Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka.
<h4>Background</h4>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...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2021-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0248510 |
| _version_ | 1819035073573289984 |
|---|---|
| author | Nuwan Gunarathna Anjalie Amarasinghe Sunil Wijesundara Devika Iddawela Susiji Wickramasinghe |
| author_facet | Nuwan Gunarathna Anjalie Amarasinghe Sunil Wijesundara Devika Iddawela Susiji Wickramasinghe |
| author_sort | Nuwan Gunarathna |
| collection | DOAJ |
| description | <h4>Background</h4>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.<h4>Methods</h4>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.<h4>Results</h4>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.<h4>Conclusions</h4>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. |
| first_indexed | 2024-12-21T07:43:50Z |
| format | Article |
| id | doaj.art-62d908f5143147a7b4a5fff24d123e76 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T07:43:50Z |
| publishDate | 2021-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-62d908f5143147a7b4a5fff24d123e762022-12-21T19:11:15ZengPublic Library of Science (PLoS)PLoS ONE1932-62032021-01-01163e024851010.1371/journal.pone.0248510Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka.Nuwan GunarathnaAnjalie AmarasingheSunil WijesundaraDevika IddawelaSusiji Wickramasinghe<h4>Background</h4>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.<h4>Methods</h4>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.<h4>Results</h4>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.<h4>Conclusions</h4>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.https://doi.org/10.1371/journal.pone.0248510 |
| spellingShingle | Nuwan Gunarathna Anjalie Amarasinghe Sunil Wijesundara Devika Iddawela Susiji Wickramasinghe Isolation, molecular characterization and phylogeny of Naegleria species in water bodies of North-Western Province, Sri Lanka. PLoS ONE |
| 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 |
| url | https://doi.org/10.1371/journal.pone.0248510 |
| work_keys_str_mv | AT nuwangunarathna isolationmolecularcharacterizationandphylogenyofnaegleriaspeciesinwaterbodiesofnorthwesternprovincesrilanka AT anjalieamarasinghe isolationmolecularcharacterizationandphylogenyofnaegleriaspeciesinwaterbodiesofnorthwesternprovincesrilanka AT sunilwijesundara isolationmolecularcharacterizationandphylogenyofnaegleriaspeciesinwaterbodiesofnorthwesternprovincesrilanka AT devikaiddawela isolationmolecularcharacterizationandphylogenyofnaegleriaspeciesinwaterbodiesofnorthwesternprovincesrilanka AT susijiwickramasinghe isolationmolecularcharacterizationandphylogenyofnaegleriaspeciesinwaterbodiesofnorthwesternprovincesrilanka |