The function of the pituitary-testicular axis in dogs prior to and following surgical or chemical castration with the GnRH-agonist deslorelin.

Chemical castration, that is the reduction of circulating testosterone concentrations to castrate levels by administration of a GnRH-agonist implant, is a popular alternative to surgical castration in male dogs. Detailed information concerning the pituitary-testicular axis following administration of a GnRH-agonist implant is still scarce. Therefore, GnRH-stimulation tests were performed in male dogs, prior to and after surgical and chemical castration. This approach also allowed us to determine plasma concentrations of testosterone and oestradiol in intact male dogs for future reference and to directly compare the effects of surgical and chemical castration on the pituitary-testicular axis. In intact male dogs (n = 42) of different breeds GnRH administration induced increased plasma LH, FSH, oestradiol and testosterone concentrations. After surgical castration basal and GnRH-induced plasma FSH and LH concentrations increased pronouncedly. Additionally, basal and GnRH-induced plasma oestradiol and testosterone concentrations decreased after surgical castration. After chemical castration, with a slow-release implant containing the GnRH-agonist deslorelin, plasma LH and FSH concentrations were lower than prior to castration and lower compared with the same interval after surgical castration. Consequently, plasma oestradiol and testosterone concentrations were lowered to values similar to those after surgical castration. GnRH administration to the chemically castrated male dogs induced a significant increase in the plasma concentrations of LH, but not of FSH. In conclusion, after administration of the deslorelin implant, the plasma concentrations of oestradiol and testosterone did not differ significantly from the surgically castrated animals. After GnRH-stimulation, none of the dogs went to pre-treatment testosterone levels. However, at the moment of assessment at 4,4 months (mean 133 days ± SEM 4 days), the pituitary gonadotrophs were responsive to GnRH in implanted dogs. The increase of LH, but not of FSH, following GnRH administration indicates a differential regulation of the release of these gonadotrophins, which needs to be considered when GnRH-stimulation tests are performed in implanted dogs.

Case Report: Medical Management of Prolonged Gestation of a Mummified Fetus in a Bitch.

A 3-year-old female rough coated collie was presented at day 69 (D69) after the first mating. She was mated on 2 consecutive days based on ovulation timing by the referring veterinarian. At day 30 post breeding, a single, live embryo was seen on ultrasound by this veterinarian. On D69, the bitch was presented to us because she lacked signs of impending parturition such as vulvar discharge or nest building behavior. On general examination, the bitch appeared clinically healthy and no prodromi were present. On abdominal palpation a small, firm structure and a slightly enlarged uterus were detected. There was no vulvar discharge. Using vaginoscopy we could not see any signs of cervical dilatation. Additionally, ultrasonography revealed the presence of a collapsed fetus in the uterus with a moderate amount of echogenic fluid surrounding it and the plasma progesterone concentration was 2.6 ng/ml. A parturition induction protocol was initiated: a progesterone receptor antagonist was administered, followed by PGF2α to induce cervical relaxation and uterine contractions. The fetus was expelled 3 days later, without noticeable damage to the reproductive tract of the dam. The bitch subsequently delivered two more litters without complications. To our knowledge this is the first clinical report that demonstrates a successful non-surgical treatment to expel a mummified fetus after prolonged gestation. The pharmacological treatment did not affect the future fertility of the breeding dog, which is an important outcome for breeders.

A laparoscopic approach for removal of ovarian remnant tissue in 32 dogs.

BACKGROUND: Surgical treatment of ovarian remnant syndrome (ORS) in dogs usually necessitates large celiotomies and considerable manipulation of organs because of the relatively deep position of ovarian remnant tissue, large patient size, and often encountered adhesions. In women, laparoscopic treatment of ORS is successful and has significant advantages over laparotomy. Since laparoscopic ovariectomy has significant advantages over open ovariectomy in dogs, including reduced surgical stress and postoperative pain and shorter convalescence period, the rationale for a laparoscopic approach of canine ORS is evident. Feasibility and efficacy of a laparoscopic approach for treatment of ORS in dogs was prospectively evaluated using a standardized protocol for diagnosis, treatment, and follow-up. Treatment success was evaluated by histology of removed tissues, postoperative hormone testing, and long-term clinical follow-up. RESULTS: Thirty-two client-owned predominantly medium and large breed dogs diagnosed with ORS underwent abdominal ultrasound for ovarian remnant localization prior to laparoscopic surgery for removal of ovarian remnants. Tissue dissection and excision was performed using a vessel sealing forceps. Laparoscopy subjectively enabled detailed visibility and facilitated detection and removal of suspected ovarian tissue in all cases. Histology confirmed ovarian origin of removed tissue in all dogs. Additionally, a GnRH stimulation test was performed in fourteen dogs after a median follow-up of 10.5 months, which verified absence of residual functional ovarian remnant tissue in all dogs. Median surgery duration was 97.5 min and mean total convalescence duration, subjectively scored by owners, was 1.5 ± 0.7 days. No major complications occurred. Adhesions were observed in 79% of the dogs, complicated the surgical approach, and significantly affected surgery duration (85 versus 109 min; p = 0.03). Minor hemorrhage occurred in 12% and significantly increased surgery duration (95.5 versus 128 min; p = 0.02). Trendelenburg position and lateral tilting of the patient were essential for proper access to ovarian remnants. GnRH stimulation test results and/or absence of clinical signs indicative of ORS after a median follow-up period of 22.5 months confirmed treatment efficacy in all dogs. CONCLUSION: Laparoscopic surgery for ORS in dogs is effective with minimal complications and short convalescence and can successfully replace the conventional, more invasive open surgical procedure.

The use of anti-Müllerian hormone as diagnostic for gonadectomy status in dogs.

In the veterinary practice, there is a need for a diagnostic tool to check the gonadal status in female dogs because it may be difficult to determine whether a female animal has been spayed or whether there are ovarian remnants. Although less prevalent, a similar situation pertains to male dogs. Anti-Müllerian hormone (AMH) is an important regulator of gonadal function and is a specific gonadal product that can be determined in circulation. The objective of this study was to develop and test a canine blood AMH assay as a diagnostic tool to determine the presence of functional gonadal tissue in dogs. A prospective study with a training-validation set paradigm was used. A canine AMH assay was developed and serum and plasma AMH concentrations were determined in blood samples from 46 intact female dogs, 48 spayed females, 50 intact males, and 48 castrated males collected at two separate institutes. Using a training-validation set paradigm, it was found that using cutoff values of 1.1 ng/mL (female) and 5.5 ng/mL (male) AMH, the assay reported excellent specificity and sensitivity of 100% and 90% in female dogs, and good specificity and sensitivity of 100% and 76%, in male dogs, respectively. The sensitivity in male dogs could be further enhanced by including a serum testosterone determination. This newly developed canine AMH assay is a valuable diagnostic tool to determine gonadal status in veterinary medicine.

Identification of a novel kisspeptin with high gonadotrophin stimulatory activity in the dog.

Kisspeptin (KISS1) and its receptor (KISS1r) are essential for normal reproductive function in many species, but the role of kiss1/kiss1r signalling in the dog has not yet been elucidated. The aims of this study were to identify the canine kiss1 and kiss1r genes and to determine gonadotrophin and oestradiol stimulatory activity of KP-10, the shortest biologically active form of KISS1. Canine kiss1 and kiss1r genes were localized by comparing the reference dog genome with relevant human cDNA sequences, using BLASTn software. The amino acid sequence of canine KP-10 (YNWN V FGLR Y ) differs at two positions from human KP-10 (YNWN S FGLR F ). A single bolus of canine KP-10 was administered intravenously to anoestrous Beagle bitches in dosages of 0, 0.1, 0.2, 0.3, 0.5, 1, 5, 10, and 30 µg/kg. Blood samples were collected before and after canine KP-10 administration for the measurement of plasma luteinizing hormone (LH, all doses), follicle-stimulating hormone (FSH) and oestradiol (1-30 µg/kg). From 0.2 µg/kg onwards, canine KP-10 resulted in a rapid and robust rise in plasma LH concentration (max. at 10 min). KP-10 also resulted in a rapid and robust rise in plasma FSH concentration (max. at 10-20 min). Plasma oestradiol concentration increased significantly after dosages of 1, 5, and 10 µg/kg and reached a maximum at 60-90 min. In conclusion, canine KP-10 is a potent kisspeptin which elicits robust gonadotrophin and oestradiol responses in anoestrous bitches, suggesting that canine kiss1/kiss1r are cogent targets for modulating reproduction in dogs.

A distinct expression pattern in mammalian testes indicates a conserved role for NANOG in spermatogenesis.

BACKGROUND: NANOG is a key player in pluripotency and its expression is restricted to pluripotent cells of the inner cell mass, the epiblast and to primordial germ cells. Spermatogenesis is closely associated with pluripotency, because through this process highly specialized sperm cells are produced that contribute to the formation of totipotent zygotes. Nevertheless, it is unknown if NANOG plays a role in this process. METHODOLOGY/PRINCIPAL FINDINGS: In the current study, NANOG expression was examined in testes of various mammals, including mouse and human. Nanog mRNA and NANOG protein were detected by RT-PCR, immunohistochemistry, and western blotting. Furthermore, eGFP expression was detected in the testis of a transgenic Nanog eGFP-reporter mouse. Surprisingly, although NANOG expression has previously been associated with undifferentiated cells with stem cell potential, expression in the testis was observed in pachytene spermatocytes and in the first steps of haploid germ cell maturation (spermiogenesis). Weak expression in type A spermatogonia was also observed. CONCLUSIONS: The findings of the current study strongly suggest a conserved role for NANOG in meiotic and post-meiotic stages of male germ cell development.