomy between site of sampling and either follicular or luteal phases of the oestrous cycle were always more significant in heifers than cows. The endometrial thickness in cows was always greater than for heifers, irrespective of the site of sampling. It was concluded that the intercaruncular endometrium of cattle was far more active physiologically than recognised previously.

Some aspects of immunology of the bovine uterus related to treatments for endometritis.

Endometritis in breeding cattle occurs during the postpartum period, and is associated primarily with contamination of the reproductive tract involving Arcanobacter pyogenes (formerly Actinomyces pyogenes) together with Gram-negative anaerobes. Polymorphonuclear inflammatory cells (PMNs) contribute partly to the defense mechanisms against micro-organisms contaminating the vagina and uterine lumen, whose phagocytic activity depends on bacterial opsonisation by humoral antibodies; significant numbers of lymphocytes are also present. Whilst leukocyte numbers in the uterine lumen are relatively high during metoestrus and dioestrus compared to other phases of the oestrous cycle, their functional activity is unaffected. Humoral antibody concentrations in the reproductive tract are stimulated following exposure to local antigen, and the response is site dependent; of the several different classes of immunoglobulins, IgG predominates in the uterus and IgA the vagina. Only a portion of the total IgG1 found on the uterine lumen is synthesised locally in the endometrium, the remainder and all of the IgG2 is derived from the local uterine blood supply. Generally, concentrations of immunosuppressant proteins present in the uterine lumen increase under progesterone dominance, and these inhibit lymphocyte proliferation, making the uterus more susceptible to infection. The relationship between uterine susceptibility to micro-organism contamination and the luteal phase of the oestrous cycle is still unclear. Intrauterine infusion of immunomodulators such as E. coli lipopolysaccharides (LPS) or oyster glycogen, in healthy cows and those with endometritis, stimulates leukocytes to migrate into the uterine lumen. At a dosage rate of 100 microg, lipopolysaccharides are not absorbed by the healthy endometrium and do not alter the oestrous cycle length. It is unknown, whether a similar dose can be absorbed through an inflamed endometrium in naturally occurring cases of endometritis to cause systemic illness. Currently, prostaglandin F2alpha is recommended for treating endometritis in both cycling and non-cycling cows, but its mode of action in non-cycling cows is not fully understood. The efficacy of endometritis treatment using an intrauterine infusion of an immunomodulator in cases occurring naturally has not been determined on a large scale.

Effectiveness of lipopolysaccharide as an intrauterine immunomodulator in curing bacterial endometritis in repeat breeding cross-bred cows.

Antibiotics are usually used to combat microbial infections of the uterus, responsible for hindering establishment of pregnancy in cross-bred cows. The major disadvantages of antibiotics are: development of bacterial resistance, high costs and diminishing uterine defense mechanisms (UDM). As an alternative therapy, intrauterine application of Escherichia coli Lipopolysaccharide (E. coli LPS) as a uterine defense stimulator was used in this study in confirmed clinical cases of repeat breeding associated with bacterial endometritis. In the treated group (n=12), on the day of estrus, 100 microg of E. coli LPS dissolved in 30-ml sterile phosphate buffer saline (PBS) was infused intrauterine; while in the control group (n=12), only 30 ml of PBS was infused. Six-hour post-treatment, in the treatment group uterine washings showed a 100-fold increase in the total leucocytic count (TLC). Out of the cellular contents, more than 80% of the cells were recognised as neutrophils; above 60% were alive and their phagocytic activity was five bacteria/neutrophil. Such a cellular response was maintained until 72-h post-treatment. At the subsequent estrus period, the cervicovaginal mucus (CVM) became clear in 9 out of 12 cows (75%) and showed no bacterial growth. In the control group, similar micro-organisms were present in CVM of all the 12 cows before and after the PBS infusions. During the subsequent estrus, all nine cows with sterile CVM in the treatment group conceived while only one cow conceived from the control group. It was concluded that, administration of intrauterine E. coli LPS as single infusion in cows with bacterial endometritis stimulated UDM and cleared the infection within one estrous cycle, and thereby restoring fertility.