The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Fluoroquinolones (FQs) are synthetic antimicrobials commonly used in intensive poultry farming to treat chronic respiratory disease, colibacillosis, and fowl cholera. As these drugs are also important in treating infections in humans, their use in poultry has been restricted in some countries to avoid the development of antimicrobial-resistant bacteria. In this work, a novel simple and efficient LC-MS/MS method using solid phase extraction (SPE), ESI ionization and multiple reaction mode (MRM) has been developed and validated to determine the two most common FQ residues: enrofloxacin (ENR) and ciprofloxacin (CIP) in chicken target tissues (muscle, liver, kidney, fat and skin) and in plasma. The method is shown to be sensitive, with limits of quantification (LOQ) between 1 ng g À1 and 5 ng g À1 , recoveries 93-115% and correlation coefficients greater than 0.99. This novel methodology allows faster and simpler sample workflow compared to previously reported methodologies, making it ideal for high-throughput screening and confirmation of both ENR and CIP meeting the international regulation levels.

The effectiveness of induction of the acrosome reaction (AR) test as a parameter to in vitro estimate embryo production (IVP) in Nelore breed and the AR pattern by the Trypan Blue/Giemsa (TB) stain were evaluated. Frozen semen samples from ten Nelore bulls were submitted to AR induction and were also evaluated for cleavage and blastocyst rates. The treatments utilized for AR induction were: control (TALP medium), TH (TALP medium + 10μg heparin), TL (TALP medium + 100μg lysophosphatidylcholine) and THL (TALP medium + 10μg heparin + 100μg lysophosphatidylcholine). Sperm acrosomal status and viability were evaluated by TB staining at 0 and after 4h incubation at 38°C. The results obtained for AR presented a significant difference (P<0.05) in the percentage of acrosome reacted live sperm after 4h of incubation in the treatments that received heparin. The cleavage and blastocyst rates were 60% and 38% respectively and a significant difference was observed among bulls (P<0.05). It was founded a satisfactory model to estimate the cleavage and blastocyst rates by AR induction test. Therefore, it can be concluded that the induction of the AR test is a valuable tool to predict the IVP in Nelore breed.Keywords: cattle, fertility, semen, acrosome reaction, fertilization Palavras-chave: bovino, fertilidade, sêmen, reação acrossomal, fertilização RESUMO Avaliou-se a eficiência da técnica de indução da reação acrossomal (RA) como parâmetro para estimar a produção in vitro (PIV) de embriões Nelore e analisou-se o padrão de RA pela técnica de coloração Azul de Tripan/Giemsa (TB). Amostras de sêmen congelado de dez touros foram submetidas à indução da RA e avaliadas quanto a taxa de clivagem e blastocisto. Os tratamentos utilizados para indução da RA foram: controle (meio TALP), TH (meio TALP + 10μg heparina), TL (meio TALP + 100μg lisofosfatidilcolina) e THL (meio TALP + 10μg heparina + 100μg lisofosfatidilcolina

This study aimed to assess the capacity of Morada Nova (MN) and Santa Inês (SIN) rams to maintain body and testicular homeothermy under thermal challenge. For 5 days in the summer, 16 males (SIN = 7 and MN = 9) underwent a heat tolerance test, i.e., period 1-animals maintained in the shade (11 to 12 h); period 2-animals exposed to sunlight (12 to 13 h); and period 3-animals returned to the shade (13 to 14 h). The respiratory rate, heart rate, rectal temperature, and infrared surface temperatures (IRT) of the trunk, back, eyeball, and testicles were assessed in each period. The index of capacity of tolerance to insolation (ICTI), which indicates the animals' level of adaptability, was calculated for each animal. Semen quality and testicular parenchyma integrity were assessed before and after the thermal challenge. Statistical analyses were performed at 5% significance. In period 1, the variables had baseline values for both genotypes. In period 2, the variables involved in thermolysis significantly increased (P< 0.05), which matches a thermal discomfort situation. In period 3, the variables returned to baseline values and some values were lower than those in period 1. Semen quality and testicular parenchyma integrity suffered no negative effects with the thermal challenge. IRT ocular and IRT testicular were positively correlated (P < 0.05). It is concluded that MN and SIN rams had efficient thermolytic mechanisms that favor preserving gonadal functionality. The animals were considered resilient to a thermal challenge. In addition, infrared thermography was an efficient tool to verify body and testicular thermoregulation.