A PCR-based NGS protocol for whole genome sequencing of West Nile virus lineage 2 directly from biological specimens.

Lineage 2 West Nile virus (WNV) strain has been involved in a severe outbreak of encephalitis in humans and equines which are in Europe. WNV molecular characterization is important for the development of diagnostic tests, and to obtain molecular information, which is necessary for epidemiological investigations in areas at risk of virus transmission. For whole-genome sequencing of strains of WNV lineage 2, directly from biological specimens, PCR-based NGS protocol developed.

This method is applied to the WNV-positive specimens were obtained from animals, human and mosquito hosts in Greece. Outcomes of the application shows that, even in the case of a low viral titers, developed PCR-based NGS approach capable of providing whole genome sequence of strain lineage 2 WNV.

This study evaluated the performance of biological denitrification in anoxic biofilm sequencing batch reactor (ASBBR) and nitrous oxide (N2O) emissions. After 90 days of operation, chemical oxygen demand of waste and the amount of nitrogen elimination of high-efficiency 94.8% and 95.0%, respectively.

Both polysaccharides and the protein content is reduced in the bound EPS (TB-EPS) and loosely bound EPS (LB-EPS) after the formation of biofilms. According to a typical cycle, the rate of release of N2O associated with the concentration of free nitric acid (FNA) with a maximum value of 3.88 mg / min and the total conversion rate of 1.27%. Two components identified from the model EEM-PARAFAC in soluble microbial products (SMP).

Proteins such as substance for one component changed significantly in the process of denitrification, while humic and fulvic acid-like-like substance for the second component remained relatively stable. Results of high throughput sequencing showed that Lysobacter, Tolumonas and Thauera is the dominant genera, showed the co-existence of autotrophic and heterotrophic denitrifiers in ASBBR.

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