DNA Status on Thawed Semen from Fighting Bull: A Comparison Between the SCD and the SCSA Tests

The assessment of sperm chromatin status is compulsory in a complete spermiogram. Here we applied the sperm chromatin structure assay (SCSA) and the sperm chromatin dispersion (SCD) test to assess the chromatin status of three fighting bulls. Cryopreserved semen (two straws/bull) were analysed by duplicate after thawing and after 6 h at 37°C with and without oxidative stress (1 m m FE²⁺). Results (SCD: percentage of spermatozoa with halo; SCSA: SD-DFI, %DFI and HDS) were analysed for differences between bulls and treatments, sensitivity and specificity (receiver operating characteristic curves) and repeatability (repeatability coefficients as 2SD of duplicate differences).%DFI for the three bulls was below 2% at 0 h, indicating no risk for fertility according to previous reports. It increased slightly for two of the bulls after FE²⁺ treatment (%DFI < 5%) and more pronouncedly for the other bull (C, %DFI∼10%), which merits further investigation. SCD rendered higher percentage of halos for bull C, but could not discriminate between samples with and without oxidizing treatment (AUC: 0.52). SCSA (%DFI) showed a high discriminating ability between treatments (AUC: 0.96). The repeatability coefficient was also higher for SCD (5.9) than for %DFI (1.8), indicating lower repeatability for SCD. Overall, %DFI might be the most useful parameter for assessing sperm chromatin on fighting bull. SCD might yield different information than SCSA, hence further research is warranted.     

The Impact of Biochar Addition on Nutrient Leaching and Soil Properties from Tillage Soil Amended with Pig Manure

Poses dam in the Seine River estuary acts as receptacle of water drain-offs from highly urbanized and industrialized catchment area; therefore, this water is highly contaminated by trace metals. Most trace elements are mainly bound to particulate matter and are incorporated rapidly into the sediments. Scavenging of these metals in the sediments can be reversible due to several perturbations so as sediments also act as a source of pollutants for the overlying water. For instance, natural events (tide, flood, storm) and anthropogenic processes (water management actions) can cause disturbance of sediments and subsequent remobilization of pollutants to the water column, thereby posing a potential threat for aquatic organisms. The purpose of this study was to evaluate the mobility of trace metals by different methods in the Seine estuary sediments. The surface sediment sampled at Poses dam was characterized by high pollution level of Cd, Cu, Zn, and Pb. The estimation of metal bioavailability through ratio ΣSEM/AVS (simultaneously extracted metals/acid volatile sulfides) indicates a potential bioavailability of trace metals. The chemical partitioning using the European Community of Bureau of Reference sequential extraction method revealed that over 85, 82, and 80 % of the total Cd, Zn, and Pb, respectively, were found to be associated with the exchangeable and reducible fractions of the sediment. Another approach used consists in the quantification of dissolved metals released by sediment resuspension experiments in laboratory under controlled conditions. The results indicated that metals are released rapidly from sediment with a sharp peak at the beginning of the experiment, followed by a fast coprecipitation and/or adsorption processes on the suspended particles. Also, the Cd, Pb, and Ni mobility is higher compared to that of the other metals.     

Effects of Cryopreservation on Bull Spermatozoa Distribution in Morphometrically Distinct Subpopulations

Assisted sperm morphometry analysis (ASMA) was used in this study to determine the effects of cryopreservation on bull spermatozoa distribution in morphometrically distinct subpopulations. Ejaculates were collected from five bulls and were divided. One portion was diluted at 30 degrees C in a skim milk-egg yolk medium, containing glycerol. A microscope slide was prepared from single extended sperm samples prior to freezing. The remainder of each sample was frozen in nitrogen vapours. After thawing, sperm smears were prepared as described above. All slides were air dried and stained with Hemacolor. The sperm-head dimensions for a minimum of 200 sperm heads were analysed from each sample by means of the Sperm-Class Analyser (SCA), and the mean measurements recorded. Our results showed that applying the ASMA technology and multivariate cluster analyses, it was possible to determine that three separate subpopulations of spermatozoa with different morphometric characteristics coexist in bull ejaculates (large, average and small spermatozoa). The mean values of each sperm head dimension among the three subpopulations of spermatozoa were significantly different (p < 0.001). Besides, there were significant (p < 0.001) differences in the distribution of these three sperm subpopulations between fresh and thawed samples. Thus, the percentage of representation of the subpopulation that includes those spermatozoa whose dimensions are the biggest, decreased from 52.06% in extended fresh samples to 15.51% in the thawed ones. Contrarily, the percent of representation of the subpopulation containing the smallest spermatozoa, increased from 8.70% in extended fresh samples to 34.04% in the thawed ones. In conclusion, the present study confirms the heterogeneity of sperm head dimensions in bull semen, heterogeneity that vary through the cryopreservation procedure.