scholarly journals Isolation and identification of free-living amoebae isolated from well water in Istanbul

2020 ◽  
Vol 18 (6) ◽  
pp. 1139-1145
Author(s):  
Miray Üstüntürk-Onan
Keyword(s):  
Water Samples ◽  
Pathogenic Bacteria ◽  
Opportunistic Infections ◽  
Health Concern ◽  
Well Water ◽  
Isolation And Identification ◽  
Free Living ◽  
Polymerase Chain ◽  
First Time

Abstract Free-living amoebae (FLA) are ubiquitous protozoa commonly found in water and soil environments. FLA belonging to various genera, including Acanthamoeba, Balamuthia, Naegleria, and Vermamoeba, can cause opportunistic and non-opportunistic infections in humans and animals such as keratitis or meningoencephalitis. In addition, some of them serve as hosts for a large number of pathogenic bacteria, yeasts, and viruses. The purpose of the present study was to assess the prevalence and molecular characterization of FLA in well water samples in İstanbul. Ten well water samples were collected from the taps and the presence of FLA was monitored both by the culture and polymerase chain reaction methods. FLA were isolated in 8 out of the 10 samples (80%) included in this study. Morphological analysis and partial sequencing of the 18S rDNA revealed the presence of Acanthamoeba genotypes T3 and T4, and Vermamoeba vermiformis in the investigated well water samples. This study reports for the first time the detection of Acanthamoeba genotype T3 in well water samples in İstanbul. The presence of potentially pathogenic amoebae in habitats related to human activities supports the relevance of FLA as a potential public health concern.

scholarly journals Prevalence of Protozoa Species in Drinking and Environmental Water Sources in Sudan

2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Salah Shanan ◽  
Hadi Abd ◽  
Magdi Bayoumi ◽  
Amir Saeed ◽  
Gunnar Sandström
Keyword(s):  
Water Samples ◽  
Pathogenic Bacteria ◽  
Molecular Detection ◽  
Environmental Water ◽  
Limited Information ◽  
Free Living ◽  
Detection And Identification ◽  
Animal Pathogens ◽  
Identification Of Species ◽  
Polymerase Chain

Protozoa are eukaryotic cells distributed worldwide in nature and are receiving increasing attention as reservoirs and potential vectors for the transmission of pathogenic bacteria. In the environment, on the other hand, many genera of the protozoa are human and animal pathogens. Only limited information is available on these organisms in developing countries and so far no information on their presence is available from Sudan. It is necessary to establish a molecular identification of species of the protozoa from drinking and environmental water. 600 water samples were collected from five states (Gadarif, Khartoum, Kordofan, Juba, and Wad Madani) in Sudan and analysed by polymerase chain reaction (PCR) and sequencing. 57 out of 600 water samples were PCR positive for protozoa. 38 out of the 57 positive samples were identified by sequencing to contain 66 protozoa species including 19 (28.8%) amoebae, 17 (25.7%) Apicomplexa, 25 (37.9%) ciliates, and 5 (7.6%) flagellates. This study utilized molecular methods identified species belonging to all phyla of protozoa and presented a fast and accurate molecular detection and identification of pathogenic as well as free-living protozoa in water uncovering hazards facing public health.

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 Occurrence and Molecular Characterization of Recombinant Potato virus YNTN Isolates from Sulawesi, Indonesia

Plant Disease ◽  
2016 ◽  
Vol 100 (2) ◽  
pp. 269-275 ◽  
Author(s):  
Mohamad Chikh-Ali ◽  
Nilsa A. Bosque-Pérez ◽  
Dalton Vander Pol ◽  
Dantje Sembel ◽  
Alexander V. Karasev
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Potato Cultivar ◽  
Whole Genome ◽  
Chain Reaction ◽  
Recombinant Type ◽  
Whole Genomes ◽  
Polymerase Chain ◽  
First Time

The importance of potato has increased dramatically in Indonesia over the last three decades. During this period, ‘Granola’, a potato cultivar originally from Germany, has become the most common cultivar for fresh consumption in Indonesia. In August 2014, a survey was conducted in Sulawesi, where potato fields cultivated with Granola and its selection, ‘Super John’, were sampled for Potato virus Y (PVY) presence. PVY was found in Sulawesi for the first time. Samples determined to be positive for PVY were subsequently typed to strain using reverse-transcription polymerase chain reaction assays. All PVY isolates sampled were identified as PVYNTN recombinants, with three recombination junctions in P3, VPg, and CP regions of the genome. Three local PVY isolates were subjected to whole-genome sequencing and subsequent sequence analysis. The whole genomes of the Indonesian PVYNTN isolates I-6, I-16, and I-17 were found to be closely related to the European PVYNTN-A. This recombinant type was shown previously to cause potato tuber necrotic ringspot disease (PTNRD) in susceptible potato cultivars. The dependence of potato farmers on mostly a single cultivar, Granola, may have given a competitive advantage to PVYNTN over other PVY strains, resulting in the predominance of the PVYNTN recombinant. The dominance of PVYNTN in Sulawesi, and possibly in Indonesia as a whole, represents a potential risk to any newly introduced potato cultivar to the country, especially cultivars susceptible to PTNRD.

scholarly journals Genetic Diversity and Aggressiveness of Ralstonia solanacearum Strains Causing Bacterial Wilt of Potato in Uruguay

Plant Disease ◽  
2011 ◽  
Vol 95 (10) ◽  
pp. 1292-1301 ◽  
Author(s):  
M. I. Siri ◽  
A. Sanabria ◽  
M. J. Pianzzola
Keyword(s):  
Genetic Diversity ◽  
Bacterial Wilt ◽  
Ralstonia Solanacearum ◽  
Potato Tubers ◽  
Bacterial Strains ◽  
Potato Plants ◽  
Polymerase Chain ◽  
First Time ◽  
Different Levels

Bacterial wilt, caused by Ralstonia solanacearum, is a major disease affecting potato (Solanum tuberosum) production worldwide. Although local reports suggest that the disease is widespread in Uruguay, characterization of prevalent R. solanacearum strains in that country has not been done. In all, 28 strains of R. solanacearum isolated from major potato-growing areas in Uruguay were evaluated, including 26 strains isolated from potato tubers and 2 from soil samples. All strains belonged to phylotype IIB, sequevar 1 (race 3, biovar 2). Genetic diversity of strains was assessed by repetitive-sequence polymerase chain reaction, which showed that the Uruguayan strains constituted a homogeneous group. In contrast, inoculation of the strains on tomato and potato plants showed, for the first time, different levels of aggressiveness among R. solanacearum strains belonging to phylotype IIB, sequevar 1. Aggressiveness assays were also performed on accessions of S. commersonii, a wild species native to Uruguay that is a source of resistance for potato breeding. No significant interactions were found between bacterial strains and potato and S. commersonii genotypes, and differences in aggressiveness among R. solanacearum strains were consistent with previously identified groups based on tomato and potato inoculations. Moreover, variation in responses to R. solanacearum was observed among the S. commersonii accessions tested.

scholarly journals Recovery of Salmonella from biofilms in a headwater spring ecosystem

2011 ◽  
Vol 9 (3) ◽  
pp. 458-466 ◽  
Author(s):  
James P. Gaertner ◽  
Joseph A. Mendoza ◽  
Michael R. J. Forstner ◽  
Dittmar Hahn
Keyword(s):  
Pathogenic Bacteria ◽  
Chain Reaction ◽  
Selective Enrichment ◽  
Animal Wastes ◽  
San Marcos ◽  
Polymerase Chain ◽  
Spring Lake ◽  
Precipitation Events

Salmonellae are pathogenic bacteria often detected in waters impacted by human or animal wastes. In order to assess the fate of salmonellae in supposedly pristine environments, water and natural biofilm samples along with snails (Tarebia granifera) and crayfish (Procambarus clarkia) were collected before and up to 7 days following four precipitation events from sites within the headwater springs of Spring Lake, San Marcos, TX. The samples were analyzed for the presence of salmonellae by polymerase chain reaction (PCR) after semi-selective enrichment. Salmonellae were detected in one water sample directly after precipitation only, while detection in ten biofilm and two crayfish samples was not related to precipitation. Salmonellae were not detected in snails. Characterization of isolates by rep-PCR revealed shared profiles in water and biofilm samples, biofilm and crayfish samples, and biofilm samples collected 23 days apart. These results suggest that salmonellae are infrequently washed into this aquatic ecosystem during precipitation runoff and can potentially take up residency in biofilms which can help facilitate subsequent long-term persistence and eventual transfer through the food chain.

Potentially pathogenic bacteria isolated from different tropical waters in Sri Lanka

2013 ◽  
Vol 13 (6) ◽  
pp. 1463-1469 ◽  
Author(s):  
W. M. G. C. K. Mannapperuma ◽  
C. L. Abayasekara ◽  
G. B. B. Herath ◽  
D. R. I. B. Werellagama
Keyword(s):  
Surface Water ◽  
Sri Lanka ◽  
Water Samples ◽  
Pathogenic Bacteria ◽  
Water Sources ◽  
Bottled Water ◽  
Well Water ◽  
Faecal Coliforms ◽  
Total Coliforms ◽  
Source Waters

This study investigated the bacteriological contamination of different water sources in Sri Lanka. Source waters (n = 74) including bottled water, well water and surface water were assessed for enumerating total coliforms and faecal coliforms using the membrane filtration method. The results showed that 18.5 and 14.8% of bottled water samples were contaminated with total coliforms and faecal coliforms, respectively. All the well water and surface water samples exceeded the WHO permitted levels for total coliforms and faecal coliforms. Bacteriological identification using biochemical tests and api 20E identification tests revealed the presence of potentially pathogenic bacteria in all water sources tested. Bottled water mainly contained three Enterobacter species, while well water samples showed the broadest spectrum of bacteria including eleven coliform species belonging to the genera Klebsiella, Escherichia, Enterobacter, Citrobacter, Kluyvera, Pantoea, Rautella, and 10 non-coliform species in the genera Pseudomonas, Aeromonas, Salmonella and Acinetobacter. Surface waters contained seven coliform species belonging to the genera Klebsiella, Escherichia, Citrobacter, Rautella and Serratia, and eight non-coliform species in the genera Pseudomonas, Aeromonas and Acinetobacter. Detection of higher bacteriological counts and identification of potentially pathogenic bacteria in different source waters suggest a potential health risk of the water sources used in Sri Lanka.

scholarly journals Isolation and molecular characterization of Acanthamoeba genotypes in recreational and domestic water sources from Jamaica, West Indies

2015 ◽  
Vol 13 (3) ◽  
pp. 909-919 ◽  
Author(s):  
Cheridah D. Todd ◽  
María Reyes-Batlle ◽  
José E. Piñero ◽  
Enrique Martínez-Carretero ◽  
Basilio Valladares ◽  
...  
Keyword(s):  
Water Sources ◽  
Skin Infections ◽  
Domestic Water ◽  
Free Living ◽  
Water Contact ◽  
First Report ◽  
The Caribbean ◽  
Polymerase Chain ◽  
Neurological Infections

Free living amoebae (FLA) are amphizoic protozoa that are ubiquitous in nature. Infection with FLA may result in neurological, ocular and skin infections. Exposure to Acanthamoeba occurs frequently through water contact and knowledge of the presence of the organisms in water sources is important in understanding transmission dynamics. The distribution of Acanthamoeba was studied in recreational and domestic water samples collected from across Jamaica. Morphological assessment and polymerase chain reaction revealed Acanthamoeba spp. isolates in 50.6% (42/83) and 17.3% (14/81) of recreational and domestic water, respectively. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotypes T3, T4, T5, T10 and T11 corresponding to Acanthamoeba spp: A. griffini, A. triangularis, A. lenticulata, A. culbertsoni and A. hatchetti. Moreover, T4 was the most frequently isolated genotype in both recreational and domestic water. Thermotolerance and osmotolerance assays indicated that most isolates were potentially pathogenic. This is the first report of T3 and T10 genotypes in the Caribbean and the first report of these Acanthamoeba spp. in Jamaican waters. The study shows that there is potential risk of infection to contact wearers who practise poor lens care. Further, Acanthamoeba should be considered as a cause of neurological infections in Jamaica.

Evaluation of methods for isolation of DNA for polymerase chain reaction (PCR)-based identification of pathogenic bacteria from pure cultures and water samples

2008 ◽  
Vol 58 (5) ◽  
pp. 995-999 ◽  
Author(s):  
K. Horáková ◽  
H. Mlejnková ◽  
P. Mlejnek
Keyword(s):  
Polymerase Chain Reaction ◽  
Water Samples ◽  
Pathogenic Bacteria ◽  
Pcr Amplification ◽  
Bacterial Suspension ◽  
Chain Reaction ◽  
Alkaline Lysis ◽  
Bacterial Dna ◽  
Pure Cultures ◽  
Polymerase Chain

Polymerase chain reaction (PCR) provides a reliable detection of pathogenic bacteria in water samples. However, this method can be adversely influenced by the purity of the DNA template. This is a particularly important obstacle when the bacterial DNA is directly extracted from water samples. In this study we compared the suitability of 8 different methods for isolation of bacterial DNA from pure cultures and 10 different methods for isolation of DNA from water samples. The quality of extracted DNA was assessed by PCR amplification of target sequences derived from uid (E. coli and Shigella sp.), tuf (Enterococcus sp.) and hns (Salmonella sp.). Results indicated that there are differences among the methods tested and only a few of them gave satisfactory results. The method based on alkaline lysis of bacterial suspension, which was developed in our laboratory, seemed to be efficient enough for the detection of bacteria from pure cultures. Detection of bacteria directly from water samples was more difficult. The modified method developed by Slusarenko was found as the best of the tested methods.

scholarly journals Co-Existence of Free-Living Amoebae and Potential Human Pathogenic Bacteria Isolated from Rural Household Water Storage Containers

Biology ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1228
Author(s):  
Natasha Potgieter ◽  
Clarissa van der Loo ◽  
Tobias George Barnard
Keyword(s):  
Rural Communities ◽  
Water Samples ◽  
Pathogenic Bacteria ◽  
Water Source ◽  
Culture Methods ◽  
Water Shortages ◽  
Free Living ◽  
Human Pathogenic Bacteria ◽  
Rrna Sequencing ◽  
Primary Water

This study investigated the co-existence of potential human pathogenic bacteria and free-living amoebae in samples collected from stored water in rural households in South Africa using borehole water as a primary water source. Over a period of 5 months, a total of 398 stored water and 392 biofilm samples were collected and assessed. Free-living amoebae were identified microscopically in 92.0% of the water samples and 89.8% of the biofilm samples. A further molecular identification using 18S rRNA sequencing identified Vermamoeba vermiformis, Entamoeba spp., Stenamoeba spp., Flamella spp., and Acanthamoeba spp. including Acanthamoeba genotype T4, which is known to be potentially harmful to humans. Targeted potential pathogenic bacteria were isolated from the water samples using standard culture methods and identified using 16S rRNA sequencing. Mycobacterium spp., Pseudomonas spp., Enterobacter spp., and other emerging opportunistic pathogens such as Stenotrophomonas maltophilia were identified. The results showed the importance of further studies to assess the health risk of free-living amoebae and potential human pathogenic bacteria to people living in rural communities who have no other option than to store water in their homes due to water shortages.

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.

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