Population and Genetic Study of Vibrio cholerae from the Amazon Environment Confirms that the WASA-1 Prophage Is the Main Marker of the Epidemic Strain that Circulated in the Region

Abstract

Vibrio cholerae is a natural inhabitant of many aquatic environments in the world. Biotypes harboring similar virulence-related gene clusters are the causative agents of epidemic cholera, but the majority of strains are harmless to humans. Since 1971, environmental surveillance for potentially pathogenic V. cholerae has resulted in the isolation of many strains from the Brazilian Amazon aquatic ecosystem. Most of these strains are from the non-O1/non-O139 serogroups (NAGs), but toxigenic O1 strains were isolated during the Latin America cholera epidemic in the region (1991-1996). A collection of environmental V. cholerae strains from the Brazilian Amazon belonging to pre-epidemic (1977-1990), epidemic (1991-1996), and post-epidemic (1996-2007) periods in the region, was analyzed. The presence of genes related to virulence within the species and the genetic relationship among the strains were studied. These variables and the information available concerning the strains were used to build a Bayesian multivariate dependency model to distinguish the importance of each variable in determining the others. Some genes related to the epidemic strains were found in environmental NAGs during and after the epidemic. Significant diversity among the virulence-related gene content was observed among O1 strains isolated from the environment during the epidemic period, but not from clinical isolates, which were analyzed as controls. Despite this diversity, these strains exhibited similar PFGE profiles. PFGE profiles were significant while separating potentially epidemic clones from indigenous strains. No significant correlation with isolation source, place or period was observed. The presence of the WASA-1 prophage significantly correlated with serogroups, PFGE profiles, and the presence of virulence-related genes. This study provides a broad characterization of the environmental V. cholerae population from the Amazon, and also highlights the importance of identifying precisely defined genetic markers such as the WASA-1 prophage for the surveillance of cholera.

Contribuição pioneira do Instituto Evandro Chagas para a saúde ambiental na Amazônia em 25 anos da Seção de Meio Ambiente

“…Com o passar dos anos, esses estudos evoluíram do isolamento, identificação e quantificação de enterobactérias utilizando métodos de cultivo em meios de enriquecimento e aqueles seletivos e indicadores, para pesquisa em nível molecular de diferentes espécies e gêneros de bactérias, inclusive pesquisa de genes de virulência e resistência aos antimicrobianos e metais. Uma importante coleção de isolados bacterianos das famílias Enterobacteriaceae, Vibrionaceae e Aeromonadaceae foi constituída no Laboratório de Microbiologia Ambiental (LMA), de modo que permitiu também a realização de estudos de genética populacional de bactérias, com a identificação de importante marcador molecular entre cepas patogênicas que circularam na Amazônia em epidemia causada por bactéria 34 . Isso possibilitou a elucidação, a nível molecular, dos mecanismos de fermentação da sacarose em cepas amazônidas de Vibrio cholerae O1 da sétima pandemia, que são sacarose negativas, diferindo do padrão de cepas epidêmicas que são sacarose positivas 35 .…”