Determination of gastrointestinal tract colonization sites from feedlot cattle transiently shedding or super-shedding Escherichia coli O157:H7 at harvest.

AIMS: The objective of the study was to determine levels of Escherichia coli O157:H7 colonization in the gastrointestinal tract (GIT) of naturally shedding cattle shedding the pathogen at low- or super-shedder levels. METHODS AND RESULTS: Over 2 years, feedlot cattle were sampled multiple times for faecal shedding of E. coli O157:H7. Just prior to harvest (1-2 days), animals that were super-shedders (≥104  CFU per gram of faeces) were specifically identified, and based on the longer term screening data, pen cohorts that were low-shedders (years 1 and 2) or chronic-shedders (year 1) were also identified. At harvest, samples were collected from throughout the GIT, including the rectoanal junction (RAJ) for enumeration and enrichment of E. coli O157:H7. The mouth samples exhibited the greatest prevalence for the pathogen, and the abomasum and rumen exhibited the lowest prevalence (P < 0·05). Super-shedders had significantly greater prevalence for all GIT locations except the mouth and abomasum compared to low-shedders, but the super-shedders were the only animals with positive abomasum samples. Samples from the super-shedders were enumerable for most GIT locations, and the rectum and RAJ locations were the only locations that were significantly greater than other locations (P < 0·05). CONCLUSIONS: Across all animals naturally exposed to E. coli O157:H7, the risk of ingestion is high, but rumen and abomasum are potential barriers to passage. In super-shedders, the passage through the GIT was greater, allowing colonization in the rectum and at the RAJ. SIGNIFICANCE AND IMPACT OF THE STUDY: Escherichia coli O157:H7 low-shedding cattle had lower pathogen levels throughout the GIT, indicating intrinsic GIT factors to these cattle may reduce pathogen passage through the GIT, including the abomasum, and minimize risk of RAJ colonization.

The effects of monensin in diets fed to finishing beef steers and heifers on growth performance and fecal shedding of Escherichia coli O157:H7.

Two experiments were conducted to study the effects of monensin dose on growth performance and O157:H7 shedding in finishing beef cattle. In Exp. 1, 198 heifers (298 ± 1.1 kg BW) were allocated to 1 of 2 treatments consisting of 1) 200 mg/heifer daily of monensin and 2) 400 mg/heifer daily of monensin and fed for 151 d. In Exp. 2, 199 steers (430 ± 1.9 kg BW) were stratified by BW and allocated to 1 of 2 treatments consisting of 1) 0 mg/steer daily of monensin and 2) 400 mg/steer daily of monensin and fed for 128 d. For both experiments, there were 4 pen replicates per treatment. For Exp. 1 and Exp. 2, the model included the fixed effect of treatment for growth performance measures and the fixed effects of treatment, time, and treatment × time interaction, respectively, for O157:H7 shedding. In Exp. 1, final BW was 1.9% greater for heifers fed 400 mg/d monensin than for heifers fed 200 mg/d monensin ( = 0.05). Furthermore, ADG was 4.9% greater ( = 0.05) and G:F was 3.1% greater ( = 0.04) when the heifers were fed 400 mg/d monensin vs. 200 mg/d monensin. Pen prevalence for O157:H7 ( = 0.96) and the percentage of animals in the pen shedding O157:H7 at enumerable levels ( = 0.82) did not differ between heifers fed 200 mg/d monensin and heifers fed 400 mg/d monensin over the 4 sampling periods. For Exp. 2, steers fed the supplement containing monensin had a 1.9% greater final BW ( = 0.04) and a 5.2% greater ADG ( = 0.02) than steers fed a control supplement without monensin. No differences in DMI or G:F were noted across the treatments ( ≥ 0.14). O157:H7 percentage of enumerable cattle within the pen was greater for the steers fed monensin than the control steers not fed monensin than the control steers not fed monensin ( = 0.02) over the 4 sampling periods. However, the percentage of animals in the pen shedding O157:H7 (prevalence positive) did not differ between treatments ( = 0.18), nor did the average fecal counts ( = 0.45). In conclusion, feeding a higher dose (400 mg/d) of monensin improved final BW and ADG compared with a low dose of monensin or a no-monensin control in steers and heifers across multiple years. The percentage of animals shedding O157:H7 at enumerable levels was greater for steers fed the monensin supplement than for steers fed the control supplement, yet the presence of monensin, irrespective of the dose, did not affect the percentage of animals in the pen shedding O157:H7.

The impact of the bovine faecal microbiome on Escherichia coli O157:H7 prevalence and enumeration in naturally infected cattle.

AIMS: The objective of this study was to determine if the faecal microbiome has an association with Escherichia coli O157:H7 prevalence and enumeration. METHODS AND RESULTS: Pyrosequencing analysis of faecal microbiome was performed from feedlot cattle fed one of three diets: (i) 94 heifers fed low concentrate (LC) diet, (ii) 142 steers fed moderate concentrate (MC) diet, and (iii) 132 steers fed high concentrate (HC) diet. A total of 322 585 OTUs were calculated from 2,411,122 high-quality sequences obtained from 368 faecal samples. In the LC diet group, OTUs assigned to the orders Clostridiales and RF39 (placed within the class Mollicutes) were positively correlated with both E. coli O157:H7 prevalence and enumeration. In the MC diet group, OTUs assigned to Prevotella copri were positively correlated with both E. coli O157:H7 prevalence and enumeration, whereas OTUs assigned to Prevotella stercorea were negatively correlated with both E. coli O157:H7 prevalence and enumeration. In both the MC diet group and the HC diet group, OTUs assigned to taxa placed within Clostridiales were both positively and negatively correlated with both E. coli O157:H7 prevalence and enumeration. However, all correlations were weak. In both the MC diet group and the HC diet group, stepwise linear regression through backward elimination analyses indicated that these OTUs were significantly correlated (P < 0·001) with prevalence or enumeration, explaining as much as 50% of variability in E. coli O157:H7 prevalence or enumeration. CONCLUSIONS: Individual colonic bacterial species have little impact on E. coli O157:H7 shedding but collectively groups of bacteria were strongly associated with pathogen shedding. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial groups in the bovine colon may impact faecal shedding of the zoonotic pathogen E. coli O157:H7, and manipulation of the intestinal microbiota to alter these bacteria may reduce shedding of this pathogen and foodborne illnesses.

Relationship of glucocorticoids and hematological measures with feed intake, growth, and efficiency of finishing beef cattle.

The objective of this experiment was to determine the association of glucocorticoids and markers for immune status in finishing beef steers and heifers with DMI, growth, and efficiency. Steers ( = 127) and heifers ( = 109) were individually fed a finishing ration for 84 d with BW measured every 21 d. Blood samples were collected via jugular venipuncture for metabolite (glucose and lactate) and cortisol analysis and rectal grab samples of feces were collected for corticosterone analysis on d 83 of the experiment. Plasma cortisol was not correlated to DMI ( = -0.08, > 0.05) or fractional DMI (g DMI/kg BW; = -0.03, > 0.05) but was negatively correlated with ADG ( = -0.17, < 0.01) and G:F ( = -0.20, < 0.01) and positively correlated to residual feed intake (RFI; = 0.14, < 0.05). Fecal corticosterone was positively correlated to fractional DMI ( = 0.15, < 0.05) and RFI ( = 0.23, < 0.01) and negatively correlated to G:F ( = -0.18, < 0.01). Using a mixed model analysis, none of the metabolites or hormones were associated with DMI ( > 0.05) but fecal corticosterone was positively associated with fractional DMI only in heifers ( = 0.04). Plasma lactate ( < 0.01) was and plasma cortisol ( < 0.10) tended to be negatively associated with ADG. Plasma cortisol ( < 0.05) and fecal corticosterone tended ( < 0.10) to be negatively associated with G:F. Fecal corticosterone was positively associated with RFI in heifers ( < 0.04). In a mixed model analysis, total leukocyte count was positively associated with ADG ( < 0.04) and tended to be positively associated with G:F ( < 0.06). Among leukocyte subtypes, neutrophil count was positively associated with ADG in steers ( < 0.02) and monocytes were positively associated with ADG in heifers ( < 0.03). Lymphocyte counts (LY) in steers were negatively associated with DMI ( = 0.03) and fractional DMI ( < 0.03). In heifers, LY tended to be positively associated with DMI ( < 0.09) and fractional DMI ( < 0.06). Lymphocyte count was also positively associated with ADG ( < 0.01) and G:F ( = 0.05) in heifers. The association of production traits with immune status seems to be different between steers and heifers. There was a stronger relationship of cortisol than fecal corticosterone to feed efficiency measures, suggesting that cortisol concentrations could be a better marker for feed efficiency traits than fecal corticosterone concentrations.

Effect of lysozyme or antibiotics on faecal zoonotic pathogens in nursery pigs.

AIMS: The objective of this study was to determine the effect of lysozyme and antibiotics on zoonotic pathogen shedding in faeces from nursery pigs housed without and with an indirect disease challenge. METHODS AND RESULTS: Two replicates of approximately 650 pigs each were weaned and randomly assigned to one of 24 pens in either a nursery room that had been fully disinfected or a nursery room left unclean. Pigs were randomly assigned to control diet (Control), control diet + antibiotics (Antibiotic; chlortetracycline and tiamulin), or control diet + lysozyme (Lysozyme; 100 mg kg(-1) diet). Rectal swab samples were collected on day 0 and 28 of treatment, and enriched and cultured for Campylobacter spp. and shiga-toxigenic Escherichia coli (STEC). Enrichments from rectal swab samples also were analysed for presence of enterohaemorrhagic E. coli (EHEC) virulence genes (hlyA, eae, stx1 and stx2). Room hygiene had little effect on day 28 results. Percentage of samples culture positive for Campylobacter spp. was lowest for lysozyme diets (P < 0·01), but similar for control and antibiotic diets (43·2, 83·7, and 84·8 respectively). Diet had little effect on the EHEC virulence genes hlyA or eae (P > 0·1), but there was a tendency for fewer samples positive for stx1/stx2 in antibiotic or lysozyme diet groups (P < 0·07) compared to control diet (1·2, 2·1 and 5·8% respectively). Salmonella spp. and specific STEC types tested were rarely detected in the study. CONCLUSIONS: In nursery swine, room hygiene had little effect on pathogen shedding. Dietary chlortetracycline and tiamulin did not reduce pathogen shedding but dietary lysozyme reduced faecal shedding of Campylobacter. SIGNIFICANCE AND IMPACT OF THE STUDY: Lysozyme can effectively replace antibiotics in the diet of nursery swine and can be effective for pathogen control.

Evaluation of essential oils in beef cattle manure slurries and applications of select compounds to beef feedlot surfaces to control zoonotic pathogens.

AIMS: To evaluate natural terpene compounds for antimicrobial activities and determine whether these compounds could be used to control microbial activities and pathogens in production animal facilities. METHODS AND RESULTS: Thymol, geraniol, glydox, linalool, pine oil, plinol and terpineol were tested in laboratory studies for ability to control the production of odorous volatile fatty acid compounds and reduce pathogen levels in manure slurry preparations. Thymol is a terpene phenolic compound and was most effective for reducing fermentation products and pathogen levels (P < 0.05), followed by the extracts linalool, pine oil and terpineol, which are terpene alcohols. Select compounds thymol, linalool and pine oil were further evaluated in two separate studies by applying the agents to feedlot surfaces in cattle pens. Feedlot surface material (FSM; manure and soil) was collected and analysed for fermentation products, levels of coliforms and total Escherichia coli, and the presence of E. coli O157:H7, Campylobacter, Salmonella, Listeria and L. monocytogenes. The reduction in fermentation products but not pathogens was dependent on the moisture present in the FSM. Treatment with 2000 ppm thymol reduced the prevalence of E. coli O157:H7 but not Listeria. In a separate study, treatment with 4000 ppm pine oil reduced E. coli O157:H7, Listeria and Campylobacter (P < 0.05). Linalool was tested at two levels (2000 and 4000 ppm) and did not affect pathogen levels at either concentration. CONCLUSIONS: Natural compounds bearing terpenes can control pathogenic bacteria in treated manures and when applied to the feedlot surface in production cattle systems. Pine oil is a cheaper alternative to thymol and may be a useful treatment for controlling pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: The control of bacterial pathogens in animal productions systems is an important step in preharvest food safety. Waste products, such as pine oil extract, from the pulp wood industry may have application for treating feedlot pens and manures to reduce the pathogen load.

Investigation of bacterial diversity in the feces of cattle fed different diets.

The objective of this study is to investigate individual animal variation of bovine fecal microbiota including as affected by diets. Fecal samples were collected from 426 cattle fed 1 of 3 diets typically fed to feedlot cattle: 1) 143 steers fed finishing diet (83% dry-rolled corn, 13% corn silage, and 4% supplement), 2) 147 steers fed late growing diet (66% dry-rolled corn, 26% corn silage, and 8% supplement), and 3) 136 heifers fed early growing diet (70% corn silage and 30% alfalfa haylage). Bacterial 16S rRNA gene amplicons were determined from individual fecal samples using next-generation pyrosequencing technology. A total of 2,149,008 16S rRNA gene sequences from 333 cattle with at least 2,000 sequences were analyzed. Firmicutes and Bacteroidetes were dominant phyla in all fecal samples. At the genus level, Oscillibacter, Turicibacter, Roseburia, Fecalibacterium, Coprococcus, Clostridium, Prevotella, and Succinivibrio were represented by more than 1% of total sequences. However, numerous sequences could not be assigned to a known genus. Dominant unclassified groups were unclassified Ruminococcaceae and unclassified Lachnospiraceae that could be classified to a family but not to a genus. These dominant genera and unclassified groups differed (P < 0.001) with diets. A total of 176,692 operational taxonomic units (OTU) were identified in combination across all the 333 cattle. Only 2,359 OTU were shared across 3 diet groups. UniFrac analysis showed that bacterial communities in cattle feces were greatly affected by dietary differences. This study indicates that the community structure of fecal microbiota in cattle is greatly affected by diet, particularly between forage- and concentrate-based diets.

Effects of antimicrobials fed as dietary growth promoters on faecal shedding of Campylobacter, Salmonella and shiga-toxin producing Escherichia coli in swine.

AIMS: To determine whether antimicrobials commonly used in swine diets affect zoonotic pathogen shedding in faeces. METHODS AND RESULTS: Barrows (n = 160) were sorted into two treatments at 10 weeks of age (week 0 of the study), and fed growing, grow finishing and finishing diets in 4-week feeding periods. For each feeding phase, diets were prepared without (A-) and with (A+) dietary antimicrobials (chlortetracycline, 0-8 week; bacitracin, 9-12 week) typical of the United States. At week 0, 4, 8, 9, 10 and 12 of the study, faecal swabs or grabs were collected for analyses. Campylobacter spp. was absent at week 0, but prevalence increased over time with most isolates being identified as Campylobacter coli. When chlortetracycline was used in A+ diets (week 4 and 8), prevalence for Campylobacter spp., pathogenic Escherichia coli O26 and stx genes was lower in faeces. On week 12 after the shift to bacitracin, Campylobacter spp. and stx genes were higher in faeces from piglets fed A+ diet. Pathogenic E. coli serogroups O103 and O145 were isolated throughout the study and their prevalence did not differ due to diet. Pathogenic E. coli serogroups O111 and O121 were never found in the piglets, and Salmonella spp. prevalence was low. CONCLUSIONS: In production swine, growing diets with chlortetracycline may have reduced pathogen shedding compared with the A-growing diets, whereas finishing diets with bacitracin may have increased pathogen shedding compared with the A-finishing diet. SIGNIFICANCE AND IMPACT OF THE STUDY: Inclusion of antimicrobials in the diet can affect zoonotic pathogen shedding in faeces of swine.

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

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

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

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

Odorant production and persistence of Escherichia coli in manure slurries from cattle fed zero, twenty, forty, or sixty percent wet distillers grains with solubles.

Corn ethanol production removes starch and concentrates the remaining nutrients, including CP and minerals. When wet distillers grains with solubles (WDGS) are fed to cattle in place of corn, CP and minerals often exceed dietary needs. This may increase N emission, P run-off, and odor production. These variables are evaluated in this study. Crossbred steers (n = 160; 434 +/- 8 kg) were assigned in a completely randomized block design to 9 x 9 m pens with concrete floor (10 animals/pen; 4 pens/treatment). Steers were fed a finishing diet that contained 0, 20, 40, or 60% WDGS on a DM basis, and provided 13.3, 15.5, 20.6, or 24.9% CP, respectively. Two kilograms of manure slurry (14 to 23% DM) were collected from each pen monthly (Aug. 20, Sep. 24, and Oct. 22). Samples were analyzed immediately for odorants, DM, pH, NH(3), total alcohol, l-lactate, and concentrations of generic Escherichia coli. After incubation of the samples at 22 degrees C for 2, 4, 7, 10, 15, 21, and 28 d, samples were analyzed for methane production in addition to the above characteristics. Before incubation, NH(3), H(2)S, indole, phenol, isovalerate, isobutyrate, and acetate increased (P < 0.01) with increasing amounts of WDGS in the diet. Other odorants, including skatole, caproate, valerate, butyrate, and propionate, were greater (P < 0.01) in manure slurries from cattle fed 20 or 40% WDGS, compared to 0% WDGS. The l-lactate was greater (P < 0.01) in slurries from cattle fed 0% WDGS (447 mu mol/g of DM) compared with the other treatment slurries (14 to 15 mu mol/g of DM). After incubation, l-lactate contributed to lowered slurry pH (6.3, 7.1, 7.6, and 8.2, respectively, for 0, 20, 40, and 60% WDGS), which inhibited microbial fermentation, E. coli persistence, and methane production. Because of the favorable, more neutral pH in the 40 and 60% WDGS slurries, many of the odorant compounds were rapidly converted to methane during a 28-d static incubation. Escherichia coli O157:H7 inoculated into subsamples of the manure slurries exhibited behavior similar to that of naturally present generic E. coli, surviving in greater numbers longer (P < 0.05) in 20 and 40% WDGS slurries than in 0% WDGS. These data indicate feeding WDGS can increase odorants in manure slurries and extend the persistence of E. coli.

The influence of oscillating dietary protein concentrations on finishing cattle. I. Feedlot performance and odorous compound production.

We hypothesized that oscillation of the dietary CP concentration, which may improve N retention of finishing beef steers, would reduce production of manure odor compounds and total N inputs while yielding comparable performance. Charolais-sired steers (n = 144; 303 +/- 5 kg of initial BW) were used in a completely randomized block design (6 pens/treatment). The steers were fed to 567 kg of BW on the following finishing diets, which were based on dry-rolled corn: 1) low (9.1% CP), 2) medium (11.8% CP), 3) high (14.9% CP), or 4) low and high oscillated on a 48-h interval for each feed (oscillating). Steers fed low tended (P = 0.08) to have less DMI (7.80 kg/d) than steers fed medium (8.60 kg/d) or oscillating (8.67 kg/d), but not less than steers fed high (8.12 kg/d). Daily N intake was greatest (P < 0.01) for steers fed high (189 g), intermediate for medium (160 g) and oscillating (164 g), and least for low (113 g). The ADG was lower (P < 0.01) for steers fed low (1.03 kg) than for those fed medium (1.45 kg), high (1.45 kg), or oscillating (1.43 kg). Similarly, steers fed low had a lower adjusted fat thickness (P < 0.01) and yield grade (P = 0.05) and tended (P = 0.10) to have less marbling than steers fed the other 3 diets. In slurries with feces, urine, soil, and water, incubated for 35 d, nonsoluble CP was similar among slurries from steers fed medium, high, or oscillating, but was less (P < 0.01) in slurries from steers fed low. However, throughout the incubation period, slurries from steers fed high or oscillating had greater (P < 0.01) concentrations of total aromatics and ammonia than those from steers fed low or medium. Also, the slurries from steers fed oscillating had greater (P < 0.01) concentrations of branched-chain VFA than manure slurries from steers fed any of the other diets. These data indicate that although there is no apparent alteration in the performance of finishing steers fed diets with oscillation of the dietary protein, there may be undesirable increases in the production of compounds associated with malodor.

Influence of genotype and diet on steer performance, manure odor, and carriage of pathogenic and other fecal bacteria. I. Animal performance.

Although Brahman crosses constitute a large portion of US beef cattle, little information is available on their response to diverse feed resources compared with Bos taurus steers. Thus, the objectives were to evaluate genotype and diet effects on steer performance during the growing period and subsequent response to a high grain diet during the finishing period. Fifty-one steers [0 (15), 1/4 (20), 1/2 (7), and 3/4 Brahman (9), with the remaining proportion being MARC III] were allotted to 8 pens. Beginning on December 2, steers were individually fed chopped bromegrass hay (n = 26; DM = 85%, CP = 9.5%, ME = 2.19 Mcal/kg) or a corn silage-based diet (n = 25; DM = 51%, CP = 11.9%, ME = 2.75 Mcal/kg) for 119 d. All steers were then fed a high corn diet (DM = 79%, CP = 11.7%, ME = 3.08 Mcal/kg) to a target BW of 560 kg (176 d). Data were analyzed by ANOVA, with genotype, growing diet, and the 2-way interaction included. The interaction was not significant (P > 0.25). The MARC III and 1/2 Brahman steers weighed more (P < 0.01) than 1/4 or 3/4 Brahman steers initially and at the end of the growing period. Weight of bromegrass-fed (325 kg) steers was less than that of corn silage-fed (384 kg) steers at the end of the growing period. Steer ADG and intake of DM, CP, and ME were less (P = 0.087 to 0.001) for 1/4 and 3/4 Brahman than for 0 or 1/2 Brahman steers during growing, finishing, and total, but efficiency of gain did not differ (P > 0.10). Carcass weight, marbling score, quality grade (P < 0.05), and kidney fat (P = 0.06) differed among genotypes. Daily DMI (6.91 vs. 7.06 kg) was similar, but CP (0.66 vs. 0.84 kg) and ME (15.2 vs. 19.4 Mcal) intake of bromegrass fed was less (P = 0.001) than those of corn silage-fed steers. Values for DMI/gain (22.3 vs. 7.43 kg/kg), CP intake/gain (2.13 vs. 0.88 kg/kg), and ME intake/gain (48.8 vs. 20.4 Mcal/kg) were greater (P < 0.001) in bromegrass-fed than corn silage-fed steers. Over the total study, ADG was lower (0.96 vs. 1.01 kg), and DMI (7.82 vs. 7.19 kg), DMI/gain (8.21 vs. 7.10 kg/kg), and ME intake/gain (22.6 vs. 20.9 Mcal/kg) were greater (P < 0.05) in bromegrass-fed than in corn silage-fed steers. Carcass weight, dressing percent, adjusted backfat, and yield grade (P < 0.05) were greater for corn silage-fed than for bromegrass-fed steers. Feed intake and performance, but not efficiency, differed among these genotypes. Compensatory performance during finishing was insufficient to overcome reduced performance during the growing period.

Influence of genotype and diet on steer performance, manure odor, and carriage of pathogenic and other fecal bacteria. II. Pathogenic and other fecal bacteria.

This study assessed the influence of cattle genotype and diet on the carriage and shedding of zoonotic bacterial pathogens and levels of generic Escherichia coli in feces and ruminal contents of beef cattle during the growing and finishing periods. Fifty-one steers of varying proportions of Brahman and MARC III [0 (15), 1/4 (20), 1/2 (7), and 3/4 Brahman (9)] genotypes were divided among 8 pens, such that each breed type was represented in each pen. Four pens each were assigned to 1 of 2 diets [100% chopped bromegrass hay or a diet composed primarily of corn silage (87%)] that were individually fed for a 119-d growing period, at which time the steers were switched to the same high-concentrate, corn-based finishing diet and fed to a target weight of 560 kg. Feces or ruminal fluid were collected and analyzed at alternating intervals of 14 d or less. Generic E. coli concentrations in feces or ruminal fluid did not differ (P > 0.10) by genotype or by growing diet in the growing or finishing periods. However, the concentrations in both feces and ruminal fluid increased in all cattle when switched to the same high-corn diet in the finishing period. There was no effect (P > 0.25) of diet or genotype during either period on E. coli O157 shedding in feces. Forty-one percent of the steers were positive for Campylobacter spp. at least once during the study, and repeated isolations of Campylobacter spp. from the same steer were common. These repeated isolations from the same animals may be responsible for the apparent diet (P = 0.05) and genotype effects (P = 0.02) on Campylobacter in feces in the finishing period. Cells bearing stx genes were detected frequently in both feces (22.5%) and ruminal fluid (19.6%). The number of stx-positive fecal samples was greater (P < 0.05) for 1/2 Brahman steers (42.9%) than for 1/4 Brahman (25.0%) or 3/4 Brahman steers (22.2%), but were not different compared with MARC III steers (38.3%). The greater feed consumption of 1/2 Brahman and MARC III steers may have resulted in greater starch passage into the colon, accompanied by an increase in fecal bacterial populations, which may have further improved the ability to detect stx genes in these cattle. There was no correlation between either ADG or daily DMI and the number of positive samples of E. coli O157, Campylobacter spp., or stx genes, which agrees with our current understanding that these microorganisms occur commonly in, and with no measurable detriment to, healthy cattle.

Influence of genotype and diet on steer performance, manure odor, and carriage of pathogenic and other fecal bacteria. III. Odorous compound production.

Three beef cattle diets were assessed for their potential to produce odorous compounds from cattle feces excreted during the growing and finishing periods. Eight pens containing 51 steers of varying proportions of Brahman and MARC III genotypes were fed either a chopped bromegrass hay diet or a corn silage diet for a 119-d growing period. After the growing period, all steers were switched to the same high-corn finishing diet (high corn) and fed to a target weight of 560 kg (finishing period). Fecal slurries were prepared from a composite of fresh fecal pats collected in each pen during both periods and incubated anaerobically. In additional flasks, starch, protein, or cellulose was added to the composite fecal subsamples to determine the preferred substrates for fermentation and odorous compound production. The content and composition of the fermentation products varied both initially and during the incubation, depending on the diet fed to the steers. The corn silage and high corn feces had the greater initial content of VFA (381.0 and 524.4 micromol/g of DM, respectively) compared with the bromegrass feces (139.3 micromol/g of DM) and accumulated more VFA than the bromegrass feces during the incubation. l-Lactic acid and VFA accumulation in the high corn and corn silage feces was at the expense of starch, based on starch loss and the production of straight-chain VFA. In the bromegrass feces, accumulation of branched-chain VFA and aromatic compounds and the low starch availability indicated that the protein in the feces was the primary source for odorous compound production. Substrate additions confirmed these conclusions. We conclude that starch availability was the primary factor determining accumulation and composition of malodorous fermentation products, and when starch was unavailable, fecal microorganisms utilized protein.

Incorporation of thymol into corncob granules for reduction of odor and pathogens in feedlot cattle waste.

Confined animal feeding operations can be a source of odor emissions, global warming gases, water pollution, and food contamination. Laboratory studies have indicated that plant oils with antimicrobial activity can be used to control pathogens and odor emissions from cattle and swine wastes. However, these oils are aromatic and may volatilize when applied topically. Our objectives were to evaluate the volatility of thymol from a feedlot surface and the effectiveness of topically applying thyme oil (2.5% thymol), incorporated into corncob granules and added once per week, to control odor emissions and total coliforms in feedlot manure. In the first study, thymol either volatilized or was degraded within 28 d after topical application. In a second study, thyme oil (2.5% thymol) was incorporated into corncobs and applied to pen surfaces weekly. Manure samples from 6 locations in each pen were collected from 3 untreated and 3 thymol-corncob-treated pens (15 x 150 m; fifty 400-kg cattle/pen), 3 times per week for 8 wk. Samples were analyzed for thymol concentration, total VFA, branched-chain VFA, aromatic compounds, and the number of Escherichia coli and total coliform bacteria. Over the 8 wk, with the exception of wk 7, the desired thymol concentration of 15 to 20 micromol/g DM was maintained in the manure. Concentrations of VFA and branched chain-VFA increased over time in untreated and treated pens. However, the rate of VFA accumulation in treated pens (7.5 +/- 1.3 micromol.g DM(-1).wk(-1)) was less (P < 0.01) than the rate of accumulation in untreated pens (18.0 +/- 2.1 micromol.g DM(-1).wk(-1)). Likewise, the rate of branched-chain VFA accumulation in treated pens (0.31 +/- 0.04 micromol.g DM(-1).wk(-1)) was less (P < 0.01) than in untreated pens (0.55 +/- 0.06 micromol.g DM(-1).wk(1)). The concentrations of E. coli in treated pens (2.9 +/- 1.2 x 10(5) cfu.g DM(-1)) were 91% less (P < 0.04) than in untreated pens (31.1 +/- 4.0 x 10(5) cfu.g DM(-1)). Similarly, concentrations of coliforms in treated pens (3.7 +/- 1.3 x 10(5) cfu.g DM(-1)) were 89% less (P < 0.04) than those of untreated pens (35.3 +/- 4.2 x 10(5) cfu.g DM(-1)). These results indicate that odor emissions and total coliforms can be reduced in feedlot manure with a once per week application of thymol incorporated in a granular form. However, corncobs are bulky, and other granular carriers with a greater carrying capacity for thyme oil should be explored.

Effects of common forage phenolic acids on Escherichia coli O157:H7 viability in bovine feces.

Ruminant animals are carriers of Escherichia coli O157:H7, and the transmission of E. coli O157:H7 from cattle to the environment and to humans is a concern. It is unclear if diet can influence the survivability of E. coli O157:H7 in the gastrointestinal system or in feces in the environment. Feces from cattle fed bromegrass hay or corn silage diets were inoculated with E. coli O157:H7, and the survival of this pathogen was analyzed. When animals consumed bromegrass hay for <1 month, viable E. coli O157:H7 was not recovered after 28 days postinoculation, but when animals consumed the diet for >1 month, E. coli O157:H7 cells were recovered for >120 days. Viable E. coli O157:H7 cells in feces from animals fed corn silage were detected until day 45 and differed little with the time on the diet. To determine if forage phenolic acids affected the viability of E. coli O157:H7, feces from animals fed corn silage or cracked corn were amended with common forage phenolic acids. When 0.5% trans-cinnamic acid or 0.5% para-coumaric acid was added to feces from silage-fed animals, the E. coli O157:H7 death rate was increased significantly (17-fold and 23-fold, respectively) compared to that with no addition. In feces from animals fed cracked corn, E. coli O157:H7 death rates were increased significantly with the addition of 0.1% and 0.5% trans-cinnamic acid (7- and 13-fold), 0.1% and 0.5% p-coumaric acid (3- and 8-fold), and 0.5% ferulic acid (3-fold). These data suggest that phenolic acids common to forage plants can decrease viable counts of E. coli O157:H7 shed in feces.

Stationary-phase acid resistance and injury of recent bovine Escherichia coli O157 and non-O157 biotype I Escherichia coli isolatest.

Stationary-phase acid resistance and the induction of acid resistance were assessed for recent bovine carcass isolates of Escherichia coli, including 39 serotype O157 strains and 20 non-O157 strains. When grown to stationary phase in the absence of glucose and without prior acid exposure, there was a range of responses to a pH challenge of 6 h at pH 2.5. However, populations of 53 of the 59 E. coli isolates examined were reduced by less than 2.00 log CFU/ml, and populations of 24 of these isolates were reduced by less than 1.00 log CFU/ml. In contrast, there was little variation in population reductions when the E. coli were grown with glucose and preadapted to acidic conditions. With few exceptions, acid adaptation improved survival to the acid challenge, with 57 of the 59 isolates exhibiting a log reduction of less than 0.50. Differences in acid resistance or the ability to adapt to acidic conditions between E. coli O157:H7 and non-O157 commensal E. coli were not observed. However, we did find that the E. coli O157 were disposed to greater acid injury after the low pH challenge than the non-O157 E. coli, both for cells that were and were not adapted to acidic conditions before the challenge. The enhancement of low pH survival after acid adaptation that was seen among these recent natural isolates of E. coli O157 further supports the idea that the previous environment of this pathogen should be a consideration when designing microbial safety strategies for foods preserved by low pH and acid.

Effect of different levels of beef bacterial microflora on the growth and survival of Escherichia coli O157:H7 on beef carcass tissue.

The influence of various levels of endogenous beef bacterial microflora on the growth and survival of Escherichia coli O157:H7 on bovine carcass surface tissue was investigated. Bacterial beef microflora inoculum was prepared by enriching and harvesting bacteria from prerigor lean bovine carcass tissue (BCT) and was inoculated onto UV-irradiated prerigor BCT at initial levels of 10(5), 10(4), 10(3), and <10(3) CFU/cm2. Additional control BCT was inoculated with sterile H2O. E. coli O157:H7 was inoculated onto all tissues at an initial level of 10(2) CFU/cm2. Following a 48-h incubation at 4 degrees C, BCT was incubated up to 14 days at 4 or 12 degrees C, either aerobically or vacuum packaged. Regardless of the microflora level, there was no substantial growth of E. coli O157:H7 on BCT during storage at 4 degrees C under either aerobic or vacuum-packaged conditions. Instead, viable cell numbers at 4 degrees C remained constant, with no reduction in numbers associated with the different beef microflora levels. E. coli O157:H7 grew on all BCT stored at 12 degrees C, regardless of microflora inoculation treatment, reaching higher populations on aerobic samples than on vacuum-packaged samples in 10 days. However, the presence of the beef microflora did appear to delay the onset of growth or slow the growth of the pathogen, and E. coli O157:H7 counts on BCT without added microflora were generally higher following 7 to 10 days of 12 degrees C storage than those counts on BCT inoculated with beef microflora. These data demonstrate the importance of temperature control during meat handling and storage to prevent the outgrowth of this pathogen and indicate that proper sanitation and processing practices that prevent and reduce contamination of carcasses with E. coli O157:H7 are essential, regardless of background microflora levels.

Evaluation of combination treatment processes for the microbial decontamination of pork trim.

Combination treatment processes for the microbial decontamination of pork trim were developed and evaluated. Lean pork trim tissue (LPT) and fat-covered pork trim tissue (FPT) inoculated with swine feces were treated with intervention processes as follows: (i) control (untreated), (ii) water (15 degrees C, 120 s), (iii) water followed by lactic acid wash (15 degrees C, 75 s), (iv) combination 1 (water plus hot water [65.5 degrees C, 15 s] plus hot air [510 degrees C, 60 s] plus lactic acid), (v) combination 2 (water plus hot water [82.2 degrees C, 15 s] plus hot air [510 degrees C, 75 s] plus lactic acid), and (vi) combination 3 (water plus hot water [82.2 degrees C, 45 s] plus hot air [510 degrees C, 90 s] plus lactic acid). Populations of aerobic bacteria, psychrotrophic bacteria, coliforms, Escherichia coli, and lactic acid bacteria were determined before and after treatment and at days 2 and 7 of 4 degrees C storage. Regardless of the intervention treatment, lower microbial populations were observed on FPT than on LPT immediately after treatment and during the 7-day storage period. Both LPT and FPT treated with water plus lactic acid, combination 1, combination 2, and combination 3 had lower remaining populations of all microbial groups immediately after treatment than did water-treated samples. Populations of aerobic bacteria, coliforms, E. coli, and lactic acid bacteria on either LPT or FPT did not statistically increase during the 7-day storage period. On LPT, populations of psychrotrophic bacteria grew during 4 degrees C storage but remained lower at day 7 on LPT treated by combinations 2 and 3 (2.29 and 1.89 log10 CFU/cm2, respectively) than on LPT treated with water (4.07 log10 CFU/cm2) or water plus lactic acid (3.52 log10 CFU/cm2). Populations of psychrotrophic bacteria remained below detectable levels throughout the 7-day storage on FPT treated with water plus lactic acid or any of the three combination treatments. Treatment of pork trim with any of the combination treatments significantly (P < 0.05) affected the color and emulsion stability of the ground pork. Water and water plus lactic acid were the most favorable treatments in reducing microbial populations on pork trim without affecting the quality attributes of the ground pork.