Original Paper

Parasitology Research

, Volume 111, Issue 1, pp 383-392

First online:

Molecular characterization of Acanthamoeba isolated in water treatment plants and comparison with clinical isolates

  • A. MagnetAffiliated withParasitology Laboratory, San Pablo CEU University, Urbanización Montepríncipe
  • , A. L. GalvánAffiliated withParasitology Laboratory, San Pablo CEU University, Urbanización MontepríncipeGrupo de Parasitología. Escuela de Microbiología, Universidad de Antioquia
  • , S. FenoyAffiliated withParasitology Laboratory, San Pablo CEU University, Urbanización Montepríncipe
  • , F. IzquierdoAffiliated withParasitology Laboratory, San Pablo CEU University, Urbanización Montepríncipe
  • , C. RuedaAffiliated withParasitology Laboratory, San Pablo CEU University, Urbanización Montepríncipe
  • , C. Fernandez VadilloAffiliated withParasitology Laboratory, San Pablo CEU University, Urbanización Montepríncipe
  • , J. Pérez-IrezábalAffiliated withHospital Cruces
  • , K. BandyopadhyayAffiliated withCenter for Global Health, Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention
  • , G. S. VisvesvaraAffiliated withNational Center for Emerging and Zoonotic Infectious Diseases, Division of Foodborne, Waterborne and Environmental Diseases, Centers for Disease Control and Prevention
    • , A. J. da SilvaAffiliated withCenter for Global Health, Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention
    • , C. del AguilaAffiliated withParasitology Laboratory, San Pablo CEU University, Urbanización MontepríncipeFacultad de Farmacia, Urbanización Montepríncipe Email author 

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Abstract

A total of 116 samples (44 clinical specimens and 72 environmental samples) have been analyzed for the presence of Acanthamoeba. The environmental samples (ESs) were collected from four drinking water treatment plants (DWTP, n = 32), seven wastewater treatment plants (n = 28), and six locations of influence (n = 12) on four river basins from the central area of Spain (winter–spring 2008). Water samples were concentrated by using the IDEXX Filta-Max® system. Acanthamoeba was identified in 65 of the 72 ESs by culture isolation (90.3%) and 63 by real-time PCR (87.5%), resulting in all sampling points (100%) positive for Acanthamoeba when considering both techniques and all the time period analyzed. Nine of the 44 clinical specimens were positive for Acanthamoeba. Seventeen Acanthamoeba strains (eight from four DWTP and nine from clinical samples) were also established in axenic-PYG medium. Twenty-four of the ESs and the 17 Acanthamoeba sp. strains were genotyped as T4/1, T4/8, and T4/9. The eight strains isolated from the DWTP samples were inoculated in nude mouse to ascertain their potential pathogenicity in this model. Animals that were inoculated died or showed central nervous system symptoms 9 days post-inoculation. Examination of immunofluorescence-stained brain and lung tissue sections showed multiple organisms invading both tissues, and re-isolation of throphozoites was successful in these tissues of all infected animals. For the first time, potentially pathogenic Acanthamoeba T4 has been detected in 100% of different types of water samples including tap water and sewage effluents in the central area of Spain suggesting a potential health threat for humans especially for the contact lens wearers.