Advertisement

Parasitology Research

, Volume 105, Issue 4, pp 1109–1117 | Cite as

Isolation and genotyping of potentially pathogenic Acanthamoeba and Naegleria species from tap-water sources in Osaka, Japan

  • Akiko Edagawa
  • Akio KimuraEmail author
  • Takako Kawabuchi-Kurata
  • Yasuhiro Kusuhara
  • Panagiotis Karanis
Original Paper

Abstract

Here, we carried out a survey to determine the prevalence of free-living amoebae (FLA) in tap-water sources from rivers and water treatment plants located in Osaka Prefecture, Japan. A total of 374 raw water samples were collected from 113 sampling points. The samples were filtrated and transferred to non-nutrient agar plates seeded with a heat-killed suspension of Escherichia coli and incubated for 2 to 7 days at 30°C or 42°C. The plates were examined by microscopy to morphologically identify FLA families, and polymerase chain reaction and sequence analysis were then performed to define the species of the detected Naegleria and Acanthamoeba isolates. A total of 257 of 374 samples (68.7%) were positive for FLA by microscopy, and among these there were 800 FLA isolates, including Acanthamoeba and Naegleria species. Sequence analysis identified five Acanthamoeba spp. isolates of the known pathogenic T4 genotype and 43 Naegleria australiensis isolates, a reported pathogen to mice and also of concern as a potential pathogen to humans. Our results suggest a wide distribution of FLA, including potential pathogenic species, in tap-water sources of western Japan.

Keywords

Internal Transcribe Spacer Water Treatment Plant Osaka Prefecture Blast Homology Search Acanthamoeba Isolate 
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.

Notes

Acknowledgments

We thank Mr. H. Doi and Ms. T. Hizuka for technical assistance. We are grateful to Dr. T. Endo of the National Institute of Infectious Diseases, Japan for providing the DNA of N. lovaniensis. We are also grateful to Mr. B. Sheridan and Dr. G. Harris of DMC Corporation for their review of the manuscript during its preparation. This work was supported in part by grants to A. E from the Ministry of Education, Culture, Sports, Science, and Technology, Japan.

References

  1. Behets J, Declerck P, Delaedt Y, Verelst L, Ollevier F (2007) Survey for the presence of specific free-living amoebae in cooling waters from Belgian power plants. Parasitol Res 100:1249–1256PubMedCrossRefGoogle Scholar
  2. Chong EM, Dana MR (2007) Acanthamoeba keratitis. Int Ophthalmol Clin 47:33–46PubMedCrossRefGoogle Scholar
  3. De Jonckheere JF (2004) Molecular definition and the ubiquity of species in the genus Naegleria. Protist 155:89–103PubMedCrossRefGoogle Scholar
  4. De Jonckheere JF (2006) Isolation and molecular identification of free-living amoebae of the genus Naegleria from Arctic and sub-Antarctic regions. Europ J Protistol 42:115–123CrossRefGoogle Scholar
  5. De Jonckheere JF (2007) Molecular identification of free-living amoebae of the Vahlkampfiidae and Acanthamoebidae isolated in Arizona (USA). Europ J Protistol 43:9–15CrossRefGoogle Scholar
  6. De Jonckheere JF, Yagita K, Kuroki T, Endo T (1991) Furst isolation of pathogenic Naegrelia fowleri in Japan. Jpn J Parasitol 40:352–357Google Scholar
  7. 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 89:6–15PubMedCrossRefGoogle Scholar
  8. Gornik K, Kuzna-Grygiel W (2004) Presence of virulent strains of amphizoic amoebae in swimming pools of the city of Szczecin. Ann Agric Environ Med 11:233–236PubMedGoogle Scholar
  9. Greub G, Raoult D (2004) Microorganisms resistant to free-living amoebae. Clin Microbiol Rev 17:413–433PubMedCrossRefGoogle Scholar
  10. Hoffmann R, Michel R (2001) Distribution of free-living amoebae (FLA) during preparation and supply of drinking water. Int J Hyg Environ Health 203:215–219PubMedCrossRefGoogle Scholar
  11. Horn M, Wagner M (2004) Bacterial endosymbionts of free-living amoebae. J Eukaryot Microbiol 51:509–514PubMedCrossRefGoogle Scholar
  12. Huizinga HW, McLaughlin GL (1990) Thermal ecology of Naegleria fowleri from a power plant cooling reservoir. Appl Environ Microbiol 56:2200–2205PubMedGoogle Scholar
  13. Izumiyama S, Yagita K, Furushima-Shimogawara R, Asakura T, Karasudani T, Endo T (2003) Occurrence and distribution of Naegleria species in thermal waters in Japan. J Eukaryot Microbio 50:514–515CrossRefGoogle Scholar
  14. John DT, De Jonckheere JF (1985) Isolation of Naegleria australiensis from an Oklahoma Lake. J Protozool 32:571–575PubMedGoogle Scholar
  15. John DT, Howard MJ (1995) Seasonal distribution of pathogenic free-living amebae in Oklahoma waters. Parasitol Res 81:193–201PubMedGoogle Scholar
  16. Kandori M, Inoue T, Hori Y, Maeda N, Tano Y (2007) Clinical management and prognosis of Acanthamoeba keratitis: a retrospective study of 11 cases at the department of ophthalmology of Osaka university medical school. Folia Ophthalomol Jpn 58:691–636Google Scholar
  17. Karanis P, Kourenti C, Smith H (2007) Waterborne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt. J Water Health 5:1–38PubMedCrossRefGoogle Scholar
  18. Kilvington S, Gray T, Dart J, Morlet N, Beeching JR, Frazer DG, Matheson M (2004) Acanthamoeba keratitis: the role of domestic tap water contamination in the United Kingdom. Invest Ophthalomol Visual Sci 45:165–169CrossRefGoogle Scholar
  19. Ledee DR, Hay J, Byers TJ, Seal DV, Kirkness CM (1996) Acanthamoeba griffini. Molecular characterization of a new corneal pathogen. Invest Ophthalmol Visual Sci 37:544–550Google Scholar
  20. Ledee DR, Iovieno A, Miller D, Mandal N, Diaz M, Fell J, Fini ME, Alfonso EC (2009) Molecular identification of T4 and T5 genotypes in Acanthamoeba keratitis patients. J Clin Microbiol 47:1458–1462PubMedCrossRefGoogle Scholar
  21. Lorenzo-Morales J, Lindo JF, Martinez E, Calder D, Figueruelo E, Valladares B, Ortega-Rivas A (2005a) Pathogenic Acanthamoeba strains from water sources in Jamaica, West Indies. Ann Trop Med Parasitol 99:751–758PubMedCrossRefGoogle Scholar
  22. Lorenzo-Morales J, Ortega-Rivas A, Foronda P, Martinez E, Valladares B (2005b) Isolation and identification of pathogenic Acanthamoeba strains in Tenerife, Canary Islands, Spain from water sources. Parasitol Res 95:273–277PubMedCrossRefGoogle Scholar
  23. Lorenzo-Morales J, Ortega-Rivas A, Martinez E, Khoubbane M, Artigas P, Periago MV, Foronda P, Abreu-Acosta N, Valladares B, Mas-Coma S (2006) Acanthamoeba isolates belonging to T1, T2, T3, T4 and T7 genotypes from environmental freshwater samples in the Nile Delta region, Egypt. Acta Trop 100:63–69PubMedCrossRefGoogle Scholar
  24. Lorenzo-Morales J, Martinez-Carretero E, Batista N, Alvarez-Marin J, Bahaya Y, Walochnik J, Valladares B (2007) Early diagnosis of amoebic keratitis due to a mixed infection with Acanthamoeba and Hartmannella. Parasitol Res 102:167–169PubMedCrossRefGoogle Scholar
  25. Martinez AJ, Visvesvara GS (1997) Free-living, amphizoic and opportunistic amebas. Brain Pathol 7:583–598PubMedCrossRefGoogle Scholar
  26. Nacapunchai D, Kino H, Ruangsitticha C, Sriwichai P, Ishih A, Terada M (2001) A brief survey of free-living amebae in Thailand and Hamamatsu District Japan. Southeast Asian J Trop Med Pub Health 32(Suppl 2):179–182Google Scholar
  27. Page FC (1988) A new key to freshwater and soil Gymnamoebae. Ferry House, LondonGoogle Scholar
  28. Pelandakis M, Pernin P (2002) Use of multiplex PCR and PCR restriction enzyme analysis for detection and exploration of the variability in the free-living amoeba Naegleria in the environment. Appl Environ Microbiol 68:2061–2065PubMedCrossRefGoogle Scholar
  29. Scaglia M, Strosselli M, Grazioli V, Gatti S, Bernuzzi AM, de Jonckheere JF (1983) Isolation and identification of pathogenic Naegleria australiensis (Amoebida, Vahlkampfiidae) from a spa in northern Italy. Appl Environment Microbiol 46:1282–1285Google Scholar
  30. Scaglia M, Gatti S, Cevini C, Bernuzzi AM, Martinez AJ (1989) Naegleria australiensis ssp. italica: experimental study in mice. Exp Parasitol 69:294–299PubMedCrossRefGoogle Scholar
  31. Schroeder J, Booton GC, Hay J, Niszl IA, Seal DV, Markus MB, Fuerst PA, Byers TJ (2001) Use of subgenic 18 S ribosomal DNA PCR and sequencing for genus and genotype identification of Acanthamoebae from humans with keratitis and from sewage sludge. J Clin Microbiol 39:1903–1911PubMedCrossRefGoogle Scholar
  32. Schuster FL, Visvesvara GS (2004) Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals. Int J Parasitol 34:1001–1027PubMedCrossRefGoogle Scholar
  33. Seal DV (2003) Acanthamoeba keratitis update-incidence, molecular epidemiology and new drugs for treatment. Eye 17:893–905PubMedCrossRefGoogle Scholar
  34. Sheehan KB, Ferris MJ, Henson JM (2003) Detection of Naegleria sp. in a thermal, acidic stream in Yellowstone National Park. J Eukaryot Microbiol 50:263–265PubMedCrossRefGoogle Scholar
  35. Shoff ME, Rogerson A, Kessler K, Schatz S, Seal DV (2008) Prevalence of Acanthamoeba and other naked amoebae in South Florida domestic water. J Water Health 6:99–104PubMedCrossRefGoogle Scholar
  36. Spanakos G, Tzanetou K, Miltsakakis D, Patsoula E, Malamou-Lada E, Vakalis NC (2006) Genotyping of pathogenic Acanthamoebae isolated from clinical samples in Greece—report of a clinical isolate presenting T5 genotype. Parasitol Internat 55:147–149CrossRefGoogle Scholar
  37. Sugita Y, Fujii T, Hayashi I, Aoki T, Yokoyama T, Morimatsu M, Fukuma T, Takamiya Y (1999) Primary amebic meningoencephalitis due to Naegleria fowleri: an autopsy case in Japan. Pathol Internat 49:468–470CrossRefGoogle Scholar
  38. Thomas V, Loret JF, Jousset M, Greub G (2008) Biodiversity of amoebae and amoebae-resisting bacteria in a drinking water treatment plant. Environ Microbiol 10:2728–2745PubMedCrossRefGoogle Scholar
  39. Tsvetkova N, Schild M, Panaiotov S, Kurdova-Mintcheva R, Gottstein B, Walochnik J, Aspock H, Lucas MS, Muller N (2004) The identification of free-living environmental isolates of amoebae from Bulgaria. Parasitol Res 92:405–413PubMedCrossRefGoogle Scholar
  40. Visvesvara GS, De Jonckheere JF, Sriram R, Daft B (2005) Isolation and molecular typing of Naegleria fowleri from the brain of a cow that died of primary amebic meningoencephalitis. J Clin Microbiol 43:4203–4204PubMedCrossRefGoogle Scholar
  41. 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 50:1–26PubMedCrossRefGoogle Scholar
  42. Yamada T (2002) Advanced water treatment in Osaka. In: Osaka and Its Technology 41. Osaka Municipal Government, Osaka, pp 43–49Google Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Akiko Edagawa
    • 1
  • Akio Kimura
    • 2
    Email author
  • Takako Kawabuchi-Kurata
    • 3
  • Yasuhiro Kusuhara
    • 4
  • Panagiotis Karanis
    • 5
  1. 1.Division of Environmental HealthOsaka Prefectural Institute of Public HealthOsakaJapan
  2. 2.Division of Planning and CoordinationOsaka Prefectural Institute of Public HealthOsakaJapan
  3. 3.Division of VirologyOsaka Prefectural Institute of Public HealthOsakaJapan
  4. 4.Department of Medical Zoology, School of Health SciencesFujita Health UniversityToyoakeJapan
  5. 5.Medical and Molecular Parasitology, Centre of Anatomy, Institute II, Medical SchoolUniversity of CologneCologneGermany

Personalised recommendations