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Effects of subtherapeutic administration of antimicrobial agents to beef cattle on the prevalence of antimicrobial resistance in Campylobacter jejuni and Campylobacter hyointestinalis.

The influence of antimicrobial agents on the development of antimicrobial resistance (AMR) in Campylobacter isolates recovered from 300 beef cattle maintained in an experimental feedlot was monitored over a 315-day period (11 sample times). Groups of calves were assigned to one of the following antimicrobial treatments: chlortetracycline and sulfamethazine (CS), chlortetracycline alone (Ct), virginiamycin, monensin, tylosin phosphate, and no antimicrobial agent (i.e., control treatment). In total, 3,283 fecal samples were processed for campylobacters over the course of the experiment. Of the 2,052 bacterial isolates recovered, 92% were Campylobacter (1,518 were Campylobacter hyointestinalis and 380 were C. jejuni). None of the antimicrobial treatments decreased the isolation frequency of C. jejuni relative to the control treatment. In contrast, C. hyointestinalis was isolated less frequently from animals treated with CS and to a lesser extent from animals treated with Ct. The majority (> or =94%) of C. jejuni isolates were sensitive to ampicillin, erythromycin, and ciprofloxacin, but more isolates with resistance to tetracycline were recovered from animals fed Ct. All of the 1,500 isolates of C. hyointestinalis examined were sensitive to ciprofloxacin. In contrast, 11%, 10%, and 1% of these isolates were resistant to tetracycline, erythromycin, and ampicillin, respectively. The number of animals from which C. hyointestinalis isolates with resistance to erythromycin and tetracycline were recovered differed among the antimicrobial treatments. Only Ct administration increased the carriage rates of erythromycin-resistant isolates of C. hyointestinalis, and the inclusion of CS in the diet increased the number of animals from which tetracycline-resistant isolates were recovered. The majority of C. hyointestinalis isolates with resistance to tetracycline were obtained from cohorts within a single pen, and most of these isolates were recovered from cattle during feeding of a forage-based diet as opposed to a grain-based diet. The findings of this study show that the subtherapeutic administration of tetracycline, alone and in combination with sulfamethazine, to feedlot cattle can select for the carriage of resistant strains of Campylobacter species. Considering the widespread use of in-feed antimicrobial agents and the high frequency of beef cattle that shed campylobacters, the development of AMR should be monitored as part of an on-going surveillance program.

Animals↗

Development of a reverse transcription-nested polymerase chain reaction assay for differential diagnosis of transmissible gastroenteritis virus and porcine respiratory coronavirus from feces and nasal swabs of infected pigs.

Transmissible gastroenteritis virus (TGEV), a coronavirus, replicates in intestinal enterocytes and causes diarrhea in young pigs. Porcine respiratory coronavirus (PRCV), a spike (S) gene natural deletion mutant of TGEV, has a respiratory tissue tropism and causes mild or subclinical respiratory infections. Conventional antigen-based diagnostic tests fail to differentiate TGEV and PRCV, and a blocking ELISA test to serologically differentiate TGEV/PRCV-infected pigs is conducted on convalescent serum retrospectively after disease outbreaks. A reverse transcription (RT)-nested polymerase chain reaction (PCR) with primers targeted to the S gene deletion region to differentiate TGEV/PRCV was developed. The specificity of the RT-nested PCR was confirmed with reference and recent field strains of TGEV/PRCV, and its sensitivity was analyzed by testing nasal and fecal samples collected from pigs at various days postinoculation (DPI) with TGEV or PRCV. Specific PCR products for TGEV/PRCV were detected only with the homologous reference or field coronaviruses and for 10-14 DPI of pigs with TGEV (feces) or PRCV (nasal samples). The RT-nested PCR assay was more sensitive than antigen-based assays on the basis of duration of virus detection in experimentally infected pigs and was directly applicable to nasal as well as fecal specimens from the field.

Animals↗

Enumeration of Campylobacter spp. in broiler feces and in corresponding processed carcasses.

Twenty north Georgia commercial flocks of broiler chickens sampled in 1995 and 11 flocks sampled in 2001 were tested for Campylobacter spp. Direct plating on Campy-Cefex agar was carried out to determine levels of Campylobacter colonization within each flock through the enumeration of the organism in 50 fresh fecal samples 1 day prior to slaughter. The next morning, these flocks were the first to be processed, and levels of the organism per carcass before the chilling operation (50 carcasses per flock) in 2001 and after the chilling operation (50 carcasses per flock) in both 1995 and 2001 were estimated. Levels of the organism on freshly processed broiler carcasses were estimated by the same methods in 1995 and 2001, and a significant reduction from an average of 10(4.11) CFU per carcass in 1995 to an average of 10(3.05) CFU per carcass in 2001 was observed. Levels of Campylobacter spp. found in production and in processing were not strongly correlative, indicating the existence of complex parameters involving production factors and variables associated with flock transport and the processing of the broilers. The reduction in Campylobacter levels on processed carcasses may have contributed to the reduction in the frequency of human disease observed by the Centers for Disease Control during the same period. These data characterize the distribution of Campylobacter in north Georgia poultry operations and should assist in the development of risk assessment models for Campylobacter spp. The results obtained in this study suggest that the implementation of antimicrobial interventions by the poultry industry has already reduced consumer exposure to the organism.

Animals↗

Prevalence and diversity of Campylobacter jejuni in pig herds on farms with and without cattle or poultry.

The prevalence and diversity of Campylobacter jejuni was investigated in pig herds on farms with and without cattle or poultry production. A bacteriological screening of pig cecal samples from 247 finisher herds was carried out at the slaughter-house. Subsequently, a follow-up study was conducted in 24 herds (either with or without prior C. jejuni isolation from pigs) in which fecal samples were collected from pigs and, if present, cattle and poultry. Samples were analyzed for presence of Campylobacter, and subsequent analysis included species identification, serotyping, and, for selected strains, pulsed-field gel electrophoresis typing. In the slaughterhouse screening, C. jejuni was isolated from pigs in 21 (8.5%) herds, but no significant difference in prevalence was found between herd types (pigs, pigs and cattle, pigs and poultry). At the slaughterhouse, C. jejuni and Campylobacter coli prevalence in pigs was 2.3 and 90.1%, respectively. In the follow-up study, herd prevalence of C. jejuni was 8.3%, whereas C. jejuni and C. coli were isolated from 0.8 and 92.0% of pigs, respectively. In mixed production herds, C. jejuni predominated in cattle (42.7%) and poultry (31.6%), whereas C. jejuni was only isolated from 1.3 to 2.5% of pigs in these herds. There were no significant differences in C. jejuni or C. coli prevalence in pigs, cattle, and poultry between herds with and without prior C. jejuni isolation at the slaughterhouse. Pulsed-field gel electrophoresis typing did not yield evidence of C. jejuni transmission between cattle or poultry and pigs in mixed production herds. In contrast, pulsed-field gel electrophoresis analysis showed indistinguishable serotypes of C. coli in pigs and cattle in two herds. Verification of C. jejuni-positive pig samples showed that individual pigs can excrete high levels of C. jejuni and that mixed infection with C. jejuni and C. coli was common in C. jejuni-positive pigs. The results of our study suggest that transmission of C. jejuni between pigs and cattle or poultry in mixed production herds occurs infrequently. Detection of indistinguishable C. coli isolates in two herds, however, might indicate the existence of low-level transmission between pigs and cattle in herds of mixed production.

Abattoirs↗

Usefulness of a combination of pulsed-field gel electrophoresis and enrichment culture in laboratory investigation of a foodborne outbreak due to Clostridium perfringens.

Clostridium perfringens is ubiquitous in nature and normally detectable in human stools. Therefore, it is difficult to perform specific microbiologic diagnosis in foodborne outbreaks, particularly when only a few cultures are detected from fecal specimens. Usually, it has been necessary to detect over 10(6) spores/g of fecal sample as a diagnostic criterion of diarrhea due to C. perfringens. A relatively large foodborne outbreak occurred in Osaka City, Japan in October 2001. Although C. perfringens was suspected as the causal agent, four to seven days had passed after the onset of symptoms before fecal specimens were brought into our laboratory. The positive rate obtained by direct plating was quite low (13/83). We attempted to detect the organisms using enrichment culture after 75 degrees C 20 min heat-treatment, and C. perfringens enterotoxin gene (cpe)-positive strains were isolated from 53 of 81 samples. Pulsed-field gel electrophoresis (PFGE) and serotyping showed that 36 (67.9%) of these 53 strains had indistinguishable PFGE patterns and the same serotype, TW69. Our experience indicates that the enrichment culture could be useful for laboratory confirmation of a C. perfringens foodborne outbreak if it is used with adequate molecular epidemiologic methods.

Clostridium Infections↗

Parasitological analyses of the male chimpanzees (Pan troglodytes schweinfurthii) at Ngogo, Kibale National Park, Uganda.

Numerous intestinal parasites identified in populations of wild nonhuman primates can be pathogenic to humans. Furthermore, nonhuman primates are susceptible to a variety of human pathogens. Because of increasing human encroachment into previously nonimpacted forests, and the potential for disease transmission between human and nonhuman primate populations, further detailed investigations of primate ecological parasitology are warranted. For meaningful comparisons to be made, it is important for methods to be standardized across study sites. One aspect of methodological standardization is providing reliable estimates of parasite prevalence and knowing how many samples are needed to adequately estimate an individual's parasite prevalence. In this study the parasitic fauna of 37 adult, adolescent, and juvenile male chimpanzees from the Ngogo group, Kibale National Park, Uganda, were assessed from 121 fecal samples collected over a 3-month period. Twelve taxa of intestinal species (five helminth and seven protozoan) were recovered from the samples. The four most prevalent species were Troglodytella abrassarti (97.3%), Oesophagostomum sp. (81.1%), Strongyloides sp. (83.8%), and Entamoeba chattoni (70.3%). No one species was found in all samples from any one animal, and Troglodytella abrassarti, the most common intestinal organism, was found in all of the serial samples of only 69.4% of the chimpanzees. The cumulative species richness for individuals significantly increased for every sequential sample (up to three to four samples) taken per animal during this study. The results indicate that to accurately diagnose total intestinal infection and evaluate group prevalence, three to four sequential samples from each individual must be collected on nonconsecutive days. This conclusion applies only to short study periods in which possible seasonal effects are not taken into consideration. Validation of these results at different study sites in different regions with different climatic patterns is needed.

Animals↗

Chromosome breaking activity of human feces and its enhancement by transition metals.

Chloroform-methanol extracts from human fecal samples were found to contain compounds which induce chromosome aberrations in Chinese hamster ovary cells. The induction of chromosome aberrations is stimulated by Cu2+ or Mn2+ and inhibited by Fe2+ or Fe3+. Addition of catalase to the fecal extract or the mixture of fecal extract and Mn2+ reduced the frequency of chromosome aberrations. These properties are indicative of hydroxyradical producing agents.

Animals↗

Composition of fecal fiber from human subjects.

This research addresses two problems of fecal fiber analysis: the nitrogen in fecal neutral detergent fiber (NDF); and the impact of sodium sulfite on NDF yield and composition. Fecal samples for analysis were collected from healthy adults during a study of the effect of a wheat bran supplement on bowel function. When sodium sulfite, originally a component of neutral detergent solution, was omitted, the yield of fecal NDF, the concentration of apparent lignin in the NDF and the amount of nitrogen recovered in both the NDF residue and the lignin increased. These data support a previous report by Van Soest that the sulfite attacks lignin. We present evidence that the effect of sodium sulfite also could be one of inhibiting the formation of Maillard products, which are detected as lignin, during the reflux step of the NDF method. Three different fractions of nitrogen were identified in fecal NDF: the fraction eliminated by sodium sulfite; a fraction removed by dilute acid; and the fraction that remained in Klason lignin. Apparent NDF digestibilities were calculated several ways by using correction factors to account for this nitrogen. Only the subtraction of Klason lignin from fecal NDF eliminated the one negative digestibility we observed.

Aged↗

Specificities of the fecal microbiota in inflammatory bowel disease.

BACKGROUND: Abnormalities have been described in the fecal microbiota of patients with IBD, but it is not known whether they are specific for inflammatory bowel disease (IBD) or to some extent common to other forms of colitis. The aim of this study was to compare the bacterial composition of the dominant fecal microbiota in patients with Crohn's disease (CD), ulcerative colitis (UC), infectious colitis (IC), and in healthy subjects (HS). METHODS: Fluorescent in situ hybridization adapted to flow cytometry was used to analyze the bacterial composition of fecal samples from 13 patients with active CD, 13 patients with active UC, 5 patients with IC, and 13 HS. We used 6 group-specific probes targeting 16S rRNA and spanning the main phylogenetic groups of the fecal microbiota. RESULTS: A significantly higher proportion of the total fecal bacteria were recognized by the 6 probes in HS (86.6%+/-12.7) and in IC (84.0%+/-11.7) than in patients with IBD (70.9%+/-15 in CD and 60.1%+/-25.7 in UC). The Clostridium coccoides group was reduced in UC (20.0%+/-13.3 versus 42.0%+/-12.0 in HS; P<.001), whereas the C leptum group was reduced in CD (13.1%+/-11.9 versus 25.2%+/-14.2 in HS; P=.002). The Bacteroides group was more abundant in IC (36.4%+/-22.9) than in the other 3 groups (13.8%+/-11.8 in CD, 11.7%+/-11.7 in UC, 12.1%+/-7.0 in HS; P<.001 for all 3 comparisons). CONCLUSIONS: In IBD the dominant fecal microbiota comprises unusual bacterial species. Moreover, CD and UC fecal microbiota harbor specific discrepancies and differ from that of IC and healthy subjects.

Adult↗

Emergence of rotavirus G4P8 strain among children suffering from watery diarrhea in Calcutta, India.

Rotavirus RNAs from the fecal samples were hybridized with cDNAs specific for G1 and G2 genotypes. 59 out of 138 samples (42.7%) did not hybridize with either probe. The cDNAs coding for VP7 and VP4 from one such sample, SC134, were synthesized by combined reverse transcriptase/polymerase chain reaction (PCR) using specific oligonucleotide primers and were used as probes to screen those local isolates which did not hybridize with either G1 or G2. 26/59 (44%) of fecal RNA hybridized with these cDNAs indicating a possible emergence of this strain in this part of India. The VP7 and VP4 genotype specificity of SC134 was found to be G4P8 by multiplex PCR. The VP7 gene of SC134 was cloned and characterized in detail by restriction enzyme digestion and DNA sequence analysis. Comparison of nucleotide and predicted amino acid sequence of the VP7 gene of SC134 with other serotypes revealed that the VP7 gene of SC134 was closely related to G4. However, amio acids within the VP7 sequence differed in seven positions with that of both the subtype 'A' and 'B' of the G4 serotype. To establish the relation of this emerging strain with the other reported G4 strains, a phylogenetic tree was constructed and SC134 was found to be more closely related to the ST3 strain isolated in England and included in the tetravalent vaccine formulation along with two Japanese strains, although all four were distinct and did not form any cluster as was evident by the horizontal distance separating them. The VP7 gene sequence of SC134 was submitted to EMBL and was assigned the accession number AJ278217.

Antigens, Viral↗

[Rapid detection and identification of enteropathogenic bacteria by a single strain counting-PCR assay].

The arbitrarily primed-PCR (AP-PCR) method is a technique to generate patterns of randomly amplified DNA fragments, and is often used for typing clinical isolates. Currently, we have been developing a modified AP-PCR method, the single strain counting-PCR (SSC-PCR), in order to discriminate among bacterial strains based on amplified patterns with multiple primers. Here, we describe the availabilities of the SSC-PCR method to identify and detect enteropathogenic bacteria. The analysis of the amplified patterns of more than 100 bacterial species shows that the patterns among strains in a species are similar to one another, and that the pattern differs among species. These results indicate that the SSC-PCR method can be a novel identification system for enteropathogenic bacteria. To examine the possibility that we can detect enteropathogenic bacteria in feces using the SSC-PCR, the model specimens were made by mixing the Vibrio parahaemolyticus DNA and the DNA directly extracted from feces. Although the patterns of fecal samples are complex with many bands, we can find a specific band of V. parahaemolyticus in the patterns of model specimens. Thus, the SSC-PCR method may be available for rapid detection of enteropathogenic bacteria in fecal specimens.

DNA, Bacterial↗

Biodistribution and dosimetry of indium-111-polyclonal IgG in normal subjects.

UNLABELLED: Indium-111-polyclonal IgG is a new imaging agent of infection and inflammation that has been developed as a possible replacement for radiolabeled leukocytes. We undertook a study to determine the safety, biodistribution and dosimetry of the agent in normal subjects. METHODS: Twelve normal male volunteers with an average age of 34 yr (range 21-55 yr) were studied. Each was injected with 1.22-1.47 mCi 111In-labeled polyclonal IgG; digital whole-body images, in addition to blood, urine and fecal samples, were obtained immediately after injection and at 6, 24, 48, 72, 96 and 120 hr. Whole-body counts, as well as individual organ data obtained by outlining regions of interest, were measured. Blood, urine and fecal counting were done in a well counter and compared to known standards; dosimetry calculations were performed with the MIRD technique. RESULTS: The mean whole-blood activity had a two-phase disappearance curve: the T1/2I was 11.4 hr (61.1%) and the T1/2II was 112.5 hr (38%). Twelve percent of the dose was excreted in the urine and 1.14% in the feces. Skeletal muscle had the highest percentage of uptake, followed by the bone marrow, liver and lungs; the spleen showed less than 1% uptake. Activity in the lungs varied with time, falling by 37% after 18 hr and by 68% after 72 hr. Dosimetry calculations indicated that the highest absorbed dose was to the liver (1.42 rad/mCi) followed by the testes (1.23 rad/mCi) and red marrow (0.976 rad/mCi). The total-body dose was 0.467 rad/mCi, with an effective dose equivalent of 790.84 mrem. CONCLUSION: The biodistribution of 111In IgG is similar to that of 99mTc-HMPAO-labeled leukocytes. Activity in the liver, kidneys and GI tract may make evaluation of infection in these regions difficult. The dosimetry data indicate that adequate doses can be administered for clinical imaging without exposing the patient to excessive radiation.

Adult↗

Colonic proteinases: increased activity in patients with ulcerative colitis.

In order to study the colonic intraluminal proteinase-antiproteinase imbalance under inflammatory conditions, we determined proteolytic activity (PA), alpha-1-antitrypsin and the activities of trypsin, chymotrypsin and neutrophil elastase in feces from patients with ulcerative colitis (UC) comparing the results with a control group. A fecal sample was obtained from each 25 patients with ulcerative colitis and 10 control subjects were studied. The severity of the disease was assessed by the Truelove index. Proteolytic activity was measured using azocasein as proteolytic substrate. The fecal concentration of alpha-1-antitrypsin was measured by radial immunodiffusion and the activities of the enzymes were measured using specific substrates. We found an increase in fecal PA, alpha-1-antitrypsin and neutrophil elastase in patients with UC and the correlation between the severity of the disease and the PA was statistically significant (r = 0.62, P < 0.05). We conclude that elevated colonic proteinase activity could contribute to the pathophysiology of ulcerative colitis.

Chymotrypsin↗

A randomized, controlled, open-label trial of a single day of mebendazole versus a single dose of tinidazole in the treatment of giardiasis in children.

BACKGROUND: Giardia duodenalis is the most commonly detected parasite in the intestinal tract of humans and 5-nitroimidazole compounds, quinacrine and furazolidone have been used against giardiasis. However, cases refractory to treatment with these drugs are becoming more common worldwide. OBJECTIVE: To compare the efficacy and safety of mebendazole versus tinidazole in the treatment of giardiasis. RESEARCH DESIGN AND METHODS: 122 children (aged 5 to 15 years) of both sexes with confirmed Giardia duodenalis cysts or trophozoites in their stool samples were randomly separated into two groups of 61 individuals. Each group received either mebendazole 200 mg three times for 1 day or tinidazole 50 mg/kg in a single dose. The evaluation of the efficacy was based on parasitological response. Parents or legal guardians of each child were asked to provide three fecal samples on days 3, 5, and 7 after treatment completion. A child was considered to be cured if no Giardia trophozoites or cysts were found in any of the three post-treatment fecal specimens evaluated by direct wet mounts and/or after Ritchie concentration techniques. RESULTS: The frequency of cure was higher for tinidazole (81.97%) than for mebendazole (63.93%); the difference was statistically significant (p < 0.05). Transient abdominal pain was more common in children treated with mebendazole (p < 0.05), whereas loss of appetite, bitter taste, headache, vomiting, and nausea were more common in the tinidazole-treated group (p < 0.05). CONCLUSIONS: Three doses of mebendazole, in a single day, are inferior to a single dose tinidazole in the treatment of giardiasis.

Adolescent↗

Universal Identification of Pathogenic Viruses by Liquid Chromatography Coupled with Tandem Mass Spectrometry Proteotyping.

Accurate and rapid identification of viruses is crucial for an effective medical diagnosis when dealing with infections. Conventional methods, including DNA amplification techniques or lateral-flow assays, are constrained to a specific set of targets to search for. In this study, we introduce a novel tandem mass spectrometry proteotyping-based method that offers a universal approach for the identification of pathogenic viruses and other components, eliminating the need for a priori knowledge of the sample composition. Our protocol relies on a time and cost-efficient peptide sample preparation, followed by an analysis with liquid chromatography coupled to high-resolution tandem mass spectrometry. As a proof of concept, we first assessed our method on publicly available shotgun proteomics datasets obtained from virus preparations and fecal samples of infected individuals. Successful virus identification was achieved with 53 public datasets, spanning 23 distinct viral species. Furthermore, we illustrated the method's capability to discriminate closely related viruses within the same sample, using alphaviruses as an example. The clinical applicability of our method was demonstrated by the accurate detection of the vaccinia virus in spiked saliva, a matrix of paramount clinical significance due to its non-invasive and easily obtainable nature. This innovative approach represents a significant advancement in pathogen detection and paves the way for enhanced diagnostic capabilities.

Tandem Mass Spectrometry↗

Improvement of immunomagnetic separation for Escherichia coli O157:H7 detection by the PickPen magnetic particle separation device.

Conventional immunomagnetic separation (IMS) procedures, which use an external magnetic source to capture magnetic particles against the side of a test tube, are labor-intensive and can have poor sensitivity for the target organism because of high background microflora that is not effectively washed away during the IMS process. This report compares the conventional IMS procedure to a new IMS procedure with an intrasolution magnetic particle transfer device, the PickPen. The IMS target for the majority of these studies is Escherichia coli O157:H7 in various types of samples, including cattle feces, hides, carcasses, and ground beef. Comparison of the two IMS methods showed a significant difference (P < 0.05) in the efficiency of detecting E. coli O157:H7 from cattle carcass surface, cattle hide, and cattle fecal samples. No significant improvement (P > 0.05) in E. coli O157:H7 detection was observed when the PickPen IMS procedure was used to isolate this pathogen from ground beef samples. Use of the PickPen IMS greatly increases the throughput of the IMS procedure and may be more compatible with various emerging technologies for pathogen detection. In addition, the efficacy of sequential IMS for multiple pathogens is reported herein.

Animals↗

Fecal leukocytes and epithelial cells in horses with diarrhea.

Fecal samples from 74 horses with diarrhea were examined microscopically for leukocytes and epithelial cells. The diarrhea was categorized as either acute (less than two weeks) or chronic, Salmonella positive or negative, and mild or severe (based on the fecal consistency). A large number of fecal leukocytes was strongly suggestive of salmonellosis; however, approximately 1/3 of the horses with Salmonella-negative diarrhea also shed fecal leukocytes. Fecal leukocytes were more common in horses with acute and/or severe diarrhea regardless of cause. Numbers of fecal colonic mucosal epithelial cells were increased in severe diarrhea, and their presence in horses with chronic diarrhea was correlated with salmonellosis. The study supports the concept that fecal leukocytes and epithelial cells occur in increased numbers in equine salmonellosis but also occur in other types of diarrhea.

Animals↗

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

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

Animal Feed↗