scholarly journals Membrane filtration media for the enumeration of coliform organisms and Escherichia coli in water: comparison of Tergitol 7 and lauryl sulphate with Teepol 610

1980 ◽  
Vol 85 (2) ◽  
pp. 181-191 ◽  
Author(s):  
Keyword(s):  
Escherichia Coli ◽  
Water Samples ◽  
False Positive ◽  
Membrane Filtration ◽  
Sodium Lauryl Sulphate ◽  
Standard Medium ◽  
E Coli ◽  
Laboratory Trial ◽  
Positive Results ◽  
Filtration Technique

SUMMARYIn a multi-laboratory trial with the mombrane filtration technique, three surfactants – Teepol 610 (T610), Tergitol 7 (T7) and sodium lauryl sulphate (LS) – were compared in media for the enumeration of coliform organisms and Escherichia coli in water. A total of 170 samples of water (87 raw and 92 marginally chlorinated) were examined for colony counts of coliform organisms, and 185 water samples (94 raw and 91 marginally chlorinated) for E. coli. Slight differences in the confirmed colony counts between the three media were noted, but few of these were observed consistently in every laboratory. In most laboratories, T7 gave slightly higher counts of E. coli than LS with chlorinated waters; a higher incidence of false-positive results for E. coli at 44 °C was also noted with T7. As there were no outstanding differences in the trial, sodium lauryl sulphate, which is chemically defined, cheap and readily available, is therefore recommended for use at a concentration of 0·1% instead of Teepol 610 in the standard medium for the enumeration of coliform organisms and E. coli in water by the membrane filtration technique.

scholarly journals Marine Bacteria Cause False-Positive Results in the Colilert-18 Rapid Identification Test for Escherichia coli in Florida Waters

2002 ◽  
Vol 68 (2) ◽  
pp. 539-544 ◽  
Author(s):  
John M. Pisciotta ◽  
Damon F. Rath ◽  
Paul A. Stanek ◽  
D. Michael Flanery ◽  
Valerie J. Harwood
Keyword(s):  
Escherichia Coli ◽  
False Positive ◽  
Fecal Coliform ◽  
Membrane Filtration ◽  
False Positives ◽  
Fecal Coliforms ◽  
Marine Waters ◽  
E Coli ◽  
Marine Samples ◽  
Positive Results

ABSTRACT The Colilert-18 system for enumeration of total coliforms and Escherichia coli is approved by the U.S. Environmental Protection Agency for use in drinking water analysis and is also used by various agencies and research studies for enumeration of indicator organisms in fresh and saline waters. During monitoring of Pinellas County, Fla., marine waters, estimates of E. coli numbers (by Colilert-18) frequently exceeded fecal coliform counts (by membrane filtration) by 1 to 3 orders of magnitude. Samples from freshwater sites did not display similar discrepancies. Fecal coliforms, including E. coli, could be cultured from 100% of yellow fluorescent wells (denoting E. coli-positive results) inoculated with freshwater samples but could be cultured from only 17.1% of the “positive” wells inoculated with marine samples. Ortho-nitrophenyl-β-d-galactopyranoside (ONPG)-positive or 4-methylumbelliferyl-β-d-glucuronide (MUG)-positive noncoliform bacteria were readily cultured from Colilert-18 test wells inoculated with marine samples. Filtered cell-free seawater did not cause false positives. Coculture preparations of as few as 5 CFU of Vibrio cholerae (ONPG positive) and Providencia sp. (MUG positive) ml−1 inoculated into Colilert-18 caused false-positive E. coli results. Salinity conditions influenced coculture results, as the concentration of coculture inoculum required to cause false positives in most wells increased from about 5 CFU ml−1 in seawater diluted 1:10 with freshwater to ≈5,000 CFU ml−1 in seawater diluted 1:20 with freshwater. Estimated E. coli numbers in various marine water samples processed at the 1:10 dilution ranged from 10 to 7,270 CFU�100 ml−1, while E. coli numbers in the same samples processed at the 1:20 dilution did not exceed 40 CFU�100 ml−1. The lower estimates of E. coli numbers corresponded well with fecal coliform counts by membrane filtration. This study indicates that assessment of E. coli in subtropical marine waters by Colilert-18 is not accurate when the recommended 1:10 sample dilution is used. The results suggest that greater dilution may diminish the false-positive problem, but further study of this possibility is recommended.

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.

Comparison of three different media for the detection of E. coli and coliforms in water

2006 ◽  
Vol 54 (3) ◽  
pp. 141-145 ◽  
Author(s):  
C. Bernasconi ◽  
G. Volponi ◽  
L. Bonadonna
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Membrane Filtration ◽  
Simultaneous Detection ◽  
Alternative Methods ◽  
Phenotypic Characteristic ◽  
Iso Standard ◽  
E Coli ◽  
Microbiological Parameters ◽  
Filtration Technique

The European Drinking Water Directive defines reference methods for the enumeration of microbiological parameters in drinking water. The method to be used for Escherichia coli and coliforms is the membrane filtration technique on Lactose TTC agar with Tergitol 7. Many technical drawbacks of the procedure, as well as its limitations regarding the recent taxonomy of coliforms, make it necessary to evaluate alternative methods. Two alternative assays, a chromogenic media (m-ColiBlu24®) and a defined substrate technology-DST test (Colilert 18/Quanty Tray™) were compared with the ISO standard with attention to the phenotypic characteristic of the isolates. Results showed that the ISO method failed to detect an important percentage of coliforms and E. coli while m-ColiBlu24® and Colilert 18 provided results in a shorter time allowing the simultaneous detection of E. coli and coliforms with no further confirmation steps.

scholarly journals Detection of AmpC β-lactamases in Escherichia coli using different screening agars

2019 ◽  
Author(s):  
Evert den Drijver ◽  
Jaco J. Verweij ◽  
Carlo Verhulst ◽  
Joke Soer ◽  
Kees Veldman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Confidence Interval ◽  
Sensitivity And Specificity ◽  
False Positive ◽  
E Coli ◽  
Specificity And Sensitivity ◽  
Positive Results

AbstractThe aim of this study was to determine the performance of both cefotaxime and ceftazidime containing agars on the specificity and sensitivity for chromosomal AmpC-hyperproducing and plasmid AmpC harboring Escherichia coli compared to ESBL-producing E. coli and E. coli without ESBL, pAmpC or cAmpC hyperproduction. Second, we evaluated the influence of adding cefoxitin to these agars for detection of both chromosomal AmpC-hyperproducing and plasmid AmpC harboring E. coli.Four different homemade screening agars with cefotaxime (1mg/L), ceftazidime (1mg/L), cefotaxime (1mg/L) with cefoxitin (8mg/L), and ceftazidime (1mg/L) with cefoxitin (8mg/L) were compared to each other for the identification of AmpC producing E. coli. A total of 40 isolates with plasmid encoded AmpC β-lactamases, 40 isolates with alterations in the promoter/attenuator region of the AmpC gene leading to hyperproduction of the β-lactamase, 40 isolates with ESBL genes and 39 isolates lacking both a AmpC and ESBL genotype were used to test the four agars.The sensitivity and specificity were 100% (95% confidence interval (95% CI) 96.1% to 100%) and 48.1% (95% CI 38.6%-60.2%), respectively, for the cefotaxime agar; 100% (95% CI 96.1% to 100%) and 49.41% (95% CI 39.8%-61.4%), respectively, for the ceftazidime agar; 96.3% (95% CI 89.1% to 99.2%) and 77.2% (95% CI 66.7%-85.2%) respectively, for the cefotaxime with cefoxitin agar; 98.8% (95% CI) 92.6% to 99.6%) and 81.0% (95% CI 70.9%-88.3%) respectively, for the ceftazidime agar with cefoxitin. The main reason for false-positive results were ESBL-harboring strains that grew on various agars; therefore, the specificity of each agar reported here was influenced mainly by the proportion of ESBL isolates tested. In conclusion addition of cefoxitin to cefotaxime and ceftazidime containing agars had little influence on sensitivity, but increased specificity for the detection of AmpC in E. coli.

scholarly journals A comparison between minerals-modified glutamate medium and lauryl tryptose lactose broth for the enumeration of Escherichia coli and coliform organisms in water by the multiple tube method

1980 ◽  
Vol 85 (1) ◽  
pp. 35-49 ◽  
Author(s):  
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Water Samples ◽  
Total Coliform ◽  
Faecal Contamination ◽  
E Coli ◽  
Chlorinated Drinking Water ◽  
Laboratory Trial ◽  
Gas Reactions ◽  
Better Than

SummaryIn a multi-laboratory trial, minerals-modified glutamate medium (MMGM) was compared with lauryl tryptose lactose broth (LTLB) in the multiple tube method for the enumeration of coliform organisms, including Escherichia coli, in water. Samples of raw and chlorinated waters yielded a total of 2313 positive tube-reactions with MMGM and 2174 with LTLB. These were interpreted either as E. coli; other coliform organisms; or as false positive reactions. The results at first reading (18 or 24 h) and at 48 h have been analysed statistically in terms of (i) most probable numbers of coliform organisms; (ii) positive reactions and their interpretation; and (iii) whether or not the sample yielded any E. coli or other coliform organisms. All three analyses indicated the same trends. For the detection of E. coli in raw waters LTLB was better than MMGM at 18–24 h, but MMGM was better at 48 h with waters containing small numbers of coliform organisms; for raw waters with greater numbers of organisms, both media performed equally well. Analysis of a subset of samples read at both 18 and 24 h indicated that the superiority of LTLB over MMGM with raw waters disappeared by 24 h. For chlorinated waters, LTLB yielded more positive gas reactions at 18–24 h, but fewer of these were E. coli than with MMGM; at 48 h MMGM was clearly better than LTLB for total coliform organisms including E. coli – especially if the numbers were small. MMGM therefore remains the medium of choice for the detection of E. coli as an indicator of faecal contamination of chlorinated drinking water supplies. It is also better for the detection of small numbers of E. coli in other waters.

scholarly journals Single tube confirmatory tests forEscherichia coli

1980 ◽  
Vol 85 (1) ◽  
pp. 51-57 ◽  
Author(s):  
Keyword(s):  
Escherichia Coli ◽  
False Positive ◽  
False Negative ◽  
Gas Production ◽  
Single Tube ◽  
E Coli ◽  
Peptone Water ◽  
Confirmatory Tests ◽  
Laboratory Trial ◽  
The Difference

SummaryIn a multi-laboratory trial, lauryl tryptose mannitol broth (LTMB) and minerals-modified glutamate medium with added tryptophan (MMGM + T) were compared as single tube tests for the confirmation ofEscherichia coliin water; the confirmed results were also compared with the production of gas from minerals-modified glutamate medium without added tryptophan (MMGM). LTMB and MMGM + T gave similar gas and indole results with about 90% of the water samples in most of the laboratories. When compared with the ‘correct’ results as judged by acid and gas production from lactose peptone water and indole from tryptone water, the difference in the rate of false positive reactions between LTMB and MMGM + T was insignificant; but LTMB gave a significantly lower rate of false negative reactions than MMGM + T. Gas production from MMGM without added tryptophan gave significantly higher rates of both false positive and false negative reactions. Lauryl sulphate is therefore a suitable inhibitory surfactant for use in single tubemedia for the confirmation ofE. coli, which can be recommended.

scholarly journals Microbiological contamination of water pans in Baringo County

2017 ◽  
Vol 7 (3) ◽  
pp. 485-494
Author(s):  
Edith J. Kurui ◽  
George M. Ogendi ◽  
Wilkister N. Moturi
Keyword(s):  
Water Samples ◽  
Membrane Filtration ◽  
Microbial Contamination ◽  
Total Coliforms ◽  
E Coli ◽  
Quality Guidelines ◽  
Significant Difference ◽  
Significant Spatial Variation ◽  
Dry Seasons ◽  
Filtration Technique

Water pans constitute the main source of rural water supply in Baringo County. This study sought to assess the spatial-temporal variation of total coliforms, Escherichia coli, Fecal streptococcus and Salmonella species in the water pans. A sanitary survey was conducted to observe the potential sources of microbial contamination on the water pans. Water was sampled from one protected and five unprotected water pans (n = 6) in the study area for a period of 4 months (June–October 2015). A total of 72 water samples were sampled in triplicate from the water pans for microbial analyses, membrane filtration technique was used in assaying for microbial counts of total coliforms, E. coli, F. streptococcus and Salmonella species in water samples. The results show that there was a significant spatial variation in F. streptococcus amongst the protected and the unprotected water pan sampled sites (p = 0.008), and there was a statistically temporal significant difference (p = 0.001) for total coliforms and Salmonella species during the dry seasons, respectively. Given the prevalence of the selected diseases causing pathogens in water above the WHO drinking water quality guidelines, households are advised to treat the water before use.

Characterization of O157:H7 and Other Escherichia coli Isolates Recovered from Cattle Hides, Feces, and Carcasses†

2004 ◽  
Vol 67 (5) ◽  
pp. 993-998 ◽  
Author(s):  
GENEVIEVE A. BARKOCY-GALLAGHER ◽  
TERRANCE M. ARTHUR ◽  
MILDRED RIVERA-BETANCOURT ◽  
XIANGWU NOU ◽  
STEVEN D. SHACKELFORD ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Seasonal Variations ◽  
False Positive ◽  
Significant Trend ◽  
Escherichia Coli O157 ◽  
Seasonal Prevalence ◽  
E Coli ◽  
Beef Processing ◽  
Positive Results

In a previous study, the seasonal prevalence was reported for stx+ Escherichia coli O157:H7 in feces and on hides and carcasses of cattle at processing. Overall, 1,697 O157:H7 isolates have now been characterized for the incidence of (i) eaeO157, hlyA, stx1, and stx2 in the recovered isolates and (ii) presumptive rough and presumptive nonmotile isolates. Seven O157:H7 isolates (0.4%) lacked stx genes, although they carried eae and hlyA. All but one of the isolates carried both eae and hlyA. Approximately two-thirds of the isolates (64% when one isolate per sample was considered) carried both stx1 and stx2. E. coli O157:H7 cells that harbored both stx1 and stx2 were more often recovered from hides in the fall (79% of the fall hide isolates) and winter (84% of the winter hide isolates) than in the spring (53%) and summer (59%). Isolates recovered from preevisceration carcasses showed a similar but not statistically significant trend. Twenty-three of the 25 O157:H7 isolates carrying stx1 but not stx2 were recovered during summer. Fifteen presumptive rough and 117 presumptive nonmotile stx+ O157:H7 isolates were recovered. Ten (67%) of the presumptive rough isolates were recovered during summer. Ninety-five of the presumptive nonmotile isolates (81%) were recovered during fall. Forty-eight percent of the false-positive isolates (175 of 363) tentatively identified as O157:H7 were O157+ H7− and lacked eaeO157, hlyA, and stx. These data suggest that in beef processing samples (i) there are minor seasonal variations in the prevalence of stx genes among E. coli O157:H7 isolates, (ii) presumptive rough and presumptive nonmotile stx+ O157:H7 isolates are present, (iii) E. coli O157:H7 isolates lacking stx genes may be rare, and (iv) O157+ H7− isolates lacking stx genes can result in many false-positive results.

Tracking an Escherichia coli O157:H7–Contaminated Batch of Leafy Greens through a Pilot-Scale Fresh-Cut Processing Line

2014 ◽  
Vol 77 (9) ◽  
pp. 1487-1494 ◽  
Author(s):  
ANNEMARIE L. BUCHHOLZ ◽  
GORDON R. DAVIDSON ◽  
BRADLEY P. MARKS ◽  
EWEN C. D. TODD ◽  
ELLIOT T. RYSER
Keyword(s):  
Escherichia Coli ◽  
Membrane Filtration ◽  
Fluorescent Protein ◽  
Pilot Scale ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Leafy Greens ◽  
Iceberg Lettuce ◽  
Processing Line ◽  
Fresh Cut

Cross-contamination of fresh-cut leafy greens with residual Escherichia coli O157:H7–contaminated product during commercial processing was likely a contributing factor in several recent multistate outbreaks. Consequently, radicchio was used as a visual marker to track the spread of the contaminated product to iceberg lettuce in a pilot-scale processing line that included a commercial shredder, step conveyor, flume tank, shaker table, and centrifugal dryer. Uninoculated iceberg lettuce (45 kg) was processed, followed by 9.1 kg of radicchio (dip inoculated to contain a four-strain, green fluorescent protein–labeled nontoxigenic E. coli O157:H7 cocktail at 106 CFU/g) and 907 kg (2,000 lb) of uninoculated iceberg lettuce. After collecting the lettuce and radicchio in about 40 bags (~22.7 kg per bag) along with water and equipment surface samples, all visible shreds of radicchio were retrieved from the bags of shredded product, the equipment, and the floor. E. coli O157:H7 populations were quantified in the lettuce, water, and equipment samples by direct plating with or without prior membrane filtration on Trypticase soy agar containing 0.6% yeast extract and 100 ppm of ampicillin. Based on triplicate experiments, the weight of radicchio in the shredded lettuce averaged 614.9 g (93.6%), 6.9 g (1.3%), 5.0 g (0.8%), and 2.8 g (0.5%) for bags 1 to 10, 11 to 20, 21 to 30, and 31 to 40, respectively, with mean E. coli O157:H7 populations of 1.7, 1.2, 1.1, and 1.1 log CFU/g in radicchio-free lettuce. After processing, more radicchio remained on the conveyor (9.8 g; P < 0.05), compared with the shredder (8.3 g), flume tank (3.5 g), and shaker table (0.1 g), with similar E. coli O157:H7 populations (P > 0.05) recovered from all equipment surfaces after processing. These findings clearly demonstrate both the potential for the continuous spread of contaminated lettuce to multiple batches of product during processing and the need for improved equipment designs that minimize the buildup of residual product during processing.

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.

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