Inability of progestogen pretreatment to prevent premature luteolysis of induced corpora lutea in the anestrous bitch

Fourteen mature anestrous bitches were used to determine the effectiveness of pretreatment with an orally active progestogen to prevent premature luteolysis of induced corpora lutea (CL) in the anestrous bitch. In Group 1, seven bitches were treated orally with megestrol acetate (Ovaban((R))) at the rate of 2.2 mg/kg body weight for eight days. Three days later, the bitches were treated daily with pregnant mare's serum gonadotropin (PMSG) (44 IU/kg body weight) administered intramuscularly for nine consecutive days, and each bitch was given 500 IU human chorionic gonadotropin (HCG) on day 10, or on the first day of induced estrus if the bitches exhibited estrus while being treated with PMSG. A control group (Group 2) of seven bitches was not treated with Ovaban((R)) but was similarly given PMSG and HCG. Estrus was detected twice daily using a vasectomized male dog and verified by vaginal cytology. Blood samples were obtained on the first day of induced estrus (day 0) and every other day until day 90 post-estrus. Plasma progesterone (P(4)) concentrations were determined by a non-extraction solid phase radioimmunoassay (RIA), and data were analyzed by Student's t-test. There was no significant difference between the progesterone profiles of both groups of bitches. In addition, P(4) values were less than 1 ng/ml by day 50 post-estrus. Results of this study suggested that pretreatment with an orally active progestogen was not effective in preventing premature luteolysis of induced CL in the anestrous bitch.     

Luteal function of induced corpora lutea in the bitch

Nineteen anestrous bitches with a mean of 22 kg body weight and ranging from 2 to 4 years of age were induced to exhibit estrus and ovulate using PMSG and HCG. Twelve days after the first day of estrus, bitches were assigned to four treatment groups. Group (A) consisted of six bitches, Group (B) of five bitches and Groups (C) and (D) of four bitches each. At this time, bitches in Groups (A), (B) and (C) were laparotomized and those assigned to Groups (A) and (B) were bilaterally hysterectomized leaving the cervix and oviducts intact. Although bitches in Group (C) were laparotomized, they were not hysterectomized. Group (D) bitches were not subjected to any surgical procedures. Homologous uterine extract was prepared from each bitch in Group (A) and administered intramuscularly beginning on day 25 (day 0 = first day of estrus) and continued every other day for 61 days post-estrus. Bitches in Group (B) were similarly injected with equal volumes of 0.9% saline. Blood samples, obtained prior to laparotomy and every other day for 85 days thereafter, were assayed for plasma progesterone concentrations using radioimmunoassay. One bitch in each of Groups (A) and (D) did not form luteal tissue following treatment with PMSG and HCG although both bitches exhibited estrus following treatment. All other bitches showed an increase in progesterone levels (4 to 19 ng/ml) between the first day of estrus and 10 days post-estrus. Thereafter, progesterone levels progressively declined in all groups with levels below 1 ng/ml between 38 to 40 days post-estrus. Results of this study suggested that CL formed in the bitch following PMSG and HCG treatment have a reduced function compared to non-induced CL of a normal, non-fertile estrous cycle. Such premature CL regression appears to be independent of the presence or absence of the uterus.     

Ovulation induction in anestrous bitches by pulsatile administration of gonadotropin-releasing hormone

Preliminary studies in anestrous Beagle bitches demonstrated that a single injection of gonadotropin-releasing hormone (150 micrograms) produced a rapid, physiological rise in serum estradiol lasting 1-3 days while progesterone remained below 1 ng/ml, whereas serial injections of FSH rapidly produced greater elevations in estradiol and a rapid rise in progesterone over 2 ng/ml. Consequently, attempts to induce fertile ovulation by means of pulsatile intravenous administration of GnRH (1 pulse/1.5 hours for 6-12 days; 0.04-0.43 micrograms/kg body weight/pulse) were conducted in eight anestrous bitches. Willingness to mate, serum progesterone levels and results of mating were monitored. In six of the eight bitches, vulval and vaginal signs of proestrus occurred by Day 2-4 after initiation of treatment (Day 0); but, two bitches showed negligible responses. In five of the six bitches in which proestrus was induced, behavioral (n = 4) and vaginal (n = 5) correlates of early estrus occurred by Day 5-7 of treatment and breedings occurred over a period of 4-12 days. Following onset of estrus, four of the five bitches had increases in serum progesterone levels between Days 14 and 18 after initiation of treatment (and 4-11 days after cessation of treatment); three of them became pregnant and whelped normal litters (ranging from 9 to 11 pups). The fifth bitch did not have elevated progesterone during the induced estrus, and upon return to estrus one month later was successfully bred and whelped a normal litter of 10 pups.(ABSTRACT TRUNCATED AT 250 WORDS)     

A model for cystic endometrial hyperplasia/pyometra complex in the bitch

The objective of this study was to develop a reliable model for the study of the cystic endometrial hyperplasia and pyometra complex (CEH/P) in the bitch. Greyhound bitches (n = 15) were ovariectomised and allocated into three groups (Group 1, n = 5; Group 2, n = 5; Group 3, n = 10, including 5 used from Group 1). Simulated proestrus, estrus and diestrus were induced by treatment with estradiol benzoate and megestrol acetate. The duration of cervical opening during estrus was determined by the intra-vaginal infusion of radio-opaque medium and subsequent radiography of the uterus (Group 1). One milliliter of a culture of Escherichia coli (with five uro-pathogenic virulence factors as identified by PCR: pap, sfa, hlyA, cnf1 and fim) was inoculated intra-vaginally daily throughout the simulated estrus (Group 2). One milliliter of the culture (n = 6) or sterile Luria-Bertani broth (n = 4) was introduced directly into the uterus on simulated diestrus Days 8 or 12 (Group 3). Necropsies were performed 12 and 7-14 days after the inoculation (Groups 2 and 3). The cervix remained open throughout the duration of simulated estrus (5-6 days) in four out of five bitches, and for a shorter duration (3 days of a 6-day estrus period) in one bitch (Group 1). CEH/P was induced by inoculation of bacteria into the uterus (10/10 bitches) but not into the vagina (0/5 bitches), (P = 0.003). A model for the study of CEH/P has been validated.     

Follicular development and plasma concentrations of LH and prolactin in anestrous female dogs treated with the dopamine agonist cabergoline

The effect of a daily administration of a dopamine agonist (cabergoline, 5 microg/kg) for 4 weeks, starting about 95 days after the end of estrus on follicular development and its relationship with LH and prolactin secretion has been investigated in two groups of anestrous bitches (Beagles and Greyhounds). Pro-estrus was detected in 80% (8/10) of beagles and 50% (3/6) of treated greyhounds. The mean inter-estrus interval of treated animals was 132+/-5.0 and 169+/-7.0 days for beagles and greyhounds, respectively, and in both this differed significantly from the cycle preceding treatment (192+/-9.0 and 198+/-12.0 days) and from that in untreated bitches (194+/-11.0 and 196+/-11.0 days for beagles and greyhounds, respectively (all comparisons at P<0.001). The interval from the beginning of treatment to pro-estrus in responding animals was 13.3+/-1.90 days in beagles and 20.3+/-1.70 days in greyhounds. Cabergoline increased (P<0.001) the length of pro-estrus (10.6+/-0.50 and 11.7+/-0.50 days) in the treated estrus cycle compared to the previous estrus cycle (8.4+/-0.30 and 8.8+/-0.40 days for in beagles and greyhound, respectively). Ovarian enlargement and follicle development was detected by ultrasound in 90% of treated beagles and in 83% of greyhound between the second and third weeks of treatment, but only 80% of beagles and 66% of treated greyhound displayed pro-estrus and estrus. In the treated bitches, mean plasma LH increased (P<0.001) before pro-estrus. There was high variability in mean plasma prolactin levels between animals. These data indicate that the administration of the dopamine agonist cabergoline to anestrous bitches increases mean LH plasma levels and induces follicular development shortly before pro-estrus but this activity is not always followed by pro-estrus and estrus. Finally, prolactin per se does not have a prominent role in the control of folliculogenesis in the bitch.     

Progesterone secretion by induced corpora lutea (CL) of anestrous bitches after administration of gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (HOG)

Sixteen bitches were treated with pregnant mare serum gonadotropin (PMSG; 44 IU/kg bwt) intramuscularly (i.m.) for nine consecutive days and each was given 500 IU HOG i.m. on the tenth day or on the first day of induced estrus (Day 0). On Day 12, each bitch was randomly assigned to three groups and treated as follows: Group 1-six bitches, each given 10 mcg GnRH i.m.once every 24 hr; Group 2-six bitches, each given 500 IU HOG i.m. once every 24 hr; and Group 3-four bitches, each given 5 ml 0.9% saline i.m. once every 24 hr. Treatments were continued until Day 55. Blood samples were obtained prior to treatment and every other day until Day 55. Plasma progesterone concentrations were determined by radioimmunoassay (RIA) and data arranged using a split-plot design, with treatment as the main plot and days of sampling as subplots. Analysis of data was by Duncan's Multiple Range Test and treatment-by-day interaction determined by the least-squares method. After treatment, progesterone concentrations from Group 2 were higher (P<0.05) than those of the other groups. A significant effect was seen on Day 20 (P<0.01), while on Day 50 the difference approached significance. These findings suggest that HOG is capable of stimulating the production of progesterone by induced CL in the bitch. However, neither HOG nor GnRH prevented premature regression of these induced CL. Results of this study suggest that premature luteolysis of induced CL is probably not due to lack of gonadotrophic support from either the hypothalamus or the anterior pituitary gland.     

The influence of exogenous progestin on the occurrence of proestrous or estrous signs, plasma concentrations of luteinizing hormone and estradiol in deslorelin (GnRH agonist) treated anestrous bitches

The objectives of this study were to confirm: (i) whether progestin treatment suppressed GnRH agonist-induced estrus in anestrous greyhound bitches; and (ii) the site of progestin action (i.e. pituitary, ovary). All bitches received a deslorelin implant on Day 0 and blood samples were taken from -1 h to +6 h. Five bitches were treated with megestrol acetate (2 mg/kg orally once daily) from -7 d to +6 d (Group 1) and 10 bitches were untreated controls (Group 2). Proestrous or estrous signs were observed in 4 of 5 bitches in Group 1, and 4 of 10 bitches in Group 2 (P = 0.28). The plasma LH responses (area under the curve from 0 to 6h after implantation) were higher (P = 0.008) in Group 2 than in Group 1. Plasma LH responses were similar (P = 0.59) in bitches showing signs of proestrus or estrus (responders) and in non-responders. The plasma estradiol responses (calculated as for LH response) were greater in Group 1 than in Group 2 (P = 0.048), and in responders than in non-responders (P = 0.02). In conclusion: (i) progestin treatment (a) did not suppress the incidence of bitches showing deslorelin-induced proestrus or estrus, and (b) was associated with a reduced pituitary responsiveness and an increased ovarian responsiveness to deslorelin treatment; (ii) the occurrence of proestrous or estrous signs reflected increased ovarian responsiveness to induced gonadotrophin secretion and not increased pituitary responsiveness to deslorelin.     

Fertile estrus induced in bitches by bromocryptine, a dopamine agonist: a clinical trial

The dopamine agonist bromocryptine, probably through amplifying gonadotroph (mainly FSH) secretion, was found to be suitable for provoking fertile estrus during the anestrous phase in bitches without functional cycles and/or ovarian activity. We studied estrus induction in 48 bitches after treatment with semisynthetic ergot alkaloid bromocryptine. For habituation a fractional dose of 0.3 mg/bitch was administered for three days followed by larger doses within the range of 0.6 to 2.5 mg/bitch by selecting dose rates on the basis of individual responsiveness and body weight. The long-term daily bromocryptine dose did not exceed 0.6 mg/bitch and 2.5 mg/bitch in small and large sized bitches, respectively. Gradual habituation and individual dose rates have almost completely eliminated the unwanted side effect of emesis. The period between treatment and onset of estrus varied but the average was 19 days. After the onset of estrus bromocryptine administration was usually continued for another 3 to 6 days. Occurrences of estrus, ovulation and pregnancy were monitored by cytological evaluation of vaginal epithelium, rapid ELISA for plasma progesterone and ultrasonography, respectively. Samples for progesterone were taken on Days 7, 9, 12 and 15 and sonograms of ovarian follicles and of fetuses were taken on Days 0, 22 and 35. The bitches involved in the study either regular or irregular cycles. Bromocryptine treatment induced estrus in all of the bitches including 40 of 48 (83%) with ovulation within a regular estrus and 6 of 48 (12.5%) that showed estrus but did not ovulate. Mating or artificial insemination of bitches in their fertile periods twice at two day intervals resulted in an 83% pregnancy rate (40 cases) and 39 (97.5%) of them gave birth to puppies. However, the average litter size was small with 4.8 +/- 1.6 pups.     

Pharmacokinetics of eCG and induction of fertile estrus in bitches using eCG followed by hCG

The aim was to design a protocol combining eCG followed by hCG for estrus induction in the bitch. In Experiment 1, three ovariohysterectomized bitches received 10 000 IU of eCG iv, and 15 days later 10 000 IU of eCG im. Blood samples were taken up to 144 h after each injection to measure eCG concentrations. In Experiment 2, 25 healthy, intact late anestrous bitches were assigned to one of five doses of eCG (5, 10, 15, 20, 44, or 50 IU/kg eCG im; [TRT5-TRT50]). Sexual behavior (SB), clinical signs of estrus (CSE) and vaginal cytology (VC) samples were obtained and scored before eCG administration and every other day until onset of estrus, or for 14 days. In Experiment 3, intact late anestrous bitches were assigned to a treatment group (TRT; n = 16) and received eCG (50 IU/kg im) followed by hCG (500 IU im) 7 days later; or to a placebo group (PLA; n = 8) where they received 1 mL saline solution im. All bitches that were induced in estrus were mated or AI with fresh semen. In Experiment 1, maximum observed concentration (C_max) eCG were similar between im and iv routes (6.1 ± 0.9 vs. 8.6 ± 0.5 IU/mL, P > 0.08), whereas time for maximum observed concentration (T_max.) was longer for im compared to iv routes (17.5 ± 0.5 vs. 11.6 ± 0.3 h, P < 0.01). The area under the curve (AUC) was similar for im and iv routes (P > 0.48), and eCG was detectable in serum for at least 144 h for both routes. In Experiment 2, 3 days or 3 to 5 days after treatment, all bitches in TRT50 had higher scores compared to TRT5-44 animals (P < 0.01). In TRT50, the mean interval from treatment to estrus was 4.0 ± 0.4 days. In Experiment 3, the mean interval from treatment to estrus was shorter in the TRT group compared to the PLA group (4.1 ± 3.3 vs. 68.5 ± 4.4 days, P < 0.01). The previous interestrus interval was similar for TRT and PLA groups (199.6 ± 7.2 vs. 197.5 ± 10.2 days), but the new interestrus interval was shorter for the TRT compared to the PLA group (164.0 ± 7.2 vs. 212.2 ± 10.2 days; treatment by interval interaction, P < 0.007). Serum P₄ concentrations increased on the first day of cytologic diestrus after treatment in bitches in TRT (0.7 ± 0.3 vs. 22.8 ± 4.2 ng/mL; P < 0.01); but did not change in PLA (P > 0.84). Ninety-four percent of animals were bred (15/16; AI, n = 7; natural mating, n = 8), and 80% (12/15) became pregnant. None of the bitches had any side effects from the eCG and hCG therapy. We concluded that 50 IU/kg of eCG combined 7 days later with 500 IU of hCG was effective to induce normal and fertile estrus in bitches at 164 days post estrus, with an 80% pregnancy rate, with no side effects, and with a reduction of 48 days of the interestrus interval.     

Induction of fertile estrus in bitches using a sustained-release formulation of a GnRH agonist (leuprolide acetate)

A single subcutaneous injection of a sustained-release formulation of a potent GnRH agonist, leuprolide acetate (LA; [D-Leu6, Pro9NEt]-GnRH), was evaluated as a method of inducing fertile estrus in 12 mature anestrous and 6 prepubertal beagle bitches. The bitches were treated with microencapsulated LA (100 micrograms/kg, s.c.) at 120 or 150 d post partum, or at 1 yr of age, followed by a GnRH-analogue (fertirelin; [Pro9NEt]-GnRH, 3 micrograms/kg, i.m.) on the first day of induced estrus. Signs of estrus were seen within 10.3 +/- 0.9 d after LA administration in all bitches. The interestrous interval in 120- and 150-d post-partum bitches was shortened (P < 0.05) to 191 +/- 3 and 222 +/- 3 d, respectively, compared with 264 +/- 11 d in control bitches. All LA treated dogs demonstrated behavioral estrus and mated. Three of 6 (50%) at 120 d post partum, 6 of 6 (100%) at 150 d post partum and 5 of 6 (83%) of prepubertal (1-yr old) bitches then became pregnant and produced a mean litter size of 4.1 +/- 0.8 pups. A normal circulating estrogen and progesterone response pattern was observed in mature anestrous bitches. A prepubertal bitch that failed to become pregnant had a similar estrogen response pattern but an insufficient progesterone profile. The results suggest that microencapsulated LA can be useful in inducing fertile estrus in the domestic dogs.     

Luteal function in the hysterectomized bitch following treatment with prostaglandin F2α (PGF2α)

Estrus and ovulation were induced in ten mature, mixed-breed, anestrous bitches (10 to 20 kg) using exogenous gonadotropins. Bitches were bred once, on the second day of estrus. Between 11 and 13 days following estrus, bitches were bilaterally hysterectomized and randomly divided into two treatment groups of five bitches each. Four days following surgery, Group A (treated) was given a single subcutaneous injection of PGF₂α (Prostin F₂ alpha^®) at a dose of 1 mg/kg body weight and Group B (controls) similarly given an equal volume of .9% saline. Blood samples were collected daily by cephalic venipuncture prior to surgery and for 75 days thereafter. Plasma progesterone was monitored by a radioimmunoassay method. Although bitches were teased daily following PGF₂α or saline treatments, estrual behavior was not exhibited. In both the PGF₂α and saline treatment groups, plasma progesterone levels showed a transient decline by 12 hours following injection, although a more dramatic decrease was observed at this time in the prostaglandin-treated bitches. Subsequently, progesterone concentrations tended to increase in both groups at 6 days following treatment, however, not to pre-treatment levels. Within 20 to 32 days following treatment in both groups, plasma progesterone levels declined to <1 ng/ml and remained depressed at least 60 days post-injection. In this study, complete luteal regression was not induced following PGF₂α treatment. Luteal function in both groups, as indicated by plasma progesterone concentrations, was shortened in the absence of the uterus.     

The effects of a low dose of cabergoline on induction of estrus and pregnancy rates in anestrous bitches

This is the first report of successful induction of normal estrus and ovulation in breeder bitches with as a low dose as 0.6 microg/kg/day of cabergoline formulation marketed for use in women. Sixty-one pure breed bitches from various breeds were used in the study at their already determined periods of anestrus. Twenty-four dogs formed the control group, while 37 bitches were administered with two different doses of cabergoline (recommended dose group, n=10, 5 microg/kg/day and low dose group, n=27, 0.6 microg/kg/day). Induced estrus rates and mean treatment and proestrus durations of dogs in these two dose groups were compared. At the second phase of the study, the effects of 500 IU human chorionic gonadotropin (hCG) administered on days 1 and 3 of estrus induced by the low dose of cabergoline, on the duration of behavioral estrus, ovulation rates, pregnancy rates and the number of offspring were investigated. For this purpose, the dogs with signs of proestrus (22/27) following the treatment in the low dose group were assigned into two subgroups. Five hundred IU of hCG (Pregnyl, Organon, Turkey) was intramuscularly administered to eight of these dogs [low dose (hCG+) group] on days 1 and 3 of estrus. The remaining 14 dogs were not treated with hCG [low dose (hCG-) group]. An aqueous solution of cabergoline (Dostinex, Pharmacia, Italy) was orally administered until 2 day after the onset of proestrus or for a maximum of 42 days. Blood samples were taken daily from all treatment and 11 control bitches during the first five days of behavioral estrus to measure progesterone concentrations. In the recommended dose and low dose groups, estrus was induced between days 8-45 and 4-48 (mean: 23.63+/-14.33 and 24.41+/-14.31 days), in the ratio of 80.0 and 81.5%, respectively (p>0.05). In both dose groups, post-treatment interestrous intervals were significantly shorter than both those of the control group and their own pre-treatment interestrous intervals (p<0.05). Ovulation rates, pregnancy rates and mean number of offspring delivered by the dogs in the recommended dose, low dose (hCG-), low dose (hCG+) and control groups were found to be similar (p>0.05). However, the mean duration of behavioral estrus of the dogs in the low dose (hCG+) group was found to be significantly longer compared to dogs in all other groups (p<0.05). In both dose groups, no correlation could be found between the anestrus stages and treatment durations (p>0.05). Shortly, it has been concluded from the study that (1) normal and fertile estrus can be induced more economically in bitches during different stages of anestrus using as a low dose of 0.6 microg/kg of cabergoline formulation marketed for use in women, and that (2) hCG injections on days 1 and 3 of the estrus induced by this method has no positive effects on the ovulation rates, pregnancy rates and the number of offspring per pregnancy.     

Induction of estrus in bitches with normal and persistent anestrus using human menopausal gonadotropin (hMG)

Human menopausal gonadotropin (hMG) was administered intramuscularly to 10 bitches during apparently normal anestrus (n = 7) or persistent anestrus (n = 3). Each dog received a 75-IU dose of hMG (75 IU LH and 75 IU FSH; 1 to 7 units/kg) daily for nine days. Nine bitches responded with obvious signs of proestrus within 3 to 9 days. Of these, 3 bitches exhibited a weak proestrus while 2 exhibited a normal estrus and ovulation but failed to become pregnant The remaining 4 bitches became pregnant at the induced cycle and produced normal litters at 72 to 85 d after the start of treatment, including 1 bitch that had been treated at 24 mo after the last estrus. In 2 cases, treatment resulted in ovulation following 25 or 34 mo of chronic pubertal anestrus, 1 of which became pregnant. The results suggest that hMG can be a useful gonadotropin preparation for inducing estrus in dogs.     

Effect of stage of anestrus on the induction of estrus by the dopamine agonist cabergoline in dogs

Beagle bitches were administered the dopamine D2 receptor agonist cabergoline in 3 groups of 5 animals each, starting on known days of the estrous cycle. Cabergoline treatment was started in either early anestrus (Days 93 to 108), mid-anestrus (Days 123 to 156), or late anestrus (Days 161 to 192) at doses of 5 ug/kg/d, per os, and was continued until the confirmation of induced proestrus or for 40 d. Reproductive parameters were compared with those in 5 control anestrous bitches (Days 90 to 150). In control bitches, the mean (+/- SEM) interval to the next proestrus (73+/-11 d) resulted in an interestrus interval (192+/-9 d) similar to that of the previous cycles (196+/-11 d). In 14 of the 15 cabergoline-treated bitches, the next proestrus occurred within 4 to 30 d, was premature in early and mid-anestrous bitches and developed with low variability within groups. The resulting intervals to proestrus in bitches treated with cabergoline in early anestrus (20+/-2 d), mid-anestrus (14+/-3 d) and late anestrus (6+/-1 d) resulted in interestrus intervals in those groups of 131+/-5, 166+/-7 and 196+/-2 d, respectively. In response to treatment, interestrus intervals were reduced (P<0.05) and more synchronous (P<0.05) in early and mid-anestrus bitches, and were more synchronous (P<0.05) in late-anestrous bitches compared with those of control bitches or those of the previous cycle. Periovulatory estradiol and progesterone profiles of induced cycles in treated bitches were similar to those of spontaneous cycles in control bitches. Four of 5 control bitches and 12 of the 14 responding cabergoline-treated bitches became pregnant and produced normal litters. Plasma prolactin concentrations at Days 2 and 5 of treatment (0.3+/-0.1 ng/mL) and at the onset of proestrus shortly before the end of treatment (0.4+/-0.1 ng/mL) were lower (P<0.05) than those present in anestrus prior to treatment (1.7+/-0.6 ng/mL) or in control bitches. Prolactin was also low at the onset of proestrus in control bitches (0.5+/-0.2 ng/mL). The results demonstrate that prolactin-lowering doses of the dopamine agonist cabergoline can terminate the normal obligate anestrus in dogs, and that the effect occurs more slowly in early anestrus than in mid or late anestrus.     

Postpartum subestrus in dairy cows: comparison of treatment with prostaglandin F2 alpha or GnRH + prostaglandin F2 alpha + GnRH

Two experiments (Experiment 1, 185 cows in 1996/97; Experiment 2, 168 cows in 1997/98) were conducted with Prim Holstein dairy cattle in the Mayenne region of France to investigate subestrus. Cows which had not been observed in estrus since calving were allocated alternately to treatment groups between 60 and 90 d post partum as follows: Experiment 1-Group 1: GnRH (Day 0, 100 micrograms i.m.), PGF2 alpha (Day 7, 25 mg i.m.), GnRH (Day 9, 100 micrograms i.m.) and AI (Day 10); Group 2: PGF2 alpha (Day 0, 25 mg i.m.), AI at estrus, or, if estrus was not observed, a second PGF2 alpha injection on Day 13, and AI on Day 16 and Day 17. Treatments in Experiment 2 were as follows: Group 1: as Experiment 1-Group 1 but AI at the observed estrus after Day 0, or at Day 10 if estrus was not observed; Group 2: as Experiment 1--Group 2, however, if a second PGF2 alpha injection was given on Day 13, AI at the observed estrus. Progesterone was measured in serum at Day 0 and in milk at AI. Pregnancy diagnosis was performed by measuring bovine pregnancy-specific protein B (bPSPB; Day 50 +/- 3) and confirmed by ultrasonography when the result was doubtful. In Experiment 1, farmers observed 47/101 (46.9%) Group 1 cows in estrus, 33/91 cows on Day 10 and 10 cows before Day 10. The progesterone concentrations were compatible with estrus in 69/86 (80%) cows on Day 10. In Group 2, 36/83 (43.4%) cows were inseminated after the first PGF2 alpha injection. After the second PGF2 alpha injection, only 29/43 (67%) cows had a low progesterone concentration at AI. Pregnancy rates were 36.1 and 32.5% for Groups 1 and 2, respectively. In Experiment 2, estrus was observed in 31/93 (33.7%) Group 1 cows. In Group 2, 51/75 (66%) cows were inseminated after the first injection of PGF2 alpha, 13/75 (17.3%) cows after the second injection, while 11/75 (14.7%) were not observed in estrus. Pregnancy rates were 53.7 and 53.3% in Groups 1 and 2, respectively. In conclusion, it is recommended that subestrus be treated with PGF2 alpha followed by AI at the observed estrus when estrus detection is good, while the use of GnRH + PGF2 alpha + GnRH is recommended when estrus detection is poor.     

Induction and synchronization of estrus in goats: the relative efficiency of one versus two fluorogestone acetate-impregnated vaginal sponges

The purpose of the experiment was to test the hypothesis that a variable and/or insufficient level of progestagen at the end of a treatment to synchronize estrus in goats could explain variability in the onset of estrus. The experiment was performed during the anestrous season on 2 herds, one of Alpine (n = 49) the other of Saanen (n = 53) dairy goats. The animals were allocated to 1 of 3 treatments: Group 1 received a vaginal sponge impregnated with 45 mg of fluorogestone acetate (FGA) on Day 0; Group 2 received a sponge on Day 0 plus a second sponge on Day 7; Group 3 received a sponge on Day 0 plus a second sponge on Day 9. The sponges were withdrawn on Day 11. All goats received 400 or 500 IU eCG and 50 mug PGF(2alpha) analog 48 h prior to sponge removal. They were inseminated with frozen-thawed semen 24 h after the onset of estrus. Among treatment groups no difference (P > 0.05) was observed for the following parameters: percentage of goats in estrus, percentage of goats ovulating, mean time and variability of onset of estrus. The fertility of Alpine goats in Group 3 was significantly decreased (P < 0.05). No effect on prolificacy was noticed. These observations show that to increase progestagen level at the end of treatment did not improve estrus synchronization. They provide further evidence that treatments with too high progestagen amounts can decrease fertility.     

Synchronization of estrus with PGF2 alpha administered 18 days after a progesterone treatment in lactating dairy cows

In a previous study we showed that estrus synchronization with 2 treatments of PGF2 alpha 13 d apart reduced conception rate at the synchronized estrus and that this reduction occurred mainly in cows in the early luteal phase at the second PGF2 alpha treatment. The objective of the present study was to determine the efficacy of a synchronization regimen in which PGF2 alpha was administered during the mid- to late-luteal phase to cows that had previously been synchronized with progesterone. Spring-calving cows from 6 dairy herds were used in this study. On Day -32 (Day 1 = the start of the breeding season), cows that had calved 2 or more weeks ago were randomly assigned to a synchronization (S, n = 732) or control (C, n = 731) group. Cows in Group S were treated with an intravaginal progesterone device (CIDR) for 12 d from Day -32 to Day -20, while those in Group C were left untreated. Similar percentages of cows in Group S (80.6%) and C (82.9%) had cycled by Day -7. The CIDR treatment synchronized the onset of estrus, resulting in 92.9% of cows in estrus being detected within 7 d after CIDR removal. Cows in Group S that had cycled by Day -7 were treated with PGF2 alpha (25 mg, i.m., Lutalyse) on Day -2. Cows in both groups that were anestrous on Day -7 were treated with a combination of progesterone and estradiol benzoate (EB) to induce estrus and ovulation (CIDR and a 10 mg EB capsule on Day -7, CIDR removal on Day -2, and injection of 1 mg EB 48 h after CIDR removal). The PGF2 alpha treatment synchronized the onset of estrus in 87.5% of the cows. Group S and C cows had similar conception rates to first (61.0 vs 58.3%) and second (58.4 vs 60.9%) AI; similar pregnancy rates over the AI period (82.8 vs 79.2%) and over the whole breeding season (91.9 vs 90.6%); and required a similar number of services per pregnancy to AI (1.7 vs 1.8). The interval from the start of the breeding season to conception for cows conceiving to AI or to combined AI and natural mating was shorter (P < 0.001) by 5.7 and 6.2 d, respectively, for the Group S cows. It is concluded that the treatment regimen tested in the present study achieved satisfactory estrus synchronization, had no detrimental effect on fertility at the synchronized estrus, and shortened the interval from start of the breeding season to conception.     

Induction and synchronization of estrus in dogs

Indications for estrus induction in the bitch include missed breeding opportunities or conception failure, the treatment of primary or secondary anestrus and synchronization of ovulation for embryo transfer programs. Reported methods for canine estrus induction include the use of synthetic estrogens (diethylstilbesterol), dopamine agonists (bromocryptine and cabergoline), GnRH agonists (lutrelin, buserelin, fertirelin, deslorelin, and leuprolide) and exogenous gonadotropins (luteinizing hormone, follicle stimulating hormone, human chorionic gonadotropin, pregnant mare serum gonadotropin, and human menopausal gonadotropin). These methods vary widely in efficacy of inducing estrus, as well as in the fertility of the induced estrus. The applicability of some of these methods for clinical practice is questionable. This review will summarize published reports on estrus induction and synchronization in bitches and summarize preliminary results using a long-acting injectable preparation of deslorelin.     

The regulation of precopulatory behavior by ovarian hormones in the female Mongolian gerbil

The hormonal regulation of precopulatory behavior in the female Mongolian gerbil was studied using two groups (N = 6) of sexually experienced females. A novel testing procedure was used which involved females living continuously with test males for several days. The test males showed either full sexual behavior (copulating males, C) or only precopulatory behavior (noncopulating males, NC). Experiment 1 investigated changes during the estrous cycle and following ovariectomy in females. Experiment 2 studied the effects of hormonal treatment of these ovariectomized females with 6 micrograms estradiol benzoate (EB) followed by 0.4 mg progesterone (P) or by 0.04 ml arachis oil. When tested with NC males, females displayed a greater range of precopulatory behavior. The patterns could be classified into three groups according to the manner of response to ovariectomy and hormone treatment. Group I patterns (approach, leave, and olfactory investigation of the male's head) were affected by neither ovariectomy nor EB treatment relative to Day 3 levels (Day 3, day preceding estrus; Day 4, estrus), but they were increased to estrous levels by EB and P. Group II patterns (darting, foot-stomping, and the present and piloerection postures) appeared only during estrus, did not appear after ovariectomy, and reappeared only after sequential EB and P treatment. Group III patterns (investigation of the male's anogenital area, allogrooming, ventral gland marking, and sand-rolling) were reduced relative to both estrus and Day 3 levels by ovariectomy and increased above Day 3 levels by EB alone; EB and P treatment further increased Group III patterns to the level of estrus. It is suggested that female precopulatory behavior patterns differ in their responsiveness to ovarian hormones. Estrogen appears to affect those patterns associated with the earliest stages of estrus (Group III).     

Combination dopamine agonist and prostaglandin agonist treatment of cystic endometrial hyperplasia-pyometra complex in the bitch

Cystic endometrial hyperplasia-pyometra (CEH-P) complex is a progesterone-dependent disease that requires medical treatment in bitches intended for breeding. To test the efficacy and safety of a combined protocol and to assess the effect of age, stage of cycle, previous steroid hormone administration and parity on treatment, 29 bitches diagnosed with CEH-P complex were treated daily with cabergoline 5 microg/kg PO and cloprostenol 1 microg/kg SC for 7-14 days, along with supportive antibiotic and hydration therapies. Before treatment, and on Days 3, 7 and 14, all bitches were evaluated clinically and uterine horn diameter measured during trans-abdominal ultrasonography. Twenty-four of 29 bitches were cured by either Day 7 or 14. Nine bitches had mild digestive side effects. Clinical signs related to pyometra began to improve markedly as early as Day 2 of treatment. Uterine diameters decreased (P < 0.05) by Day 3 of treatment, and continued to gradually decrease, reaching normal size by Day 14. Relapses occurred in 6 of 29 cases. Pregnancy was achieved in one of the two young bitches bred after treatment. No significant relationships were found between success rate and age, stage of the estrous cycle, previous hormone administration or parity. Although no variables affecting treatment results could be identified, this combination of compounds was found to be an efficient and safe for treatment of CEH-P.