RESUMO
Wild birds in the Orders Anseriformes and Charadriiformes are the natural reservoir for avian influenza (AI) viruses. Transmission within these aquatic bird populations occurs through an indirect fecal-oral route involving contaminated water on shared aquatic habitats. In order to better understand the influence that aquatic environments exert on AI transmission and maintenance in the wild-bird reservoir system, we determined the duration of persistence for 12 wild-bird origin AI viruses under natural ranges of pH, salinity, and temperature. Viral persistence was measured using a laboratory-based distilled water model system. The AI viruses varied in their response to each of the examined variables, but, generally, the viruses were most stable at a slightly basic pH (7.4-8.2), low temperatures (<17 degrees C), and fresh to brackish salinities (0-20,000 parts per million (ppm)). Alternatively, the AI viruses had a much shorter duration of persistence in acidic conditions (pH<6.6), warmer temperatures (>32 degrees C), and high salinity (>25,000 ppm). The results of this research suggest that the pH, temperature, and salinity in natural aquatic habitats can influence the ability of AI viruses to remain infective within these environments. Furthermore, these results provide insight into chemical and physical properties of water that could enhance or restrict AI virus transmission on an aquatic bird habitat.
Assuntos
Anseriformes/virologia , Charadriiformes/virologia , Reservatórios de Doenças/veterinária , Influenza Aviária/virologia , Orthomyxoviridae/patogenicidade , Microbiologia da Água , Animais , Linhagem Celular , Reservatórios de Doenças/virologia , Cães , Concentração de Íons de Hidrogênio , Influenza Aviária/transmissão , Salinidade , TemperaturaRESUMO
BACKGROUND: Effective influenza surveillance requires new methods capable of rapid and inexpensive genomic analysis of evolving viral species for pandemic preparedness, to understand the evolution of circulating viral species, and for vaccine strain selection. We have developed one such approach based on previously described broad-range reverse transcription PCR/electrospray ionization mass spectrometry (RT-PCR/ESI-MS) technology. METHODS AND PRINCIPAL FINDINGS: Analysis of base compositions of RT-PCR amplicons from influenza core gene segments (PB1, PB2, PA, M, NS, NP) are used to provide sub-species identification and infer influenza virus H and N subtypes. Using this approach, we detected and correctly identified 92 mammalian and avian influenza isolates, representing 30 different H and N types, including 29 avian H5N1 isolates. Further, direct analysis of 656 human clinical respiratory specimens collected over a seven-year period (1999-2006) showed correct identification of the viral species and subtypes with >97% sensitivity and specificity. Base composition derived clusters inferred from this analysis showed 100% concordance to previously established clades. Ongoing surveillance of samples from the recent influenza virus seasons (2005-2006) showed evidence for emergence and establishment of new genotypes of circulating H3N2 strains worldwide. Mixed viral quasispecies were found in approximately 1% of these recent samples providing a view into viral evolution. CONCLUSION/SIGNIFICANCE: Thus, rapid RT-PCR/ESI-MS analysis can be used to simultaneously identify all species of influenza viruses with clade-level resolution, identify mixed viral populations and monitor global spread and emergence of novel viral genotypes. This high-throughput method promises to become an integral component of influenza surveillance.