Parasitology Research

, Volume 116, Issue 4, pp 1273–1284 | Cite as

Update on Acanthamoeba jacobsi genotype T15, including full-length 18S rDNA molecular phylogeny

  • Daniele CorsaroEmail author
  • Martina Köhsler
  • Margherita Montalbano Di Filippo
  • Danielle Venditti
  • Rosa Monno
  • David Di Cave
  • Federica Berrilli
  • Julia Walochnik
Original Paper


Free-living amoebae of the genus Acanthamoeba are worldwide present in natural and artificial environments, and are also clinically important, as causative agents of diseases in humans and other animals. Acanthamoeba comprises several species, historically assigned to one of the three groups based on their cyst morphology, but presently recognized as at least 20 genotypes (T1-T20) on the basis of their nuclear 18S ribosomal RNA (rRNA) gene (18S rDNA) sequences. While strain identification may usually be achieved targeting short (<500 bp) 18S ribosomal DNA (rDNA) fragments, the use of full-length gene sequences (>2200 bp) is necessary for correct genotype description and reliable molecular phylogenetic inference. The genotype T15, corresponding to Acanthamoeba jacobsi, is the only genotype described on the basis of partial sequences (~1500 bp). While this feature does not prevent the correct identification of the strains, having only partial sequences renders the genotype T15 not completely defined and may furthermore affect its position in the Acanthamoeba molecular tree. Here, we complete this gap, by obtaining full-length 18S rDNA sequences from eight A. jacobsi strains, genotype T15. Morphologies and physiological features of isolated strains are reported. Molecular phylogeny based on full 18S rDNA confirms some previous suggestions for a genetic link between T15 and T13, T16, and T19, with T19 as sister-group to T15.


Acanthamoeba Acanthamoeba jacobsi Acanthamoeba T15 Group III acanthamoebae Group I intron 


  1. Alves JMP, Gusmão CX, Teixeira MM, Freitas D, Foronda AS, 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 33:19–26PubMedGoogle Scholar
  2. Armand B, Motazedian MH, Asgari Q (2016) Isolation and identification of pathogenic free-living amoeba from surface and tap water of Shiraz City using morphological and molecular methods. Parasitol Res 115:63–68CrossRefPubMedGoogle Scholar
  3. Booton GC, Kelly DJ, Chu Y, 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 40:1621–1625CrossRefPubMedPubMedCentralGoogle Scholar
  4. Booton GC, Visvesvara GS, Byers TJ, Kelly DJ, Fuerst PA (2005) Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. J Clin Microbiol 43:1689–1693CrossRefPubMedPubMedCentralGoogle Scholar
  5. Cabello-Vílchez AM, Martín-Navarro CM, López-Arencibia A, Reyes-Batlle M, González AC, Guerra H, Gotuzzo E, Valladares B, Piñero JE, Lorenzo-Morales J (2014) Genotyping of potentially pathogenic Acanthamoeba strains isolated from nasal swabs of healthy individuals in Peru. Acta Trop 130:7–10CrossRefPubMedGoogle Scholar
  6. Conza L, Pagani SC, Gaia V (2013) Presence of Legionella and free-living amoebae in composts and bioaerosols from composting facilities. PLoS One 8:e68244CrossRefPubMedPubMedCentralGoogle Scholar
  7. Corsaro D, Venditti D (2010) Phylogenetic evidence for a new genotype of Acanthamoeba (Amoebozoa, Acanthamoebida). Parasitol Res 107:233–238CrossRefPubMedGoogle Scholar
  8. Corsaro D, Venditti D (2011) More Acanthamoeba genotypes: limits to use rDNA fragments to describe new genotype. Acta Protozool 50:51–56Google Scholar
  9. Corsaro D, Walochnik J, Köhsler M, Rott MB (2015) Acanthamoeba misidentification and multiple labels: redefining genotypes T16, T19 and T20, and proposal for Acanthamoeba micheli sp. nov. (genotype T19). Parasitol Res 114:2481–2490CrossRefPubMedGoogle Scholar
  10. Curson RTM, Brown TJ (1978) Use of cell cultures as an indicator of pathogenicity of free-living amoebae. J Clin Pathol 31:1–11CrossRefGoogle Scholar
  11. Di Cave 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. Eur J Clin Microbiol Infect Dis 28:607–612CrossRefPubMedGoogle Scholar
  12. Di Cave D, D'Alfonso R, Dussey Comlavi KA, D'Orazi C, Monno R, Berrilli F (2014) Genotypic heterogeneity based on 18S-rRNA gene sequences among Acanthamoeba isolates from clinical samples in Italy. Exp Parasitol 145:S46–S49CrossRefPubMedGoogle Scholar
  13. Duarte JL, Furst C, Klisiowicz DR, Klassen G, Costa AO (2013) Morphological, genotypic, and physiological characterization of Acanthamoeba isolates from keratitis patients and the domestic environment in Vitoria, Espírito Santo, Brazil. Exp Parasitol 135:9–14CrossRefPubMedGoogle Scholar
  14. Evyapan G, Koltas IS, Eroglu F (2015) Genotyping of Acanthamoeba T15: the environmental strain in Turkey. Trans R Soc Trop Med Hyg 109:221–224CrossRefPubMedGoogle Scholar
  15. Fabres LF, Rosa dos Santos SP, Benitez LB, Rott MB (2016) Isolation and identification of Acanthamoeba spp. from thermal swimming pools and spas in Southern Brazil. Acta Parasitol 61:221–227CrossRefPubMedGoogle Scholar
  16. Flint JA, Dobson PJ, Robinson BS (2003) Genetic analysis of forty isolates of Acanthamoeba group III by multilocus isoenzyme electrophoresis. Acta Protozool 42:317–324Google Scholar
  17. Fritsche TR, Gautom RK, Seyerdirshti S, Bergeron DL, Lindquist TD (1993) Occurrence of bacterial endosymbionts in Acanthamoeba spp. isolated from corneal and environmental specimens and contact lenses. J Clin Microbiol 31:1122–1126PubMedPubMedCentralGoogle Scholar
  18. Gast RJ, Fuerst PA, Byers TJ (1994) Discovery of group I introns in the nuclear small subunit ribosomal RNA genes of Acanthamoeba. Nucleic Acids Res 22:592–596CrossRefPubMedPubMedCentralGoogle Scholar
  19. Geisen S, Fiore-Donno AM, Walochnik J, Bonkowski M (2014) Acanthamoeba everywhere: high diversity of Acanthamoeba in soils. Parasitol Res 113:3151–3158CrossRefPubMedGoogle Scholar
  20. Griffin JL (1972) Temperature tolerance of pathogenic and nonpathogenic free-living amoebae. Science 178:869–870CrossRefPubMedGoogle Scholar
  21. Grün AL, Stemplewitz B, Scheid P (2014) First report of an Acanthamoeba genotype T13 isolate as etiological agent of a keratitis in humans. Parasitol Res 113:2395–2400CrossRefPubMedGoogle Scholar
  22. Hajialilo E, Behnia M, Tarighi F, Niyyati M, Rezaeian M (2016) Isolation and genotyping of Acanthamoeba strains (T4, T9, and T11) from amoebic keratitis patients in Iran. Parasitol Res 115:3147–3151CrossRefPubMedGoogle Scholar
  23. Hedberg A, Johansen SD (2013) Nuclear group I introns in self-splicing and beyond. Mob DNA 4:17CrossRefPubMedPubMedCentralGoogle Scholar
  24. 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, Nerad and Visvesvara, 1992 (Lobosea: Acanthamoebidae). Acta Protozool 42:325–329Google Scholar
  25. Huang SW, Hsu BM (2010) Isolation and identification of Acanthamoeba from Taiwan spring recreation areas using culture enrichment combined with PCR. Acta Trop 115:282–287CrossRefPubMedGoogle Scholar
  26. Johansen S, Haugen P (2001) A new nomenclature of group I introns in ribosomal DNA. RNA 7:935–936CrossRefPubMedPubMedCentralGoogle Scholar
  27. Kao PM, Hsu BM, Chen NH, Huang KH, Huang SW, King KL, Chiu YC (2012) Isolation and identification of Acanthamoeba species from thermal spring environments in southern Taiwan. Exp Parasitol 130:354–358CrossRefPubMedGoogle Scholar
  28. Kao PM, Chou MY, Tao CW, Huang WC, Hsu BM, Shen SM, Fan CW, Chiu YC (2013) Diversity and seasonal impact of Acanthamoeba species in a subtropical rivershed. Biomed Res Int 2013:405794PubMedPubMedCentralGoogle Scholar
  29. Khan NA (2009) Acanthamoeba–biology and pathogenesis. Caister Academic Press, Norfolk, Great Britain, 290 ppGoogle Scholar
  30. Khan NA, Jarroll EL, Paget TA (2001) Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays. Curr Microbiol 43:204–208CrossRefPubMedGoogle Scholar
  31. Kiss C, Barna Z, Vargha M, Török JK (2014) Incidence and molecular diversity of Acanthamoeba species isolated from public baths in Hungary. Parasitol Res 113:2551–2557CrossRefPubMedGoogle Scholar
  32. Koltas IS, Eroglu F, Erdem E, Yagmur M, Tanır F (2015) The role of domestic tap water on Acanthamoeba keratitis in non-contact lens wearers and validation of laboratory methods. Parasitol Res 114:3283–3289CrossRefPubMedGoogle Scholar
  33. Landell MF, Salton J, Caumo K, Broetto L, Rott MB (2013) Isolation and genotyping of free-living environmental isolates of Acanthamoeba spp. from bromeliads in southern Brazil. Exp Parasitol 134:290–294CrossRefPubMedGoogle Scholar
  34. Maciver SK, Asif M, Simmen MW, Lorenzo-Morales J (2013) A systematic analysis of Acanthamoeba genotype frequency correlated with source and pathogenicity: T4 is confirmed as a pathogen-rich genotype. Eur J Protistol 49:217–221CrossRefPubMedGoogle Scholar
  35. Maghsood AH, Sissons J, Rezaian M, Nolder D, Warhurst D, Khan NA (2005) Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates. J Med Microbiol 54:755–759CrossRefPubMedGoogle Scholar
  36. Magnet A, Henriques-Gil N, Galván-Diaz AL, Izquiedo F, Fenoy S, Del Aguila C (2014) Novel Acanthamoeba 18S rRNA gene sequence type from an environmental isolate. Parasitol Res 113:2845–2850CrossRefPubMedGoogle Scholar
  37. Marciano-Cabral F, Cabral G (2003) Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 16:273–307CrossRefPubMedPubMedCentralGoogle Scholar
  38. Michel F, Westhof E (1990) Modelling of the three-dimensional architecture of group I catalytic introns based on comparative sequence analysis. J Mol Biol 216:585–610CrossRefPubMedGoogle Scholar
  39. Montalbano Di Filippo M, Santoro M, Lovreglio P, Monno R, Capolongo C, Calia C, Fumarola L, D'Alfonso R, Berrilli F, Di Cave D (2015) Isolation and molecular characterization of free-living amoebae from different water sources in Italy. Int J Environ Res Public Health 12:3417–3427CrossRefPubMedPubMedCentralGoogle Scholar
  40. Nagyová V, Nagy A, Janecek S, Timko J (2010a) Morphological, physiological, molecular and phylogenetic characterization of new environmental isolates of Acanthamoeba spp. from the region of Bratislava, Slovakia. Biologia 65:81–91CrossRefGoogle Scholar
  41. Nagyová V, Nagy A, Timko J (2010b) Morphological, physiological and molecular biological characterisation of isolates from first cases of Acanthamoeba keratitis in Slovakia. Parasitol Res 106:861–872CrossRefPubMedGoogle Scholar
  42. Niyyati M, Nazar M, Haghighi A, Nazemalhosseini Mojarad E (2012) Screening of recreational areas of rivers for potentially pathogenic free-living amoebae in the suburbs of Tehran, Iran. J Water Health 10:140–146CrossRefPubMedGoogle Scholar
  43. Niyyati M, Saberi R, Latifi A, Lasjerdi Z (2016) Distribution of Acanthamoeba genotypes isolated from recreational and therapeutic geothermal water sources in Southwestern Iran. Environ Health Insights 10:69–74CrossRefPubMedPubMedCentralGoogle Scholar
  44. Page FC (1988) A new key to freshwater and soil Gymnamoebae. Freshwater Biological Association, Ambleside, Cumbria, p 92–97Google Scholar
  45. Pussard M, Pons R (1977) Morphologie de la paroi kystique et taxonomie du genre Acanthamoeba (Protozoa, Amoebida). Protistologica 8:557–598Google Scholar
  46. Qvarnstrom Y, Nerad TA, Visvesvara GS (2013) Characterization of a new pathogenic Acanthamoeba species, A. byersi n. sp., isolated from a human with fatal amoebic encephalitis. J Eukaryot Microbiol 60:626–633CrossRefPubMedPubMedCentralGoogle Scholar
  47. Risler A, Coupat-Goutaland B, Pélandakis M (2013) Genotyping and phylogenetic analysis of Acanthamoeba isolates associated with keratitis. Parasitol Res 112:3807–3816CrossRefPubMedGoogle Scholar
  48. Rivera WL, Adao DE (2008) Identification of the 18S-ribosomal-DNA genotypes of Acanthamoeba isolates from the Philippines. Ann Trop Med Parasitol 102:671–677CrossRefPubMedGoogle Scholar
  49. Sawyer TK, Visvesvara GS, Harke BA (1977) Pathogenic amoebas from brackish and ocean sediments, with a description of Acanthamoeba hatchetti, n. sp. Science 196:1324–1325CrossRefPubMedGoogle Scholar
  50. Sawyer TK, Nerad TA, Visvesvara GS (1992) Acanthamoeba jacobsi sp. n. (Protozoa: Acanthamoebidae) from sewage contaminated ocean sediments. J Helminthol Soc Wash 59:223–226Google Scholar
  51. Schroeder JM, Booton GC, Hay J, Niszl IA, Seal DV, Markus MB, Fuerst PA, Byers TJ (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. J Clin Microbiol 39:1903–1911CrossRefPubMedPubMedCentralGoogle Scholar
  52. Schroeder-Diedrich JM, Fuerst PA, Byers TJ (1998) Group-I introns with unusual sequences occur at three sites in nuclear 18S rRNA genes of Acanthamoeba lenticulata. Curr Genet 34:71–78CrossRefPubMedGoogle Scholar
  53. Sharifi N, Botero-Kleiven S, Ohman D, Barragan A, Winiecka-Krusnell J (2010) Genotypic characterization of Acanthamoeba spp. causing ocular infections in Swedish patients: identification of the T15 genotype in a case of protracted keratitis. Scand J Infect Dis 42:781–786CrossRefPubMedGoogle Scholar
  54. Solgi R, Niyyati M, Haghighi A, Taghipour N, Tabaei SJ, Eftekhar M, Nazemalhosseini Mojarad E (2012) Thermotolerant Acanthamoeba spp. isolated from therapeutic hot springs in Northwestern Iran. J Water Health 10:650–656CrossRefPubMedGoogle Scholar
  55. 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 Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J Eukaryot Microbiol 45:45–54CrossRefPubMedGoogle Scholar
  56. Tanveer T, Hameed A, Gul A, Matin A (2015) Quick survey for detection, identification and characterization of Acanthamoeba genotypes from some selected soil and water samples in Pakistan. Ann Agric Environ Med 22:227–230CrossRefPubMedGoogle Scholar
  57. Tawfeek GM, Bishara SAH, Sarhan RM, Taher EE, Khayyal AE (2016) Genotypic, physiological, and biochemical characterization of potentially pathogenic Acanthamoeba isolated from the environment in Cairo, Egypt. Parasitol Res 115:1871–1881CrossRefPubMedGoogle Scholar
  58. Thong YH, Ferrante A (1986) Migration patterns of pathogenic and nonpathogenic Naegleria spp. Infect Immun 51:177–180PubMedPubMedCentralGoogle Scholar
  59. Tice AK, Shadwick LL, Fiore-Donno AM, Geisen S, Kang S, Schuler GA, Spiegel FW, Wilkinson KA, Bonkowski M, Dumack K, Lahr DJ, Voelcker E, Clauß S, Zhang J, Brown MW (2016) Expansion of the molecular and morphological diversity of Acanthamoebidae (Centramoebida, Amoebozoa) and identification of a novel life cycle type within the group. Biol Direct 11:69CrossRefPubMedPubMedCentralGoogle Scholar
  60. Walochnik J, Obwaller A, Aspöck H (2000) Correlation between morphological, molecular biological, and physiological characteristics in clinical and nonclinical isolates of Acanthamoeba spp. Appl Environ Microbiol 66:4408–4413CrossRefPubMedPubMedCentralGoogle Scholar
  61. Walochnik J, Michel R, Aspöck H (2004) A molecular biological approach to the phylogenetic position of the genus Hyperamoeba. J Eukaryot Microbiol 51:433–440CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.CHLAREAS Chlamydia Research AssociationVandoeuvre-lès-NancyFrance
  2. 2.Molecular Parasitology, Institute of Specific Prophylaxis and Tropical MedicineMedical University of ViennaViennaAustria
  3. 3.Department of Clinical Sciences and Translational MedicineUniversity of Rome “Tor Vergata”RomeItaly
  4. 4.TREDI Research Department, Faculty of MedecineVandoeuvre-lès-NancyFrance
  5. 5.Department of Basic Medical Science, Neuroscience and Sense OrgansUniversity of Bari “Aldo Moro”BariItaly

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