Parasitology Research

, Volume 95, Issue 4, pp 273–277 | Cite as

Isolation and identification of pathogenic Acanthamoeba strains in Tenerife, Canary Islands, Spain from water sources

  • Jacob Lorenzo-Morales
  • Antonio Ortega-RivasEmail author
  • Pilar Foronda
  • Enrique Martínez
  • Basilio Valladares
Original Paper


A comprehensive survey to document the presence of free-living amoebae of the genus Acanthamoeba was conducted in tap water and sea water sources related to human environments in Tenerife, Canary Islands, Spain. Acanthamoeba identification was based on the morphology of cyst and trophozoite forms and PCR amplification with a genus-specific primer pair. The pathogenic potential of Acanthamoeba isolates was characterized by temperature and osmotolerance assays and PCR reactions with two primer pairs related to Acanthamoeba pathogenesis. The results demonstrate the presence of potentially pathogenic strains in both sources. Thus, some of the amoebae in these aquatic habitats can act as opportunistic pathogens, could play a role in the diseases of aquatic organisms, and may present a risk to human health.


Keratitis Canary Island Acanthamoeba Isolate Acanthamoeba Strain Species Specific Prime Pair 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported by Dirección General de Universidades del Gobierno de Canarias.


  1. Alsam S, Kim KS, Stins M, Rivas AO, Sissons J, Khan NA (2003) Acanthamoeba interactions with human brain microvascular endothelial cells. Microb Pathog 35:235–241Google Scholar
  2. Alves JM., Gusmao CX, Teixeira MM, Freitas D, Foronda A, Affonso HT (2000) Random amplified polymorphic DNA profiles as a tool for the characterization of Brazilian keratitis isolates of the genus Acanthamoeba. Braz J Med Biol Res 33:19–26Google Scholar
  3. Armstrong M (2000) The pathogenesis of human Acanthamoeba infection. Infect Dis Rev 2:65–73Google Scholar
  4. Booton GC, Kelly DJ, Chu YW, Seal DV, Houang E, Lam DS, 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 40:1621–1625Google Scholar
  5. Chappell CL, Wright JA, Coletta M, Newsome AL (2001) Standarized method of measuring Acanthamoeba in sera from healthy human subjects. Clin Diag Lab Immunol 8:724–730Google Scholar
  6. Ettinger MR, Webb SR, Harris SA, McIninch SP, Garman G, Brown BL (2003) Distribution of free-living amoebae in James River, Virginia, USA. Parasitol Res 89:6–15Google Scholar
  7. Hadas E, Mazur T (1993) Biochemical markers of pathogenicity and virulence of Acanthamoeba sp. strains. Parasitol Res 79:696–698Google Scholar
  8. Hong YC, Kong HH, Ock MS, Kim IN, Chung DI (2000) Isolation and characterization of a cDNA encoding a subtilisin-like serin proteinase (AhSUB) from Acanthamoeba healyi. Mol Biol Parasitol 111:441–446Google Scholar
  9. Howe DK, Vodkin MH, Novak RJ, Visvesvara G, McLaughlin GL (1997) Identification of two genetic markers that distinguish pathogenic and nonpathogenic strains of Acanthamoeba spp. Parasitol Res 83:345–348Google Scholar
  10. Khan NA (2003) Pathogenesis of Acanthamoeba infections. Microb Pathog 34:277–285Google Scholar
  11. Khan NA, Tareen NK (2003) Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorisation of Acanthamoeba strains. Folia Parasitol 50:97–104Google Scholar
  12. Khan NA, Jarroll EL, Paget TA (2001) Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays. Curr Microbiol 43:204–208Google Scholar
  13. Kong HH, Shin JY, Yu HS, Kim J, Hahn TW, Hahn YH, Chung DI (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. J Clin Microbiol 40:1199–1206Google Scholar
  14. Levine S, Goldstein AE, Dahdouh M, Blank P, Hoffman C, Gropper CA (2001) Cutaneous Acanthamoeba in a patient with AIDS: a case study with a review of new therapy. Cutis 67:377–380Google Scholar
  15. Ma P, Visvesvara G, Martinez A, Theodore FH, Dagget P, Sawyer T (1990) Naegleria and Acanthamoeba infections: review. Rev Infect Dis 12:490–513Google Scholar
  16. Marciano-Cabral F, Cabral G (2003) Acanthamoeba sp. as agents of disease in humans. Clin Microbiol Rev 16:273–307Google Scholar
  17. Marciano-Cabral F, Puffenbauer R, Cabral GA (2000) The increasing importance of Acanthamoeba infections. J Eukaryot Microbiol 47:29–36Google Scholar
  18. Martínez A, Visvesvara G (1997) Free living, amphizoic and opportunistic amoebas. Brain Pathol 7:593–598Google Scholar
  19. Mergeryan H (1991) The prevalence of Acanthamoeba in the human environment. Rev Infect Dis 5:390–391Google Scholar
  20. Niederkorn JY, Alizadeh H, Leher H, McCulley JP (1999) The pathogenesis of Acanthamoeba keratitis. Microb Infect 1:437–443Google Scholar
  21. Ortega-Rivas A (2003) Aplicación de la técnica de RAPD para la identificación de especies del género Acanthamoeba. PhD thesis, University of La Laguna, Tenerife, ISBN 84-688-5110-5118Google Scholar
  22. Ortega-Rivas A, Lorenzo-Morales J, Alonso V, Abreu N, Foronda P, Del Castillo A Valladares B (2003) Random amplified polymorphic DNA as a tool for the identification of Acanthamoeba divionensis. Curr Microbiol 47:84–86Google Scholar
  23. Ortega-Rivas A, Lorenzo-Morales J, Villa M, Martinez E, Valladares B, Del Castillo A (2004) Design and evaluation of a specific primer pair for the identification of Acanthamoeba polyphaga. Curr Microbiol 48:360–363Google Scholar
  24. Parija SC, Prakash MR, Rao VA, Vellaniparambil RJ (2001) Acanthamoeba keratitis in Pondicherry. J Commun Dis 33:126–129Google Scholar
  25. Pussard M, Pons R (1977) Morphologie de la paroi kystique et taxonomie du genre Acanthamoeba. Protistologica 13:557–598Google Scholar
  26. Radford C, Lehman O, Dart JKG (1998) Acanthamoeba keratitis: multicentre survey in England 1992–6. National Acanthamoeba Keratitis Study Group. Br J Ophthalmol 82:1387–1392Google Scholar
  27. Rivera MA, Padhya TA (2002) Acanthamoeba: a rare primary cause of rhinosinusitis. Laryngoscope 112:1201–1203Google Scholar
  28. Rodríguez R, Méndez O, Molina O, Luzardo G, Martínez A, Visvesvara G, Cardozo J (1998) Infección del sistema nervioso central por amebas de vida libre: comunicación de tres nuevos casos venezolanos. Rev Neurol 26:1005–1008Google Scholar
  29. Sambrook J, Fristsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
  30. Seal D, Beattie TK, Tomlinson A, Fan D, Wong E (2003) Acanthamoeba keratitis. Br J Ophthalmol 87:516–517Google Scholar
  31. Seijo-Martínez M, González-Medeiro G, Santiago P, Rodríguez De Lope A, Diz J, Conde C, Visvesvara G (2000) Granulomatous amebic encephalitis in a patient with AIDS: isolation of Acanthamoeba sp. Group II from brain tissue and successful treatment with sulfadiazine and fluconazole. J Clin Microbiol 38:3892–3895Google Scholar
  32. Sell JJ, Rupp FW, Orrison WWJr (1997) Granulomatous amebic encephalitis caused by Acanthamoeba. Neuroradiology 39:434–436Google Scholar
  33. Sharma S, Garg P, Rao GN (2000) Patient characteristics, diagnosis, and treatment of non-contact lens related Acanthamoeba keratitis. Br J Ophthalmol 84:1103–1108Google Scholar
  34. Shasuzzaman SM, Hashiguchi Y (2002) Thoracic amoebiasis. Clin Chest Med 23:479–492Google Scholar
  35. Steinberg JP, Galindo RL, Kraus ES, Ghanem KG (2002) Disseminated acanthamoebiasis in a renal transplant recipient with osteomyelitis and cutaneous lesions: case report and literature review. Clin Infect Dis 35:43–49Google Scholar
  36. Stothard DR, Schroeder-Diedrich JM, Awwad MH, Gast RJ, Ledee DR, Rodríguez-Zaragoza S, Dean C, Fuerst PA, Byers TJ (1998) The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J Eukaryot Microbiol 45:45–54Google Scholar
  37. Szenasi Z, Endo T, Yagita K, Nagy E (1998) Isolation, identification and increasing importance of free-living amoebae, causing human disease. J Med Microbiol 47:45–54Google Scholar
  38. Vodkin MH, Howe DK, Visvesvara G, McLaughlin GL (1992) Identification of Acanthamoeba at the generic and specific levels using the polymerase chain reaction. J Protozool 39:378–385Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Jacob Lorenzo-Morales
    • 1
  • Antonio Ortega-Rivas
    • 1
    Email author
  • Pilar Foronda
    • 1
  • Enrique Martínez
    • 1
  • Basilio Valladares
    • 1
  1. 1.Department of ParasitologyUniversity of La LagunaLa LagunaSpain

Personalised recommendations