Search

A series of experiments were conducted to evaluate the effects of ergot alkaloids (dihydroergotamine, ergonovine, and ergotamine) on E. coli O157:H7 in both pure and mixed ruminal fluid culture. Alkaloids were added to solutions of E. coli O157:H7 strains 933 (pure and ruminal cultures) and 6058 (ruminal culture only), and growth rates and colony-forming units (CFU) of E. coli O157:H7 were measured. Two mixtures of all three alkaloids at either 2 or 500 microM for each alkaloid decreased (p < 0.001) the growth rate of E. coli O157:H7 in pure culture compared to the individual alkaloids. Dihydroergotamine tended (p = 0.07) to reduce growth rate of E. coli O157:H7 in pure culture compared with ergonovine or ergotamine alone. Increased concentrations of dihydroergotamine and ergotamine decreased (p < 0.003) growth rate of E. coli O157:H7 but increasing concentrations of ergonovine did not influence (p > 0.10) E. coli O157:H7 growth rate. Similar to results in pure culture, a mixture of all three alkaloids at various concentrations for each alkaloid decreased (p < 0.001) the CFU of E. coli O157:H7 strain 6058 in mixed ruminal culture compared to the individual ergot alkaloids. Dihydroergotamine decreased (p = 0.04) CFU of E. coli O157:H7 strain 6058 when compared to ergonovine but CFU were similar (p > 0.10) between dihydroergotamine and ergotamine. Ruminal and (or) intestinal populations of E. coli O157:H7 may be influenced in livestock consuming endophyte-infected tall fescue, and these alterations could be due to the presence of ergot alkaloids in fescue plants.

Orange peel and orange pulp are by-products that are included in feedlot and dairy cattle diets because of their low cost and high nutritional quality. The antimicrobial activity of citrus oils has been reported previously. The present study was carried out to determine whether these citrus by-products exert antimicrobial effects on Escherichia coli O157:H7 and Salmonella Typhimurium populations that are found in cattle gastrointestinal tracts. The growth of pure cultures (n = 3) of E. coli O157:H7 and Salmonella Typhimurium were reduced (p < 0.05) by addition of 2% (w/v) orange pulp and orange peel. Ruminal fluid was collected from cattle (n = 2) and E. coli O157:H7 or Salmonella Typhimurium were added. The addition of orange pulp and peel to in vitro mixed ruminal microorganism fermentations (n = 3) demonstrated that both orange pulp and peel reduced E. coli O157:H7 and Salmonella Typhimurium populations at least 2 log(10) in mixed ruminal fluid fermentations. Addition of orange pulp reduced (p < 0.05) E. coli O157:H7 populations from 10(5) to 10(2) colony-forming units (CFU)/mL and Salmonella Typhimurium populations (p < 0.05) from 10(4) to 10(2) CFU/mL. These results indicate that orange pulp and/or peel included in ruminant diets could decrease ruminal populations of foodborne pathogenic bacteria. Further research is needed to determine whether the antimicrobial activity of orange products against E. coli O157:H7 or Salmonella Typhimurium is expressed in the lower gastrointestinal tract.

Escherichia coli O157:H7, Salmonella, and Listeria are foodborne pathogens of critical importance that often colonize cattle. E. coli O157:H7 can be specifically killed by lytic bacteriophage, and lytic bacteriophage treatment has been suggested as a pre-harvest intervention strategy to reduce foodborne pathogens in cattle. To date, no systematic approach to determine the incidence of E. coli O157:H7-infecting lytic bacteriophage has been published. Therefore, the current study was designed to determine (1) the incidence of E. coli O157, Salmonella spp., and Listeria and (2) the incidence of E. coli O157:H7-infecting bacteriophage in the feces of feedlot steers in commercial feedlots in the United States. Fecal samples (n=60) were collected from four feedlots in two Southern Great Plains states (total (n=240 fecal samples). Salmonella and E. coli O157:H7 were found in 3.8% and 11.7% of the fecal samples, respectively. Bacteriophage targeting E. coli O157:H7 were found in all four feedlots, in 15% of the individual fecal samples, and in 55% of the cattle pens. Our results indicate that such bacteriophage are widespread in feedlot cattle, suggesting that further research into the ecological role of bacteriophage in the gastrointestinal tract is needed.

To examine the effect of ionophore supplementation on fecal shedding of Escherichia coli O157:H7 and Salmonella, crossbred beef calves (n=113; mean body weight [BW], 243 kg) were fed a mineral supplement with ionophore (1.76 g lasalocid/kg) for 61 days (d). Control calves received an identical mineral supplement without lasalocid. Calves were pastured on fescue/bermudagrass paddocks and supplemented with a corn/wheat midds/soybean meal supplement (1.5% of BW/d). Upon arrival, cattle were fed a commercial receiving ration containing 1 g chlorotetracycline/kg for 10 d. Sick calves were administered one or a combination of the following: Nuflor (florfenicol), Baytril (eurofloxacin), Micotil (tilmicosin), or LA 200 (oxytetracycline). Fecal samples were collected immediately prior to ionophore supplementation, approximately midway and at the end of the experimental period (60 d total ionophore feeding) for isolation of E. coli O157:H7 and Salmonella. Putative fecal coliforms were also isolated at these sampling times and examined for antimicrobial susceptibility. The study was replicated over a two year period (year 1, n=53 head; year 2, n=60 head). Ionophore supplementation had no effect (p>0.10) on the incidence of calves shedding E. coli O157:H7 or Salmonella. The percentage of calves shedding E. coli O157:H7 varied throughout the experimental period from 0 to 30%, while Salmonella was cultured from only three calves over the 2-year experimental period. Antimicrobial susceptibility profiles of putative fecal coliforms were consistent with antibiotic treatments administered during the study (observed resistance to chlortetracycline, florfenicol, oxytetracycline), while only one treatment effect was observed. Ionophore treatment resulted in a significantly higher number of coliform isolates resistant to ampicillin compared to controls in year 1, but not year 2. A number of fecal coliform isolates demonstrated resistance to multiple antibiotics, however, this was not affected (p>0.10) by ionophore supplementation. Mineral intakes, BW gain, and the number of sick calves were similar (p>0.10) among treatments. Ionophore supplementation had no affect on fecal shedding of E. coli O157:H7 or Salmonella and a negligible impact on antimicrobial susceptibility patterns of fecal coliforms in beef calves.

Temperate bacteriophages have always been central to the evolution of bacteria, although their importance has been consistently underestimated compared to transformation and conjugation. In the last 20 years, as more gene and genome sequences have become available and researchers have more accurately determined bacteriophage populations in the environment, we are gaining a clearer picture of their role in the past and potential role in the future. The transductive and lysogenic capacities of this class of bacteriophages have contributed to the evolution and shaping of emerging foodborne pathogenic bacteria through the dissemination of virulence and antibiotic resistance genes. For example, the genome sequences of Shigella dysenteriae, Escherichia coli O157:H7, and the Stxencoding bacteriophages demonstrate the critical role bacteriophage-mediated gene transfer events played in the evolution of these high-profile human pathogens. In this review, we describe the basic genetic exchange mechanisms mediated by temperate bacteriophages and how these mechanisms have been central to the dissemination of virulence genes, such as toxins and antibiotics from one species to another (the shiga-like toxins, and multiple antibiotic resistance dissemination in Salmonella are used as specific examples). Data demonstrating the role of bacteriophages in the spread of antimicrobial resistance in bacteria, including interspecies transduction, are also presented. That temperate bacteriophages play a role in the on-going evolution of emerging pathogenic bacteria is obvious, but it is also clearly an on-going process with a breadth that must be appreciated as well as studied further if we are to be able to foresee what new challenges will arise to imperil food safety.

Swine can harbor Salmonella in their gastrointestinal tracts. It has been estimated that up to 48% of the U.S. swine herd may carry Salmonella. Housing sows in farrowing stalls has become controversial due to animal welfare-based criticisms. An alternative production system is to keep sows outdoors on pasture with access to individual farrowing huts. This study was designed to determine the effects of two production systems on indicator bacteria and Salmonella of sows housed indoors in farrowing stalls (n = 52) compared to sows housed outdoors (n = 52) in English style huts. Each farrowing radial contained one wallow, from which mud (n = 290) and water (n = 290) samples were collected weekly. All samples were analyzed for generic E. coli, coliforms and Salmonella. No differences (p > 0.05) were detected in Salmonella, generic E. coli and coliform populations between indoor farrowing stalls and outdoor farrowing huts. However, all 8 outdoor wallows contained Salmonella spp. at some point during the study (n = 49 Salmonella isolates). Salmonella genotypes persisted within some wallows for >5 months, and genetically indistinguishable Salmonella isolates were found in multiple wallows. Salmonella isolated from outdoor sow feces were genetically indistinguishable by PFGE from Salmonella isolated from wallows (n = 33) throughout the study, indicating that pathogenic bacteria were cycling between swine and their environment. In conclusion, the role of wallows in disseminating Salmonella within an outdoor swine herd appears to be significant.

Mature, healthy lactating dairy cattle were sampled on two farms in the southwestern United States to examine the effects of heat stress (Experiment I) and stage of lactation (Experiment II) on the fecal shedding of E. coli O157:H7 and Salmonella. To examine the effects of heat stress, fecal samples were collected from 45 cows at 7:00 AM (coolest part of the day) and 5:00 PM (hottest part of the day) in August 2002 on a 250 cow dairy. The study was replicated one month later (n = 170 total samples). A temperature-heat index (THI) was calculated for each sampling time. In Experiment II, stage of lactation was examined by sampling lactating dairy cattle early [< 60 days in milk (DIM)] and late (> 150 DIM) in the lactation cycle in the summer of 2001. The study was replicated the following summer (60 cows/group/replicate; n = 240 total samples). For Experiment I, THI averaged 75 and 82 for the AM and PM samplings, respectively, indicating the cows were beginning to experience heat stress in the morning and by afternoon were in severe heat stress. The shedding of E. coli O157:H7 tended to be higher (p = 0.09) in the afternoon sampling of the first replicate, however was not different in the second replicate or when both replicates were pooled (p > 0.10). Salmonella shedding was not different (p > 0.10) at any sampling time with nearly 100% of the cows positive. Stage of lactation had no effect on the number of cows shedding E. coli O157:H7 (p > 0.10). Salmonella shedding tended to be higher (p = 0.09) in early lactation cows in the first replicate, while in the second replicate more late lactation cows were shedding Salmonella (p < 0.05); however, there were no differences due to stage of lactation when replicates were pooled (p > 0.10). While further research is needed, results of this research highlight the variability in pathogen shedding in healthy dairy cattle and indicate that environmental factors and/or production demands may influence shedding patterns of E. coli O157:H7 and Salmonella.

The spread of antibiotic resistance from farm animals to human pathogens has become a matter of great public health significance, and methods to reduce foodborne pathogenic bacteria without utilizing traditional antibiotics have been sought. Chlorate kills Escherichia coli O157:H7 in vitro and in vivo and has been proposed as a feed additive to be included in food animal rations immediately prior to slaughter to reduce E. coli O157:H7 populations in the animal gastrointestinal tract. This study was designed to examine the effect that the development of chlorate resistance in E. coli O157:H7 has on resistance to 32 medically important antibiotics. Chlorate resistance numerically increased antibiotic resistance in E. coli O157:H7 strain 6058 and strain 933 to six and four antibiotics, respectively, but chlorate resistance affected resistance to only two antibiotics (cefoxotin and sulfadimethoxine) in both strains examined. Based on the fact that chlorate resistant mutants do not appear to compete well in vivo and based on the results of the current study, it appears unlikely that chlorate treatment will result in a dissemination of antibiotic resistance. However, further research is needed to confirm these results.