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Volume 61, Issue 2


Isolation and identification of Acanthamoeba spp. from thermal swimming pools and spas in Southern Brazil

Laura Fuhrich Fabres
  • Departamento de Microbiologia, Imunologia e Parasitologia, Instituto de Ciências Básicas da Saúde, Setor de Parasitologia, Universidade Federal do Rio Grande do Sul, Brasil
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/ Sayonara Peixoto Rosa dos Santos
  • Departamento de Microbiologia, Imunologia e Parasitologia, Instituto de Ciências Básicas da Saúde, Setor de Parasitologia, Universidade Federal do Rio Grande do Sul, Brasil
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/ Lisianne Brittes Benitez
  • Departamento de Biologia e Farmácia da Universidade de Santa Cruz do Sul, Santa Cruz, Rio Grande do Sul, Brasil
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/ Marilise Brittes Rott
  • Corresponding author
  • Departamento de Microbiologia, Imunologia e Parasitologia, Instituto de Ciências Básicas da Saúde, Setor de Parasitologia, Universidade Federal do Rio Grande do Sul, Sarmento Leite Street, N 500, Porto Alegre, Rio Grande do Sul, 90050-170, Brasil
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Published Online: 2016-03-30 | DOI: https://doi.org/10.1515/ap-2016-0031


Free-living amoebae (FLA) are widely distributed in soil and water. A few number of them are implicated in human disease: Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris and Sappinia diploidea. Species of Acanthamoeba can cause keratitis and brain infections. In this study, 72 water samples were taken from both hot tubs and thermal swimming pools in the city of Porto Alegre, RS, Brazil, to determine the presence of Acanthamoeba in the water as well as perform the phenotypic and genotypic characterization of the isolates. The identification of the isolates was based on the cysts morphology and PCR amplification using genus-specific oligonucleotides. When the isolates were submitted to PCR reaction only 8 were confirmed as belonging to the genus Acanthamoeba. The sequences analysis when compared to the sequences in the GenBank, showed genotype distribution in group T3 (12,5%), T5 (12,5%), T4 (25%) and T15 (50%). The results of this study confirmed the presence of potentially pathogenic isolates of free living amoebae in hot swimming pool and spas which can present risks to human health.

Key words: Acanthamoeba; thermal swimming-pool; hot tubs; genotypes; environmental isolates


  • Alves J.M.P., Gusmão C.X., Teixeira M.M.G., Freitas D., Foronda A.S., Affonso H.T. 2000. Random amplified polymorphic DNA profiles as a tool for the characterization of Brazilian keratitis isolates of the genus Acanthamoeba. Brazilian Journal of Medical and Biological Research, 33, 19–26. DOI: http://dx.doi.org/10.1590/S0100-879X2000000100003Google Scholar

  • Alves D.D., Moraes A.S., Nitz N., Oliveira M.G., Hecht M.M., Gurgel-Gonçalves R., Cuba C.A. 2012. Occurrence and characterization of Acanthamoeba similar to genotypes T4, T5, and T2/T6 isolated from environmental sources in Brasília, Federal District, Brazil. Experimental Parasitoly, 131, 239–244. DOI: 10.1016/j.exppara.2012.04.011CrossrefGoogle Scholar

  • APHA 2005. Standard Methods for the Examination of Water and Wastewater. Washington, DC: APHA, WEF and AWWAGoogle Scholar

  • Booton G.C., Rogerson A., Bonilla T.D., Seal D.V., Kelly D.J., Beat-tie T.K., Tomlinson A., Lares-Villa F., Fuerst P.A., Byers T.J. 2004. Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis. Journal of Eukaryotic Microbiology, 51, 192–200. DOI: 10.1111/j.1550-7408.2004.tb00545.xCrossrefGoogle 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, 1689–1693. DOI: 10.1128/JCM.43.4.1689-1693.2005CrossrefGoogle Scholar

  • Booton, G.C., Joslin C.E., Shoff M., Ey T.U., Kelly D.J., Fuerst P.A. 2009. Genotypic identification of Acanthamoeba sp. isolates associated with an outbreak of Acanthamoeba keratitis. Cornea 28, 673–676. DOI: 10.1097/ICO.0b013e31819342a7CrossrefGoogle Scholar

  • Carlesso A.M., Artuso G.L., Caumo K., Rott M.B. 2010. Potentially pathogenic Acanthamoeba isolated from a hospital in Brazil. Current Microbiology, 60, 185–190. DOI: 10.1007/s00284009-9523-7CrossrefGoogle Scholar

  • Caumo K., Frasson A.P., Pens C.J., Panatieri L.F., Frazzon A.P.G., Rott M.B. 2009. Potentially pathogenic Acanthamoeba in swimming pools: a survey in the southern Brazilian city of Porto Alegre. Annals of Tropical Medicine Parasitology, 103, 477–485. DOI: 10.1179/136485909X451825CrossrefGoogle Scholar

  • Caumo K., Rott M.B. 2011. Acanthamoeba T3, T4 and T5 in swim-ming-pool waters from Southern Brazil. Acta Tropica, 117, 233–235. DOI: 10.1016/j.actatropica.2010.12.008CrossrefGoogle Scholar

  • Corsaro D., Venditti D. 2010. Phylogenetic evidence for a new genotype of Acanthamoeba (Amoebozoa Acanthamoebida). Parasitology Research, 7, 233–8. DOI: 10.1007/s00436-0101870-6CrossrefGoogle 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, 2481–2490. DOI: 10.1007/s00436-015-4445-8CrossrefGoogle Scholar

  • Di Cave D.D., Monno R., Bottalico P., Guerriero S., D’Amelio S., D’Orazi C., Berrilli F. 2009. Acanthamoeba T4 and T15 genotypes associated with keratitis infections in Italy. European Journal of Clinical Microbioly and Infectious Diseases, 28, 607–612. DOI: 10.1007/s10096-008-0682-4CrossrefGoogle Scholar

  • Di Cave D.D., D’alfonso R., Comlavi D., Orazi C.D., Monno R., Berrili F. 2014. Genotypic heterogeneity based on 18S-rRNA gene sequences among Acanthamoeba isolates from clinical samples in Italy. Experimental Parasitology, 145, 46–49. DOI: 10.1016/j.exppara.2014.05.009CrossrefGoogle Scholar

  • Duarte J.L., Furst C., Klisiowicz D.R., Klassen G., Costa A.O. 2013. Morphological, genotypic, and physiological characterization of Acanthamoeba isolates from keratitis patients and the domestic environment in Vitoria, Espirito Santo, Brazil. Experimental Parasitology, 135, 9–14. DOI: 10.1016/j.exppara. 2013.05.013CrossrefGoogle Scholar

  • Edagawa A., Kimura A., Kawabuchi-Kurata T., Kusuhara Y., Karanis P. 2009. Isolation and genotyping of potentially pathogenic Acanthamoeba and Naegleria species from tap-water sources in Osaka, Japan. Parasitology Research, 105, 1109– 1117. DOI: 10.1007/s00436-009-1528-4CrossrefGoogle Scholar

  • Flint J.A., Dobson P.J., Robinson B.S. 2003. Genetic analysis of forty isolates of Acanthamoeba group III by multilocus isoenzyme electrophoresis. Acta Protozoologica, 42, 317–324Google Scholar

  • Gast R.J. 2001. Development of an Acanthamoeba-specific reverses dot-blot and the discovery of a new ribotype. Journal of Eukaryotic Microbiology, 48, 609–15. DOI: 10.1111/j.15507408.2001.tb00199.xCrossrefGoogle Scholar

  • Gianinazzi C., Schild M., Zumkehr B., Wuthrich F., Nuesch I., Ryter R., Schurch N., Gottstein B., Muller N. 2010. Screening of Swiss hot spring resorts for potentially pathogenic free-living amoebae. Experimental Parasitology, 126, 45–53. DOI: 10.1016/j.exppara.2009.12.008CrossrefGoogle Scholar

  • Greub G., Raoult D. 2004. Microorganisms resistant to free-living amoebae. Clinical Microbiology Reviews, Washington 17, 413–433. DOI: 10.1128/CMR.17.2.413-433CrossrefGoogle Scholar

  • Hewett 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. Acta Protozoologica, 42, 325–9Google 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, 357–67. DOI: 10. 1046/j.1462-2920.1999.00045.xCrossrefGoogle Scholar

  • Huang S.W., Hsu B.M. 2010. Isolation and identification of Acanthamoeba from Taiwan spring recreation areas using culture enrichment combined with PCR. Acta Tropica, 115, 282–287. DOI: 10.1016/j.actatropica.2010.04.012CrossrefGoogle Scholar

  • Kao P.M., Hsu B.M., Chen N.H., Huang K.H., Huang S.W., King K.L., Chiu Y.C. 2012a. Isolation and identification of Acanthamoeba species from thermal spring environmental in southern Taiwan. Experimental Parasitology, 130, 354–358. DOI: 10.1016/j.exppara.2012.02.008CrossrefGoogle Scholar

  • Kao P.M., Hsu B.M., Chen N.H., Huang K.H., Huang C.C., Ji D.D., Chen J.S., Lin W.C., Huang S.W., Chiu Y.C. 2012b. Molecular detection and comparison of Acanthamoeba genotypes in different functions of watersheds in Taiwan. Environmental Monitoring and Assessment, 184, 4335–4344. DOI: 10.1007/s10661-011-2267-4CrossrefGoogle Scholar

  • Khan N.A., Jarroll E.L., Paget T.A. 2001. Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays. Current Microbiology, 43, 204–208. DOI: 10.1007/s002840010288CrossrefGoogle Scholar

  • Khan N.A., Tareen N.K. 2003. Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorization of Acanthamoeba strains. Folia Parasitology, 50, 97–104. DOI: 10.14411/fp.2003.017CrossrefGoogle Scholar

  • Khan N.A. 2006. Acanthamoeba: Biology and increasing importance in human health. FEMS Microbiological Reviews, 30, 564–595. DOI: 10.1111/j.1574-6976.2006.00023.xCrossrefGoogle Scholar

  • Kiss C., Barna Z., Vargha M., Torok J.K. 2014. Incidence and molecular diversity of Acanthamoeba species isolated from public baths in Hungary. Parasitology Research, 113, 2551–2557. DOI: 10.1007/s00436-014-3905-xCrossrefGoogle Scholar

  • Koehsler M., Leitsch D., Ducheˆne M., Nagl M., Walochnik J. 2009. Acanthamoeba castellanii, growth on human cell layers reactivates attenuated properties after prolonged axenic culture. FEMS Microbiology Letters, 299, 121–127. DOI, 10.1111/ j.1574-6968.2009.01680.xCrossrefGoogle Scholar

  • Ledee D.R., Hay J., Byers T.J., Seal D.V., Kirkness C.M. 1996. Acanthamoeba griffini. Molecular characterization of a new corneal pathogen. Investigative Ophthalmology & Visual Science, 37, 544–550Google Scholar

  • Ledee D.R., Iovieno A., Miller D., Mandal N., Diaz M., Fell J., Fini M.E., Alfonso E.C. 2009. Molecular identification of T4 and T5 genotypes in isolates from Acanthamoeba keratitis patients. Journal of Clinical Microbiology, 47, 1458–1462. DOI: 10.1128/JCM.02365-08CrossrefGoogle Scholar

  • Liang S.Y., Ji D.R., Hsia K.T., Hung C.C., Sheng W.H., Hsu B.M., Chen J.S., Wu M.H., Lai C.H., Ji D.D. 2010. Isolation and identification of Acanthamoeba species related to amoebic encephalitis and nonpathogenic free-living amoeba species from the rice field. Journal of Applied Microbiology, 109, 1422– 1429. DOI: 10.1111/j.1365-2672.2010.04779.xCrossrefGoogle Scholar

  • Maghsood A.H., Sissons J., Rezaian M., Nolder D., Warhurst D., Khan N.A. 2005. Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates. Journal of Medical Microbiology, 54, 755–759. DOI: 10.1099/jmm.0.45970-0CrossrefGoogle Scholar

  • Magliano A.C.M., Silva F.M., Teixeira M.M.G., Alfieri S.C. 2009. Genotyping, physiological features and proteolytic activities of a potentially pathogenic Acanthamoeba sp. isolated from tap water in Brazil. Experimental Parasitology, 12, 231–235. DOI: 10.1016/j.exppara.2009.07.006CrossrefGoogle Scholar

  • Magliano A.C., Teixeira M.M., Alfier, S.C. 2012. Revisiting the Acanthamoeba species that form star-shaped cysts (genotypes T7, T8, T9, and T17): characterization of seven new Brazilian environmental isolates and phylogenetic inferences. Parasitology, 139, 45–52. DOI: 10.1017/S0031182011001648CrossrefGoogle Scholar

  • Magnet A., Henriques-Gil N., Galván-Diaz A.L., Izquiedo F., Fenoy S., Del Aguila C. 2014. Novel Acanthamoeba 18S rRNA gene sequence type from an envirionmental isolate. Parasitology Research, 113, 2845–2850. DOI: 10.1007/s00436014-3945-2CrossrefGoogle Scholar

  • Marciano-Cabral F., Cabral G. 2003 Acanthamoeba spp. as agents of disease in humans. Clinical Microbiology Reviews, 16, 273–307. DOI: 10.1128/CMR.16.2.273-307.2003CrossrefGoogle Scholar

  • Mathers W.D., Nelson S.E., Scott N.E., Lane J.L., Wilson M.E., Allen R.C. Folberg R. 2000. Confirmation of confocal microscopy diagnosis of Acanthamoeba keratitis using polymerase chain reaction analysis. Archives of Ophthalmology, 118, 178–183. DOI:10.1001/archopht.118.2.178CrossrefGoogle Scholar

  • Nagyová V., Nagy A., Timko J. 2010. Morphological, physiological and molecular biological characterisation of isolates from first cases of Acanthamoeba keratitis in Slovakia. Parasitology Research, 106, 861–872. DOI: 10.1007/s00436-010-1731-3CrossrefGoogle Scholar

  • Nuprasert W., Putaporntip C., Pariyakanok L., Jongwutiwes S. 2010. Identification of a novel T17 genotype of Acanthamoeba from environmental isolates and T10 genotype causing keratitis in Thailand. Journal of Clinical Microbiology, 48, 4636–40. DOI: 10.1128/JCM.01090-10CrossrefGoogle Scholar

  • Page F.C. 1967. Re-definition of the genus Acanthamoeba with descriptions of three species. The Journal of Protozoology, 14, 709–724. DOI: 10.1111/j.1550-7408.1967.tb02066.xCrossrefGoogle Scholar

  • Page FC 1988. A New Key to Freshwater and Soil Amoebae. Freshwater Biological Association Scientific Publications, Cumbria. DOI: 10.1002/iroh.19780630231CrossrefGoogle Scholar

  • Pussard M., Pons R. 1977. Morphologie de la paroi kystique et taxonomie du genre Acanthamoeba (Protozoa, Amoebida). Protistologica, 13, 557–598Google Scholar

  • 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, 626–633. DOI: 10.1111/ jeu.12069CrossrefGoogle Scholar

  • Salah M., Iciar M. 1997. Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. Nucleic Acids Research, Oxford, 25, 4692–4693. DOI: 10.1093/nar/ 25.22.4692CrossrefGoogle Scholar

  • Schroeder J.M., Booton G.C., Hay J., Niszl I.A., Seal D.V., Markus M.B., Fuerst P.A., Byers T.J. 2001. Use of subgenic 18S ribosomal DNA PCR and sequencing for genus and genotype identification of Acanthamoebae from humans with keratitis and from sewage sludge. Journal of Clinical Microbiology, 39, 1903–1911. DOI: 10.1128/JCM.39.5.1903-1911.2001CrossrefGoogle Scholar

  • Schuster F.L., Visvesvara G.S. 2004. Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals. International Journal of Parasitology, 34, 1001–1027. DOI: 10.1016/j.ijpara.2004.06.004CrossrefGoogle Scholar

  • 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 Eukaryotic Microbiology, 45, 45–54Google Scholar

  • Tanveer T., Hameed A., Muazzam A.G., Jung S.Y., Gul A., Matin A. 2013. Isolation and molecular characterization of potentially pathogenic Acanthamoeba genotypes from diverse resources including household driking water from Khyber Paktunkhwa, Pakistan. Parasitology Research, 112, 2925–2932. DOI: 10.1007/s00436-013-3465-5CrossrefGoogle Scholar

  • Trabelsi H., Dendana F., Sellami A., Sellami H., Cheikhrouhou F., Neji S., Makni F., Ayadi A. 2012. Pathogenic free-living amoebae: Epidemiology and clinical review. Pathologie Biologie, 60, 399–405. DOI: 10.1016/j.patbio.2012.03.002CrossrefGoogle Scholar

  • Tsvetkova N., Schild M., Panaiotv S., Kurdova-Mintcheva R., Gottstein B., Walochnik J. 2004. The identification of free-living environmental isolates of amoebae from Bulgaria. Parasitology Research, 92, 405–413. DOI: 10.1007/s00436-0031052-xCrossrefGoogle Scholar

  • 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, 50, 1–26. DOI: http://dx.doi.org/10.1111/j.1574-695X.2007.00232.xGoogle Scholar

  • Visvesvara G., Schuster F. 2008a. Opportunistic free-living amebae, Part I. Clinical Microbiology Newsletter, 30, 151–158. DOI: 10.1016/j.clinmicnews.2008.09.004CrossrefGoogle Scholar

  • Visvesvara G., Schuster F. 2008b Opportunistic free-living amebae, Part II. Clinical Microbiology Newsletter, 30, 159–166. DOI: 10.1016/j.clinmicnews.2008.10.001CrossrefGoogle Scholar

  • Walochnik J., Obwaller A., Aspöck H. 2000. Correlations between morphological, molecular, biological and physiological characteristics in clinical and nonclinical isolates of Acanthamoeba spp. Applied Environmental Microbiology, 66, 4408–4413. DOI: 10.1128/AEM.66.10.4408-4413.2000CrossrefGoogle Scholar

  • Winck M.A., Caumo K., Rott M.B. 2011. Prevalence of Acanthamoeba from tap water in Rio Grande do Sul, Brazil. Current Microbiology, 63, 464–469. DOI: 10.1007/s00284-011-0003-5CrossrefGoogle Scholar

  • Zhang Y., Sun X., Wang Z., Li R., Luo S., Jin X., Deng S., Chen W. 2004. Identification of 18S ribosomal DNA genotype of Acanthamoeba from patients with keratitis in North China. Investigative Ophthalmology & Visual Science, 45, 1904–1907. DOI: 10.1167/iovs.03-1073CrossrefGoogle Scholar

About the article

Received: 2015-04-04

Revised: 2015-09-29

Accepted: 2015-11-25

Published Online: 2016-03-30

Published in Print: 2016-06-01

Citation Information: Acta Parasitologica, Volume 61, Issue 2, Pages 221–227, ISSN (Online) 1896-1851, ISSN (Print) 1230-2821, DOI: https://doi.org/10.1515/ap-2016-0031.

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