Search

Two experiments were conducted to evaluate the feeding of direct fed microbials (DFMs) on fecal shedding of Escherichia coli O157:H7 in naturally infected cattle (experiment I) and on Salmonella in the feces and peripheral lymph nodes (PLNs) of experimentally infected cattle (experiment II). Thirty cattle, 10 per treatment, were used in each experiment. Treatments in experiment I consisted of a control (lactose carrier only); DFM1, a 1:1 ratio of Enterococcus faecium and Lactobacillus animalis; and DFM2, a 1:1 ratio of Lactobacillus acidophilus and Pediococcus acidilactici. In Experiment II, DFM1 was replaced with DFM3, a 1:2 ratio of Lactobacillus reuteri and other Lactobacillus strains. Additives were mixed in water and applied as a top-dressing to each pen's daily ration for 50 days. Approximately half-way through each experiment, the DFM concentration was doubled for the remainder of the study. Fecal samples were collected throughout experiment I and cultured for E. coli O157:H7. Cattle in experiment II were inoculated intradermally with Salmonella Montevideo on days 32, 37, and 42 and then necropsied on days 49 and 50 (five cattle per treatment on each day). Innate immune function was assessed on days 29, 49, and 50. In experiment I, fecal concentration and prevalence of E. coli O157:H7 were not different (P > 0.10) nor was there an effect (P = 0.95) on the percentage of super shedders (cattle shedding ≥3.0 log CFU/g of feces). In experiment II, no treatment differences (P > 0.05) were observed for Salmonella in the PLNs except for the inguinal nodes, which had a significantly lower Salmonella prevalence in DFM-supplemented cattle than in the controls. Immune function, as measured by monocyte nitric oxide production and neutrophil oxidative burst, was decreased (P < 0.05) in the DFM treatment groups. Although results of this research indicate little to no effect of these DFMs on E. coli O157:H7 or Salmonella in cattle, an increase in the duration of administration to that similar to what is used for commercial cattle might elicit treatment differences.

A series of proof-of-concept studies were developed to determine whether a commercial bacteriophage cocktail could be utilized for the mitigation of Salmonella in bovine peripheral lymph nodes (LNs). The first objective sought to determine whether exogenous phage could be isolated from the LNs following administration. If isolation were successful, the second objective was to determine whether the phage in the LNs could effectively reduce Salmonella. Salmonella Montevideo was inoculated intradermally at multiple sites and multiple times, followed by delivery of the phage cocktail subcutaneously in two injections around each of the right and left prescapular and subiliac LNs. At the conclusion of each study, animals were euthanized, and the popliteal, prescapular, and subiliac LNs were examined. The inoculated phage was successfully isolated from the LNs; transmission electron microscopy revealed phages in the LNs of the treated cattle, and these phages were identical to those in the cocktail. Levels of phage were higher (P0.01) in the prescapular and subiliac LNs in the phage-treated than in the control cattle. In subsequent studies, the protocols were modified to increase Salmonella and phage levels within the LNs. Compared with the first study, overall Salmonella levels were increased in the LNs, and phage treatment decreased (P0.01) Salmonella in the some of the LNs. Phage levels were numerically but not significantly increased (P = 0.12) in the treated cattle. The final study was modified, hypothesizing that a 48-h postmortem period before LN removal would facilitate phage-Salmonella interaction; however, no differences (P0.10) in Salmonella levels were found among treatments. Salmonella-specific phages administered to live cattle can translocate to the LNs; however, these phages had limited to no effect on Salmonella in these LNs under these experimental conditions.

Two experiments were conducted to evaluate the feeding of direct fed microbials (DFMs) on fecal shedding of

Broiler chicks were inoculated by gavage on the day of hatch with a characterized continuous-flow (CF3) competitive-exclusion culture that contained 29 different bacterial isolates to determine the effects on Salmonella cecal and crop colonization during grow-out. Chicks at 3 days old were challenged by gavage with 104 Salmonella typhimurium . Propionic acid significantly increased (P ≤ 0.001) in the ceca of 3-day-old CF3-treated chicks compared to control chicks. Ceca from market-age control chickens in two trials contained log 2.6 and log 1.4 Salmonella CFU/g of cecal contents, respectively, while log 0.4 Salmonella CFU/g of cecal contents were detected in both trials in ceca from CF3-treated chickens. Percentages of Salmonella culture-positive ceca in the two trials, respectively, were 80% and 60% in controls and 27% in treated chickens in both trials. Crops from market-age control chickens in the two trials averaged log 0.7 Salmonella CFU/g of cecal contents, while crops from treated chickens averaged log 0.4 CFU/g of cecal contents. In trial 1, 60% of control chick crops and 27% of treated chick crops tested Salmonella culture positive. Litter contamination by Salmonella spp. at 5 weeks was reduced significantly (P ≤ 0.01) in pens of CF3-treated groups compared to litter from control pens. Results indicate that CF3 reduced cecal and crop colonization by S. typhimurium during grow-out, which may reduce the number of Salmonella cells entering the processing plant and decrease the potential for carcass contamination during processing.

A rumen fluid-based differential carbohydrate agar medium for enumerating chicken cecal carbohydrate-utilizing bacteria was tested. Eleven bacteria isolated from a continuous-flow culture that had been seeded with chicken cecal contents were inoculated on fifteen different media in an anaerobic environment. These media included various levels of rumen fluid (0%, 5%, 10%, 16%) and carbohydrate (glucose [GLU], galactose [GAL], lactose[LAC]). Viable cell recovery (colony-forming units [CFU]/direct cell counts × 100%) was higher for 16% rumen fluid-supplemented carbohydrate media than the lower concentrations of rumen fluid-supplemented carbohydrate media. In addition, average viable cell recovery of all 11 bacteria on differential carbohydrate media were significantly (P < 0.05) higher in galactose and lactose media than on the glucose media. Data indicated that 16% rumen-fluid based media can be used to enumerate anaerobic, lactose-fermenting subgroups of chicken cecal bacteria.

Bacterial colonization of cecal mucosal epithelium in 3-day-old chicks administered a characterized continuous-flow (CF) culture of 29 microorganisms on the day of hatch was evaluated by scanning electron microscopy. Extensive colonization of the mucosa was noted in the ceca of CF-treated chicks, with large colonies of bacteria located predominately within and between crypts. Cecal crypts from control chicks contained only thin strands of mucus with a few bacteria. Individual cells and clumps of bacteria were observed bound to the mucosal epithelium in both CF-treated and control chicks. Colonization by CF culture bacteria was accompanied by an increase in the concentration of volatile fatty acids in the cecal contents and increased resistance to colonization by Salmonella typhimurium .

The effect of oral inoculation with a defined culture of cecal bacteria and provision of dietary lactose on colonization resistance to invasive Salmonella enteritidis was evaluated in leghorn chicks. A defined mixed culture composed of 11 indigenous cecal bacteria capable of utilizing lactose or lactose fermentation products as a primary carbon source was isolated and maintained in continuous-flow culture. Combined treatment with the defined culture and dietary lactose significantly decreased (P < 0.01) the number of Salmonella in the cecal contents and the number of Salmonella cecal-culture-positive chicks in four replicated trials. Additionally, Salmonella colonization of the spleen, liver, and cecal tonsils was significantly decreased (P < 0.05) in each of the trials. The results indicate that the resistance of leghorn chicks to S. enteritidis cecal and organ colonization is effectively increased by inoculation with a defined culture of cecal bacteria and provision of lactose in the diet.

Biting arthropods are implicated in the transdermal transmission of Salmonella to bovine peripheral lymph nodes, and such contamination can contribute to increased Salmonella prevalence in processed beef. Since horn flies can acquire Salmonella and then excrete the bacteria in their feces, on-animal fly infestations were conducted in this study to assess whether horn flies have a role in this bacterial transmission. Three Salmonella serotypes were used to assess fly acquisition from and excretion onto cattle. The results indicated that flies can acquire Salmonella from the hide, as assessed by recovery from homogenates of surfacesterilized flies, and that Salmonella persists for at least 5 days in the fly. Fly fecal excreta serves as a bacterial contaminant on the hide, and the overall mean probable estimate of the quantity shed was ≈10(5) most probable number per fly cage area. In 5 days, no transmission of the bacteria to bovine peripheral lymph nodes was evident, prompting an assessment of the effects of prolonged horn fly feeding on transmission. Three groups of animals were infested with flies that had consumed a blood meal containing Salmonella Senftenberg. After 5 days, the study was either terminated or the flies were removed and the cages replenished with unfed flies either once or twice over the course of an 11- or 19-day fly exposure period, respectively. A microlancet-inoculated positive-control animal was included in each group for comparison. The impact of prolonged horn fly feeding was evident, as 8% of lymph nodes cultured were positive from the 5-day exposure, whereas 50 and 42% were positive from 11- and 19-day exposures, respectively. Higher concentrations of Salmonella were recovered from fly-infested animals than from the microlancet-inoculated control, likely a result of repeated inoculations over time by flies versus a single introduction. The data described provide new insights into the transmission dynamics of Salmonella in cattle populations, highlighting a role for biting flies as an important reservoir.

Previous attempts to infect peripheral lymph nodes (PLNs) with Salmonella via oral inoculation have been inconsistent. Therefore, we performed a series of experiments to determine whether multiple exposures to an oral challenge would result in Salmonella-positive PLN in cattle. In each of three experiments, calves were inoculated with Salmonella Montevideo. In the first experiment, calves were challenged with either no Salmonella (control), a single oral dose (~1010; PCON), or 10 consecutive doses in water (~103; WAT). The positive control treatment resulted in an increase (P < 0.05) in the percentage of Salmonella-positive PLNs, compared with the WAT-treated and control animals. Experiments 2 and 3 were designed to additionally determine if the stress associated with feed and water deprivation influences the systemic spread of Salmonella from the gastrointestinal tract to PLNs. Following 14 days of oral inoculation (average 7.1 × 104 CFU/day) in experiment 1, Salmonella was recovered from one subiliac an...

Because challenge models to infect peripheral lymph nodes (PLNs) with Salmonella have not been reported, we performed a series of experiments to develop and refine challenge models to evaluate an intervention applied at the animal level and to provide initial estimates of efficacy of an intervention (i.e., a vaccine) to aid in the design of future studies. In each of four experiments, steers (control or vaccinated) were inoculated with Salmonella strains Montevideo or Newport, and in experiment IV, Salmonella Senftenberg was also used. Calves were euthanized 14 to 42 days postinoculation, and PLNs were collected. In the first experiment, calves were challenged with ∼10¹⁰ Salmonella cells, and few treatment differences were observed 14 days postchallenge. However, by day 21, Salmonella Newport was recovered from fewer vaccinated calves than control calves (P0.05). In experiment II, calves were challenged with ∼10⁷ Salmonella cells and, after two necropsies (14 and 28 days postchallenge), only one lymph node was Salmonella positive; therefore, the study was terminated. In experiment III, calves were again challenged with ∼10¹⁰ Salmonella cells, and no significant effect of vaccine was observed in calves challenged with Montevideo or Newport strains. A transdermal route of challenge was explored in experiment IV, using a 10-lancet, allergy testing instrument. Sixteen steers were challenged with either Salmonella Newport or Salmonella Montevideo (Salmonella Newport right legs; Salmonella Montevideo left legs), and all steers were challenged on the lower abdomen with Salmonella Senftenberg. Transdermal inoculation resulted in predictably Salmonella-positive PLNs, and a modest vaccine effect was detected. Because it is well tolerated by the calves and results in predictable and regionally specific Salmonella recovery from PLNs, the transdermal route of challenge may be preferred by researchers wishing to evaluate the impact of interventions designed to reduce the carriage of Salmonella in PLNs.

Recent investigations have found that Salmonella can be routinely recovered from peripheral lymph nodes (PLNs) of cattle presented for harvest. When contained within the PLNs, this foodborne pathogen is protected from currently used postharvest, inplant intervention strategies and, therefore, PLNs harboring Salmonella may be a potential contaminant of ground beef. The objective of this work was to develop a challenge model that effectively and repeatedly results in Salmonella -positive PLNs. A 10-lancet skin-allergy instrument was inoculated with Salmonella, and calves were inoculated intra- and/or transdermally by applying the device over various ventral regions of the skin. Salmonella was successfully and predictably recovered from regionspecific PLNs up to 8 days postchallenge. Furthermore, serotypes inoculated within specific regions were only recovered from the PLNs draining those regions. This model provides a method to predictably infect PLNs with Salmonella. Further, this model makes it possible to determine the duration of infection and to evaluate candidate interventions that may shorten the duration of infection.

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 O45O145O121O111O26O103 and among human strains from serogroups O145O103O26O111O121O45. 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 DCR 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.

Utilizing a transdermal method of inoculation developed in our laboratory, the duration of infection of Salmonella in the peripheral lymph nodes of steers was examined. Thirty-six Holstein steers (mean body weight of 137 kg) were inoculated with Salmonella Montevideo (day 0) on each lower leg and both sides of the back and abdomen. Calves were euthanized beginning at 6 h and subsequently on each of days 1, 2, 4, 7, 9, 11, 14, and 21 postinoculation (four animals each time). The subiliac, popliteal, and superficial cervical (prescapular) lymph nodes were collected and cultured (quantitatively and qualitatively) for the challenge strain of Salmonella. The challenge strain was detected via direct culture within the lymph nodes at 6 h postinoculation and on each subsequent necropsy date. Salmonella levels in lymph node were 0.8 to 1.8 log CFU/g. Lymph nodes were generally positive after enrichment culture throughout the experiment. Salmonella elimination appeared to begin approximately 14 days postinoculation. However, elimination was not completed by day 21; therefore, a second experiment was conducted identical to the first except that the time from inoculation to necropsy was extended. Salmonella was recovered via direct culture on each of the necropsy days, and results in general were similar to those of experiment I, except that on days 20, 24, and 28 isolates from serogroups C2 and E1 were identified in addition to the inoculation strain C1 in multiple animals. The data from both experiments indicate that after a single inoculation event, Salmonella would be completely cleared by approximately 28 days. Further research with expanded times between inoculation and necropsy is required for verification.

Although thymol is bactericidal against many pathogens in vitro, its in vivo effectiveness against pathogens in the lower gastrointestinal tract is limited because of its rapid absorption in the proximal gut. Thymol-β-smallD/small-glucopyranoside (β-thymol), a conjugated form of thymol, can deliver thymol to the lower gastrointestinal tract and has shown antibacterial effects. In the present study, we examined the in vitro effects of β-thymol on Salmonella enterica serovar Typhimurium (ST) and Escherichia coli K88 (K88). We inoculated one-half strength Mueller-Hinton broth with 5.8 ± 0.09 log CFU/ml novobiocin- and naladixic acid-resistant (NN) ST (NVSL 95-1776) and 5.1 ± 0.09 log CFU ml(-1) NN-resistant K88, with or without porcine feces (0.1% [wt/vol]) (fecal incubations). The resultant bacterial suspensions were distributed under N2 to triplicate sets of tubes to achieve initial concentrations of 0, 3, 6, and 12 mM for ST treatments and 0, 3, 12, and 30 mM for K88 treatments. Samples were incubated at 39°C and then plated onto NN-containing brilliant green agar and NN-containing MacConkey agar; ST and K88 CFU concentrations were determined via 10-fold dilutions, and viable cell counts were performed at 0, 6, and 24 h. No differences in ST CFU counts were observed in β-thymol-treated tubes without the added porcine feces (i.e., pure culture) at 6 or 24 h. However, in tubes that contained fecal incubations, ST CFU counts were reduced (P0.05) from controls at 6 h in tubes treated with 6 and 12 mM β-thymol, whereas in tubes treated with 3, 6, and 12 mM β-thymol the CFU counts were reduced (P0.05) at 24 h. No differences were observed in K88 CFU counts in pure culture or in fecal incubations at 6 h, but K88 CFU counts were reduced (P0.05) in both pure and fecal incubations at 24 h. The results from this study demonstrate that β-thymol, in the presence of fecal suspensions, has anti-Salmonella and anti-E. coli effects, suggesting a role of β-glycoside-hydrolyzing microbes for the release of bactericidal thymol from β-thymol.

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 (P0.0001) 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 (P0.01) 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.

Strategies aimed at reducing fecal shedding of Salmonella and other foodborne pathogens may be effective for limiting transmission of pathogens from food animals to humans. The objective of this study was to determine the effectiveness of gallium maltolate (GaM) against Salmonella in vitro and to determine whether oral administration of GaM would reduce fecal shedding of Salmonella in cattle. Gallium is a semimetal exhibiting antimicrobial properties against some pathogenic bacteria, including Salmonella, by exploiting their need for iron to survive and replicate. In vitro growth studies were performed in pure cultures of Salmonella and in mixed cultures from ruminal fluid. Inclusion of GaM in culture medium or in mixed cultures of ruminal fluid resulted in a significant reduction in growth of Salmonella, suggesting that GaM may be effective for limiting growth and survival in vivo. Therefore, we subsequently administered two doses of GaM to Holstein steers, experimentally infected them with Salmonella, and quantitatively and qualitatively monitored fecal shedding at 12-h intervals. Sixty hours after beginning treatment, cattle were euthanized, and luminal contents and tissue were aseptically harvested from the rumen, jejunum, spiral colon, cecum, and rectum. The luminal contents were processed for quantitative and qualitative analysis of the challenge strains of Salmonella, and tissue samples were enriched and plated for qualitative analysis. We found no significant differences between control and treated animals in quantitative levels of Salmonella in the feces or the luminal contents. Likewise, we observed no pattern between control and treated animals in the frequency of positive or negative results from enriched feces, luminal contents, or tissue samples. These results suggest that GaM was not effective for reducing Salmonella in cattle.

Recent concerns about the use of antimicrobials in food animals have increased interest in the microbial ecology and biofilms within their gastrointestinal tract. This work used a continuous-flow chemostat system to model the microbial community within the ceca from day-of-hatch chicks and its ability to resist colonization by Salmonella enterica serovar Typhimurium. We characterized the biofilm and planktonic communities from five cultures by using automated ribotyping. Eight species from six different genera were identified. Overall, the planktonic communities were more diverse, with 40% of the cultures containing four or more bacterial species. Eighty percent of the biofilm communities contained only one or two species of bacteria. Enterococcus faecalis was the only species isolated from all communities. None of the resulting microbial communities was able to resist colonization by S. enterica serovar Typhimurium. This is the first study to provide a molecular-based characterization of the biofilm and planktonic communities found in day-of-hatch chicken cecal microflora cultures.

This study evaluated the composition of gastrointestinal bacterial communities in birds during an age in which their susceptibility to Salmonella is highly diminished. One of the challenges to developing probiotics is to develop an efficacious culture of minimal diversity that includes bacteria that are vital contributors to protection from pathogens, but excludes unnecessary species. This study used in vitro continuous-flow culture techniques to test the ability of mixed bacterial cultures acquired from in vivo sources, to resist colonization by a marker Salmonella enterica serovar Typhimurium, and then characterized the constituents of both biofilm and planktonic communities by biochemical, phenotypic, and molecular methods. These cultures, initiated from 14-day-old chicks, were all able to restrict colonization by Salmonella in an average of 10 days. Eighteen species of bacteria from 10 different genera were characterized. However, each culture contained a mixture of only 11 species, which included lactic acid bacteria. Biofilms contained less than 50% of the species found in the planktonic communities. Although not adults, the diversity of microbes within the cecal cultures from 14-day-old birds represents a community complex enough to oppose colonization by a nonindigenous bacteria in vitro. These results describe bacterial mixtures containing less diversity than in previously described avian protective cultures.

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 (P0.05) concentrations by2 log CFU ml(-1) after 3 to 6 h of incubation. An effect (P0.05) 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 (P0.05) 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.

Previous attempts to infect peripheral lymph nodes (PLNs) with Salmonella via oral inoculation have been inconsistent. Therefore, we performed a series of experiments to determine whether multiple exposures to an oral challenge would result in Salmonella-positive PLN in cattle. In each of three experiments, calves were inoculated with Salmonella Montevideo. In the first experiment, calves were challenged with either no Salmonella (control), a single oral dose (∼10(10); PCON), or 10 consecutive doses in water (∼10(3); WAT). The positive control treatment resulted in an increase (P0.05) in the percentage of Salmonella-positive PLNs, compared with the WAT-treated and control animals. Experiments 2 and 3 were designed to additionally determine if the stress associated with feed and water deprivation influences the systemic spread of Salmonella from the gastrointestinal tract to PLNs. Following 14 days of oral inoculation (average 7.1 × 10(4) CFU/day) in experiment 1, Salmonella was recovered from one subiliac and one superficial cervical lymph node of calves that were deprived of feed and water (72 h). No treatment differences (P0.05) were observed between control and deprived calves. Based on the poor recovery of Salmonella from the PLNs in WAT-challenged calves in experiments 1 and 2, a higher challenge dose (average 1.2 × 10(7) CFU) was used in experiment 3. The increased dose resulted in the recovery of the challenge strain of Salmonella from the PLNs (70.8 and 75.0% of control and deprived calves, respectively). However, no treatment differences (P0.05) were observed between control and deprived calves. Results of this research demonstrated that a substantial oral challenge is required to produce Salmonella-positive PLNs. However, as the challenge periods examined herein were considerably shorter compared with the normal time spent by cattle in feedlots, increased exposure time to lower doses may produce the same effect observed in experiment 3.

Escherichia coli O157:H7 is a virulent foodborne pathogen that causes severe human illness and inhabits the intestinal tract of food animals. Colicins are antimicrobial proteins produced by E. coli strains that inhibit or kill other E. coli. In the present study, the efficacy of three pore-forming colicins (E1, N, and A) were quantified in vitro against E. coli O157:H7 strains 86-24 and 933. Colicins E1 and N reduced the growth of E. coli O157:H7 strains, but the efficacy of each colicin varied among strains. Colicin E1 was more effective against both strains of E. coli O157:H7 than colicins A and N and reduced (P < 0.05) populations of E. coli O157:H7 at concentrations

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 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 NaNO3 per kg of feed), the third group received experimental chlorate product (ECP) treatment (15 mM NaClO3 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.

Ruminant animals are a natural reservoir of the foodborne pathogen Escherichia coli O157:H7. Some foodborne pathogens (e.g., E. coli) are equipped with a nitrate reductase that cometabolically reduces chlorate. The intracellular reduction of chlorate to chlorite kills nitrate reductase-positive bacteria; however, species that do not reduce nitrate are not affected by chlorate. Therefore, it has been suggested that ruminants be supplemented with chlorate prior to shipment for slaughter in order to reduce foodborne illnesses in human consumers. Sheep (n = 14) were fed a high-grain ration and were experimentally infected with E. coli O157:H7. These sheep were given an experimental product (XCP) containing the equivalent of either 2.5 mM NaNO 3 and 100 mM NaCl (control sheep; n = 7) or 2.5 mM NaNO 3 and 100 mM NaClO 3 (chlorate [XCP]-treated sheep; n = 7). Control and XCP-treated sheep were treated for 24 h; XCP treatment reduced the population of inoculated E. coli O157:H7 (P < 0.05) from 10 2 , 105, and 10 5 CFU/g in the rumen, cecum, and rectum, respectively, to

The MICs of streptomycin for Salmonella isolates from swine and poultry were determined by a micro-broth dilution technique. The Salmonella isolates were recovered from the lymph nodes and cecal contents of market-age swine and from the cecal contents of poultry at the time of slaughter and were found by disk diffusion to be resistant to 10 μg of streptomycin. MIC testing was carried out with the Sensititre susceptibility system for streptomycin, which uses a microwell concentration gradient of 16 to 800 μg/ml. Results indicated that >80% of the swine isolates had MICs of ≤64 μg/ml, while 51% of poultry isolates exhibited MICs of ≥128 μg/ml. The highest MICs observed in swine and poultry were 256 and 800 μg/ml, respectively. Replicate tests performed on 12 of the isolates chosen at random indicated a 100% correlation between runs. Advantages of this system include easily read results and precoated wells. Disadvantages include the cost and the inability to test concentrations of streptomycin other than those in the wells. We found this micro-broth dilution commercial test kit to provide a relatively quick and easy testing procedure for the determination of streptomycin resistance in Salmonella. Cette etude determine les concentrations minimales inhibitrices (MICs) de streptomycine pour des isolats de Salmonella, resistantes a la streptomycine, sur de la viande de porc et de volaille par une technique de microdilution. Les resultats indiquent que plus de 80% des isolats de porc contiennent des teneurs en MICs inferieures a 64μg/ml tandis que 51% des isolats de volaille sont superieurs a 128μg/ml. Les resultats demontrent que cette methode est rapide et facile pour la determination de resistance a la streptomycine dans les Salmonella.

Campylobacter bacteria are foodborne pathogens that can colonize the gut of food animals. Limited in their ability to ferment sugars, Campylobacter can derive energy for growth via amino acid catabolism. The objectives of the present studies were to test whether supplemental distillers grains containing high amounts of rumen-undegradable intake protein or supplemental lasalocid may, by promoting amino acid flow to the lower bovine gut, increase intestinal carriage of Campylobacter. In study one, 10 steers (5 per treatment) were adapted to diets formulated to achieve 0 or 30% dried distillers grains. After an initial 14-day adaptation to the basal diet, control and treated steers were fed their respective diets for 23 days, after which time they were fed supplemental lasalocid for an additional 8 days, followed by a 5-day withdrawal. In study two, 24 steers preacclimated to a basal diet were adapted via 3-day periodic increases to dietary treatments formulated to achieve 0, 30, or 60% wet corn distillers grains with solubles. Analysis of Campylobacter bacteria cultured from duodenal and fecal samples in study one and from fecal samples in study two revealed no effect of dried distillers grains or wet corn distillers grains with solubles on the prevalence or concentrations of duodenal or fecal Campylobacter. The results from study one indicated that colonized steers, regardless of treatment, harbored higher Campylobacter concentrations when transitioned to the basal diet than when coming off pasture. Campylobacter carriage was unaffected by lasalocid. These results provide no evidence that feeding distillers grains high in rumen-undegradable intake protein or supplemental lasalocid contributes to increased intestinal carriage of Campylobacter in fed cattle.

Lymphokine (ILK) secreted from concanavalin A-stimulated T cells from Salmonella Enteritidis-immune chickens is an undefined mixture of proteins that confers protection against Salmonella infectivity when administered to day-old chicks. It has previously been shown that polyclonal antibodies raised against human granulocyte colony-stimulating factor (GCSF) can neutralize the heterophil activation that is responsible for ILK's protective effect. Western blot analysis of ILK probed with anti-GCSF antibodies detects a prominent protein of mass 33 kDa. We have sequenced the first 20 amino acids of this protein and found it to be identical to residues 24 to 43 of P33, a 326-amino acid protein of unknown function encoded by the chicken mim-1 gene. The primary structure of P33 consists of two 140-residue imperfect repeats that are each homologous to a mammalian neutrophil chemotactic factor termed leukocyte cell-derived chemotaxin 2 (LECT2). We have expressed mim-1 in Escherichia coli and demonstrated in vitro that recombinant P33 is chemotactic for heterophils, the avian equivalent of mammalian neutrophils. We have also constructed a derivative of P33 that consists of residues 33 to 165 (P33[33-165]), the first repeat sequence of P33 that is homologous to LECT2. P33(33-165) is chemotactic for heterophils both in vitro and in vivo, inducing an influx of heterophils into the peritoneum in a response similar to that observed with ILK. These results suggest that P33 functions as a chemotactic factor in chickens and that it plays an active role in ILK-mediated protection against Salmonella infection.

Antimicrobial resistance levels were examined for 365 Salmonella isolates recovered from the lymph nodes (n = 224) and cecal contents (n = 141) of market-age swine at slaughter. Antimicrobial resistance testing was performed by disk diffusion using 13 antibiotics common in the treatment of disease in human and veterinary medicine. Although none of the antibiotics tested were used subtherapeutically within the last 5 years on the farms sampled, resistance to chlortetracycline, penicillin G, streptomycin, and sulfisoxazole was common. Penicillin G resistance was significantly more frequent (P = 0.03) and sulfisoxazole resistance was significantly less frequent (P < 0.01) in lymph node versus cecal isolates. Multidrug resistance was observed among 94.7% of the lymph node isolates and 93.5% of the cecal isolates. The most frequent multidrug resistance pattern included three antibiotics-penicillin G, streptomycin, and chlortetracycline. Isolates in somatic serogroup B, and more specifically, Salmonella Agona and Salmonella Schwarzengrund isolates, were often resistant to a greater number of antibiotics than were isolates in the other serogroups. Streptomycin, sulfisoxazole, ampicillin (lymph node isolates), and nitrofurantoin (cecal isolates) resistance levels differed significantly between somatic serogroups. The prevalence of penicillin G-, streptomycin-, and sulfisoxazole-resistant isolates differed significantly between serovars for both lymph node and cecal isolates. Results of this study suggest that a correlation exists between the somatic serogroup or serovar of a Salmonella isolate and its antimicrobial resistance status, which is specific to the antibiotic of interest and the source of the isolate (lymph node versus cecal contents).

Escherichia coli O157:H7 and Salmonella Typhimurium DT104 are important foodborne pathogens affecting the beef and dairy industries and strategies are sought to rid these organisms from cattle at slaughter. Both pathogens possess respiratory nitrate reductase that also reduces chlorate to the lethal chlorite ion. Because most anaerobes lack respiratory nitrate reductase, we hypothesized that chlorate may selectively kill E. coli O157:H7 and Salmonella Typhimurium DT104 but not potentially beneficial anaerobes. In support of this hypothesis, we found that concentrations of E. coli O157:H7 and Salmonella Typhimurium DT104 were reduced from approximately 1,000,000 colony forming units (CFU) to below our level of detection (< or = 10 CFU) following in vitro incubation (24 h) in buffered ruminal contents (pH 6.8) containing 5 mM added chlorate. In contrast, chlorate had little effect on the most probable number (mean +/- SD) of total culturable anaerobes (ranging from 9.9 +/- 0.72 to 10.7 +/- 0.01 log10 cells/ml). Thus, chlorate was bactericidal to E. coli O157:H7 and Salmonella Typhimurium DT104 but not to potentially beneficial bacteria. The bactericidal effect of chlorate was concentration dependent (less at 1.25 mM) and markedly affected by pH (more bactericidal at pH 6.8 than pH 5.6).

Broiler chicks were spray treated on the day of hatch with titrated dosages (10(6), 10(7), or 10(8) anaerobic CFU) of a characterized competitive exclusion culture (CF3) and challenged orally on day 3 with 10(4) CFU of Salmonella typhimurium. On day 10, cecal contents from control and CF3-treated chicks were cultured for S. typhimurium to determine the minimal efficacious dosage of the CF3 culture. The experiment was repeated in three replicated trials. Resistance to Salmonella cecal colonization was dosage related and progressively enhanced at the 10(7)- and 10(8)-CFU dosages compared with the 10(6)-CFU dosage. The 10(7)-CFU dosage was selected as the minimal effective dosage and evaluated for efficacy during a 43-day broiler growout study. Six hundred broilers were spray treated on the day of hatch and compared with 600 controls. One-half of the control and CF3-treated birds were challenged orally on day 3 with 10(4) CFU of S. typhimurium and designated "seeders." The remaining unchallenged birds were designated "contacts." Compared with the controls, the recovery of Salmonella cells from the ceca of the CF3-treated broilers was significantly decreased (P0.01) in the challenged seeders on days 21 and 43 of growout. Salmonella contamination of floor pen litter was significantly reduced (P0.05) in pens of CF3-treated birds compared with controls. The transmission of Salmonella cells from seeder to contact birds in the same pens was decreased significantly (P0.01). The results indicated that treatment of broiler chicks on the day of hatch with the 10(7)-CFU dosage of CF3 culture effectively increased resistance to S. typhimurium challenge during growout to market age.

Older leghorn hens, more than 50 weeks of age, were divided into three groups designated 1, unmolted controls; 2, molted; or 3, molted treated with lactose. Forced molt was induced by 14 days of feed removal. Lactose was provided to the hens in group 3 as 2.5% (wt/vol) of the daily drinking water. Each hen in all groups was challenged orally with 10 5 Salmonella enteritidis (SE) cells on day 7 of feed removal. The study was repeated in three replicated trials. The concentrations of acetic, propionic, and total volatile fatty acids (VFA) in the cecal contents of the molted hens in groups 2 and 3 decreased significantly (P < 0.05) on days 6 and 14 of molt compared with the unmolted controls. Forced molt had no apparent effect on pH or on the oxidation-reduction potential of the ceca. Compared to the unmolted controls, SE cecal and spleen and liver colonization was significantly increased (P < 0.05) in the molted hens in group 2. Compared to the molted hens in group 2, SE cecal and spleen and liver colonization was significantly decreased (P < 0.05) in two of three trials in the hens in group 3 provided with lactose. The results suggested that the increased susceptibility of molting hens to SE colonization may be associated with decreased fermentation and production of VFA by cecal bacteria or by a depletion of the number of VFA-producing bacteria present in the ceca. The results further suggest that providing lactose in the drinking water during molting may significantly enhance resistance to SE colonization.

The disinfectant and antibiotic susceptibility profiles of 344 Escherichia coli O157:H7 strains from cattle carcasses, feces, and hides and ground beef from the United States were determined. A low prevalence of antibiotic resistance was observed (14%). The highest prevalences of resistance were to sulfisoxazole (10.5%), tetracycline (9.9%), streptomycin (7%), and chloramphenicol (4.9%). Four strains were resistant to eight antibiotics (two strains from ground beef and one strain each from hide and preevisceration carcass swabs of cull cattle at harvest). Pulsed-field gel electrophoresis analysis of the E. coli O157:H7 strains revealed two major groups (designated 1 and 2) composed of 17 and 20 clusters, respectively. Clusters 1A, 1B, 1C, and 1G.1 were associated with multidrug-resistant strains. There was no observed correlation between disinfectant resistance and antibiotic resistance. Sixty-nine (20%) of the 344 strains were resistant to chlorhexidine or benzalkonium chloride or the MICs of benzyldimethyldodecylammonium chloride were elevated. Inducible resistance was observed at elevated concentrations of antibiotics (1.4%) and disinfectants (6.1%). The highest rate of disinfectant inducible resistance was to OdoBan, quaternary ammonium chlorides, and the surface disinfectants F25, FS512, and MG, which are used in dairies, restaurants, and food processing plants. High MICs (1,024 to 4,096 m g/ml) of acetic, lactic, and citric acids were found. The decreasing order of acid potency based on molar MICs (MICs(molar)) was acetic, citric, and lactic acid. The correlation of the concentration of dissociated organic acids and MICs(molar) strongly suggests that the observed inhibition of E. coli O157:H7 was primarily due to dissociated forms of the acids.

A competitive-exclusion culture was isolated by the continuous-flow culture of chicken cecal bacteria. The original continuous-flow culture and a continuous-flow culture derived from a lyophilized sample of it that had been stored for 180 days at -70°C were provided to day-old chicks. Experiments utilizing the original culture were performed after 5, 30, 100, and 365 days of continuous-flow conditions and the lyophilized culture after 5 days of continuous-flow culture. In all experiments chicks were challenged with 10 4 CFU of Salmonella typhimurium at 3 days of age. At 10 days of age the chicks were killed and cecal contents analyzed for fermentation parameters and for S. typhumurium colonization. Ten-day-old chicks provided with the original or the lyophilized culture had significantly increased cecal propionic acid (P < 0.05) and decreased Salmonella cecal colonization (P < 0.05 ) compared to untreated controls in all experiments. The results indicate that continuous-flow cultures can be used as a practical method to maintain competetive-exclusion cultures without loss of efficacy, and continuous-flow cultures stored in a lyophilized form can be reconstituted and still maintain efficacy.

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.

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.

The objective of the current research was to determine if feeding the citrus by-products(D) -limonene (DL) and citrus molasses would reduce the concentration and prevalence of Salmonella in weanling pigs experimentally infected with Salmonella Typhimurium. Twenty crossbred weanling pigs (average body weight [BW], 19.9 kg) were randomly assigned to one of four treatments: control, low-dose DL (1.5 ml/kg of BW per day), high-dose DL (3.0 ml/kg of BW per day), and citrus molasses (0.05 kg/kg of BW per day). Treatments were administered in the feed (twice daily) for 7 days, with one-half of the dose administered at each feeding. Fecal samples were collected twice daily (prior to administration of treatment) and cultured for quantitative and qualitative determination of the challenge strain of Salmonella. Upon termination of the study, pigs were euthanized and tissues from the stomach, ileum, cecum, spiral colon, and rectum, as well as luminal contents, were collected. In addition, the popliteal and ileocecal lymph nodes and liver, spleen, and tonsil tissue were collected for qualitative Salmonella culture. No significant treatment differences (P0.05) were observed among treatments for fecal concentration or prevalence of Salmonella throughout the 7-day collection period. Likewise, no treatment differences (P0.05) were observed for any of the tissue or luminal content samples collected. Salmonella was not cultured from the muscle-bound popliteal lymph node but was cultured from the mesenteric ileocecal lymph nodes. While there were no effects in the current experiment, future research may examine the effect of a lower challenge dose and/or different administration (dose or duration) of the citrus by-products.

Our objective in this study was to use batch culture to estimate growth kinetic parameters for a growth-limiting amino acid, serine, in Salmonella typhimurium and a in chicken cecal bacterium, Escherichia fergusonii , and to test these predictions for competitiveness in a mixed culture of the two organisms. Under anaerobic growth conditions, the two bacteria grew only when serine was provided as the nitrogen source, When serine was used as a carbon source in aerobic media, the maximum growth rates of the two organisms were considerably lower and the affinity constants were higher than when serine was used as a nitrogen source. The maximum growth rates of the two organisms were lower in anaerobic media than in aerobic media, but serine anaerobic affinity constants were lower than those from aerobic media. In aerobic mixed culture, S. typhimurium outgrew E. fergusonii in N-limited minimal media containing 0.1 mM serine but in N-nonlimited minimal media containing 10 mM serine, E. fergusonii outgrew S. typhimurium . In anaerobic mixed culture, E. fergusonii outgrew S. typhimurium in media containing both 0.1 mM and 10 mM serine concentrations. It appears that the cause of inhibition of Salmonella with serine as the variable nutrient differs depending on the oxidation-reduction status.

Experiments were conducted to examine the effect of a continuous-flow (CF) derived bacterial culture and of dietary lactose on colonization level by anaerobic bacteria in the ceca of broiler chicks. Cecal facultative and strictly anaerobic bacteria colony forming units (CFU), total volatile fatty acid (TVFA), lactic and propionic acid concentrations in 3-d-old chicks, and cecal log10 Salmonella typhimurium colonization and propionic acid concentrations in 10-d-old chicks were measured. Treatment groups were control diet, 2% lactose diet, CF culture + control diet, and CF culture + 2% lactose diet. Groups inoculated with CF culture had decreased (P < 0.05) Salmonella in cecal contents at 10 d of age. The level of Salmonella protection was significantly (P < 0.05) correlated with increased cecal bacteria CFU, TVFA, and propionic acid concentrations in 3-d-old chicks. Results indicated that inoculating newly hatched chicks with CF culture containing cecal microflora originally obtained from adult chickens, facilitates early cecal colonization by native cecal microflora, and this is associated with protection against Salmonella .

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.

Information implicating bacterial biofilms as contributory factors in the development of environmental bacterial resistance has been increasing. There is a lack of information regarding the role of biofilms within the microbial ecology of the gastrointestinal tract of food animals. This work used a continuous-flow chemostat model derived from the ceca of 7-day-old chicks to characterize these communities and their ability to neutralize invasion by Salmonella enterica serovar Typhimurium. We characterized and compared the biofilm and planktonic communities within these microcosms using automated ribotyping and the Analytical Profile Index biotyping system. Eleven species from eight different genera were identified from six culture systems. Klebsiella pneumoniae was isolated from all planktonic communities and four of the biofilm communities. Three of the communities resisted colonization by Salmonella enterica serovar Typhimurium, two communities suppressed growth, and one community succumbed to colonization. In cultures that resisted colonization, no Salmonella could be isolated from the biofilm; in cultures that succumbed to colonization, Salmonella was consistently found within the biofilms. This study was one of a series that provided a molecular-based characterization of both the biofilm and planktonic communities from continuous-flow culture systems derived from the cecal microflora of chicks, ranging in age from day-of-hatch to 14 days old. The one common factor relating to successful colonization of the culture was the presence of Salmonella within the biofilm. The capacity to sequester the introduced Salmonella into the biofilm appears to be a contributing factor to the inability of these cultures to withstand colonization by the Salmonella.