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.

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.

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.

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.

Effect of bedding materials on concentration of odorous compounds and in beef cattle bedded manure packs.

The objectives of this study were to determine the effect of bedding material (corn stover, soybean stover, wheat straw, switchgrass, wood chips, wood shavings, corn cobs, and shredded paper) on concentration of odorous volatile organic compounds (VOC) in bedded pack material and to determine the effect of bedding material on the levels of total in laboratory-scaled bedded manure packs. Four bedded packs of each bedding material were maintained for two 6-wk periods ( = 64). Straight- and branched-chained fatty acids and aromatic compounds were measured. Corn cob bedding had the highest concentration of odorous VOC, and wood shavings had the lowest ( < 0.01). Calculated odor activity values were highest for corn cob bedding and shredded paper and lowest for wood shavings ( < 0.01). concentrations decreased from week to week for all treatments from Week 2 to Week 6. At Week 6, levels in bedded packs with shredded paper were higher ( < 0.05) than bedded packs containing wood shavings, wood chips, or switchgrass ( < 0.05). At Weeks 4, 5, and 6, concentrations in bedded packs with wood shavings were lower ( < 0.05) than bedded packs of all treatments except wood chips. Results of this study indicate that ground corn cobs or shredded paper may increase odor production and shredded paper may increase when used in deep-bedded livestock facilities, whereas wood shavings may have the least impact on air quality and .

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.

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.

Manure odor potential and Escherichia coli concentrations in manure slurries of feedlot steers fed 40% corn wet distillers grains.

This study evaluated feeding 0 and 40% wet distillers grains with solubles (WDGS) diets to cattle and the effects on feedlot manure collected from soil-based pens and incubated for 28 d. Steers (n = 603; 261 +/- 32 kg) were fed in eight pens (15 x 150 m) of 75 to 77 steers per pen. Two consecutive experiments were conducted with WDGS--one in which the corn source fed with WDGS was high-moisture and one in which WDGS was fed with dry-rolled corn. We compared odorants (volatile fatty acids [VFAs], aromatic compounds, NH3, H2S) and persistence of Escherichia coli in feedlot manure slurries stored from 0 to 28 d. From both experiments, manure collected from cattle fed 40% WDGS had lower (P < 0.05) total VFAs, including acetate, propionate, and butyrate, all of which continued to be lower to 28 d. However, these slurries had greater concentrations (P < 0.05) of branched-chained VFAs (isobutyrate and isovalerate), especially after 14 d of incubations. Similarly, p-cresol and skatole concentrations tended to be greater in slurries originating from 40% WDGS diets and increased with incubation time. Indole was initially greater in the slurries from 40% WDGS diets; however, it was metabolized by microbes during incubation. Manure slurries from the 40% WDGS diets had greater quantities of H2S, NH3, and P (P < 0.05). Levels of E. coli in 0 and 40% WDGS manure slurries were similar when high-moisture corn was used in the diets. However, when dry-rolled corn was used, E. coli persisted longer in 40% WDGS manure slurries in comparison to 0% WDGS. Results here support earlier studies that suggest feeding WDGS increases odor emissions, N loss, E. coli survival, and surface water contamination due to greater potential P runoff.

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.

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.

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.

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.

A direct plating method for estimating populations of Escherichia coli O157 in bovine manure and manure-based materials.

Escherichia coli O157:H7 outbreaks associated with produce consumption have brought attention to livestock manures and manure-based soil amendments as potential sources of pathogens for the contamination of these crops. Procedures for enumeration of E. coli O157:H7 are needed to assess the risks of transmission from these manures and their by-products. A direct plating method employing spiral plating onto CHROMagar O157 was investigated for enumeration of E. coli O157:H7 in feedlot surface material, aged bovine manure, bovine manure compost, and manure-amended soil. In studies utilizing samples spiked with a five-strain cocktail of E. coli O157:H7 at levels ranging from 102 to 10(5) CFU/g of sample, there were strong correlations between the observed and predicted levels of this pathogen. Although the addition of 2.5 mg/liter potassium tellurite and 5 mg/liter novobiocin made the medium more restrictive, these amendments enhanced the ability to identify and enumerate E. coli O157:H7 in feedlot surface material, which contained a higher proportion of fresh feces than did the other three sample types and therefore higher levels of interfering bacterial microflora. The spiral plating method was further assessed to determine its ability to enumerate E. coli O157:H7 in naturally contaminated feedlot surface material. Comparison of E. coli O157:H7 counts in feedlot surface material obtained by the spiral plating method and a most probable number technique were well correlated. We conclude that direct spiral plating onto CHROMagar O157 is effective for estimating E. coli O157:H7 levels in a variety of manures and manure-containing sample types to a lower detection limit of 200 CFU/g. The method has application for determining E. coli O157:H7 concentrations in manures and composts before their sale and use as soil amendments and for measuring the effectiveness of manure treatment processes to reduce or inactivate this pathogen.