Genotyping of clinical isolates of Acanthamoeba genus in Venezuela

Carolina Wagner; María Reyes-Batlle; María Alejandra Vethencourt Ysea; Mónica V. Galindo Pérez; Carmen Guzmán de Rondón; Anaibeth J. Nessi Paduani; Angelyseb Dorta Pérez; Atteneri López-Arencibia; Ines Sifaoui; María Virginia Pérez de Galindo; Eva Pérez de Suárez; Enrique Martínez-Carretero; Basilio Valladares; José E. Piñero; Jacob Lorenzo-Morales

doi:10.1515/ap-2016-0110

Article

Genotyping of clinical isolates of Acanthamoeba genus in Venezuela

Carolina Wagner , María Reyes-Batlle , María Alejandra Vethencourt Ysea , Mónica V. Galindo Pérez , Carmen Guzmán de Rondón , Anaibeth J. Nessi Paduani , Angelyseb Dorta Pérez , Atteneri López-Arencibia , Ines Sifaoui , María Virginia Pérez de Galindo , Eva Pérez de Suárez , Enrique Martínez-Carretero , Basilio Valladares , José E. Piñero and Jacob Lorenzo-Morales

Published/Copyright: October 24, 2016

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From the journal Acta Parasitologica Volume 61 Issue 4

Abstract

Free-living amoebae of Acanthamoeba genus are opportunistic pathogens distributed worldwide. Strains included in this genus are causative agents of a fatal encephalitis and a sight-threating keratitis in humans and other animals. In this study, 550 clinical samples which were collected between 1984 and 2014 from different patients with suspected infections due to Acanthamoeba were initially screened for the presence of this amoebic genus at the Laboratorio de Amibiasis-Escuela de Bioanálisis at the Universidad Central de Venezuela. Samples were cultured in 2% Non-Nutrient agar plates seeded with a layer of heat killed Escherichia coli. From the 550 clinical samples included in this study, 18 of them were positive for Acanthamoeba genus after culture identification. Moreover, positive samples were confirmed after amplification of the Diagnostic Fragment 3 (DF3) of the Acanthamoeba18S rDNA genus and sequencing was carried out in order to genotype the isolated strains of Acanthamoeba. Furthermore, the pathogenic potential of the strains was checked by performing thermotolerance and osmotolerance assays. Sequencing of the DF3 region resulted in the identification of genotype T4 in all the isolated strains. Moreover, most isolates were thermotolerant or both thermotolerant and osmotolerant and thus were classified as potentially pathogenic strains. To the best of our knowledge, this is the first report on the molecular characterization at the genotype level of Acanthamoeba strains in Venezuela.

Keywords: Acanthamoeba spp; Venezuela; clinical samples; genotype

Acknowledgements

This work was supported by the grants RICET (project no. RD12/0018/0012 of the programme of Redes Temáticas de Investigación Cooperativa, FIS), Spanish Ministry of Health, Madrid, Spain and the Project FIS PI13/00490 "Protozoosis Emergentes por Amebas de Vida Libre: Aislamiento, Caracterización, Nuevas Aproximaciones Terapéuticas y Traslación Clínica de los Resultados" from the Instituto de Salud Carlos III and Project ref. AGUA3 "Amebas de Vida Libre como Marcadores de Calidad del Agua" from CajaCanarias Fundación. CWA was supported by a grant from Consejo de Desarrollo Científico y Humanístico-Universidad Central de Venezuela (Becario Exterior 2014-2015). MRB and ALA were funded by Becas de Investigación Obra Social La Caixa-Fundación CajaCanarias para Posgraduados de la Universidad de La Laguna. Convocatoria 2014. IS was supported by a grant from the Laboratoire Matériaux-Molécules et Applications, IPEST, and Ayudas para estancias estudiantes de posgrado e investigadores del continente americano y africano 2015 from the ULL. JLM was supported by the Ramón y Cajal Subprogramme from the Spanish Ministry of Economy and Competivity RYC-2011-08863.

References

Bermúdez A., Díaz O., Pérez M.V., Pérez E. 1993. Acanthamoeba in the Ophthalmology Service of the University Hospital of Caracas (Acanthamoeba en el Servicio de Oftalmología del Hospital Universitario de Caracas). Anales del Instituto Barraquer (Barcelona), 24, 63-68Search in Google Scholar

Bermúdez A., Pérez de Galindo M.V., Martínez Y., Pérez de Suarez E., Nieto T. 1998. Identification of Acanthamoeba spp. in 3 patients with conjunctivitis (Identificación de Acanthamoeba spp. en 3 pacientes con conjuntivitis). Anales del Instituto Barraquer (Barcelona), 27, 157-163Search in Google Scholar

Booton G.C., Visvesvara G.S., Byers T.J., Kelly D.J., Fuerst P.A. 2005. Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. Journal of Clinical Microbiology, 43, 1689-169310.1128/JCM.43.4.1689-1693.2005Search in Google Scholar PubMed PubMed Central

Cabello-Vílchez A.M., Reyes-Batlle M., Montalbán-Sandoval E., Martín-Navarro C.M., López-Arencibia A., Elias-Letts R., Guerra H., Gotuzzo E., Martínez-Carretero E., Piñero J.E., Maciver S.K., Valladares B., Lorenzo-Morales J. 2014. The isolation of Balamuthia mandrillaris from environmental sources from Peru. Parasitology Research, 113, 2509-251310.1007/s00436-014-3900-2Search in Google Scholar PubMed

Conde C., Machado R. 2010. Molecular characterization of Acanthamoeba isolates from ocular samples at the Bioanalysis School.-UCV using PCR-RFLP of the 18S ribosomal subunit (Caracterización molecular por PCR-RFLP de la subunidad ribosomal 18S, de aislados de Acanthamoeba spp. mantenidos en el laboratorio de Amibiasis de la Escuela de Bioanálisis-UCV provenientes de muestras oculares de pacientes. Trabajo presentado para optar por título de Licenciado en Bioanálisis,) May 2010Search in Google Scholar

Corsaro D., Walochnik J., Köhsler M., Rott M.B. 2015. Acanthamoeba misidentification and multiple labels: redefining genotypes T16, T19, and T20 and proposal for Acanthamoeba micheli sp. nov. (genotype T19). Parasitology Research, 114, 2481-249010.1007/s00436-015-4445-8Search in Google Scholar PubMed

Culbertson C.G., Smith J.W., Cohen H.K., Minner J.R. 1959. Experimental infection of mice and monkeys by Acanthamoeba. American Journal of Pathology, 3, 185-197Search in Google Scholar

Chinchilla M., Castro Elfaro M., Portilla E. 1979. Free-living amoebae as causative agents of meningoencephalitis. First reports in Costa Rica (Amebas de vida libre productoras de meningoencefalitis. Primeros hallazgos en Costa Rica). Revista Latinoamericana de Microbiología, 21, 135-142Search in Google Scholar

Dorta A., Wagner C., Pérez de G. M.V., Blanco de M.J., Galindo M., Nessi A., Vethencourt M.A., Bandes A., Guzmán de Rondón C. 2012. Evaluation of the biological activity of the metanolic and hexanolic extracts of Pera distichophyla against an Acanthamoeba strain isolated from a corneal ulcer (Evaluación de la actividad biológica de los extractos hexanólico (Eh) y metanólico (Em) de Pera distichophylla sobre un aislado de Acanthamoeba spp. proveniente de una úlcera corneal). Revista IberoLatinoamericana de Parasitología, 271, 42-54Search in Google Scholar

García-Tamayo J., González J.E., Martínez A.J. 1980. Primary Amoebic Encepahlitis and Granulomatous Amoebic Encephalitis. Comparative study of two Venezuelan cases. (Meningoencefalitis amibiana primaria y encefalitis granulomatosa amibiana. Estudio comparativo de dos casos venezolanos). Acta Médica Venezolana, 27, 84-91Search in Google Scholar

Gast R.J. 2001. Development of an Acanthamoeba-specific reverse dot-blot and the discovery of a new ribotype. Journal of Eukaryotic Microbiology, 48, 609-61510.1111/j.1550-7408.2001.tb00199.xSearch in Google Scholar PubMed

Gast R.J., Ledee D.R., Fuerst P.A., Byers T.J. 1996. Subgenus systematics of Acanthamoeba: four nuclear 18S rDNA sequence types. Journal of Eukaryotic Microbiology, 43, 498-50410.1111/j.1550-7408.1996.tb04510.xSearch in Google Scholar PubMed

Geisen S., Fiore-Donno A.M., Walochnik J., Bonkowski M. 2014. Acanthamoeba everywhere: high diversity of Acanthamoeba in soils. Parasitology Research, 113, 3151-315810.1007/s00436-014-3976-8Search in Google Scholar PubMed

Hewet M.K., Robinson B.S., Monis P.T., Saint C.P. 2003. Identification of a new Acanthamoeba18S rRNA gene sequence type, corresponding to the species Acanthamoeba jacobsi Sawyer, Nerad and Visvesvara, 1992 (Lobosea: Acanthamoebidae). Acta Protozoologica, 42, 325-329Search in Google Scholar

Horn M., Fritsche T.R., Gautom R.K., Schleifer K.H., Wagner M. 1999. Novel bacterial endosymbionts of Acanthamoeba spp. related to the Paramecium caudatum symbiont Caedibacter caryophilus. Environmental Microbiology, 1, 357-36710.1046/j.1462-2920.1999.00045.xSearch in Google Scholar PubMed

Khan N.A. 2006. Acanthamoeba: biology and increasing importance in human health. FEMS Microbiology Reviews, 30, 564-59510.1111/j.1574-6976.2006.00023.xSearch in Google Scholar PubMed

Khan N.A. 2009. Acanthamoeba: Biology and Pathogenesis. Caister Academic Press: Norfolk, Great Britain, pp. 290Search in Google Scholar

Kong H.H., Chung D.I. 2002. Mitochondrial DNA Restriction Fragment Length Polymorphism (RFLP) and 18S Small-Subunit Ribosomal DNA PCR-RFLP Analyses of Acanthamoeba Isolated from Contact Lens Storage Cases of Residents in Southwestern Korea. Journal of Clinical Microbiology, 40, 1119-120610.1128/JCM.40.4.1199-1206.2002Search in Google Scholar PubMed PubMed Central

Köhsler M, Leitsch D, Fürnkranz U, Duchêne M, Aspöck H, Walochnik J. 2008. Acanthamoeba strains lose their abilities to encyst synchronously upon prolonged axenic culture. Parasitology Research, 1022, 1069-107210.1007/s00436-008-0885-8Search in Google Scholar PubMed

Lorenzo-Morales J., Khan N.A., Walochnik J. 2015. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment. Parasite, 22, 1010.1051/parasite/2015010Search in Google Scholar PubMed PubMed Central

Luongo V., Ugarte-Pelayo A., Galindo M., Vethencourt M.A. 2012. Moelcular characterization of Acanthamoeba spp. isolates (Caracterización molecular de aislados de Acanthamoeba spp.) Acta científica de la Sociedad Venezolana de Bioanalistas, 13-15, 27-33Search in Google Scholar

Ma P., Visvesvara G.S., Martinez A.J., Theodore F.H., Daggett P.M., Sawyer T.K. 1990. Naegleria and Acanthamoeba infections: review. Review Infectious Diseases, 12, 490-51310.1093/clinids/12.3.490Search in Google Scholar PubMed

Magnet A., Henriques-Gil N., Galván-Diaz A.L., Izquierdo F., Fenoy S., del Aguila C. 2014. Novel Acanthamoeba18S rRNA gene sequence type from an environmental isolate. Parasitology Research, 113, 2845-285010.1007/s00436-014-3945-2Search in Google Scholar PubMed

Marciano-Cabral F., Cabral G. 2003. Acanthamoeba spp. as agents of disease in humans. Clinical Microbiology Reviews, 16,273-30710.1128/CMR.16.2.273-307.2003Search in Google Scholar PubMed PubMed Central

Page F.C.1988. A New Key to Freshwater and Soil Gymnamoebae. Freshwater Biological Association, Cumbria, UK pp. 122Search in Google Scholar

Pérez de Galindo M.V., Galindo M., Dorta A., Guzmán de Rondón C., Wagner C., Vethencourt M.A., Nessi A., Bermúdez A., Pérez de Suárez E. 2012. Findings of Free-Living Amoebae of Acanthamoeba and Naegleria genus. A Venezuelan experience (Hallazgos de Amibas de Vida Libre de los géneros Acanthamoeba y Naegleria. Una experiencia venezolana). VITAE Academia Biomédica Digital, 51Search in Google Scholar

Pumidonming W., Koehsler M., Walochnik J. 2010. Acanthamoeba strains show reduced temperature tolerance after long-term axenic culture. Parasitology Research, 106, 553-55910.1007/s00436-009-1694-4Search in Google Scholar PubMed

Qvarnstrom Y., Nerad T.A., Visvesvara G.S. 2013. Characterization of a new pathogenic Acanthamoeba species, A. byersi n. sp., isolated from a human with fatal amoebic encephalitis. Journal of Eukaryotic Microbiology, 60, 626-63310.1111/jeu.12069Search in Google Scholar PubMed PubMed Central

Reyes-Batlle M., Todd C.D., Martín-Navarro C.M., López-Arencibia A., Cabello-Vilchez A.M., González A.C., Córdoba-Lanús E., Lindo J.F., Valladares B., Piñero J.E., Lorenzo-Morales J. 2014. Isolation and characterization of Acanthamoeba strains from soil samples in Gran Canaria, Canary Islands, Spain. Parasitology Research, 113, 1383-138810.1007/s00436-014-3778-zSearch in Google Scholar PubMed

Schuster F.L. 2002. Cultivation of pathogenic and opportunistic free-living amebas. Clinical Microbiology Reviews, 15, 342-35410.1128/CMR.15.3.342-354.2002Search in Google Scholar PubMed PubMed Central

Stothard D.R., Schroeder-Diedrich J.M., Awwad M.H., Gast R.J., Ledee D.R., Rodriguez-Zaragoza S., Dean C.L., Fuerst P.A., Byers T.J. 1998. The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. Journal of Eukaryotic Microbiology, 45,45-5410.1111/j.1550-7408.1998.tb05068.xSearch in Google Scholar PubMed PubMed Central

Tamura K., Peterson D., Peterson N., Stecher G., Nei M., Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731-273910.1093/molbev/msr121Search in Google Scholar PubMed PubMed Central

Todd C.D., Reyes-Batlle M., Martín-Navarro C.M., Dorta-Gorrín A., López-Arencibia A., Martínez-Carretero E., Piñero J.E., Valladares B., Lindo J.F., Lorenzo-Morales J. 2015. Isolation and genotyping of Acanthamoeba strains from soil sources from Jamaica, West Indies. Journal of Eukaryotic Microbiology, 62, 416-42110.1111/jeu.12197Search in Google Scholar PubMed

Visvesvara G.S., Moura H., Schuster F.L. 2007. Pathogenic and opportunistic free living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea. FEMS Immunology and Medical Microbiology Reviews, 50, 1-2610.1111/j.1574-695X.2007.00232.xSearch in Google Scholar PubMed

Wagner C., Dorta A., Pérez de Galindo M.V., Galindo M., Nessi A., Vethencourt A., Bandes A., Guzmán de Rondón C. 2011. Acanthamoeba spp. in a brain biopsie of a patient with metas-tastic adenocarcinoma (Hallazgo de Acanthamoeba spp en biopsia cerebral de un paciente con adenocarcinoma metastásico). Revista Instituto Nacional Higiene "Rafael Rangel", 42, 56-59Search in Google Scholar

Received: 2016-6-15

Revised: 2016-6-29

Accepted: 2016-6-30

Published Online: 2016-10-24

Published in Print: 2016-12-1

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Keywords for this article

Acanthamoeba spp; Venezuela; clinical samples; genotype