Isolation and identification of free-living amoeba from the hot springs and beaches of the Caspian Sea (original) (raw)
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Isolation of free-living amoebae from sarein hot springs in ardebil province, iran
2011
Background: Free-living amoebae (FLA) are a group of ubiquitous protozoan, which are distributed in the natural and artificial environment sources. The main aim of the current study was to identify the presence of FLA in the recreational hot springs of Sarein in Ardebil Province of Iran. Methods: Seven recreational hot springs were selected in Sarein City and 28 water samples (four from each hot spring) were collected using 500 ml sterile plastic bottles during three month. Filtration of water samples was performed, and culture was done in non-nutrient agar medium enriched with Escherichia coli. Identification of the FLA was based on morphological criteria of cysts and trophozoites. Genotype identification of Acanthamoeba positive samples were also performed using sequencing based method. Results: Overall, 12 out of 28 (42.9%) samples were positive for FLA which Acanthamoeba and Vahlkampfiid amoebae were found in one (3.6%) and 11 (39.3%) samples, respectively. Sequence analysis of the single isolate of Acanthamoeba revealed potentially pathogenic T 4 genotype corresponding to A. castellanii. Conclusion: Contamination of hot springs to FLA, such as Acanthamoeba T 4 genotype (A. castellanii) and Vahlkampfiid amoebae, could present a sanitary risk for high risk people, and health authorities must be aware of FLA presence.
Thermotolerant Acanthamoeba spp. isolated from therapeutic hot springs in northwestern Iran
Journal of Water and Health, 2012
This study was conducted to address the distribution of Acanthamoeba genotypes in therapeutic hot springs in Iran. Sixty water and sediment samples were collected from bicarbonate, sulphur, and sodium chloride thermal springs in the northwest. All hot springs examined are used mainly for health purposes in Iran. Acanthamoeba were identified by both morphology and PCR (polymerase chain reaction). Genotype identification was based on the sequencing of a highly variable and informative region of Diagnostic Fragment 3 (stem 29-1 of 18S rRNA gene) within Acanthamoebaspecific amplimer (ASA.S1). Twenty percent of hot springs were contaminated with thermotolerant Acanthamoeba belonging to the potentially pathogenic T4 and T3 genotypes. A high number (91.7%) of strains showed growth at 37 W C, and eight isolates showed growth at 42 W C. A single isolate (HSNW2) was detected in waters at 70 W C. The presence of thermotolerant Acanthamoeba highlights a risk factor for susceptible individuals, as Acanthamoeba-related keratitis continues to rise in Iran.
Detection of free living amoebae, Acanthamoeba and Naegleria, in swimming pools, Malaysia
Tropical Biomedicine
This study reports the detection of Acanthamoeba and Naegleria species in 14 swimming pools around Petaling Jaya and Kuala Lumpur, Malaysia. Sampling was carried out at 4 sites (the platforms (P), wall (W), 1 meter from the wall (1) and middle (2)) of each swimming pool. These free living amoebae (FLA) were detected under light and inverted microscopes after being cultured on the surface of non-nutrient agar lawned with Escherichia coli. Acanthamoeba species were detected in higher number of culture plates from all sampling sites of all the swimming pools. While Naegleria, were detected in fewer culture plates at 3 sampling sites (absent at site P) of 8 swimming pools. This suggested that the thick double-walled cysts of Acanthamoeba were more resistant, thus remaining viable in the dry-hot areas of the platforms and in chlorinated water of the swimming pools whereas Naegleria cysts, that are fragile and susceptible to desiccation, preferred watery or moist areas for growth and proliferation. The prevalence of both FLA was highest at site W (76.2%), followed by site 1 (64.7%), lowest at site 2 (19.4%), and could be detected at all 3 sampling levels (top, middle and bottom) of these 3 sites. The surface of site W might act as a bio-film that accumulated all kinds of microbes providing sufficient requirement for the FLA to develop and undergo many rounds of life cycles as well as moving from top to bottom in order to graze food. Other factors such as human activities, the circulating system which was fixed at all swimming pools, blowing wind which might carry the cysts from surroundings and the swimming flagellate stage of Naegleria, could also contribute to the distribution of the FLA at these sampling sites. Both FLA showed highest growth (80.4%) at room temperature (25-28 degrees C) and lesser (70.0%) at 37 degrees C which might be due to the overgrowth of other microbes (E. coli, fungi, algae, etc). While at 44 degrees C, only Acanthamoeba species could survive thus showing that our swimming pools are free from potentially pathogenic Naegleria species. However, further study is needed in order to confirm the virulence levels of these amoebae isolates.
Current World Environment, 2015
Free living amoebas are widespread protozoans that can be found in different environmental resources such as water bodies, soil and clays. Free living amoebae particularly Acanthamoeba could lead to diseases with poor prognosis including ameobic keratitis (AK) and granulomatous amebic encephalitis (GAE) especially in contact lens wears and immunocompromised individuals, respectively. In the present study 50 samples were collected from water resources (hot-springs, cold springs, rivers, wells, refined drinking water and subterranean canals) of Kaleybar and Khodaafarin, East Azarbayjan Province, Iran. The samples were transferred to laboratory and cultured on 1.5% nonnutrient agar medium containing Escherichia coli bacteria at room temperature and finally examined by microscopic methods. The results showed that out of 50 water samples, 40% were positive samples for free-living amoebae (FLA) including 60% Acanthamoebae genus, %35 Haetmannellidae, %20 Valkamphiids, %20 Vannella, %10 Thechamoeba (45% of plates had mix contamination). The present research was the first study to address the presence of potentially pathogenic free living amoebae in Kaleybar and Khodaafarin, East-Azerbaijan Province.
Journal of Water and Health
Free-living amoebae (FLA) include many genera which cause serious diseases such as sight-threatening keratitis, cutaneous ulcers and fatal encephalitis. This study was conducted due to the lack of research regarding genotypes Acanthamoeba, Naegleria and Vermamoeba in mineral springs of Guilan Province in northern Iran. Twenty-five water samples were collected from mineral springs in Guilan Province. After filtration through nitrocellulose membrane, samples were cultured on non-nutrient agar plates. The morphological key of Page was used to identify free-living amoebae (FLA) using an inverted microscope. Positive cultures were analyzed by polymerase chain reaction (PCR) and genotypes based on the NCBI database. Eleven (44%) samples were positive for Acanthamoeba, Naegleria and Vermamoeba. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba castellanii (three case isolates belonged to T4 genotype), three cases of Vermamoeba vermiformis, and two cases ...
Pathogenic amoebae in natural thermal waters of three resorts of Hidalgo, Mexico
Environmental Research, 1989
In a search for free-living amoebae, seven water samples from three thermal water bathing resorts in Tecozautla, Hidalgo, were analyzed during December 1984. The samples were concentrated by filtration and centrifugation, and inoculated later on monoxenic and axenic media. The identification of the isolates was performed by morphology and isoelectric focusing of isoenzymes and total proteins. Thirty-three strains of free-living amoebae belonging to the genera Naegleria, Acanthamoeba, and Willaertia were isolated. Twenty of these strains belonged to the Naegleria genus, 16 of them were classified as Naegleria spp., and 2 were classified as Naegleria lovaniensis. Noteworthy was the finding of two pathogenic strains of the species Naegleria australiensis. N. australiensis and N. lovaniensis may be considered good indicator organisms, since they live in the same environmental conditions as N. fowleri, the agent of primary amoebic encephalitis (PAM). On the other hand, amoebae other than Naegleria were isolated and identified as Acathamoeba castellanii (two strains), and Acanthamoeba lugdunensis (one strain), which proved to be pathogenic when tested in mice. Nine more pathogenic strains of the genus Acanthamoeba spp. were isolated together with one strain of Willaertia magna, a thermophilic nonpathogenic amoeba. The chlorination and periodical surveillance of water resorts like the one studied is recommended, in order to prevent the appearance of more cases of PAM or other human diseases associated with pathogenic Acanthamoeba spp.
Iranian journal of parasitology
Swimming in contaminated water was reported to be associated with Acanthamoeba and N. fowleri human infections. The present study was carried out with the aim of isolation and identification of the different species of Acanthamoeba and Naegleria from two swimming pools in Alexandria University. Samples were collected from the swimming pools of Alexandria University Stadium and Faculty of Agriculture-Alexandria University during the period from May 2012 to April 2013. Free-living amoebae were prevalent in the collected samples. Molecular characterization confirmed the identity of ten Acanthamoeba isolates and seven Naegleria isolates. Acanthamoeba T3, T4, T5, T11 and T15 genotypes were identified. Acanthamoeba T4 was the most prevalent genotype. The relatively high prevalence of Acanthamoeba, especially genotype T4, indicates the presence of a health hazard to swimmers particularly those wearing contact lenses. Naegleria fowleri was not found during the present study.
Genotyping of Acanthamoeba spp. Isolated from the Caspian Sea in Iran
Environmental Sciences Proceedings, 2020
Acanthamoeba spp. are widely distributed in the environment and have been reported to be causative agents of lethal encephalitis and keratitis. In this study, thirty water samples from the Caspian Sea were collected during 2018. Water samples were filtrated and the filtrate used for culture. The positive samples were subjected to Polymerase Chain Reaction (PCR) and nucleotide sequencing. Free-living amoebae were identified in 50% (15/30) of the seawater samples. DNA sequencing revealed the presence of T2 and T4 genotypes. The results of the present study confirmed the presence of potentially pathogenic strains in seawater in this area. This awareness should be raised among environmental and public health professionals.
Journal of Water and Health
Acanthamoeba spp. is a free-living amoeba that can cause major infections in humans, including keratitis and granulomatous encephalitis. Thus, water resources play an important role in transmitting Acanthamoeba spp. infection to humans. The purpose of this study was to investigate the presence of Acanthamoeba spp. in public swimming pools from three cities of Kerman Province, southeastern Iran. Eighty water samples of 20 public indoor swimming pools were taken from Kerman, Jiroft, and Kahnauj cities. Water temperature (°C), pH, and free chlorine concentration (ppm) were measured. Filtration and cultivation were applied on non-nutrient agar medium. The polymerase chain reaction was applied by using the genus-specific primers (JDP1 and JDP2) on positive samples; these primers can amplify the 423–551 bp fragment. Eighteen of the 20 swimming pools (including 32/80; 40% samples) were contaminated with Acanthamoeba spp. All swimming pools of Jiroft and Kahnauj and 88.2% of swimming pools ...
Journal of Water and Health, 2022
Free-living amoebas (FLAs) can cause neurological and ocular complications in humans. Water supplies play a critical role in transmitting FLAs to humans. The aim of the present study was to investigate the presence of FLAs in various aquatic sources including drinking water, stagnant water, and surface water in Alborz province, northern Iran, using morphological and molecular techniques. A total of 70 water samples were collected from 34 drinking waters, 23 surface waters, and 13 stagnant waters. Filtration and cultivation were employed to isolate FLAs. PCR assay was applied by using the genus-specific primers on positive samples. Pathogenicity tests (osmo-and thermo-tolerance properties) were performed for Acanthamoeba spp., positive sample. Considering the morphological criteria, four positive samples of Acanthamoeba sp., three Vermamoeba sp., two mixed Vermamoeba sp. with Vahlkamfiids, and one mixed Acanthamoeba sp. with Vahlkamfiids were isolated. Five Acanthamoeba sp. isolates were amplified using the JDP primer pairs. Among them, two genotypes, T4 (three isolates) and T5 (two isolates) corresponding to A. lenticulata, were identified. Four V. vermiformis samples were confirmed using the sequencing. This study highlighted the occurrence of potentially pathogenic waterborne FLAs in water habitats associated with high human activity. The results of such research on the prevalence of FLAs, as a human hazard, should be communicated to health policymakers.