A rapid two dot filter assay for the detection of E. coli O157 in water samples

2008 ◽  
Vol 336 (2) ◽  
pp. 159-165 ◽  
Author(s):  
Sujatha Kamma ◽  
Lily Tang ◽  
Kelvin Leung ◽  
Edie Ashton ◽  
Norman Newman ◽  
...  
Keyword(s):  
Water Samples ◽  
E Coli ◽  
Filter Assay

Campylobacter in waterborne epidemics in Finland

2004 ◽  
Vol 4 (2) ◽  
pp. 39-45 ◽  
Author(s):  
M.-L. Hänninen ◽  
R. Kärenlampi
Keyword(s):  
Campylobacter Jejuni ◽  
Causative Agent ◽  
Water Samples ◽  
Fecal Coliforms ◽  
Risk Level ◽  
Source Water ◽  
Indicator Organisms ◽  
Faecal Contamination ◽  
E Coli ◽  
July August September

The sources for drinking water in Finland are surface water, groundwater or artificially recharged groundwater. There are approximately 1400 groundwater plants in Finland that are microbiologically at a high risk level because in most cases they do not use any disinfection treatment. Campylobacter jejuni has caused waterborne epidemics in several countries. Since the middle of the 1980s, C. jejuni has been identified as the causative agent in several waterborne outbreaks in Finland. Between 1998 and 2001, C. jejuni or C. upsaliensis caused seven reported waterborne epidemics. In these epidemics approximately 4000 people acquired the illness. Most of the outbreaks occurred in July, August , September or October. In four of them source water and net water samples were analysed for total coliforms or fecal coliforms, E. coli and campylobacters. We showed that large volumes of water samples in studies of indicator organisms (up to 5000 ml) and campylobacters (4000–20,000 ml) increased the possibility to identify faecal contamination and to detect the causative agent from suspected sources.

An Improved Membrane Filtration Method for Enumeration of Faecal Coliforms and E. Coli by a Fluorogenic β-Glucuronidase Assay

1993 ◽  
Vol 27 (3-4) ◽  
pp. 267-270 ◽  
Author(s):  
M. T. Augoustinos ◽  
N. A. Grabow ◽  
B. Genthe ◽  
R. Kfir
Keyword(s):  
Escherichia Coli ◽  
Water Samples ◽  
Membrane Filtration ◽  
Faecal Coliforms ◽  
Environmental Waters ◽  
Filtration Method ◽  
E Coli ◽  
Wide Range ◽  
Pure Cultures ◽  
Glucuronidase Assay

A fluorogenic β-glucuronidase assay comprising membrane filtration followed by selective enumeration on m-FC agar at 44.5°C and further confirmation using tlie 4-metliylumbelliferyl-β-D-glucuronide (MUG) containing medium was evaluated for the detection of Escherichia coli in water. A total of 200 typical blue and non-typical blue colonies were isolated from sea and fresh water samples using initial selective enumeration on m-FC agar. Pure cultures of the selected colonies were further tested using the MUG assay and identified using the API 20E method. Of the colonies tested which were shown to be positive using the MUG assay 99.4% were Escherichia coli. The results of this study indicate the combination of the m-FC method followed by the MUG assay to be highly efficient for the selection and confirmation of E. coli from a wide range of environmental waters.

Occurrences of Giardia cysts in beach water

1998 ◽  
Vol 38 (12) ◽  
pp. 73-76 ◽  
Author(s):  
B. S. W. Ho ◽  
T.-Y Tam
Keyword(s):  
Health Risk ◽  
Water Sample ◽  
Water Samples ◽  
High Concentration ◽  
Potential Health ◽  
Infectious Dose ◽  
Potential Health Risk ◽  
E Coli ◽  
Beach Water ◽  
Giardia Cysts

A total of 64 beach water samples with various bacteriological quality (Grades 1 to 4) were analysed for their bacteriological and parasitological contents (E coli and Giardia cysts respectively). Results indicated that Giardia cysts were detected in less than 10% of the Grade 1 beach water samples with E coli concentrations of <24/100mL. For Grades 2, 3 & 4 beach water samples, Giardia cysts were found, respectively, in 85, 50 and 64% of the samples. Except for one beach water sample which had an unusually high concentration of Giardia cysts (23 cysts/L), they were generally present at moderate concentrations (<10 cysts/L) in all other beach water samples. Despite moderate levels of Giardia cysts present in beach water of different grades, the potential health risk faced by swimmers bathing in local beach water needs to be carefully assessed as Giardia is known to have a low infectious dose.

scholarly journals Microbiological evaluation of lettuce produced by conventional and organic systems in farms of Londrina, PR

2017 ◽  
Vol 38 (1) ◽  
pp. 175 ◽  
Author(s):  
Natalia Harumi Niguma ◽  
Jacinta Sanchez Pelayo ◽  
Tereza Cristina Rocha Moreira de Oliveira
Keyword(s):  
Irrigation Water ◽  
Water Samples ◽  
Total Coliform ◽  
Organic Production ◽  
Conventional System ◽  
Salmonella Spp ◽  
Organic System ◽  
E Coli ◽  
Organic Systems ◽  
Production Practices

The aims of this study were to evaluate the contamination of lettuce (Lactuca sativa), produced in Londrina, Paraná (PR), with total coliform, coliform at 45 °C, E. coli, and Salmonella spp.; and to determine the E. coli contamination of irrigation water used at the farms studied. Four farms were evaluated, of which three produced lettuce using a conventional system and one using an organic system. An evaluation of the production practices of the farms was also carried out. A total of 111 samples were analyzed, 71 lettuce samples from the conventional system and 40 samples from the organic system. A total of eight irrigation water samples were collected for analysis. Coliform at 45 °C counts above the limit tolerated by Brazilian legislation were observed in 2.8% (2/71) of conventionally grown lettuce samples, and Salmonella spp. was isolated in 1.4% (1/71) of those samples. In the organic lettuce samples, 12.5% (5/40) had coliform at 45 °C counts above the limit tolerated and Salmonella spp. was not detected. Irrigation water samples from three farms were unsatisfactory, with counts higher than 102MPN of E. coli per 100mL. The results of this study demonstrate that most conventionally grown lettuce samples show good sanitary conditions in production, and that lettuce contamination is not related to contamination found in irrigation water samples. The results also showed that the organic production practices required by Brazilian certification agencies should be applied to ensure that contamination of produced lettuce remains controlled.

scholarly journals Development of an Internal Control for Evaluation and Standardization of a Quantitative PCR Assay for Detection of Helicobacter pylori in Drinking Water

2007 ◽  
Vol 73 (22) ◽  
pp. 7380-7387 ◽  
Author(s):  
Keya Sen ◽  
Nancy A. Schable ◽  
Dennis J. Lye
Keyword(s):  
Helicobacter Pylori ◽  
Drinking Water ◽  
Quantitative Pcr ◽  
Water Samples ◽  
Sodium Thiosulfate ◽  
Qpcr Assay ◽  
E Coli ◽  
H Pylori ◽  
Labeled Probe ◽  
Treated Drinking Water

ABSTRACT Due to metabolic and morphological changes that can prevent Helicobacter pylori cells in water from growing on conventional media, an H. pylori-specific TaqMan quantitative PCR (qPCR) assay was developed that uses a 6-carboxyfluorescein-labeled probe (A. E. McDaniels, L. Wymer, C. Rankin, and R. Haugland, Water Res. 39:4808-4816, 2005). However, proper internal controls are needed to provide an accurate estimate of low numbers of H. pylori in drinking water. In this study, the 135-bp amplicon described by McDaniels et al. was modified at the probe binding region, using PCR mutagenesis. The fragment was incorporated into a single-copy plasmid to serve as a PCR-positive control and cloned into Escherichia coli to serve as a matrix spike. It was shown to have a detection limit of five copies, using a VIC dye-labeled probe. A DNA extraction kit was optimized that allowed sampling of an entire liter of water. Water samples spiked with the recombinant E. coli cells were shown to behave like H. pylori cells in the qPCR assay. The recombinant E. coli cells were optimized to be used at 10 cells/liter of water, where they were shown not to compete with 5 to 3,000 cells of H. pylori in a duplex qPCR assay. Four treated drinking water samples spiked with H. pylori (100 cells) demonstrated similar cycle threshold values if the chlorine disinfectant was first neutralized by sodium thiosulfate.

scholarly journals Detection of antimicrobial resistance genes of carbapenem-resistant Enterobacteriaceae in Escherichia coli isolated from the water supply of smallholder dairy farms in Saraburi and Maha Sarakham, Thailand

2020 ◽  
Vol 6 (1) ◽  
pp. 1-5
Author(s):  
Natapol Pumipuntu ◽  
Sangkom Pumipuntu
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Water Samples ◽  
Dairy Farms ◽  
Drug Resistant ◽  
E Coli ◽  
Carbapenem Resistant ◽  
Carbapenem Resistant Enterobacteriaceae ◽  
Smallholder Dairy Farms

Background and Aim: The problem of antimicrobial resistance of bacteria in both humans and animals is an important public health concern globally, which is likely to increase, including in Thailand, where carbapenem-resistant Enterobacteriaceae (CRE), such as Escherichia coli, are of particular concern. They are pathogens found in the gastrointestinal tract of humans and other animals as well as in the environment. They may cause opportunistic infection and are often resistant to antibiotics in various fields especially in animal husbandry, such as pets or livestock farms. This study aimed to investigate the occurrence of carbapenem-resistant E. coli from water samples of smallholder dairy farms in Saraburi and Maha Sarakham, Thailand. Materials and Methods: Sixty-four water samples were collected from 32 dairy farms in Kaeng Khoi district, Muak Lek district, and Wang Muang district of Saraburi Province, and Kantharawichai district and Mueang district of Maha Sarakham Province, Thailand. All samples were cultured and isolated for E. coli by biochemical tests. All E. coli isolates were tested for drug susceptibility using imipenem, meropenem, and drug resistance genes of carbapenemases such as blaNDM, blaIMP, and blaOXA48 of drug-resistant E. coli isolates detected by polymerase chain reaction (PCR) technique. Results: A total of 182 E. coli isolates were found (140 and 42 isolates from Saraburi and Maha Sarakham, respectively). Drug sensitivity tests found that two isolates of E. coli from water in Kaeng Khoi were resistant to imipenem; therefore, the incidence of E. coli resistance to carbapenem was 1.43% of Saraburi Province. On the other hand, there was no incidence of drug-resistant E. coli in Maha Sarakham. In addition, the detection of the drug-resistant gene of E. coli in both isolates by PCR showed the expression of blaNDM. Conclusion: This study reports E. coli resistance to antimicrobial drugs on livestock farms. It can be considered to be the first report of E. coli CRE detection in a dairy farm at Saraburi, which should be the subject of further extended study.

scholarly journals Distribution and Differential Survival of Traditional and Alternative Indicators of Fecal Pollution at Freshwater Beaches

2016 ◽  
Vol 83 (4) ◽  
Author(s):  
Danielle D. Cloutier ◽  
Sandra L. McLellan
Keyword(s):  
Water Samples ◽  
Environmental Samples ◽  
High Sensitivity ◽  
Fecal Pollution ◽  
Source Material ◽  
Beach Sand ◽  
Differential Survival ◽  
Content Type ◽  
Volume Comparison ◽  
E Coli

ABSTRACT Alternative indicators have been developed that can be used to identify host sources of fecal pollution, yet little is known about how their distribution and fate compare to traditional indicators. Escherichia coli and enterococci were widely distributed at the six beaches studied and were detected in almost 95% of water samples (n = 422) and 100% of sand samples (n = 400). Berm sand contained the largest amount of E. coli (P < 0.01), whereas levels of enterococci were highest in the backshore (P < 0.01). E. coli and enterococci were the lowest in water, using a weight-to-volume comparison. The gull-associated Catellicoccus marimammalium (Gull2) marker was found in over 80% of water samples, regardless of E. coli levels, and in 25% of sand samples. Human-associated Bacteroides (HB) and Lachnospiraceae (Lachno2) were detected in only 2.4% of water samples collected under baseflow and post-rain conditions but produced a robust signal after a combined sewage overflow, despite low E. coli concentrations. Burdens of E. coli and enterococci in water and sand were disproportionately high in relation to alternative indicators when comparing environmental samples to source material. In microcosm studies, Gull2, HB, and Lachno2 quantitative PCR (qPCR) signals were reduced twice as quickly as those from E. coli and enterococci and approximately 20% faster than signals from culturable E. coli. High concentrations of alternative indicators in source material illustrated their high sensitivity for the identification of fecal sources; however, differential survival and the potential for long-term persistence of traditional fecal indicators complicate the use of alternative indicator data to account for the levels of E. coli and enterococci in environmental samples. IMPORTANCE E. coli and enterococci are general indicators of fecal pollution and may persist in beach sand, making their use problematic for many applications. This study demonstrates that gull fecal pollution is widespread at Great Lakes beaches, whereas human and ruminant contamination is evident only after major rain events. An exploration of sand as a reservoir for indicators found that E. coli was ubiquitous, while gull host markers were detected in only 25% of samples. In situ sand beach microcosms provided decay rate constants for E. coli and enterococci relative to alternative indicators, which establish comparative benchmarks that would be helpful to distinguish recent from past pollution. Overall, alternative indicators are useful for identifying sources and assessing potentially high health risk contamination events; however, beach managers should be cautious in attempting to directly link their detection to the levels of E. coli or enterococci.

scholarly journals Isolation and Enumeration of Escherichia coli from Soil and Water

2020 ◽  
Vol 36 (2) ◽  
pp. 75-77
Author(s):  
Anindita Bhowmik ◽  
Sunjukta Ahsan
Keyword(s):  
Escherichia Coli ◽  
Water Samples ◽  
Gas Production ◽  
Soil Samples ◽  
Most Probable Number ◽  
Biochemical Tests ◽  
Probable Number ◽  
E Coli ◽  
Dilution Series

Majority of the population of Bangladesh depend on tap or surface water as their source of water supply. This study was carried out to examine the microbial quality of both water and soil collected from different places using the multiple tube fermentation technique to determine coliform count by the most probable number (MPN) method in brilliant green lactose broth (BGLB) media.Inoculum from positive tubes of the presumptive test were further transferred on eosinemethylene blue (EMB) and MacConkey agar.The organisms isolated were further characterized using biochemical tests. Out of 93 water samples, 30 (32.26%) indicated the presence of lactose fermenter and gas producer in all 3 tubes of dilution series using inoculum quantities of 1.0, 0.1 and 0.01 ml, whereas out of 85 soil samples, 45 (52.94%) showed acid and gas production in all 3 tubes of dilution series.Among 85 soil samples, 40 samples that contained at least one positive in each dilution series and among 93 water samples, 31 samples that contained at least one positive in each dilution series were further re-identified with biochemical tests.This study showed 30.59% soil isolates and 26.88% water isolates were Escherichia coli which highlighted the fact that both water and soil act as a major reservoir of E.coli, which indicates possible fecal contamination as well as presence of potentially pathogenic E. coli. Bangladesh J Microbiol, Volume 36 Number 2 December 2019, pp 75-77

scholarly journals Borehole water: a potential health risk to rural communities in South Africa

2018 ◽  
Vol 19 (1) ◽  
pp. 128-136 ◽  
Author(s):  
S. Taonameso ◽  
L. S. Mudau ◽  
A. N. Traoré ◽  
N. Potgieter
Keyword(s):  
Drinking Water ◽  
Rural Communities ◽  
Water Samples ◽  
Rural Areas ◽  
Total Coliform ◽  
Water Sources ◽  
Potential Health ◽  
Point Mapping ◽  
E Coli ◽  
Mapping Tool

Abstract Sporadic outbreaks of diarrhoea in children in the Vhembe rural areas could be an indication of contamination in drinking water sources. In areas where improved water sources are used, not all rural households experience the benefits of these improved water sources. Water samples were collected from boreholes in three wards in the Vhembe District to determine microbiological risks over a 5-month period. A Water Point Mapping tool was used to indicate the borehole distribution. Water samples were taken from each functional borehole and analysed for total coliform and Escherichia coli counts, electrical conductivity, pH and temperature. A multiplex PCR protocol was used for identification of pathogenic E. coli. A total of 125 boreholes were identified of which only 12 were functional. Seven boreholes tested positive for total coliforms and E. coli counts. Four boreholes (33.3%) tested positive for diarrhoeagenic E. coli. Fifty-eight percent (58%) of water samples were without health risks, 17% were low risk and 25% could cause infection according to the South African water quality standards. This study indicated the importance of the role of the Municipalities and the maintenance plans that need to ensure that all boreholes are functional and provide safe drinking water to the rural communities.

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