Development of qPCR platform with probes for quantifying prevalent and biomedically relevant human gut microbial taxa.

Nowadays the advent of innovative high-throughput sequencing allows obtaining high-quality microbiome profiling. However, PCR-based tests are still considered the "golden standard" for many clinical applications. Here, we designed a qPCR-based platform with fluorescent-labeled oligonucleotide probes for assessing human gut microbiome composition. The system allows conducting qualitative and semiquantitative analysis for 12 prokaryotic taxa that are prevalent in the human gut and associated with diseases, diet, age and other factors. The platform was validated by comparing microbiome profile data obtained with two different methods - the platform and high-throughput 16S rRNA sequencing - across 42 stool samples. The test can form the basis for precise and cost-efficient microbiome assay for large-scale surveys including clinical trials with interventions related to diet and disease risks.

Why Do Testate Amoeba Optima Related to Water Table Depth Vary?

This study focusses on the ecology of testate amoeba species in peatlands of the southern taiga of Western Siberia. To estimate the influence of the trophic state of mires on species optima related to water table depth, a separate study of three calibration datasets including ombrotrophic, minerotrophic and the combined habitats was conducted. In the datasets obtained separately from ombrotrophic and minerotrophic mires, the water table depth was the main factor affecting testate amoeba assemblages. However, the trophic state (specifically pH and ash content) was more important factor in the combined dataset, including all of the studied mires. For 36 testate amoeba species, which were found in the ombrotrophic and minerotrophic mire habitats, their species optima, obtained separately in ombrotrophic and minerotrophic datasets, differed significantly from each other. Some of these species preferred minerotrophic conditions, while others preferred ombrotrophic ones. For all species, the trophic state of the mires affected the values of the species optima related to water table depth, as revealed in the form of a threshold effect. In extreme conditions, the species were more sensitive to the trophic status than to the water table depth, and their optimum related to water table depth was distorted. Variation of the optimum was observed in those species that inhabited both ombrotrophic and minerotrophic mires due to the fact that mires with a different trophic status were included in the training sets. The optima did not vary for species inhabiting only ombrotrophic or only minerotrophic mires.