Prolactin,Androstenedione and IGF1 Serum Concentrations During Induced Follicular Growth by eCG Administration in the Bitch

The oestrus cycle in the domestic bitch, a monoestrous species, differs considerably from that of other veterinary domestic animals species. In the bitch the combined use of eCG and hCG is effective to induce oestrus predictably and safely (Stornelli et al., Theriogenology, 78, 2012 and 1056). Although several studies were done to describe the hormonal changes during the canine oestrus cycle, to our knowledge none was done to describe the hormonal changes during induced follicular growth after the administration of eCG. The aim of this work was to study prolactin (PRL), insulin‐like growth factor (IGF1) and androstenedione (ANDR) serum concentrations during follicular growth induced by a single dose of eCG administered to late anoestrous bitches. PRL and ANDR concentrations were lower before than after eCG TRT (before eCG vs pro‐oestrus, oestrus and dioestrus; 4.3 ± 1.8 ng/ml vs 6.5 ± 1.6 ng/ml, p < 0.05; 0.08 ± 0.2 ng/ml vs 0.42 ± 0.16 ng/ml, p < 0.05). Conversely, IGF1 concentrations were similar before and after eCG TRT (286.0 ng/ml ±32.2, p > 0.53). Additionally, PRL concentrations were similar before oestrus compared to during oestrus and dioestrus (6.9 ± 1.7 ng/ml, p > 0.19). Furthermore, IGF1 concentrations were higher before and during oestrus compared to first day of dioestrus (286.1 ± 29.8vs 200.4 ± 29.2 ng/ml, p < 0.01). On the contrary, ANDR concentrations were lower before and during oestrus compared to first day of diestrum (0.35 ± 0.17 ng/ml and 0.38 ± 0.15 vs 0.68 ± 0.17 ng/ml, p < 0.05). These results show that treatment with a single injection of 50 IU/kg of eCG in late anoestrous bitches successfully induced changes in follicular growth which were paralleled with changes in PRL, IGF1 and ANDR serum concentration similar to those occurring during a normally occurring oestrous cycle. In addition, our results suggest that IGF1 in the bitch could play an important role in ovarian folliculogenesis.     

Expression of Occludin in Canine Testis and Epididymis

Tight junctions (TJ) in inter-Sertoli junctional areas and epididymal epithelia are important for the formation of blood-testis barrier (BTB) and blood-epididymal barrier (BEB). In this study, the expression of occludin, an integral member of the TJ, was verified in canine testis and epididymis. Both low molecular weight (MW) (25-28 kDa) forms as well as high MW (68-72 kDa) forms of occludin were detected in the testis and epididymis using Western blot. The relative amount of the high MW forms of occludin vs low MW forms was higher in the testis than in the epididymis. Some difference in the composition of different MW forms of occludin was found along the segments of epididymis, suggesting the possible correlation between cellular composition of occludin proteins and paracellular permeability of epithelia along the epididymal tubule. In the testis, intense occludin immunoreactivity was found in the basally located inter-Sertoli junctional area. Diffused immunoreactivity of occludin was also found in the cytoplasm of Sertoli cells. A similar pattern of zonula occludens-1 immunoreactivity was found in the cytoplasm of Sertoli cells, suggesting that occludin was not confined to the inter-Sertoli junctional areas and that subcellular localization of occludin in the Sertoli cells was dynamically regulated during spermatogenesis in canine testis. In the epididymis weak immunoreactivity was found in the apical sides and cytoplasm of epithelial cells.     

Effects of Mating on Plasma Concentrations of Testosterone, Cortisol, Oestrone Sulphate and 15-Ketodihydro-PGF2α in Stallions

Very little information is available regarding the physiological mechanisms involved in the normal sexual activity in the stallion and, in particular, the endocrine control of reproduction is still not clearly understood. This experiment was designed to determine the short-term effect of sexual stimulation on plasma concentrations of testosterone, cortisol, oestrone sulphate and 15-ketodihydro-PGF(2alpha) in stallions. Semen samples were collected from 10 lighthorse stallions of proven fertility using a Missouri model artificial vagina. At the same time, blood samples were collected from the jugular vein with heparinized tubes, 20 and 10 min before oestrous mare exposure, at exposure and 10, 20, 30 min after dismounting. Testosterone concentrations showed a sharp rise 10 min after mating (p < 0.001), reached a plateau, and then showed a further increase 30 min after mating (p < 0.001). Cortisol concentrations increased 10 min after mating (p < 0.001) and remained at high levels in the subsequent samples taken. A peak of oestrone sulphate was observed 10 min after mating (p < 0.001). 15-Ketodihydro-PGF(2alpha) concentrations decreased rapidly at the moment of the exposure of the stallions to an oestrous mare (p < 0.05), returned to pre-mating concentrations and then decreased again 30 min after mating (p < 0.05).     

17β‐Estradiol,Progesterone and Testosterone Concentrations in Cystic Fluids and Response to GnRH Treatment after Emptying of Ovarian Cysts in Dairy Cows

The aim of this study was to determine possible links between steroidogenic activity of single ovarian cysts and response to intramuscular treatment with 20 μg of buserelin (GnRH‐analogue) after cyst emptying, in pluriparous Friesian cows bearing a singleton cyst treated not earlier than 55 days post‐partum. Progesterone, 17β‐estradiol and testosterone were determined in cystic fluids collected by needle aspiration of the cyst. Of the cows, 75.6% began ovarian cyclicity within 30 days after treatment with a conception rate of 64.7%. In this study it was found that as progesterone concentration in cystic fluids rose, the number of positive responses to the treatment fell.     

Involvement of Protein cAMP‐dependent Kinase,Phospholipase A2 and Phospholipase C in Sperm Acrosome Reaction of Chinchilla lanigera

The mechanisms involved in fertilization are the centre of attention in order to determine the conditions required to reproduce in vitro the events that take place in vivo, with special interest in endangered species. Previous data from mouse sperm, where acrosome reaction (AR) occurs more often in the interstitium of the cumulus oophorus, contribute to strengthen the use of progesterone as a physiological inducer of this process. We studied the participation of protein kinase A (PKA), phospholipases A₂ and C (PLA₂, PLC) in the AR induced by progesterone from Chinchilla epididymal spermatozoa. The addition of db‐cAMP to the incubation medium caused an increase of 58% in the AR, while the use of H89 (30 μm ), a PKA inhibitor, reflected a decrease of 40% in the percentage of reacted gametes. The assays conducted with arachidonic acid showed a maximum increase of 23% in the AR. When gametes were pre‐incubated with PLA₂ inhibitors, a dose‐dependent inhibitory effect was observed. The addition of phorbol12‐myristate13‐acetate (10 μm ) revealed higher percentages of AR induction (60%). When PLC was inhibited with neomycin and U73122, a dose‐dependent decrease in AR percentages was observed. Combined inhibition of PKA, PLA₂ and PLC, AR values similar to control were obtained. This work shows evidence, for the first time in Chinchilla, that progesterone activates the AC/cAMP/PKA system as well as sperm phospholipases and that these signalling pathways participate jointly and cooperatively in AR. These results contribute to the understanding of the complex regulation that is triggered in sperm after the effect of progesterone.     

Seasonal Changes in Testicular Cell Morphology in Domestic Male Cats (Felis catus)

The aim of this study was to asses the variation in the morphology of the seminal epithelium in relation to natural photoperiod in male cats. Tom cats (n = 240) were castrated every other week throughout the year. Each testis was fixed in Bouin's solution and cut into sections. The percentage of tubules with round spermatids (RS), elongated spermatids (ES), tailed spermatids (TS), mature spermatids (MS) and the number of Sertoli cells (SC) and Leydig cells (LC) were recorded in each sample. Testicles from males during short days (SHD) had a higher percentage of tubules with RS and ES compared to testicles from males during long days (LHD, 31.3 ± 0.6 vs 2.1 ± 0.6%, p < 0.001; 30.9 ± 0.7 vs 11.0 ± 0.7%, p < 0.001). Conversely, testicles from males during SHD had a lower percentage of tubules with TS and MS compared to testicles from males during LHD (24.5 ± 0.8 vs 29.7 ± 0.8%, p < 0.01; 13.1 ± 1.2 vs 57.0 ± 1.2%, p < 0.01). Furthermore, testicles from males during SHD had a higher number of SC and lower number of LC compared to testicles from males during LHD (11.4 ± 0.1 vs 8.0 ± 0.1%, p < 0.01; 19.2 ± 1.0 vs 38.0 ± 1.0%, p < 0.01). In conclusion, there are seasonal changes in testis cell morphology in the tom which may be related to seasonal sperm production.     

Acute febrile diarrhoea in horses: 86 cases (1986–1991)

SUMMARY: Eighty-six horses presented for examination at the Rural Veterinary Centre between January 1986 to December 1991 with acute diarrhoea. The average age of affected horses was 3.2 ± 0.2 years (mean ± SE), with 69% three years or younger. Sixty-one horses were male (36 stallions, 25 geldings) and 83 horses were Thoroughbreds (47) or Standardbreds (36). Sixty-six horses were undergoing routine training at the time of onset of diarrhoea. Eight horses were afflicted with a non-specific illness within one to five days before the onset of diarrhoea, whereas eight horses developed diarrhoea during or within 48 h of discontinuation of antimicrobial therapy. Three horses developed the diarrhoea within 24 h of road transportation. Clinically, the disorder was characterised by a fever, sudden onset of profuse diarrhoea, clinical evidence of dehydration (estimated to be 5 to 12% of body weight at the time of admission) and shock. Degenerative leucopaenia, hyponatraemia, hypochloraemia, hyperkalaemia, hyperglycaemia and azotaemia were characteristic laboratory findings. Laminitis was a sequel in about 30% of cases. The cause of diarrhoea remained undetermined in most cases. Salmonellas were isolated from faecal or tissue samples in only two cases. Similarly, there was no evidence of seroconversion to Erhlichia risticii in 17 cases. Sixty-two of the horses survived the acute phase of the disease in response to supportive care. In horses that did not survive, necropsies were performed and revealed sanguineous or turbid peritoneal fluid. The colonic and caecal walls were oedematous and thickened with serosal congestion and discolouration of these organs evident grossly. Histopathological examination of the caecum and colon revealed diffuse congestion, oedema, inflammatory reactions and submucosal vascular thrombosis with varying degrees of mucosal devitalisation. Most cases presented in this series represent a distinct syndrome of diarrhoea characterised by acute diarrhoea predominantly affecting young horses in training with concomitant signs of fever and toxaemia.