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.

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.

Development of a UHPLC-MS/MS method for the measurement of chlortetracycline degradation in swine manure.

An ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed capable of simultaneously measuring chlortetracycline (CTC), epi-chlortetracycline, and isochlortetracycline (ICTC), as well as other structurally related tetracyclines in swine manure. A simple sample preparation was used consisting of extraction, dilution, centrifugation, and ultrafiltration. The concentrations of analyte were calculated using d(6)-tetracycline as an internal standard in the matrix-matched standard curve. A solvent gradient resolved the compounds in 3.5 min with an additional 1.5 min of re-equilibration allowing the analyses of a large number of samples in a short period of time. MS/MS was used as the detection method giving analyte confirmation in addition to a large dynamic range and low detection limit. The UHPLC-MS/MS method successfully resolved multiple degradation products of CTC from the complex manure matrix. The method detection limits ranged from 1.9 pg/µL for CTC to 7.3 pg/µL for ICTC, and the calibration curve was linear from 1 to 10,000 pg/µL. The method was tested by measuring CTC and its degradation products as a function of time in incurred swine manure that had been incubated at three different temperatures (22 °C, 38 °C, and 55 °C). CTC concentration at 22 °C decreased 44% after 25 days; greater percentage decreases were observed when the manure was stored at elevated temperatures (96% and 98% for 38 °C and 55 °C, respectively). The concentration of the microbiologically inactivate isomer, ICTC, increased over the incubation period. At 22 °C, ICTC continued to increase through 25 days of incubation; at 38 °C, ICTC concentration plateaued on day 14 while at 55 °C ICTC concentration plateaued on day 7, with concentration increases of 198%, 374%, and 282% for 22 °C, 38 °C, and 55 °C, respectively.

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.

Influence of thymol and a urease inhibitor on coliform bacteria, odor, urea, and methane from a swine production manure pit.

Pathogens, ammonia, odor, and greenhouse gas emissions are serious environmental concerns associated with swine production. This study was conducted in two manure pits (33,000 L each) to determine the influence of 1.5 or 3.0 g thymol L(-1) and 80 mg L(-1) urease inhibitor amendments on urea accumulation, coliform bacteria, odor, and methane emission. Each experiment lasted 18 or 19 d, during which time 30 to 36 250-mL samples (six per day) were withdrawn from underneath each pit and analyzed for urea, thymol, volatile fatty acids, coliform bacteria, and Campylobacter. At the end of each experiment, six 50-g samples from each pit were placed in serum bottles, and gas volume and composition were determined periodically for 28 d. Compared with the control pit, volatile fatty acids production was reduced 64 and 100% for the thymol amendments of 1.5 and 3.0 g L(-1), respectively. Viable coliform cells were reduced 4.68 and 5.88 log10 colony-forming units kg(-1) of slurry for the 1.5 and 3.0 g thymol L(-1), respectively, and Escherichia coli were reduced 4.67 and 5.01 log10 colony-forming units kg(-1) of slurry, respectively. Campylobacter was not detected in the pits treated with thymol, in contrast to 63% of the samples being positive for the untreated pit. Urea accumulated in the treated pits from Day 3 to 6. Total gas production from serum bottles was reduced 65 and 76% for thymol amendments of 1.5 and 3.0 g L(-1), respectively, and methane was reduced 78 and 93%, respectively. These results suggest that thymol markedly reduces pathogens, odor, and greenhouse gas emissions from a swine production facility. The urease inhibitor produced a temporary response in conserving urea.

Carvacrol and thymol reduce swine waste odor and pathogens: stability of oils.

An incomplete anoxic fermentation of livestock waste results in offensive odor emissions. Antimicrobial additives may be useful in controlling odor emissions and pathogens. Natural antimicrobial compounds, carvacrol or thymol at 16.75 mM (2.5 g/l) completely inhibited the production of the offensive odor compounds, isobutyrate, valerate, isovalerate, and cresol, and significantly reduced other short-chain volatile fatty acids and gas emissions from swine waste. Fecal coliforms were reduced from 6.3 x 10(6) to 1.0 x 10(3) cells per ml 2 days after treatment with carvacrol (13.3 mM) and were not detectable within 14 days. Total culturable anaerobic bacteria were reduced from 12.4 x 10(10) to 7.2 x 10(8) cells per ml after 2 days and were suppressed below this level for 28 days. Lactate production was not prevalent in untreated swine waste indicating that the microbial populations differ from those in cattle waste. Carvacrol and thymol were stable in swine waste under anoxic conditions for 62 days with 90 to 95% of the additive being recovered in the waste solids. In conclusion, carvacrol and thymol are not metabolized in anoxic swine waste and they are potentially useful in controlling odor emissions and pathogens in swine waste.