scholarly journals Detection of Viable but Nonculturable Escherichia coli O157:H7 Bacteria in Drinking Water and River Water

2008 ◽  
Vol 74 (5) ◽  
pp. 1502-1507 ◽  
Author(s):  
Yanming Liu ◽  
Ainslie Gilchrist ◽  
Jing Zhang ◽  
Xing-Fang Li
Keyword(s):  
Escherichia Coli ◽  
River Water ◽  
Water Samples ◽  
Limit Of Detection ◽  
Tap Water ◽  
Environmental Water ◽  
Bacterial Cells ◽  
Viable But Nonculturable ◽  
E Coli ◽  
Extraction And Purification

ABSTRACT A sensitive method for specific detection of viable Escherichia coli O157:H7 cells, including viable but nonculturable (VBNC) cells, in water samples was developed. This method involved capture of the bacterial cells on a low-protein-binding membrane and direct extraction and purification of RNA followed by reverse transcription-PCR and electronic microarray detection of the rfbE and fliC genes of E. coli O157:H7. It detected as few as 1 CFU of E. coli O157:H7 in diluted cultures, 3 to 4 CFU/liter in tap water, 7 CFU/liter in river water, and 50 VBNC cells in 1 liter of river water, demonstrating the best limit of detection reported to date for VBNC cells in environmental water samples.

scholarly journals Diversity, virulence and antibiogram traits of Escherichia coli recovered from potable water sources in Gharbia, Egypt

2020 ◽  
Vol 18 (3) ◽  
pp. 430-438
Author(s):  
Walid Elmonir ◽  
Etab Mohamed Abo Remela ◽  
Yasmine Alwakil
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Water Samples ◽  
Potable Water ◽  
Multiple Drug Resistance ◽  
Tap Water ◽  
Water Sources ◽  
Multiple Drug ◽  
Well Water ◽  
E Coli

Abstract This study aimed to assess the public health risk of coliforms and Escherichia coli contamination of potable water sources in Egypt. A total of 150 water samples (100 tap and 50 well) were collected from five districts in Gharbia governorate, Egypt. High rates of coliforms contamination were recorded in 52 and 76% of examined tap and well water samples, respectively. E. coli strains were detected in 16% of the water samples (15% tap water and 18% well water; 23.7% rural and 8.1% urban). Rural water sources were 3.5 times more likely to be contaminated than urban sources (P = 0.01). Eight (33.3%) E. coli isolates were Shiga toxin-producing E. coli (STEC). Multiple drug resistance (MDR) was observed for 62.5% of the isolates. Seven (29.2%) E. coli isolates harboured at least one of the extended-spectrum beta-lactamase (ESBL) genes. The majority (87.5%) of the STEC isolates were MDRs and harboured ESBL genes. STEC isolates were significantly more likely to resist six classes of antibiotics than non-STEC isolates. This is the first report of potable water contamination with MDR-STEC in Egypt. This study highlights an alarming public health threat that necessitates preventive interventions for public and environmental safety.

scholarly journals Prevalence of Legionella spp. and Escherichia coli in the drinking water distribution system of Wrocław (Poland)

2020 ◽  
Vol 20 (3) ◽  
pp. 1083-1090
Author(s):  
M. Wolf-Baca ◽  
A. Siedlecka
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Legionella Pneumophila ◽  
Water Samples ◽  
Distribution System ◽  
Water Distribution ◽  
Tap Water ◽  
Potential Threat ◽  
E Coli ◽  
Drinking Water Distribution

Abstract Drinking water should be free from bacterial pathogens that threaten human health. The most recognised waterborne opportunistic pathogens, dwelling in tap water, are Legionella pneumophila and Escherichia coli. Drinking water samples were tested for the presence of Legionella spp., L. pneumophila, and E. coli in overall sample microbiomes using a quantitative real-time polymerase chain reaction (qPCR) approach. The results indicate a rather low contribution of Legionella spp. in total bacteria in the tested samples, but L. pneumophila was not detected in any sample. E. coli was detected in only one sample, but at a very low level. The qacEΔ1 gene, conferring resistance to quaternary ammonium compounds, was also not detected in any sample. The results point to generally sufficient quality of drinking water, although the presence of Legionella spp. in tap water samples suggests proliferation of these bacteria in heating units, causing a potential threat to consumer health.

scholarly journals Cell Surface Display of Poliovirus Receptor on Escherichia coli, a Novel Method for Concentrating Viral Particles in Water

2011 ◽  
Vol 77 (15) ◽  
pp. 5141-5148 ◽  
Author(s):  
Morteza Abbaszadegan ◽  
Absar Alum ◽  
Hamed Abbaszadegan ◽  
Valerie Stout
Keyword(s):  
Escherichia Coli ◽  
Cell Surface ◽  
Water Samples ◽  
Stop Codon ◽  
Surface Display ◽  
Cell Surface Display ◽  
Bacterial Cells ◽  
Content Type ◽  
E Coli ◽  
Poliovirus Receptor

ABSTRACTThe lack of efficient methods for concentrating viruses in water samples leads to underreporting of viral contamination in source water. A novel strategy for viral concentration was developed using the expression of target virus receptors on bacterial cells. Poliovirus type 1, the most studied enterovirus, was used as a surrogate for enteric viruses. The human poliovirus receptor (hPVR) gene was expressed on the surface ofEscherichia colicells by using the ice nucleation protein (INP) gene. ThehPVRgene was ligated to the 3′ end of theINPgene after the removal of the stop codon. The resulting open reading frame (ORF) was used for the projection of hPVR onto the outer membrane ofE. coli. Gene expression was tested by SDS-PAGE, Western blot, and dot blot analyses, and virion capture ability was confirmed by transmission electron microscopy. The application of engineeredE. colicells for capturing viruses in 1-liter samples of source and drinking water resulted in 75 to 99% procedural recovery efficiency. Cell surface display of viral receptors on bacterial cells opens a new prospect for an efficient and inexpensive alternative tool for capturing and concentrating waterborne viruses in water samples.

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.

scholarly journals Escherichia coli Contamination of Vegetables Grown in Soils Fertilized with Noncomposted Bovine Manure: Garden-Scale Studies

2004 ◽  
Vol 70 (11) ◽  
pp. 6420-6427 ◽  
Author(s):  
Steven C. Ingham ◽  
Jill A. Losinski ◽  
Matthew P. Andrews ◽  
Jane E. Breuer ◽  
Jeffry R. Breuer ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Tap Water ◽  
Silty Clay ◽  
Silt Loam ◽  
Silty Clay Loam ◽  
Manure Application ◽  
E Coli ◽  
Fertilized Soil ◽  
Low Levels ◽  
National Organic Program

ABSTRACT In this study we tested the validity of the National Organic Program (NOP) requirement for a ≥120-day interval between application of noncomposted manure and harvesting of vegetables grown in manure-fertilized soil. Noncomposted bovine manure was applied to 9.3-m2 plots at three Wisconsin sites (loamy sand, silt loam, and silty clay loam) prior to spring and summer planting of carrots, radishes, and lettuce. Soil and washed (30 s under running tap water) vegetables were analyzed for indigenous Escherichia coli. Within 90 days, the level of E. coli in manure-fertilized soil generally decreased by about 3 log CFU/g from initial levels of 4.2 to 4.4 log CFU/g. Low levels of E. coli generally persisted in manure-fertilized soil for more than 100 days and were detected in enriched soil from all three sites 132 to 168 days after manure application. For carrots and lettuce, at least one enrichment-negative sample was obtained ≤100 days after manure application for 63 and 88% of the treatments, respectively. The current ≥120-day limit provided an even greater likelihood of not detecting E. coli on carrots (≥1 enrichment-negative result for 100% of the treatments). The rapid maturation of radishes prevented conclusive evaluation of a 100- or 120-day application-to-harvest interval. The absolute absence of E. coli from vegetables harvested from manure-fertilized Wisconsin soils may not be ensured solely by adherence to the NOP ≥120-day limit. Unless pathogens are far better at colonizing vegetables than indigenous E. coli strains are, it appears that the risk of contamination for vegetables grown in Wisconsin soils would be elevated only slightly by reducing the NOP requirement to ≥100 days.

One-Step Preparation of Competent E. coli: Transformation and Storage of Bacterial Cells in the Same Solution

2021 ◽  
Vol 2021 (11) ◽  
pp. pdb.prot101212 ◽  
Author(s):  
Michael R. Green ◽  
Joseph Sambrook
Keyword(s):  
Escherichia Coli ◽  
Convenient Method ◽  
Plasmid Dna ◽  
Transformation Efficiency ◽  
Bacterial Cells ◽  
E Coli ◽  
One Step ◽  
And Storage

This protocol describes a convenient method for the preparation, use, and storage of competent Escherichia coli. The reported transformation efficiency of this method is ∼5 × 107 transformants/µg of plasmid DNA.

PBP4 and PBP5 are involved in regulating exopolysaccharide synthesis during Escherichia coli biofilm formation

Microbiology ◽  
2021 ◽  
Vol 167 (3) ◽  
Author(s):  
Sathi Mallick ◽  
Shanti Kiran ◽  
Tapas Kumar Maiti ◽  
Anindya S. Ghosh
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Type Species ◽  
Pentose Phosphate ◽  
Bacterial Cells ◽  
Surface Adhesion ◽  
Content Type ◽  
Penicillin Binding Proteins ◽  
E Coli ◽  
Link Type

Escherichia coli low-molecular-mass (LMM) Penicillin-binding proteins (PBPs) help in hydrolysing the peptidoglycan fragments from their cell wall and recycling them back into the growing peptidoglycan matrix, in addition to their reported involvement in biofilm formation. Biofilms are external slime layers of extra-polymeric substances that sessile bacterial cells secrete to form a habitable niche for themselves. Here, we hypothesize the involvement of Escherichia coli LMM PBPs in regulating the nature of exopolysaccharides (EPS) prevailing in its extra-polymeric substances during biofilm formation. Therefore, this study includes the assessment of physiological characteristics of E. coli CS109 LMM PBP deletion mutants to address biofilm formation abilities, viability and surface adhesion. Finally, EPS from parent CS109 and its ΔPBP4 and ΔPBP5 mutants were purified and analysed for sugars present. Deletions of LMM PBP reduced biofilm formation, bacterial adhesion and their viability in biofilms. Deletions also diminished EPS production by ΔPBP4 and ΔPBP5 mutants, purification of which suggested an increased overall negative charge compared with their parent. Also, EPS analyses from both mutants revealed the appearance of an unusual sugar, xylose, that was absent in CS109. Accordingly, the reason for reduced biofilm formation in LMM PBP mutants may be speculated as the subsequent production of xylitol and a hindrance in the standard flow of the pentose phosphate pathway.

Changes in Aerobic Plate and Escherichia coli–Coliform Counts and in Populations of Inoculated Foodborne Pathogens on Inshell Walnuts during Storage

2016 ◽  
Vol 79 (7) ◽  
pp. 1143-1153 ◽  
Author(s):  
JOHN C. FRELKA ◽  
GORDON R. DAVIDSON ◽  
LINDA J. HARRIS
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Relative Humidity ◽  
Low Temperature ◽  
Foodborne Pathogens ◽  
Limit Of Detection ◽  
Sampling Time ◽  
E Coli ◽  
Plate Counts ◽  
Forced Air

ABSTRACT After harvest, inshell walnuts are dried using low-temperature forced air and are then stored in bins or silos for up to 1 year. To better understand the survival of bacteria on inshell walnuts, aerobic plate counts (APCs) and Escherichia coli–coliform counts (ECCs) were evaluated during commercial storage (10 to 12°C and 63 to 65% relative humidity) over 9 months. APCs decreased by 1.4 to 2.0 log CFU per nut during the first 5 months of storage, and ECCs decreased by 1.3 to 2.2 log CFU per nut in the first month of storage. Through the remaining 4 to 8 months of storage, APCs and ECCs remained unchanged (P > 0.05) or decreased by <0.15 log CFU per nut per month. Similar trends were observed on kernels extracted from the inshell walnuts. APCs and ECCs were consistently and often significantly higher on kernels extracted from visibly broken inshell walnuts than on kernels extracted from visibly intact inshell walnuts. Parameters measured in this study were used to determine the survival of five-strain cocktails of E. coli O157:H7, Listeria monocytogenes, and Salmonella inoculated onto freshly hulled inshell walnuts (~8 log CFU/g) after simulated commercial drying (10 to 12 h; 40°C) and simulated commercial storage (12 months at 10°C and 65% relative humidity). Populations declined by 2.86, 5.01, and 4.40 log CFU per nut for E. coli O157:H7, L. monocytogenes, and Salmonella, respectively, after drying and during the first 8 days of storage. Salmonella populations changed at a rate of −0.33 log CFU per nut per month between days 8 and 360, to final levels of 2.83 ± 0.79 log CFU per nut. E. coli and L. monocytogenes populations changed by −0.17 log CFU per nut per month and −0.26 log CFU per nut per month between days 8 and 360, respectively. For some samples, E. coli or L. monocytogenes populations were below the limit of detection by plating (0.60 log CFU per nut) by day 183 or 148, respectively; at least one of the six samples was positive at each subsequent sampling time by either plating or by enrichment.

A Comparison of Hand Washing Techniques To Remove Escherichia coli and Caliciviruses under Natural or Artificial Fingernails

2003 ◽  
Vol 66 (12) ◽  
pp. 2296-2301 ◽  
Author(s):  
CHIA-MIN LIN ◽  
FONE-MAO WU ◽  
HOI-KYUNG KIM ◽  
MICHAEL P. DOYLE ◽  
BARRY S. MICHAELS ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Tap Water ◽  
Hand Washing ◽  
Microbial Populations ◽  
Liquid Soap ◽  
E Coli ◽  
Hand Sanitizer ◽  
Before And After ◽  
Escherichia Coli Jm109 ◽  
Viral Indicators

Compared with other parts of the hand, the area beneath fingernails harbors the most microorganisms and is most difficult to clean. Artificial fingernails, which are usually long and polished, reportedly harbor higher microbial populations than natural nails. Hence, the efficacy of different hand washing methods for removing microbes from natural and artificial fingernails was evaluated. Strains of nonpathogenic Escherichia coli JM109 and feline calicivirus (FCV) strain F9 were used as bacterial and viral indicators, respectively. Volunteers with artificial or natural nails were artificially contaminated with ground beef containing E. coli JM109 or artificial feces containing FCV. Volunteers washed their hands with tap water, regular liquid soap, antibacterial liquid soap, alcohol-based hand sanitizer gel, regular liquid soap followed by alcohol gel, or regular liquid soap plus a nailbrush. The greatest reduction of inoculated microbial populations was obtained by washing with liquid soap plus a nailbrush, and the least reduction was obtained by rubbing hands with alcohol gel. Lower but not significantly different (P > 0.05) reductions of E. coli and FCV counts were obtained from beneath artificial than from natural fingernails. However, significantly (P ≤ 0.05) higher E. coli and FCV counts were recovered from hands with artificial nails than from natural nails before and after hand washing. In addition, microbial cell numbers were correlated with fingernail length, with greater numbers beneath fingernails with longer nails. These results indicate that best practices for fingernail sanitation of food handlers are to maintain short fingernails and scrub fingernails with soap and a nailbrush when washing hands.

Effects of Water Source, Dilution, Storage, and Bacterial and Fecal Loads on the Efficacy of Electrolyzed Oxidizing Water for the Control of Escherichia coli O157:H7

2004 ◽  
Vol 67 (7) ◽  
pp. 1377-1383 ◽  
Author(s):  
S. M. L. STEVENSON ◽  
S. R. COOK ◽  
S. J. BACH ◽  
T. A. McALLISTER
Keyword(s):  
Escherichia Coli ◽  
Bactericidal Activity ◽  
Storage Temperature ◽  
Uv Light ◽  
Tap Water ◽  
Organic Matter Content ◽  
Water Source ◽  
Deionized Water ◽  
Escherichia Coli O157 ◽  
E Coli

To evaluate the potential of using electrolyzed oxidizing (EO) water for controlling Escherichia coli O157:H7 in water for livestock, the effects of water source, electrolyte concentration, dilution, storage conditions, and bacterial or fecal load on the oxidative reduction potential (ORP) and bactericidal activity of EO water were investigated. Anode and combined (7:3 anode:cathode, vol/vol) EO waters reduced the pH and increased the ORP of deionized water, whereas cathode EO water increased pH and lowered ORP. Minimum concentrations (vol/vol) of anode and combined EO waters required to kill 104 CFU/ml planktonic suspensions of E. coli O157:H7 strain H4420 were 0.5 and 2.0%, respectively. Cathode EO water did not inhibit H4420 at concentrations up to 16% (vol/vol). Higher concentrations of anode or combined EO water were required to elevate the ORP of irrigation or chlorinated tap water compared with that of deionized water. Addition of feces to EO water products (0.5% anode or 2.0% combined, vol/vol) significantly reduced (P < 0.001) their ORP values to <700 mV in all water types. A relationship between ORP and bactericidal activity of EO water was observed. The dilute EO waters retained the capacity to eliminate a 104 CFU/ml inoculation of E. coli O157:H7 H4420 for at least 70 h regardless of exposure to UV light or storage temperature (4 versus 24°C). At 95 h and beyond, UV exposure reduced ORP, significantly more so (P < 0.05) in open than in closed containers. Bactericidal activity of EO products (anode or combined) was lost in samples in which ORP value had fallen to ≤848 mV. When stored in the dark, the diluted EO waters retained an ORP of >848 mV and bactericidal efficacy for at least 125 h; with refrigeration (4°C), these conditions were retained for at least 180 h. Results suggest that EO water may be an effective means by which to control E. coli O157:H7 in livestock water with low organic matter content.

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