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 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.

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.

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 Critical points and the presence of pathogenic bacteria in iced beverage processing lines

2017 ◽  
Vol 11 (06) ◽  
pp. 493-500
Author(s):  
Halim Nababan ◽  
Winiati Pudji Rahayu ◽  
Diana Elizabeth Waturangi ◽  
Suratmono Suratmono ◽  
Rina Puspitasari ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Critical Points ◽  
Water Samples ◽  
Pathogenic Bacteria ◽  
Control Point ◽  
Enterotoxigenic Escherichia Coli ◽  
Critical Control Point ◽  
Production And Distribution ◽  
Polymerase Chain ◽  
Ice Production

Introduction: Ice can be contaminated by pathogenic bacteria. This study aimed to identify critical points in iced beverage production and distribution lines to examine the presence of pathogenic bacteria in a beverage and its processing environment, as well as when water and ice used as main ingredients. Methodology: The critical points were determined using the principles of Hazard Analytical Critical Control Point (HACCP) to analyze each processing and distribution step from the survey. Samples collected from the points of concern based on the critical points that were found were tested for pathogens by conventional method and molecular method using primers and polymerase chain reaction (PCR). Results: Escherichia coli was found in 6.34% of samples, and 0.7% of them were confirmed as enterotoxigenic Escherichia coli (ETEC) by PCR. Vibrio cholerae was found in in 0.7% of water samples used to make iced beverages and in ice production, as well as in 2.12% of distribution and production tools. Salmonella Typhimurium was found in 1.4% of water samples used to make ice and ice products. Staphylococcus aureus was found in 2.02% of the surfaces of ice distribution and production tools and in 5.05% of production and distribution workers’ hands. S. aureus counts ranged from 2.4×102 - 3.5×102 colonies/100 cm² surface area and 1.9×10¹ - 3.7×102 colonies/workers’ hands. Conclusion: Control on many critical points in iced beverage processing and distribution is required so that the beverages are safe for consumption.

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 Élelmiszer-biztonsági jelentőségű baktériumok molekuláris detektálásának fejlesztése miniatürizált mikrofluidikai eszközök segítségével

2015 ◽  
Vol 156 (51) ◽  
pp. 2082-2088
Author(s):  
Kristóf Iván ◽  
Anna Maráz
Keyword(s):  
Polymerase Chain Reaction ◽  
Pathogenic Bacteria ◽  
Lab On A Chip ◽  
Dna Amplification ◽  
Molecular Techniques ◽  
Microbiological Analysis ◽  
Chain Reaction ◽  
Detection And Identification ◽  
Food Borne ◽  
Polymerase Chain

Detection and identification of food-borne pathogenic bacteria are key points for the assurance of microbiological food safety. Traditional culture-based methods are more and more replaced by or supplemented with nucleic acid based molecular techniques, targeting specific (preferably virulence) genes in the genomes. Internationally validated DNA amplification – most frequently real-time polymerase chain reaction – methods are applied by the food microbiological testing laboratories for routine analysis, which will result not only in shortening the time for results but they also improve the performance characteristics (e.g. sensitivity, specificity) of the methods. Beside numerous advantages of the polymerase chain reaction based techniques for routine microbiological analysis certain drawbacks have to be mentioned, such as the high cost of the equipment and reagents, as well as the risk of contamination of the laboratory environment by the polymerase chain reaction amplicons, which require construction of an isolated laboratory system. Lab-on-a-chip systems can integrate most of these laboratory processes within a miniaturized device that delivers the same specificity and reliability as the standard protocols. The benefits of miniaturized devices are: simple – often automated – use, small overall size, portability, sterility due to single use possibility. These miniaturized rapid diagnostic tests are being researched and developed at the best research centers around the globe implementing various sample preparation and molecular DNA amplification methods on-chip. In parallel, the aim of the authors’ research is to develop microfluidic Lab-on-a-chip devices for the detection and identification of food-borne pathogenic bacteria. Orv. Hetil., 2015, 156(51), 2082–2088.

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