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Benign Prostatic Hyperplasia (BPH) is a pathological condition that directly interferes with the reproductive potential of senile dogs, by leading to prostate enlargement and sperm injury, which in turn may compromise sperm freezeability. Moreover, albeit finasteride treatment reduces prostatic volume and blood supply and maintains seminal quality and testicular integrity, the effects of sperm samples submitted to cryopreservation after the finasteride treatment are still unknown. Thus, the aim of this study was to evaluate frozen-thawed semen of BPH dogs, as well as dogs subjected to BPH pharmacological treatment with finasteride. For such purpose, 20 dogs were previously selected and assigned to three experimental groups, according to BPH diagnosis and treatment with finasteride: Control (n = 9), BPH Group (n = 5) and BPH-Finasteride Group (n = 6). Semen was subjected to one-step cryopreservation protocol with tris-fructose-citric acid extender with 5% glycerol and thawed at 37°C for 30 sec. Fresh and post-thaw sperm samples were evaluated for macroscopic parameters, sperm concentration, sperm motility kinetics, sperm mitochondrial activity and potential, oxidative stress, plasmatic and acrosome membrane integrity, sperm DNA fragmentation and sperm binding test on perivitelic membrane of chicken egg yolk. Regarding fresh semen, BPH-Finasteride group had the lowest ejaculate visual aspect (opacity), higher frequency of sperm flagellar beating (BCF) and percentage of sperm with medium velocity. Control group had the highest percentage of sperm DNA integrity compared to BPH group. For the frozen-thawed semen, Control group presented the highest percentage of spermatozoa with high mitochondrial activity. However, the BPH-Finasteride group showed higher number of sperm bound to the perivitelline membrane of chicken egg yolk compared to the BPH Group. Conversely, BPH group had higher percentage of DNA damage. In conclusion, the ejaculate of BPH dogs has higher susceptibility to cryoinjury, whereas finasteride-treated dogs have increased spermatozoa functional performance, suggesting a promising use of BPH dogs as semen donors in sperm cryopreservation programs.

Effects of conservative treatment on uterine blood flow and morphometric findings are still unknown in bitches. Thus, this study aimed to compare uterine changes of pyometra bitches subjected to distinct modes of treatment. Pyometra bitches were assigned to: OHE (ovariohysterectomy immediately after diagnosis), Aglepristone (days 1, 2 and 8) and Associative (aglepristone treatment coupled with cloprostenol for 7 days) groups. After 9 days, bitches were ovariohysterectomized. Before surgery, uterine area was measured ultrasonographically and the uterine artery Doppler velocimetry analyzed blood flow velocity and indexes. Uterine horns were classified according to resistance index (RI) as more compromised and less compromised. Endometrial vasculature was quantitatively evaluated by color flow Doppler. Blood samples were collected to determine nitric oxide (NO) concentrations. Histological uterine structures were quantified by stereology and VEGF-A (vascular endothelial growth factor) and eNOS were (endothelial nitric oxide synthase) immunohistochemically analyzed. Aglepristone and Associative groups had lower uterine area and vascularization, and higher blood flow velocity and indexes compared to OHE group. Less compromised horn of Associative group had higher blood flow velocity compared to OHE group. Aglepristone group presented lower inflammatory infiltrate and larger uterine stroma. Associative group had lower volume density and absolute surface of endometrial cysts and lower VEGF-A expression for glandular epithelium and stromal cells. Blood NO and e-NOS immunostaining were not different among groups. In conclusion, association between aglepristone and prostaglandin is more effective in decreasing uterine vascularization and modulating uterine blood flow. Moreover, associative therapy promotes marked morphological changes.

Cystic endometrial hyperplasia–pyometra complex is a canine endometrial disorder, considered the most common and important among uterine diseases. The treatment of choice is ovariohysterectomy, but medical treatments have become an alternative. However, no studies have been performed in order to evaluate uterine hemodynamic changes during medical treatment for pyometra bitches. Therefore, the aim of this study was to compare the effectiveness of two medical protocols by means of clinical (heart rate, respiratory rate and body temperature), medullar response (complete blood count (CBC)) and uterine hemodynamic evaluation in pyometra bitches. For such purpose, 10 bitches with pyometra were assigned to two groups: Aglepristone Group (n = 5) and Associative Group (aglepristone + prostaglandin; n = 5). The Associative Group had altered respiratory rate and decreased body temperature. The Aglepristone Group had decreased plasma protein, however, leukocyte count reduced over time for both therapeutic protocols. Uterine area and vascularization score decreased throughout treatment. There was a significant reduction in uterine perimeter, area and vascularization score. The Associative Group had lower final diastolic velocity and higher systole:diastole ratio (S/D), pulsatility index (PI) and resistance index (RI). In conclusion, both medical treatments are effective in reversing clinical and CBC changes of pyometra, especially aglepristone, however they cannot fully restore local uterine changes in a short-term evaluation. Conversely, associative therapy was more effective in decreasing uterine vascularization and modulating uterine blood flow.

During fertilization, spermatozoa interact with the zona pellucida (ZP) through the binding between the acrosome and proteins 2 and 3 (ZP2 and ZP3). The perivitelline membrane of chicken egg yolk is homologous to the mammalian ZP3, which allows the binding of sperm of several species. The aim of this study was to standardize and evaluate the efficiency of sperm-binding to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation. For this purpose, nine post-thaw sperm samples were used, which were divided into two aliquots: the first was kept in water bath at 37ºC (live sample) and the second was submitted to cold shock to induce cellular damage (dead sample). The two aliquots were mixed on five proportions, corresponding to 0%, 25%, 50%, 75%, and 100% of viable cells, and the binding test was performed by analyzing the number of spermatozoa bonded to the perivitelline membrane by means of computerized assessment of sperm motility (CASA) or conventional microscopy. Additionally, samples were submitted to sperm motility analysis, evaluation of plasmatic and acrosomal membrane integrity, and sperm mitochondrial activity. The sperm-binding test to the perivitelline membrane of chicken egg yolk was considered a feasible sperm analysis test for both fertilizing capacity and overall sperm attributes evaluation, mainly when the analysis is performed by a conventional microscope, which expands its practicality to the majority of canine reproduction laboratories.