Articles

Pathogenic Assays of Acanthamoeba Belonging to the T4 Genotype

Abstract

 

Background: Acanthamoeba genus is introduced as opportunistic and cosmopoli-tan parasite. Monkey and wistar rat are appropriate models for experimental study on Acanthamoeba infection. In this study Acanthamoeba spp. were isolated from hot spring (HS), windows dust (WD) and a corneal sample of keratitis patient (KP) and their pathogenicity surveyed by in vitro and in vivo tests.

Methods: Isolates of Acanthamoeba were cultivated axenically for 12 months in PYG medium. Overall, 30 wistar rats, in 6 equal groups were used for developing experimental Acanthamoeba keratitis (AK) and Granulomatous Amoebic Encephalitis (GAE). The Keratitis and Granulomatous Encephalitis experiments were per-formed by intrastromal and intranasal inoculation of Acanthamoeba cysts, respec-tively. Pathogenicity of the three isolates was also evaluated by in vitro test using osmotolerance and temperature tolerance assays. Identification of genotypes were performed by PCR technique and sequencing.

Result: None of the isolates could perform AK and GAE in wistar rats, although all isolates were described as T4 genotype. Isolates obtained from KP and WD could grow only in 30 °C, but not in 37 °C and 40 °C. On the other hand, HS iso-late grew in 30 °C and 37 °C but not in 40 °C. Moreover, all of isolate grew in 0.5 M mannitol but not in 1 M and 1.5 M.

Conclusion: T4 isolates with a long-term axenic culture and different factors re-lated to host and parasite may play role in pathogenicity of these free-living amoebae.

Garajova M, Mrva M. [Amoebae of the genus Acanthamoeba--causative agents of human infec-tions]. Epidemiol Mikrobiol Imunol. 2011;-60(3):121-30.

Siddiqui R, Khan NA. Biology and pathoge-nesis of Acanthamoeba. Parasit Vectors. 2012;5:6.

Khan NA. Pathogenesis of Acanthamoeba infections. Microb Pathog. 2003;34(6):277-85.

Niyyati M, Lorenzo-Morales J, Rahimi F, Motevalli-Haghi A, Martin-Navarro CM, Far-nia S, et al. Isolation and genotyping of potentially pathogenic Acanthamoeba strains from dust sources in Iran. Trans R Soc Trop Med Hyg. 2009;103(4):425-7.

Clarke DW, Niederkorn JY. The pathoph-ysiology of Acanthamoeba keratitis. Trends Parasitol. 2006;22(4):175-80.

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

Khan NA. Acanthamoeba invasion of the central nervous system. Int J Parasitol. 2007;37(2): 131-8 .

Martinez AJ. Infection of the central nervous system due to Acanthamoeba. Rev Infect Dis. 1991;13 Suppl 5:S399-402.

Kong HH. Molecular phylogeny of Acanth-amoeba. Korean J Parasitol. 2009;47 Suppl:S21-8.

Nuprasert W, Putaporntip C, Pariyakanok L, Jongwutiwes S. Identification of a novel t17 genotype of Acanthamoeba from environmental isolates and T10 genotype causing keratitis in Thailand. J Clin Microbiol. 2010;48(12):4636.

Corsaro D, Venditti D. Phylogenetic evidence for a new genotype of Acanthamoeba (Amoebozoa, Acanthamoebida). Parasitol Res. 2010;107(1):233.

Badirzadeh A, Niyyati M, Babaei Z, Amini H, Badirzadeh H, Rezaeian M. Isolation of free-living amoebae from sarein hot springs in ardebil province, Iran. Iranian J Parasitol. 2011;6(2):1-8.

Larkin DF, Easty DL. Experimental Acanthamoeba keratitis: I. Preliminary findings. Br J Ophthalmol. 1990;74(9):551-5.

Polat ZA, Ozcelik S, Vural A, Yildiz E, Cetin A. Clinical and histologic evaluations of experi-mental Acanthamoeba keratitis. Parasitol Res. 2007;101(6):1621-5.

Ren M, Wu X. Evaluation of three different methods to establish animal models of Acan-thamoeba keratitis. Yonsei Med J. 2010;-51(1):121-7.

Walochnik J, Obwaller A, Aspock H. Correlations between morphological, molecular biological, and physiological characteristics in clinical and nonclinical isolates of Acanthamoeba spp. Appl Environ Microbiol. 2000;66-(10):4408.

Khan NA, Jarroll EL, Paget TA. Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays. Curr Microbiol. 2001;43(3):204-8.

De Jonckheere JF. Growth characteristics, cytopathic effect in cell culture, and virulence in mice of 36 type strains belonging to 19 differ-ent Acanthamoeba spp. Appl Environ Microbiol. 1980;39(4):681-5.

Lasjerdi Z, Niyyati M, Haghighi A, Shahabi S, Biderouni FT, Taghipour N, et al. Potentially pathogenic free-living amoebae isolated from hospital wards with immunodeficient patients in Tehran, Iran. Parasitol Res. 2011;109(3):575.

Nuprasert W, Putaporntip C, Pariyakanok L, Jongwutiwes S. Identification of a novel T17 genotype of Acanthamoeba from environ-mental isolates and T10 genotype causing keratitis in Thailand. J Clin Microbiol. 2010; 48 (12): 4636–40.

Booton GC, Visvesvara GS, Byers TJ, Kelly DJ, Fuerst PA. Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. J Clin Microbiol. 2005;43(4):1689-93.

Niyyati M, Lorenzo-Morales J, Rezaie S, Rahimi F, Mohebali M, Maghsood AH, et al. Genotyping of Acanthamoeba isolates from clinical and environmental specimens in Iran. Exp Parasitol. 2009;121(3):242-5.

Maghsood AH, Sissons J, Rezaian M, Nolder D, Warhurst D, Khan NA. Acanthamoeba genotype T4 from the UK and Iran and isolation of the T2 genotype from clinical isolates. J Med Microbiol. 2005;54(Pt 8):755-9. 24. Rahdar M, Niyyati M, Salehi M, Feghhi M, Mak-vandi M, Pourmehdi M, Farnia S. Isolation and genotyping of Acanthamoeb-a strains from environ-mental sources in ahvaz city, khuzestan province, southern Iran. Iranian J Parasitol. 2012;7(4):22-6

Stothard DR, Schroeder-Diedrich JM, Awwad MH, Gast RJ, Ledee DR, Rodriguez-Zaragoza S, et al. The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J Eukaryot Microbiol. 1998;45(1):45-54. 26. Maciver SK, Asif M, Simmen MW, Lorenzo-Morales J. A systematic analysis of Acantha-moeba genotype frequency correlated with source and pathogenicity: T4 is confirmed as a pathogen-rich genotype. Eur J Protistol.- 2013;49(2):217.

Khan NA, Jarroll EL, Paget TA. Molecular and Physiological Differentiation Between Pathogenic and Nonpathogenic Acanthamoeba. Curr Microbiol. 2002;45(3):197-202.

Khan NA. Pathogenicity, morphology, and differ-entiation of Acanthamoeba. Curr Micro-biol. 2001;43(6):391-5.

Walochnik J, Haller-Schober E, Kolli H, Picher O, Obwaller A, Aspock H. Discrimination between clinically relevant and nonrelevant Acanthamoeba strains isolated from contact lens- wearing keratitis patients in Austria. J Clin Microbiol. 2000;38(11):3932-6.

Stevens AR, O'Dell WD. In vitro growth and virulence of Acanthamoeba. J Parasitol. 1974;-60(5):884-5.

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IssueVol 8 No 4 (2013) QRcode
SectionArticles
Keywords
Acanthamoeba T4 Experimental Model Osmotolerance Pathogenicity Rat

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How to Cite
1.
Mirjalali H, Niyyati M, Abedkhojasteh H, Babaei Z, Sharifdini M, Rezaeian M. Pathogenic Assays of Acanthamoeba Belonging to the T4 Genotype. Iran J Parasitol. 1;8(4):530-535.