A.C. Okkens

Termination of mid-gestation pregnancy in bitches with aglepristone, a progesterone receptor antagonist

Six pregnancies were terminated in mid-gestation with aglépristone, a progesterone receptor antagonist, in 5 beagle bitches in order to determine the effects of aglépristone on plasma concentrations of prolactin and progesterone, the duration of the luteal phase, and the interestrous interval. In addition, the effects of aglépristone on the condition of the uterus and fetuses were examined by ultrasonography. After confirmation of pregnancy by ultrasonography, the dogs received 10 mg, s.c. aglépristone per kg body weight on 2 consecutive days at about 30 d post ovulation. Before, during and after treatment with aglépristone, plasma samples were collected for determination of the concentrations of prolactin and progesterone. The condition of the uterus and fetuses was assessed by ultrasonography the day before and at least 3 times a week for at least 2 wk after aglépristone administration. Termination of pregnancy occurred within 4 to 7 d after the start of aglépristone treatment, which was well tolerated, with no side-effects except slight vaginal discharge. The results of ultrasonographic examination indicated that aglépristone leads to abortion but not to fetal resorption. Elevated plasma concentrations of prolactin were observed during aglépristone treatment, while plasma progesterone levels remained unchanged. Pregnancy termination with aglépristone resulted in premature cessation of luteal function. In addition, the interestrous interval was shortened. The latter effects may be the consequence of actions of the progesterone receptor antagonist at the hypothalamus-pituitary level. In conclusion, aglépristone proved to be a safe and effective abortifacient in mid-gestation in the bitch. The results of the present study also indicated that aglépristone directly or indirectly influences pituitary function.

Evidence for prolactin as the main luteotrophic factor in the cyclic dog

The role of prolactin and LH in the control of the function of the corpus luteum in the dog was studied. Experiments were performed to interfere with the secretion of a) prolactin by administering a dopamine agonist and b) LH by desensitisation with a long-acting LHRH and by stimulation. Treatments with prolactin-lowering dosages of bromocriptine, (20 micrograms/kg body weight twice a day, orally; n = 8) which started between day 1-5 (n = 4) and day 20-24 (n = 4) of the luteal period resulted in a similar pattern of progesterone, concentration in peripheral blood in both groups. The progesterone release in the second half of the luteal period (13.1 +/- 1.8% (sem) of the progesterone release of the total luteal period) was significantly lower than in control dogs (24.7 +/- 2.2%). Treatment at about day 30 of the luteal period with LHRH CR (1.34 mg, intramuscularly; n = 3), which significantly suppressed the LH level, did not reduce the progesterone release in the second half of the luteal period, 21.3 +/- 4.7% compared to 24.7 +/- 2.2% in the control dogs. The endogenous LH peak resulting from treatment with LHRH had no effect on the progesterone concentration in the blood. It is concluded that prolactin is the main luteotrophic factor in the cyclic dog during the second half of the luteal period.

Evidence for the non-involvement of the uterus in the lifespan of the corpus luteum in the cyclic dog.

Progesterone levels in peripheral blood of dogs were analysed during the cycle in which hysterectomy (n = 5) or sham surgery (n = 3) was performed as well as during the cycle of dogs (n = 5) hysterectomized at least one year prior to this study; the data were compared with the findings in control dogs (n = 3). The averages of the duration of the luteal period observed in the three experimental groups were not significantly different from those of control dogs. Immediately after surgery, the progesterone level decreased from 25 to 50% of the presurgical level, but returned to presurgical level in about four days. Prolactin levels were elevated for about 30 h after surgery. Nevertheless, the averages of the mean prolactin levels for each animal during the luteal period of the experimental groups were not significantly different from those of control dogs. It is concluded that in the dog, the uterus is not involved in the lifespan of the cyclic corpus luteum.

Shortening of the interoestrous interval and the lifespan of the corpus luteum of the cyclic dog by bromocryptine treatment.

Four beagle bitches were treated orally, twice daily with 250 micrograms bromocryptine, an inhibitor of prolactin secretion, from D1-D5 (D1 is defined as the first day of the luteal period) until the onset of the next prooestrus (n = 2) or the end of the luteal period (n = 2) of the following cycle. The mean interoestrous interval in the experimental group (123.3 +/- 23.1 day; n = 4) was significantly (p less than 0.001) shorter than the average of the mean values of the interoestrous interval (245.9 +/- 8.8 day; n = 36) of 10 control bitches. This shortening is mainly a consequence of a reduction to 35% of the anoestrous period of normally cyclic dogs. The luteal period of the first cycle was shortened to 78% compared with the luteal period of control dogs and this is also a contributing factor.

Influence of litter size and breed on variation in length of gestation in the dog.

The variation in the length of gestation, the period from mating until parturition, was studied in 77 dogs of different breeds; the time for mating was determined by measuring peripheral blood progesterone levels. The mean length of gestation was 62.1 +/- 0.2 (S.E.M.) days, with a variation of 11 days. The number of pups appeared to influence the length of gestation. Length of gestation was negatively correlated (r = -0.96, P < 0.001, n = 44) with litter size in litters with 7 or fewer pups. The intra-breed variation in length of gestation in the five breeds represented by five or more bitches was 3-6 days. The mean gestation of Alsatians (60.1 +/- 0.5, n = 9) was shorter (P < 0.005) than that of the other breeds combined (62.3 +/- 0.3, n = 68). The primiparous/multiparous status of the bitch did not influence the length of gestation.

Secretion of growth hormone and prolactin during progression of the luteal phase in healthy dogs: a review

In some dogs the long exposure to high circulating progesterone levels during each oestrous cycle may result in a syndrome of growth hormone (GH) excess, i.e. acromegaly. The progesterone-induced GH production in dogs with acromegaly originates from the mammary gland. Also in healthy cyclic bitches, the pulsatile secretion pattern of GH changes during progression of the luteal phase, with basal GH secretion being higher and pulsatile GH secretion being lower when plasma progesterone concentration is high. This may be explained by partial suppression of pituitary GH release by progesterone-induced GH production in the mammary gland. Progesterone also modulates the secretion of prolactin in the bitch. In pregnant and overtly pseudopregnant bitches, the plasma prolactin concentration starts to rise about 1 month after ovulation, which is when the plasma progesterone concentration begins to decline. Also in healthy cyclic bitches, most prolactin is released during the second half of the luteal phase. The changes in GH and prolactin release during the luteal phase may promote the physiological proliferation and differentiation of mammary gland tissue in the bitch. In the early part of the luteal phase progesterone-induced mammary GH initiates proliferation of the mammary epithelium, whereas in the late luteal phase, when progesterone concentrations decrease, prolactin release increases and promotes lobuloalveolar differentiation.

Low doses of bromocriptine shorten the interestrous interval in the bitch without lowering plasma prolactin concentration

In order to investigate the effect of different doses of bromocriptine on plasma prolactin concentration and the interestrous interval, beagle bitches were treated twice daily with 5 microg (5-group), 20 microg (20-group), or 50 microg (50-group) bromocriptine per kg body weight orally, starting 28 days after ovulation. In the 5-group, the difference between the mean plasma prolactin concentration before and that during bromocriptine treatment was not significant. In contrast, mean plasma prolactin concentration decreased significantly after the start of bromocriptine treatment in the 20- and 50-groups. The mean interestrous interval was significantly shorter in all three groups than in untreated bitches in the same colony. The mean interestrous interval in the 20-group and that in the 50-group were similar, but both were significantly shorter than that in the 5-group. The results of this study indicate that bromocriptine shortens the interestrous interval in the bitch even when the dose is so low that it does not lower plasma prolactin concentration. Induction of estrus in the bitch by bromocriptine therefore involves a mechanism other than via the lowering of plasma prolactin concentration. Furthermore, this study shows that the extent of shortening of the interestrous interval by bromocriptine is dose dependent.

Persistent Mullerian duct syndrome in a Miniature Schnauzer dog with signs of feminization and a Sertoli cell tumour

A 5-year-old male Miniature Schnauzer was presented with unilateral cryptorchidism and signs of feminization. Abdominal ultrasonography revealed an enlarged right testis and a large, fluid-filled cavity that appeared to arise from the prostate. Computed tomography revealed the cavity to be consistent with an enlarged uterine body, arising from the prostate, and showed two structures resembling uterine horns that terminated close to the adjacent testes. The dog had a normal male karyotype, 78 XY. Gonadohysterectomy was performed and both the surgical and the histological findings confirmed the presence of a uterus in this male animal, resulting in a diagnosis of persistent Mullerian duct syndrome (PMDS). The enlarged intra-abdominal testis contained a Sertoli cell tumour. Computed tomography proved to be an excellent diagnostic tool for PMDS.

Effects of gonadotropin-releasing hormone administration on the pituitary-gonadal axis in male and female dogs before and after gonadectomy

GnRH-stimulation tests were performed in 14 female and 14 male client-owned dogs of several breeds, before and 4 to 5 mo after gonadectomy. The aim of the study was to obtain more insight into the pituitary-gonadal axis in intact and neutered dogs and to establish reference values. Basal plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) concentrations were increased significantly after gonadectomy in both bitches and male dogs. In both males and females ranges of the basal plasma FSH concentrations, before and after gonadectomy, did not overlap as opposed to the overlap in ranges of the basal plasma LH concentrations. Before gonadectomy basal plasma LH concentrations were lower and basal plasma FSH concentrations were higher in bitches than in male dogs. After gonadectomy these basal values did not differ significantly. GnRH administration before gonadectomy resulted in an increase in plasma LH and FSH concentrations in both genders. GnRH administration after gonadectomy produced an increase only in plasma LH concentrations in both genders, and a just significant increase in plasma FSH in castrated male dogs. GnRH administration before gonadectomy resulted in a significant increase in plasma testosterone concentration in both genders. In males ranges of basal and GnRH-stimulated plasma testosterone concentrations before and after gonadectomy did not overlap. Basal plasma estradiol concentrations were significantly higher in intact males than in castrated males and their ranges did not overlap. The basal estradiol concentrations in bitches before and after ovariectomy were not significantly different. At 120 min after GnRH administration, ranges of plasma estradiol concentration of intact and ovariectomized bitches no longer overlapped. In conclusion, basal plasma FSH concentration appears to be more reliable than basal plasma LH concentration for verification of neuter status in both male and female dogs. The basal plasma testosterone concentration appears to be reliable for verification of neuter status in male dogs. The plasma estradiol concentration at 120 min after GnRH administration can be used to discriminate between bitches with and without functional ovarian tissue.

Fertility problems in the bitch

A short review of the oestrous cycle is given, followed by a more detailed look at fertility problems caused by husbandry aspects, anatomical abnormalities, hormonal disturbances and infectious aetiologies. One of the principal causes of fertility problems appears to be mating at an inappropriate time. The ovulation period and therefore the correct time of mating can however, be defined by measuring progesterone levels in peripheral blood three times a week. Vaginoscopy can also be used for this purpose, but is a subjective and less reliable method. Sterility is normally the single symptom of cystic endometrial hyperplasia(-mucometra). This probable major cause of sterility can sometimes be diagnosed with echoscopy but sometimes only during laparotomy. Additionally, hormonal disturbances can precipitate problems such as a persistent (pro-)oestrus, a split heat, or an abnormally long interoestrous interval. Bromocriptine, 15-20 micrograms/kg body-weight, twice daily orally, can be used to induce oestrus if no obvious abnormalities are diagnosed. Attention is paid to fertility problems with an infectious aetiology, such as herpes virus or Brucella canis. A vaginitis or abnormal bacterial growth in the vagina can also influence fertility. Unfortunately, little is known about bacterial growth in the vagina in relation to fertility problems. More research is needed in this field.