James E. Wells

Prevalence and level of Escherichia coli O157:H7 in feces and on hides of feedlot steers fed diets with or without wet distillers grains with solubles.

The objective of this study was to determine if wet distillers grains with solubles (WDGS) from corn in diets affected Escherichia coli O157:H7 in growing and finishing cattle; steers (n = 603) were randomly assigned to diets with or without WDGS. Hide and fecal samples were collected monthly (October through June) from each animal for enumeration and enrichment of E. coli O157:H7. In the growing phase (0 or 13.9% WDGS diets), fecal prevalence for E. coli O157:H7 in steers fed a diet with WDGS was twice that of the prevalence in control steers (P < 0.001). In the finishing phase (0 or 40% WDGS diets), the average prevalence in feces (P < 0.001) and on hides (P < 0.001) was higher for cattle fed WDGS. The average percentage of fecal E. coli O157:H7 enumerable samples during the finishing phase for cattle fed WDGS was 2.7% compared with 0.1% for control steers (P < 0.001). The average percentage of E. coli O157:H7 enumerable hide samples was not different between diets, but the cattle fed WDGS had higher levels (P < 0.05) of the pathogen. Animals fed WDGS had higher levels of E. coli (P < 0.001), higher pH values (P < 0.001), and lower concentrations of L-lactate (P < 0.001) in feces than those values of the control steers. These results indicate that feeding 40% WDGS could increase the level and prevalence of E. coli O157:H7 in and on feedlot cattle when E. coli O157:H7 is seasonally low.

Prevalence and Characterization of Salmonella in Bovine Lymph Nodes Potentially Destined for Use in Ground Beef

A potential source of pathogenic bacteria in ground beef is the lymphatic system, specifically the lymph nodes. Bacteria have been isolated from the lymph nodes of cattle at slaughter; however, most studies have dealt with mesenteric lymph nodes, which are not normally incorporated into ground beef. The objective of the current study was to determine the prevalence and multidrug-resistance status of Salmonella in bovine lymph nodes associated with lean and fat trimmings that might be utilized in ground beef production. Bovine lymph nodes (n = 1,140) were collected from commercial beef processing plants. Half of the lymph nodes sampled were obtained from cull cow and bull processing plants, and the remainder were obtained from fed beef processing plants. Lymph nodes located in chuck and flank adipose tissue were collected for this study. Salmonella prevalence in the lymph node samples was low, with an overall prevalence of 1.6% and a 95% confidence interval of 0.85 to 2.3%. Lymph nodes from cull cattle carcasses had a higher prevalence of Salmonella than did those from fed cattle carcasses. Lymph nodes from the flanks of cow and bull carcasses had the highest prevalence at 3.86%, whereas lymph nodes from the chuck region of fed cattle carcasses had the lowest prevalence at 0.35%. Three of the 18 Salmonella-positive lymph node samples contained multidrug-resistant Salmonella, and all 3 samples were from cull cattle.

Escherichia coli O157:H7: recent advances in research on occurrence, transmission, and control in cattle and the production environment.

Escherichia coli O157:H7 is a zoonotic pathogen that is an important cause of human foodborne and waterborne disease, with a spectrum of illnesses ranging from asymptomatic carriage and diarrhea to the sometimes fatal hemolytic uremic syndrome. Outbreaks of E. coli O157:H7 disease are often associated with undercooked beef, but there are other sources of transmission, including water, produce, and animal contact, which can often be linked directly or indirectly to cattle. Thus, preharvest control of this pathogen in cattle production should have a large impact on reducing the risk of human foodborne illness. In this review, we will summarize preharvest research on E. coli O157:H7 in cattle and the production environment, focusing on factors that may influence the transmission, prevalence, and levels of this pathogen, such as season, diet, high-level shedders, and animal stress. In addition, we will discuss recent research on the reduction of this pathogen in cattle production, including vaccination, probiotics, bacteriophage, and manure treatments.

Chromogenic agar medium for detection and isolation of Escherichia coli serogroups O26, O45, O103, O111, O121, and O145 from fresh beef and cattle feces.

Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are clinically important foodborne pathogens. Unlike E. coli O157:H7, these foodborne pathogens have no unique biochemical characteristics to readily distinguish them from other E. coli strains growing on plating media. In this study, a chromogenic agar medium was developed in order to differentiate among non-O157 STEC strains of serogroups O26, O45, O103, O111, O121, and O145 on a single agar medium. The ability of this chromogenic agar medium to select and distinguish among these pathogens is based on a combination of utilization of carbohydrates, b -galactosidase activity, and resistance to selective agents. The agar medium in combination with immunomagnetic separation was evaluated and successfully allowed for the detection and isolation of these six serogroups from artificially contaminated fresh beef. The agar medium in combination with immunomagnetic separation also allowed successful detection and isolation of naturally occurring non-O157 STEC strains present in cattle feces. Thirty-five strains of the top six non-O157 STEC serogroups were isolated from 1,897 fecal samples collected from 271 feedlot cattle. This chromogenic agar medium could help significantly in routine screening for the top six non-O157 STEC serogroups from beef cattle and other food.

Evaluation of a Direct-Fed Microbial Product Effect on the Prevalence and Load of Escherichia coli O157:H7 in Feedlot Cattle†

Direct-fed microbials (DFM) have been identified as potential preharvest interventions for the reduction of foodborne bacterial pathogens such as Escherichia coli O157:H7. This study evaluated the efficacy of a DFM consisting of Bacillus subtilis strain 166 as an antimicrobial intervention strategy for the reduction of prevalence and load of E. coli O157:H7 in feces and on hides of feedlot cattle. Cattle (n = 526) were divided among 16 feedlot pens. Half of the pens received the DFM, and the other half did not. Hide and fecal samples were collected from each animal on days 28, 63, and 84 of the feeding trial. Over the course of the 84-day feeding period, there were no significant differences observed between treatments for either hide or fecal prevalence of E. coli O157:H7, or for the percentage of animals that were shedding E. coli O157:H7 at high levels (> or =200 CFU/g) in their feces or harboring E. coli O157:H7 at high levels (> or =40 CFU/cm(2)) on their hides. In addition, there was no significant difference between the average daily gains for the treated and control groups, with both groups averaging 1.3 kg/day. We concluded that the DFM tested would not be an effective preharvest intervention against E. coli O157:H7.

Fate of naturally occurring Escherichia coli O157:H7 and other zoonotic pathogens during minimally managed bovine feedlot manure composting processes.

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 m(3) 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.

Soil versus pond ash surfacing of feedlot pens: occurrence of Escherichia coli O157:H7 in cattle and persistence in manure.

Reducing Escherichia coli O157:H7 in cattle and their manure is critical for reducing the risk for human foodborne and waterborne illness. The objective of this study was to evaluate the effects of soil and pond ash surfaces for feedlot pens on the prevalence, levels, and/or persistence of naturally occurring E. coli O157:H7 and total E. coli in cattle (feces and hides) and manure. Cattle (128 beef heifers) were sorted among 16 pens: 8 surfaced with soil and 8 surfaced with pond ash. The prevalence of E. coli O157:H7 in feces decreased (P < 0.0001) during the study from 57.0% on day 0 to 3.9% on day 84 but did not differ (P > or = 0.05) between cattle on soil and on pond ash pens at any sampling period. The prevalence of the pathogen on hides and in feedlot surface material (FSM) also decreased (P < 0.0001), with no effect of soil or pond ash surface (P > or = 0.05). Similarly, levels of E. coli in FSM did not differ (P > or = 0.05) at any sampling period, and there were no clear trends for survival differences of E. coli O157:H7 or E. coli in FSM between pond ash and soil surfaces, although E. coli populations survived at 5.0 log CFU/g of FSM on the pen surfaces 6 weeks after the cattle were removed. These results indicate that housing cattle on pens surfaced with pond ash versus pens surfaced with soil does not affect E. coli O157:H7 in cattle or their manure.

Prevalence of Mycobacterium avium subsp. paratuberculosis in ileocecal lymph nodes and on hides and carcasses from cull cows and fed cattle at commercial beef processing plants in the United States.

Clinical associations between Crohns disease in humans and Mycobacterium avium subsp. paratuberculosis (MAP) have been suggested but not confirmed. Cattle could be sources for MAP, but little information on MAP prevalence with beef has been reported. Samples of ileocecal lymph nodes and swabs of hides and carcasses from 343 animals at cull cattle slaughtering facilities and 243 animals at fed cattle slaughtering facilities across the United States were analyzed for the presence of MAP. Amplification of genetic sequences detected MAP DNA predominantly on hides and in lymph nodes of samples taken at both types of processing facilities. More than 34% of the cattle at cull cow slaughtering facilities had ileocecal lymph nodes that tested positive for MAP DNA. From these same cattle, hide prevalence was more than twofold greater than the prevalence in ileocecal lymph nodes, suggesting that cross-contamination could be occurring during transport and lairage. The prevalence of MAP DNA decreased during processing, and less than 11% of the carcasses tested positive after interventions in the cull cow processing facilities. Using standard double-decontamination and culture techniques, less than 1% of the postintervention carcasses tested positive for viable MAP at cull cow facilities. In samples from the facilities processing only fed cattle, MAP prevalence of 1% or less was detected for ileocecal lymph node, hide, and carcass samples, and viable MAP was not detected. Based on this study, fed cattle carcasses are unlikely sources of MAP, and carcasses at cull cow plants have only a slight risk for transmitting viable MAP, due to current interventions.

Impact of reducing the level of wet distillers grains fed to cattle prior to harvest on prevalence and levels of Escherichia coli O157:H7 in feces and on hides.

Cattle fed finishing diets with wet distillers grains with solubles (WDGS) have been shown to harbor increased Escherichia coli O157:H7 populations in the feces and on the hides. To determine if feeding a lower level of WDGS at the end of the feeding period reduces E. coli O157:H7 load at harvest, 608 heifers were sorted into one of five treatments and fed 0, 40, or 70% WDGS (dry matter basis). For three of the treatments, WDGS was reduced midway through the study. Treatment 0W0W heifers (positive control) were fed a corn grain-based diet continuously, and 40W40W heifers (negative control) were fed 40% WDGS continuously. Heifers subjected to treatments 40W0W, 40W15W, and 70W15W were fed either 40 or 70% WDGS for the first 56 days and switched to 0 or 15% WDGS, respectively, for the last 56 days. Prior to the switch in diets, animals fed diets with 40 or 70% had higher prevalence and percent enumerable fecal samples for E. coli O157:H7. After the dietary switch, animals fed 40W0W, 40W15W, and 70W15W diets had fecal prevalence and percent enumerable samples (33.4 and 6.3%, 31.0 and 9.7%, and 34.9 and 8.4%, respectively) similar to those of animals fed 0W0W diets (10.2 and 3.2%, respectively; P > 0.05), whereas animals fed 40W40W had the highest fecal prevalence and percent enumerable samples (70.1 and 29.2%, respectively; P < 0.05). Similar relationships between the treatments were observed for hide samples. Time after dietary switch was important, as animals fed lower levels had significantly lower fecal prevalence and percent enumerable samples after 56 days, but not after 28 days. The study indicates that cattle can be switched to lower levels of dietary WDGS (15% or less) 56 days prior to harvest to significantly reduce E. coli O157:H7 in feces and on hides.

Impacts of Individual Animal Response to Heat and Handling Stresses on Escherichia coli and E. coli O157:H7 Fecal Shedding by Feedlot Cattle

The reduction of foodborne pathogens in cattle destined for human consumption will require knowledge of the factors that impact the carriage and shedding of these organisms. The objective of this work was to investigate the effects of heat and handling stress levels on the fecal shedding of Escherichia coli O157:H7 and generic E. coli by feedlot cattle. In year 1, 128 feedlot heifers were evaluated for heat tolerance five times per week during the 84-day finishing period from May through August. Heat stress measurements included respiration rate, panting score, and visual assessments. In year 2, panting scores were taken for a group of 256 finishing feedlot heifers on days in July and August for which the temperature humidity index (THI) was predicted to be in the emergency category (THI > or = 84). For both years, animals were weighed and temperament scored to assess handling stress on a 28-day schedule. At the same time, rectal fecal samples were collected from each animal individually. The presence and concentrations of E. coli O157:H7 and concentrations of generic E. coli in feces were determined. There were no clear trends between the heat stress levels or temperament scores (as an indicator of response to handling) with either fecal generic E. coli concentrations or E. coli O157:H7 concentrations or prevalence in feces, indicating that neither heat nor handling stress contributes to the food safety risk associated with E. coli O157:H7-positive cattle.