The objective of the present study was to describe the proteins from the seminal plasma of buffalo and correlate these proteins with sperm motility. Ejaculates from sixteen Murrah buffalo were used. Semen collection was performed by electroejaculation, and the ejaculate was evaluated by macroscopic (volume) and microscopic analysis (subjective motility and vigor, as well as sperm concentration). After the analysis, the samples were centrifuged (800g for 10 min and 10,000 for 30 min at 4 °C), and the supernatant (seminal plasma) was used to determine total protein concentration by the Bradford method. Based on total protein concentration, an aliquot (50 μg) was taken to conduct protein in-solution digestion for nano-LC-ESI-Q-TOF mass spectrometry analysis. Samples were divided into two groups, minimal (little sperm motility) and greater (typical sperm motility), based on non-hierarchical clustering considering motility and emPAI protein value. The data were analyzed by multivariate statistical analysis using principal component analysis (PCA) and partial analysis of minimum squares discrimination (PLS-DA). Forty-eight proteins were detected in the seminal plasma, and fifteen were common to two groups. There were six proteins that were significantly different between the groups. The main functions of proteins in seminal plasma were catalytic and binding activity. Spermadhesin protein, ribonuclease, 14-3-3 protein zeta/delta and acrosin inhibitor were in greater amounts in seminal plasma from the group with greater sperm motility; prosaposin and peptide YY were in greater amounts in the group with little sperm motility. The proteins detected in the greater motility group were correlated with sperm protection, including protection against oxidative stress, lipid peroxidation, protease inhibition and prevention of premature capacitation and acrosome reaction. In the group with little sperm motility, one of the identified proteins is considered to be an antifertility factor, whereas the function of other identified protein is not definitive. Results from the present study add to the knowledge base about the molecular processes related with sperm motility, and these findings can be used for determining potential markers of semen quality.
The aim of this study was to investigate the effects of different concentrations of oral supplementation with selenium (Se) upon ram sperm parameters. Thirty rams managed in stall under intensive system were used and divided into five groups (six animals per group) as follows: control group (G1) mineral mixture supplementation without Se, group 2 (G2) mineral mixture supplemented with 5 mg/kg Se, group 3 (G3) supplemented with 10 mg/kg Se, group 4 (G4) supplemented with 15 mg/kg Se and group 5 (G5) supplemented with 20 mg/kg Se. For each group, there was an adjustment period of 14 days. The experimental period was 350 days. Every 56 days, the animals were weighed and semen samples were collected by electroejaculation. Semen analysis included volume, mass moviment, total motility, vigour, concentration and morphology. For plasmatic and acrosomal membrane integrity evaluation and mitochondrial membrane potential were used a combination of fluorescent probes. Differences between means values obtained by analysis of variance were verified by Tukey test with 5% probability. There was no statistical difference between treatment groups in relation to volume, mass moviment, total motility, vigour, concentration, plasma and acrosomal membrane integrity (p > .05). Sperm morphology was different between treatment groups, the G1 (0 mg of selenium) had the highest percentage of major defects (11.11 ± 1.11 ; p < .05). It was concluded that selenium decrease the percentage of sperm defects and did not directly influence on ram sperm volume, mass moviment, total motility, vigour, concentration and membrane integrity.
The objective of this study was to evaluate variations in the orbital area, muzzle and vulva surface temperatures and progesterone (P4) concentrations during follicular and luteal phases in Murrah buffalo and whether these temperatures are influenced by the weather patterns. Forty cows were submitted to P4-based hormonal protocol. After P4 device withdrawal transrectal ultrasonography and infrared digital thermography were performed daily until day 16 and on days 20, 24, 28 and 32 to follow the ovulation as well as the vulva, orbital area and muzzle temperatures. In addition, the weather variables were evaluated, as well as rectal temperature (RT) and P4 and cortisol concentration. Vulva, muzzle and orbital area temperatures correlated positively with RT and with weather data. Greater temperatures of the vulva, orbital area and muzzle were detected during the period of estrus. The vulvar surface temperature (VST) was not influenced to a great extent by weather factors during the morning, so this period was chosen to evaluate the influence of the phase of the estrous cycle on VST. The VST was less during days 16, 20, 24 and 28 (diestrus) and P4 concentration was inversely proportional to the VST. Muzzle, orbital area and RT, however, were not of the same pattern. Negative correlations were observed between VST and P4 concentrations. It is concluded that VST undergoes changes during the reproductive phases, correlating with P4 concentration. The weather factors influence the temperatures of the body surface areas, and the morning is the most desirable time to perform the thermographies.
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