Antibimicrobial Susceptibility of Trimethoprim-Sulfamethoxazole and 3rd-Generation Cephalosporin-Resistant Escherichia coli Isolates Enumerated Longitudinally from Feedlot Arrival to Harvest in High-Risk Beef Cattle Administered Common Metaphylactic Antimicrobials.

Multidrug resistant (MDR) Escherichia coli threaten the preservation of antimicrobials to treat infections in humans and livestock. Thus, it is important to understand where antimicrobial-resistant E. coli persist and factors that contribute to its their development. Crossbred cattle (n = 249; body weight = 244 kg ±25 kg standard deviation) were blocked by arrival date and assigned metaphylactic antimicrobial treatments of sterile saline control, tulathromycin (TUL), ceftiofur, or florfenicol at random. Trimethoprim-sulfamethoxazole (COTR) and third-generation cephalosporin (CTXR)-resistant E. coli were isolated from fecal samples on days 0, 28, 56, 112, 182, and study END (day 252 for block 1 and day 242 for block 2). Then, susceptibility testing was conducted on all confirmed isolates. MDR was detected in both COTR and CTXR E. coli isolates. In COTR isolates, the number of antimicrobials each isolate was resistant to and the minimum inhibitory concentration (MIC) for amoxicillin-clavulanic acid, ceftriaxone, and gentamicin was greatest on day 28 compared with all other days (p ≤ 0.04). Similarly, chloramphenicol MIC was greater on day 28 than on day 0 (p < 0.01). Overall, sulfisoxazole MIC was less for TUL than all other treatments (p ≤ 0.02), and trimethoprim-sulfamethoxazole MIC was greater for TUL than all other treatments (p ≤ 0.03). Finally, there was no effect of treatment, day, or treatment × day for tetracycline or meropenem MIC (p ≥ 0.07). In CTXR isolates, there was an effect of day for all antimicrobials tested except ampicillin and meropenem (p ≤ 0.06). In conclusion, administering a metaphylactic antimicrobial at feedlot arrival did influence the susceptibility of COTR and CTXR E. coli. However, MDR E. coli are widely distributed, and the MIC for most antimicrobials was not different from the initial value upon completion of the feeding period.

Metaphylactic antimicrobial effects on occurrences of antimicrobial resistance in Salmonella enterica, Escherichia coli and Enterococcus spp. measured longitudinally from feedlot arrival to harvest in high-risk beef cattle.

AIMS: Our objective was to determine how injectable antimicrobials affected populations of Salmonella enterica, Escherichia coli and Enterococcus spp. in feedlot cattle. METHODS AND RESULTS: Two arrival date blocks of high-risk crossbred beef cattle (n = 249; mean BW = 244 kg) were randomly assigned one of four antimicrobial treatments administered on day 0: sterile saline control (CON), tulathromycin (TUL), ceftiofur (CEF) or florfenicol (FLR). Faecal samples were collected on days 0, 28, 56, 112, 182 and study end (day 252 for block 1 and day 242 for block 2). Hide swabs and subiliac lymph nodes were collected the day before and the day of harvest. Samples were cultured for antimicrobial-resistant Salmonella, Escherichia coli and Enterococcus spp. The effect of treatment varied by day across all targeted bacterial populations (p ≤ 0.01) except total E. coli. Total E. coli counts were greatest on days 112, 182 and study end (p ≤ 0.01). Tulathromycin resulted in greater counts and prevalence of Salmonella from faeces than CON at study end (p ≤ 0.01). Tulathromycin and CEF yielded greater Salmonella hide prevalence and greater counts of 128ERYR E. coli at study end than CON (p ≤ 0.01). No faecal Salmonella resistant to tetracyclines or third-generation cephalosporins were detected. Ceftiofur was associated with greater counts of 8ERYR Enterococcus spp. at study end (p ≤ 0.03). By the day before harvest, antimicrobial use did not increase prevalence or counts for all other bacterial populations compared with CON (p ≥ 0.13). CONCLUSIONS: Antimicrobial resistance (AMR) in feedlot cattle is not caused solely by using a metaphylactic antimicrobial on arrival, but more likely a multitude of environmental and management factors.

Management-intensive grazing impacts on total Escherichia coli, E. coli O157:H7, and antibiotic resistance genes in a riparian stream.

The impacts of management-intensive grazing (MIG) of cattle on concentrations of total Escherichia coli, total suspended solids (TSS), and nitrate-nitrite nitrogen (NO3 + NO2-N), and occurrence of E. coli O157:H7 and selected antibiotic resistance genes (ARGs) in stream water and/or sediments were evaluated. Cattle were grazed for two-week periods in May in each of three years. Overall, grazing increased total E. coli in downstream water by 0.89 log10 MPN/100 mL (p < 0.0001), and downstream total E. coli concentrations were higher than upstream over all sampling intervals. Downstream TSS levels also increased (p ≤ 0.0294) during grazing. In contrast, there was a main effect of treatment for downstream NO3 + NO2-N to be lower than upstream (3.59 versus 3.70 mg/L; p = 0.0323). Overwintering mallard ducks increased total E. coli and TSS concentrations in January and February (p < 0.05). For precipitation events during the 24 h before sampling, each increase of 1.00 cm of rainfall increased total E. coli by 0.49 log10 MPN/100 mL (p = 0.0005). In contrast, there was no association of previous 24 h precipitation volume on TSS (p = 0.1540), and there was a negative linear effect on NO3 + NO2-N (p = 0.0002). E. coli O157:H7 prevalence was low, but the pathogen was detected downstream up to 2½ months after grazing. Examination of ARGs sul1, ermB, blactx-m-32, and intI1 identified the need for additional research to understand the impact of grazing on the ecology of these resistance determinants in pasture-based cattle production. While E. coli remained higher in downstream water compared to upstream, MIG may reduce the magnitude of the downstream E. coli concentrations. Likewise, the MIG strategy may prevent large increases in TSS and NO3 + NO2-N concentrations during heavy rain events. Results indicate that MIG can limit the negative effects of cattle grazing on stream water quality.

Measuring the occurrence of antibiotics in surface water adjacent to cattle grazing areas using passive samplers.

A wide variety of antibiotics and other pharmaceuticals are used in livestock production systems and residues passed to the environment, often unmetabolized, after use and excretion. Antibiotic residues may be transported from manure-treated soils via runoff and are also capable of reaching surface and groundwater systems through a variety of pathways. The occurrence and persistence of antibiotics in the environment is a concern due to the potential for ecological effects and proliferation of environmental antibiotic resistance in pathogenic organisms. In the present study, the occurrence and seasonal variation of 24 commonly-used veterinary antibiotics was evaluated in surface water adjacent to several livestock production systems using Polar Organic Chemical Integrative Samplers (POCIS). Uptake rates for all compounds, nine of which have not been previously reported, were measured in the laboratory to permit estimation of changes in the time-weighted average (TWA) antibiotic concentrations during exposure. The antibiotics detected in POCIS extracts included sulfadimethoxine, sulfamethoxazole, trimethoprim, sulfamerazine, sulfadiazine, lincomycin, erythromycin, erythromycin anhydro- and monensin. The maximum TWA concentration belonged to sulfadiazine (25 ng/L) in the August-September sampling period and coincided with the highest number of precipitation events. With the exception of monensin that showed an increase in concentration over the stream path, none of the detected antibiotics were prescribed to livestock at the facility. The detection of antibiotics not prescribed by the facility may be attributable to the environmental persistence of previously used antibiotics, transfer by wind from other nearby livestock production sites or industrial uses, and/or the natural production of some antibiotics.

Predicting Escherichia coli loads in cascading dams with machine learning: An integration of hydrometeorology, animal density and grazing pattern.

Accurate prediction of Escherichia coli contamination in surface waters is challenging due to considerable uncertainty in the physical, chemical and biological variables that control E. coli occurrence and sources in surface waters. This study proposes a novel approach by integrating hydro-climatic variables as well as animal density and grazing pattern in the feature selection modeling phase to increase E. coli prediction accuracy for two cascading dams at the US Meat Animal Research Center (USMARC), Nebraska. Predictive models were developed using regression techniques and an artificial neural network (ANN). Two adaptive neuro-fuzzy inference system (ANFIS) structures including subtractive clustering and fuzzy c-means (FCM) clustering were also used to develop models for predicting E. coli. The performances of the predictive models were evaluated and compared using root mean squared log error (RMSLE). Cross-validation and model performance results indicated that although the majority of models predicted E. coli accurately, ANFIS models resulted in fewer errors compared to the other models. The ANFIS models have the potential to be used to predict E. coli concentration for intervention plans and monitoring programs for cascading dams, and to implement effective best management practices for grazing and irrigation during the growing season.

Escherichia coli concentrations in waters of a reservoir system impacted by cattle and migratory waterfowl.

Recent pathogenic Escherichia coli contamination of fresh vegetables that originated from irrigation water has increased awareness and importance of identifying sources of E. coli entering agroecosystems. However, inadequate methods for accurately predicting E. coli occurrence and sources in waterways continue to limit the identification of appropriate and effective prevention and treatment practices. Therefore, the primary objectives of this study were to: (1) Determine the concentration of E. coli during storm events in a hydrologic controlled stream situated in a livestock research operation that is located in the Central Flyway for avian migration in the United States. Great Plains; and (2) Identify trends between E. coli concentrations, grazing rotations, and avian migration patterns. The study sampled five rainfall events (three summer and two fall) to measure E. coli concentrations throughout storm events. A combination of cattle density and waterfowl migration patterns were found to significantly impact E. coli concentrations in the stream. Cattle density had a significant impact during the summer season (p < .0001), while waterfowl density had a significant impact on E. coli concentrations during the fall (p = .0422). The downstream reservoir had exceedance probabilities above the Environmental Protection Agency freshwater criteria > 85% of the growing season following rainfall events. Based on these findings, implementation of best management practices for reducing E. coli concentrations during the growing season and testing of irrigation water prior to application are recommended.

Occurrence of Escherichia coli O157:H7 in Pest Flies Captured in Leafy Greens Plots Grown Near a Beef Cattle Feedlot.

Leafy greens are leading vehicles for Escherichia coli O157:H7 foodborne illness. Pest flies can harbor this pathogen and may disseminate it to produce. We determined the occurrence of E. coli O157:H7-positive flies in leafy greens planted up to 180 m from a cattle feedlot and assessed their relative risk to transmit this pathogen to leafy greens. The primary fly groups captured on sticky traps at the feedlot and leafy greens plots included house flies (Musca domestica L.), face flies (Musca autumnalis L.), stable flies (Stomoxys calcitrans L.), flesh flies (family Sarcophagidae), and blow flies (family Calliphoridae). E. coli O157:H7 carriage rates of house, face, flesh, and blow flies were similar (P > 0.05), ranging from 22.3 to 29.0 flies per 1,000 flies. In contrast, the carriage rate of stable flies was lower at 1.1 flies per 1,000 flies (P < 0.05). Differences in carriage rates are likely due to the uses of fresh bovine feces and manure by these different pest fly groups. E. coli O157:H7 carriage rates of total flies did not differ (P > 0.05) by distance (ranging from 0 to 180 m) from the feedlot. Most fly isolates were the same predominant pulsed-field gel electrophoresis types found in feedlot surface manure and leafy greens, suggesting a possible role for flies in transmitting E. coli O157:H7 to the leafy greens. However, further research is needed to clarify this role and to determine set-back distances between cattle production facilities and produce crops that will reduce the risk for pathogen contamination by challenging mechanisms like flies.

Energy balance affects pulsatile secretion of luteinizing hormone from the adenohypophesis and expression of neurokinin B in the hypothalamus of ovariectomized gilts.

The pubertal transition of gonadotropin secretion in pigs is metabolically gated. Kisspeptin (KISS1) and neurokinin B (NKB) are coexpressed in neurons within the arcuate nucleus of the hypothalamus (ARC) and are thought to play an important role in the integration of nutrition and metabolic state with the reproductive neuroendocrine axis. The hypothesis that circulating concentrations of luteinizing hormone (LH) and expression of KISS1 and tachykinin 3(TAC3, encodes NKB) in the ARC of female pigs are reduced with negative energy balance was tested using ovariectomized, prepubertal gilts fed to either gain or lose body weight. Restricted feeding of ovariectomized gilts caused a rapid and sustained metabolic response characterized by reduced concentrations of plasma urea nitrogen, insulin, leptin, and insulin-like growth factor-1 and elevated concentrations of free fatty acids. The secretory pattern of LH shifted from one of low amplitude to one of high amplitude, which caused overall circulating concentrations of LH to be greater in restricted gilts. Nutrient-restricted gilts had greater expression of follicle-stimulating hormone and gonadotropin-releasing hormone receptor, but not LH in the anterior pituitary gland. Expression of KISS1 in the ARC was not affected by dietary treatment, but expression of TAC3 was greater in restricted gilts. These data are consistent with the idea that hypothalamic expression of KISS1 is correlated with the number of LH pulse in pig, and further indicate that amplitude of LH pulses may be regulated by NKB in the gilt.

Rumen bacterial community structure impacts feed efficiency in beef cattle.

The importance of the rumen microbiota on nutrient cycling to the animal is well recognized; however, our understanding of the influence of the rumen microbiome composition on feed efficiency is limited. The rumen microbiomes of two large animal cohorts (125 heifers and 122 steers) were characterized to identify specific bacterial members (operational taxonomic units [OTUs]) associated with feed efficiency traits (ADFI, ADG, and G:F) in beef cattle. The heifer and steer cohorts were fed a forage-based diet and a concentrate-based diet, respectively. A rumen sample was obtained from each animal via esophageal tubing and bacterial community composition was determined through 16S rRNA gene sequencing of the V4 region. Based on a regression approach that used individual performance measures, animals were classified into divergent feed efficiency groups. Within cohort, an extreme set of 16 animals from these divergent groups was selected as a discovery population to identify differentially abundant OTUs across the rumen bacterial communities. The remaining samples from each cohort were selected to perform forward stepwise regressions using the differentially abundant OTUs as explanatory variables to distinguish predictive OTUs for the feed efficiency traits and to quantify the OTUs collective impact on feed efficiency phenotypes. OTUs belonging to the families Prevotellaceae and Victivallaceae were present across models for heifers, whereas OTUs belonging to the families Prevotellaceae and Lachnospiraceae were present across models for steers. Within the heifer cohort, models explained 19.3%, 25.3%, and 19.8% of the variation for ADFI, ADG, and G:F, respectively. Within the steer cohort, models explained 27.7%, 32.5%, and 26.9% of the variation for ADFI, ADG, and G:F, respectively. Overall, this study suggests a substantial role of the rumen microbiome on feed efficiency responses.

Odorous Volatile Organic Compounds, Escherichia coli, and Nutrient Concentrations when Kiln-Dried Pine Chips and Corn Stover Bedding Are Used in Beef Bedded Manure Packs.

Pine ( spp.) bedding has been shown to lower the concentration of odorous volatile organic compounds (VOCs) and pathogenic bacteria compared with corn ( L.) stover bedding, but availability and cost limit the use of pine bedding in cattle confinement facilities. The objectives of this study were to determine if the addition of pine wood chips to laboratory-scaled bedded packs containing corn stover (i) reduced odorous VOC emissions; (ii) reduced total ; and (iii) changed the nutrient composition of the resulting manure-bedded packs. Bedding treatments included 0, 10, 20, 30, 40, 60, 80, and 100% pine chips, with the balance being corn stover. Four bedded packs for each mixture were maintained for 42 d ( = 4 observations per bedding material). The production of total sulfur compounds increased significantly when 100% pine chips were used (44.72 ng L) compared with bedding mixture containing corn stover (18.0-24.56 ng L). The carbon-to-nitrogen ratio exceeded the ideal ratio of 24:1 for the optimum activity of soil microorganisms when ≥60% pine chips (25.3-27.5 ng L) were included in the mixture. The use of 100% pine chips as bedding increased sulfide concentration in the facility 1.8 to 2.4 times over the use of corn stover bedding. was not influenced by the addition of pine chips to the corn stover bedding material but did decrease as the bedded pack aged. Bedding material mixtures containing 30 to 60% pine and 40 to 70% corn stover may be the ideal combination to mitigate odors from livestock facilities using deep bedded systems.

Evaluation of Commercial ß-Agonists, Dietary Protein, and Shade on Fecal Shedding of Escherichia coli O157:H7 from Feedlot Cattle.

Escherichia coli O157:H7 is a foodborne pathogen commonly associated with cattle feces. Diet, including dietary supplements such as ß-agonists, may impact fecal shedding of this pathogen. A series of three experiments were conducted to determine if the ß-agonists ractopamine hydrochloride (RAC) or zilpaterol hydrochloride (ZH) would impact the level or prevalence of fecal E. coli O157:H7 shedding. In Experiment 1, dietary RAC did not impact fecal shedding of E. coli O157:H7 based on the level or prevalence, but the addition of dietary soybean meal (SBM) in the study did reduce E. coli O157:H7 shedding. In Experiments 2 and 3, dietary ZH did not affect fecal E. coli O157:H7 shedding as determined by enumeration or prevalence, but in Experiment 2 the addition of 30% (dry matter basis) wet distillers grains with solubles (WDGS) in the diet tended to increase E. coli O157:H7 shedding. Shade is a potential management tool to reduce heat stress in cattle, and in Experiment 3 the presence of shade over the feedlot pens did not affect E. coli O157:H7 shedding. The use of ß-agonists in cattle diets did not significantly affect fecal shedding of E. coli O157:H7, and in particular the percentage of animals shedding enumerable levels of the pathogen did not change, indicating that there was not a change in colonization. As has been reported previously and indicated again in this study, the use of WDGS in the diet may increase E. coli O157:H7 shedding. In contrast, the addition of SBM to cattle diets, to increase the dietary crude protein, appeared to reduce E. coli O157:H7 shedding, but this potential dietary intervention needs to be confirmed with additional research.

Persistence of Escherichia coli O157:H7 and Total Escherichia coli in Feces and Feedlot Surface Manure from Cattle Fed Diets with and without Corn or Sorghum Wet Distillers Grains with Solubles.

Feeding corn wet distillers grains with solubles (WDGS) to cattle can increase the load of Escherichia coli O157:H7 in feces and on hides, but the mechanisms are not fully understood. The objective of these experiments was to examine a role for the persistence of E. coli O157:H7 in the feces and feedlot pen surfaces of cattle fed WDGS. In the first study, feces from steers fed 0, 20, 40, or 60% corn WDGS were inoculated with E. coli O157:H7. The E. coli O157:H7 numbers in feces from cattle fed 0% corn WDGS rapidly decreased (P < 0.05), from 6.28 to 2.48 log CFU/g of feces by day 14. In contrast, the E. coli O157:H7 numbers in feces from cattle fed 20, 40, and 60% corn WDGS were 4.21, 5.59, and 6.13 log CFU/g of feces, respectively, on day 14. A second study evaluated the survival of E. coli O157:H7 in feces from cattle fed 0 and 40% corn WDGS. Feces were collected before and 28 days after the dietary corn was switched from high-moisture corn to dry-rolled corn. Within dietary corn source, the pathogen persisted at higher concentrations (P < 0.05) in 40% corn WDGS feces at day 7 than in 0% WDGS. For 40% corn WDGS feces, E. coli O157:H7 persisted at higher concentrations (P < 0.05) at day 7 in feces from cattle fed high-moisture corn (5.36 log CFU/g) than from those fed dry-rolled corn (4.27 log CFU/g). The percentage of WDGS had no effect on the E. coli O157:H7 counts in feces from cattle fed steam-flaked corn-based diets containing 0, 15, and 30% sorghum WDGS. Greater persistence of E. coli O157:H7 on the pen surfaces of animals fed corn WDGS was not demonstrated, although these pens had a higher prevalence of the pathogen in the feedlot surface manure after the cattle were removed. Both or either the greater persistence and higher numbers of E. coli O157:H7 in the environment of cattle fed WDGS may play a part in the increased prevalence of E. coli O157:H7 in cattle by increasing the transmission risk.

Reducing Foodborne Pathogen Persistence and Transmission in Animal Production Environments: Challenges and Opportunities.

Preharvest strategies to reduce zoonotic pathogens in food animals are important components of the farm-to-table food safety continuum. The problem is complex; there are multiple pathogens of concern, multiple animal species under different production and management systems, and a variety of sources of pathogens, including other livestock and domestic animals, wild animals and birds, insects, water, and feed. Preharvest food safety research has identified a number of intervention strategies, including probiotics, direct-fed microbials, competitive exclusion cultures, vaccines, and bacteriophages, in addition to factors that can impact pathogens on-farm, such as seasonality, production systems, diet, and dietary additives. Moreover, this work has revealed both challenges and opportunities for reducing pathogens in food animals. Animals that shed high levels of pathogens and predominant pathogen strains that exhibit long-term persistence appear to play significant roles in maintaining the prevalence of pathogens in animals and their production environment. Continued investigation and advancements in sequencing and other technologies are expected to reveal the mechanisms that result in super-shedding and persistence, in addition to increasing the prospects for selection of pathogen-resistant food animals and understanding of the microbial ecology of the gastrointestinal tract with regard to zoonotic pathogen colonization. It is likely that this continued research will reveal other challenges, which may further indicate potential targets or critical control points for pathogen reduction in livestock. Additional benefits of the preharvest reduction of pathogens in food animals are the reduction of produce, water, and environmental contamination, and thereby lower risk for human illnesses linked to these sources.

Distillers By-Product Cattle Diets Enhance Reduced Sulfur Gas Fluxes from Feedlot Soils and Manures.

Total reduced sulfur (TRS) emissions from animal feeding operations are a concern with increased feeding of high-sulfur distillers by-products. Three feeding trials were conducted to evaluate feeding wet distillers grain plus solubles (WDGS) on TRS fluxes. Fresh manure was collected three times during Feeding Trial 1 from cattle fed 0, 20, 40, and 60% WDGS. Fluxes of TRS from 40 and 60% WDGS manures were 3- to 13-fold greater than the 0 and 20% WDGS manures during the first two periods. In the final period, TRS flux from 60% WDGS was 5- to 22-fold greater than other WDGS manures. During Feeding Trial 2, 0 and 40% WDGS diets on four dates were compared in feedlot-scale pens. On two dates, fluxes from mixed manure and soil near the feed bunk were 3.5-fold greater from 40% WDGS pens. After removing animals, soil TRS flux decreased 82% over 19 d but remained 50% greater in 40% WDGS pens, principally from the wetter pen edges (1.9-fold greater than the drier central mound). During two cycles of cattle production in Feeding Trial 3, TRS soil fluxes were 0.3- to 4-fold greater over six dates for pens feeding WDGS compared with dry-rolled corn diet and principally from wetter pen edges. Soil TRS flux correlated with %WDGS, total N, total P, manure pack temperature, and surface temperature. Consistent results among these three trials indicate that TRS fluxes increase by two- to fivefold when cattle were fed greater levels of WDGS, but specific manure management practices may help control TRS fluxes.

Ammonia, Total Reduced Sulfides, and Greenhouse Gases of Pine Chip and Corn Stover Bedding Packs.

Bedding materials may affect air quality in livestock facilities. Our objective in this study was to compare headspace concentrations of ammonia (NH), total reduced sulfides (TRS), carbon dioxide (CO), methane (CH), and nitrous oxide (NO) when pine wood chips ( spp.) and corn stover ( L.) were mixed in various ratios (0, 10, 20, 30, 40, 60, 80, and 100% pine chips) and used as bedding with manure. Air samples were collected from the headspace of laboratory-scaled bedded manure packs weekly for 42 d. Ammonia concentrations were highest for bedded packs containing 0, 10, and 20% pine chips (equivalent to 501.7, 502.3, and 502.3 mg m, respectively) in the bedding mixture and were lowest when at least 80% pine chips were used as bedding (447.3 and 431.0 mg m, respectively for 80 and 100% pine chip bedding). The highest NH concentrations were observed at Day 28. The highest concentration of TRS was observed when 100% pine chips were used as bedding (11.4 µg m), with high concentrations occurring between Days 7 and 14, and again at Day 35. Greenhouse gases were largely unaffected by bedding material but CH and CO concentrations increased as the bedded packs aged and NO concentrations were highly variable throughout the incubation. We conclude that a mixture of bedding material that contains 30 to 40% pine chips may be the ideal combination to reduce both NH and TRS emissions. All gas concentrations increased as the bedded packs aged, suggesting that frequent cleaning of facilities would improve air quality in the barn, regardless of bedding materials used.

Effect of proximity to a cattle feedlot on Escherichia coli O157:H7 contamination of leafy greens and evaluation of the potential for airborne transmission.

The impact of proximity to a beef cattle feedlot on Escherichia coli O157:H7 contamination of leafy greens was examined. In each of 2 years, leafy greens were planted in nine plots located 60, 120, and 180 m from a cattle feedlot (3 plots at each distance). Leafy greens (270) and feedlot manure samples (100) were collected six different times from June to September in each year. Both E. coli O157:H7 and total E. coli bacteria were recovered from leafy greens at all plot distances. E. coli O157:H7 was recovered from 3.5% of leafy green samples per plot at 60 m, which was higher (P < 0.05) than the 1.8% of positive samples per plot at 180 m, indicating a decrease in contamination as distance from the feedlot was increased. Although E. coli O157:H7 was not recovered from air samples at any distance, total E. coli was recovered from air samples at the feedlot edge and all plot distances, indicating that airborne transport of the pathogen can occur. Results suggest that risk for airborne transport of E. coli O157:H7 from cattle production is increased when cattle pen surfaces are very dry and when this situation is combined with cattle management or cattle behaviors that generate airborne dust. Current leafy green field distance guidelines of 120 m (400 feet) may not be adequate to limit the transmission of E. coli O157:H7 to produce crops planted near concentrated animal feeding operations. Additional research is needed to determine safe set-back distances between cattle feedlots and crop production that will reduce fresh produce contamination.

Use of wood-based materials in beef bedded manure packs: 1. Effect on ammonia, total reduced sulfide, and greenhouse gas concentrations.

The objectives of this study were to determine the effect of using corn stover or three different wood-based bedding materials (kiln-dried pine wood chips, dry cedar chips, or green cedar chips) on airborne concentrations of NH, total reduced sulfides (TRS), CO, CH, and NO above lab-scaled bedded manure packs. Four bedded packs of each bedding material were maintained for two 42-d periods. Airborne NH, TRS, CO, CH, and NO were measured weekly. Bedded packs containing dry or green cedar had lower concentrations of NH (350.8 and 357.3 mg m, respectively; < 0.05) than bedded packs containing pine chips or corn stover (466.0 and 516.7 mg m, respectively). Airborne CO was also lower from bedded packs containing dry and green cedar (1343.7 and 1232.3 mg m, respectively; < 0.001) compared with bedded packs containing pine chips or corn stover (2000.2 and 1659.8 mg m, respectively). Air samples from bedded packs containing green cedar chips had a higher ( < 0.01) concentration of CH than bedded packs containing dry cedar chips, corn stover, or pine chips at Day 35 and 42. Initially, TRS concentration was similar among all bedding materials; at 28 to 42 d, TRS was higher ( < 0.001) from bedded packs containing the cedar products. Airborne NO was similar ( = 0.51) for all bedding materials. Pine chips and cedar products can be adequate substitutes for corn stover in deep-bedded barns, but cedar bedding may need to be removed more frequently.

Use of wood-based materials in beef bedded manure packs: 2. Effect on odorous volatile organic compounds, odor activity value, , and nutrient concentrations.

The objectives of this study were to determine the effects of three types of wood-based bedding materials (kiln-dried pine wood chips, dry cedar chips, and green cedar chips) and corn stover on the concentration of odorous volatile organic compounds (VOCs) and total in bedded pack material. Four bedded packs of each bedding material were maintained for two 42-d periods ( = 32; eight replicates/bedding material). Straight- and branched-chained fatty acids, aromatic compounds, and sulfide compounds were measured from the headspace above each bedded pack. Green cedar bedding had the highest concentration of odorous VOCs, and pine chip bedding had the lowest ( < 0.01). Calculated odor activity values were highest for green cedar bedding, followed by dry cedar, corn stover, and pine chip bedding. As the bedded packs aged, the concentration of odorous VOCs increased, particularly in the bedded packs containing green cedar chips and dry cedar chips. Total concentrations increased from Days 0 to 21 and then began to decline and were similar among all bedding materials ( < 0.10). Results of this study indicate that producers using a long-term bedded pack management in their facility may benefit from using pine chips because they do not appear to increase odor over time. Cedar-based bedding materials may be better suited for a scrape-and-haul system, where the bedded pack is removed after 1 or 2 wk. Total concentrations did not differ between any of the four bedding materials over time.

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.

A framework for developing research protocols for evaluation of microbial hazards and controls during production that pertain to the application of untreated soil amendments of animal origin on land used to grow produce that may be consumed raw.

Application of manure or soil amendments of animal origin (untreated soil amendments; UTSAs) to agricultural land has been a long-standing practice to maintain or improve soil quality through addition of organic matter, nitrogen, and phosphorus. Much smaller quantities of these types of UTSAs are applied to land used for food crops than to land used for animal grain and forage. UTSAs can harbor zoonotic enteric pathogens that may survive for extended periods after application. Additional studies are needed to enhance our understanding of preharvest microbial food safety hazards and control measures pertaining to the application of UTSAs especially for land used to grow produce that may be consumed raw. This document is intended to provide an approach to study design and a framework for defining the scope and type of data required. This document also provides a tool for evaluating the strength of existing data and thus can aid the produce industry and regulatory authorities in identifying additional research needs. Ultimately, this framework provides a means by which researchers can increase consistency among and between studies and facilitates direct comparison of hazards and efficacy of controls applied to different regions, conditions, and practices.