Parasitology Research

, Volume 111, Issue 5, pp 2017–2022 | Cite as

Prevalence of Acanthamoeba spp. (Sarcomastigophora: Acanthamoebidae) in wild populations of Aedes aegypti (Diptera: Culicidae)

  • Dayane Andriotti Otta
  • Marilise Brittes Rott
  • Ana Maris Carlesso
  • Onilda Santos da SilvaEmail author
Original Paper


Studies of interrelationship between microorganisms and mosquitoes are of great importance, since it can provide support for better understand related to biology, development and their control. In this way, it is known that mosquito larvae and free-living amoebae (FLA) normally occupy similar aquatic microhabitats. However, few studies have been conducted about such coexistence. For that reason, the objective of the present study was to verify the prevalence of Acanthamoeba spp. in wild populations of Aedes aegypti, as well as to characterize the genotypic lineage, and their possible pathogenicity through thermo- and osmotolerance. Amoebae were investigated in 60 pools, each containing ten larvae of A. aegypti, collected in Porto Alegre (Rio Grande do Sul, Brazil). The Acanthamoeba isolates were morphologically characterized and submitted to the polymerase chain reaction technique to confirm identification of the genus. In addition, genotype analyses as well as tests for presumptive pathogenicity in some samples were performed. Of the 60 pools examined, 54 (90 %) were positive for FLA. Of these isolates, 47 (87 %) belonged to the genus Acanthamoeba. The genotypic groups T4, T3 and T5 were identified, numbering 14 (53.8 %), ten (38.5 %) and two (7.7 %) isolates, respectively. The physiological tests performed with 14 strains showed that 12 (85.7 %) were non-pathogenic, while two (14.3 %) were considered as having low pathogenic potential. These results provide a basis for a better understanding of the interaction between these protozoan and mosquitoes in their natural habitat. This study is the first to report the isolation of Acanthamoeba spp. from wild mosquitoes.


Wild Population Dengue Virus Mosquito Larva Cytoplasmatic Incompatibility Brazilian Amazon Region 
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.



The authors thank the National Council of Research and Development (CNPq/478119/2010-0) for financial support and Coordination for the Improvement of Higher Level Personal (Capes) for scholarship. Also, we thank Dr. Jaime Figueroa, Dr. Denise Haussmann and MSc. Adolfo Isla by contributions of this work in molecular biology.


  1. Albicócco AP, Vezzani D (2009) Further study on Ascogregarina culicis in temperate Argentina: prevalence and intensity in Aedes aegypti larvae and pupae. J Invertebr Pathol 101:210–214PubMedCrossRefGoogle Scholar
  2. Aljanabi SM, Martinez I (1997) Universal and rapid salt extraction of high quality genomic DNA for PCR based techniques. Nucleic Acids Res 25:4692–4693PubMedCrossRefGoogle Scholar
  3. Barker J, Brown MR (1994) Trojan horses of the microbial world: protozoa and the survival of bacterial pathogens in the environment. Microbiology 140:1253–1259PubMedCrossRefGoogle Scholar
  4. Bian G, Xu Y, Lu P, Xie Y, Xi Z (2010) The endosymbiotic bacterium Wolbachia induces resistance to dengue virus in Aedes aegypti. PLoS Pathog 6:e1000833PubMedCrossRefGoogle Scholar
  5. Booton GC, Rogerson A, Bonilla TD, Seal DV, Kelly DJ, Beattie TK, Tomlinson A, Lares-Villa F, Fuerst PA, Byers TJ (2004) Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis. J Eukaryot Microbiol 51:192–200PubMedCrossRefGoogle Scholar
  6. Brückner D, Garcia LS (1993) Diagnostic medical parasitology, 2nd edn. American Society for Microbiology, Washington, D.CGoogle Scholar
  7. Carlesso AM, Artuso GL, Caumo K, Rott MB (2010) Potentially Pathogenic Acanthamoeba Isolated from a Hospital in Brazil. Curr Microbiol 60:185–190PubMedCrossRefGoogle Scholar
  8. Caumo K, Rott MB (2011) Acanthamoeba T3, T4 and T5 in swimming-pool waters from Southern Brazil. Acta Trop 117:233–235PubMedCrossRefGoogle Scholar
  9. Chen WJ (1999) The life cycle of Ascogregarina taiwanensis (Apicomplexa: Lecudinidae). Parasitol Today 15:153–156PubMedCrossRefGoogle Scholar
  10. Declerck P, Behets J, van Hoef V, Ollevier F (2007) Detection of Legionella spp. and some of their amoeba hosts in floating biofilms from anthropogenic and natural aquatic environments. Water Res 41:3159–3167PubMedCrossRefGoogle Scholar
  11. Dellapé ME, Marti GA, Tranchida MC, García JJ (2005) First record of Aedes aegypti (L.) (Diptera: Culicidae) infected by the parasite Ascogregarina culicis (Ross) (Apicomplexa: Lecudinidae) in Argentina. Entomol Vect 12:111–115Google Scholar
  12. Dos Passos RA, Tadei WP (2008) Parasitism of Ascogregarina taiwanensis and Ascogregarina culicis (Apicomplexa: Lecudinidae) in larvae of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) from Manaus, Amazon region, Brazil. J Invertebr Pathol 97:230–236PubMedCrossRefGoogle Scholar
  13. 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. Parasitol Res 105:1109–1117PubMedCrossRefGoogle Scholar
  14. Fellous S, Koella JC (2009) Infectious dose affects the outcome of the within-host competition between parasites. Am Nat 173:E177–E184PubMedCrossRefGoogle Scholar
  15. Gaio Ade O, Gusmão DS, Santos AV, Berbert-Molina MA, Pimenta PF, Lemos FJ (2011) Contribution of midgut bacteria to blood digestion and egg production in Aedes aegypti (diptera: culicidae) (L.). Parasit Vectors 4:105PubMedCrossRefGoogle Scholar
  16. García-Munguía AM, Garza-Hernández JA, Rebollar-Tellez EA, Rodríguez-Pérez MA, Reyes-Villanueva F (2011) Transmission of Beauveria bassiana from male to female Aedes aegypti mosquitoes. Parasit Vectors 4:24PubMedCrossRefGoogle Scholar
  17. Gianinazzi C, Schild M, Zumkehr B et al (2010) Screening of Swiss hot spring resorts for potentially pathogenic free-living amoebae. Exp Parasitol 126:45–53PubMedCrossRefGoogle Scholar
  18. Kawaguchi K, Matsuo J, Osaki T, Kamiya S, Yamaguchi H (2009) Prevalence of Helicobacter and Acanthamoeba in natural environment. Lett Appl Microbiol 48:465–471PubMedCrossRefGoogle Scholar
  19. Khan NA, Jarroll EL, Paget T (2001) Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays. Curr Microbiol 43:204–208PubMedCrossRefGoogle Scholar
  20. Khan NA, Tareen NK (2003) Genotypic, phenotypic, biochemical, physiological and pathogenicity-based categorization of Acanthamoeba strains. Folia Parasitol 50:97–104PubMedGoogle Scholar
  21. Kingston D, Warhurst DC (1969) Isolation of amoebae from the air. J Med Microbiol 2:27–36PubMedCrossRefGoogle Scholar
  22. Koehsler M, Leitsch D, Duchêne M, Nagl M, Walochnik J (2009) Acanthamoeba castellanii: growth on human cell layers reactivates attenuated properties after prolonged axenic culture. FEMS Microbiol Lett 299:121–127PubMedCrossRefGoogle Scholar
  23. Lorenzo-Morales J, Ortega-Rivas A, Foronda P, Martínez E, Valladares B (2005) Isolation and identification of pathogenic Acanthamoeba strains in Tenerife, Canary Islands, Spain from water sources. Parasitol Res 95:273–277PubMedCrossRefGoogle Scholar
  24. Massad E, Coutinho FA (2011) The cost of dengue control. Lancet 377:1630–1631PubMedCrossRefGoogle Scholar
  25. Mortazavi PN, Goldsworthy G, Kirk R, Khan NA (2010) Acanthamoeba produces disseminated infection in locusts and traverses the locust blood–brain barrier to invade the central nervous system. BMC Microbiol 10:186PubMedCrossRefGoogle Scholar
  26. Niyyati M, Lorenzo-Morales J, Rahimi F, Motevalli-Haghi A, Martín-Navarro CM, Farnia S, Valladares B, Rezaeian M (2009) Isolation and genotyping of potentially pathogenic Acanthamoeba strains from dust sources in Iran. Trans R Soc Trop Med Hyg 103:425–427PubMedCrossRefGoogle Scholar
  27. Page FC (1988) A new key to freshwater and soil Gymnamoebae with instructions for culture. Freshwater Biological Association, CumbriaGoogle Scholar
  28. Paterson GN, Rittig M, Siddiqui R, Khan NA (2011) Is Acanthamoeba pathogenicity associated with intracellular bacteria? Exp Parasitol 129:207–210PubMedCrossRefGoogle Scholar
  29. Pussard M, Pons R (1977) Morphologie de la paroi kystique et taxonomie du genre Acanthamoeba (Protozoa, Amoebida). Protistologica 13:557–598Google Scholar
  30. Reyes-Villanueva F, Becnel JJ, Butler JF (2003) Susceptibility of Aedes aegypti and Aedes albopictus larvae to Ascogregarina culicis and Ascogregarina taiwanensis (Apicomplexa: Lecudinidae) from Florida. J Invertebr Pathol 84:47–53PubMedCrossRefGoogle Scholar
  31. Rott M, Caumo K, Sauter I, Eckert J, da Rosa L, da Silva O (2010) Susceptibility of Aedes aegypti (Diptera: Culicidae) to Acanthamoeba polyphaga (Sarcomastigophora: Acanthamoebidae). Parasitol Res 107:195–198PubMedCrossRefGoogle Scholar
  32. Sawyer TK (1971) Acanthamoeba griffini, a new species of marine amoeba. J Protozool 18:650–654Google Scholar
  33. Scheid PL, Schwarzenberger R (2011) Free-living amoebae as vectors of cryptosporidia. Parasitol Res 109:499–504PubMedCrossRefGoogle Scholar
  34. Scholte EJ, Takken W, Knols BG (2007) Infection of adult Aedes aegypti and Ae. albopictus mosquitoes with the entomopathogenic fungus Metarhizium anisopliae. Acta Trop 102:151–158PubMedCrossRefGoogle Scholar
  35. Schroeder JM, 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 Acanthamoeba from humans with keratitis and from sewage sludge. J Clin Microbiol 39:1903–1911PubMedCrossRefGoogle Scholar
  36. Siddiqui R, Emes R, Elsheikha H, Khan NA (2011) Area 51: How do Acanthamoeba invade the central nervous system? Trends Parasitol 27:185–189PubMedCrossRefGoogle Scholar
  37. Stockman LJ, Wright CJ, Visvesvara GS, Fields BS, Beach MJ (2011) Prevalence of Acanthamoeba spp. and other free-living amoebae in household water, Ohio, USA—1990–1992. Parasitol Res 108:621–627PubMedCrossRefGoogle Scholar
  38. 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 18 s rRNA gene sequence types. J Eukaryot Microbiol 45:45–54PubMedCrossRefGoogle Scholar
  39. Stratford MP, Griffiths AJ (1978) Variations in the properties and morphology of cysts of Acanthamoeba castellanii. J Gen Microbiol 108:33–37CrossRefGoogle Scholar
  40. 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
  41. Vasilakis N, Weaver SC (2008) The history and evolution of human dengue emergence. Adv Virus Res 72:1–76PubMedCrossRefGoogle Scholar
  42. Vezzani D, Wisnivesky C (2006) Prevalence and seasonality of Ascogregarina culicis (Apicomplexa: Lecudinidae) in natural populations of Aedes aegypti (Diptera: Culicidae) from temperate Argentina. J Invertebr Pathol 91:183–187PubMedCrossRefGoogle Scholar
  43. Visvesvara GS (1991) Classification of Acanthamoeba. Rev Infect Dis 13:369–372CrossRefGoogle Scholar
  44. Weiss B, Aksoy S (2011) Microbiome influences on insect host vector competence. Trends Parasitol 27:514–522PubMedCrossRefGoogle Scholar
  45. Winck MAT, Caumo K, Rott MB (2011) Prevalence of Acanthamoeba from tap water in Rio Grande do Sul, Brazil. Curr Microbiol 63:464–469PubMedCrossRefGoogle Scholar
  46. Zouache K, Raharimalala FN, Raquin V, Tran-Van V, Raveloson LH, Ravelonandro P, Mavingui P (2011) Bacterial diversity of field-caught mosquitoes, Aedes albopictus and Aedes aegypti, from different geographic regions of Madagascar. FEMS Microbiol Ecol 75:377–389PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Dayane Andriotti Otta
    • 1
  • Marilise Brittes Rott
    • 1
  • Ana Maris Carlesso
    • 1
  • Onilda Santos da Silva
    • 1
    Email author
  1. 1.Departamento de Microbiologia, Imunologia e Parasitologia, Instituto de Ciências Básicas da Saúde, Setor de ParasitologiaUniversidade Federal do Rio Grande do SulPorto AlegreBrazil

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