Advertisement

Acta Parasitologica

, Volume 61, Issue 4, pp 796–801 | Cite as

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-MoralesEmail author
Article

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 Acanthamoeba 18S 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 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 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, 63–68Google Scholar
  2. 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, 157–163Google Scholar
  3. 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, 1689–1693CrossRefGoogle Scholar
  4. 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, 2509–2513CrossRefGoogle Scholar
  5. 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 2010Google Scholar
  6. 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, 2481–2490CrossRefGoogle Scholar
  7. 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, 185–197Google Scholar
  8. 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, 135–142PubMedGoogle Scholar
  9. 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, 42–54Google Scholar
  10. 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, 84–91Google Scholar
  11. Gast R.J. 2001. Development of an Acanthamoeba-specific reverse dot-blot and the discovery of a new ribotype. Journal of Eukaryotic Microbiology, 48, 609–615CrossRefGoogle Scholar
  12. 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, 498–504CrossRefGoogle Scholar
  13. Geisen S., Fiore-Donno A.M., Walochnik J., Bonkowski M. 2014. Acanthamoeba everywhere: high diversity of Acanthamoeba in soils. Parasitology Research, 113, 3151–3158CrossRefGoogle Scholar
  14. Hewet M.K., Robinson B.S., Monis P.T., Saint C.P. 2003. Identification of a new Acanthamoeba 18S rRNA gene sequence type, corresponding to the species Acanthamoeba jacobsi Sawyer, Nerad and Visvesvara, 1992 (Lobosea: Acanthamoebidae). Acta Protozoologica, 42, 325–329Google Scholar
  15. 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, 357–367CrossRefGoogle Scholar
  16. Khan N.A. 2006. Acanthamoeba: biology and increasing importance in human health. FEMS Microbiology Reviews, 30, 564–595CrossRefGoogle Scholar
  17. Khan N.A. 2009. Acanthamoeba: Biology and Pathogenesis. Caister Academic Press: Norfolk, Great Britain, pp. 290Google Scholar
  18. 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, 1119–1206CrossRefGoogle Scholar
  19. 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, 1069–1072CrossRefGoogle Scholar
  20. Lorenzo-Morales J., Khan N.A., Walochnik J. 2015. An update on Acanthamoeba keratitis: diagnosis, pathogenesis and treatment. Parasite, 22, 10CrossRefGoogle Scholar
  21. 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, 27–33Google Scholar
  22. 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, 490–513CrossRefGoogle Scholar
  23. Magnet A., Henriques-Gil N., Galván-Diaz A.L., Izquierdo F., Fenoy S., del Aguila C. 2014. Novel Acanthamoeba 18S rRNA gene sequence type from an environmental isolate. Parasitology Research, 113, 2845–2850CrossRefGoogle Scholar
  24. Marciano-Cabral F., Cabral G. 2003. Acanthamoeba spp. as agents of disease in humans. Clinical Microbiology Reviews, 16, 273–307CrossRefGoogle Scholar
  25. Page F.C. 1988. A New Key to Freshwater and Soil Gymnamoebae. Freshwater Biological Association, Cumbria, UK pp. 122Google Scholar
  26. 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, 51Google Scholar
  27. Pumidonming W., Koehsler M., Walochnik J. 2010. Acanthamoeba strains show reduced temperature tolerance after long-term axenic culture. Parasitology Research, 106, 553–559CrossRefGoogle Scholar
  28. 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, 626–633CrossRefGoogle Scholar
  29. 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, 1383–1388CrossRefGoogle Scholar
  30. Schuster F.L. 2002. Cultivation of pathogenic and opportunistic freeliving amebas. Clinical Microbiology Reviews, 15, 342–354CrossRefGoogle Scholar
  31. 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 TJ. 1998. The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. Journal of Eukaryotic Microbiology, 45, 45–54CrossRefGoogle Scholar
  32. 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, 2731–2739CrossRefGoogle Scholar
  33. 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, 416–421CrossRefGoogle Scholar
  34. 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, 1–26CrossRefGoogle Scholar
  35. 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 metastastic adenocarcinoma (Hallazgo de Acanthamoeba spp en biopsia cerebral de un paciente con adenocarcinoma metastásico). Revista Instituto Nacional Higiene “Rafael Rangel”, 42, 56–59Google Scholar

Copyright information

© Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2016

Authors and Affiliations

  • Carolina Wagner
    • 1
    • 2
  • María Reyes-Batlle
    • 1
  • María Alejandra Vethencourt Ysea
    • 2
  • Mónica V. Galindo Pérez
    • 2
  • Carmen Guzmán de Rondón
    • 2
  • Anaibeth J. Nessi Paduani
    • 2
  • Angelyseb Dorta Pérez
    • 2
  • Atteneri López-Arencibia
    • 1
  • Ines Sifaoui
    • 1
    • 3
  • María Virginia Pérez de Galindo
    • 2
  • Eva Pérez de Suárez
    • 2
  • Enrique Martínez-Carretero
    • 1
  • Basilio Valladares
    • 1
  • José E. Piñero
    • 1
  • Jacob Lorenzo-Morales
    • 1
    Email author
  1. 1.University Institute of Tropical Diseases and Public Health of the Canary IslandsUniversity of La LagunaTenerife, Canary IslandsSpain
  2. 2.Laboratorio de Amibiasis-Cátedra de Parasitología, Escuela de Bioanálisis, Facultad de MedicinaUniversidad Central de VenezuelaCaracasVenezuela
  3. 3.Laboratoire Matériaux-Molécules et Applications, IPESTUniversity of CarthageLa MarsaTunisia

Personalised recommendations