International Journal of Microbiology and Immunology Research Vol. 6(1), pp. 001-007, October 2018 ISSN 2327-7769 ©2018 Academe Research Journals

Full Length Research Paper

Isolation and identification of pathogenic Acanthamoeba using morphological method combined with PCR, in recreational water bodies in the State of San Luis Potosí, Mexico

Ricardo Ortíz Ortega¹, Alejandro Monsalvo Reyes², A. Bernabé Lorenzana Fong¹ and Alonso Vilches-Flores³

¹Proyecto Cy MA, Laboratorio de Microbiología Ambiental, UIICSE, FES IZTACALA, UNAM. Av. de los Barrios, No. 1, Los Reyes Iztacala, Tlalnepantla, Estado de México, México, C.P. 54090.

²Laboratorio de Análisis y Química de ADN UNAM. Av. De los Barrios No. 1, Col. Los, Reyes Iztacala, Tlalnepantla, Estado de México. C.P. 54090.

³UBIMED, FES IZTACALA, UNAM. Av. de los Barrios, No. 1, Los Reyes, Iztacala, Tlalnepantla, Estado de México, México, C.P. 54090.

*Corresponding author. E-mail: diagbouga.serge@gmail.com.

Accepted 14 January, 2017

Abstract

The free-living amoebae (FLA) can be found in any type of environment, but specifically the genus Acanthamoeba is associated with several infections as granulomatous amoebic encephalitis and keratitis amoebic, typical related to the use of contact lenses. To determinate the presences of FLA in a specific media or in infected individuals require at least one week identifying them. In this investigation we used the traditional method of taxonomic identification, combined with the PCR technique to determinate the presence of pathogenic FLA of the genus Acanthamoeba in aquatic environments located in the Huasteca Potosina. Four independent samplings in nine sites were accomplished throughout an annual cycle in The Huasteca zone of the State of San Luis Potosi, including springs, waterfalls, lagoons, and pools fed with thermal waters. 74 collected samples, and, by duplicate were processed by the conventional laboratory methods to detect organizations belonging to the genus Acanthamoeba and analyzed by the technique of PCR, using two primers, one for the detection of Acanthamoeba genus, and the second for the detection of pathogenic strain. In seven selected places, 49 isolations of the genus Acanthamoeba were obtained by the traditional method, most of them were from the recreational center ponds fed with thermal water. The study of pathogenicity in mice showed 31 cases. PCR analysis with primer Ac6 revealed 33 pathogenic samples, the same result obtained by the conventional method. The use of primer GP (P2) confirmed 41 samples with the presence of Acanthamoeba genus previously identified, demonstrating that PCR technique is a rapid and reliable alternative to detect FLA. Their presence in popular resorts and touristic natural water bodies is an alert of a potential health care risk.

Key words: Free-living Acanthamoeba, morphological identification, PCR, recreational water bodies.