Uso de dimetil-formamida associada ou não ao glicerol na criopreservação de sêmen caprino Use of dimethylformamide associated or not with glycerol for goat semen cryopreservation

O objetivo neste trabalho foi avaliar os efeitos da dimetil-formamida na criopreservação de sêmen caprino, por meio de testes in vitro. As partidas de sêmen foram congeladas com os diluidores leite desnatado-gema, associado a diferentes crioprotetores e concentrações: 7% de glicerol (T1); 3,5% de glicerol e 3,5% de dimetil-formamida (T2) e 5% de dimetil-formamida (T3). O sêmen foi centrifugado e envasado em palhetas de 0,5 mL, sendo resfriado por 40 minutos, atingindo 5,0°C e permanecendo nesta temperatura por mais 1 hora e 20 minutos. Os parâmetros avaliados in vitro foram a motilidade progressiva e o vigor espermático, a integridade acrossômica, a integridade da membrana plasmática (HO) e a reação acrossômica. Observou-se perda de 30,0% da motilidade inicial quando as amostras foram submetidas aos procedimentos de criopreservação. A perda de integridade da membrana plasmática, avaliada pelo teste hiposmótico (HO) foi de 19%. As lesões de acrossoma aumentaram em 3% durante o teste de termo-resistência (TTR) lento. Não houve diferença entre os tratamentos quanto aos aspectos de motilidade nos tempos de 0,00; 0,05; e 1,00 horas do TTR e vigor em todos os tempos. No tempo de 2,00 horas do TTR, registrou-se diferença entre os tratamentos 1 e 2, de modo que a motilidade no tratamento 2 foi superior às demais. Os valores de integridade de membrana plasmática, integridade acrossômica e reação acrossômica pós-descongelamento não diferiram entre os tratamentos, indicando que a dimetil-formamida não foi superior ao glicerol na manutenção da qualidade espermática após a criopreservação.<br>The objective of this trial was to evaluate the efficiency of dimethylformamide on cryopreservation of goat semen using semen physical analyses and complementary tests. Alpine and Saanen bucks from the Dairy Goat Experimental Station- DZO were used. Semen samples were frozen with a yolk-skimmed milk diluent and cryoprotectors with different concentrations as follows: 7% glycerol (T1), 3.5% glycerol plus 3.5% dimethylformamide (T2) or 5% dimethylformamide (T3). Semen was centrifuged, diluted, and bottled in 0.5 mL straws followed by cooling for 40 minutes until reach 5.0°C and kept at this temperature for additional 80 minutes. Samples were then exposed to liquid nitrogen vapor for 15 minutes and finally frozen. Thawing was done in a waterbath at 37°C for 50 seconds. The following variables were evaluated in vitro: sperm progressive motility and vigor, acrosomal integrity and reaction, and plasmatic membrane integrity. The cryopreservation procedures reduced 30.0% of the initial motility and 19% of plasmatic membrane integrity that was evaluated using the hyposmotic swelling test (HOST). It was observed an increaseof 3% in acrosomal lesions when samples were submitted to a slow thermoresistance test (TRT). No significant differences in sperm motility were observed among treatments at 0, 5, and 60 minutes of TRT as well as in sperm vigor at all TRT times. However, at 120 minutes of TRT a significant difference in sperm motility was found comparing T1 and T2 while T3 was intermediate. There were no significant differences on plasmatic membrane integrity, acrosomal integrity, and post-thawing acrossomal reaction across treatments. It can be concluded that dimethylformamide can be a viable alternative for goat semen cryopreservation used alone or in association with glycerol

Semen cryopreservation protocols of Mangalarga Marchador stallions

The effect of the utilization of three semen protocols (Inra 82®, Merck Gema and Botu-crio®) and two filling techniques (0.25 and 0.50 mL straws) in Mangalarga Marchador stallions were studied in this experiment. Sperm parameters were assessed during processing and post-freezing. No interactions between the protocols and type of filling were observed, so they were assessed separately. Sperm parameters were not altered when the extender was added to the centrifugation; however, there was reduction of motility and strength when freezing extenders were added. The Botu-crio® protocol preserved the parameters of total and progressive sperm motility, smoothed path velocity (µm/s), straight line velocity (µm/s), track velocity (µm/s) and the average and fast spermatozoa percentage better than the others. No difference between the extenders for the percentage of sperm integrity was observed. There was no difference in the responses studied on the filling techniques. The stallions presented better freezing with the use of the Botu-crio® protocol. The best post-freezing viability results were found for semen frozen using the Botu-crio® protocol and there were no differences concerning the sperm quality comparing 0.25 and 0.50 mL straws

Giants of the Amazon: How does environmental variation drive the diversity patterns of large trees?

For more than three decades, major efforts in sampling and analyzing tree diversity in South America have focused almost exclusively on trees with stems of at least 10 and 2.5 cm diameter, showing highest species diversity in the wetter western and northern Amazon forests. By contrast, little attention has been paid to patterns and drivers of diversity in the largest canopy and emergent trees, which is surprising given these have dominant ecological functions. Here, we use a machine learning approach to quantify the importance of environmental factors and apply it to generate spatial predictions of the species diversity of all trees (dbh ≥ 10 cm) and for very large trees (dbh ≥ 70 cm) using data from 243 forest plots (108,450 trees and 2832 species) distributed across different forest types and biogeographic regions of the Brazilian Amazon. The diversity of large trees and of all trees was significantly associated with three environmental factors, but in contrasting ways across regions and forest types. Environmental variables associated with disturbances, for example, the lightning flash rate and wind speed, as well as the fraction of photosynthetically active radiation, tend to govern the diversity of large trees. Upland rainforests in the Guiana Shield and Roraima regions had a high diversity of large trees. By contrast, variables associated with resources tend to govern tree diversity in general. Places such as the province of Imeri and the northern portion of the province of Madeira stand out for their high diversity of species in general. Climatic and topographic stability and functional adaptation mechanisms promote ideal conditions for species diversity. Finally, we mapped general patterns of tree species diversity in the Brazilian Amazon, which differ substantially depending on size class