Efficacy of four density gradient separation media to remove erythrocytes and nonviable sperm from canine semen.

The objective was to compare four commercially available density gradient centrifugation (DGC) media (ISolate [Irvine Scientific; Santa Ana, CA, USA], Percoll [Pharmacia; Uppsala, Sweden], PureCeption [SAGE In-Vitro Fertilization, Inc.; Trumbull, CT, USA], PureSperm 100 [Nidacon International AB; Molndal, Sweden]) for their ability to separate viable, motile sperm from contaminant nonviable (immotile and/or dead) sperm and red blood cells (RBC). Pooled sperm-rich fractions from four healthy dogs were assessed using Spermvison SAR (Minitube of America). For this, 1 mL of the blood/sperm admixture was pipetted over 4 mL of DGC media: 50%/90% ISolate (Irvine Scientific), 45%/90% Percoll (Pharmacia), 40%/80% PureCeption (SAGE In-Vitro Fertilization, Inc.), and 40%/80% PureSperm 100 (Nidacon International AB). After centrifugation, five 1-mL fractions (A, B, C, D, and E) and the sperm pellet (bottom fraction F) were separated. Sperm morphology and red blood cell/sperm ratio (RBC/S) per fraction were determined on stained slides. All DGC media separated RBC from sperm; the highest red blood cell/sperm ratio was present in ISolate (Irvine Scientific) and Percoll (Pharmacia) fraction A (29.4±29.7 and 28.2±20.8, respectively), and in fractions A and B of both PureCeption (SAGE In-Vitro Fertilization, Inc.) (37.0±22.8 and 39.6±24.3, respectively) and PureSperm 100 (Nidacon International AB) (25.2±5.9 and 23.0±3.9, respectively). The fractions with the highest total sperm recovery, motile sperm recovery, as well as overall motility were ISolate (Irvine Scientific) and Percoll (Pharmacia) fraction D (33.9±29.4%; 40.99±27.9%; 71.2±21.8% and 36.4±14.5%; 39.3±15.8%; 88.6±2.3%, respectively), and for PureCeption (SAGE In-Vitro Fertilization, Inc.) and PureSperm 100 (Nidacon International AB), the sperm pellet, fraction F (78.8±28.3%; 88.0±17.4%; 70.2±11.1% and 73.1±21.0%; 75.4±24.6%; 80.6±17.1%, respectively). In the pellet for PureCeption (SAGE In-Vitro Fertilization, Inc.), more sperm and motile sperm were recovered than in ISolate (Irvine Scientific) and Percoll (Pharmacia) fractions D (P<0.0163). Therefore, DGC media should be considered for canine semen purification when contaminated with blood or when separation of motile versus immotile sperm is needed.

Retrograde ejaculation occurs in the dog, but can be prevented by pre-treatment with phenylpropanolamine: a urodynamic study.

Retrograde ejaculation is partial or total propulsion of semen from the posterior urethra into the urinary bladder; it is well characterized (and relatively common) in humans, with only a few reports in animals. Our objectives were to determine whether retrograde flow of semen occurred during ejaculation in mature dogs with normal fertility, and to determine the effects of phenylpropanolamine on this phenomenon (dose-titration, switch-back study). Retrograde ejaculation and urethral pressure profile measurements were evaluated (double-blind) in six dogs after 5 days of oral treatment with phenylpropanolamine (0, 2, 4, or 8 mg/kg); all dogs received all treatments (at 2-week intervals). The number of sperm in the urine was determined before and after each manual sperm collection. Urethral pressure profiles were obtained three times during each procedure. In the absence of phenylpropanolamine, sperm were present in the bladder after semen collection in all dogs (number varied significantly among individuals). The mean (+/-S.D.) number of sperm in the bladder was 17.0+/-5.0, 18.5+/-1.2, 5.1+/-5.0, and 4.8+/-0.1 x 10(6) sperm for 0, 2, 4, and 8 mg/kg, respectively (no significant difference between dogs given 4 or 8 mg/kg, but both were significantly lower than those given 0 or 2mg/kg). This reduction was significantly correlated to the increase in mean urethral pressure at the level of the sphincter (39 cm versus 59 cm H(2)O in placebo-treated dogs versus those given 8 mg/kg). In conclusion, we confirmed that retrograde ejaculation occurred during the ejaculatory process in normal dogs, and we demonstrated that phenylpropanolamine (4 or 8 mg/kg once daily for 5 days before collection) increased urethral pressure and reduced the number of sperm voided into the bladder during ejaculation.

Canine and feline pregnancy loss due to viral and non-infectious causes: a review.

Among the causes for pregnancy loss, viruses and non-infectious factors are among the most important. In both dogs and cats, research and clinical evidence provide proof that there is an increasing incidence of pregnancy loss associated with infectious diseases like herpesvirus, as well as the presence of toxicants or chemicals in the animal's diet and environment. Endocrine causes must be taken into consideration when dealing with pregnancy loss. This review will cover the most recent knowledge regarding viral and non-infectious of pregnancy losses in the dog and cat.

Endocrinology of pregnancy in the dog: a review.

Pregnancy regulation in the dog is not yet fully elucidated. Since plasma progesterone concentrations are similar in pregnant versus non-pregnant animals, it is a poor reflection on CL function and progesterone metabolism. Increased progesterone secretion by the CL in pregnant animals follows implantation and relaxin secretion by the feto-placental units. Progesterone is absolutely required to maintain pregnancy and no placental sources of progesterone have been identified. Pregnancy can be artificially maintained by progesterone administration. Prolactin secretion appears to be increased in response to the increase in relaxin production and occurs independent of estrogen production by the CL. The respective roles of LH, FSH and prolactin are still unclear, with considerable conflicting evidence among studies. However, it appears that prolactin is absolutely required, whereas LH is either permissive or facilitates CL function during pregnancy. Pre-implantation events are still poorly defined in the bitch, and no embryonic factors have been isolated or purified, preventing early pregnancy diagnosis. Parturition occurs following luteolysis, which results from the release of prostaglandin F(2alpha), which begins 36h prepartum in a process similar to that observed in other species. The role of estrogens at the time of parturition remains undefined.

Mucometra, cystic endometrial hyperplasia, and pyometra in the bitch: advances in treatment and assessment of future reproductive success.

Pyometra is a common reproductive disorder which affects nearly one fourth of all female dogs before they reach 10 y of age. An association between pyometra and the most common uterine disease of the bitch, cystic endometrial hyperplasia, has been established, as the latter allows commensal bacteria originating from the vagina to proliferate in the uterus at the end of estrus. The progressive degenerative process in the development of cystic endometrial hyperplasia is usually proposed as the initiating lesion for pyometra in bitches; this is mediated by progesterone and potentially aggravated by estrogens. However, a separate process caused by local uterine irritation to trophoblastic reaction and bacterial proliferation has been recently proposed as an alternate mechanism leading to the development of pyometra. Pyometra is clinically distinct in pathogenesis, signs, treatment and prognosis from postpartum metritis or mucometra. Treatment of pyometra has historically involved ovariohysterectomy, however, during the last 10 y, numerous effective treatments have been proposed to treat both open and closed cervix pyometra with good success and future fertility. Among the treatments available, the use of repeated low doses of prostaglandins alone or in association with either dopamine agonists or progesterone-receptor antagonists has been demonstrated to be a viable alternative for valuable breeding dogs.

Long-term motility and fertility conservation of chilled canine semen using egg yolk added Tris-glucose extender: in vitro and in vivo studies.

The effects of medium exchange on motility parameters of chilled canine semen preserved in egg yolk Tris-glucose (EYTG) extender were analyzed over a 27-d period. Semen extender was exchanged at three time points (Days 11, 21 and 27) after collection, when motility parameters were demonstrated to significantly decrease from parameters observed at semen preparation (Day 0) or at day of previous extender exchange. In the absence of medium exchanges, motile spermatozoa were observed up to Day 16 (mean +/- S.D. 1.5 +/- 0.3% of motile spermatozoa). A stimulation of the different semen motility parameters was observed after extender exchange. Semen extender exchange at Day 11 allowed conservation of motility until Day 21, compared to 16 d in the absence of extender exchange. At Day 21, when spermatozoa appeared immobile or dead, a second extender exchange was performed, allowing the extension of motility conservation up to Day 27. The third extender exchange, performed at Day 27, was no longer associated with motility stimulation. Glucose content in the medium decreased slowly over time; a concomitant decrease in pH was also observed. No changes in osmolarity were observed over time. To verify the fertility of long-term conserved chilled semen, two groups of 10 bitches were inseminated either once (Group 1) or twice at 48-h intervals (Group 2) intra-vaginally with semen conserved chilled for a mean of 9 +/- 1.8 d. Out of the 10 bitches inseminated once, 5 became pregnant, versus 7 in the group of animals inseminated twice. The present study reports the possibility to extend the conservation of chilled canine semen up to 3 wk with conservation of good fertility for at least 10 d. The role of energetic substrate and pH alteration is postulated and the classically accepted relation of semen motility/viability is raised.

Seroprevalence of canine herpesvirus-1 in the Belgian dog population in 2000.

Canine herpesvirus-1 (CHV-1) is known to be associated with fertility and fecundity disorders as well as neonatal mortality in puppies of less than 3 weeks of age. The virus is presumed to be enzootic in dogs all over the world and recent studies in several European countries suggest a high seroprevalence among the dog population. In the year 2000, a total of 647 Belgian canine sera from 102 privately owned patients and 545 breeding dogs were analysed with an enzyme-linked immunosorbent assay (ELISA). Furthermore 77 of the samples were submitted to two serum neutralization (SN) tests for comparison. An overall CHV-1 seroprevalence of 45.75% was observed in the Belgian dog population. No significant differences could be observed based on breeding status, reason for consultation or sex. The correlation between the ELISA and both SN tests appeared to be moderate with a significantly greater sensitivity of the ELISA. This study also demonstrated that the CHV-1 seroprevalence in the Belgian dog population is similar to that in other recently investigated European countries and that the incidence in breeding units is not necessarily higher than in non-breeding dogs.

Comparisons of estradiol, LH and FSH patterns in pregnant and nonpregnant beagle bitches.

To characterize plasma estradiol, LH and FSH patterns of secretion during the bitch estrous cycle, blood samples were obtained daily from 15 days before until 135 days after the LH surge in 10 pregnant and 10 nonpregnant beagle bitches. After an initial increase between days 15 and 10 and an expected proestrous peak, estradiol concentrations increased again from days 9-12 (corresponding to cytological metestrus) from basal values observed around day 9 after the LH surge, and remained significantly elevated throughout the luteal phase both in pregnant and nonpregnant animals. Concomitantly with the end of the luteal phase, plasma concentrations of estradiol returned to basal values in both groups. During the mid- to late-luteal phase, mean basal LH secretion was significantly elevated throughout in the pregnant relative to the nonpregnant animals. However, in nonpregnant animals, pulsatility was increased and peaks of higher amplitude were observed. The plasma FSH profiles, determined by a specific homologous RIA, differed significantly between pregnant and nonpregnant bitches during the last two-thirds of the luteal phase with a mean FSH level more elevated during pregnancy. The FSH level then decreased around parturition and low concentrations during lactation period were observed. The FSH concentrations remained steady in nonpregnant luteal phases from early luteal phase through mid-anestrus. The differences in pregnant and nonpregnant LH and FSH concentrations suggest pregnancy differences in regulation of the corpus luteum. Finally, the elevated estradiol concentrations observed during the luteal phase of both pregnant and nonpregnant animals suggest that an ovarian production of estrogens may be involved in overall corpus luteum regulation in dogs as in other species.

Computer assisted semen analyzers in andrology research and veterinary practice.

The evaluation of sperm cell motility and morphology is an essential parameter in the examination of sperm quality and in the establishment of correlations between sperm quality and fertility. Computer-assisted sperm analysis (CASA) allows an objective assessment of different cell characteristics: motion, velocity, and morphology. The development and problems related to this technology are raised in this review, paying particular attention to the biases and standardization requirements absolutely needed to obtain useful results. Although some interesting results, mainly in humans, have already been obtained, many questions remain, which have to be answered to allow for further development of this technology in veterinary medicine, clinical fertility settings, physiological, and toxicology research activities. The main problem is related to the standardization and optimization of the equipment and procedures. The different CASA instruments have all demonstrated high levels of precision and reliability using different sperm classification methodology. Their availability gives us a great tool to objectively compare sperm motility and morphology and to improve our knowledge and ability to manipulate spermatozoa.

FSH secretion patterns during pregnant and nonpregnant luteal periods and 24 h secretion patterns in male and female dogs.

The aim of this study was to investigate the secretion patterns of FSH: (i) during the canine oestrous cycle with particular attention to the luteal period; and (ii) during 24 h in male and female dogs. Plasma FSH concentrations were measured by a highly specific homologous canine immunoradiometrical (IRMA) assay using monoclonal antibodies specific for canine FSH. In the first study, FSH concentrations were measured daily from +/- 15 days before until 150 days after the LH surge in groups of pregnant (n = 5) and nonpregnant (n = 5) Beagle bitches. Day 0 of the cycle was the day of the preovulatory LH peak as measured by LH radioimmunoassay. In the second study, FSH concentrations were measured at 1 h intervals for 24 h in five bitches at day 120 after the LH surge and in five males. From day 35 to day 40 after the LH peak, FSH concentrations were higher in pregnant than in nonpregnant luteal phases. A significant and abrupt decrease in FSH concentrations was observed at about the time of parturition (day 65) and was followed by lower FSH concentrations during lactation compared with nonpregnant bitches. FSH concentrations in nonpregnant animals were fairly constant from the end of oestrus to mid-anoestrus. In both post-lactation and anoestrous bitches, plasma concentrations of FSH increased consistently as anoestrus progressed. No significant differences in the mean hour-to-hour patterns of FSH secretion were observed over 24 h and no differences were detected between male and female dogs. However, a clear pulsatile pattern of secretion was observed in all individuals, both males and females, with an apparent 4.8 h interval between peaks. The results of this study demonstrate major differences in FSH secretion between pregnant and nonpregnant bitches. This finding indicates that there are differences in the regulation of the hypothalamo-pituitary-ovarian axis during pregnancy, possibly related to changes in activity of the corpus luteum. The results of this study also demonstrate a pulsatile pattern of FSH secretion in both male and female bitches, with a 4.8 h interval between peaks, and confirms the progressive increase in plasma FSH concentrations observed during anoestrus.

Suppression of the oestrous responses of bitches to the GnRH analogue deslorelin by progestin.

Studies were undertaken in Australia and Belgium to determine whether the initial pro-oestrous-oestrous responses of anoestrous bitches to treatment with deslorelin administered in a s.c. implant were inhibited by progestin treatment. Thirty-nine bitches of mixed breeding were treated daily with 2 mg megestrol acetate kg-1 body weight for 21 (group 1, n = 5) or 14 days (group 2, n = 10), or with 1 mg megestrol acetate kg-1 body weight for 14 days (group 3, n = 10). A deslorelin (6 mg) implant was placed s.c. on day 14 (group 1) or day 7 (groups 2 and 3) of treatment. Bitches not treated with progestin also received a deslorelin implant (group 4, n = 9) or were untreated controls (group 5, n = 9). Signs of pro-oestrus-oestrus were not observed in bitches in groups 1, 2 and 5, but were observed in bitches in groups 3 (4/10) and 4 (9/9). Four bitches in group 4 were mated, two of which became pregnant. The pregnancies failed at about day 40 of gestation and were associated with low plasma progesterone concentrations. Treatment with progestin inhibited the pro-oestrous-oestrous responses of bitches to deslorelin.

Determination of the role of cervical closure in fertility regulation after mating or artificial insemination in beagle bitches.

In the present study, the pregnancy rate after one insemination with fresh semen performed at the optimum period of fertilization (control) was compared with the pregnancy rate after insemination by either vaginal deposition of semen or transcervical uterine insemination at 24, 48 or 72 h after closure of the cervix (n = 5 bitches per time period and group). In the group of bitches inseminated into the uterus, 2 ml of a fresh pool of semen from four male dogs was deposited transcervically or by laparoscopy into the uterus, whereas in the vaginally inseminated animals, 2 ml fresh semen was extended with 4 ml prostatic fluid. The study was performed over 5 years; some bitches were used twice during two consecutive oestrous cycles. Closure of the cervix was determined once a day by retrograde vaginogram using a radiopaque contrast dye. Closure of the cervix occurred 6.9 +/- 1.1 days (mean +/- SD) after the LH peak and 1.5 +/- 0.9 days before the first day of cytological metoestrus. The hormone values at the time of closure were 81.2 +/- 12.3 nmol progesterone l-1 and 28.4 +/- 23.2 pmol oestradiol l-1; there were no differences between groups. Of the bitches inseminated before closure of the cervix, five and four bitches became pregnant after uterine and vaginal insemination, respectively. Of the bitches inseminated into the vagina 24, 48 or 72 h after cervical closure, none became pregnant, whereas three, three and one bitches became pregnant after insemination directly into the uterus at 24, 48 or 72 h after cervical closure, respectively. The mean litter size was 3.9 +/- 1.4 pups among all pregnant animals and there were no significant differences between groups. The results of the present study indicate that mature canine oocytes may remain fertile for > 200 h after ovulation, rather than for 48-60 h as was previously believed, and that cervical closure may be a limiting factor for reproductive success after natural mating.

Time-related changes in canine luteal regulation: in vivo effects of LH on progesterone and prolactin during pregnancy.

The role of LH in luteal function in pregnant dogs was investigated at two different periods during pregnancy: (i) the transitional period from apparent total independence of the corpus luteum to relative hormonal dependence (days 20-35); and (ii) the period of full hormonal dependence (days 35-40). At both periods, LH neutralization, LH inhibition and LH administration studies were conducted. At both periods LH immunoneutralization had no significant effect on the secretion pattern of progesterone or prolactin. GnRH antagonist treatment (Nal-Glu) decreased plasma LH below the detection limit in all treatment periods. Nal-Glu had no effect on prolactin. When GnRH antagonist osmotic pumps were implanted, a transient decrease in plasma progesterone concentrations occurred on days 21-22 but not during the remaining implantation period. When GnRH antagonist was injected, plasma progesterone temporarily decreased (24 h) after the beginning of treatment starting on day 20, but decreased for 5 days when the treatment started on day 35. When purified pig LH was injected i.v. twice a day for 2 consecutive days either from day 30 or from day 40, plasma progesterone concentrations remained constant during treatment. However, on days 40 and 41, an increase in prolactin was observed. These results indicate that LH immunoneutralization may not impair corpus luteum function. In addition, GnRH antagonist induces dose- and time-dependent effects. Only high doses resulted in a decrease in progesterone, the duration of which increased as pregnancy progressed. Continuous GnRH antagonist administration, even when associated with complete LH inhibition, was not associated with detectable effects on progesterone. Finally, LH administration does not stimulate progesterone but may modify prolactin in the last third of pregnancy. Other studies indicated a corpus luteum prolactin dependency. The present study indicates that, in pregnant bitches, LH may not be necessary to sustain progesterone synthesis but that its role may vary in a time-dependent manner.

Comparisons of different combinations of analogues of PGF2 alpha and dopamine agonists for the termination of pregnancy in dogs.

Groups of five pregnant bitches were treated to terminate the pregnancy with four combinations of drugs, starting 28 days after the estimated surge of luteinising hormone (LH), 22 to 28 days after the first mating. The treatments were: cabergoline administered orally for 10 days at a dose of 5 micrograms/kg and a single subcutaneous injection of 2.5 micrograms/kg cloprostenol at the start of the treatment; the same dose of cabergoline plus two doses of 1 microgram/kg cloprostenol administered on days 28 and 32 after the LH surge; bromocryptine administered orally at a dose of 30 micrograms/kg three times a day for 10 days plus a single dose of 2.5 micrograms/kg cloprostenol; the same dose of bromocryptine plus two doses of 1 microgram/kg cloprostenol; and a group of five pregnant bitches was left untreated. The pregnancies were terminated in all but one of the treated bitches, in each case by resorption of the fetuses. There were few side effects in the bitches treated with two doses of 1 microgram/kg cloprostenol, and were present but acceptable in those treated with one dose of 2.5 micrograms/kg. Plasma progesterone concentrations decreased to less than 1 ng/ml within 72 hours of the start of treatment and remained low except in the bitch in which pregnancy was not terminated. In the five untreated bitches, plasma progesterone remained high and they whelped normally. In the treated groups, the intervals between successive displays of oestrus were reduced by approximately 70 days in comparison with previous cycles or with the control group, but the fertility of the dogs was not affected adversely.

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.

Termination of obligate anoestrus and induction of fertile ovarian cycles in dogs by administration of purified pig LH.

The potential role of LH in the initiation of the follicular phase in dogs was investigated by treating anoestrous bitches with highly purified pig LH (n = 16) or saline (n = 8) three times a day for 7 days, beginning in either early anoestrus (days 94-116 of the cycle) or mid-anoestrus (days 124-145). Treatment with LH induced pro-oestrus within 7 days (n = 16), and oestrus (n = 12) and fertile ovulations (n = 7) at 16 + 3 days, while pro-oestrus in bitches treated with saline did not occur until 46-166 days after the start of treatment. Six of the seven ovulating bitches whelped normal litters. The bitches in which pro-oestrus but not oestrus occurred were all treated in early anoestrus. During treatment, plasma oestradiol in bitches treated with LH increased from 8 + 2 pg ml-1 to 20 + 5 pg ml-1 within 1 day, and reached higher peak values (45 + 7 pg ml-1) (P < 0.05) than those observed in saline-treated controls (9 + 3 ng ml-1). These results demonstrate that LH treatment alone can terminate anoestrus by inducing a normal follicular phase in dogs. The results also suggest that, in normal cyclic bitches, anoestrus is the result of insufficient LH secretion, and that spontaneous pro-oestrus could be the result of increased LH secretion in the presence of already adequate concentrations of FSH.

In vivo investigation of luteal function in dogs: effects of cabergoline, a dopamine agonist, and prolactin on progesterone secretion during mid-pregnancy and -diestrus.

The role of prolactin on luteal function in dogs was investigated in vivo. The function of prolactin in mid-luteal phase was compared in pregnant and nonpregnant dogs. A dopamine agonist, cabergoline, known for its prolactin secretion inhibitory effects, was injected subcutaneously at a dose of 5 micrograms/kg body weight in five pregnant and five nonpregnant Beagle bitches. Mean plasma prolactin and progesterone were dramatically suppressed for 4 to 5 days after injection in both groups when compared with control pregnant and non-pregnant animals, whereas no effect on luteinizing hormone (LH) secretion was observed. The decline in plasma progesterone occurred after that in prolactin, suggesting plasma progesterone was impaired by inhibition of prolactin secretion. These results confirm the luteotropic importance of prolactin in pregnant bitches, and also demonstrate its importance in luteal phase of the nonpregnant dog. Second, to demonstrate that the effects of cabergoline were mediated by prolactin inhibition and not by a direct action on the corpus luteum, concomitant administration on Day 30 of cabergoline and prolactin (375 micrograms i.v. twice daily on Days 30 and 31) or cabergoline and LH (750 micrograms i.v. twice daily on Days 30 and 31) was affected in two groups of five pregnant animals each. Results showed that only prolactin was able to reverse the negative effects of cabergoline on circulating progesterone. This confirms the indirect mode of action of the dopamine agonist, cabergoline on corpus luteum function. Third, further investigation on the precise luteotropic role of prolactin was made by IV injection of 375 micrograms pure canine prolactin twice daily in five pregnant bitches on Days 30 and 31, and in five pregnant bitches on Days 40 and 41. No direct stimulatory effect of prolactin on plasma progesterone secretion occurred. Nor was there a noticeable effect on plasma LH secretion. These results suggest that prolactin is unable to directly stimulate progesterone secretion by the corpus luteum of pregnancy. The results of this study suggest that prolactin is an essential luteotropin in the dog from mid-luteal phase in both pregnant and nonpregnant animals. However, it appears to act by sustaining corpus luteum lifespan and function rather than by direct stimulatory effects on progesterone secretion.

Secretion patterns of plasma prolactin and progesterone in pregnant compared with nonpregnant dioestrous beagle bitches.

The plasma concentration of prolactin has been poorly described in dogs. Published data are still controversial. In this study, plasma prolactin and progesterone concentration patterns were determined in groups of ten pregnant and ten nonpregnant beagle bitches. Blood samples were collected daily between 08:30 h and 09:30 h from day-15 to day 135 after the LH peak. Plasma concentrations of prolactin increased from day 25 after the LH surge in the pregnant animals to a peak value on day 65, the mean day of parturition. After parturition, they fell for 24-48 h, and then increased again and remained high during lactation. Variations observed in lactation were great and probably due to suckling. In the nonpregnant bitches, plasma prolactin remained constant throughout the observation period. An increase was observed at about day 70, but was not significant. Plasma prolactin patterns differed greatly, depending on whether the bitch was pregnant or not, even though the luteal phases had the same apparent duration in both cases. No significant difference was observed between progesterone concentrations of pregnant and nonpregnant bitches, even if plasma progesterone was higher from day 40 in pregnant animals. The luteal period ended abruptly in pregnant bitches on day 64, that is 1 day before parturition, and was more progressive in nonpregnant animals. Significantly higher prolactin secretion in pregnancy suggests specific luteal regulation, whereas the lower concentrations observed in nonpregnant dioestrous bitches make its role in nonpregnant luteal phases questionable.

Termination of pregnancy in cats using a combination of cabergoline, a new dopamine agonist, and a synthetic PGF2 alpha, cloprostenol.

The objective of this study was to increase efficacy of treatment with the dopamine agonist, cabergoline, for inducing abortion in cats by combining it with a synthetic PGF2 alpha analogue, cloprostenol. Side effects of cloprostenol were avoided by using low doses. An oral formulation of cabergoline was chosen to facilitate administration. Cabergoline was given daily (5 micrograms kg-1 day-1) while s.c. injections of cloprostenol were administered every 2 days (5 micrograms kg-1) (2 days)-1). Plasma concentrations of progesterone, side effects and pregnancy outcome were compared with those of five untreated pregnant queens. The treatment was administered from day 30 after first mating in five queens confirmed as pregnant and lasted a mean of 11 +/- 1 days. All treated animals aborted in 9 +/- 1 days without any side effect, except a mild haemorrhagic vulvar discharge. Subsequent fertility of the queens was not compromised. Abortion was mediated by an abrupt and constant decrease in plasma concentrations of progesterone. Progesterone concentrations were lower than 1 ng ml-1 from day 38 onwards in all treated animals and remained at this value thereafter, except for one queen which showed a further increase in plasma progesterone concentration from day 44 without any clinical consequence. In conclusion, a combination of daily oral administration of cabergoline and cloprostenol injections every 2 days appears to be a reliable, safe and practical method for terminating pregnancy in cats at day 30 of pregnancy, when a diagnosis of pregnancy by palpation or ultrasonography can easily be made.

Practical use of a combination of a dopamine agonist and a synthetic prostaglandin analogue to terminate unwanted pregnancy in dogs.

The combination of a dopamine agonist, cabergoline, and a synthetic analogue of prostaglandin F2 alpha, cloprostenol, was used to induce termination of pregnancy from day 25 after the estimated luteinising hormone surge (day 27 +/- 4 after the first mating) in five mature beagle bitches. Cabergoline was administered orally at 5 micrograms/kg daily and cloprostenol was injected subcutaneously at 1 microgram/kg every other day. Treatment efficacy, in terms of pregnancy termination, was 100 per cent. Termination always took place by resorption of the fetuses. No side effects were observed. A mean of three injections of cloprostenol and nine days of cabergoline treatment was necessary to eliminate all signs of gestation. Termination was in each case accompanied by a decline in plasma progesterone (to less than 1 ng/ml) within 72 hours of initiation of treatment. In a control group of five, untreated bitches, plasma progesterone remained elevated throughout a corresponding period (from day 25 to day 50 after the estimated luteinising hormone surge). In the treated group, interoestrous intervals (98 +/- 41 days) were reduced, compared with previous cycles (194 +/- 9 days) or with those of the control animals (205 +/- 37 days). It is concluded that the combination treatment with the anti-prolactinic agent, cabergoline (5 micrograms/kg/day), and the synthetic prostaglandin F2 alpha, cloprostenol (1 microgram/kg/two days), is an easy, practical, reliable and safe method to terminate pregnancy near and before mid-gestation in dogs.