Wide but Variable Distribution of a Hypervirulent Campylobacter jejuni Clone in Beef and Dairy Cattle in the United States.

Campylobacter jejuni clone SA is the major cause of sheep abortion and contributes significantly to foodborne illnesses in the United States. Clone SA is hypervirulent because of its distinct ability to produce systemic infection and its predominant role in clinical sheep abortion. Despite the importance of clone SA, little is known about its distribution and epidemiological features in cattle. Here we describe a prospective study on C. jejuni clone SA prevalence in 35 feedlots in 5 different states in the United States and a retrospective analysis of clone SA in C. jejuni isolates collected by National Animal Health Monitoring System (NAHMS) dairy studies in 2002, 2007, and 2014. In feedlot cattle feces, the overall prevalence of Campylobacter organisms was 72.2%, 82.1% of which were C. jejuni Clone SA accounted for 5.8% of the total C. jejuni isolates, but its prevalence varied by feedlot and state. Interestingly, starlings on the feedlots harbored C. jejuni in feces, including clone SA, suggesting that these birds may play a role in the transmission of Campylobacter In dairy cattle, the overall prevalence of clone SA was 7.2%, but a significant decrease in the prevalence was observed from 2002 to 2014. Whole-genome sequence analysis of the dairy clone SA isolates revealed that it was genetically stable over the years and most of the isolates carried the tetracycline resistance gene tet(O) in the chromosome. These findings indicate that clone SA is widely distributed in both beef and dairy cattle and provide new insights into the molecular epidemiology of clone SA in ruminants.IMPORTANCEC. jejuni clone SA is a major cause of small-ruminant abortion and an emerging threat to food safety because of its association with foodborne outbreaks. Cattle appear to serve as a major reservoir for this pathogenic organism, but there is a major gap in our knowledge about the epidemiology of clone SA in beef and dairy cattle. By taking advantage of surveillance studies conducted on a national scale, we found a wide but variable distribution of clone SA in feedlot cattle and dairy cows in the United States. Additionally, the work revealed important genomic features of clone SA isolates from cattle. These findings provide critically needed information for the development of preharvest interventions to control the transmission of this zoonotic pathogen. Control of C. jejuni clone SA will benefit both animal health and public health, as it is a zoonotic pathogen causing disease in both ruminants and humans.

Examining trust factors in online food risk information: The case of unpasteurized or 'raw' milk.

The internet has become an increasingly important way of communicating with consumers about food risk information. However, relatively little is known about how consumers evaluate and come to trust the information they encounter online. Using the example of unpasteurized or raw milk this paper presents two studies exploring the trust factors associated with online information about the risks and benefits of raw milk consumption. In the first study, eye-tracking data was collected from 33 pasteurised milk consumers whilst they viewed six different milk related websites. A descriptive analysis of the eye-tracking data was conducted to explore viewing patterns. Reports revealed the importance of images as a way of capturing initial attention and foregrounding other features and highlighted the significance of introductory text within a homepage. In the second, qualitative study, 41 consumers, some of whom drank raw milk, viewed a selection of milk related websites before participating in either a group discussion or interview. Seventeen of the participants also took part in a follow up telephone interview 2 weeks later. The qualitative data supports the importance of good design whilst noting that balance, authorship agenda, the nature of evidence and personal relevance were also key factors affecting consumers trust judgements. The results of both studies provide support for a staged approach to online trust in which consumers engage in a more rapid, heuristic assessment of a site before moving on to a more in-depth evaluation of the information available. Findings are discussed in relation to the development of trustworthy online food safety resources.

Subboiling Moist Heat Favors the Selection of Enteric Pathogen Clostridium difficile PCR Ribotype 078 Spores in Food.

Emerging enteric pathogens could have not only more antibiotic resistance or virulence traits; they could also have increased resistance to heat. We quantified the effects of minimum recommended cooking and higher temperatures, individually on a collection of C. difficile isolates and on the survival probability of a mixture of emerging C. difficile strains. While minimum recommended cooking time/temperature combinations (63-71°C) allowed concurrently tested strains to survive, higher subboiling temperatures reproducibly favored the selection of newly emerging C. difficile PCR ribotype 078. Survival ratios for "ribotypes 078" : "other ribotypes" (n = 49 : 45 isolates) from the mid-2000s increased from 1 : 1 and 0.7 : 1 at 85°C (for 5 and 10 minutes, resp.) to 2.3 : 1 and 3 : 1 with heating at 96°C (for 5 and 10 minutes, resp.) indicating an interaction effect between the heating temperature and survival of C. difficile genotypes. In multistrain heating experiments, with PCR ribotypes 027 and 078 from 2004 and reference type strain ATCC 9689 banked in the 1970s, multinomial logistic regression (P < 0.01) revealed PCR ribotype 078 was the most resistant to increasing lethal heat treatments. Thermal processes (during cooking or disinfection) may contribute to the selection of emergent specific virulent strains of C. difficile. Despite growing understanding of the role of cooking on human evolution, little is known about the role of cooking temperatures on the selection and evolution of enteric pathogens, especially spore-forming bacteria.

Soil Conditions That Can Alter Natural Suppression of Escherichia coli O157:H7 in Ohio Specialty Crop Soils.

Food-borne pathogen persistence in soil fundamentally affects the production of safe vegetables and small fruits. Interventions that reduce pathogen survival in soil would have positive impacts on food safety by minimizing preharvest contamination entering the food chain. Laboratory-controlled studies determined the effects of soil pH, moisture content, and soil organic matter (SOM) on the survivability of this pathogen through the creation of single-parameter gradients. Longitudinal field-based studies were conducted in Ohio to quantify the extent to which field soils suppressed Escherichia coli O157:H7 survival. In all experiments, heat-sensitive microorganisms were responsible for the suppression of E. coli O157 in soil regardless of the chemical composition of the soil. In laboratory-based studies, soil pH and moisture content were primary drivers of E. coli O157 survival, with increases in pH after 48 h (P = 0.02) and decreases in moisture content after 48 h (P = 0.007) significantly increasing the log reduction of E. coli O157 numbers. In field-based experiments, E. coli O157 counts from both heated and unheated samples were sensitive to both season (P = 0.004 for heated samples and P = 0.001 for unheated samples) and region (P = 0.002 for heated samples and P = 0.001 for unheated samples). SOM was observed to be a more significant driver of pathogen suppression than the other two factors after 48 h at both planting and harvest (P = 0.002 at planting and P = 0.058 at harvest). This research reinforces the need for both laboratory-controlled experiments and longitudinal field-based experiments to unravel the complex relationships controlling the survival of introduced organisms in soil.

Three-week summer period prevalence of Clostridium difficile in farm animals in a temperate region of the United States (Ohio).

The cross-sectional (period) prevalence of Clostridium difficile in 875 farm animals from 29 commercial operations during the summer of 2008 in Ohio, USA was quantified. Compared to an external referent population of intensively managed race horses (12.7%), intensively managed commercially mature food animals (poultry, cattle, swine; < 0.6%) were infrequent shedders of C. difficile (P < 0.00001) during the warmest weeks of 2008.

Prévalence deClostridium difficilependant une période de trois semaines en été chez les animaux de ferme dans une région tempérée des États-Unis (Ohio). La prévalence par période de Clostridium difficile chez 875 animaux de ferme provenant de 29 exploitations commerciales durant l'été de 2008 en Ohio, aux États-Unis, est quantifiée. Comparativement à la population de référence constituée externe de chevaux de course à gestion intensive (12,7 %), les animaux destinés à l'alimentation et prêts à la commercialisation (volaille, bovins, porcs; < 0,6 %) ont rarement excrété C. difficile (P < 0,00001) durant les semaines les plus chaudes de 2008.(Traduit par Isabelle Vallières).

Molecular epidemiology and public health relevance of Campylobacter isolated from dairy cattle and European starlings in Ohio, USA.

Dairy cattle serve as a potential source for Campylobacter infection in humans. Outbreaks associated with consumption of either Campylobacter contaminated raw milk or contaminated milk after treatment were previously recorded in the United States. Further, starlings have been implicated in the spread of bacterial pathogens among livestock. Here, we determined the prevalence, genotypic, and phenotypic properties of Campylobacter isolated from fecal samples of dairy cattle and starlings found on the same establishment in northeastern Ohio. Campylobacter were detected in 83 (36.6%) and 57 (50.4%) out of 227 dairy and 113 starling fecal samples, respectively. Specifically, 79 C. jejuni, five C. coli, and two other Campylobacter spp. were isolated from dairy feces, while all isolates from starlings (n=57) were C. jejuni. Our results showed that the prevalence of C. jejuni in birds was significantly (p<0.01) higher than that in dairy cattle. The pulsed-field gel electrophoresis analysis showed that C. jejuni were genotypically diverse and host restricted; however, there were several shared genotypes between dairy cattle and starling isolates. Likewise, many shared clonal complexes (CC) between dairy cattle and starlings were observed by multilocus sequence typing (MLST) analysis. As in humans, both in cattle and starlings, the CC 45 and CC 21 were the most frequently represented CCs. As previously reported, CC 177 and CC 682 were restricted to the bird isolates, while CC 42 was restricted to dairy cattle isolates. Further, two new sequence types (STs) were detected in C. jejuni from dairy cattle. Interestingly, cattle and starling C. jejuni showed high resistance to multiple antimicrobials, including ciprofloxacin, erythromycin, and gentamicin. In conclusion, our results highlight starlings as potential reservoirs for C. jejuni, and they may play an important role in the epidemiology of clinically important C. jejuni in dairy population.

Use of multiple-locus variable-number tandem repeat analysis to evaluate Escherichia coli O157 subtype distribution and transmission dynamics following natural exposure on a closed beef feedlot facility.

To better understand the epizootiology of Escherichia coli O157:H7 among cattle, all E. coli O157 isolates recovered on a research feedlot during a single feeding period were characterized by multiple-locus variable-number tandem repeat analysis (MLVA). Three distinct MLVA subtypes (A, B, C), accounting for 24%, 15%, and 64% of total isolates, respectively, were identified. Subtypes A and B were isolated at the initiation of sampling, but their prevalence waned and subtype C, first isolated on the third sampling date, became the predominant subtype on the feedlot. Supershedding events, however, occurred with equal frequency for all three MLVA-types. Using a multilevel logistic regression model, we investigated whether the odds of shedding subtype C relative to subtypes A or B were associated with time, diet, or the presence of a penmate shedding high numbers of subtype C. Only time and exposure to an animal shedding MLVA-type C at 10³ colony-forming units or greater in the pen at the time of sampling were significantly associated with increased shedding of subtype C. High-level shedding of those E. coli O157 subtypes better suited for survival in the environment and/or in the host appear to play a significant role in the development of predominant E. coli O157 subtypes. Supershedding events alone are neither required nor sufficient to drive the epidemiology of specific E. coli O157 subtypes. Additional factors are necessary to direct successful on-farm transmission of E. coli O157.

Shiga toxin (stx) encoding genes in sheep and goats reared in Trinidad and Tobago.

Shiga toxin-producing Escherichia coli (STEC) is estimated to cause over two million cases of human disease annually. Trinidad and Tobago is one of the largest livestock producer and consumer of sheep and goat meat in the Caribbean, however, the potential role of these animals in the epidemiology of STEC infections has not been previously described. To fill this critical gap in knowledge, the prevalence of Shiga toxin genes (stx1 and stx2) shed in the faeces of healthy sheep (n = 204) and goats (n = 105) in Trinidad was investigated. Based on PCR screening, goats had a higher stx prevalence than sheep (46% vs 35%, P = 0.06). Most of the recovered STEC isolates were positive for stx1 only; and only three isolates were positive for the eae gene. None of the recovered isolates belonged to the O157 serogroup. In both species, the prevalence of stx was higher in young animals versus older animals. Sheep reared on deep litter flooring (43%) had a higher prevalence than sheep reared other flooring types, however this was not the same for goats. The presence of cows on the same premise was not an associated predictor for STEC carriage in sheep or goats. This study demonstrates that although sheep and goats in Trinidad are reservoirs for stx-positive E. coli isolates, no fecal samples tested positive for O157 STEC, harbored. Furthermore, it appears that non-O157 stx-positive isolates harbored by these animals do not pose a significant threat to human health.

Identification and Subtyping of Salmonella Isolates Using Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF).

Subtyping of bacterial isolates of the same genus and species is an important tool in epidemiological investigations. A number of phenotypic and genotypic subtyping methods are available; however, most of these methods are labor-intensive and time-consuming and require considerable operator skill and a wealth of reagents. Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF), an alternative to conventional subtyping methods, offers a rapid, reproducible method for bacterial identification with a high sensitivity and specificity and at minimal cost. The purpose of this study was to determine the feasibility of using MALDI-TOF to differentiate between six Salmonella serovars recovered from experimental microcosms inoculated with known strains of Salmonella. Following the establishment of a MALDI-TOF reference library for this project, the identity of 843 Salmonella isolates recovered from these microcosms was assessed using both MALDI-TOF and conventional methods (serotyping/PCR). All 843 isolates were identified as being Salmonella species. Overall, 803/843 (95%) of these isolates were identified similarly using the two different methods. Positive percent agreement at the serovar level ranged from 79 to 100%, and negative percent agreement for all serovars was greater than 98%. Cohen's kappa ranged from 0.85 to 0.98 for the different serovars. This study demonstrates that MALDI-TOF is a viable alternative for the rapid identification and differentiation of Salmonella serovars.

Moist-heat resistance, spore aging, and superdormancy in Clostridium difficile.

Clostridium difficile spores can survive extended heating at 71°C (160°F), a minimum temperature commonly recommended for adequate cooking of meats. To determine the extent to which higher temperatures would be more effective at killing C. difficile, we quantified (D values) the effect of moist heat at 85°C (145°F, for 0 to 30 min) on C. difficile spores and compared it to the effects at 71 and 63°C. Fresh (1-week-old) and aged (≥20-week-old) C. difficile spores from food and food animals were tested in multiple experiments. Heating at 85°C markedly reduced spore recovery in all experiments (5 to 6 log(10) within 15 min of heating; P < 0.001), regardless of spore age. In ground beef, the inhibitory effect of 85°C was also reproducible (P < 0.001), but heating at 96°C reduced 6 log(10) within 1 to 2 min. Mechanistically, optical density and enumeration experiments indicated that 85°C inhibits cell division but not germination, but the inhibitory effect was reversible in some spores. Heating at 63°C reduced counts for fresh spores (1 log(10), 30 min; P < 0.04) but increased counts of 20-week-old spores by 30% (15 min; P < 0.02), indicating that sublethal heat treatment reactivates superdormant spores. Superdormancy is an increasingly recognized characteristic in Bacillus spp., and it is likely to occur in C. difficile as spores age. The potential for reactivation of (super)dormant spores with sublethal temperatures may be a food safety concern, but it also has potential diagnostic value. Ensuring that food is heated to >85°C would be a simple and important intervention to reduce the risk of inadvertent ingestion of C. difficile spores.

Transient fecal shedding and limited animal-to-animal transmission of Clostridium difficile by naturally infected finishing feedlot cattle.

To longitudinally assess fecal shedding and animal-to-animal transmission of Clostridium difficile among finishing feedlot cattle as a risk for beef carcass contamination, we tested 186 ± 12 steers (mean ± standard deviation; 1,369 samples) in an experimental feedlot facility during the finishing period and at harvest. Clostridium difficile was isolated from 12.9% of steers on arrival (24/186; 0 to 33% among five suppliers). Shedding decreased to undetectable levels a week later (0%; P < 0.001), and remained low (< 3.6%) until immediately prior to shipment for harvest (1.2%). Antimicrobial use did not increase fecal shedding, despite treatment of 53% of animals for signs of respiratory disease. Animals shedding C. difficile on arrival, however, had 4.6 times higher odds of receiving antimicrobials for respiratory signs than nonshedders (95% confidence interval for the odds ratio, 1.4 to 14.8; P = 0.01). Neither the toxin genes nor toxin A or B was detected in most (39/42) isolates based on two complementary multiplex PCRs and enzyme-linked immunosorbent assay testing, respectively. Two linezolid- and clindamycin-resistant PCR ribotype 078 (tcdA+/tcdB+/cdtB+/39-bp-type deletion in tcdC) isolates were identified from two steers (at arrival and week 20), but these ribotypes did not become endemic. The other toxigenic isolate (tcdA+/tcdB+/cdtB+/classic tcdC; PCR ribotype 078-like) was identified in the cecum of one steer at harvest. Spatio-temporal analysis indicated transient shedding with no evidence of animal-to-animal transmission. The association between C. difficile shedding upon arrival and the subsequent need for antimicrobials for respiratory disease might indicate common predisposing factors. The isolation of toxigenic C. difficile from bovine intestines at harvest highlights the potential for food contamination in meat processing plants.

General suppression of Escherichia coli O157:H7 in sand-based dairy livestock bedding.

Sand bedding material is frequently used in dairy operations to reduce the occurrence of mastitis and enhance cow comfort. One objective of this work was to determine if sand-based bedding also supported the microbiologically based suppression of an introduced bacterial pathogen. Bedding samples were collected in summer, fall, and winter from various locations within a dairy operation and tested for their ability to suppress introduced populations of Escherichia coli O157:H7. All sources of bedding displayed a heat-sensitive suppressiveness to the pathogen. Differences in suppressiveness were also noted between different samples at room temperature. At just 1 day postinoculation (dpi), the recycled sand bedding catalyzed up to a 1,000-fold reduction in E. coli counts, typically 10-fold greater than the reduction achieved with other substrates, depending on the sampling date. All bedding substrates were able to reduce E. coli populations by over 10,000-fold within 7 to 15 dpi, regardless of sampling date. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to identify bacterial populations potentially associated with the noted suppression of E. coli O157:H7 in sand bedding. Eleven terminal restriction fragments (TRFs) were overrepresented in paired comparisons of suppressive and nonsuppressive specimens at multiple sampling points, indicating that they may represent environmentally stable populations of pathogen-suppressing bacteria. Cloning and sequencing of these TRFs indicated that they represent a diverse subset of bacteria, belonging to the Cytophaga-Flexibacter-Bacteroidetes, Gammaproteobacteria, and Firmicutes, only a few of which have previously been identified in livestock manure. Such data indicate that microbial suppression may be harnessed to develop new options for mitigating the risk and dispersal of zoonotic bacterial pathogens on dairy farms.

Assessment of diagnostic tools for identifying cattle shedding and super-shedding Escherichia coli O157:H7 in a longitudinal study of naturally infected feedlot steers in Ohio.

The objectives of this study were to compare the performance of different diagnostic protocols (rectoanal mucosal swabs and immunomagnetic separation [RAMS-IMS], fecal samples and IMS [fecal-IMS], and direct plating) to determine the prevalence of Escherichia coli O157:H7 and to evaluate the pattern of E. coli O157:H7 shedding and super-shedding (defined as having a direct plating count equal to or >10(4) colony forming units of E. coli O157:H7 per gram of feces) in a longitudinal study of naturally infected feedlot steers. RAMS and fecal grab samples were obtained at 14-day intervals from 168 Angus-cross beef steers over a period of 22 weeks. Fecal samples were assessed by direct plating and IMS, whereas RAMS were tested only by enrichment followed by IMS to recover E. coli O157:H7. The period prevalence for shedding was high (62%) among feedlot steers and super-shedding was higher (23%) than anticipated. Although direct plating was the least sensitive method to detect E. coli O157:H7-positive samples, over 20% of high bacterial load samples were not detected by RAMS-IMS and/or fecal-IMS. The sensitivity of RAMS-IMS, fecal-IMS, and direct plating protocols was estimated using simple and multilevel mixed-effects logistic regression models, in which the dependent variable was the dichotomous results of each test and gold standard (i.e., parallel interpretation of the three protocols)-positive individuals were included as an independent variable along with other factors such as dietary supplements, time of sampling, and being exposed to a super-shedding pen-mate. The associations between these factors and the sensitivity of the diagnostic protocols were not statistically significant. In conclusion, differences in the reported impact of diet and probiotics on the shedding of E. coli O157:H7 in previous studies using RAMS-IMS or fecal-IMS were unlikely due to their impact on test performance.

FAO/WHO Joint Expert Meeting on Microbiological Risk Assessment (JEMRA): Twenty Years of International Microbiological Risk Assessment.

Since the late 1990s, the Food and Agriculture Organization (FAO) of the United Nations and the World Health Organization (WHO) has convened expert meetings and consultations to address the microbiological risk assessment (MRA). These meetings are held to provide scientific advice in response to requests for from Codex Alimentarius, the international food standard-setting body. Individuals participate in the FAO/WHO joint expert meetings on the microbiological risk assessment (JEMRA) in their personal capacity, as technical experts, yet bring diverse regional and national perspectives that contribute to practical applications, particularly for low- and middle-income countries (LMICs). Over 370 experts from around the globe have contributed to the meeting outcomes that have been published in nearly 40 monographs in the FAO/WHO microbial risk assessment (MRA) series, addressing particular food commodities with microbial hazard(s) combinations or a methodological aspect of microbial risk assessment. FAO/WHO MRA series inform Codex decision-making for the development of international standards for safe food and faire trade in food products; are consulted by risk managers such as food safety authorities and food business operators to make science-based decisions; and are used by academics to advance food safety research and educate the next generation of food safety professionals.

Temporal Trends in Antimicrobial Resistance of Fecal Escherichia coli from Deer.

The changing epidemiologic role of wildlife as reservoirs of antimicrobial-resistant bacteria (ARB) is poorly understood. In this study, we characterize the phenotypic resistance of commensal Escherichia coli from fecal samples of 879 individual white-tailed (Odocoileus virginianus; WTD) over a ten-year period and analyze resistance patterns. Our results show commensal E. coli from WTD had significant linear increases in reduced susceptibility to 5 of 12 antimicrobials, including broad-spectrum cephalosporins and fluoroquinolones, from 2006 to 2016. In addition, the relative frequency distribution of minimal inhibitory concentrations of two additional antimicrobials shifted towards higher values from across the study period. The prevalence of multidrug-resistant commensal E. coli increased over the study period with a prevalence of 0%, 2.2%, and 3.7% in 2006, 2012, and 2016, respectively. WTD may be persistently and increasingly exposed to antibiotics or their residues, ARB, and/or antimicrobial resistance genes via contaminated environments like surface water receiving treated wastewater effluent.

Low prevalence of Escherichia coli O157:H7 in horses in Ohio, USA.

Manure from draft animals deposited in fields during vegetable and fruit production may serve as a potential source of preharvest pathogen contamination of foods. To better quantify this risk, we determined the prevalence of Escherichia coli O157:H7 in horses. Between June and September 2009, freshly voided fecal samples were collected from horses stabled on 242 separate premises in Ohio, USA. Overall, the prevalence of E. coli O157:H7 was 1 of 242 (0.4% prevalence, 95% confidence interval [CI] = 0.01 to 2.28). E. coli O157:H7 was recovered from none of the 107 equine fecal samples (0% prevalence, 95% CI = 0.00 to 3.39) that originated from locations without ruminant presence, and only 1 of the 135 horse fecal samples (0.7% prevalence, 95% CI = 0.02 to 4.06) from sites where ruminants were also present. The lone positive sample was collected from a horse that was costabled with a goat. Subsequent sampling at that location identified indistinguishable subtypes of E. coli O157:H7 present in the cohoused goat, in the environment, insects, sheep, and other goats housed in an adjacent field. E. coli O157:H7 was not isolated from the five subsequent samples from this horse. These data indicate that E. coli O157:H7 carriage by horses is an uncommon event.

Enteric bacterial pathogens with zoonotic potential isolated from farm-raised deer.

The raising of captive white-tailed deer (Odocoileus virginianus) is a growing agricultural industry in Ohio as it is in several other areas of the United States and around the world. Pooled fecal samples were collected from 30 white-tailed deer confinement facilities. Samples were cultured for five enteric bacterial pathogens. Premise prevalence rates were as follows: Escherichia coli O157, 3.3%; Listeria monocytogenes, 3.3%; Salmonella enterica, 0%; Yersinia enterocolitica, 30%; and Clostridium difficile, 36.7%. The ail virulence gene could not be amplified from any of the Y. enterocolitica isolates recovered. Toxigenic strains of C. difficile polymerase chain reaction ribotype 078, an emerging C. difficile genotype of humans and food animals, were recovered from 4 of 11 (36.4%) C. difficile-positive deer farms. Venison from farm-raised deer might become contaminated with foodborne pathogens, deer farmers may have occupational exposure to these zoonotic agents, and farm-raised deer could be a reservoir from which the environment and other livestock may become contaminated with a number of potentially zoonotic bacteria.

Differences in Listeria monocytogenes contamination of rural Ohio residences with and without livestock.

To determine the contribution of on-site livestock to the environmental contamination of rural households with Listeria monocytogenes, a total of 1779 environmental and food samples were collected from 26 ruminant-farm households and 26 rural households in Ohio. L. monocytogenes isolates were identified and differentiated using sequence comparisons of the intragenic regions of inlB and inlC. L. monocytogenes was isolated from shoes, 9.6% (20/208); utility gloves, 5.4% (6/111); kitchen sinks, 1.5% (3/204); washing machines, 0.96% (2/204); food, 1.11% (7/631); and animal feces, 8.7% (9/104), over the course of four household visits at monthly intervals. Notably, L. monocytogenes-contaminated shoes were identified more frequently from ruminant farmhouses than from rural households that did not raise ruminants on site (odds ratio = 4.8). L. monocytogenes isolated from animal feces was indistinguishable from strains recovered from shoes and gloves stored in several homes. Our results highlight the potential of the rural household environment as source of L. monocytogenes exposure.

A randomized controlled trial to assess the impact of dietary energy sources, feed supplements, and the presence of super-shedders on the detection of Escherichia coli O157:H7 in feedlot cattle using different diagnostic procedures.

Alteration of the gastro-intestinal tract through manipulation of cattle diets has been proposed as a preharvest control measure to reduce fecal shedding of Escherichia coli O157:H7. The objective of this study was to examine the effects of the energy source's moisture content (high moisture corn and dry whole-shelled corn), two natural feed supplements (Saccaromyces cerevisiae boulardii CNCM 1079-Levucell and Aspergillus oryzae-Amaferm), and two levels of vitamin A (2200 IU/kg and no supplementation) on the fecal excretion of E. coli O157:H7 in naturally infected cattle. One hundred sixty-eight Angus-cross beef steers were randomly allocated to 24 pens, and each pen was assigned 1 of 12 dietary treatments in a randomized complete block design. E. coli O157:H7 was detected by rectoanal mucosal swab (RAMS) and fecal grab samples using immunomagnetic separation (IMS) and standard microbiological techniques. On the basis of multivariable multilevel logistic regression models, we found a statistically significant (p < 0.05) increase in the prevalence of E. coli O157:H7 in animals fed dry whole-shelled corn in models based on fecal-IMS, and this effect was increased if a super-shedding animal (shedding > 10(4) colony forming units of E. coli O157:H7 per gram of feces) was present in the pen at the time of testing relative to animals fed high moisture corn and nonexposed to super-shedders. However, in similar models based on RAMS-IMS testing, the effect of corn type on the prevalence of E. coli O157:H7 varied with the type of feed additive used. Being exposed to a super-shedding pen-mate also increased the odds of being positive to E. coli O157:H7 in the RAMS-IMS models. These models demonstrate that the impact of different supplements may vary with the diagnostic test used, and that further research into the biological significance of differences between RAMS- and fecal-IMS test results is warranted.

Bird-livestock interactions associated with increased cattle fecal shedding of ciprofloxacin-resistant Escherichia coli within feedlots in the United States.

This research study was conducted to determine if bird depredation in feedlots is associated with the prevalence of ciprofloxacin-resistant Escherichia coli in cattle and to determine if removal of invasive bird species could be an effective management strategy to help reduce ciprofloxacin-resistant E. coli in cattle within the United States. European starlings (Sturnus vulgaris) were collected from feedlots within multiple geographic regions within the United States and European starlings within all regions tested positive for ciprofloxacin-resistant E. coli, but prevalence differed by region. Total number of birds on feedlots were positively associated with increased cattle fecal shedding of ciprofloxacin-resistant E. coli. Targeted control of invasive European starlings reduced bird numbers on feedlots by 70.4%, but decreasing populations of European starlings was not associated with corresponding reductions in bovine fecal prevalence of ciprofloxacin-resistant E. coli. These data provide evidence for the role of wild bird depredation in feedlots contributing to fecal shedding of ciprofloxacin-resistant E. coli, but a single month of European starling control in feedlots was not sufficient to impact the fecal carriage of this organism in cattle.