Abstract Logistic and economical limitations are often the causes of dog owners not accurately monitoring the estrous cycle and the optimal insemination time. The aim of this study was to evaluate in vivo early embryonic development in bitches, after the analysis of sequential vaginal cytologies associated to single progesterone measurement and single laparoscopic insemination with high quality semen (fresh and with high spermatozoa concentration) or low-quality semen (frozen/thawed and with low spermatozoa concentration) at 48 h post- ovulation time predicted on a single progesterone measurement. Ten bitches were inseminated with 250 x 106 fresh spermatozoa (80% motility), and ten with 80 x 106 frozen/thawed spermatozoa (60% motility) in the cranial part of each uterine horn. Seven days later, ovariohysterectomy was performed and the oviducts and uterine horns and body were flushed to recover embryos and unfertilized oocytes. In 80% of the bitches inseminated with fresh and 50% of bitches inseminated with frozen/thawed semen, embryos at 2 to 8 cells stage were recovered mostly from the, oviducts. This study indicates that pregnancies can be obtained with a single laparoscopic intrauterine insemination after single serum progesterone measurement, although with a low number of embryos. This result should be taken into account in case economic or logistic restrictions that affect the possibility of owners to plan an accurate monitoring of the optimal breeding time using fresh and frozen semen. concentration lowquality frozenthawed thawed 4 post 25 10 (80 motility, motility , 60% 60 (60 horn later oocytes 50 1 (8 6 (6 5 (

Abstract Semen collection methods vary greatly and rely on the practitioner's expertise, available materials, and the specific behavioral traits of the male animals involved. When it comes to domestic cats, wild felids, and canids, semen collection is particularly challenging. Thus, given the difficulty of semen collection in these species, pharmacological semen collection (PSC) stands out since it is a quick and straightforward method that does not require specific equipment. The PSC consists of administering α2-adrenergic receptor agonist drugs, mainly medetomidine, and dexmedetomidine, aiming semen release into the urethra with posterior urethral catheterization and sperm recovery. This technique was primarily described in domestic cats and wild felids, and despite the decreased seminal volume, the retrieved semen is highly concentrated and presents good quality. However, further studies are required to optimize semen collection in domestic dogs and wild canids. Therefore, the purpose of this review is to provide a comprehensive overview of the research developed on pharmacological semen collection (PSC) in the past few decades. The objective is to equip professionals with the essential knowledge required for the efficient application of this technique in both domestic and wild canids and felids and to make a valuable contribution to conservation efforts and the preservation of biodiversity, aligning with the principles of One Conservation. practitioners practitioner s expertise materials involved challenging Thus species (PSC equipment α2adrenergic αadrenergic α2 adrenergic α drugs medetomidine dexmedetomidine recovery volume quality However Therefore decades biodiversity Conservation

Resumo Objetivou-se avaliar a qualidade espermática do sêmen de touros suplementados com selênio (Se) na dieta. Foram utilizados 16 touros Brangus, igualmente distribuídos em grupo controle (GC) e grupo Se (GSe − 0,1mg de Se/kg de MS de dieta). O experimento teve duração de 75 dias, e os animais foram suplementados por 60 dias. Foram realizadas quatro coletas de sêmen durante o período (0, 30, 60 e 75 dias) por animal. As amostras foram avaliadas quanto a motilidade e vigor espermáticos, integridade e funcionalidade da membrana plasmática (teste de expansão hiposmótico - HIPO) e viabilidade espermática e reação acrossomal (coloração tripla - TRI). Após avaliação, estas foram diluídas em meio Tris-gema com 5% de glicerol, envasadas (40x106 espermatozoides/palheta), resfriadas, congeladas e armazenadas em nitrogênio líquido até a análise. Após descongelação, foram submetidas às mesmas avaliações descritas para o sêmen fresco. Não houve interferência da suplementação com Se nas variáveis vigor espermático, HIPO e TRI do sêmen fresco e descongelado. Porém, constatou-se queda na motilidade espermática do GSe comparativamente ao GC no sêmen fresco (P=0,0035) e descongelado (P=0,0067) após 60 dias de suplementação. Portanto, a suplementação de Se na dieta não foi efetiva na promoção de melhorias dos parâmetros espermáticos de touros Brangus.

Abstract The aim of the present study was to evaluate sperm quality of bulls supplemented with selenium (Se) in the diet. Sixteen Brangus bulls were randomly divided in two groups: control (GC) and Se (GSe - 0.1mg Se/kg dietary DM). The experiment lasted 75 days and the animals were supplemented for 60 days. Four semen collections (0, 30, 60 and 75 days) per animal, were performed during the experimental period. Sperm motility and vigor, plasma membrane integrity and functionality (hyposmotic swelling test - HOST) and sperm viability and acrosome reaction (triple staining -TRI) were assessed. After immediate analysis, samples were diluted in Tris-egg yolk extender with 5% glycerol, packed (40x106 spermatozoa / straw), chilled, frozen and stored in liquid nitrogen until analysis. After thawing, sperm motility and vigor, HOST and TRI were performed. No significance was noticed on sperm vigor, HOST and TRI of fresh and post-thawed semen after dietary selenium supplementation. However, sperm motility decreased in GSe compared to GC in fresh (P = 0.0035) and post- thawed (P = 0.0067) semen samples after 60 days of supplementation. Therefore, dietary selenium supplementation was ineffective to improve semen parameters of fresh and post-thawed semen of Brangus bulls.

A fisiopatologia da hiperplasia prostática benigna (HPB) não está totalmente compreendida, no entanto, a diidrotestosterona é o principal hormônio envolvido. Recentemente, o efeito da toxina botulínica A (TB-A) foi investigado, mostrando que esta induz atrofia do parênquima e redução do volume prostático. Com base nisso, este estudo teve como objetivos comparar os efeitos da administração da TB-A com a orquiectomia no tratamento da HPB, além de avaliar os efeitos da TB-A sobre a libido e qualidade do sêmen. Para tanto, 16 cães adultos foram submetidos à castração ou administração de 500U de TB-A, e avaliados durante 16 semanas. A orquiectomia mostrou-se um excelente tratamento, promovendo redução de 80% do volume prostático. Aplicação da TB-A não ocasionou alterações na libido, ereção ou qualidade e características seminais. Por outro lado, reduziu significamente o volume da próstata. Os resultados sugerem que a administração intraprostática de TB-A é um tratamento alternativo efetivo e sem efeitos colaterais para cães destinados a programas de reprodução. No entanto, esta terapia apresenta reduções inferiores do volume prostático, quando comparada com a orquiectomia.

The pathogenesis of benign prostatic hyperplasia (BPH) is not completely known, however, dihydrotestosterone is a hormone involved in enlargement of the prostate. Recently, the effect of botulinum toxin A (BT-A) was investigated and has shown to induce atrophy of the gland. Based on that, the aims of this study were to compare the effects of administration of BT-A with orchiectomy in the treatment of BPH, in addition to assessing the effects of BT-A on libido and semen quality. Therefore, 16 sexually intact male dogs were submitted to orchiectomy or administration of 500U BT-A, and evaluated for 16 weeks. Orchiectomy presented excellent results, reducing the prostate volume up to 80%. Administration of BT-A did not interfered on libido, erection or semen characteristics. On the other hand, reduced significantly the prostate volume. The results suggest that intraprostatic injection of BT-A is an effective alternative treatment without adverse effects for valuable breeding dogs with BPH. However, this therapy showed lower reduction of prostate volume when compared to orchiectomy.

O objetivo do presente estudo foi determinar a ação de diferentes concentrações de toxina botulínica tipo A (TB-A) sobre os parâmetros seminais, perfis bioquímicos e proteicos do plasma seminal de cães com hiperplasia prostática benigna (HPB). Dezoito cães hígidos, não orquiectomizados com HPB foram divididos em três grupos, os quais foram submetidos à injeção intra-prostática de solução salina (grupo controle - GC), 250UI (GI) ou 500UI (GII) de TB-A. Amostras seminais foram coletadas previamente aos tratamentos e após 2, 4 e 8 semanas. Os parâmetros seminais assim como os valores de pH e concentrações de proteínas totais (TP), cloretos totais (CT), cálcio (Ca), potássio (K), sódio (Na) do plasma seminal foram mensurados após as coletas. O perfil proteico do fluido prostático foi estabelecido por meio de eletroforese SDS-PAGE. Não foram constatadas diferenças significativas quanto aos parâmetros espermáticos e perfil bioquímico do plasma seminal intragrupos e intergrupos (P>0,05). À SDS-PAGE foram identificadas 31 bandas proteicas com pesos moleculares de 3,9 a 106,2kDA, em todos os tratamentos e durante todo o período de avaliação. Dessa forma, concluiu-se que, independentemente da dose utilizada, a injeção intra-prostática de TB-A não altera os parâmetros seminais, assim como os perfis bioquímico e proteico do plasma seminal de cães com HPB.

This study aimed to determine the effects of different concentrations of botulinum toxin type A (BT-A) on semen parameters, and seminal plasma biochemical and protein profiles of dogs with benign prostatic hyperplasia (BPH). Eighteen sexually intact male dogs with BPH were randomly divided in three groups, and received an intraprostatic injection of saline solution (control group - CG), 250UI (GI) or 500UI (GII) of BT-A under transabdominal ultrasound guidance. Semen was collected at baseline, 2, 4 and 8 weeks after treatment. Semen parameters were determined and seminal plasma pH, total protein (TP), total chlorides (TC), calcium (Ca), potassium (K), and sodium (Na) concentrations were assessed. One-dimensional sodium dodecyl sulfatepolyacrilamide gel eletrophoresis (SDS- PAGE) was performed to determine seminal plasma protein profile. Sperm parameters and seminal plasma pH, TP, TC, Ca and K mean values did not change significantly at any time point and among treated groups (P>0.05). The SDS-PAGE analysis of the pooled fractions identified 31 protein bands with molecular weights ranging from 3.9 to 106.2kDA in all treatment groups during the entire evaluation period. Regardless the used dose, intraprostatic BT-A injection do not alter semen parameters and seminal plasma biochemical and protein profiles of dogs with BPH.