Iranian Journal of Parasitology 2014. 9(2):194-201.

Acanthamoeba species in Swimming Pools of Cairo, Egypt.
Ahmad Al-Herrawy, Mahmoud Bahgat, Abd-Elhafez Mohammed, Ameen Ashour, Wafaa Hikal


Background: The free-living amoebae Acanthamoeba spp. have been recognized as etiologic agents of amoebic encephalitis, keratitis, otitis, lung lesions and other skin infections mainly in immuno-compromised individuals. The purpose of this study is to detect the presence of Acanthamoeba in swimming pools in Egypt using a polymerase chain reaction (PCR) method.

Methods: Water samples were collected from 10 different swimming pools in Cai-ro, Egypt. Samples were cultured on non-nutrient agar for the detection of Acan-thamoeba isolates that were confirmed by PCR amplification using genus specific primers. The molecularly confirmed Acanthamoeba isolates were morphologically identified to the species level.

Results: Members of genus Acanthamoeba were detected in 49.2% of the examined swimming-pool water samples. Morphologically, six Acanthamoeba species were iso-lated from the examined swimming pool water namely A. polyphaga, A.castellanii, A. rhysodes, A. mauritaniensis, A. royreba and A. triangularis. All the identified species of Acanthamoeba were molecularly confirmed to be related to the genus Acanthamoeba.

Conclusion: The isolated species of Acanthamoeba could provoke variable degrees of infections to the swimmers. The culture method is cheaper and easier than PCR techniques that are faster for the detection of free-living amoebae.


Acanthamoeba; Cyst morphology; Egypt; PCR; Swimming pools

Full Text:



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

Marciano-Cabral F, Cabral GA. Acanthamoeba spp. as agents of disease in humans. Clin Mi-crobiol Rev. 2003; 16:273–307.

Marciano-Cabral F, Puffenbarger R, Cabral GA. The increasing importance of Acan-thamoeba infections. J Eukaryot Microbiol.2000; 47:29–36.

Martinez AJ, Visvesvara GS. Free-living am-phizoic and opportunistic amebas. Brain Pathol.1997; 7:583–598.

Moura H, Wallace S, Visvesvara.GS. Acan-thamoeba healyin.sp.and the isozyme and im-munoblot profiles of Acanthamoeba spp., groups 1 and 3. J Protozool.1992; 39:573–583.

Booton GC, Visvesvara GS., Byers TJ, Kelly DJ, Fuerst PA. Identification and Distribution of Acanthamoeba Species Genotypes Associated with Non-keratitis Infections. J Clin Microbiol. 2005; 43: 1689–1693.

Michel R, Muller KD, Amann R, Schmid EN. Legionella-like slender rods multiplying within a strain of Acanthamoeba spp. isolated from drink-ing water. Parasitol Res. 1998; 84: 84-88.

Barbaree JM, Fields BS, Feeley JC, Gorman GW, Martin WT. Isolation of protozoa from water associated with a legionellosis outbreak and demonstration of intracellular multiplica-tion of Legionella pneumophila. Appl Environ Mi-crobiol. 1986; 51: 422-424.

Rivera F, Ramirez P, Vilaclara G, Robles E, Medina F. A survey of pathogenic and free-liv-ing amoebae inhabiting swimming pool water in Mexico City. Environ Res.1983; 32: 205-211.

Scaglia M, Strosselli M, Grazioli V, Gatti S, BernuzziAM,DeJonckheere JF. Isolation and identification of pathogenic Naegleria australiensis (Amoebida, Vahlkampfidae) from spa in north-ern Italy. App Environ Microbiol.1983; 46: 1282-1285.

Thomas V, Herrera-Rimann K, Blanc DS, Greub G. Biodiversity of amoebae and amoe-bae-resisting bacteria in a hospital water net-work. Appl Environ Microbiol.2006; 72: 2428-2438.

Karanis P, Kourenti C, Smith H. Waterborne transmission of protozoan parasites: A world-wide review of outbreaks and lessons learnt. J Wat Health.2007; 5: 1-38.

Lehmann OJ, Green SM, Morlet N, Kilvington S, Keys MF, Matheson MM, Dart JK, McGill JI, Watt PJ. Polymerase chain reaction analysis of corneal epithelial and tear samples in the di-agnosis of Acanthamoeba keratitis. Invest Oph-thalmol Vis Sci. 1998; 39:1261–1265.

Houang E, Lam D, Fan D, Seal D. Microbial keratitis in Hong Kong: relationship to climate, environment and contact-lens disinfection. Trans R Soc Trop Med Hyg. 2001;95:361–367.

Booton GC, Kelly DJ, Chu YW, Seal DV, Houang E, Lam DSC, Byers TJ, Fuerst PA. 18S ribosomal DNA typing and tracking of Acanthamoeba species isolates from corneal scrape specimens, contact lenses, lens cases, and home water supplies of Acanthamoeba kera-titis patients in Hong Kong. J Clin Microbiol. 2002; 40:1621–1625.

Booton GC, Rogerson A, Bonilla TD, Seal DV, Kelly DJ, Beattie TK, Tomlinson A, Lares-Villa F, Fuerst PA, Byers TJ. Molecular and physiological evaluation of subtropical environ-mental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis. J Eukaryot Mi-crobiol. 2004; 51:192–200.

Pussard M, Pons R. Morphologie de la paroikystique et taxonomie du genre Acan-thamoeba (Protozoa, Amoebida). Protistol. 1977; TXIII: 557-598.

De Jonckheere J, Van Dijk P, De Voord H. Evaluation of the indirect fluorescent antibody technique for identification of Naegleria species. Appl Microbiol.1974; 28: 159-164.

Page FC. Nackte Rhizopoda, p. 3–145. In: D. Matthes (ed.), Protozoen fauna, Band 2. G. Fischer, Stuttgart, Germany.1991.

Winnepenninckx B, Backelijau T, de Wachter R. Extraction of high molecular weight DNA from mollusca. Trends Gen. 1993; 9: 407.

Abdel-Hamid AZ, Molfetta JB, Fernandez V, Rodrigues V. Genetic variation between sus-ceptible and non-susceptible snails to Schisto-soma infection using random amplified poly-morphic DNA analysis (RAPDs). Rev Inst Med Trop. 1999; 41: 291-295.

Walochnik J, Obwaller A, Aspock H. Correla-tion between morphological, molecular, biolog-ical and physiological characteristics in clinical and nonclinical isolates of Acanthamoeba spp. Appl Environ Microbiol. 2000; 66: 4408-4413.

Kilic A, Tanyuksel M, Sissons J, Jayasekera S, Khan N. Isolation of Acanthamoeba isolates be-longing to T2, T3, T4, and T7 genotypes from environmental samples in Ankara, Turkey. Ac-ta Parasitol. 2004; 49: 246–252.

Helling RB, Goodman HM Boyer HW. Analy-sis of R. EcoRI fragments of DNA from lamb-doid bacteriophages and other viruses by aga-rose-gel electrophoresis. J Virol. 1974; 14: 1235-38.

Al-Herrawy AZ. In vitro cultivation of agents of amoebic meningo-encephalitis isolated from water and sewage. Ph.D. thesis, Fac. Vet. Med., Alexandria Univ., Egypt. 1992.

Page F. A New Key to Freshwater and Soil Gymnamoebae. Freshwater Biol. Ass: Amble-side,1988.

Al-Herrawy AZ, Bahgat M, Mohammed A, Ashour A, Hikal W. Morpho-physiological and biochemical criteria of Acanthamoeba spp. iso-lated from the Egyptian aquatic environment. Iranian J Parasitol.2013; 8: 302-312.

Hilali M, Ashmawy K, Samaha H, Draz A, Abu El-Wafa S, Salem A. Preliminary studies on amoebic pathogens isolated from water and sewage with respect to Naegleria and Acan-thamoeba. J Egypt Vet Med Ass. 1994; 53: 215-224.

Ashmawy K, Hilali M, Abu El-Wafa SA, Sa-maha H, Draz AA, Salem A. In vitro identifica-tion of Naegleria and Acanthamoeba isolated from water and sewage. Assiut Vet Med J. 1993; 30: 87-100.

Gronik K, Kuzna-Grygiel W. Presence of viru-lent strains of amphizoic amoebae in swim-ming pools of the city of Szczecin. Ann Agric Environ Med. 2004; 11: 233-236.

Page FC. An Illustrated Key to Freshwater and Soil Amoebae. Freshwater Biol. Ass. 1976; ISSN 0367-I8887.SBN 900386266.

Chang SL. Etiological, pathological, epidemio-logical and diagnostical considerations of pri-mary amoebic meningo-encephalitis.Crit Rev Microbiol.1974; 3: 135-159.

Schuster FL, Visvesvara GS. Opportunistic amoebae: challenges in prophylaxis and treat-ment. Drug Resistance Updates.2004; 7:41-51.

Swofford D. Phylogenetic analysis using parsi-mony and other methods, PAUP*4.0b. Sun-derland, MA, USA: Sinauer Associates.1998.

Khan NA, Paget TA. Molecular tools for spe-ciation and epidemiological studies of Acan-thamoeba. Curr Microbiol. 2002; 44: 444-449.

Lorenzo-Morals J, Ortiga-Rivas A, Martinez E, Khoubbane M, Artigas P, Periago MV, Foron-da P, Abreu-Acosta N, Valladares B, Mas-Coma S. Acanthamoeba isolates belonging to T1, T2, T3, T4 and T7 genotypes from envi-

ronmental freshwater samples in the Nile Delta region, Egypt. Acta Trop. 2006; 100: 63-69.

Maghsood A, Sissons J, Rezaian M, Nolder D, Warthurst D Khan NA. Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates. J Med Mi-crobiol. 2005; 54: 755-759.

Hsu B, Ma P, Liou T, Chen J, Shine F. Identifi-cation of 18S ribosomal DNA genotype of Acanthamoeba from hot spring recreation areas in the central range, Taiwan. J Hydrobiol. 2009; 367: 249-254.


  • There are currently no refbacks.

Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.