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Isolation and molecular characterization of Acanthamoeba and Balamuthia mandrillaris from combination shower units in Costa Rica

Abstract

Free living amoebae (FLA) are ubiquitous protozoa, which may behave as parasites under certain conditions. Four genera are recognized as causal agents of infections in humans and animals: Naegleria, Sappinia, Acanthamoeba and Balamuthia. This work determines the presence of FLA in combination shower units and employs molecular biology for the characterization of isolates. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of Acanthamoeba genotype T4 in 30 % of the units sampled. In addition to Acanthamoeba cysts, trophozoites with morphological characteristics similar to Balamuthia were identified. PCR assay using the mitochondrial 16S rRNA gene as a target confirmed the identification of the amoeba as Balamuthia mandrillaris. Up to date, this is the first report of the isolation of B. mandrillaris in Central America and the fifth report worldwide.

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References

  1. Adl SM, Simpson AGB, Farmer MA, Andersen RA, Barta JR, Bowser SS, Brugerolle G, Fensome RA, Fredericq S, James TY, Karpov S, Kugrens P, Krug J, Lane CE, Lewis LA, Lodge J, Lynn DH, Mann DG, McCourt RM, Mendoza L, Moestrup O, Mozley-Standridge SE, Nerad TA, Shearer CA, Smirnov AV, Spiegel FW, Taylor MF (2005) The new higher level classification of eukaryotes with emphasis on the taxonomy of protists. J Eukaryot Microbiol 52(5):399–451

  2. Amaral Zettler LA, Nerad TA, O’Kelly CJ, Peglar MT, Gillevet PM, Silberman JD, Sogin ML (2000) A molecular reassessment of the leptomyxid amoebae. Protist 151:275–282

  3. Bier J, Sawyer T (1990) Amoeba isolated from laboratory eyewash stations. Curr Microbiol 20(5):349–350

  4. Booton GC, Kelly DJ, Chu YW, Seal DV, Houang E, Lam DSC, Byers TJ, Fuerst PA (2002) 18S ribosomal DNA typing and tracking of Acanthamoeba species isolates from corneal scrape specimens, contact lenses, lens cases, and home water supplies of Acanthamoeba keratitis patients in Hong Kong. J Clin Microbiol 40:1621–1625

  5. Cabello-Vílchez AM, Reyes-Batlle M, Montalbán-Sandoval E, Martín-Navarro CM, López-Arencibia A, Elias-Letts R, Guerra H, Gotuzzo E, Martínez-Carretero E, Piñero JE, Maciver SK, Valladares B, Lorenzo-Morales J (2014) The isolation of Balamuthia mandrillaris from environmental sources from Peru. Parasitol Res 2014 Apr 30. [Epub ahead of print]

  6. Chinchilla M, Castro E, Alfaro M, Portilla E (1979) Amebas de vida libre productoras de meningoencefalitis. Primeros hallazgos en Costa Rica. Rev Latinoam Microbiol 21:135–142

  7. Corsaro D, Venditti D (2010) Phylogenetic evidence for a new genotype of Acanthamoeba (Amoebozoa Acanthamoebida). Parasitol Res 7:233–238

  8. De Jonckheere JF (2007) Molecular identification of free-living amoebae of the Vahlkampfiidae and Acanthamoebidae isolated in Arizona (USA). Eur J Protistol 43:9–15

  9. Dunnebacke TH, Schuster FL, Yagi S, Booton GC (2004) Balamuthia mandrillaris from soil samples. Microbiol 150:2837–2842

  10. Echandi L, González D, Marín R (1994) Aislamiento de amebas de vida libre capaces de producir meningoencefalitis. Rev Méd de Costa Rica y Centroamérica LXI 526:11–14

  11. Gast RJ (2001) Development of an Acanthamoeba-specific reverses dot-blot and the discovery of a new ribotype. J Eukaryot Microbiol 48:609–615

  12. Gelman BB, Rauf SJ, Nader R, Popov V, Bokowski J, Chaljub G, Nauta HW (2001) Amoebic encephalitis due to Sappinia diploidea. JAMA 285:2450–2451

  13. Hewett MK, Robinson BS, Monis PT, Saint CP (2003) Identification of a new Acanthamoeba 18S rRNA gene sequence type, corresponding to the species Acanthamoeba jacobsi Sawyer. Acta Protozool 42:325–329

  14. Horn M, Fritsche TR, Gautom RK, Schleifer KH, Wagner M (1999) Novel bacterial endosymbionts of Acanthamoeba spp. related to the Paramecium caudatum symbiont Caedibacter caryophilus. Environ Microbiol 1:357–367

  15. Khan NA (2006) Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev 30:564–595

  16. Kiss C, Barna Z, Vargha M, Török JK (2014) Incidence and molecular diversity of Acanthamoeba species isolated from public baths in Hungary. Parasitol Res. 2014 Apr 30. [Epub ahead of print]

  17. Lass A, Szostakowska B, Idzińska A, Chomicz L (2014) The first genotype determination of Acanthamoeba potential threat to human health, isolated from natural water reservoirs in Poland. Parasitol Res. 2014 Apr 27. [Epub ahead of print].

  18. Lorenzo-Morales J, Cabello-Vílchez AM, Martín-Navarro CM, Martínez-Carretero E, Piñero JE, Valladares B (2013) Is Balamuthia mandrillaris a public health concern worldwide? Trends Parasitol 29:483–488

  19. Matin A, Siddiqui R, Jayasekera S, Khan NA (2008) Increasing importance of Balamuthia mandrillaris. Clin Microbiol Rev 21:435–448

  20. Michel R, Just HM (1984) Acanthamoebae, Naegleria and other free-living Amoebae in cooling and rinsing water of dental treatment units. Zbl Bakteriol Mikrobiol Hyg 179:56–72

  21. Morales JA, Chaves AJ, Visvesvara GS, Dubey JP (2006) Naegleria fowleri associated encephalitis in a cow from Costa Rica. Vet Parasitol 139(1–3):221–223

  22. Nacapunchai D, Lamom C, Ruangsittichai C, Sriwichai P (1999) Isolation of free-living amebae from soil and water resources in Thailand. J Trop Med Parasitol 22:22–26

  23. Niyyati M, Lorenzo-Morales J, Rezaeian M, Martin-Navarro CM, Haghi AM, Maciver SK, Valladares B (2009) Isolation of Balamuthia madrillaris from urban dust, free of known infectious involvement. Parasitol Res 106:279–281

  24. 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. J Clin Microbiol 48:4636–4640

  25. Page FC (1988) A new key to freshwater and soil gymnamoebidae. Freshwater Biological Association, Ambleside, 121 pp

  26. Paszko-Kolva C, Yamamoto H, Shahamat M, Sawyer TK, Morris G, Colwel RR (1991) Isolation of amoebae and Pseudomonas and Legionella spp. from eyewash stations. Appl Environ Microbiol 57(1):163–167

  27. Qvarnstrom Y, Nerad TA, Visvesvara GS (2013) Characterization of a new pathogenic Acanthamoeba species, A. byers in. sp., isolated from a human with fatal amoebic encephalitis. J Eukaryot Microbiol 60:626–633

  28. Reyes-Batlle M, Todd CD, Martín-Navarro CM, López-Arencibia A, Cabello-Vílchez AM, González AC, Córdoba-Lanús E, Lindo JF, Valladares B, Piñero JE, Lorenzo- Morales J (2014) Isolation and characterization of Acanthamoeba strains from soil samples in Gran Canaria, Canary Islands, Spain. Parasitol Res 113(4):1383–1388

  29. Schuster FL, Visvesvara GS (2004) Free-living amoebae as opportunistic and non- opportunistic pathogens of humans and animals. Int J Parasitol 34:1001–1027

  30. Schuster FL, Glaser C, Gilliam S, Visvesvara GS (2001) Survey of sera from encephalitis patients for Balamuthia mandrillaris antibody. J Eukaryot Microbiol. Suppl 10S- 12S

  31. Schuster FL, Dunnebacke TH, Booton GC, Yagi S, Kohlmeier CK, Glaser C, Vugia D, Bakardjiev A, Azimi P, Maddux-Gonzalez M, Martinez AJ, Visvesvara GS (2003) Environmental isolation of Balamuthia mandrillaris associated with a case of amebic encephalitis. J Clin Microbiol 41:3175–3180

  32. Schuster FL, Yagi S, Gavali S, Michelson D, Raghavan R, Blomquist I, Glastonbury C, Bollen AW, Scharnhorst D, Reed SL, Kuriyama S, Visvesvara GS, Glaser CA (2009) Under the radar: Balamuthia amebic encephalitis. Clin Infect Dis 48:879–887

  33. Stothard DR, Schroeder-Diedrich JM, Awwad MH, Gast RJ, Ledee DR, Rodriguez- Zaragoza S, Dean CL, Fuerst PA, Byers TJ (1998) The evolutionary history of the genus Acanthamoebaand the identification of eight new 18S rRNA gene sequence types. J Eukaryot Microbiol 45:45–54

  34. 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. Mol Biol Evol 28:2731–2739

  35. Tanveer T, Hameed A, Muazzam AG, Jung SY, Gul A, Matin A (2013) Isolation and molecular characterization of potentially pathogenic Acanthamoeba genotypes from diverse water resources including household drinking water from Khyber Pakhtunkhwa. Pak Parasitol Res 112:2925–2932

  36. Trabelsi H, Dendana F, Sellami A, Cheikhrouhou F, Neji S, Makni F, Ayadi A (2012) Pathogenic free-living amoebae: epidemiology and clinical review. Pathol Biol (Paris) 60:399–405

  37. Tyndall RL, Lyle MM, Ironside KS (1987) The presence of free-living amoebae in portable and stationary eyewash stations. Am Ind Hyg Assoc J 48:933–934

  38. Visvesvara GS, Moura H, Schuster FL (2007) Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri and Sappinia diploidea. FEMS Immunol Med Microbiol 50:1–26

  39. Wagner CM, Dorta PA, MV P d G, Galindo PMV, Nessi PA, Venthecourt MA, Bandes A, CT G d R (2011) Hallazgo de Acanthamoeba spp en biopsia cerebral de un paciente con adenocarcinoma metastásico. RINHRR 42(2):56–59

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Acknowledgments

This study was supported by project 803-B4-050, Vicerrectoría de Investigación, University of Costa Rica. This work was supported by the grants RICET (project no. RD12/0018/0012of the programme of Redes Temáticas de Investigación Cooperativa, FIS), Spanish Ministry of Health, Madrid, Spain and the Project FIS PI10/01298 “Protozoosis emergentes por amebas de vida libre: aislamiento y caracterización molecular, identificación de cepas transportadoras de otros agentes patógenos y búsqueda de quimioterapias efectivas”, 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. MRB was funded by CEI Canarias, Campus Atlántico Internacional and Becas Fundación Cajacanarias para Postgraduados 2014. JLM was supported by the Ramón y Cajal Subprogramme from the Spanish Ministry of Economy and Competivity RYC-2011-08863.

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Correspondence to Elizabeth Abrahams-Sandí.

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Retana-Moreira, L., Abrahams-Sandí, E., Cabello-Vílchez, A.M. et al. Isolation and molecular characterization of Acanthamoeba and Balamuthia mandrillaris from combination shower units in Costa Rica. Parasitol Res 113, 4117–4122 (2014). https://doi.org/10.1007/s00436-014-4083-6

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Keywords

  • Free-living amoebae
  • Acanthamoeba
  • Balamuthia mandrillaris
  • Costa Rica