Experimental treatment of summer anoestrous — Buffaloes with norgestomet and prid

Two experiments were undertaken to treat summer anoestrous buffaloes drawn from two distinct levels of total management, i.e., village vs. farm. At both locations, animals were randomly divided into three groups. One group was treated with norgestomet implants^a plus 700 I.U. PMSG at implant removal; the second group was treated with progesterone releasing intravaginal devices (PRID)^b and the third group was retained as untreated control. Serum samples were collected immediately before the commencement of treatments, before the withdrawal of treatments and on day 10 post-insemination (Artificial Insemination done 48 and 72 hours after the withdrawal of treatments). The samples were analysed for progesterone by radioimmunoassay. The ovulation and conception rates of norgestomet-treated animals were 58.33% vs. 77.79% and 40.16% vs. 60.11%, respectively, at village vs. farm. Similarly, the ovulation and conception rates of PRID-treated animals were 33.33% vs. 38.33% and 8.33% vs. 27.77%, respectively, at village vs. farm. None of the control animals exhibited oestrus during the course of the study.     

Pregnancy in suboestrus buffaloes (Bubalus bubalis) after treatment with Prostaglandin F(2) alpha

Seventy six apparently anoestrous buffaloes, with a palpable corpus luteum at day 43 to 547 post partum, were treated intramuscularly with Prostaglandin, using either 25 mg single dose (Group I), 25 mg double dose 11 days apart (Group II), or 50 mg single dose (Group III). Animals exhibiting oestrus, without any treatment, during the period of experimentation served as control (Group IV). The overall conception rate for the treated animals was 28.8, 51,5, and 69.7 percents after one, two, and three inseminations, respectively. The conception rate, at induced as well as subsequent oestrus, was comparable to control animals. The intergroup differences among treated animals were not significant. The conception rate was greater when the induced heat was more intensely expressed. The subsequent mean oestrus cycle length was similar to controls when all of the treated animals were considered together.     

Follicular dynamics and temporal relationships among body temperature, oestrus, the surge of luteinizing hormone and ovulation in Holstein heifers treated with norgestomet

The effects of chronic treatment with norgestomet on follicular dynamics, corpus luteum growth and function as well as the temporal relationships among body temperature, oestrous behaviour, the luteinizing hormone (LH) surge and ovulation following implant removal were studied in 16 Holstein heifers. Oestrous cycles of the heifers were initially synchronized using 2 injections of prostaglandin F-2 alpha (PGF-2 alpha) 12 days apart. The heifers were then implanted with a norgestomet ear implant for 9 days, beginning either at the middle of the synchronized cycle (dioestrus) or at the end of the synchronized cycle (pro-oestrus). Follicular dynamics, corpus luteum growth and regression, and plasma progesterone were not affected by norgestomet treatment at dioestrus. The dominant follicle present at the time of norgestomet implantation in the pro-oestrus group was maintained during the 9-day implant period of 6 of 8 heifers and ovulated after implant removal. Time from implant removal to onset of standing oestrus and time to LH peak following implant removal were highly correlated with the time of ovulation (r = 0.92 and 0.96, respectively). Onset of standing oestrus and the LH peak and the onset of standing oestrus and peak vaginal and rectal temperatures were also highly correlated (r = 0.96, 0.82 and 0.81, respectively). It is concluded that any decrease in pregnancy rates following treatment with norgestomet is not due to asynchrony among oestrus, the LH surge and ovulation.     

Induction of ovulatory estrus and fertility in non-cycling buffaloes with norgestomet during low breeding season

Field trials were designed to evaluate use of norgestomet treatment to induce ovulatory estrus in non-cycling buffalo cows and heifers during low breeding season. Twenty-five buffalo cows and 50 heifers under village management were given a 9-day treatment with a polymer implant containing 6 mg norgestomet with IM injections of 5 mg estradiol valerate + 3 mg norgestomet at the time of implantation and 600 IU PMSG when the implant was removed. Fifty animals served as controls without any treatment. Seventy-four treated animals showed estrus during the period between 36 to 80 hours after removal of the implant. Twenty-five buffalo cows and 40 heifers that could be further followed up were inseminated twice at 8-hour intervals, 12 hrs after induction of estrus with chilled semen by recto-vaginal method. Of these, 15 (23.1%) conceived, 9 (36%) among buffalo cows and 6 (15%) among heifers. Fourteen buffalo cows and 30 heifers that did not conceive manifested cyclic estrus at an interval of 22.4 and 20.6 days. The conception rate in the cyclic estrus was 57% and 23.3%, respectively, for buffalo cows and heifers. The overall conception rate over two inseminations was 46.2%, 68% in buffalo cows and 32.5% in heifers. In the control group, five (10%) showed spontaneous estrus and two (40%) conceived during the period of the experiment.     

Effects of repetitive norgestomet treatments on pregnancy rates in cyclic and anestrous beef heifers

Sixty-eight 12- to 14-month-old crossbred beef heifers averaging 285 kg were assigned at random to treated (n = 35) and control (n = 33) groups to evaluate the use of repetitive norgestomet treatments. Treated heifers received an ear implant containing 6 mg norgestomet on two occasions 16 days apart. Injections of 3 mg norgestomet and 5 mg estradiol valerate (EV) were given intramuscularly the same day as first implantation (Syncro-Mate-B). Implants were removed after eight days. Four bulls were then placed in each of two pastures containing half of the treated and half of the control heifers for 24 days after the time of the first implant removal. Progesterone concentrations from blood samples collected prior to the first treatment were used to determine reproductive status. The overall pregnancy rate 64 days after first implant removal for treated anestrous heifers (61%; 14 23 ) was similar (p > 0.25) to untreated (73%; 11 15 ) and treated (75%; 9 12 ) cyclic heifers, but higher (p < 0.1) than for untreated anestrous heifers (33%; 6 18 ). This treatment advantage resulted from an increased (p < 0.01) pregnancy rate after the second implant removal. In summary, repetitive norgestomet treatments enhanced pregnancy rate in anestrous heifers within a 24-day breeding season.     

Reproductive hormone secretions and first service conception rate subsequent to ovulation control with Synchro-Mate B

Two groups of beef females receiving suboptimal energy diets were treated with Synchro-Mate B to control ovulation. The first group consisted of 30 suckled cows and 16 heifers. These females were bled 10 days and immediately prior to the implantation of norgestomet implants, at implant removal, 24, 27, 30, 33 and 36 hours and 9 and 16 days post-implant removal. The second group which consisted of 40 cows and 8 heifers was handled in the same manner except no blood samples were collected from 24 to 36 hours following implant removal. Calves were removed from all the cows for 48 hours, beginning at implant removal. All animals were artifically inseminated 48 hours following implant removal. Blood plasma was assayed for concentrations of progesterone and LH. The first service conception rate was 21% and 40% for groups 1 and 2. Several factors were identified that reduced the first service conception rate. In summary, Snychro-Mate B is an effective method to synchronize estrus in cattle. However, stress subsequent to implant removal should be avoided in order to obtain a higher first service conception rate.     

Effect of norgestomet on peripheral levels of progesterone and estradiol-17beta in beef cows

Peripheral levels of progesterone and estradiol 17beta were quantified in 27 cycling cows following administration of a single Hydron ear implant (G. D. Searle and Co.) containing 2, 4 or 6 mg norgestomet or controls which received no implant. Implants were inserted subcutaneously in the ear on day 15 of the estrous cycle (day of estrus = day 0) and removed 9 days later. The 4 mg (seven of seven cows) and 6 mg (six of six cows) implants suppressed estrus; however, three of eight cows in the 2 mg group exhibited estrus prior to implant removal. The 6 mg implant group had a significantly longer interval from implant removal to estrus than either the 2 or 4 mg group. Failure to detect differences in the rate at which progesterone declined indicated norgestomet treatment did not affect normal corpus luteum regression. Estradiol levels rose at a similar rate approaching estrus in all treatments. There was no indication of increased endogenous estradiol levels due to norgestomet treatment.     

Effects of short-term treatment with progesterone superimposed on 11 or 17 days of norgestomet treatment on the interval to oestrus and fertility in Bos indicus heifers

The aims of this study were to determine: (1) if short-term treatment of Bos indicus heifers with progesterone (P₄) while implanted with a s.c. norgestomet implant for 17 days would influence the time interval to oestrus and increase fertility of the synchronised oestrus, and (2) whether the response to treatment with P₄ would differ between heifers treated with a norgestomet implant for 17 vs. 11 days when short-term treatment with P₄ is applied 3 days prior to implant removal. B. indicus heifers at two separate sites (A and B) were allocated to three groups at each site. Heifers in two groups (NG and NGP₄ groups) were given a single s.c. norgestomet implant on the first day of treatment (day 0) while heifers in a third group (NGP₄PG group) were implanted on day 6. A single P₄ releasing Controlled Internal Drug Release device (CIDR) was inserted on day 14 in heifers in the NGP₄ and NGP₄PG groups and was removed 23.5±0.07 h later (day 15). Heifers in the NGP₄PG group were administered an analogue of prostaglandin F_2α (PGF_2α) at the time of CIDR removal to regress corpora lutea. Implants were removed from all heifers on the same day (day 17) and a 400 IU of equine chorionic gonadotrophin (ECG) was administered s.c. Animals were artificially inseminated 11.1±0.17 h after detection of oestrus, using frozen semen from one bull at site A and one of five bulls at site B. Inseminations were carried out by one of two technicians. Treatment with P₄ delayed oestrus and reduced the synchrony of oestrus at site A (hours to oestrus±SD: NG group, 39.0±13.7; NGP₄ group, 66.3±24.4; NGP₄PG group, 58.9±20.5 h; P<0.05) but not at site B (41.4±15.2, 42.5±10.1, 45.4±10.3 h; P>0.05). Pregnancy rates 6 weeks after insemination were found to be significantly associated with bull (P<0.001), treatment group (P=0.013) and insemination technician (P=0.033). Pregnancy rates were greater in the heifers in the NGP₄ group than heifers in the NG group [50.3% (78/155) vs. 36.4% (60/165); odds ratio=1.83, 95% CI=1.14 to 2.96] and similar between heifers in the NGP₄ and NGP₄PG groups [50.3% (78/155) vs. 51.1% (63/117); odds ratio=1.06, 95% CI=0.67 to 1.69]. It was concluded that acute treatment with P₄ can improve pregnancy rates in B. indicus heifers treated for 17 days with norgestomet implants. Reducing the duration of norgestomet treatment to 11 days and administration of PGF_2α at the time of ending treatment with a CIDR device resulted in no differences in fertility, mean intervals to oestrus or synchrony of oestrus.     

Plasma progesterone profiles and fertility status of anestrus Zebu cattle treated with norgestomet-estradiol-eCG regimen

Zebu cattle are notorious for poor fertility characterized by late maturity and long intercalving intervals attributed to a variety of factors, including genetic, nutritional and climatic. The aim of the present investigation, therefore, was to induce fertile estrus in acyclic pubertal heifers and postpartum anestrous Zebu cows by hormonal intervention. Pubertal Hariana and Sahiwal anestrous heifers (n=51) and postpartum cows (n=55) were either assigned a placebo (controls, N=6 for each breed and parity) or treated with 10-d norgestomet (3 mg) subcutaneous ear implants, with an initial injection of 3 mg, im norgestomet + 5 mg estradiol valerate, followed by 500 IU eCG at implant withdrawal (NOR-treated groups). Jugular venous plasma samples were obtained from a total of 28 animals (controls : 4 heifers and 4 cows; NOR-treated : 12 heifers and 8 cows) on Days 0 (implant insertion), 3, 7, 9 and Day 10 (implant withdrawal), every 12 h on Days 11 and 12, and then once daily on Days 17, 24 and 31. All the samples were assayed for progesterone. Almost all (97%) heifers and 81% cows were induced to estrus, the majority (92% heifers and 79% cows) within 120 h of implant removal. Synchrony of the induced estrus was better in cows, but interval to estrus and estrus duration were significantly longer in heifers (P<0.05). Post-treatment fertility, based on Day 28 nonretum rate, first service, and overall conception rates, was better in heifers (78.9, 60.5 and 73.7%, respectively) than cows (77.1, 48.6 and 62.9%, respectively), but the differences were significant only for the overall pregnancy rate (71.8% for heifers and 51.2% for cows; P<0.05). Low pre-treatment plasma progesterone values (<0.5ng/mL) were consistent with ovarian inactivity, confirming the true anestrus status of experimental animals. Controls failed to exhibit estrus and maintained low progesterone concentrations throughout the study. In treated animals, high progesterone values from Day 17 onwards suggested ovulatory estrus. These early luteal phase progesterone concentrations in nonpregnant (P=0.06) and nonpregnant, nonretum (P<0.05) animals were low in comparison with those of pregnant animals. Good fertility resulting from breeding according to estrus, inspite of variable intervals to estrus and estrus duration, advocates its advantage over fixed-time insemination in norgestomet-treated anestrous Zebu cattle.     

Improving the reproductive performance of Egyptian buffalo cows by changing the management system

The objective of the present study was to determine the possibility to improve the reproductive performance of buffalo cows through the continuous exposure to bull with grazing and free-stall housing management. Sixty-four Egyptian multiparous buffalo cows raised under two different management systems in two farms were used in this study. The cows in the first farm (management system 1, MS1) were loose--housed in a free-stall yard, grazed for 4 h per day, suckled their calves for 2-3 months and were continuously exposed to a fertile bull. The cows in the second farm (management system 2, MS2 ) were confined in an open-fronted tie-stall shed, not grazed, suckled their calves for only 7 days and were exposed to a fertile bull twice per day (30 min per session). All the cows were fed a diet of green berseem (Trifolium alexandrinum), rice straw and concentrates to meet their maintenance and production requirements. The cows during both the treatments were milked twice per day after weaning. The cows in both groups were between the second and the sixth parity, weighed 450-480 kg and had average daily milk yields of 5.0-6.0 kg. In each farm, cows were visually checked twice daily at 07:00 and 17:00 h for the signs of oestrus and animals proved standing heat were naturally mated. Rectal palpation was used to monitor uterine involution and for pregnancy diagnosis. Blood was sampled twice per week from 7 to 150 days post-partum for serum progesterone assay. The results revealed that post-partum intervals to each of first ovulation, first oestrus, conception and next parturition were significantly (P < 0.05) shorter in MS1 group than in MS2 group. In the meantime, MS1 increased (P < 0.01) the conception and calving rates by 21 and 25%, respectively compared to MS2. Percentages of post-partum cyclic animals and animals exhibiting ovulatory oestrus were greater (P < 0.01) in MS1 group than in MS2 group. However, the percentage of animals cycling before day 60 post-partum was significantly (P < 0.01) lower in MS1 group than in MS2 group (13% versus 28%). By day 120 post-partum, only 63% of the buffaloes were cycling in MS2 group versus 94% in MS1 group. Percentage of silent ovulation was insignificantly higher in MS2 group (34%) than in MS1 group (25%). However, the percentage of false oestrus was higher (P < 0.01) in MS1 group than in MS2 group (16% versus 3%). In addition, percentage of short ovulatory cycles (15-17 days) was greater (P < 0.01) in MS1 group than in MS2 group, whereas percentage of long ovulatory cycle (25-28 days) was higher (P < 0.01) in MS2 group than in MS1 group. It was concluded that continuous exposure of buffalo cows to a fertile bull with grazing management under free-stall housing system enhances resumption of post-partum ovarian activity and improves conception and calving rates.     

Syncro-mate B induces estrus in ovariectomized cows and heifers

Eleven ovariectomized Hereford x Simmental cows and 10 ovariectomized crossbred heifers (primarily Angus and Hereford) were given the Syncro-Mate B (SMB) estrous synchronization treatment. The SMB treatment consisted of a 2 ml i.m. injection containing 5 mg of estradiol valerate and 3 mg of norgestomet plus a hydron ear implant containing 6 mg of norgestomet. The ear implant was removed 9 d later. Cows and heifers were considered in estrus only if they stood for mounting by a herdmate or a bull. Observations for estrus were made four or six times each day for 3 d after implant removal. The 21 animals were used in eight trials. Each trial involved 9 or 11 cows or 5 or 10 heifers. Four days to three weeks elapsed between implant removal and implant insertion for the next trial. No ovariectomized cow or heifer was observed in estrus for 21 d before treatment with SMB. In the eight trials, 3 of 9, 7 of 9 and 6 of 11 cows exhibited estrus, whereas 5 of 10, 1 of 5, 3 of 5, 3 of 5 and 5 of 5 heifers exhibited estrus after treatment. When data were pooled, 16 of 29 (55.2%) cows and 17 of 30 (56.7%) heifers exhibited estrus after treatment. Our data indicate that the SMB treatment can induce estrus in cows and heifers, independently of the ovaries.     

Synchronization of estrus and fertility in goats with norgestomet ear implants

Estrus was synchronized in 64 dairy goats in July with norgestomet ear implants. Half the does received ear implants that contained 6 mg norgestomet and the remaining does received implants that contained 3 mg. Implants were left in place for 11 days. Each doe received i.m. injections of 400 IU PMSG and 50 mug cloprostenol 24 hours prior to implant removal. Twenty-eight of 32 does (87.5%) that received 6 mg or 3 mg norgestomet exhibited onset of estrus within 24 hours of implant removal. All does had exhibited onset of standing estrus by 43 hours after implant removal. Does were hand-mated to fertile bucks twice daily while in standing estrus. There were no differences between does implanted with 6 mg or 3 mg in fertility to the induced estrus (74.2% vs 75% kidding), mean length of gestation (151.0 +/- 3.2 vs 151.6 +/- 2.0 days), mean number of kids per doe (2.1 +/- 0.8 vs 2.3 +/- 0.7) or in mean kid weights (3.10 +/- 0.80 vs 3.06 +/- 0.86 kg) (6 mg vs 3 mg, respectively). It was concluded that ear implants that contained 3 mg of norgestomet were equally as effective as implants that contained 6 mg for synchronization of estrus in dairy goats.     

Effects of estradiol-17β and hCG supplementation on superovulatory responses and embryo quality in swamp buffalo (Bubalus bubalis) implanted with norgestomet

Twenty-four cycling swamp buffaloes with normal reproductive histories and 2–3 months postpartum were used to investigate the effect of addition of estradiol-17β and human chorionic gonadotrophin (hCG) to the superovulation regime on the level of ovarian stimulation and embryo production.The estrous cycles of buffaloes were synchronized by prostaglandin injection and then divided into two groups for superovulatory treatment. Those in Group 1 (n = 12) received a implant containing 3 mg norgestomet (Syncro-Mate-B) for 9 days (insertion day is Day 0), with 4000 IU of equine chorionic gonadotrophin (eCG) and 500 μg cloprostenol i.m. given at Day 7. Group 2 (n = 12) received the same regime as Group 1, together with 7.5 mg estradiol-17β given in three intramuscular injections on Days 3, 5 and 7 in decreasing doses (4.0, 2.5 and 1.0 mg, respectively) and 5000 I.U hCG i.v. coincidentally with the first insemination. Estrus was monitored visually and by placing treated animals with bulls. Each animal was inseminated twice with frozen sperm after standing estrus. The numbers of corpora lutea (CL) and follicles greater than 8 mm in diameter were recorded via palpation per rectum at 6 days after implant removal. Two days later 11 animals from Group 2 and two from Group 1 were slaughtered for direct observation of ovarian responses and for embryo collection.The mean number of CL were 0.91 ± 0.66 and 9.08 ± 5.0 for Groups 1 and 2, respectively. The average recovery rate based on CL counts at slaughter was 60% in Group 2. No embryos were recovered from the two animals in Group 1. Seventy-nine percent of the collected ova were fertilized and more than 60% of them had developed into hatched blastocysts. The percentages of buffalo with excellent and good estrus were 41.6 and 91.6% for Groups 1 and 2, respectively.These results showed that the supplementation of estradiol-17β and the hCG treatment significantly improved the level of ovarian stimulation in swamp buffalo.     

Reproductive performance of synchronized beef cows as affected by inhibition of suckling with nose tags or temporary calf removal

A study was conducted to determine whether presence of the calf during suckling inhibition influences the response to estrus synchronization in beef cows. Angus or Hereford cows (n=89) were administered Syncro-Mate-B (SMB), which consisted of a 6-mg norgestomet ear implant (in situ 9 d) in conjunction with 5 mg of estradiol valerate and 3 mg (im) of norgestomet. Cows were allotted by breed, body condition, stage of the estrous cycle, parity and date of parturition to 1 of 3 treatments: 48-h calf removal; ad libitum suckling; or inhibition of suckling with a nose tag for 48 h. Calves were weighed at time of SMB implant removal, 48 h later and at weaning. Cows were mated via AI approximately 12 h after detection of estrus during a 30-d period after implant removal followed by a natural service period of 35 d. At 48 h after implant removal, calf removal and nose tag calves had lost an average of 3.6 and 0.9 kg, respectively, while the suckled calves gained 1.8 kg (P < 0.01). Mean calf weight at weaning did not differ among treatments. Synchronized estrous response (within 5 d of implant removal) was not different among treatments. Pregnancy rate for cows exhibiting a synchronized estrus (5 d AI) for calf removal, nose tag and suckled cows was 76, 48 and 48%, respectively (P>0.10). Treatment did not affect the 30-d AI or overall 65-d pregnancy rate. In this study, there were no differences observed in the percentage of synchronized or pregnant cows following suckling inhibition by either a nose tag or calf removal. Transient reductions of calf body weight during the 48-h calf removal period did occur in both the nose tag and calf removal groups.     

Synchronization of estrus in beef heifers with a norgestomet implant and prostaglandin F2alpha

In a 5-year study (1973-1977), 281 cycling beef heifers were treated with a 7-day norgestomet (SC21009) ear implant and an intramuscular injection of prostaglandin F(2alpha) (PGF(2alpha)) at the time of implant removal or 24 hr before implant removal. Percentages of heifers in estrus by 36, 48, 60, 72, and 120 hr after implant removal were 32.4, 52.7, 71.6, 80.1, and 93.2, respectively. Onset of estrus occurred an average of 49.8 +/- 4.7 hr after treatment. Percentages of heifers in estrus 36 hr after treatment were 5.7 and 51.7 for those with a corpus luteum and those without a corpus luteum (or determined regressing by palpation) at implant removal, respectively. When PGF(2alpha) was injected 24 hr before implant removal, 55% of the heifers were in estrus by 36 hr after implant removal compared to 30% when PGF(2alpha) was injected at the time of implant removal; however, by 60 hr after implant removal the difference was 76% vs. 71%. First-service conception rates for synchronized and nonsynchronized heifers were 62.2% and 59.6%, respectively. During 1976 and 1977 heifers were checked for estrus every 4 hr and inseminated 2, 6, 10, 14, 18, 22, 26, or 30 hr after first detected to be in standing estrus. Conception rate was not significantly affected by time of insemination but tended to be higher for heifers bred 26 and 30 hr after first being detected in standing estrus (78.9% and 70.0% vs. average 59.2%). Treatment with a 7-day norgestomet implant plus a single injection of PGF(2alpha) 24 hr before or at implant removal appears to be a practical technique for synchronizing estrus in cycling heifers without affecting conception.     

Follicle development in cyclic, anoestrous and FSH-treated brushtail possums (Trichosurus vulpecula)

The common brushtail possum (Trichosurus vulpecula) is a pest of considerable economic importance in New Zealand. Attempts to develop methods of suppressing reproduction in this species are currently hampered by the lack of reliable methods to synchronise oestrus and ovulation in this species. The objective of this study was to compare antral follicle populations in anoestrous and cyclic brushtail possums and to assess the efficacy of exogenous FSH to induce follicle development in anoestrous animals. Ovaries were recovered from anoestrous possums after administration of either exogenous FSH (1.0 mg/injection) or the saline vehicle alone (0.5 ml/injection) at 12-h intervals for 3 days (n = 6/group), and from cyclic animals (n = 6) that were euthanised in mid-follicular phase (5 days after removal of their pouch young). All antral follicles > or =1.0 mm in diameter were dissected free of extraneous tissue, incubated in vitro to measure oestradiol production, and then processed for histological assessment of health status. Mean weight of ovaries and vaginal cul-de-sac tissues were both significantly greater (P<0.001) in FSH-treated anoestrous females (24.2+/-5.1 mg and 6.50+/-1.34 g, respectively), but did not differ significantly between saline-treated anoestrous possums (12.4+/-3.0 mg and 1.31+/-0.27 g) and cyclic animals (13.5+/-1.6 mg and 2.62+/-0.95 g). Mean uterine weights in both cyclic (889+/-161 mg) and FSH-treated (1098+/-184 mg) animals were significantly heavier(P<0.001) than those of anoestrous possums (414+/-61 mg). The mean number of follicles (> or =1.0-mm diameter) present was significantly greater (P<0.001) in FSH-treated, than in cyclic and anoestrous possums (38.0+/-4.4, 23.2+/-3.2 and 10.7+/-3.4 follicles/animal, respectively). Cyclic animals had significantly more (P<0.01) follicles than anoestrous possums. The proportion of follicles that were classified as healthy, was significantly lower (P<0.01) in cyclic possums(38%) than in anoestrous (69%) and FSH-treated (88%) animals. The mean diameter of the largest healthy follicle present was 2.5+/-0.41, 2.1+/-0.08, and 3.1+/-0.15 mm for cyclic, anoestrous and FSH-treated animals, respectively. None of the follicles harvested from saline-treated anoestrous possums produced measurable levels of oestradiol in vitro, whereas 7% and 59% of those from cyclic and FSH-treated animals did so. In summary, cyclic possums had more antral follicles present than anoestrous animals, but a lower percentage of these follicles were healthy. Less than 10% of healthy follicles from cyclic possums, and none of those from anoestrous animals, were capable of producing oestradiol when incubated in vitro. Treatment with ovine FSH promoted follicle development in anoestrous possums, to significantly increase the number of follicles present, the proportion that were healthy and the percentage capable of producing oestradiol.     

Ovarian follicular dynamics and hormonal profiles in heifer and mixed-parity Mediterranean Italian buffaloes (Bubalus bubalis) following an estrus synchronization protocol

The primary objective was to elucidate ovarian follicular dynamics and hormonal profiles in nulliparous heifer (HE; n = 11 ) and mixed-parity (MP; n=10 ) Mediterranean Italian water buffaloes (Bubalus bubalis) following an estrus synchronization protocol. Both groups received a progesterone releasing intravaginal device (PRID) implant for 10 days; a luteolytic dose of synthetic prostaglandin was given 7 days after PRID insertion. Daily ultrasound monitoring and collection of blood to determine plasma concentrations estradiol and progesterone started 1 day after PRID removal and lasted for 55 and 65 days in HE and MP buffaloes, respectively. Data analysis was restricted to the first 5 days after PRID removal and to one estrus cycle following induced ovulation. The HE buffaloes were not inseminated and only one ovulated within 5 days after PRID removal; the remainder ovulated between 8 and 48 days after PRID removal (except one in which ovulation was never detected). All HP buffaloes were inseminated 72, 96 and 120 h after PRID removal; seven buffaloes ovulated within 5 days after PRID removal and two were pregnant. Mean diameter of the largest follicle was significantly smaller in HE than MP buffaloes the first 4 days after PRID removal. There was a parity by time interaction ( P=0.0047 ) for plasma progesterone concentrations; progesterone was higher in HE than MP buffaloes 1 day after PRID removal, but the converse was true 2 days after PRID removal. After induced ovulation, HE buffaloes exhibited a one-wave ( n=5; length of cycle, 8-12 days), two-wave ( n=4; range: 20-26 days) or three-wave cycle ( n=1; 25 days). In contrast, all non-pregnant MP buffaloes ( n=8 ) had a two-wave cycle (range: 19-25 days). For buffaloes with two-wave cycles, the growth rate and diameter of the largest follicle was significantly smaller in HE than MP buffaloes for both the first follicular wave (1.3mm versus 1.7 mm per day and 10.5 mm versus 13.3 mm, respectively) and the second follicular wave (1.0 mm versus 1.3 mm per day and 11.0 mm versus 13.8 mm). In conclusion, there were many significant morphological and endocrine differences between HE and MP buffaloes.     

Comparison of two Ovsynch protocols (GnRH versus LH) for fixed timed insemination in buffalo (Bubalus bubalis)

We evaluated the efficiency of replacing GnRH with LH in the ovulation synchronization protocol in buffaloes. Buffaloes received GnRH on Day 0, (Buserelin; Conceptal, 20 microg), PGF2alpha (Luprostiol; Prosolvin, 15 mg) on Day 7 and GnRH (Buserelin; Conceptal, 10 microg; Group 1) or porcine LH (LH; Lutropin-V, 12.5 mg; Group 2) on Day 9. In Experiment 1, we studied the follicular dynamics of 30 buffaloes (Group 1, n = 15 and Group 2, n = 15). We performed ultrasonography every 12 h from Days 0 to 2, then on Day 7 and then every 6 h from the time of GnRH or LH treatment (Day 9) until the time of ovulation. All females not ovulating by 48 h after the second GnRH or LH injection were considered as nonresponders. In Experiment 2, we evaluated 305 buffaloes (Group 1, n = 154; Group 2, n = 151), using the same two treatments studied in Experiment 1. We also recorded and evaluated aspects like parity, lactational status, the presence of mucus, and uterine tone at the time of artificial insemination (Al). In Experiment 1, ovulation rate after the first GnRH was 86.6% (26/30). Ovulation rates were 93.3% (14/15; Group 1) after the second dose of GnRH and 93.3% (14/15) after LH (Group 2). Ovulation occurred 36.4+/-10.4 h after the first GnRH. The interval for treatment to ovulation was 26.5+/-9.6 h for buffaloes treated with GnRH (Group 1) and 24.4+/-7.9 h for buffaloes treated with LH (Group 2); the time of ovulation did not differ statistically between the two groups (GnRH versus LH; P > 0.05). In Experiment 2, conception rates of the animals AI in the field were 56.5% (Group 1) and 64.2% (Group 2), respectively (P = 0.08). The response to the treatment with LH was not different to the treatment with GnRH; however, multiparous buffaloes had higher conception rates than the primiparous buffaloes in both groups (P > 0.05). Buffaloes with mucus at the time of AI in Group 2 had higher conception rates than the buffaloes that had mucus in Group 1 (P < 0.05). Uterine tone and lactational status did not influence conception rates (P > 0.05). In summary, the results showed that both treatments resulted in synchronization of ovulation and acceptable conception rates. Therefore, the exogenous injection of LH can substitute the GnRH injections in the Ovsynch program in buffaloes.     

Evaluation of two different oestrus-synchronisation methods with timed artificial insemination and resynchronisation of returns to oestrus in lactating Holstein cows

To examine the outcomes of adding medroxyprogesterone acetate (MAP) to the ovsynch protocol with the traditional ovsynch protocol in both cycling and anoestrus cows, and to evaluate a resynchronisation protocol, 742 cows averaging more than 40 days postpartum were assigned to the following four treatments: (1) ovsynch (OVS): day 0: GnRH; day 7: PGF2alpha; day 9: a similar dose of GnRH; day 10: timed artificial insemination (TAI), approximately 16-20h later; (2) ovsynch+MAP (MAP): the same ovsynch protocol plus an intravaginal insert made of polyurethane sponge impregnated with 300mg of MAP immediately after the first GnRH treatment and on day 7, at the time of the PG treatment, the sponge was removed; (3) resynchronisation (MAP+ODB): 1mg of oestradiol benzoate (ODB) on day 13 after TAI and a new sponge impregnated with MAP was inserted and; on day 20, 1mg of ODB was given and the sponge removed; and (4) no resynchronisation (No MAP): only oestrus detection and AI at any repeat oestrus detected after TAI. Progesterone was measured in milk samples collected on days -17, -10, -3, 13 and 20 (TAI=day 0). Based on milk P4 at days -17 and -10, 27.4% of the cows were still anoestrus. At PG injection, 67.7% of the cycling and 21.3% of the anoestrus cows had elevated P4. Farm, days postpartum and parity variations were detected in both cases. On day 20 after TAI 42.6% of cycling and 8.3% of the anoestrous cows had elevated P4. Pregnancy rates were similar in both pre-breeding treatments (20%), but interactions (P<0.001) were detected between treatment and cycling activity (for anoestrous cows: MAP=34.9%; OVS=11.1%. Average interval from TAI to subsequent AI was 37+/-3 days. Resynchronisation resulted in more (P<0.001) cows in oestrus between days 18 and 25 after TAI. Conception rate in the MAP+ODB treatment was lower (P<0.05) than the No MAP group (22.8% versus 47.4%). It was concluded that the addition of a progestin to the ovsynch protocol resulted in increased pregnancy rates of cows treated during anoestrus. The benefit of including MAP with the ovsynch protocol for cycling cows is equivocal.     

The effect of eCG or estradiol at or after norgestomet removal on follicular dynamics, estrus and ovulation in early post-partum beef cows nursing calves

In post-partum anestrous beef cows suckling calves, neither the choice of hormonal regime to ensure the presence of a healthy dominant follicle at the end of a progestagen treatment nor the optimum hormone to induce estrus and ovulation is clear. Twenty-eight beef cows, in good body condition, 25-30 days post-partum, were assigned to one of four treatments: (i) 3mg norgestomet (N) implant with 5mg estradiol valerate (EDV) and 3mg N injection at the time of insertion (Crestar) for 5 days followed by 600 IU eCG at the time of implant removal; (ii) Crestar for 5 days as in (i) followed by 0.75 mg estradiol benzoate (EDB) 24h later; (iii) Crestar for 9 days followed by 600 IU eCG at the time of implant removal; and (iv) Crestar for 9 days followed by 0.75 mg EDB 24h later. Ovarian scanning was preformed from 4 days before implant insertion until ovulation and 4 days postovulation to detect the CL. Daily blood samples were collected from day 20 post-partum until second ovulation for FSH and E(2) assay. Data were analyzed using analysis of variance. There was no effect of the stage of follicle wave at the time of implant insertion on interval to new follicle wave emergence (range 1-7 days; mean 4.7 days). FSH concentrations were decreased to 5.9+/-2.0 and 7.7+/-1.1 ng/ml for pre- and post-selection cows 1 day after start of treatment; thereafter, they increased on Day 2 to 7.9+/-2.0 and 11.0+/-1.1 ng/ml and on Day 3 to 10.3+/-2.7 and 11.4+/-1.7 ng/ml for pre- and post-selection cows, respectively, despite high-estradiol concentrations at that time. There was no effect of treatment on the interval from implant removal to ovulation (3.2-4.0 days) or on the number of cows detected in estrus (26 of 27 cows). The size of the ovulatory follicle in cows given 0.75 mg EDB 24h post implant removal was decreased in animals at the pre-selection stage (12.2+/-0.1mm) of the follicle wave compared with those at the post-selection stage (15.3+/-0.9 mm) at implant removal. Cows given 600 IU eCG at the pre-selection phase of follicular growth had multiple ovulations (4.0+/-1.1). Cows given EDV at the start of a 5-day implant period had higher estradiol concentrations before and on the day of implant removal than those given EDV at the start of a 9-day implant period. The injection of 0.75 mg EDB 1 day after implant removal tended to increase concentrations of estradiol one day later. In conclusion, 5mg EDV and 3mg N at insertion of a 3mg N implant resulted in variable new follicle wave emergence 1-7 days later in post-partum beef cows nursing calves (22 of 27); both eCG and EDB were equally effective at inducing estrus after implant removal in cows in good BCS, but eCG resulted in a significant increase in ovulation rate in cows treated before dominant follicle selection.