Molecular Identification of Free-Living Amoebae (Naegleria spp., Acanthamoeba spp. and Vermamoeba spp.) Isolated from Unimproved Hot Springs, Guilan Province, Northern Iran
AbstractBackground: This study was conducted to determine the presence and molecular identify of Acanthamoeba, Naegleria and Vermamoeba in unimproved hot springs. Methods: From Jul to Aug 2017, 54 water samples were collected from hot springs in different parts of the Guilan Province, North Iran. For the isolation of Acanthamoeba, Naegleria and Vermamoeba approximately 500 ml of the water samples were filtered through a cellulose nitrate membrane with a pore size of 0.45 μm. The filter was transferred onto non-nutrient agar plates seeded with Gram-negative bacteria (Escherichia coli) as a food source. The morphological key of page was used to identify free‐living amoebae (FLA) using an inverted microscope, PCR amplification targeting specific genes for each genus and sequencing determined frequent species and genotypes base on NCBI database. Results: Fifteen of the 54 samples were positive by culture and/or PCR for Acanthamoeba and other FLA from unimproved hot springs. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba castellanii (12 case isolates belonged to T4 genotype), 4 cases of V. vermiformis, and 3 cases of N. australiensis, 2 cases of N. pagei and 1 cases of N. gruberi. Conclusion: Although FLA-mediated illnesses are not as high as in environmental distribution, but because of a poor prognosis, more investigations about FLA distribution in hot springs is critical. Hot spring may enhance exposure of the amoebae in individuals. Hence, more attention to unimproved hot springs is needed to prevent free-living amoebae mediated diseases.
2. Javanmard E, Niyyati M, Lorenzo-Morales J et al. Molecular identification of waterborne free living amoebae (Acanthamoeba, Naegleria and Vermamoeba) isolated from municipal drinking water and environmental sources, Semnan province, north half of Iran. Exp Parasitol. 2017;183:240-244.
3. Niyyati M, Lorenzo-Morales J et al. First report of a mixed infection due to Acanthamoeba genotype T3 and Vahlkampfia in a cosmetic soft contact lens wearer in Iran. Exp Parasitol. 2010;126(1):89-90.
4. Lorenzo-Morales J, Martínez-Carretero E, Batista N et al. Early diagnosis of amoebic keratitis due to a mixed infection with Acanthamoeba and Hartmannella. Parasitol Res. 2007;102(1):167-9.
5. Abedkhojasteh H, Niyyati M, Rahimi F et al. First report of Hartmannella keratitis in a cosmetic soft contact lens wearer in Iran. Iran J Parasitol. 2013;8(3):481-5.
6. Khan NA. Acanthamoeba: biology and pathogenesis: Horizon Scientific Press; 2009.
7. Visvesvara GS. Chapter 10 - Infections with free-living amebae. In: Garcia HH, Tanowitz HB, Del Brutto OH, editors. Handb Clin Neurol. 114: Elsevier; 2013. p. 153-68.
8. De Jonckheere JF. What do we know by now about the genus Naegleria? Exp Parasitol. 2014;145 Suppl:S2-9.
9. Zbikowska E, Walczak M, Krawiec A. Distribution of Legionella pneumophila bacteria and Naegleria and Hartmannella amoebae in thermal saline baths used in balneotherapy. Parasitol Res. 2013;112(1):77-83.
10. Scheid P. Relevance of free-living amoebae as hosts for phylogenetically diverse microorganisms. Parasitol Res. 2014;113(7):2407-14.
11. Niyyati M, Lasgerdi Z, Lorenzo-Morales J. Detection and molecular characterization of potentially pathogenic free-living amoebae from water sources in Kish Island, Southern Iran. Microbiol Insights. 2015;8(Suppl 1):1-6.
12. Badirzadeh A, Niyyati M, Babaei Z et al. Isolation of free-living amoebae from sarein hot springs in ardebil province, iran. Iran J Parasitol. 2011;6(2):1-8.
13. R Solgi, M Niyyati, A Haghighi et al. Occurrence of thermotolerant Hartmannella vermiformis and Naegleria spp. in hot springs of Ardebil Province, Northwest Iran. Iran J Parasitol. 2012; 7(2): 47–52.
14. Latifi AR, Niyyati M, Lorenzo-Morales J et al. Occurrence of Naegleria species in therapeutic geothermal water sources, Northern Iran. Acta Parasitol. 2017;62(1):104-109.
15. Mahmoudi, M.R., Kazemi, B., Haghighi, A., Karanis, P. Detection of Acanthamoeba and Toxoplasma in river water samples by molecular methods in Iran. Iran J Parasitol. 10(2), pp. 250-257.
16. Solgi R, Niyyati M, Haghighi A et al. Thermotolerant Acanthamoeba spp. isolated from therapeutic hot springs in Northwestern Iran. J Water Health. 2012;10(4):650-6.
17. Feiz Haddad MH, Ghasemi E, Maraghi S, Tavala M. Identification of Leishmania Species Isolated from Human Cutaneous Leishmaniasis in Mehran, Western Iran Using Nested PCR. Iran J Parasitol. 2016;11(1):65-72.
18. Page FC. A new key to freshwater and soil Gymnamoebae: with instructions for culture: Freshwater Biological Association; 1988.
19. Hossein Hooshyar, Bahram Hosseinbigi, Mehrzad Saraei et al. Genotyping of Acanthamoeba isolated from surface and stagnant waters of Qazvin, Central Iran. Iran Red Crescent Med J. 2013;15(6): 536–538.
20. Lasjerdi Z, Niyyati M, Haghighi A et al. Potentially pathogenic free-living amoebae isolated from hospital wards with immunodeficient patients in Tehran, Iran. Parasitol Res. 2011;109(3):575-80.
21. Nazar M, Haghighi A, Niyyati M et al. Genotyping of Acanthamoeba isolated from water in recreational areas of Tehran, Iran. J Water Health. 2011;9(3):603-8.
22. Niyyati M, Lorenzo-Morales J, Rezaie S et al. Genotyping of Acanthamoeba isolates from clinical and environmental specimens in Iran. Exp Parasitol. 2009;121(3):242-5.
23. Feiz-Haddad MH, Shokri A, Habibpour H, Nejadi SM. A review of Acanthamoeba keratitis in the Middle East and Iran. Journal of Acute Disease. 2019;8(4):133-41.
24. Edagawa A, Kimura A, Kawabuchi-Kurata T et al. Isolation and genotyping of potentially pathogenic Acanthamoeba and Naegleria species from tap-water sources in Osaka, Japan. Parasitol Res. 2009;105(4):1109-17.
25. Movahedi Z, Shokrollahi MR, Aghaali M et al. Primary amoebic meningoencephalitis in an Iranian infant. Case Rep Med. 2012;2012.
26. John DT, De Jonckheere JF. Isolation of Naegleria australiensis from an Oklahoma lake. J Protozool. 1985;32(4):571-5.
27. Hajialilo E, Niyyati M, Solaymani M, Rezaeian M. Pathogenic free-living amoebae isolated from contact lenses of keratitis patients. Iran J Parasitol. 2015;10(4):541-6.
28. Mahmoudi MR, Rahmati B, Seyedpour SH, Karanis P. Occurrence and molecular characterization of free-living amoeba species (Acanthamoeba, Hartmannella, and Saccamoeba limax) in various surface water resources of Iran. Parasitol Res. 2015;114(12):4669-74.
29. Feiz-Haddad M-H, Kassiri H, Kasiri N et al. Prevalence and epidemiologic profile of acute cutaneous leishmaniasis in an endemic focus, Southwestern Iran. J Acute Dis. 2015;4(4):292-7.
Copyright (c) 2019 Iranian Journal of Parasitology
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.