Genotyping and physiological characteristics of Acanthamoeba isolated from beaches in Phuket Province, Thailand

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NARUMON BOONMAN
CHANATE WANNA
JARUWAN CHUTRTONG
SRILERT CHOTPANTARAT
SIRIPHAN BOONSILP

Abstract

Abstract. Boonman N, Wanna C, Chutrtong J, Chotpantarat S, Boonsilp S. 2022. Genotyping and physiological characteristics of Acanthamoeba isolated from beaches in Phuket Province, Thailand. Biodiversitas 23: 4377-4384. Acanthamoeba is widely distributed in water, soil, dust and air. Beaches are popular tourist destinations for water-related activities while they can also be reservoirs for Acanthamoeba to humans. This study aimed to determine the genotyping of Acanthamoeba in the beach environment and characterized their pathogenicity potential. A total of 63 samples were collected from six beaches in Phuket Province, Thailand. Twenty-one samples of each source including wet sand, dry sand and seawater, were collected and cultivated on 2% non-nutrient agar coated with heat-killed Escherichia coli. Acanthamoeba was found in 100% (n=21) wet sand, 52.4% (n=11) dry sand and 14.3% (n=3) seawater. Sequence analysis of the GTSA.B1 region in 18S rRNA revealed that 83.9% of sequenced isolates belonged to the T5 genotype and 16.1% belonged to the T4 genotype. The T5 genotype was predominant, differing from other reports of Acanthamoeba prevalence in natural sources. In vitro thermal and salt-tolerance showed that all isolates had growth rates similar to clinical Acanthamoeba isolates. This research was undertaken to increase awareness of human eye and brain infections caused by Acanthamoeba on beaches. Further studies on these Acanthamoeba isolates are still required such as co-culture assay between the trophozoites and human cell lines to confirm their pathogenicity as well as their susceptibility to the chemotherapeutic agents.

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Bonilla TD, Nowosielski K, Cuvelier M, Hartz A, Green M, Esiobu N, McCorquodale DS, Fleisher JM, Rogerson A. 2007. Prevalence and distribution of fecal indicator organisms in South Florida beach sand and preliminary assessment of health effects associated with beach sand exposure. Mar Pollut Bull. 54(9): 1472-1482. DOI: 10.1016/j.marpolbul.2007.04.016

Boonman N, Prachya S, Boonmee A, Kittakoop P, Wiyakrutta S, Sriubolmas N, Warit S, Chusattayanond AD. 2012. In vitro acanthamoebicidal activity of fusaric acid and dehydrofusaric acid from an endophytic fungus Fusarium sp. Tlau3. Planta Med. 78(14): 1562-1567. DOI: 10.1055/s-0032-1315146

Booton GC, Rogerson A, Bonilla TD, Seal DV, Kelly DJ, Beattie TK, Tomlinson A, Lares-villa F, Fuerst PA, Byers TJ. 2004. Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis. J Eukaryot Microbiol. 51(2): 192-200. DOI: 10.1111/j.1550-7408.2004.tb00545.x.

de Lacerda AG, Lira M. 2021. Acanthamoeba keratitis: a review of biology, pathophysiology and epidemiology. Ophthalmic Physiol Opt. 41(1):116-135. DOI: 10.1111/opo.12752

Fanselow N, Sirajuddin N, Yin XT, Huang AJ, Stuart PM. 2021. Acanthamoeba keratitis, pathology, diagnosis and treatment. Pathogens. 10(3): 323. DOI: 10.3390/pathogens10030323

Fuerst PA, Booton GC, Crary M. 2015. Phylogenetic analysis and the evolution of the 18S rRNA gene typing system of Acanthamoeba. J Eukaryot Microbiol. 62(1): 69-84. DOI: 10.1111/jeu.12186

Halliday E, Gast RJ. 2011. Bacteria in beach sands: an emerging challenge in protecting coastal water quality and bather health. Environ Sci Technol. 45(2): 370-379. DOI: 10.1021/es102747s

Kalra SK, Sharma P, Shyam K, Tejan N, Ghoshal U. 2020. Acanthamoeba and its pathogenic role in granulomatous amebic encephalitis. Exp. Parasitol. 208: 107788. DOI: 10.1016/j.exppara.2019.107788

Khan NA, Jarroll EL, Paget TA. 2002. Molecular and physiological differentiation between pathogenic and nonpathogenic Acanthamoeba. Curr Microbiol. 45(3): 197-202. DOI: 10.1007/s00284-001-0108-3

Maciver SK, Asif M, Simmen MW, Lorenzo-Morales J. 2013. A systematic analysis of Acanthamoeba genotype frequency correlated with source and pathogenicity: T4 is confirmed as a pathogen-rich genotype. Eur J Protistol. 49(2): 217-221. DOI: 10.1016/j.ejop.2012.11.004

Mahmoudi MR, Karanis P. 2020. Genotyping of Acanthamoeba spp. isolated from the Caspian Sea in Iran. Environ. Sci. Proc. 2(1): 9. DOI: 10.3390/environsciproc2020002009

Mohd Hussain RH, Abdul Ghani MK, Khan NA, Siddiqui R, Anuar TS. 2022. Acanthamoeba species isolated from marine water in Malaysia exhibit distinct genotypes and variable physiological properties. J Water Health. 20(1): 54-67. DOI: 10.2166/wh.2021.128

Paknejad N, Hajialilo E, Saraei M, Javadi A. 2020. Isolation and identification of Acanthamoeba genotypes and Naegleria spp. from the water samples of public swimming pools in Qazvin, Iran. J Water Health. 18(2): 244-251. DOI: 10.2166/wh.2019.074

Possamai CO, Loss AC, Costa AO, Falqueto A, Furst C. 2018. Acanthamoeba of three morphological groups and distinct genotypes exhibit variable and weakly inter-related physiological properties. Parasitol Res. 117(5): 1389-1400. DOI: 10.1007/s00436-018-5824-8

Putaporntip C, Kuamsab N, Nuprasert W, Rojrung R, Pattanawong U, Tia T, Yanmanee S, Jongwutiwes S. 2021. Analysis of Acanthamoeba genotypes from public freshwater sources in Thailand reveals a new genotype, T23 Acanthamoeba bangkokensis sp. nov. Sci Rep. 11(1): 17290. DOI: 10.1038/s41598-021-96690-0

Qvarnstrom Y, Nerad TA, Visvesvara GS. 2013. Characterization of a new pathogenic Acanthamoeba species, A. byersi n. sp., isolated from a human with fatal amoebic encephalitis. J Eukaryot Microbiol. 60(6): 626-633. DOI: 10.1111/jeu.12069

Schroeder JM, Booton GC, Hay J, Niszl IA, Seal DV, Markus MB, Fuerst PA, Byers TJ. 2001. Use of subgenic 18S ribosomal DNA PCR and sequencing for genus and genotype identification of Acanthamoebae from humans with keratitis and from sewage sludge. J Clin Microbiol. 39(5): 1903-1911. DOI: 10.1128/JCM.39.5.1903-1911.2001

Shyrobokov V, Poniatovskyi V, Chobotar A, Salamatin R. 2020. Morphological, physiological and genetic characteristics of protozoa of genus Acanthamoeba, isolated from different deposit of bentonite in Ukraine. Ann. Parasitol. 66(1): 69–75. DOI: 10.17420/ap6601.239

Sudjaroen Y, Thongkao K, Suwannahong K. 2017. Characteristics of mineral water from nature hot springs in Ranong Province, Thailand. Ann Trop Med Public Health. 10(2): 366-370. DOI: 10.4103/1755-6783.196588

Szentmáry N, Daas L, Shi L, Laurik KL, Lepper S, Milioti G, Seitz B. 2019. Acanthamoeba keratitis–Clinical signs, differential diagnosis and treatment. J Curr Ophthalmo. 31(1): 16-23. DOI: 10.1016/j.joco.2018.09.008

Trabelsi H, Dendana F, Sellami A, Sellami H, Cheikhrouhou F, Neji S, Makni A, Ayadi A. 2012. Pathogenic free-living amoebae: epidemiology and clinical review. Pathologie Biologie. 60(6): 399-405. DOI: 10.1016/j.patbio.2012.03.002

Walochnik J, Scheikl U, Haller?Schober EM. 2015. Twenty years of Acanthamoeba diagnostics in Austria. J Eukaryot Microbiol. 62(1): 3-11. DOI: 10.1111/jeu.12149

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