M. B. R. Alves

Dinâmica da celularidade do colostro de vacas da raça Holandesa no pós-parto imediato

Avaliou-se a celularidade do colostro de 53 vacas da raca Holandesa, utilizando-se 171 amostras, colhidas apos a primeira e a segunda ordenha pos-parto. Para a analise citologica quantitativa e qualitativa, foram utilizadas as tecnicas de microscopia direta e citocentrifugacao. O total de leucocitos obtido por contagem microscopica direta foi de 0,878x106 e 1,260x106, antes da primeira e da segunda ordenha, respectivamente, e os valores da mediana dos fagocitos obtidos antes da primeira e da segunda ordenha foram de 0,657 e 0,828x106 leucocitos mononucleares, e 0,178 e 0,392x106 leucocitos polimorfonucleares (P<0,0001). Os valores da mediana dos tipos leucocitarios, antes da primeira e da segunda ordenha, foram: 0,640x106 e 0,772 monocitos/celulas epiteliais; 0,136x106 e 0,098 linfocitos; 0,045x106 e 0,203x106 neutrofilos, respectivamente. Concluiu-se que o colostro bovino da segunda ordenha apresentou aumento da quantidade de celulas somaticas e leucocitos polimorfonucleares/mL de colostro. Essas variacoes podem estar relacionadas a importância do colostro aos neonatos, ou a modificacoes fisiologicas e de defesa da glândula mamaria, durante o periodo de adaptacao dela.

An Efficient Technique to Detect Sperm Reactive Oxygen Species: The CellRox Deep Red ® Fluorescent Probe

Heat stress and testicular traumas increase sperm Reactive Oxygen Species (ROS) resulting in Oxidative Stress (OS) and injury of sperm. The fluorescent probe CellROX Deep Red® is not already used to detect ROS in spermatozoa. On this way, this study aims to evaluate its efficiency in detecting ROS in ram sperm. For that, it was used ejaculates from rams performing the in vitro induction of sperm OS in T0, T50 and T100. T0 was semen sample that was not submitted to OS induction, T50 was 50% induced to OS induction and T100 was entire submitted to OS induction. The production of ROS was detected by CellROX Deep Red®. Data polynomial regression analysis was performed and the result of determination coefficient was 0.728. Besides, sixteen rams were submitted to scrotal insulation during 72 hours performing the in vivo induction of OS sperm. Analyses of sperm motility, morphology, DNA fragmentation and ROS (detected by CellROX Deep Red®) were done two times before the scrotal insulation and two times afterwards. Data analysis of variance was performed from the periods (before x after) and it is observed that after scrotal insulation the total and progressive motility decrease (p<0.05) and total and major sperm defects and sperm DNA fragmentation increase (p<0.05). Moreover, after insulation, it was observed increase (p<0.05) in sperm showing moderate and intense OS. Thus, it is possible to conclude that CellROX® fluorescent probe is able to detect ROS production in ram sperm by in vitro and in vivo induction being a new technique to detect sperm OS.

Validation of the CellRox Deep Red® fluorescent probe to oxidative stress assessment in equine spermatozoa

Considering the importance of ROS influence on sperm functionality and some limitations in sperm oxidative stress assessment methods, a field to studies of new techniques are still open. In this sense, the aim of this study is to validate the ROS detection technique through the CellRox Deep Red Reagent® probe in stallion sperm. Four stallions were used and the analyses were conducted on four replicates of semen samples from each of stallion (n = 16). The results of the polynomial regression presented a quadratic effect, high determination coefficient value (R2 = 0.88) and high significant P value (P < 0.0001). The CellRox Deep Red® fluorescent probe is able to detect reactive oxygen species in equine sperm, indicating accurately the occurrence of oxidative stress in stallion semen.

Melatonin Added to Cryopreservation Extenders Improves the Mitochondrial Membrane Potential of Postthawed Equine Sperm

&NA; Reactive oxygen species (ROS) can induce sperm damage, especially after cryopreservation. Melatonin (MEL) is a potent amphiphilic antioxidant, allowing free penetration to any compartment of cell. Ferulic acid (FA) is a phenolic compound exhibiting a wide range of beneficial effects against several pathologies due to its antioxidant property. This study aimed to evaluate the effect of MEL and FA on sperm quality of cryopreserved equine semen. Five ejaculates from four stallions were collected. For each ejaculate, the control group (conventional freezing extender BotuCrio; Botupharma, Botucatu, SP, Brazil) and two different concentrations of each antioxidant (MEL 2 mM, MEL 1 &mgr;M, FA 0.5 mM, and FA 1.2 mM) were tested. Sperm kinetics were assessed by CASA system (SCA). Integrity of plasma and acrosomal membranes and mitochondrial potential were assessed using fluorescent probes PI, Hoechst 33342, FITC‐PSA, and JC‐1. Sperm ROS production was evaluated by CellRox Deep Red. Comparisons among treatments were analyzed by generalized linear model (PROC GLM in SAS, version 9.3), and differences were compared with Duncans test (a value of P ≤ .05 was considered significant). MEL 1 &mgr;M treatment demonstrated higher percentage of sperm cells presenting intact plasma membrane, intact acrosome, and high mitochondrial membrane potential, which was due to higher percentage of sperm cells with high mitochondrial membrane potential. No differences among treatments were observed on ROS evaluation. It was concluded that semen treatment with 1 &mgr;M MEL improves mitochondrial function of equine sperm cells submitted to cryopreservation process. HighlightsMelatonin (MEL) had a better effect on cryopreserved equine sperm when compared with control and ferulic acid treatments.MEL improves sperm membrane integrity in the equine sperm cryopreservation process.MEL showed a direct effect on sperm mitochondria.

Effect of simple and low-cost enrichment items on behavioral, clinical, and productive variables of caged laying hens

ABSTRACT Housing layers in battery cages is a practice still used by many countries but it has been criticized because of its influence on behavioral repertoire of birds. We investigated whether simple and affordable enrichment devices alone impact behavior, foot condition and performance of laying hens housed in conventional cages. Hens were divided into plain cages (CON), cages with perches (PER), and cages with tassels and scratch-pads (ENR), and parameters were evaluated before and after enrichment placement. After perch placement inactivity, drinking and competition for space reduced 35.6%, 40.8% and 70.3%, respectively, whereas social interaction increased 19.3%. Both modifications decreased locomotion (75.0% and 42.4% for PER and ENR respectively) and abnormal behaviors (62.5% and 43.9.4% for PER and ENR respectively). None of the performance variables were affected by ENR or PER. Thermography was more efficient than visual inspection in detecting subclinical bumblefoot, and it confirmed that PER reduced subclinical and clinical cases. Our findings indicate that perches increased welfare-related behaviors and foot health of hens, supporting the use of these inexpensive and highly adaptable alternatives for the enrichment of battery cages.

Prolapso vaginal e uterino em ovelhas

O presente trabalho objetivou realizar um estudo retrospectivo sobre os prolapsos vaginal e uterino em ovelhas atendidas no Servico de Clinica de Bovinos e Pequenos Ruminantes (CBPR) da FMVZ/USP no periodo compreendido entre 2000 a 2010, no qual, foram atendidas 56 ovinos com problemas inerentes ao sistema reprodutivo, dessas, 25 apresentaram prolapso vaginal ou uterino (44,6%). O prolapso vaginal total foi o de maior frequencia (72%). As ovelhas acometidas, em sua maioria, possuiam idade superior a quatro anos (64%), eram sem raca definida (44%) ou da raca Ile de France (40%). As manifestacoes clinicas observadas durante a maioria dos atendimentos foram: taquipneia, taquicardia, mucosas oculares avermelhadas indicando estado de toxemia, decubito esternal ou lateral, apatia e anorexia. O tratamento instituido para todos os casos foi a limpeza, desinfeccao e reintroducao do orgao prolapsado. A sutura de Buhner foi feita em 84% dos casos e a histeropexia em um caso (4%). A evolucao foi satisfatoria em 80% dos casos atendidos, nos demais casos (20%) observou-se obito da femea acometida. Do total de obitos, os prolapsos vaginais foram responsaveis por 60% (3/5) e os prolapsos uterinos por 40% (2/5). A etiologia dos prolapsos nao foi definida nos casos atendidos, sendo esses associados com o periodo pos-parto em sua maioria (56%), provavelmente associados com quadros de hipocalcemia, altas concentracoes sericas de estrogeno e hipertonia uterina. Alem disso, a predisposicao genetica nao pode ser descartada.