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The Mediterranean subtype probably originated in the Mediterranean area approximately 20?years ago [16]. It is conceivable that the 2011 WNV genomes detected in Veneto originated in the Mediterranean area. Less likely, their origin could be in other unknown areas where these types of viruses could be circulating without notice and be brought to the Mediterranean by unknown mechanisms. The two genome sequences might have been introduced independently in the Veneto region or, alternatively, they could have arisen as the two sequences evolved and diverged locally from a common ancestor already present in Veneto or in neighbouring regions, in a situation of endemic transmission in rural cycles remaining silent, as recently suggested for other Mediterranean WNV strains [7]. However, the presence of significant amino acid changes http://www.selleckchem.com/products/Neratinib(HKI-272).html in the viral polyprotein of the two genomes, as shown in Fig. S1 (see Supporting information), argues against this hypothesis and indicates two lines of independent evolution that separated a long time ago, unless recombination events had occurred and generated a rapid evolution. In particular, the viral polyproteins had amino acid changes unique to these genomes that were not previously RVX-208 reported in other WNV isolates as well as other amino acid changes that are not generally found in Eastern European and Mediterranean isolates, as detailed in the Supporting information (Fig. S1). In September 2011, in the same area where the novel human WNV lineage 1a sequences were detected (Fig.?1), a dead wild collared dove was found to be positive for WNV lineage 2, with sequence similarity to the Hungary/04 strain, and the entomological surveillance led to the identification of mosquito pools positive for both WNV lineage 1 and 2 [17]. The presence of different WNV strains in a relatively small geographical area is a novel finding with unpredictable impact on human disease, which requires enhanced surveillance and further investigation. The results of this study were presented at the 22nd European Congress of Clinical Microbiology and Infectious Diseases, London, UK, BMS-777607 mw 31 March to 3 April 2012 (abstracts S378 and O465). This research was funded by the Veneto Region and by the EU FP7 project WINGS (grant no. 261426) to Giorgio Pal��. The authors declare that they have no conflict of interest. Figure S1. Summary of amino acid changes detected in the polyprotein of West Nile virus (WNV) full genome sequences compared with the consensus sequence of Eastern European and Mediterranean WNV subtypes, as described by May et al. [May et al. J Virol 2011]. The WNV strains are included in the figure have the following accession numbers: Marocco 96-111-horse (AY701412); France PaAn001-horse (AY268132); Marocco WNV04.