Axenic Cultivation and Pathogenic Assays of Acanthamoeba Strains Using Physical Parameters
Abstract
Background: The main goal of the present study was to set up an axenic cultivation of Acanthamoeba and assess the pathogenic ability of T4 genotypes from different clinical and environmental strains of Acanthamoeba using two physical assays.
Methods: Sixteen Acanthamoeba isolates including 10 environmental and 6 clinical strains were cul-tured axenically. Axenic cultivation was performed using Proteosepepton, yeast extract and glucose medium and TY-I-S33culture. Pathogenic survey was done using osmotolerance and thermotoler-ance assay. Briefly, differentosmolarity (0.5 M and 1 M) of non-nutrient agar plates were performed. One hundred fiftyμl of axenic culture were collected and were inoculated in 1% agar medium. For thermotolerance assay 150 μl of amoebas from axenic culture were divided into fresh culture me-diums. Cultures were incubated at 37oC and 42 oC. All plates were monitored for 24 h, 48 h and 72 h.
Results: Overall, 16 strains of Acanthamoeba isolates previously genotyped as T4 were cultivated axenically after several months. Thermotolerance and osmotolerance assay revealed that all of clinical strains, soil and animal feces strains were highly pathogenic isolates. Two dust and water strains did not grow at high temperature (42 oC) and osmolarity (1.5 M) and thus they were classified as weak pathogens.
Conclusion: Most of T4 genotypes are highly pathogenic organisms. This is an important finding since Acanthamoeba belonging to T4 type is the predominate genotype in environmental and clinical samples. The presence of highly pathogenic Acanthamoeba may pose a risk within susceptible people
Khan NA. Acanthamoeba: biology and increasing importance in human health. FEMS Microbiol Rev. 2006; 30:564-595.
Marciano-Cabral F, Cabral G. Acanthamoeba spp. as agents of disease. Clin Microbiol Rev. 2003; 16.
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: 4636.
Khan NA. Acanthamoeba, biology and patho-genesis, 1st ed. Caister Academic Press. 2009.
Khan NA, Jarroll EL, Paget TA. Acanthamoeba can be differentiated by the Polymerase Chain Reaction and simple plating assays. Curr Microbiol. 2001; 43: 204-208.
Dejonkheere JF. Growth characteristics, cyto-patic effect in cell culture and virulence in mice belonging to 19 different Acanthamoeba SPP. Appl Environ Microbiol. 1980; 39: 681-685.
Niyyati M, Lorenzo-Morales J, Rezaie s, Rahimi F, Mohebali M, Maghsood AH, Motevalli-Haghi A, Martín-NavarroCM, Farnia S,Valladares B, Rezaeian M. Genotyping of Acanthamoeba isolates from clinical and environmental specimens in Iran. Exp Parasitol. 2009; 121(3): 242-245.
Niyyati M, Lorenzo-Morales J, Rahimi F, Motevalli-Haghi A, Martín-Navarro CM, Far-niaSh, Valladares B, Rezaeian M. Isolation and genotyping of potentially pathogenic Acan-thamoeba strains from dust sources in Iran.Trans R Soc Trop Med Hyg.2009; 103.
Rahdar M, Niyyati M, Salehi M, Feghhi M, Mak-vandi M, Poormehdi M, Farnia Sh. Isolation and genotyping of Acanthamoeba strains, Khuzestan province. Iranian J Parasitol. In press.
Clark CG, Diamond LS. Methods for Cultiva-tion of Luminal Parasitic Protists of Clinical Im-portance.Clin Microbiol Rev. 2002; 15(3).
Khan NA, Jarroll EL, Paget TA. Molecular and physiological differentiation between patho-genic and non-pathogenic Acanthamoeba. Curr Microbiol. 2002; 45: 197-202.
| Files | ||
| Issue | Vol 8 No 2 (2013) | |
| Section | Articles | |
| Keywords | ||
| Acanthamoeba Axenic cultivation Pathogenicity | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
