scholarly journals Enumeration of Escherichia coli and determination of Salmonella spp. and verotoxigenic Escherichia coli in shellfish (Mytilus galloprovincialis and Ruditapes decussatus) harvested in Sardinia, Italy

2020 ◽  
Vol 9 (4) ◽  
Author(s):  
Sonia Lamon ◽  
Francesca Piras ◽  
Domenico Meloni ◽  
Vanessa Agus ◽  
Gabriella Porcheddu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mytilus Galloprovincialis ◽  
Reference Method ◽  
Most Probable Number ◽  
Ruditapes Decussatus ◽  
The European Union ◽  
Salmonella Spp ◽  
Probable Number ◽  
Harvesting Time ◽  
E Coli

The aim of the present study was to evaluate the occurrence of Salmonella spp., Verotoxigenic E. Coli (VTEC) and enumerate E. coli in shellfish (Mytilus galloprovincialis and Ruditapes decussatus) collected before and after depuration from two class B harvesting areas located in Sardinia (Italy). All the samples were analyzed for Salmonella spp. detection according to European Commission Regulation (EC) 2073/2005 and examined using the five tube Most-Probable-Number (MPN) method for enumeration of E. coli in accordance with the European Union reference method ISO 16649-3:2015. E. coli VTEC was investigated following a direct multiplex Polymerase Chain Reaction (PCR) screening test. The enumeration of E. coli met the European law limit for Class A areas of 230 MPN/100g. The averaged enumeration of E. coli in samples of M. galloprovincialis and R. decussatus collected at the harvesting time was 39 and 37 MPN/100 g respectively. The average contamination levels in samples collected after purification were 58 MPN/100g (M. galloprovincialis) and 32 MPN/100 g (R. decussatus). E. coli VTEC was not detected, on the contrary, Salmonella ser. Typhimurium was detected in one sample of M. galloprovincialis and in one sample of R. decussatus collected at the harvesting time. No significant associations were observed between E. coli levels in shellfish and environmental parameters of water or with the detection of Salmonella ser. Typhimurium in M. galloprovincialis and R. decussatus samples. Nevertheless, the occurrence of Salmonella ser. Typhimurium, involved in human infection outbreaks, should be considered a potential risk for consumers.

scholarly journals Sources of variation of Escherichia coli concentrations in bivalve molluscs

2012 ◽  
Vol 11 (1) ◽  
pp. 78-83 ◽  
Author(s):  
R. J. Lee ◽  
R. Silk
Keyword(s):  
Escherichia Coli ◽  
Feeding Activity ◽  
Most Probable Number ◽  
Sampling Occasion ◽  
The European Union ◽  
Bivalve Molluscs ◽  
Normal Filter ◽  
Probable Number ◽  
E Coli ◽  
Production Areas

Bivalve molluscs can concentrate contaminants, including pathogenic microorganisms, from the water column during their normal filter-feeding activity. In the European Union, the risk of human and animal faecal contamination in bivalves is estimated by determining the concentration of Escherichia coli in time-series samples from production areas. A structured field study was undertaken to determine the extent to which such concentrations varied between sites, sampling occasions and shellfish species and to determine the residual variability of the method. E. coli was enumerated in three species of bivalve mollusc (Crassostrea gigas, Mytilus spp. and Pecten maximus) co-located in each of three geographically separate commercial shellfisheries. The data were subjected to analysis of variance (ANOVA). This showed that the effects of site, sampling occasion, species and site/sampling occasion interaction were all significant. The proportion of variation due to site was markedly greater than that due to other factors. Post-ANOVA analysis showed that the concentration of E. coli in P. maximus was significantly higher than in the other two species. Mytilus spp. and C. gigas exhibited comparable levels of E. coli. The observed standard deviation of the most probable number method in the study was 0.33 log10.

scholarly journals Two Rapid Methods for Detection of Escherichia coli Exceeding 104/g Action Levels: Precollaborative Study

2006 ◽  
Vol 89 (5) ◽  
pp. 1317-1326
Author(s):  
Michael A Grant ◽  
Jane S Wernberg ◽  
Khanh T Van ◽  
Angelina M Albert
Keyword(s):  
Escherichia Coli ◽  
Membrane Filtration ◽  
Reference Method ◽  
Most Probable Number ◽  
Lauryl Sulfate ◽  
Probable Number ◽  
E Coli ◽  
Potential Alternative ◽  
Aoac Method ◽  
Action Levels

Abstract The current AOAC Method 966.24 for enumeration of Escherichia coli in foods uses a most probable number (MPN) procedure with extensive confirmation steps. Two new methods based on membrane filtration (MF) were compared to the MPN reference method for detection of high levels of E. coli in 5 food types, some of which represent categories for which the U.S. Food and Drug Administration (FDA) mandates additional testing if an action level of 104/g E. coli is exceeded. Ground beef, which is not FDA regulated, was also tested. The 5 food types were all inoculated at 3 levels: 102/g, ≥104/g, and ≥105/g E. coli. An MF protocol using either m-ColiBlue24® (CB) or lauryl sulfate tryptose plus BCIG (LST/BCIG) was an effective potential alternative to the reference method. Sensitivity and specificity for both CB and LST/BCIG were 98 and 100%, respectively. Agreement between MPN and both CB and LST/BCIG was 98%. The 2 proposed methods allow completion of both presumptive and confirmatory steps in 13 days, whereas the reference method requires as many as 11 days. Exclusivity testing with 50 non-E. coli strains indicated 100% were correctly ruled out by the proposed protocols. Inclusivity testing was used to determine whether typical results were obtained after incubation of E. coli cultures on CB or LST/BCIG for 24 h. Of 50 E. coli strains tested, 100% yielded typical results after incubation on CB, and 98% yielded typical results after incubation on LST/BCIG.

scholarly journals Comparison of Colilert-18 with miniaturised most probable number method for monitoring of Escherichia coli in bathing water

2015 ◽  
Vol 14 (1) ◽  
pp. 121-131 ◽  
Author(s):  
Ananda Tiwari ◽  
Seppo I. Niemelä ◽  
Asko Vepsäläinen ◽  
Jarkko Rapala ◽  
Seija Kalso ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
False Negative ◽  
Reference Method ◽  
Water Quality Monitoring ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Probable Number ◽  
E Coli ◽  
Negative Results ◽  
Bathing Water

The purpose of this equivalence study was to compare an alternative method, Colilert-18 Quanti-Tray (ISO 9308-2) with the European bathing water directive (2006/7/EC) reference method, the miniaturised most probable number (MMPN) method (ISO 9308-3), for the analysis of Escherichia coli. Six laboratories analysed a total of 263 bathing water samples in Finland. The comparison was carried out according to ISO 17994:2004. The recovery of E. coli using the Colilert-18 method was 7.0% and 8.6% lower than that of the MMPN method after 48 hours and 72 hours of incubation, respectively. The confirmation rate of presumptive E. coli-positive wells in the Colilert-18 and MMPN methods was high (97.8% and 98.0%, respectively). However, the testing of presumptive E. coli-negative but coliform bacteria-positive (yellow but not fluorescent) Colilert-18 wells revealed 7.3% false negative results. There were more false negatives in the naturally contaminated waters than in the samples spiked with waste water. The difference between the recovery of Colilert-18 and the MMPN method was considered not significant, and subsequently the methods are considered as equivalent for bathing water quality monitoring in Finland. Future bathing water method equivalence verification studies may use the data reported herein. The laboratories should make sure that any wells showing even minor fluorescence will be determined as positive for E. coli.

scholarly journals Lessons from a large outbreak of Escherichia coli O157[ratio ]H7 infections: insights into the infectious dose and method of widespread contamination of hamburger patties

1999 ◽  
Vol 122 (2) ◽  
pp. 185-192 ◽  
Author(s):  
J. TUTTLE ◽  
T. GOMEZ ◽  
M. P. DOYLE ◽  
J. G. WELLS ◽  
T. ZHAO ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ground Beef ◽  
Most Probable Number ◽  
Strong Argument ◽  
Probable Number ◽  
Infectious Dose ◽  
E Coli ◽  
Beef Patties ◽  
Large Outbreak ◽  
Microbiological Testing

Between November 1992 and February 1993, a large outbreak of Escherichia coli O157[ratio ]H7 infections occurred in the western USA and was associated with eating ground beef patties at restaurants of one fast-food chain. Restaurants that were epidemiologically linked with cases served patties produced on two consecutive dates; cultures of recalled ground beef patties produced on those dates yielded E. coli O157[ratio ]H7 strains indistinguishable from those isolated from patients, confirming the vehicle of illness. Seventy-six ground beef patty samples were cultured quantitatively for E. coli O157[ratio ]H7. The median most probable number of organisms was 1·5 per gram (range, <0·3–15) or 67·5 organisms per patty (range, <13·5–675). Correlation of the presence of E. coli O157[ratio ]H7 with other bacterial indicators yielded a significant association between coliform count and the presence of E. coli O157[ratio ]H7 (P=0·04). A meat traceback to investigate possible sources of contamination revealed cattle were probably initially colonized with E. coli O157[ratio ]H7, and that their slaughter caused surface contamination of meat, which once combined with meat from other sources, resulted in a large number of contaminated ground beef patties. Microbiological testing of meat from lots consumed by persons who became ill was suggestive of an infectious dose for E. coli O157[ratio ]H7 of fewer than 700 organisms. These findings present a strong argument for enforcing zero tolerance for this organism in processed food and for markedly decreasing contamination of raw ground beef. Process controls that incorporate microbiological testing of meat may assist these efforts.

scholarly journals Mechanically Ventilated Broiler Sheds: a Possible Source of Aerosolized Salmonella, Campylobacter, and Escherichia coli

2009 ◽  
Vol 75 (23) ◽  
pp. 7417-7425 ◽  
Author(s):  
H. N. Chinivasagam ◽  
T. Tran ◽  
L. Maddock ◽  
A. Gale ◽  
P. J. Blackall
Keyword(s):  
Escherichia Coli ◽  
Most Probable Number ◽  
Production Cycle ◽  
Airborne Bacteria ◽  
Probable Number ◽  
Mechanically Ventilated ◽  
The Public ◽  
E Coli ◽  
Poultry Farms ◽  
Low Levels

ABSTRACT This study assessed the levels of two key pathogens, Salmonella and Campylobacter, along with the indicator organism Escherichia coli in aerosols within and outside poultry sheds. The study ranged over a 3-year period on four poultry farms and consisted of six trials across the boiler production cycle of around 55 days. Weekly testing of litter and aerosols was carried out through the cycle. A key point that emerged is that the levels of airborne bacteria are linked to the levels of these bacteria in litter. This hypothesis was demonstrated by E. coli. The typical levels of E. coli in litter were ∼108 CFU g−1 and, as a consequence, were in the range of 102 to 104 CFU m−3 in aerosols, both inside and outside the shed. The external levels were always lower than the internal levels. Salmonella was only present intermittently in litter and at lower levels (103 to 105 most probable number [MPN] g−1) and consequently present only intermittently and at low levels in air inside (range of 0.65 to 4.4 MPN m−3) and once outside (2.3 MPN m−3). The Salmonella serovars isolated in litter were generally also isolated from aerosols and dust, with the Salmonella serovars Chester and Sofia being the dominant serovars across these interfaces. Campylobacter was detected late in the production cycle, in litter at levels of around 107 MPN g−1. Campylobacter was detected only once inside the shed and then at low levels of 2.2 MPN m−3. Thus, the public health risk from these organisms in poultry environments via the aerosol pathway is minimal.

scholarly journals GREEN SYNTHESIS OF MAGNETIC IRON NANOPARTICLES USING MEDICINAL PLANT TRIDAX PROCUMBENS LEAF EXTRACTS AND ITS APPLICATION AS AN ANTIMICROBIAL AGENT AGAINST E. COLI

2020 ◽  
pp. 34-39
Author(s):  
YOJANA Y. PATIL ◽  
VAISHNVI B. SUTAR ◽  
ARPITA P. TIWARI
Keyword(s):  
Escherichia Coli ◽  
Magnetic Nanoparticles ◽  
Iron Nanoparticles ◽  
Most Probable Number ◽  
Leaf Extracts ◽  
Probable Number ◽  
Gram Negative ◽  
E Coli ◽  
Tridax Procumbens ◽  
Escherichia Coli Bacteria

Objective: The present study was aimed at the biological synthesis of magnetic iron nanoparticles by using the plant extract of Tridax procumbens and also to study their antimicrobial property against gram-negative bacteria (Escherichia coli). Methods: The synthesis of magnetic iron nanoparticles was carried out by the co-precipitation method using biological methods like plant extract as reducing agent and capping agents are biocompatible and non-hazardous. These nanoparticles were characterized by UV-Visible spectroscopy, XRD (X-Ray Diffraction), and SEM (Scanning Electron Microscope). As well as antibacterial activity of the nanoparticles was carried out by agar well diffusion method and Most Probable Number (MPN) method against gram-negative E. coli (Escherichia coli) bacteria. Results: The average crystallite size of Magnetic Nanoparticles (MNPs) was found to be 72 nm by X-ray diffraction. The optical absorption band at wavelengths of 240 nm and 402 nm was obtained from the UV Visible spectrum. Spherical shape morphology was observed in SEM studies. The antibacterial assay clearly expressed that E. coli showed a maximum zone of inhibition (15±0.15 mm) at 2 mg/ml and 1 mg/ml concentration was found for Magnetic Nanoparticles. In the Most Probable Number (MPN) test it is seen that the bacterial count is reduced after adding synthesized NPs into the water sample. Conclusion: The results of the present study conclude that the Magnetic Nanoparticles synthesized using Tridax procumbens leaf extracts is found to be stable and show good antibacterial activity against gram-negative (Escherichia coli) bacteria.

scholarly journals Escherichia coli O157:H7 in Drin Prevalence of Escherichia coli O157:H7 in Drink from Different Food Premises in Kota Samarahan Sarawak

2019 ◽  
Vol 2 (2) ◽  
pp. a13-19
Author(s):  
ELEXSON NILLIAN ◽  
AMIZA NUR ◽  
DIYANA NUR ◽  
AMIRAH ZAKIRAH ◽  
GRACE BEBEY
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Plain Water ◽  
Escherichia Coli O157 ◽  
Probable Number ◽  
E Coli ◽  
Preventive Actions ◽  
Stx1 Gene

Contamination of drinks with E. coli O157:H7 served in food premises such as restaurants can cause haemorrhagic colitis and haemolytic uremic syndrome to humans. The presence or absence of faecal pathogen was demonstrated using coliform group as indicator microorganisms. Therefore, this study was conducted to detect the presence of E. coli O157:H7 in drinking water from food restaurant premise in Kota Samarahan and Kuching to ensure safe and potable drinking water is served to the consumer. A total of thirty (n=30) drink samples including six types of each of the samples are cold plain water, iced tea, iced milo, syrup and iced milk tea. Most Probable Number (MPN) procedure was used in this study to enumerate the MPN values of coliform bacteria in each drink collected. A total of 53.33% (16/30) of the drink samples showed positive E. coli detection. Then, the PCR assay showed 6.25% (one out of 16 isolates) samples were positive and carried stx1 gene produced by E. coli O157:H7 in iced milo sample types. This study showed the drinks collected from food premises was contaminated with faecal contamination, which was not safe to drink by the consumer. Therefore, preventive actions should be taken to prevent foodborne illness outbreak in future

Survival of Escherichia coli on Lettuce under Field Conditions Encountered in the Northeastern United States

2017 ◽  
Vol 80 (7) ◽  
pp. 1214-1221 ◽  
Author(s):  
Daniel L. Weller ◽  
Jasna Kovac ◽  
Sherry Roof ◽  
David J. Kent ◽  
Jeffrey I. Tokman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Microbial Contamination ◽  
Field Conditions ◽  
Risk Assessments ◽  
Most Probable Number ◽  
Northeastern United States ◽  
Probable Number ◽  
E Coli ◽  
The Relationship ◽  
Sample Time

ABSTRACT Although wildlife intrusion and untreated manure have been associated with microbial contamination of produce, relatively few studies have examined the survival of Escherichia coli on produce under field conditions following contamination (e.g., via splash from wildlife feces). This experimental study was performed to estimate the die-off rate of E. coli on preharvest lettuce following contamination with a fecal slurry. During August 2015, field-grown lettuce was inoculated via pipette with a fecal slurry that was spiked with a three-strain cocktail of rifampin-resistant nonpathogenic E. coli. Ten lettuce heads were harvested at each of 13 time points following inoculation (0, 2.5, 5, and 24 h after inoculation and every 24 h thereafter until day 10). The most probable number (MPN) of E. coli on each lettuce head was determined, and die-off rates were estimated. The relationship between sample time and the log MPN of E. coli per head was modeled using a segmented linear model. This model had a breakpoint at 106 h (95% confidence interval = 69, 142 h) after inoculation, with a daily decrease of 0.70 and 0.19 log MPN for 0 to 106 h and 106 to 240 h following inoculation, respectively. These findings are consistent with die-off rates obtained in similar studies that assessed E. coli survival on produce following irrigation. Overall, these findings provide die-off rates for E. coli on lettuce that can be used in future quantitative risk assessments.

A Bacteriological Survey of Raw Ground Beef1

1977 ◽  
Vol 40 (11) ◽  
pp. 790-794 ◽  
Author(s):  
JAMES F. FOSTER ◽  
JAMES L. FOWLER ◽  
WARREN C. LADIGES
Keyword(s):  
Escherichia Coli ◽  
Microbiological Quality ◽  
Retail Store ◽  
Most Probable Number ◽  
Probable Number ◽  
E Coli ◽  
Fecal Streptococci ◽  
Plate Counts ◽  
Aerobic And Anaerobic

The microbiological quality of 150 units of raw ground beef obtained from a local retail store was determined. The range of aerobic plate counts was from 6.9 × 104 to 8.3 × 107/g. By using the most probable number method 96.7% of the 150 units were positive for coliforms, 94.7% for Escherichia coli and 61.3% for Staphylococcus aureus. By the plate methods, 99.3% of the units were positive for fecal streptococci and 56% were positive for Clostridium perfringens. No salmonellae were isolated. Aerobic and anaerobic organisms were isolated and identified. E. coli was the most frequently isolated aerobe followed by organisms in the Klebsiella-Enterobacter group. Among the anaerobic isolates, C. perfringens was the organism most frequently encountered.

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

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