Prevalence of
            Escherichia coli
            O157:H7 from Cull Dairy Cows in New York State and Comparison of Culture Methods Used during Preharvest Food Safety Investigations

ABSTRACT A number of protocols for the cultural detection of Escherichia coli O157:H7 in clinical fecal specimens have been proposed. In the present study direct plating of cattle feces was compared to three different broth enrichment protocols, i.e., a protocol with modified E. coli broth with novobiocin, a protocol with Trypticase soy broth with cefixime and vancomycin, and a protocol with Gram-Negative Broth with novobiocin, for their relative abilities to detect E. coli O157:H7 in feces. In all enrichment protocols, dilutions of the enrichment broths onto 150-mm sorbitol-MacConkey agar plates to which cefixime and tellurite were added were used along with reading of agar plates at both 24 and 48 h. Fecal samples came from a preharvest food safety project in which feces from New York cull dairy cattle from a northeastern packing plant along with experimentally inoculated adult dairy cow feces were tested. The performances of the broth enrichments were comparable to each other, but the broth enrichments were superior to direct plating in their ability to detect E. coli O157:H7. Regardless of the culture protocol used, recovery of E. coli O157:H7 is more likely from fresh fecal specimens than from frozen samples. An overall prevalence of E. coli O157:H7 fecal shedding by New York cull dairy cattle of 1.3% was found in specimens just before processing at the packing plant.

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Plant-Pathogenic Oomycetes, Escherichia coli Strains, and Salmonella spp. Frequently Found in Surface Water Used for Irrigation of Fruit and Vegetable Crops in New York State

Applied and Environmental Microbiology ◽

10.1128/aem.01012-14 ◽

2014 ◽

Vol 80 (16) ◽

pp. 4814-4820 ◽

Cited By ~ 24

Author(s):

Lisa A. Jones ◽

Randy W. Worobo ◽

Christine D. Smart

Keyword(s):

Escherichia Coli ◽

Water Quality ◽

New York ◽

Food Safety ◽

Surface Water ◽

New York State ◽

Quality Parameters ◽

Content Type ◽

York State ◽

E Coli

ABSTRACTIn the United States, surface water is commonly used to irrigate a variety of produce crops and can harbor pathogens responsible for food-borne illnesses and plant diseases. Understanding when pathogens infest water sources is valuable information for produce growers to improve the food safety and production of these crops. In this study, prevalence data along with regression tree analyses were used to correlate water quality parameters (pH, temperature, turbidity), irrigation site properties (source, the presence of livestock or fowl nearby), and precipitation data to the presence and concentrations ofEscherichia coli,Salmonellaspp., and hymexazol-insensitive (HIS) oomycetes (PhytophthoraandPythiumspp.) in New York State surface waters. A total of 123 samples from 18 sites across New York State were tested forE. coliandSalmonellaspp., of which 33% and 43% were positive, respectively. Additionally, 210 samples from 38 sites were tested for HIS oomycetes, and 88% were found to be positive, with 10 species ofPhytophthoraand 11 species ofPythiumbeing identified from the samples. Regression analysis found no strong correlations between water quality parameters, site factors, or precipitation to the presence or concentration ofE. coliin irrigation sources. ForSalmonella, precipitation (≤0.64 cm) 3 days before sampling was correlated to both presence and the highest counts. Analyses for oomycetes found creeks to have higher average counts than ponds, and higher turbidity levels were associated with higher oomycete counts. Overall, information gathered from this study can be used to better understand the food safety and plant pathogen risks of using surface water for irrigation.

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ASSOCIATION OF ESCHERICHIA COLI SEROGROUP 0111 WITH TWO HOSPITAL OUTBREAKS OF EPIDEMIC DIARRHEA OF THE NEWBORN INFANT IN NEW YORK STATE DURING 1947

PEDIATRICS ◽

10.1542/peds.12.4.377 ◽

1953 ◽

Vol 12 (4) ◽

pp. 377-383

Author(s):

ERWIN NETER ◽

ROBERT F. KORNS ◽

RAY E. TRUSSELL

Keyword(s):

Escherichia Coli ◽

New York ◽

Newborn Infant ◽

New York State ◽

York State ◽

E Coli

Escherichia coli serogroup 0111 was isolated in 1952 as the predominant coliform organism from 5 fecal specimens obtained from 7 infants who were suffering from epidemic diarrhea of the newborn infant during two serious hospital outbreaks of this disease in 1947. The specimens had been preserved at a temperature of -40° to -70° C. for five years. The association of certain serogroups of E. coli and epidemic diarrhea of the newborn infant is discussed.

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Prevalence and Characterization of Extended-Spectrum β-Lactamase-Producing Antibiotic-Resistant Escherichia coli and Klebsiella pneumoniae in Ready-to-Eat Street Foods

Antibiotics ◽

10.3390/antibiotics10070850 ◽

2021 ◽

Vol 10 (7) ◽

pp. 850

Author(s):

Shobha Giri ◽

Vaishnavi Kudva ◽

Kalidas Shetty ◽

Veena Shetty

Keyword(s):

Escherichia Coli ◽

Food Safety ◽

Klebsiella Pneumoniae ◽

Rural Areas ◽

Significant Risk ◽

Microbiological Quality ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Extended Spectrum

As the global urban populations increase with rapid migration from rural areas, ready-to-eat (RTE) street foods are posing food safety challenges where street foods are prepared with less structured food safety guidelines in small and roadside outlets. The increased presence of extended-spectrum-β-lactamase (ESBL) producing bacteria in street foods is a significant risk for human health because of its epidemiological significance. Escherichia coli and Klebsiella pneumoniae have become important and dangerous foodborne pathogens globally for their relevance to antibiotic resistance. The present study was undertaken to evaluate the potential burden of antibiotic-resistant E. coli and K. pneumoniae contaminating RTE street foods and to assess the microbiological quality of foods in a typical emerging and growing urban suburb of India where RTE street foods are rapidly establishing with public health implications. A total of 100 RTE food samples were collected of which, 22.88% were E. coli and 27.12% K. pneumoniae. The prevalence of ESBL-producing E. coli and K. pneumoniae was 25.42%, isolated mostly from chutneys, salads, paani puri, and chicken. Antimicrobial resistance was observed towards cefepime (72.9%), imipenem (55.9%), cefotaxime (52.5%), and meropenem (16.9%) with 86.44% of the isolates with MAR index above 0.22. Among β-lactamase encoding genes, blaTEM (40.68%) was the most prevalent followed by blaCTX (32.20%) and blaSHV (10.17%). blaNDM gene was detected in 20.34% of the isolates. This study indicated that contaminated RTE street foods present health risks to consumers and there is a high potential of transferring multi-drug-resistant bacteria from foods to humans and from person to person as pathogens or as commensal residents of the human gut leading to challenges for subsequent therapeutic treatments.

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Bacteriophages reduce Escherichia coli O157:H7 levels in experimentally inoculated sheep

Canadian Journal of Animal Science ◽

10.4141/cjas08083 ◽

2009 ◽

Vol 89 (2) ◽

pp. 285-293 ◽

Cited By ~ 15

Author(s):

S J Bach ◽

R P Johnson ◽

K. Stanford ◽

T A McAllister

Keyword(s):

Escherichia Coli ◽

Oral Administration ◽

Coliform Bacteria ◽

Escherichia Coli O157 ◽

Fecal Shedding ◽

Total Coliforms ◽

E Coli ◽

Oral Inoculation ◽

Experimental Treatments ◽

And Control

Bacteriophage biocontrol has potential as a means of mitigating the prevalence of Escherichia coli O157:H7 in ruminants. The efficacy of oral administration of bacteriophages for reducing fecal shedding of E. coli O157:H7 by sheep was evaluated using 20 Canadian Arcott rams (50.0 ± 3.0) housed in four rooms (n = 5) in a contained facility. The rams had ad libitum access to drinking water and a pelleted barley-based total mixed ration, delivered once daily. Experimental treatments consisted of administration of E. coli O157:H7 (O157), E. coli O157:H7+bacteriophages (O157+phage), bacteriophages (phage), and control (CON). Oral inoculation of the rams with 109 CFU of a mixture of four nalidixic acid-resistant strains of E. coli O157:H7 was performed on day 0. A mixture of 1010 PFU of bacteriophages P5, P8 and P11 was administered on days -2, -1, 0, 6 and 7. Fecal samples collected on 14 occasions over 21 d were analyzed for E. coli O157:H7, total E. coli, total coliforms and bacteriophages. Sheep in treatment O157+phage shed fewer (P < 0.05) E. coli O157:H7 than did sheep in treatment O157. Populations of total coliforms and total E. coli were similar (P < 0.05) among treatments, implying that bacteriophage lysis of non-target E. coli and coliform bacteria in the gastrointestinal tract did not occur. Bacteriophage numbers declined rapidly over 21 d, which likely reduced the chance of collision between bacteria and bacteriophage. Oral administration of bacteriophages reduced shedding of E. coli O157:H7 by sheep, but a delivery system that would protect bacteriophages during passage through the intestine may increase the effectiveness of this strategy as well as allow phage to be administered in the feed.Key words: Escherichia coli O157:H7, bacteriophage, sheep, environment, coliforms

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Comparison of Fecal Coliform Agar and Violet Red Bile Lactose Agar for Fecal Coliform Enumeration in Foods

Applied and Environmental Microbiology ◽

10.1128/aem.68.4.1631-1638.2002 ◽

2002 ◽

Vol 68 (4) ◽

pp. 1631-1638 ◽

Cited By ~ 63

Author(s):

A. Leclercq ◽

C. Wanegue ◽

P. Baylac

Keyword(s):

Escherichia Coli ◽

Fecal Coliform ◽

Food Samples ◽

Fecal Coliforms ◽

Predictive Values ◽

Direct Plating ◽

Hafnia Alvei ◽

E Coli ◽

Plating Method ◽

Mashed Potatoes

ABSTRACT A 24-h direct plating method for fecal coliform enumeration with a resuscitation step (preincubation for 2 h at 37 ± 1°C and transfer to 44 ± 1°C for 22 h) using fecal coliform agar (FCA) was compared with the 24-h standardized violet red bile lactose agar (VRBL) method. FCA and VRBL have equivalent specificities and sensitivities, except for lactose-positive non-fecal coliforms such as Hafnia alvei, which could form typical colonies on FCA and VRBL. Recovery of cold-stressed Escherichia coli in mashed potatoes on FCA was about 1 log unit lower than that with VRBL. When the FCA method was compared with standard VRBL for enumeration of fecal coliforms, based on counting carried out on 170 different food samples, results were not significantly different (P > 0.05). Based on 203 typical identified colonies selected as found on VRBL and FCA, the latter medium appears to allow the enumeration of more true fecal coliforms and has higher performance in certain ways (specificity, sensitivity, and negative and positive predictive values) than VRBL. Most colonies clearly identified on both media were E. coli and H. alvei, a non-fecal coliform. Therefore, the replacement of fecal coliform enumeration by E. coli enumeration to estimate food sanitary quality should be recommended.

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Introduction to the Food Safety Concerns of Verotoxin-Producing Escherichia coli

Experimental Biology and Medicine ◽

10.1177/153537020322800401 ◽

2003 ◽

Vol 228 (4) ◽

pp. 331-332 ◽

Cited By ~ 4

Author(s):

Hussein S. Hussein ◽

Stanley T. Omaye

Keyword(s):

Escherichia Coli ◽

Food Safety ◽

Safety Concern ◽

Critical Evaluation ◽

The United States ◽

Global Perspective ◽

E Coli ◽

The Past ◽

Food Borne Pathogens ◽

Food Borne

Verotoxin-producing Escherichia coli (VTEC) have emerged in the past two decades as food-borne pathogens that can cause major outbreaks of human illnesses worldwide. The number of outbreaks has increased in recent years due to changes in food production and processing systems, eating habits, microbial adaptation, and methods of VTEC transmission. The human illnesses range from mild diarrhea to hemolytic uremic syndrome (HUS) that can lead to death. The VTEC outbreaks have been attributed to O157:H7 and non-O157:H7 serotypes of E. coli. These E. coli serotypes include motile (e.g., O26:H11 and O104:H21) and nonmotile (e.g., O111:H–,0145:H–, and O157:H–) strains. In the United States, E. coli O157:H7 has been the major cause of VTEC outbreaks. Worldwide, however, non-O157:H7 VTEC (e.g., members of the 026, O103, O111, O118, O145, and O166 serogroups) have caused approximately 30% of the HUS cases in the past decade. Because large numbers of the VTEC outbreaks have been attributed to consumption of ruminant products (e.g., ground beef), cattle and sheep are considered reservoirs of these food-borne pathogens. Because of the food safety concern of VTEC, a global perspective on this problem is addressed (Exp Biol Med Vol. 228, No. 4). The first objective was to evaluate the known non-O157:H7 VTEC strains and the limitations associated with their detection and characterization. The second objective was to identify the VTEC serotypes associated with outbreaks of human illnesses and to provide critical evaluation of their virulence. The third objective was to determine the rumen effect on survival of E. coli O157:H7 as a VTEC model. The fourth objective was to explore the role of intimins in promoting attaching and effacing lesions in humans. Finally, the ability of VTEC to cause persistent infections in cattle was evaluated.

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Induction of Viable but Nonculturable Escherichia coli O157:H7 in the Phyllosphere of Lettuce: a Food Safety Risk Factor

Applied and Environmental Microbiology ◽

10.1128/aem.05020-11 ◽

2011 ◽

Vol 77 (23) ◽

pp. 8295-8302 ◽

Cited By ~ 81

Author(s):

Laura-Dorina Dinu ◽

Susan Bach

Keyword(s):

Escherichia Coli ◽

Food Safety ◽

Low Temperature ◽

Cell Density ◽

Prolonged Storage ◽

Potential Hazard ◽

Vbnc State ◽

Viable But Nonculturable ◽

Content Type ◽

E Coli

ABSTRACTEscherichia coliO157:H7 continues to be an important human pathogen and has been increasingly linked to food-borne illness associated with fresh produce, particularly leafy greens. The aim of this work was to investigate the fate ofE. coliO157:H7 on the phyllosphere of lettuce under low temperature and to evaluate the potential hazard of viable but nonculturable (VBNC) cells induced under such stressful conditions. First, we studied the survival of six bacterial strains following prolonged storage in water at low temperature (4°C) and selected two strains with different nonculturable responses for the construction ofE. coliO157:H7 Tn7gfptransformants in order to quantitatively assess the occurrence of human pathogens on the plant surface. Under a suboptimal growth temperature (16°C), bothE. coliO157:H7 strains maintained culturability on lettuce leaves, but under more stressful conditions (8°C), the bacterial populations evolved toward the VBNC state. The strain-dependent nonculturable response was more evident in the experiments with different inoculum doses (109and 106E. coliO157:H7 bacteria per g of leaf) when strain BRMSID 188 lost culturability after 15 days and strain ATCC 43895 lost culturability within 7 days, regardless of the inoculum dose. However, the number of cells entering the VBNC state in high-cell-density inoculum (approximately 55%) was lower than in low-cell-density inoculum (approximately 70%). We recorded the presence of verotoxin for 3 days in samples that contained a VBNC population of 4 to 5 log10cells but did not detect culturable cells. These findings indicate thatE. coliO157:H7 VBNC cells are induced on lettuce plants, and this may have implications regarding food safety.

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Multiplex Quantitative PCR Assays for the Detection and Quantification of the Six Major Non-O157 Escherichia coli Serogroups in Cattle Feces†

Journal of Food Protection ◽

10.4315/0362-028x.jfp-15-319 ◽

2016 ◽

Vol 79 (1) ◽

pp. 66-74 ◽

Cited By ~ 14

Author(s):

P. B. SHRIDHAR ◽

L. W. NOLL ◽

X. SHI ◽

B. AN ◽

N. CERNICCHIARO ◽

...

Keyword(s):

Escherichia Coli ◽

Quantitative Pcr ◽

Foodborne Pathogens ◽

Virulence Genes ◽

Target Genes ◽

Culture Methods ◽

Fecal Samples ◽

E Coli ◽

Cattle Feces ◽

Before And After

ABSTRACT Shiga toxin–producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, called non-O157 STEC, are important foodborne pathogens. Cattle, a major reservoir, harbor the organisms in the hindgut and shed them in the feces. Although limited data exist on fecal shedding, concentrations of non-O157 STEC in feces have not been reported. The objectives of our study were (i) to develop and validate two multiplex quantitative PCR (mqPCR) assays, targeting O-antigen genes of O26, O103, and O111 (mqPCR-1) and O45, O121, and O145 (mqPCR-2); (ii) to utilize the two assays, together with a previously developed four-plex qPCR assay (mqPCR-3) targeting the O157 antigen and three virulence genes (stx1, stx2, and eae), to quantify seven serogroups and three virulence genes in cattle feces; and (iii) to compare the three mqPCR assays to a 10-plex conventional PCR (cPCR) targeting seven serogroups and three virulence genes and culture methods to detect seven E. coli serogroups in cattle feces. The two mqPCR assays (1 and 2) were shown to be specific to the target genes, and the detection limits were 4 and 2 log CFU/g of pure culture–spiked fecal samples, before and after enrichment, respectively. A total of 576 fecal samples collected from a feedlot were enriched in E. coli broth and were subjected to quantification (before enrichment) and detection (after enrichment). Of the 576 fecal samples subjected, before enrichment, to three mqPCR assays for quantification, 175 (30.4%) were quantifiable (≥4 log CFU/g) for at least one of the seven serogroups, with O157 being the most common serogroup. The three mqPCR assays detected higher proportions of postenriched fecal samples (P < 0.01) as positive for one or more serogroups compared with cPCR and culture methods. This is the first study to assess the applicability of qPCR assays to detect and quantify six non-O157 serogroups in cattle feces and to generate data on fecal concentration of the six serogroups.

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Long-Haul Transport and Lack of Preconditioning Increases Fecal Shedding of Escherichia coli and Escherichia coli O157:H7 by Calves†

Journal of Food Protection ◽

10.4315/0362-028x-67.4.672 ◽

2004 ◽

Vol 67 (4) ◽

pp. 672-678 ◽

Cited By ~ 49

Author(s):

S. J. BACH ◽

T. A. McALLISTER ◽

G. J. MEARS ◽

K. S. SCHWARTZKOPF-GENSWEIN

Keyword(s):

Escherichia Coli ◽

Escherichia Coli O157 ◽

Fecal Samples ◽

Fecal Shedding ◽

E Coli ◽

Susceptibility To Infection

The effects of weaning and transport on fecal shedding of Escherichia coli and on E. coli O157:H7 were investigated using 80 Angus and 94 Charolais range steer calves blocked by breed and assigned to four treatments. The calves were or were not preconditioned before transport on commercial cattle liner to the feedlot via long (15 h) or short (3 h) hauling duration, yielding preconditioned long haul (P-L; n = 44), preconditioned short haul (P-S; n = 44), nonpreconditioned long haul (NP-L; n = 43), and nonpreconditioned short haul (NP-S; n = 43). Preconditioned calves were vaccinated and weaned 29 and 13 days, respectively, before transport. Nonpreconditioned calves were weaned 1 day before long or short hauling, penned for 24 h and hauled again for 2 h, and vaccinated on arrival at the feedlot. Fecal samples were collected from calves while on pasture, at weaning, at loading for transport, on arrival at the feedlot, twice in the first week, and on days 7, 14, 21, and 28 for enumeration of total E. coli (biotype 1) and detection of E. coli O157:H7. No calves were positive for E. coli O157:H7 before transport. Following transport, more (P < 0.005) NP-L calves (6 of 43) tested positive for E. coli O157:H7 than did P-L (1 of 44), NP-S (1 of 43), or P-S (0 of 44) calves, and on days 0, 1, 7, and 21, their levels of shedding of E. coli were higher (P < 0.005). The calves' susceptibility to infection from the environment (possibly the holding facilities or feedlot pens) was likely elevated by the stresses of weaning, transport, and relocation. Lack of preconditioning and long periods of transport (NP-L) increased fecal shedding of E. coli and E. coli O157:H7. Preconditioning may serve to reduce E. coli O157:H7 shedding by range calves on arrival at the feedlot.

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