Comparative study of PCR-sexing procedures using bovine embryos fertilized with sex-sorted spermatozoa

Sex determination in bovine embryos is a useful tool in reproductive biotechnology. This work compares two techniques of embryo sexing by polymerase chain reaction (PCR. Embryos were produced in vitro with sex-sorted sperm and two techniques of DNA lysis were tested (proteinase K versus heat shock). Subsequently, halves of each lysed sample was amplified both by amelogenin and BRY4a/SAT1 primers. Male embryos treated by both digestion methods and amplified by BRY4a/SAT1 gave higher rates of false negatives. Amelogenin amplification failed with embryos previously digested by proteinase K. In contrast, the lysis method allowed obtaining the best accuracy in terms of sex verification when using amelogenin amplification

Environmental determinants of macroinvertebrate diversity in small water bodies: insights from tank-bromeliads

The interlocking leaves of tank-forming bromeliads (Bromeliaceae) collect rainwater and detritus, thus creating a freshwater habitat for specialized organisms. Their abundance and the possibility of quantifying communities with accuracy give us unparalleled insight into how changes in local to regional environments influence community diversity in small water bodies. We sampled 365 bromeliads (365 invertebrate communities) along a southeastern to northwestern range in French Guiana. Geographic locality determined the species pool for bromeliad invertebrates, and local environments determined the abundance patterns through the selection of traits that are best adapted to the bromeliad habitats. Patterns in community structure mostly emerged from patterns of predator species occurrence and abundance across local-regional environments, while the set of detritivores remained constant. Water volume had a strong positive correlation with invertebrate diversity, making it a biologically relevant measure of the pools' carrying capacity. The significant effects of incoming detritus and incident light show that changes in local environments (e.g., the conversion of forest to cropping systems) strongly influence freshwater communities. Because changes in local environments do not affect detritivores and predators equally, one may expect functional shifts as sets of invertebrates with particular traits are replaced or complemented by other sets with different traits

Algal blooms increase heterotrophy at the base of boreal lake food webs-Evidence from fatty acid biomarkers

Physical defenses and grazer avoidance of the bloom-forming microalga Gonyostomum semen may reduce the direct coupling between phytoplankton and higher trophic levels and result in an increased importance of alternative basal food resources such as bacteria and heterotrophic protozoans. To assess the importance of algal and heterotrophic food resources for zooplankton during G. semen blooms and the effects of zooplankton diets on a higher consumer, we analyzed the fatty acid composition of zooplankton and the invertebrate predator Chaoborus flavicans from eight lakes along a gradient in the predominance of G. semen relative to other algae and the duration of G. semen blooms. The proportion of fatty acids of bacterial origin increased significantly along the G. semen gradient in all consumers studied. In addition, the proportion of polyunsaturated fatty acids (PUFA) decreased in cladocerans. These results suggest that heterotrophic pathways can compensate for a reduced trophic coupling between phytoplankton and filter-feeding zooplankton. The lower PUFA content in cladoceran prey from lakes at the higher end of the G. semen gradient did not affect the PUFA content of the predator C. flavicans, suggesting selective assimilation and retention of PUFA and/or feeding on other, more PUFA-rich prey

Estudio comparativo de dos métodos de sexaje por PCR en embriones bovinos producidos con semen sexado

Sex determination in bovine embryos is a useful tool in reproductive biotechnology. This work compares two techniques of embryo sexing by polymerase chain reaction (PCR). Embryos were produced in vitro with sex-sorted sperm and two techniques of DNA lysis were tested (proteinase K versus heat shock). Subsequently, halves of each lysed sample was amplified both by amelogenin and BRY4a/SAT1 primers. Male embryos treated by both digestion methods and amplified by BRY4a/SAT1 gave higher rates of false negatives. Amelogenin amplification failed with embryos previously digested by proteinase K. In contrast, the lysis method allowed obtaining the best accuracy in terms of sex verification when using amelogenin amplification.La determinación del sexo en embriones bovinos es una herramienta útil en las biotecnologías reproductivas. Este trabajo compara dos técnicas de determinación del sexo en embriones mediante la reacción en cadena de la polimerasa (PCR). Los embriones fueron producidos in vitro con semen sexado y se analizaron dos técnicas de lisis de ADN (proteinasa K y choque térmico). Posteriormente, la mitad de cada muestra lisada fue amplificada por la amelogenina y por los cebadores BRY4a/SAT1. Los embriones machos tratados por ambos métodos de digestión y amplificados por BRY4a/SAT1 dieron mayores tasas de falsos negativos. La amplificación de la amelogenina no dio resultados en embriones previamente digeridos por proteinasa K; sin embargo, el método de lisis por choque térmico junto con la amplificación por amelogenina permitió obtener la mayor precisión en términos de verificación de sexo