scholarly journals High occurrence of Acanthamoeba spp. in the water samples of public swimming pools from Kerman Province, Iran

2021 ◽  
Author(s):  
Raheleh Eftekhari-Kenzerki ◽  
Kavous Solhjoo ◽  
Zahra Babaei ◽  
Hassan Rezanezhad ◽  
Ahmad Abolghazi ◽  
...  
Keyword(s):  
Water Samples ◽  
Agar Medium ◽  
Warning Signs ◽  
Swimming Pools ◽  
Specific Primers ◽  
Free Living ◽  
High Occurrence ◽  
Nutrient Agar Medium ◽  
Free Living Amoeba ◽  
Polymerase Chain

Abstract 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 in Kerman were contaminated. As such, all 32 Acanthamoeba isolates were amplified using the JDP primer pairs. Two genotypes, T3 and T4, were also identified. The present research is the first to report Acanthamoeba spp. in public swimming pools from Kerman Province. Due to high occurrence of this protozoan, it is recommended to use warning signs around swimming pools to create awareness of this infection.

scholarly journals Detection and quantification of human adenovirus genomes in Acanthamoeba isolated from swimming pools

2016 ◽  
Vol 88 (suppl 1) ◽  
pp. 635-641 ◽  
Author(s):  
RODRIGO STAGGEMEIER ◽  
THALITA ARANTES ◽  
KARIN S. CAUMO ◽  
MARILISE B. ROTT ◽  
FERNANDO R. SPILKI
Keyword(s):  
Water Samples ◽  
Human Adenovirus ◽  
Swimming Pools ◽  
Ocular Diseases ◽  
Porto Alegre ◽  
Chain Reaction ◽  
Free Living ◽  
Viral Loads ◽  
Polymerase Chain ◽  
The City

ABSTRACT Acanthamoeba is the most common free-living environmental amoeba, it may serve as an important vehicle for various microorganisms living in the same environment, such as viruses, being pathogenic to humans. This study aimed to detect and quantify human adenoviruses (HAdV) in Acanthamoebas isolated from water samples collected from swimming pools in the city of Porto Alegre, Southern Brazil. Free-living amoebae of the genus Acanthamoeba were isolated from water samples, and isolates (n=16) were used to investigate the occurrence of HAdVs. HAdV detection was performed by quantitative real-time polymerase chain reaction (qPCR). HAdVs were detected in 62.5% (10/16) of Acanthamoeba isolates, ranging from 3.24x103 to 5.14x105 DNA copies per milliliter of isolate. HAdV viral loads found in this study are not negligible, especially because HAdV infections are associated with several human diseases, including gastroenteritis, respiratory distress, and ocular diseases. These findings reinforce the concept that Acanthamoeba may act as a reservoir and promote HAdV transmission through water.

scholarly journals Occurrence of free-living amoebae (Acanthamoeba, Balamuthia, Naegleria) in water samples in Peninsular Malaysia

2018 ◽  
Vol 17 (1) ◽  
pp. 160-171 ◽  
Author(s):  
Shobana Gabriel ◽  
Naveed Ahmed Khan ◽  
Ruqaiyyah Siddiqui
Keyword(s):  
Water Samples ◽  
Tap Water ◽  
Peninsular Malaysia ◽  
Recreational Water ◽  
Chain Reaction ◽  
Specific Primers ◽  
Free Living ◽  
Polymerase Chain ◽  
Centrifugation Method ◽  
Instagene Matrix

Abstract The aim of this study was to determine the occurrence of free-living amoebae (FLA) in Peninsular Malaysia and to compare different methodologies to detect them from water samples. Water samples were collected from tap water, recreational places, water dispensers, filtered water, etc. and tested for FLA using both cultivation and polymerase chain reaction (PCR) via plating assays and centrifugation methods. Amoebae DNA was extracted using Instagene matrix and PCR was performed using genus-specific primers. Of 250 samples, 142 (56.8%) samples were positive for presence of amoebae, while 108 (43.2%) were negative. Recreational water showed higher prevalence of amoebae than tap water. PCR for the plating assays revealed the presence of Acanthamoeba in 91 (64%) samples and Naegleria in 99 (70%) of samples analysed. All samples tested were negative for B. mandrillaris. In contrast, the centrifugation method was less effective in detecting amoebae as only one sample revealed the presence of Acanthamoeba and 52 (29%) samples were positive for Naegleria. PCR assays were specific and sensitive, detecting as few as 10 cells. These findings show the vast distribution and presence of FLA in all 11 states of Peninsular Malaysia. Further studies could determine the possible presence of pathogenic species and strains of free-living amoebae in public water supplies in Malaysia.

scholarly journals Association of Alcaligenes Faecalis Strain in Juvenile Earthworms, from Cocoons of Eudrilus Eugeniae

2019 ◽  
Vol 9 (2S2) ◽  
pp. 688-692
Keyword(s):  
16S Rrna ◽  
Gene Sequence ◽  
Agar Medium ◽  
Alcaligenes Faecalis ◽  
Rrna Gene ◽  
Eudrilus Eugeniae ◽  
Zone Of Inhibition ◽  
Chain Reaction ◽  
Nutrient Agar Medium ◽  
Polymerase Chain

Cocoons of earthworm Eudrilus eugeniae were collected from vermiculture bed and found that it had antibacterial activity. The size of zone of inhibition was directly proportional to the size of cocoons examined. Along with nutritious fluid and embryos, culturable bacterial community was found inside the cocoons. Bacterial colonies were isolated from the trails of newly hatched, juvenile worms in the nutrient agar medium and examined. Gram negative, rod shaped bacterium was found to be abundant in the trails of juvenile earthworms. Polymerase chain reaction was performed from this bacterium to amplify the gene of 16S rRNA and analyzed. Subsequent bi-directional DNA sequencing revealed that this abundant bacterium is highly related to 16S rRNA gene sequence of a strain, Alcaligenes faecalis. Based on available literature, we hypothesize that this bacterium could be symbiotically associated with cocoons of earthworms.

scholarly journals Isolation and identification of Acanthamoeba genotypes and Naegleria spp. from the water samples of public swimming pools in Qazvin, Iran

2019 ◽  
Vol 18 (2) ◽  
pp. 244-251 ◽  
Author(s):  
Nastaran Paknejad ◽  
Elham Hajialilo ◽  
Mehrzad Saraei ◽  
Amir Javadi
Keyword(s):  
Water Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Swimming Pools ◽  
Isolation And Identification ◽  
Highly Pathogenic ◽  
Reaction Amplification ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Naegleria Lovaniensis ◽  
Isolated Species

Abstract Free-living amoeba (FLA), including Acanthamoeba and Naegleria are facultative parasites in humans. The amoeba have widespread distribution in various water sources. The aim of this study was isolation and molecular identification of Acanthamoeba and Naegleria isolated from swimming pools and also hot and cold tub waters in Qazvin province. The samples (166 water samples) were cultured to isolate and identify positive specimens. PCR (polymerase chain reaction) amplification, sequencing and phylogenetic analysis were conducted to confirm the isolated species and genotypes of amoeba. According to morphological characterizations, 18.6% of specimens were identified as FLA, which in 71% were Acanthamoeba by PCR method. Molecular analysis revealed that 36.3%, 18.1% and 4.5% of Acanthamoeba specimens were identified as T3, T4 and T11 Acanthamoeba genotypes, respectively. Protacanthamoeba bohemica (27.2%) and Acanthamoeba sp. (4.5%) were found among the specimens. The results of osmo-tolerance and thermo-tolerance assays demonstrated that 50% of T3 and 25% of T4 genotypes of Acanthamoeba were highly pathogenic parasites. The molecular approach showed the presence of Naegleria lovaniensis (9%) in hot tub water of swimming pools. This study demonstrated that the swimming pools and hot tub water in Qazvin province were contaminated with Acanthamoeba and Naegleria species.

scholarly journals Genotyping of Acanthamoeba spp. Isolated from the Caspian Sea in Iran

2020 ◽  
Vol 2 (1) ◽  
pp. 9
Author(s):  
Mahmoudi M.R. ◽  
Karanis P.
Keyword(s):  
Water Samples ◽  
Caspian Sea ◽  
Nucleotide Sequencing ◽  
Chain Reaction ◽  
The Caspian Sea ◽  
Free Living ◽  
Public Health Professionals ◽  
Causative Agents ◽  
Polymerase Chain ◽  
Seawater Samples

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.

scholarly journals High Occurrence of Potentially Pathogenic Free Living Amoebae in Water Bodies of Kaleybar and Khodaafarin, East Azerbaijan Province

2015 ◽  
Vol 10 (Special-Issue1) ◽  
pp. 727-731
Author(s):  
Hamed Behniafar ◽  
Maryam Niyyati ◽  
Zohreh Lasjerdi ◽  
Samira Dodangeh
Keyword(s):  
Poor Prognosis ◽  
Room Temperature ◽  
Hot Springs ◽  
Agar Medium ◽  
Water Bodies ◽  
Free Living ◽  
Nutrient Agar Medium ◽  
Amebic Encephalitis ◽  
Escherichia Coli Bacteria ◽  
East Azerbaijan

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% non-nutrient 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.

scholarly journals Isolation and molecular identification of free-living amoebae (Naegleria spp., Acanthamoeba spp. and Vermamoeba spp.) from mineral springs in Guilan Province, northern Iran

2020 ◽  
Vol 18 (1) ◽  
pp. 60-66
Author(s):  
Mohammad Hossein Feiz Haddad ◽  
Habib Habibpour ◽  
Mohammad Reza Mahmoudi
Keyword(s):  
Acanthamoeba Castellanii ◽  
Warning Signs ◽  
Nitrocellulose Membrane ◽  
Chain Reaction ◽  
Free Living ◽  
Northern Iran ◽  
Mineral Springs ◽  
Inverted Microscope ◽  
Polymerase Chain ◽  
Guilan Province

Abstract 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 of N. australiensis, two cases of N. pagei and one case of N. gruberi. According to our research the occurrence of Acanthamoeba, Naegleria spp. and Vermamoeba spp. in mineral springs could be hazardous for high risk persons. Regular monitoring and posting warning signs of these waters by health planners could prevent free-living amoebae mediated diseases.

scholarly journals First detection of human sapoviruses in river water in South Africa

2013 ◽  
Vol 67 (12) ◽  
pp. 2776-2783 ◽  
Author(s):  
T. Y. Murray ◽  
J. Mans ◽  
M. B. Taylor
Keyword(s):  
South Africa ◽  
Water Samples ◽  
River System ◽  
Chain Reaction ◽  
Specific Primers ◽  
Potential Health ◽  
Taqman Probes ◽  
Polymerase Chain ◽  
Elution Method ◽  
Three Rivers

Over a 2-year period, from January 2009 to December 2010, water samples were collected from three rivers (Klip, Rietspruit and Suikerbosrand) in the Vaal River System, South Africa. Enteric viruses were recovered by a glass wool adsorption–elution method and concentrated using polyethylene glycol/sodium chloride precipitation. Sapoviruses (SaVs) were detected using published sapovirus (SaV)-specific primers and Taqman probes in a two-step real-time reverse transcription-polymerase chain reaction assay. Based on sequence analysis of the 5′-end of the capsid gene, SaVs were genotyped. In 2009, SaVs were detected in 39% (15/38) of samples from the Klip river, 83% (5/6) from the Rietspruit and 14% (1/7) of samples from the Suikerbosrand river. In 2010, SaVs were detected in 54% (14/26) of Klip river samples, 92% (11/12) from the Rietspruit and 20% (2/10) of samples from the Suikerbosrand river. SaV strains identified in the water samples were characterised into several GI and GII genotypes. The presence of SaVs in these rivers indicates human faecal contamination which may pose a potential health risk to persons exposed to these water sources during domestic or recreational activities.

scholarly journals Co-Occurrence of Free-Living Amoeba and Legionella in Drinking Water Supply Systems

Medicina ◽  
2019 ◽  
Vol 55 (8) ◽  
pp. 492 ◽  
Author(s):  
Valciņa ◽  
Pūle ◽  
Mališevs ◽  
Trofimova ◽  
Makarova ◽  
...  
Keyword(s):  
Drinking Water ◽  
Water Supply ◽  
Water Samples ◽  
Cold Water ◽  
Hot Water ◽  
Drinking Water Supply ◽  
Water Supply Systems ◽  
Free Living ◽  
Free Living Amoeba ◽  
Supply Systems

Background and Objectives: Legionella is one of the most important water-related pathogens. Inside the water supply systems and the biofilms, Legionella interact with other bacteria and free-living amoeba (FLA). Several amoebas may serve as hosts for bacteria in aquatic systems. This study aimed to investigate the co-occurrence of Legionella spp. and FLA in drinking water supply systems. Materials and Methods: A total of 268 water samples were collected from apartment buildings, hotels, and public buildings. Detection of Legionella spp. was performed in accordance with ISO 11731:2017 standard. Three different polymerase chain reaction (PCR) protocols were used to identify FLA. Results: Occurrence of Legionella varied from an average of 12.5% in cold water samples with the most frequent occurrence observed in hot water, in areas receiving untreated groundwater, where 54.0% of the samples were Legionella positive. The occurrence of FLA was significantly higher. On average, 77.2% of samples contained at least one genus of FLA and, depending on the type of sample, the occurrence of FLA could reach 95%. In the samples collected during the study, Legionella was always isolated along with FLA, no samples containing Legionella in the absence of FLA were observed. Conclusions: The data obtained in our study can help to focus on the extensive distribution, close interaction, and long-term persistence of Legionella and FLA. Lack of Legionella risk management plans and control procedures may promote further spread of Legionella in water supply systems. In addition, the high incidence of Legionella-related FLA suggests that traditional monitoring methods may not be sufficient for Legionella control.

Acanthamoeba spp. in river water samples from the Black Sea region, Turkey

2020 ◽  
Vol 18 (2) ◽  
pp. 186-199 ◽  
Author(s):  
İlknur Koyun ◽  
Zeynep Kolören ◽  
Ülkü Karaman ◽  
Amalia Tsiami ◽  
Panagiotis Karanis
Keyword(s):  
Black Sea ◽  
Water Samples ◽  
Tap Water ◽  
Acanthamoeba Keratitis ◽  
The Black Sea ◽  
Free Living ◽  
Black Sea Region ◽  
River Waters ◽  
Free Living Amoeba ◽  
Acanthamoeba Species

Abstract The present study aims to investigate the occurrence of free living amoeba (FLA) in water resources (rivers and tap water) in Samsun in the Black Sea. The presence of Acanthamoeba spp. was confirmed in 98 of 192 water samples collected from 32 sites of Samsun province (Samsun centre, Terme, Carsamba, Tekkekoy, Bafra) by PCR. Acanthamoeba spp. were found in 15/36 river samples from Samsun, in 58/90 from Terme, in 12/30 from Carsamba, in 7/18 from Tekkekoy and in 6/18 from Bafra. No Acanthamoeba species were detected in tap water samples. The highest rate in river waters contaminated with Acanthamoeba species was in Terme followed by Samsun centre (41.7%), Carsamba (40%), Tekkekoy (38.9%) and Bafra districts (33.3%), respectively. The result of the subsequent sequence analysis showed Haplotype I (A. triangularis) in 5%, Haplotype II (A. polyphaga) in 29.6%, Haplotype III (Acanthamoeba spp.) in 62% and Haplotype IV (A. lenticulata) in 3%. The most common genotype was Acanthamoeba T4 (Acanthamoeba spp., A. polyphaga, A. triangularis) and T5 genotype was also found in 3%. The T4 genotype is the most common genotype associated with Acanthamoeba keratitis (AK) worldwide; therefore, humans and animals living in the area are at risk after contact with such waters.

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