Two distinct introductions of the West Nile virus in Portugal disclosed by phylogenetic analysis of genomic sequences.

In this report, we describe genomic sequencing and analysis of different West Nile virus strains isolated from mosquitoes in the south of Portugal (Alentejo and Algarve) at two different time points (1971 and 2004, respectively). Phylogenetic analysis indicated different origins for the two recorded introductions of WNV in our country, with strains segregating in different sub-clades within lineage 1a. PTRoxo (isolated in 1971) was found to be very similar to an Egyptian WNV strain isolated in 1951, while the viruses isolated in 2004 formed a statistically well-supported group with WNV strains isolated over the last decade in countries lining the Mediterranean (Morocco, Italy, and France). Analyses of the putative amino acid sequences showed all the main expected features described for viral mature proteins except for the absence of the N-glycosylation site in the envelope glycoprotein of PTRoxo, which may be reflected as attenuated neurovirulence and neuroinvasive phenotypes.