Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Soil microbial community response to land-management and depth,related to the degradation of organic matter in English wetlands: Implications for the in situ preservation of archaeological remains

Wetlands are important habitats not only for their unique ecological value but also because they contain organic material that is fundamental to our understanding of precedent landscape and human past. This study compares the effects of two different land-management regimes on metabolic diversity and bacterial community structure with depth in order to relate them to the process of organic matter degradation and the potential for preservation in situ of organic archaeological artefacts in wetland soils. Soil cores were collected at five depths down to 100 cm from two wetlands sites in England. Environmental variables were monitored and the metabolic capabilities of the microbial community were studied using Biolog Ecoplates^®. DNA was extracted from soil, and the bacterial community structure was examined by polymerase chain reaction followed by denaturing gradient gel electrophoresis (PCR-DGGE). To determine compositional changes in the bacterial community with depth, information about specific groups of bacteria at the site with higher water table (Hatfield Moor) was obtained by cloning and sequencing of 16S rRNA genes. Biolog and DGGE analyses showed depth variation and between-site variation. Carbon substrate utilization and bacterial diversity decreased with increasing depth. The wetland soil under an arable regime in which the water levels were kept elevated, showed higher metabolic capability and bacterial richness when compared with the soil under pasture and subjected to long-standing drainage. Cloning and sequencing showed that Proteobacteria and Acidobacteria were the predominant taxa within the soil profile, but there was a clear shift in bacterial community composition with increasing depth as several taxonomic groups (δ-Proteobacteria and Spirochaetes) were only detectable at 50 cm depth. Because the site with a high and stable water table presented higher metabolic activity and bacterial diversity, it may be that saturated conditions and a high water table are not sufficient to guarantee the preservation in situ of organic material such as archaeological artefacts.