Dataset Information

Molecular Identification of Free-Living Amoebae (Naegleria spp., Acanthamoeba spp. and Vermamoeba spp.) Isolated from Un-improved Hot Springs, Guilan Province, Northern Iran.

ABSTRACT: Background: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 ?m. 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.

SUBMITTER: Feiz Haddad MH

PROVIDER: S-EPMC7028243 | biostudies-literature | 2019 Oct-Dec

REPOSITORIES: biostudies-literature

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Publications

Molecular Identification of Free-Living Amoebae (<i>Naegleria</i> spp., <i>Acanthamoeba</i> spp. and <i>Vermamoeba</i> spp.) Isolated from Un-improved Hot Springs, Guilan Province, Northern Iran.

Feiz Haddad Mohammad Hossein MH Khoshnood Saeed S Mahmoudi Mohammad Reza MR Habibpour Habib H Ali Selman A SA Mirzaei Habibollah H Feiz Haddad Rezvan R Ahmadiangali Kambiz K

Iranian journal of parasitology 20191001 4

<h4>Background</h4>This study was conducted to determine the presence and molecular identify of <i>Acanthamoeba</i>, <i>Naegleria</i> and <i>Vermamoeba</i> in unimproved hot springs.<h4>Methods</h4>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 <i>Acanthamoeba</i>, <i>Naegleria</i> and <i>Vermamoeba</i> approximately 500 ml of the water samples were filtered through a cellulose nitrate membrane wi ...[more]

PMID: 32099561

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Project description:BackgroundLegionella spp. employ multiple strategies to adapt to stressful environments including the proliferation in protective biofilms and the ability to form associations with free-living amoeba (FLA). The aim of the current study was to identify Legionella spp., Acanthamoeba spp., Vermamoeba (Hartmannella) vermiformis and Naegleria fowleri that persist in a harvested rainwater and solar pasteurization treatment system.MethodsPasteurized (45 °C, 65 °C, 68 °C, 74 °C, 84 °C and 93 °C) and unpasteurized tank water samples were screened for Legionella spp. and the heterotrophic plate count was enumerated. Additionally, ethidium monoazide quantitative polymerase chain reaction (EMA-qPCR) was utilized for the quantification of viable Legionella spp., Acanthamoeba spp., V. vermiformis and N. fowleri in pasteurized (68 °C, 74 °C, 84 °C and 93 °C) and unpasteurized tank water samples, respectively.ResultsOf the 82 Legionella spp. isolated from unpasteurized tank water samples, Legionella longbeachae (35 %) was the most frequently isolated, followed by Legionella norrlandica (27 %) and Legionella rowbothamii (4 %). Additionally, a positive correlation was recorded between the heterotrophic plate count vs. the number of Legionella spp. detected (ρ = 0.710, P = 0.048) and the heterotrophic plate count vs. the number of Legionella spp. isolated (ρ = 0.779, P = 0.0028) from the tank water samples collected. Solar pasteurization was effective in reducing the gene copies of viable V. vermiformis (3-log) and N. fowleri (5-log) to below the lower limit of detection at temperatures of 68-93 °C and 74-93 °C, respectively. Conversely, while the gene copies of viable Legionella and Acanthamoeba were significantly reduced by 2-logs (P = 0.0024) and 1-log (P = 0.0015) overall, respectively, both organisms were still detected after pasteurization at 93 °C.ConclusionsResults from this study indicate that Acanthamoeba spp. primarily acts as the vector and aids in the survival of Legionella spp. in the solar pasteurized rainwater as both organisms were detected and were viable at high temperatures (68-93 °C).

Dataset Information

Molecular Identification of Free-Living Amoebae (Naegleria spp., Acanthamoeba spp. and Vermamoeba spp.) Isolated from Un-improved Hot Springs, Guilan Province, Northern Iran.

Publications

Molecular Identification of Free-Living Amoebae (<i>Naegleria</i> spp., <i>Acanthamoeba</i> spp. and <i>Vermamoeba</i> spp.) Isolated from Un-improved Hot Springs, Guilan Province, Northern Iran.

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