Original Article

Surveillance and Molecular Identification of Acanthamoeba and Naegleria Species in Two Swimming Pools in Alexandria University, Egypt


Background: Swimming in contaminated water was reported to be associated with Acanthamoeba and N. fowleri human infections. The present study was carried out with the aim of isolation and identification of the different species of Acanthamoeba and Naegleria from two swimming pools in Alexandria University.

Methods: Samples were collected from the swimming pools of Alexandria University Stadium and Faculty of Agriculture-Alexandria University during the period from May 2012 to April 2013.

Results: Free-living amoebae were prevalent in the collected samples. Molecular characterization confirmed the identity of ten Acanthamoeba isolates and seven Naegleria isolates. Acanthamoeba T3, T4, T5, T11 and T15 genotypes were identified. Acanthamoeba T4 was the most prevalent genotype.

Conclusion: The relatively high prevalence of Acanthamoeba, especially genotype T4, indicates the presence of a health hazard to swimmers particularly those wearing contact lenses. Naegleria fowleri was not found during the present study. 

Page FC. A new key to freshwater and soil gym-namoebae. Ambleside, UK: Freshwater Biological Association; 1988.

Araújo MFF, Medeiros MLQd, Neto LS, Amo-rim ADS. Spatial and temporal distribution of free-living protozoa in aquatic environments of a Brazilian semi-arid region. Rev Ambient Água. 2013;8(2):46-56.

da Rocha-Azevedo B, Tanowitz HB, Marciano-Cabral F. Diagnosis of infections caused by path-ogenic free-living amoebae. Interdiscip Perspect Infect Dis. 2009;2009:251406.

Qvarnstrom Y, da Silva AJ, Schuster FL et al. Molecular confirmation of Sappinia pedata as a causative agent of amoebic encephalitis. J Infect Dis. 2009;199(8):1139-42.

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

Marciano-Cabral F, Cabral G. Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev. 2003;16(2):273-307.

Shoff ME, Joslin CE, Tu EY et al. Efficacy of contact lens systems against recent clinical and tap water Acanthamoeba isolates. Cornea. 2008;27(6):713-9.

Schuster FL, Visvesvara GS. Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals. Int J Parasitol. 2004;34(9):1001-27.

Maruyama S, Matsuzaki M, Misawa K et al. Cya-nobacterial contribution to the genomes of the plastid-lacking protists. BMC Evol Biol. 2009;9:197.

Parija SC, Jayakeerthee SR. Naegleria fowleri: a free living amoeba of emerging medical importance. J Commun Dis. 1999;31(3):153-9.

Yoder JS, Eddy BA, Visvesvara GS et al. The epidemiology of primary amoebic meningoen-cephalitis in the USA, 1962-2008. Epidemiol Infect. 2010;138(7):968-75.

Ben Salah I, Drancourt M. Surviving within the amoebal exocyst: the Mycobacterium avium complex paradigm. BMC Microbiol. 2010;10:99.

Scheid P, Schwarzenberger R. Acanthamoeba spp. as vehicle and reservoir of adenoviruses. Parasitol Res. 2012;111(1):479-85.

Garcia A, Goni P, Cieloszyk J et al. Identification of free-living amoebae and amoeba-associated bacteria from reservoirs and water treatment plants by molecular techniques. Environ Sci Technol. 2013;47(7):3132-40.

Scheid P. Relevance of free-living amoebae as hosts for phylogenetically diverse microorganisms. Parasitol Res. 2014;113(7):2407-14.

Donati M, Cremonini E, Di Francesco A et al. Prevalence of Simkania negevensis in chlorinated water from spa swimming pools and domestic supplies. J Appl Microbiol. 2015;118(4):1076-82.

Cirillo JD, Falkow S, Tompkins LS et al. Interaction of Mycobacterium avium with environmental amoebae enhances virulence. Infect Immun. 1997;65(9):3759-67.

Greub G, Raoult D. Microorganisms resistant to free-living amoebae. Clin Microbiol Rev. 2004;17(2):413-33.

Thomas V, McDonnell G, Denyer SP et al. Free-living amoebae and their intracellular pathogenic microorganisms: risks for water quality. FEMS Microbiol Rev. 2010;34(3):231-59.

Al-Herrawy A, Bahgat M, Mohammed AE et al. Acanthamoeba species in swimming pools of Cairo, Egypt. Iran J Parasitol. 2014;9(2):194-201.

Al-Herrawy AZ, Heshmat MG, Abu Kabsha SH, Gad MA, Lotfy WM. Occurrence of Acanthamoeba species in the Damanhour Drinking Water Treatment Plant, Behera Governorate (Egypt). Rep Parasitol. 2015;4:15-21.

AI-Herrawy AZ, Mohamed SH, Medhat A, Mo-hammed AH, Gad MA. Distribution of Naegleria in water resources in Egypt. Egypt J Environ Res. 2014;2:1-14.

Hikal WM, Al-Herrawy AZ, Bahgat MM, Mo-hammed AH, Ashour AA. Detection of Naegleria isolates from the Egyptian aquatic environment. Public Health Prev Med. 2015;1(2):73-7.

Al-Herrawy AZ, Gad MA. Isolation and molecular identification of Naegleria fowleri from Nile River, Egypt. J Egypt Public Health Assoc. 2015;90(4):161-5.

Martinez AJ. Is Acanthamoeba encephalitis an oppor-tunistic infection? Neurology. 1980;30(6):567-74.

Schuster FL, Visvesvara GS. Opportunistic amoebae: challenges in prophylaxis and treatment. Drug Resist Updat. 2004;7(1):41-51.

Page FC. Taxonomic criteria for limax amoebae, with descriptions of 3 new species of Hartmannella and 3 of Vahlkampfia. J Protozool. 1967;14(3):499-521.

Page FC. An illustrated key to freshwater and soil amoebae. Scientific publication no. 34. Kendal, UK: Freshwater Biological Association; 1976.

Sheehan DC, Harpchak BB. Theory and practice of histotechnology. 2nd ed. London: The C.V. Mosby Company; 1980.

Winnepenninckx B, Backelijau T, De wachter R. Extraction of high molecular weight DNA from mollusca. Trends Genet. 1993;9(12):407.

Qvarnstrom Y, Visvesvara GS, Sriram R et al. Multiplex Real-Time PCR Assay for simultaneous detection of Acanthamoeba spp., Balamuthia mandrillaris, and Naegleria fowleri. J Clin Microbiol. 2006;44(10):3589-95.

Schild M, Gianinazzi C, Gottstein B et al. PCR-based diagnosis of Naegleria sp. infection in forma-lin-fixed and paraffin-embedded brain sections. J Clin Microbiol. 2007;45(2):564-7.

Pelandakis M, Serre S, Pernin P. Analysis of the 5.8S rRNA gene and the internal transcribed spacers in Naegleria spp. and in N. fowleri. J Eukaryot Microbiol. 2000;47(2):116-21.

Bonadonna L, Briancesco R, Paradiso R, Semproni M. Free-living amoebae and enteric protozoa isolated in swimming pool. In: Ferretti E, Fantuzzi G, Spica VR, Caroli S, Bonadonna L, editors. The 5th International Conference Swim-ming Pool and Spa; lstituto Superiore di Sanita and Universita di Roma Faro ltalico Rome, April 9-122013.

Abd El-Salam MM. Assessment of water quality of some swimming pools: A case study in Alexandria, Egypt. Environ Monit Assess. 2012;184(12):7395-406.

Init I, Lau YL, Arin Fadzlun A et al. Detection of free living amoebae, Acanthamoeba and Naegleria, in swimming pools, Malaysia. Trop Biomed. 2010;27(3):566-77.

Tzeng KJ, Tung MC, Hsu BM et al. Detection and identification of free-living amoeba from aquatic environment in Taiwan. EGU General Assembly; Vienna, Austria, 7-12 April2013.

Mazur T. Occurrence of Naegleria fowleri in a free environment and biological properties of isolated strains. Wiad Parazytol. 1984;30(1):3-35.

Mazur T, Hadas E, Iwanicka I. The duration of the cyst stage and the viability and virulence of Acanthamoeba isolates. Trop Med Parasitol. 1995;46(2):106-8.

De Jonckheere J, van de Voorde H. Differences in destruction of cysts of pathogenic and non-pathogenic Naegleria and Acanthamoeba by chlorine. Appl Environ Microbiol. 1976;31(2):294-7.

Sarkar P, Gerba CP. Inactivation of Naegleria Fowleri by chlorine and ultraviolet light. J Am Water Works Assoc. 2012;104:51-2.

Tyndall RL, Ironside KS, Metler PL et al. Effect of thermal additions on the density and distribution of thermophilic amoebae and pathogenic Naegleria fowleri in a newly created cooling lake. Appl Environ Microbiol. 1989;55(3):722-32.

Moussa M, De Jonckheere JF, Guerlotte J et al. Survey of Naegleria fowleri in geothermal recreational waters of Guadeloupe (French West Indies). PLoS One. 2013;8(1):e54414.

Nagyova V, Nagy A, Janecek S, Timko J. Morpho-logical, physiological, molecular and phylogenetic characterization of new environmental isolates of Acanthamoeba spp. from the region of Bratislava, Slovakia. Biologia (Bratisl). 2010;65(1):81–91.

Martinez AJ. Free-living amebas: infection of the central nervous system. Mt Sinai J Med. 1993;60(4):271-8.

Booton GC, Visvesvara GS, Byers TJ et al. Iden-tification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. J Clin Microbiol. 2005;43(4):1689-93.

Alsam S, Kim KS, Stins M et al. Acanthamoeba interactions with human brain microvascular en-dothelial cells. Microb Pathog. 2003;35(6):235-41.

Crary MJ. Genetic variability and its relationship to Acanthamoeba pathogenesis. Ph.D. Thesis. Graduate School, Ohio State University. Ohio, USA 2012.

Ledee DR, Hay J, Byers TJ, Seal DV, Kirkness CM. Acanthamoeba griffini Molecular characterization of a new corneal pathogen. Invest Ophthalmol Vis Sci. 1996;37:544-50.

Barete S, Combes A, de Jonckheere JF et al. Fatal disseminated Acanthamoeba lenticulata infection in a heart transplant patient. Emerg Infect Dis. 2007;13(5):736-8.

Booton GC, Rogerson A, Bonilla TD et al. Mo-lecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis. J Eukaryot Microbiol. 2004;51(2):192-200.

IssueVol 12 No 2 (2017) QRcode
SectionOriginal Article(s)
Swimming water Acanthamoeba Naegleria Identification PCR

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How to Cite
Z. AL-HERRAWY A, I. KHALIL M, S. EL-SHERIF S, A. E. OMAR F, M. LOTFY W. Surveillance and Molecular Identification of Acanthamoeba and Naegleria Species in Two Swimming Pools in Alexandria University, Egypt. Iran J Parasitol. 2017;12(2):196-205.