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Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka

BACKGROUND: The inland freshwater bodies in the North-Western Province of Sri Lanka have ideal environmental conditions for the Naegleria species. Therefore, the presence and prevalence of Naegleria species in the water bodies of North-Western Province were determined by molecular characterization a...

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Autores principales: Gunarathna, Nuwan, Amarasinghe, Anjalie, Wijesundara, Sunil, Iddawela, Devika, Wickramasinghe, Susiji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7951808/
https://www.ncbi.nlm.nih.gov/pubmed/33705485
http://dx.doi.org/10.1371/journal.pone.0248510
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author Gunarathna, Nuwan
Amarasinghe, Anjalie
Wijesundara, Sunil
Iddawela, Devika
Wickramasinghe, Susiji
author_facet Gunarathna, Nuwan
Amarasinghe, Anjalie
Wijesundara, Sunil
Iddawela, Devika
Wickramasinghe, Susiji
author_sort Gunarathna, Nuwan
collection PubMed
description BACKGROUND: The inland freshwater bodies in the North-Western Province of Sri Lanka have ideal environmental conditions for the Naegleria species. Therefore, the presence and prevalence of Naegleria species in the water bodies of North-Western Province were determined by molecular characterization and phylogenetic analysis in this study. METHODS: A total of 104 water bodies were selected from Kurunegala and Puttalam districts in the North-Western Province of Sri Lanka. Mean turbidity, pH, and temperature were recorded in each water body from three selected site. Centrifuged samples were cultured on non-nutrient agar plates with Escherichia coli. Enflagellation test positive isolates were subjected to DNA extraction and polymerase chain reaction using genus and species-specific primers targeting the internal transcribed spacer region (ITS) and Mp2CL5 gene. Phylogenetic analysis was performed using Bayesian and maximum likelihood (ML) methods. RESULTS: The prevalence of Naegleria species and N. fowleri in the study area were 23.07% and 1.92%. The prevalence of Naegleria species and the physicochemical parameters of the water bodies showed no significant correlation. Bayesian analysis of the ITS region revealed the Naegleria Sri Lankan (SL) isolates 1, 3, and 4 in a single clade separated from the 2 and 5. Furthermore, Bayesian analysis identified isolates 2 and 5 in the same clade with Naegleria sp. samples and N. Philippinensis forming a sister clade. However, in the ML tree, all isolates were in the same clade with Naegleria sp. samples and N. Philippinensis. CONCLUSIONS: The present study reports the first isolation of pathogenic N. fowleri from Sri Lanka. Based on Bayesian analysis, SL isolates 2 and 5 form a separate clade from 1, 3, and 4. However, in ML analysis, all isolates are grouped in one clade with Naegleria sp. samples and N. philippinensis. Further investigations are required to confirm these findings.
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spelling pubmed-79518082021-03-22 Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka Gunarathna, Nuwan Amarasinghe, Anjalie Wijesundara, Sunil Iddawela, Devika Wickramasinghe, Susiji PLoS One Research Article BACKGROUND: The inland freshwater bodies in the North-Western Province of Sri Lanka have ideal environmental conditions for the Naegleria species. Therefore, the presence and prevalence of Naegleria species in the water bodies of North-Western Province were determined by molecular characterization and phylogenetic analysis in this study. METHODS: A total of 104 water bodies were selected from Kurunegala and Puttalam districts in the North-Western Province of Sri Lanka. Mean turbidity, pH, and temperature were recorded in each water body from three selected site. Centrifuged samples were cultured on non-nutrient agar plates with Escherichia coli. Enflagellation test positive isolates were subjected to DNA extraction and polymerase chain reaction using genus and species-specific primers targeting the internal transcribed spacer region (ITS) and Mp2CL5 gene. Phylogenetic analysis was performed using Bayesian and maximum likelihood (ML) methods. RESULTS: The prevalence of Naegleria species and N. fowleri in the study area were 23.07% and 1.92%. The prevalence of Naegleria species and the physicochemical parameters of the water bodies showed no significant correlation. Bayesian analysis of the ITS region revealed the Naegleria Sri Lankan (SL) isolates 1, 3, and 4 in a single clade separated from the 2 and 5. Furthermore, Bayesian analysis identified isolates 2 and 5 in the same clade with Naegleria sp. samples and N. Philippinensis forming a sister clade. However, in the ML tree, all isolates were in the same clade with Naegleria sp. samples and N. Philippinensis. CONCLUSIONS: The present study reports the first isolation of pathogenic N. fowleri from Sri Lanka. Based on Bayesian analysis, SL isolates 2 and 5 form a separate clade from 1, 3, and 4. However, in ML analysis, all isolates are grouped in one clade with Naegleria sp. samples and N. philippinensis. Further investigations are required to confirm these findings. Public Library of Science 2021-03-11 /pmc/articles/PMC7951808/ /pubmed/33705485 http://dx.doi.org/10.1371/journal.pone.0248510 Text en © 2021 Gunarathna et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Gunarathna, Nuwan
Amarasinghe, Anjalie
Wijesundara, Sunil
Iddawela, Devika
Wickramasinghe, Susiji
Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka
title Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka
title_full Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka
title_fullStr Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka
title_full_unstemmed Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka
title_short Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka
title_sort isolation, molecular characterization and phylogeny of naegleria species in water bodies of north-western province, sri lanka
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7951808/
https://www.ncbi.nlm.nih.gov/pubmed/33705485
http://dx.doi.org/10.1371/journal.pone.0248510
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