ABSTRACT The disinfectant and antimicrobial susceptibility profiles of 138 non-O157 Shiga toxin–producing Escherichia coli strains (STECs) from food animals and humans were determined. Antimicrobial resistance (AMR) was moderate (39.1% of strains) in response to 15 antimicrobial agents. Animal strains had a lower AMR prevalence (35.6%) than did human strains (43.9%) but a higher prevalence of the resistance profile GEN-KAN-TET. A decreasing prevalence of AMR was found among animal strains from serogroups O45 > O145 > O121 > O111 > O26 > O103 and among human strains from serogroups O145 > O103 > O26 > O111 > O121 > O45. One animal strain from serogroups O121 and O145 and one human strain from serogroup O26 had extensive drug resistance. A high prevalence of AMR in animal O45 and O121 strains and no resistance or a low prevalence of resistance in human strains from these serogroups suggests a source other than food animals for human exposure to these strains. Among the 24 disinfectants evaluated, all strains were susceptible to triclosan. Animal strains had a higher prevalence of resistance to chlorhexidine than did human strains. Both animal and human strains had a similar low prevalence of low-level benzalkonium chloride resistance, and animal and human strains had similar susceptibility profiles for most other disinfectants. Benzyldimethylammonium chlorides and C10AC were the primary active components in disinfectants DC&R and P-128, respectively, against non-O157 STECs. A disinfectant FS512 MIC ≥ 8 μg/ml was more prevalent among animal O121 strains (61.5%) than among human O121 strains (25%), which may also suggest a source of human exposure to STEC O121 other than food animals. Bacterial inhibition was not dependent solely on pH but was correlated with the presence of dissociated organic acid species and some undissociated acids.

Food producing animals can be reservoirs of Campylobacter , a leading bacterial cause of human foodborne illness. Campylobacter spp. utilize amino acids as major carbon and energy substrates, a process that can be inhibited by thymol and diphenyliodonium chloride (DIC). To determine the effect of these potential additives on feed intake, live weight gain, and gut Campylobacter levels, growing pigs were fed standard grower diets supplemented with or without 0.0067 or 0.0201% thymol or 0.00014 or 0.00042% DIC in a replicated study design. Diets were offered twice daily for 7 days, during which time daily feed intake (mean ± SEM, 2.39 ± 0.06 kg day −1 ) and daily gain (0.62 ± 0.04 kg day −1 ) were unaffected ( P > 0.05) by treatment. Pigs treated with DIC but not thymol tended to have lower rectal Campylobacter levels ( P = 0.07) (5.2 versus 4.2 and 4.4 log CFU g −1 rectal contents for controls and 0.00014% DIC and 0.00042% DIC, respectively; SEM = 0.26). However, DIC or thymol treatments did not affect ( P > 0.05) ileal or cecal Campylobacter (1.6 ± 0.17 and 4.5 ± 0.26 log CFU g −1 , respectively), cecal total culturable anaerobes (9.8 ± 0.10 log CFU g −1 ), or accumulations of major fermentation end products within collected gut contents. These results suggest that thymol and DIC were appreciably absorbed, degraded, or otherwise made unavailable in the proximal alimentary tract and that encapsulation technologies will likely be needed to deliver effective concentrations of these compounds to the lower gut to achieve in vivo reductions of Campylobacter .

Three experiments were conducted to evaluate the influence of vitamin D on fecal shedding of Escherichia coli O157:H7 in cattle. In the first experiment, two groups of cattle (beef and dairy) were assigned to a control treatment or to receive 0.5 × 10 6 IU vitamin D per day via oral bolus for 10 days. Fecal samples were collected before and throughout the dosing period for culture of E. coli O157:H7. No differences were observed for fecal shedding of E. coli O157:H7 among treatments for either beef or dairy animals. Serum concentrations of vitamin D were markedly higher ( P < 0.0001) in treated beef cattle but only tended to be higher ( P = 0.09) in the dairy cattle. In the second experiment, three successive vitamin D dosages (2,400, 4,800, and 9,600 IU/day; 14 days each) were administered to 14 dairy steers (7 steers served as controls), fecal samples were collected daily, and serum samples were collected weekly throughout the 42-day experimental period. No significant differences in fecal prevalence or serum vitamin D concentrations were observed for any of the vitamin D dosages. A third experiment sampled feedlot cattle (winter and summer) to determine whether serum vitamin D concentrations were correlated with fecal shedding of E. coli O157:H7. A fecal sample and a blood sample were obtained in each season from 60 randomly selected animals (total of 120 fecal samples and 120 corresponding blood samples). As expected, season was highly correlated ( r = 0.66) with serum vitamin D concentration with higher concentrations ( P < 0.01) observed in the summer. E. coli O157:H7 prevalence (percentage of positive samples) was not highly correlated ( r = 0.16) with season, although the correlation tended to be significant ( P = 0.08). The proportion of cattle shedding E. coli O157:H7 was 16.7 and 6.7% for the summer and winter collections, respectively. Results of this research do not support a correlation between vitamin D intake and E. coli O157:H7 shedding in cattle.

Foodborne pathogenic bacteria such as Escherichia coli O157:H7 are threats to the safety of beef. Citrus peel and dried orange pulp are by-products from citrus juice production that have natural antimicrobial effects and are often incorporated into least-cost ration formulations for beef and dairy cattle. This study was designed to determine if orange peel and pulp affected E. coli O157:H7 populations in vivo. Sheep ( n = 24) were fed a cracked corn grain–based diet that was supplemented with a 50-50 mixture of dried orange pellet and fresh orange peel to achieve a final concentration (dry matter basis, wt/wt) of 0, 5, or 10% pelleted orange peel (OP) for 10 days. Sheep were artificially inoculated with 10 10 CFU of E. coli O157:H7 by oral dosing. Fecal shedding of E. coli O157:H7 was measured daily for 5 days after inoculation, after which all animals were humanely euthanized. At 96 h postinoculation, E. coli O157:H7 shedding was reduced ( P < 0.05) in sheep fed 10% OP. Populations of inoculated E. coli O157:H7 were reduced by OP treatment throughout the gastrointestinal tract; however, this reduction reached significant levels in the rumen ( P < 0.05) of sheep fed 10% OP diets. Cecal and rectal populations of E. coli O157:H7 were reduced ( P <0.05) by inclusion of both 5 and 10% OP diets. Our results demonstrate that orange peel products can be used as a preharvest intervention strategy as part of an integrated pathogen reduction scheme.

Ractopamine HCl and zilpaterol HCl, β-agonists recently approved for use in feedlot cattle to improve performance traits and carcass leanness, were examined for their effects on fecal shedding of Escherichia coli O157:H7 in feedlot cattle. Fecal samples ( n = 2,454) were obtained from four experiments (one ractopamine HCl, three zilpaterol HCl) over the course of a 3-year period, either by rectal palpation (ractopamine HCl experiment) or from pen-floor fecal pats. Samples were cultured quantitatively and qualitatively for E. coli O157:H7. No significant treatment differences were detected for fecal prevalence of E. coli O157:H7 in the ractopamine HCl experiment. Zilpaterol HCl feeding had no effect ( P > 0.20) on fecal shedding in the first or second experiments, with overall E. coli O157:H7 prevalence relatively low (<7%). In the third zilpaterol HCl experiment, the percentage of fecal samples that were E. coli O157:H7 positive following qualitative culture was higher ( P < 0.05) in the zilpaterol HCl treatment (10.3%) than for the control (6.1%). The current research showed minimal effects of β-agonists on fecal shedding of E. coli O157:H7 and indicated that these compounds (fed immediately prior to slaughter) are not a cause for concern from a food safety standpoint.

The present study examined the incidence of Escherichia coli O157:H7 and Salmonella in feedlot lambs. Fifty-six feedlot lambs from eight sheep farming operations were grouped in a single drylot pen, fed, and managed as is typical in the southwestern United States. Fecal samples were collected on days 0, 46, 87, and 122 of the feeding period via rectal palpation. Wool samples (ventral midline) were collected one time only at the feedlot, immediately prior to shipping to the processing plant, and carcass swabs were collected following slaughter. All samples were cultured for E. coli O157:H7, Salmonella , and fecal coliforms, and select isolates were examined for antimicrobial susceptibility. Overall, the percentages of fecal and wool samples positive for E. coli O157:H7 averaged 9 and 18%, respectively. One carcass swab was E. coli O157:H7 positive. Of the 155 fecal samples collected, 11 (7%) were Salmonella positive. Salmonella was detected in nearly 50% of the wool samples collected prior to slaughter, while none of the carcasses were Salmonella positive 24 h postslaughter. All isolates ( E. coli O157:H7, Salmonella , and fecal coliforms) were susceptible to ceftiofur, enrofloxacin, and trimethoprim-sulfamethoxazole. One E. coli O157:H7 isolate cultured from a carcass swab was resistant to seven antibiotics, and seven wool E. coli O157: H7 isolates were multidrug resistant. Results of this research demonstrate that feedlot sheep are naturally colonized with E. coli O157:H7 and Salmonella and wool can be a source of carcass contamination; however, in-plant processing procedures and intervention strategies were largely effective in preventing carcass contamination.

The objectives of the current research were twofold: (i) to determine the prevalence of multidrug-resistant (MDR) Salmonella in the various classes of dairy cattle and (ii) to determine if comingling of calves from multiple farms at a heifer feedlot serves as a transmission vector for Salmonella back to the dairy farm. Four large commercial dairies in the southwestern United States were sampled in October 2005 and again in March 2006. Fecal samples were collected from hutch calves, 12-and 24-month-old heifers, lactating cows, dry cows, and cattle in the sick-fresh pen and cultured with brilliant green agar supplemented with novobiocin (BGA nov ) to estimate the overall Salmonella prevalence, or with tetracycline (BGA tet ) to estimate MDR Salmonella . Antimicrobial susceptibility testing was conducted with the National Antibiotic Resistance Monitoring System (NARMS) testing panel, and a portion of the isolates were serotyped. Salmonella prevalence among groups ranged from 0 to 96% positive, with the highest incidence observed in the hutch calves and cattle in the sick-fresh pen. Twenty-eight different serotypes were identified with serotype Reading accounting for the majority of isolates cultured on BGA tet . Nearly all (100 of 103) isolates cultured on BGA nov and screened for antibiotic resistance were pan susceptible, whereas over one-half (64%) of the isolates cultured on BGA tet were MDR. Forty isolates displayed the ACSSuT resistance pattern, and 36 isolates displayed the MDR-AmpC pattern of the 72 isolates examined following culture on BGA tet . The incidence of Salmonella cultured on BGA tet was low (9%) in all heifers and only one MDR isolate was cultured (from a 12-month-old heifer), suggesting the risk of transmission of Salmonella from the heifer feedlot back to the dairy is low. Results of this research suggest the incidence of MDR Salmonella , found primarily in hutch calves and cattle in the sick-fresh pen, is low in comparison to the overall Salmonella prevalence.

The effect of commercially available chestnut and mimosa tannins in vitro (experiment 1) or in vivo (experiment 2) on the growth or recovery of Escherichia coli O157:H7 or generic fecal E. coli was evaluated. In experiment 1, the mean growth rate of E. coli O157:H7, determined via the measurement of optical density at 600 nm during anaerobic culture in tryptic soy broth at 37°C, was reduced ( P < 0.05) with as little as 400 μg of either tannin extract per ml of culture fluid. The addition of 200, 400, 600, 800, and 1,200 μg of tannins per ml significantly ( P < 0.01) reduced the specific bacterial growth rate when compared with the nontannin control. The specific growth rate decreased with increasing dose levels up to 800 μg of tannins per ml. Bacterial growth inhibition effects in chestnut tannins were less pronounced than in mimosa tannins. Chestnut tannin extract addition ranged from 0 to 1,200 μg/ml, and a linear effect ( P < 0.05) was observed in cultures incubated for 6 h against the recovery of viable cells, determined via the plating of each strain onto MacConkey agar, of E. coli O157:H7 strains 933 and 86-24, but not against strain 6058. Similar tests with mimosa tannin extract showed a linear effect ( P < 0.05) against the recovery of E. coli O157:H7 strain 933 only. The bactericidal effect observed in cultures incubated for 24 h with the tannin preparations was similar, although it was less than that observed from cultures incubated for 6 h. When chestnut tannins (15 g of tannins per day) were infused intraruminally to steers fed a Bermuda grass hay diet in experiment 2, fecal E. coli shedding was lower on days 3 ( P < 0.03), 12 ( P = 0.08), and 15 ( P < 0.001) when compared with animals that were fed a similar diet without tannin supplementation. It was concluded that dietary levels and sources of tannins potentially reduce the shedding of E. coli from the gastrointestinal tract.

The effects of coincubating the active agent of an experimental chlorate product with nitrate or select nitro compounds, possible inducers and competing substrates for the targeted respiratory nitrate reductase, on concentrations of experimentally inoculated Salmonella enterica serovar Typhimurium and indigenous Escherichia coli were determined. Studies were completed in swine fecal suspensions as a prelude to the administration of these inhibitors to pigs. Results confirmed the bactericidal effect of chlorate (5 to 10 mM) against these fecal enterobacteria, reducing ( P < 0.05) concentrations by >2 log CFU ml −1 after 3 to 6 h of incubation. An effect ( P < 0.05) of pH was observed, with considerable regrowth of Salmonella and E. coli occurring after 24 h of incubation in suspensions buffered to pH 7.1 but not in suspensions buffered to pH 6.5 or 5.6. A 24-h coincubation of fecal suspensions with 5 to 10 mM chlorate and as little as 2.5 mM nitrate or 10 to 20 mM 2-nitro-1-propanol, 2-nitroethanol, and, sometimes, nitroethane decreased ( P < 0.05) Salmonella but not necessarily E. coli concentrations. 2-Nitro-1-propanol and 2-nitroethanol exhibited inhibitory activity against Salmonella and E. coli by an undetermined mechanism, even in the absence of added chlorate.

Dairy cows may serve as asymptomatic carriers of Salmonella . The potential for herd carrier status increases with herd size, and Salmonella shedding may be triggered by stresses placed on the animals. The scope of the current study is to determine the effects lactation may have on Salmonella genotypic diversity among detected serotypes. Fecal samples were collected on two sampling dates from 60 nonlactating and 60 lactating Holstein cows. No serotype was predominant over the two collection dates, although Salmonella Albany, Salmonella Anatum, Salmonella Newport, and Salmonella Senftenberg were detected in relatively high numbers. Twenty-three genotypes were detected on the first date and 27 on the second date. The greatest genotypic diversity was seen among Salmonella Newport and Salmonella Senftenberg, with five and nine genotypes, respectively. The presence of multiple serotypes and genotypes in the herd suggests multiple contamination sources. However, there was no conclusive effect of lactation status of the cows on Salmonella genotypic shedding.

The effect of 2-nitropropanol (2NPOH) administration on Salmonella enterica serovar Typhimurium in experimentally infected chicks was determined. Chicks orally challenged with 10 6 CFU/ml of a novobiocin- and naladixic acid–resistant Salmonella Typhimurium at 6 days of age were divided into three groups receiving 0 (control), 6.5, and 13 mg 2NPOH per bird (experiment 1) or four groups receiving 0 (control), 13, 65, and 130 mg 2NPOH per bird (experiment 2). Treatments were administered orally 1 day post– Salmonella challenge. Cecal contents collected at necropsy 24 and 48 h after treatment were subjected to bacterial and volatile fatty acid (VFA) analysis. In experiment 1, concentrations (mean ± SD log CFU per g) of Salmonella were reduced ( P < 0.05) in the group administered 13 mg 2NPOH per bird at both the 24- and 48-h samplings compared with the controls (2.58 ± 2.10 versus 4.64 ± 1.79 and 2.88 ± 2.78 versus 5.03 ± 2.42 at 24 and 48 h, respectively). In experiment 2, mean ± SD populations of Salmonella were reduced ( P < 0.05) in all groups receiving 2NPOH compared with untreated controls (3.65 ± 2.01, 3.39 ± 2.42, and 3.47 ± 1.55 at 13, 65, and 130 mg, respectively, versus 6.09 ± 1.02). Propionate concentrations were reduced ( P < 0.05) by the 13-mg 2NPOH per bird treatment. Total VFA concentrations from the group treated with 13 mg 2NPOH per bird were lower ( P < 0.05) by 48, but not 24, hours posttreatment than those from the group treated with 6.5 mg 2NPOH per bird. These results demonstrate the inhibitory activity of 2NPOH against Salmonella Typhimurium in vivo.

The objective of the present study was to determine the prevalence of intestinal Campylobacter in lactating dairy cows from various regions of the United States. Participating commercial dairy farms were chosen at random and were part of a national survey to determine E. coli O157:H7 and Salmonella prevalence in dairy cows. Farms had no previous history of Campylobacter problems. Fecal samples were collected rectally from 720 cows on farms in the northeast (four farms), in the desert southwest (three farms), and in the Pacific west (two farms). A minimum of 60 fecal samples per visit were collected from each farm. Thirty isolates were analyzed using the RiboPrinter Microbial Characterization System to obtain ribosomal RNA patterns. Twenty isolates were tentatively identified as Campylobacter jejuni , two as Campylobacter coli , three as Campylobacter spp., and five as unknown. Individual single-visit farm prevalence ranged from 0 to 10%. The disk diffusion method, employing 11 antibiotics, was used to test the antibiotic sensitivities of 27 of the isolates. Eight isolates were resistant to two or more antibiotics, 13 isolates were resistant to one antibiotic, and 6 were totally susceptible. Under the conditions of this study, the authors conclude that Campylobacter prevalence in lactating dairy cows in the United States is low, there is no difference in prevalence on the basis of geographical location, the predominant species is C. jejuni , and that the majority of these isolates are sensitive to antibiotics.

The effects of two feed supplements on Salmonella Typhimurium in the ceca of market-age broilers were determined. Broilers orally challenged 6 days before slaughter with a novobiocin- and nalidixic acid–resistant strain of Salmonella Typhimurium were divided into one of four groups (20 birds each). The first group (the control group) received no treatment, the second group received sodium nitrate (SN) treatment (574 mg of NaNO 3 per kg of feed), the third group received experimental chlorate product (ECP) treatment (15 mM NaClO 3 equivalents), and the fourth group received ECP treatment in combination with SN treatment. The SN treatment was administered via feed for 5 days immediately before slaughter, and ECP was provided via ad libitum access to drinking water for the last 2 days before slaughter. Cecal contents were subjected to bacterial analysis. Significant ( P < 0.05) Salmonella Typhimurium reductions (ca. 2 log units) relative to levels for untreated control broilers were observed for broilers receiving ECP in combination with SN. The ECP-only treatment resulted in significant ( P < 0.05) reductions (ca. 0.8 log) of Salmonella Typhimurium in trial 2. We hypothesize that increasing Salmonella Typhimurium nitrate reductase activity resulted in increased enzymatic reduction of chlorate to chlorite, with a concomitant decrease in cecal Salmonella Typhimurium levels. On the basis of these results, preadaptation with SN followed by ECP supplementation immediately preharvest could be a potential strategy for the reduction of Salmonella Typhimurium in broilers.

Salmonella cause economic losses to the swine industry due to disease and compromised food safety. Since the gut is a major reservoir for Salmonella , strategies are sought to reduce their concentration in pigs immediately before processing. Respiratory nitrate reductase activity possessed by Salmonella also catalyzes the intracellular reduction of chlorate (an analog of nitrate) to chlorite, which is lethal to the microbe. Since most gastrointestinal anaerobes lack respiratory nitrate reductase, we conducted a study to determine if chlorate may selectively kill Salmonella within the pig gut. Weaned pigs orally infected with 8 × 10 7 CFU of a novobiocin- and nalidixic acid–resistant strain of Salmonella Typhimurium were treated 8 and 16 h later via oral gavage (10 ml) with 0 or 100 mM sodium chlorate. Pigs were euthanized at 8-h intervals after receiving the last treatment. Samples collected by necropsy were cultured qualitatively and quantitatively for Salmonella and for most probable numbers of total culturable anaerobes. A significant ( P < 0.05) chlorate treatment effect was observed on cecal concentrations of Salmonella , with the largest reductions occurring 16 h after receiving the last chlorate treatment. An observed treatment by time after treatment interaction suggests the chlorate effect was concentration dependent. Chlorate treatment may provide a means to reduce foodborne pathogens immediately before harvest.