scholarly journals Escherichia coli O157:H7 in Environments of Culture-Positive Cattle

2005 ◽  
Vol 71 (11) ◽  
pp. 6816-6822 ◽  
Author(s):  
Margaret A. Davis ◽  
Karen A. Cloud-Hansen ◽  
John Carpenter ◽  
Carolyn J. Hovde
Keyword(s):  
Escherichia Coli ◽  
Room Temperature ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Different Temperatures ◽  
Farm Structures ◽  
On Farm ◽  
Culture Positive ◽  
Culture Negative ◽  
Over Time

ABSTRACT Outbreaks of Escherichia coli O157:H7 disease associated with animal exhibits have been reported with increasing frequency. Transmission can occur through contact with contaminated haircoats, bedding, farm structures, or water. We investigated the distribution and survival of E. coli O157:H7 in the immediate environments of individually housed, experimentally inoculated cattle by systematically culturing feed, bedding, water, haircoat, and feed bunk walls for E. coli O157:H7 for 3 months. Cedar chip bedding was the most frequently culture-positive environmental sample tested (27/96 or 28.15%). Among these, 12 (44.0%) of positive bedding samples were collected when the penned animal was fecal culture negative. Survival of E. coli O157:H7 in experimentally inoculated cedar chip bedding and in grass hay feed was determined at different temperatures. Survival was longest in feed at room temperature (60 days), but bacterial counts decreased over time. The possibility that urine plays a role in the environmental survival of E. coli O157:H7 was investigated. Cedar chip bedding moistened with sterile water or bovine urine was inoculated with E. coli O157:H7. Bedding moistened with urine supported growth of E. coli O157:H7, whereas inoculated bedding moistened with only water yielded decreasing numbers of bacteria over time. The findings that environmental samples were frequently positive for E. coli O157:H7 at times when animals were culture negative and that urine provided a substrate for E. coli O157:H7 growth have implications for understanding the on-farm ecology of this pathogen and for the safety of ruminant animal exhibits, particularly petting zoos and farms where children may enter animal pens.

scholarly journals Performance of the ImmunoCard STAT! E. coli O157:H7 Test for Detection of Escherichia coli O157:H7 in Stools

2000 ◽  
Vol 38 (5) ◽  
pp. 1866-1868 ◽  
Author(s):  
Andrew Mackenzie ◽  
Elaine Orrbine ◽  
Lucie Hyde ◽  
Michelle Benoit ◽  
Frank Chan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Sensitivity ◽  
False Positives ◽  
Broth Culture ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Stool Specimens ◽  
Multicenter Prospective Study ◽  
Culture Positive ◽  
Culture Negative

ImmunoCard STAT! E. coli O157:H7 (Meridian Diagnostics, Inc., Cincinnati, Ohio) is a novel rapid (10-min) test for the presence of Escherichia coli O157:H7 in stools. The test may be performed either directly on stool specimens or on an overnight broth culture of stool. In a multicenter prospective study, 14 of 14 specimens positive by culture for E. coli O157:H7 were positive by the ImmunoCard STAT! O157:H7 test, and there were no false positives from 263 culture-negative specimens. In a retrospective study, the test was positive in 339 (81%) of 417 stored culture-positive specimens and the specificity was 95% (98 of 103 specimens). No false positives were associated with alternate stool pathogens. The ImmunoCard STAT! O157:H7 test has high sensitivity and specificity.

Validation of Time and Temperature Values as Critical Limits for the Control of Escherichia coli O157:H7 during the Production of Fresh Ground Beef

2006 ◽  
Vol 69 (8) ◽  
pp. 1978-1982 ◽  
Author(s):  
J. E. MANN ◽  
M. M. BRASHEARS
Keyword(s):  
Escherichia Coli ◽  
Room Temperature ◽  
Hazard Analysis ◽  
Control Point ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Meat Processing ◽  
E Coli ◽  
Critical Control Point ◽  
Critical Limits

In order to provide beef processors with valuable data to validate critical limits set for temperature during grinding, a study was conducted to determine Escherichia coli O157:H7 growth at various temperatures in raw ground beef. Fresh ground beef samples were inoculated with a cocktail mixture of streptomycin-resistant E. coli O157:H7 to facilitate recovery in the presence of background flora. Samples were held at 4.4, 7.2, and 10°C, and at room temperature (22.2 to 23.3°C) to mimic typical processing and holding temperatures observed in meat processing environments. E. coli O157:H7 counts were determined by direct plating onto tryptic soy agar with streptomycin (1,000 μg/ml), at 2-h intervals over 12 h for samples held at room temperature. Samples held under refrigeration temperatures were sampled at 4, 8, 12, 24, 48, and 72 h. Less than one log of E. coli O157:H7 growth was observed at 48 h for samples held at 10°C. Samples held at 4.4 and 7.2°C showed less than one log of E. coli O157:H7 growth at 72 h. Samples held at room temperature showed no significant increase in E. coli O157:H7 counts for the first 6 h, but increased significantly afterwards. These results illustrate that meat processors can utilize a variety of time and temperature combinations as critical limits in their hazard analysis critical control point plans to minimize E. coli O157:H7 growth during the production and storage of ground beef.

Monitoring Escherichia coli O157:H7 in Inoculated and Naturally Colonized Feedlot Cattle and Their Environment

2005 ◽  
Vol 68 (1) ◽  
pp. 26-33 ◽  
Author(s):  
K. STANFORD ◽  
S. J. BACH ◽  
T. H. MARX ◽  
S. JONES ◽  
J. R. HANSEN ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Feedlot Cattle ◽  
Mitigation Strategies ◽  
Escherichia Coli O157 ◽  
Feeding Period ◽  
Sample Collection ◽  
E Coli ◽  
Challenge Study ◽  
On Farm ◽  
Feed Samples

On-farm methods of monitoring Escherichia coli O157:H7 were assessed in 30 experimentally inoculated steers housed in four pens over a 12-week period and in 202,878 naturally colonized feedlot cattle housed in 1,160 pens on four commercial Alberta feedlots over a 1-year period. In the challenge study, yearling steers were experimentally inoculated with 1010 CFU of a four-strain mixture of nalidixic acid–resistant E. coli O157:H7. After inoculation, shedding of E. coli O157:H7 was monitored weekly by collecting rectal fecal samples (FEC), oral swabs (ORL), pooled fecal pats (PAT), manila ropes (ROP) orally accessed for 4 h, feed samples, water, and water bowl interface. Collection of FEC from all animals per pen provided superior isolation (P < 0.01) of E. coli O157:H7 compared with other methods, although labor and animal restraint requirements for fecal sample collection were high. When one sample was collected per pen of animals, E. coli O157:H7 was more likely to be detected from the ROP than from the FEC, PAT, or ORL (P < 0.001). In the commercial feedlot study, samples were limited to ROP and PAT, and E. coli O157:H7 was isolated in 18.8% of PAT and 6.8% of ROP samples. However, for animals that had been resident in the feedlot pen for at least 1 month, isolation of E. coli O157:H7 from ROP was not different from that from PAT (P = 0.35). Pens of animals on feed for <30 days were six times more likely to shed E. coli O157:H7 than were animals on feed for >30 days. However, change in diet did not affect shedding of the organism (P > 0.23) provided that animals had acclimated to the feedlot for 1 month or longer. Findings from this study indicate the importance of introduction of mitigation strategies early in the feeding period to reduce transference and the degree to which E. coli O157:H7 is shed into the environment.

Escherichia coli O157 in Cull Dairy Cows on Farm and at Slaughter

1997 ◽  
Vol 60 (11) ◽  
pp. 1386-1387 ◽  
Author(s):  
DANIEL H. RICE ◽  
ERIC D. EBEL ◽  
DALE D. HANCOCK ◽  
THOMAS E. BESSER ◽  
DONALD E. HERRIOTT ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Dairy Cattle ◽  
Dairy Cows ◽  
Dairy Herds ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Cull Cows ◽  
On Farm

Cull dairy cattle both on the farm and at slaughter from herds in the states of Idaho, Oregon, and Washington were surveyed for Escherichia coli O157 by culturing fecal swab samples. A total of 205 cull cows from 19 dairy herds were sampled on the farm of origin; 7 (3.4%) tested positive for E. coli O157. A total of 103 cull cows from 15 dairy herds were sampled at slaughter; 4 (3.9%) were positive for E. coli O157. Eighty-nine cull cows were sampled both at the farm and at slaughter; 2 (2.2%) were positive in both locations, 3 (3.3%) only on the farm, and 2 (2.2%) only at the slaughter plant. Seven (7.9%) of the 89 cull cows tracked from farm to slaughter were positive in at least one location. This suggests a higher prevalence of E. coli O157 in cull dairy cattle than previously has been reported to occur in other ages and classes of cattle.

Effect of Route of Introduction and Host Cultivar on the Colonization, Internalization, and Movement of the Human Pathogen Escherichia coli O157:H7 in Spinach

2009 ◽  
Vol 72 (7) ◽  
pp. 1521-1530 ◽  
Author(s):  
R. MITRA ◽  
E. CUESTA-ALONSO ◽  
A. WAYADANDE ◽  
J. TALLEY ◽  
S. GILLILAND ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fluorescent Protein ◽  
Sprinkler Irrigation ◽  
Conclusive Evidence ◽  
Escherichia Coli O157 ◽  
Human Pathogens ◽  
Soil Infiltration ◽  
Fresh Tissue ◽  
E Coli ◽  
Over Time

Human pathogens can contaminate leafy produce in the field by various routes. We hypothesized that interactions between Escherichia coli O157:H7 and spinach are influenced by the route of introduction and the leaf microenvironment. E. coli O157:H7 labeled with green fluorescent protein was dropped onto spinach leaf surfaces, simulating bacteria-laden raindrops or sprinkler irrigation, and survived on the phylloplane for at least 14 days, with increasing titers and areas of colonization over time. The same strains placed into the rhizosphere by soil infiltration remained detectable on very few plants and in low numbers (102 to 106 CFU/g fresh tissue) that decreased over time. Stem puncture inoculations, simulating natural wounding, rarely resulted in colonization or multiplication. Bacteria forced into the leaf interior survived for at least 14 days in intercellular spaces but did not translocate or multiply. Three spinach cultivars with different leaf surface morphologies were compared for colonization by E. coli O157:H7 introduced by leaf drop or soil drench. After 2 weeks, cv. Bordeaux hosted very few bacteria. More bacteria were seen on cv. Space and were dispersed over an area of up to 0.3 mm2. The highest bacterial numbers were observed on cv. Tyee but were dispersed only up to 0.15 mm2, suggesting that cv. Tyee may provide protected niches or more nutrients or may promote stronger bacterial adherence. These findings suggest that the spinach phylloplane is a supportive niche for E. coli O157:H7, but no conclusive evidence was found for natural entry into the plant interior. The results are relevant for interventions aimed at minimizing produce contamination by human pathogens.

Comparing Real-Time and Conventional PCR to Culture-Based Methods for Detecting and Quantifying Escherichia coli O157 in Cattle Feces

2014 ◽  
Vol 77 (2) ◽  
pp. 314-319 ◽  
Author(s):  
M. E. JACOB ◽  
J. BAI ◽  
D. G. RENTER ◽  
A. T. ROGERS ◽  
X. SHI ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Positive Culture ◽  
Feedlot Cattle ◽  
Escherichia Coli O157 ◽  
Conventional Pcr ◽  
Culture Methods ◽  
E Coli ◽  
Cattle Feces ◽  
Culture Negative

Detection of Escherichia coli O157 in cattle feces has traditionally used culture-based methods; PCR-based methods have been suggested as an alternative. We aimed to determine if multiplex real-time (mq) or conventional PCR methods could reliably detect cattle naturally shedding high (≥104 CFU/g of feces) and low (~102 CFU/g of feces) concentrations of E. coli O157. Feces were collected from pens of feedlot cattle and evaluated for E. coli O157 by culture methods. Samples were categorized as (i) high shedders, (ii) immunomagnetic separation (IMS) positive after enrichment, or (iii) culture negative. DNA was extracted pre- and postenrichment from 100 fecal samples from each category (high shedder, IMS positive, culture negative) and subjected to mqPCR and conventional PCR assays based on detecting three genes, rfbE, stx1, and stx2. In feces from cattle determined to be E. coli O157 high shedders by culture, 37% were positive by mqPCR prior to enrichment; 85% of samples were positive after enrichment. In IMS-positive samples, 4% were positive by mqPCR prior to enrichment, while 43% were positive after enrichment. In culture-negative feces, 7% were positive by mqPCR prior to enrichment, and 40% were positive after enrichment. The proportion of high shedder–positive and culture-positive (high shedder and IMS) samples were significantly different from mqPCR-positive samples before and after enrichment (P < 0.01). Similar results were observed for conventional PCR. Our data suggest that mqPCR and conventional PCR are most useful in identifying high shedder animals and may not be an appropriate substitute to culture-based methods for detection of E. coli O157 in cattle feces.

scholarly journals Effect of Forage or Grain Diets with or without Monensin on Ruminal Persistence and Fecal Escherichia coli O157:H7 in Cattle

2004 ◽  
Vol 70 (9) ◽  
pp. 5336-5342 ◽  
Author(s):  
M. J. Van Baale ◽  
J. M. Sargeant ◽  
D. P. Gnad ◽  
B. M. DeBey ◽  
K. F. Lechtenberg ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Nalidixic Acid ◽  
Escherichia Coli O157 ◽  
Fecal Samples ◽  
Fecal Shedding ◽  
E Coli ◽  
Culture Positive ◽  
Forage Diets

ABSTRACT Twelve ruminally cannulated cattle, adapted to forage or grain diet with or without monensin, were used to investigate the effects of diet and monensin on concentration and duration of ruminal persistence and fecal shedding of E. coli O157:H7. Cattle were ruminally inoculated with a strain of E. coli O157:H7 (1010 CFU/animal) made resistant to nalidixic acid (Nalr). Ruminal and fecal samples were collected for 11 weeks, and then cattle were euthanized and necropsied and digesta from different gut locations were collected. Samples were cultured for detection and enumeration of Nalr E. coli O157:H7. Cattle fed forage diets were culture positive for E. coli O157:H7 in the feces for longer duration (P < 0.05) than cattle fed a grain diet. In forage-fed cattle, the duration they remained culture positive for E. coli O157:H7 was shorter (P < 0.05) when the diet included monensin. Generally, ruminal persistence of Nalr E. coli O157:H7 was not affected by diet or monensin. At necropsy, E. coli O157:H7 was detected in cecal and colonic digesta but not from the rumen. Our study showed that cattle fed a forage diet were culture positive longer and with higher numbers than cattle on a grain diet. Monensin supplementation decreased the duration of shedding with forage diet, and the cecum and colon were culture positive for E. coli O157:H7 more often than the rumen of cattle.

Fate of Naturally Occurring Escherichia coli O157:H7 and Other Zoonotic Pathogens during Minimally Managed Bovine Feedlot Manure Composting Processes†

2013 ◽  
Vol 76 (8) ◽  
pp. 1308-1321 ◽  
Author(s):  
ELAINE D. BERRY ◽  
PATRICIA D. MILLNER ◽  
JAMES E. WELLS ◽  
NORASAK KALCHAYANAND ◽  
MICHAEL N. GUERINI
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
High Temperatures ◽  
Thermophilic Bacteria ◽  
Foodborne Illness ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Naturally Occurring ◽  
On Farm ◽  
Campylobacter Spp

Reducing Escherichia coli O157:H7 in livestock manures before application to cropland is critical for reducing the risk of foodborne illness associated with produce. Our objective was to determine the fate of naturally occurring E. coli O157:H7 and other pathogens during minimally managed on-farm bovine manure composting processes. Feedlot pen samples were screened to identify E. coli O157:H7–positive manure. Using this manure, four piles of each of three different composting formats were constructed in each of two replicate trials. Composting formats were (i) turned piles of manure plus hay and straw, (ii) static stockpiles of manure, and (iii) static piles of covered manure plus hay and straw. Temperatures in the tops, toes, and centers of the conical piles (ca. 6.0 m3 each) were monitored. Compost piles that were turned every 2 weeks achieved higher temperatures for longer periods in the tops and centers than did piles that were left static. E. coli O157:H7 was not recovered from top samples of turned piles of manure plus hay and straw at day 28 and beyond, but top samples from static piles were positive for the pathogen up to day 42 (static manure stockpiles) and day 56 (static covered piles of manure plus hay and straw). Salmonella, Campylobacter spp., and Listeria monocytogenes were not found in top or toe samples at the end of the composting period, but E. coli O157:H7 and Listeria spp. were recovered from toe samples at day 84. Our findings indicate that some minimally managed composting processes can reduce E. coli O157:H7 and other pathogens in bovine manure but may be affected by season and/or initial levels of indigenous thermophilic bacteria. Our results also highlight the importance of adequate C:N formulation of initial mixtures for the production of high temperatures and rapid composting, and the need for periodic turning of the piles to increase the likelihood that all parts of the mass are subjected to high temperatures.

Real-Time Bioluminescence Analysis of Escherichia coli O157:H7 Survival on Livestock Meats Stored Fresh, Cold, or Frozen

2018 ◽  
Vol 81 (11) ◽  
pp. 1906-1912 ◽  
Author(s):  
SEONG B. PARK ◽  
SHECOYA B. WHITE ◽  
CHRISTY S. STEADMAN ◽  
CLAY A. CAVINDER ◽  
SCOTT T. WILLARD ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Bacterial Growth ◽  
Room Temperature ◽  
Storage Conditions ◽  
Escherichia Coli O157 ◽  
Production Environment ◽  
Real Time Monitoring ◽  
E Coli ◽  
C Storage

ABSTRACT Foodborne bacteria such as Escherichia coli O157:H7 can cause severe hemorrhagic colitis in humans following consumption of contaminated meat products. Contamination with pathogenic bacteria is frequently found in the food production environment, and adequate household storage conditions of purchased foods are vital for illness avoidance. Real-time monitoring was used to evaluate bacterial growth in ground horse, beef, and pork meats maintained under various storage conditions. Various levels of E. coli O157:H7 carrying the luxCDABE operon, which allows the cells to emit bioluminescence, were used to inoculate meat samples that were then stored at room temperature for 0.5 day, at 4°C (cold) for 7 or 9 days, or −20°C (frozen) for 9 days. Real-time bioluminescence imaging (BLI) of bacterial growth was used to assess bacterial survival or load. Ground horse meat BLI signals and E. coli levels were dose and time dependent, increasing during room temperature and −20°C storage, but stayed at low levels during 4°C storage. No bacteria survived in the lower level inoculum groups (101 and 103 CFU/g). With an inoculum of 107 CFU/g, pork meats had higher BLI signals than did their beef counterparts, displaying decreased BLI signals during 7 days storage at 4°C. Both meat types had higher BLI signals in the fat area, which was confirmed with isolated fat tissues in the beef meat. Beef lean and fat tissues contrasted with both pork fat and lean tissues, which had significantly higher BLI signals and bacterial levels. BLI appears to be a useful research tool for real-time monitoring of bacterial growth and survival in various stored livestock meats. The dependence of E. coli O157:H7 growth on meat substrate (fat or lean) and storage conditions may be used as part of an effective antibacterial approach for the production of safe ground horse, beef, and pork meats.

Factors Impacting the Regrowth of Escherichia coli O157:H7 in Dairy Manure Compost

2009 ◽  
Vol 72 (7) ◽  
pp. 1576-1584 ◽  
Author(s):  
JINKYUNG KIM ◽  
FENG LUO ◽  
XIUPING JIANG
Keyword(s):  
Escherichia Coli ◽  
Moisture Content ◽  
Room Temperature ◽  
Dairy Manure ◽  
Population Increase ◽  
Escherichia Coli O157 ◽  
Inoculum Level ◽  
Factors Affecting ◽  
E Coli ◽  
Manure Compost

The environmental variables affecting Escherichia coli O157:H7 regrowth in dairy manure compost were investigated. Factors evaluated were moisture content, strain variation, growth medium of inoculum, level of background microflora and inoculum, different days of composting, and acclimation at room temperature. A mathematical model was applied to describe E. coli O157 regrowth potential in compost. Repopulation occurred in autoclaved compost with a moisture content as low as 20% (water activity of 0.986) in the presence of background microflora of 2.3 to 3.9 log CFU/g. The population of all three E. coli O157 strains increased from ca. 1 to 4.85 log CFU/g in autoclaved compost, with the highest increase in the spinach-outbreak strain. However, E. coli O157 regrowth was suppressed by background microflora at ca. 6.5 log CFU/g. By eliminating acclimation at room temperature and increasing the inoculum level to ca. 3 log CFU/g, E. coli O157:H7 could regrow in the presence of high levels of background microflora. E. coli O157:H7 regrowth in the autoclaved compost collected from the field study was evident at all sampling days, with the population increase ranging from 3.49 to 6.54 log CFU/g. The fate of E. coli O157:H7 in compost was well described by a Whiting and Cygnarowicz-Provost model, with R2 greater than 0.9. The level of background microflora was a significant factor for both growth and death parameters. Our results reveal that a small number of E. coli O157 cells can regrow in compost, and both background microflora and moisture content were major factors affecting E. coli O157:H7 growth.

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