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.     

Short-term progestin treatments prevent estrous induction by a GnRH agonist implant in anestrous bitches

The objective of this study was to test the efficacy and safety of a short-term progestin treatment administered at two different times to prevent estrous induction in response to the administration of an implant releasing the GnRH agonist, deslorelin acetate (DA), in anestrous bitches. Interestrous intervals (IEI) observed prior to and post DA were compared. Forty-two anestrous bitches, with previous IEI history, were randomly allocated to one of the following treatments: PL: placebo sc (n = 12); MA: megestrol acetate 2mg/kg po for 8 days (n = 4); DA: 10mg sc (n = 8); MA&DA-1: MA beginning the day before DA (n = 8); and MA&DA-4: MA beginning 4 days before DA (n = 10). The dose of MA was identical for each treatment. All bitches were examined daily for 1 month and then every 3 months until the next spontaneous post-treatment estrous cycle. Post-GnRH estrous response occurred in 0, 0, 100, 50, and 10% of the PL, MA, DA, MA&DA-1, MA&DA-4, groups, respectively (<0.01). There was an interaction between the treatment and period for the duration of the IEI (< 0.01). Changes in IEI were different among treatments (p<0.01); the three DA-treated groups (147.5% +/- 10.3, 161.3% +/- 14.1, 148.6% +/- 19.2) differed from both the MA (12.9% +/- 17.6) and PL (8.1% +/- 7.8), but not among themselves. It is concluded that an 8 days megestrol protocol and DA on Day 4 was better than DA on Day 1 to prevent estrous response in anestrous bitches and that both protocols significantly increased the IEI.     

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.     

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.     

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 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.     

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.     

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.     

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.     

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.     

Estrus induction in the bitch using a combination diethylstilbestrol and FSH-P

The purpose of the study was to induce estrus and ovulation in normal bitches using a combination of diethylstilbestrol (DES) and follicle stimulating hormone of porcine pituitary origin (FSH-P). Thirteen mature mongrel female dogs were divided into two groups, the first group was treated for estrus induction during late anestrus and the second group during mid-anestrus. The dogs were monitored by teasing, vaginal cytology, and hormonal assay during the induced (n = 13) and the previous spontaneous estrous cycle (n = 9). Six of eight and three of five bitches came into standing estrus in the first and second group, respectively. Of the bitches that came into estrus, three conceived in the first group and one in the second. The average induced litter size was 7.0 versus 7.5 for the colony. Based on vaginal cytology the induced proestrus and estrus lasted 1.7 (0 to 3) and 12.9 (4 to 24) d, respectively, while the spontaneous proestrus and estrus lasted 5.8 (0-17) and 12.8 (9-15) d, respectively. Progesterone profiles were similar between the induced and spontaneous estrous cycles, although the progesterone peak was higher during the spontaneous cycle. The preovulatory luteinizing hormone (LH) surge was observed in only one induced estrous cycle. Modest results were obtained with this therapy. However, the litter sizes were normal and the induced cycles were very similar to the physiologic ones. No side effects were seen with the oral form of DES.     

Reproductive cycles and contraception of black lemurs (Eulemur macaco macaco) with depot medroxyprogesterone acetate during the breeding season

Contraception is a critical component of population management for lemurs, but concerns about potential deleterious effects of continuous, long‐term treatment with synthetic progestins such as the widely used melengestrol acetate implant led us to evaluate seasonal contraception with injections of depot medroxyprogesterone acetate (Depo‐Provera, Upjohn Pharmacia, Kalamazoo, MI) to limit the duration of exposure. We compared two dosage regimens in female black lemurs using vaginal cytology as an indirect measure of ovarian suppression. Our results indicate that both 10 mg/kg body weight at 90‐day intervals or 2.5 mg/kg at approximately 30‐day intervals can be effective in most females, although one female on the 10‐mg dose showed signs of estrus at 53 days. Darkening of pelage during treatment was the primary side effect noted. A more important observation was that contraception can extend the breeding season to as much as 9 months, considerably longer than reported previously, which necessitates extending the period of contraceptive treatment. Zoo Biol 26:289–298, 2007. © 2007 Wiley‐Liss, Inc.     

Long-term melatonin treatment prolongs interestrus, but does not delay puberty, in domestic cats

The objective was to assess the efficacy and safety of long-term administration of melatonin (either as an implant or given orally) on interestrus intervals in domestic cats. Additionally, the effect of melatonin implants on puberty postponement was studied. For these purposes, two randomized controlled trials were conducted. In the first, 68 interestrus intervals (in 28 postpubertal queens) were studied, and in the second, 32 prepubertal female cats were used. During anovulatory interestrus intervals (27 ovulatory interestrus intervals were excluded), postpubertal cats were assigned to the following three treatments: melatonin implant 18 mg/cat SC (n = 17; MEI); melatonin tablets, 4 mg/cat/d orally until the onset of estrus (n = 12; MEO); or control (n = 12; CTL). Prepubertal females were randomly assigned to the following three treatments: melatonin 18 mg/cat sc implants at either 1.9 ± 0.3 kg (MEI-A; n = 12) or 1.5 ± 0.3 kg (MEI-B; n = 10) body weight; or control (CTL; n = 10). Interestrus intervals in postpubertal MEI, MEO, and CTL groups were 63.8 ± 5.4, 63.0 ± 5.3 and 19.2 ± 1.4 d (P < 0.05), respectively. In these groups, intervals between onset of treatment and the first estrus cycle were 51.0 ± 4.7, 50.0 ± 6.1, and 12.6 ± 1.1 d (P < 0.05). In the second experiment, neither age (MEI-A: 232.4 ± 10.5, MEI-B: 208.6 ± 13.0 and CTL: 192.4 ± 20.1 d; P > 0.1) nor body weight (P > 0.1) at puberty differed among groups. None of the cats in either study had clinically apparent side effects. We concluded that long-term melatonin treatment of domestic cats slightly prolonged interestrus intervals, but did not postpone puberty.     

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.     

Selective control of the estrous cycle of the dog through suppression of estrus and reduction of the length of anestrus

The objective of this study was to determine the effectiveness of testosterone in suppressing estrus in the bitch, and of cabergoline in shortening the length of the subsequent anestrous period. In Experiment 1, 12 diestrual Beagle bitches were randomly divided into two groups when plasma progesterone (P(4)) concentration was <1 ng/ml (Day 0). Starting on Day 0, bitches in Group 1 (n=6) were treated with testosterone cypionate every 14 days for a total of 239 days, and bitches in Group 2 served as untreated controls. On Day 274, bitches in both groups were treated with cabergoline for 40 days and blood samples were obtained on Days 274, 276 and 279 for determination of plasma prolactin (PRL) concentrations using RIA. All bitches were observed for proestrual bleeding during treatment with cabergoline. In Experiment 2, 12 Greyhound bitches previously treated with testosterone within the last 6 months were randomly divided into two groups. At the initiation of this experiment, P(4) concentration was determined to verify that all bitches had a concentration of <1 ng/ml (Day 0). Starting on Day 0, bitches in Group 1 (n=6) were treated with cabergoline for 36 days, and bitches in Group 2 (n=6) served as untreated controls. Blood samples were obtained on Days 0, 2 and 5 to determine PRL concentrations. All bitches were observed for proestrual bleeding during treatment with cabergoline. In Experiment 1, one bitch (Group 1) exhibited estrus after treatment with testosterone (1mg/kg body weight) for 43 days, and one bitch (Group 1) exhibited estrus after treatment with testosterone (2mg/kg body weight) for 113 days. None of the other four bitches in Group 1 exhibited estrus during the period of testosterone treatment (239 days). All bitches in Group 2 (control) exhibited estrus during the 239 days of the study. In addition, five of the six testosterone-treated bitches showed signs of proestrual bleeding within an average of 12.6 days (range of 5-25 days) after treatment with cabergoline; and, four of the six nontestosterone bitches showed signs of proestrual bleeding within an average of 28 days (range of 6-46 days). Prolactin concentrations in bitches in both Groups 1 and 2 significantly decreased after treatment with cabergoline. In Experiment 2, one of the six bitches showed signs of proestrual bleeding within 15 days after treatment with cabergoline. From the results of this study, it was concluded that exogenous testosterone was moderately effective (66%) in suppressing estrus in Beagle bitches, and cabergoline was effective in shortening the length of the anestrous period of Beagle bitches whose estrous cycle was previously suppressed with exogenous testosterone, but less effective in shortening the length of the anestrous period in Greyhound bitches previously treated with testosterone to suppress estrus.     

Effect of ovarian antral follicle cauterization on the interestrus interval of the gilt

The objective of the present study was to determine if destruction of ovarian antral follicles by laser-cauterization affects CL lifespan during the estrous cycle of the gilt. Cyclic gilts were randomly assigned to either SHAM, laser (L) or laser-estradiol (L-E2) treatment groups, with the L-E2 group receiving a 5-mg intramuscular (i.m.) injection of estradiol-17beta cypionate at the time of the first surgery. Ovarian antral follicles were laser-cauterized on either Days 12 and 14 (L12) or Days 14 and 17 (L14) of the estrous cycle. In the L12-E2 group, 3 of 4 gilts had extended mean interestrus intervals of more than 22 days compared with 0 of 4, 0 of 6, 0 of 7 and 1 of 5 gilts in the SHAM, L12, L14 and L14-E2 groups, respectively. The L12-E2 gilts had a longer (P<0.05) mean interestrus interval (23.5+/-1.3 days) than the L12 (20.0+/-1.1 days), L14 (20.7+/-1.0 days) and SHAM (20.5+/-1.3 days). The mean interestrus interval of L14-E2 gilts (21.8+/-1.2 days) did not differ from those of the L12-E2 group or the L12, L14 and SHAM group gilts. Six additional gilts were injected with 5 mg estradiol cypionate-17beta to serve as nonsurgical controls for E2 treatment. Gilts (3 of 3) given an E2 injection on Day 12 had extended mean interestrus interval (26.0+/-2.6 days), while 2 of 3 gilts injected with E2 on day 14 had extended mean interestrus intervals (27.7+/-2.1 days). These results indicate that in cyclic gilts destruction of ovarian follicles by laser-cauterization did not affect CL lifespan, and that luteolysis is not dependent on the presence of antral follicles.     

Prolactin receptors in the rat mammary gland. Change during the estrous cycle

Ovine prolactin (o-PRL) binding to mammary gland membranes was studied during the estrous cycle in the rat. Groups of rats were decapitated throughout the 4-day estrous cycle at 10 h00 on the days of diestrus I, diestrus II and estrus and at 10 h00, 12 h00, 16 h00 during the day of proestrus. Daily vaginal smears were taken to determine the stage of the estrous cycle which was also controlled by PRL and LH serum levels. Prolactin receptors were quantified in the 100 000 g pellet. For one Scatchard analysis, mammary gland membranes from 5 animals were pooled. Results given are the mean of 4 or 5 pools. Results obtained showed that the apparent affinity constant (KA) remained unchanged during the days of diestrus II and at all the times studied of proestrus and showed a slight but significant decrease on the days of estrus and diestrus I (or metestrus). The binding capacity did not vary from the day of diestrus II to the proestrus 16h00 (11.3 +/- 2.8 fmoles/mg protein) but sharply increased on the day of estrus (190.4 +/- 35.9 fmoles/mg protein). Binding capacity remained elevated on the day of diestrus I. This increase of PRL receptors on the day of estrous would appear to be an important step in preparing mammary gland for pregnancy and lactation.     

Effects of dose and duration of continuous GnRH-agonist treatment on induction of estrus in beagle dogs: competing and concurrent up-regulation and down-regulation of LH release

Dose-response estrus-induction trials were conducted during anestrus in 93 treated and 6 control bitches, a continuous administration of the GnRH-agonist lutrelin with a potency 150 x GnRH, and at six different doses from 0.2 to 4.8 microg/kg/d for 7-14 days in 15 groups of six to eight dogs each in defined stages of natural or pharmacologically determined anestrus. Agonist treatment induced clinically and cytologically normal proestrus (in 89% of cases) within 4.8 +/- 0.2 x days, and resulted in behavioral estrus (71%), spontaneous late-proestrus LH (and FSH) surges, ovulation (59%) and pregnancy (44%) in a dose dependent manner. Outcomes of ovulation and pregnancy in most cases required that the dose be sufficiently large enough to routinely stimulate a large initial increase in LH and FSH (i.e., > or = 0.6 microg/kg/d), and of sufficient duration (i.e., > 7 days) to ensure that supra-basal gonadotropin levels persistedntil no longer needed for spontaneous continuation of an induced proestrus. Success additionally required that the GnRH dose be modest enough (i.e., < 1.8 microg/kg/d) to not excessively down-regulate spontaneous pre-ovulatory surge release of gonadotropin or be removed shortly before or at the time when the LH surges typically occurred (10-13 days after initiation of treatment). The 1.8 microg dose was compared to saline to assess the time course of its down-regulation action on serum LH in six ovariohysterectomized bitches compared to four saline-related controls. Results in intact bitches receiving the 1.8-microg doses demonstrated an LH-releasing effect for 10-11 days that overlapped a period of obvious down-regulation seen with the same dose after 3 days in the ovariohysterectomized bitches. In the latter, however, complete down-regulation to anestrus-like values did not occur until after 18-21 days of treatment. A dose of 0.6 microg/kg/d for 12 days yielded the best estrus-induction results, including pregnancy rates of 100% in six bitches treated in natural-anestrus bitches, six bitches in which anestrus had been advanced by a luteolytic prostaglandin treatment and in six bitches in which anestrus had been extended by progesterone implants administered for 3 months. Although lutrelin is not commercially available, these results provide guidelines for the development of estrus-inducing protocols with other GnRH-agonists of known biopotencies.     

Treatment with a subcutaneous GnRH agonist containing controlled release device reversibly prevents puberty in bitches

In most species, continuous administration of GnRH agonists desensitizes the pituitary to GnRH, and blocks ovarian function. The aim of this study was to assess the effects of a novel controlled release device containing azagly-nafarelin (Gonazon) to prevent puberty in young Beagle bitches (mean age: 4.88 +/- 0.32 months). Gonazon containing 18.5 mg azagly-nafarelin (n = 10) or a placebo implant (n = 10) was administered subcutaneously. Throughout the 1-year treatment, estrus behaviour was monitored weekly. Plasma progesterone concentrations, as well as body weight and height, were measured monthly. Following implant removal, estrus detection and progesterone measurement were continued until occurrence of puberty in all bitches. Control bitches displayed puberty (estrus, followed by ovulation) at approximately 11.9 +/- 2.7 (range, 8-16) months of age. In contrast, none of the Gonazon treated bitches displayed puberty during the period when Gonazon was present. Following removal of Gonazon, resumption of estrus and ovulation naturally occurred (seven bitches) or was induced (three bitches) approximately 8.5 (1.2-14.3) months later. As a consequence, age of puberty of the Gonazon treated bitches was 25.5 +/- 5 (18-31) months. No clinically detectable side effects were noted in Gonazon treated bitches. Height at withers was unaffected by treatment. Changes in body weight with time were also unaffected by treatment. Implants were well tolerated and generally easy to remove. These data demonstrated that Gonazon safely, efficiently and reversibly prevents reproductive function for 1 year in prepubertal bitches.