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PubMed2004

Chronic shedding of Campylobacter species in beef cattle.

AIMS: To determine the prevalence of chronic shedding of Campylobacter species by beef cattle, a longitudinal study of shedding patterns was conducted in a cohort of 60 beef steers over a 4-month period. METHODS AND RESULTS: Steers were maintained in a simulated feedlot setting but individually in pens to minimize transmission among animals. At each collection time, campylobacters in faeces were detected using conventional PCR. In addition, quantities of Campylobacter jejuni and C. lanienae in faeces were measured using real-time quantitative (RTQ) PCR. All of the steers tested shed Campylobacter species during the course of the study, and overall, 90% of the 299 samples tested were positive for Campylobacter DNA. The majority of the animals (86%) shed campylobacters at >/=4 sample times. The most prevalent taxon detected in bovine faeces was C. lanienae (56% of samples) followed by C. jejuni (13%), C. hyointestinalis (8%), and C. fetus (2%). No C. coli was detected, and 13% of the faecal samples contained two or more of the above species. Seven (12%) and 34 (57%) animals shed C. jejuni and C. lanienae at >/=3 sample times, respectively. For both C. lanienae and C. jejuni, a substantial number of cells were detected in faeces using RTQ-PCR; 27% of the samples positive for C. jejuni contained populations >10(4) cells g(-1) (maximum of 5 x 10(5) cells g(-1)), and 44% of samples positive for C. lanienae possessed populations >10(6) cells g(-1) (maximum of 4 x 10(8) cells g(-1)). A significant correlation was observed between shedding of C. lanienae and the severity of liver abscesses. In 27% of the samples, an amplicon was obtained for genus-specific but not for the species-specific primers. Sequencing of the partial 16S rRNA gene suggested the presence of at least two undescribed Campylobacter species but this has yet to be confirmed. CONCLUSIONS: A high percentage of feedlot cattle shed large quantities of Campylobacter species in their faeces over a protracted period of time (ca 112 days). SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study of longitudinal shedding patterns of campylobacters in beef cattle using PCR-detection methods. In addition, this is the first use of RTQ-PCR to directly quantify C. jejuni or C. lanienae in faeces. The results of the study show that a large number of cattle (>85%) chronically shed campylobacters in feedlots.

Animals↗

PubMed2005

The occurrence of Campylobacter subtypes in environmental reservoirs and potential transmission routes.

AIM: To identify potential reservoirs and transmission routes of human pathogenic Campylobacter spp. METHODS AND RESULTS: An enrichment PCR method for the detection and identification of Campylobacter jejuni and/or Campylobacter coli in faecal, food and river water samples was applied to 1450 samples of 12 matrix types obtained from a defined geographical area. PCR-positive samples were cultured to yield isolates for typing, and the data for 616 C. jejuni isolates obtained. Serotyping and SmaI macrorestriction profiling using pulsed field gel electrophoresis revealed a high level of diversity within the isolates from each matrix. Campylobacter jejuni and C. coli subtypes indistinguishable from those obtained from human cases were detected in most of the matrices examined. No Campylobacter isolates were isolated from possum faeces. CONCLUSIONS: Ten of the 12 matrices examined may be involved in the transmission of human campylobacteriosis as they contained Campylobacter subtypes also isolated from clinical cases. SIGNIFICANCE AND IMPACT OF THE STUDY: Results indicate that, for this rural population, a range of potential transmission routes that could lead to campylobacteriosis exist. Their relative importance needs to be assessed from an exposure assessment standpoint.

Animals↗

PubMed2004

Impact of chronic catheterization and automated blood sampling (Accusampler) on serum corticosterone and fecal immunoreactive corticosterone metabolites and immunoglobulin A in male rats.

Jugular catheters were inserted in nine male rats under general isofluorane anesthesia and the catheters were connected to a commercially available computerized blood sampling device (Accusampler). Blood samples (150 microl) were collected every 4 h during the first 24 h after surgery and every 12 h during the following 72 h until 94 h after surgery, when the animals were killed. All fecal pellets were collected at blood sampling. Serum corticosterone and fecal concentrations of immunoreactive corticosterone metabolites and immunoglobulin A (IgA) were quantified by ELISAs. In blood, high corticosterone concentrations (>200 ng/ml) were recorded in the first samples obtained after surgery, but the concentrations decreased steadily during the day and became cyclical, showing a diurnal variation with high levels during evenings and low levels in the mornings. The automatic blood sampling itself did not result in recordable increases in serum corticosterone concentrations. The time delay between the presence of elevated corticosterone levels in blood and in feces was approximately 12 h. Fecal immunoreactive corticosterone metabolite levels remained elevated during the 94 h study period after surgery. The fecal concentrations of IgA showed substantial between-animal variation and decreased non-significantly after the surgery. Like serum corticosterone, fecal IgA showed a diurnal variation in amounts excreted, in this case with high values in the morning and low values in the evening. The concentrations of fecal corticosterone and IgA were negatively correlated in samples obtained before surgery but no correlation existed after surgery. This indicates that fecal immunoreactive corticosterone metabolites, but not IgA, constitute a good marker of acute stress. For immunoreactive corticosterone metabolites as well as for IgA, the concentration in feces correlated well with total excretion, making single fecal samplings usable as a measure of total secretion.

Acute Disease↗

PubMed2004

Variation in the faecal shedding of Salmonella and E. coli O157:H7 in lactating dairy cattle and examination of Salmonella genotypes using pulsed-field gel electrophoresis.

AIMS: To examine the variability in faecal shedding of Salmonella and Escherichia coli O157:H7 in healthy lactating dairy cattle and to evaluate the genetic relatedness of Salmonella isolates. METHODS: Faecal samples were obtained from lactating Holstein dairy cattle on four commercial farms in the southwestern US. All farms were within an 8-km radius and were sampled in August 2001, January 2002 and August 2002 (60 cows per farm per sampling; n = 720 total samples). Samples were cultured for E. coli O157:H7 and Salmonella and a portion of the recovered Salmonella isolates were examined for genetic relatedness using pulsed-field gel electrophoresis (PFGE). RESULTS: Faecal shedding of E. coli O157:H7 and Salmonella varied considerably between farms and at the different sampling times. Large fluctuations in the percentage of positive animals were observed from summer to summer for both of these pathogens. Similarly, Salmonella serotype and serotype prevalence varied from farm to farm and within farm from one sampling time to another. Multiple Salmonella genotypes were detected for a number of serotypes and identical genotypes were found on different farms with one genotype of Salmonella Senftenberg identified on three of the four farms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the wide variability in pathogen shedding within and among dairy farms all located in a small geographical region and highlights the complexity of pathogen control at the farm level.

Animals↗

PubMed2005

Evaluation of microbiological quality of coastal waters in Greece.

To evaluate the microbiological water quality of bathing sites along the Achaia coastline (south western Greece), a survey was conducted to determine the concentration of faecal bacterial and phage indicators as well as the presence of human viruses. Seawater samples (234) were collected from nine bathing sites on the Achaia coastline and were analysed for the presence of: total coliforms, faecal coliforms, faecal streptococci, Escherichia coli, somatic coliphages, F-RNA bacteriophages, bacteriophages infecting Bacteroides fragilis, enteroviruses, adenoviruses and hepatitis A viruses. Most of the bacteriological analysis results were in accordance with the European Union standards. In all sites, bacteriophages were detected occasionally. Enteroviruses and adenoviruses were detected in 24 samples (10.26%) and 37 samples (15.81%) respectively. No samples were positive for the presence of hepatitis A virus. The overall data indicates that bathing sites are impacted by human faecal material. Both bacterial indicators and phages have low predictive capability for the presence of human viruses in coastal waters. None of the environmental parameters analysed was strongly related to the presence of the indicator organisms and viruses. Appropriate and effective administrative measures that should be taken into account may be considered in order to improve water quality and reduce public health risk.

Bathing Beaches↗

PubMed2001

Clostridium difficile-associated diarrhea in a VA medical center: clustering of cases, association with antibiotic usage, and impact on HIV-infected patients.

A case-control study of patients with stools assayed for Clostridium difficile toxin over a 24-month period at a Veterans Affairs hospital found that the majority of cases (70.6%) occurred in temporal clusters. Clustering was particularly evident on a designated human immunodeficiency virus (HIV) unit. Thirty-four (75.5%) of 45 HIV-infected patients with C difficile-associated diarrhea (CDAD) died during their hospitalization. Third-generation cephalosporins were the antibiotics most strongly associated with CDAD.

Adult↗

PubMed2006

Modelling the fate of faecal indicators in a coastal basin.

The paper describes a modelling study of near-shore coastal waters, undertaken to assess the impact of various bacterial input loads on the receiving waters in a coastal basin in the UK. Total and faecal coliforms, used as the indicators for bathing water quality under the European Union (EU) Bathing Water Directive, were numerically modelled using a 2D depth integrated hydro-environmental model. Details are given of the governing equations and solution methods used in the numerical model, together with a discussion of the recent development in faecal bacterial indicator modelling. Details are also given of a field data collection exercise, which involved initially collecting existing information on effluent input loads and followed by an intensive field survey. Using the water quality model, the mortality rate of the pathogen bacteria was investigated. Three methods were used to represent the relationship between the decay rate and the level of solar radiation including: a constant decay rate, day- and night-time decay rates and a solar radiation related time varying decay rate. Relatively close agreement between model predicted and measured total and faecal coliform concentration distributions were obtained for different day- and night-time decay rates and time varying decay rates. No significant differences were found in the optimum decay rates for total and faecal coliform levels. Finally, the impact of the individual inputs on the bathing water quality of the basin was also statistically and numerically investigated. Results showed that the River Irvine was the most significant input during high river flows, and that under these conditions the bathing waters were likely to fail to comply with the European Union Bathing Water Directive. For base river flow conditions the Meadowhead effluent input was found to be critical for both total and faecal coliform level predictions.

Enterobacteriaceae↗

PubMed1988

Efficacy of early season anthelmintic treatment against gastrointestinal nematodes.

The efficacy of levamisole and ivermectin in multiple-dose regimes for the control of parasitic gastroenteritis in first-season grazing calves was evaluated on a dairy cattle farm in Belgium. Thirty-nine female Holstein crossbred calves were randomly divided into three groups. Paddock 1 was used for the controls, paddock 2 for the levamisole group (dosed at 3, 6 and 9 weeks after the start of grazing) and paddock 3 for the ivermectin group (dosed at 3 and 8 weeks after turn-out). The treatments were evaluated on the basis of live weight, faecal egg output, and serum pepsinogen levels. The impact of the therapeutic dosing at timed intervals during the first months of the grazing season was remarkable; egg output in the levamisole and ivermectin groups between June and early October was substantially lower. The treatments seem to adequately control Ostertagia, because serum pepsinogen values were much lower from August onwards. Better weight gains were observed in both the treatment groups. The experiment also illustrated the advantage of early housing of calves.

Animals↗

PubMed2026

Copper is an intestinal habitat filter affecting the gut microbiota interactions with Salmonella Typhimurium.

BACKGROUND: Foodborne pathogens, including Salmonella enterica serovar Typhimurium (S. Typhimurium), pose a significant threat to both human health and livestock productivity. The pandemic S. Typhimurium ST34 clone acquired a genomic island (SGI-4) conferring high copper resistance, an adaptation relevant in the context of the widespread use of copper sulphate at therapeutic levels in pig farming. We investigated how high dietary copper influences the piglet gut microbiota and Salmonella-microbiota interactions that may explain the global spread of S. Typhimurium ST34. RESULTS: An on-farm study combined with faecal shotgun metagenomics revealed that several potential Salmonella competitor species, including Bifidobacterium, Escherichia, and Lactobacillus, were less abundant in piglets on high-copper diets. Anaerobic and aerobic culturing alongside whole genome sequencing of 131 species and copper sulphate susceptibility testing identified copper resistance gene acquisition in selected microbes, particularly within Escherichia. Niche competition assays demonstrated that copper resistance is critical for inter-species competition under high-copper conditions, with Salmonella's Type VI Secretion System providing a distinct advantage over Escherichia in the copper-modified niche. CONCLUSIONS: Our findings suggest that copper supplementation alters the piglet gut environment, impacting competitive dynamics between pathogenic and commensal bacteria, likely to influence the zoonotic transmission of pathogens. Video Abstract.

Animals↗

PubMed2006

Determining diet composition on complex swards using n-alkanes and long-chain fatty alcohols.

We conducted an experiment to quantify the accuracy of methods based on n-alkanes and long-chain fatty alcohols for determining the diet composition of animals grazing complex swards. We cut forage from two indigenous vegetation communities, a Molinia caerulea-dominated grassland and a Calluna vulgaris-dominated dwarf-shrub community, and offered it to mature ewes in different ratios in a zero-grazing experiment. Nine dietary categories were identified within the forage offered: Molinia caerulea, Festuca spp., Juncus effusus, Carex spp., Calluna vulgaris, Erica tetralix, Vaccinium myrtillus, and dead grass. Samples of each of these categories together with fecal samples from each individual animal were analyzed for n-alkane and long-chain fatty alcohol concentrations. We analyzed the data using optimization software to minimize the sum of squares differences in the proportional profiles of n-alkanes and fatty alcohols in the diet and feces. Different combinations of n-alkane and fatty alcohols were investigated to assess which gave the most accurate measures of diet composition from the fecal profile. The most accurate estimates were obtained using combinations of the n-alkanes C25, C29, C31, and C33 and the long-chain fatty alcohols 1-C24-ol, 1-C28-ol, and 1-C30-ol, and these gave values for Lin's concordance correlation coefficient between estimated and actual values of >0.98. Our results demonstrate that n-alkanes and long-chain fatty alcohols can be used to estimate several components within the diet of animals grazing complex swards. Diet composition information obtained using this methodology has wide-ranging applications in terms of the assessment of the impact of grazing animals on particular ecosystems or the quantification of nutrient supply to the animal from different selection choices.

Alkanes↗

PubMed2000

Absence of obvious short-term impact of the nematode-trapping fungus Duddingtonia flagrans on survival and growth of the earthworm Aporrectodea longa.

The nematode-trapping fungus Duddingtonia flagrans may be used in biological control of parasitic nematode larvae in faeces of domestic host animals after feeding the hosts with fungal chlamydospores. In this experiment a possible undesirable fungal impact on earthworms, of the species Aporrectodea longa, was investigated. As earthworms eat animal faeces, D. flagrans may come into contact with earthworms both in their alimentary tract and on their body surface. However during the experimental period of 20 days, when earthworms were living in soil and eating cattle faeces that were heavily infested with viable chlamydospores of D. flagrans there were no indications of internal or external mycosis among the earthworms.

Animals↗

PubMed2006

Analysis of fecal microbial flora for antibiotic resistance in ceftiofur-treated calves.

To evaluate the impact of ceftiofur treatment in calves on fecal shedding of ceftriaxone-resistant bacteria, 3 female Holstein dairy calves were treated by intramuscular injection with EXCENEL RTU (ceftiofur hydrochloride, Pharmacia and Upjohn) at a therapeutic dosage of 2.2 mg/kg/day for 5 consecutive days following label directions. Three untreated calves were housed separately and served as controls. One to 3 days following the initial administration of ceftiofur, there was a 14% and 2% increase of fecal bacteria resistant to 16 and 64 microg ceftriaxone/mL, respectively. This response remained unchanged from days 6 to 13, and increased resistance was seen at day 17. Randomly selected isolates of gram-positive and gram-negative bacteria with elevated resistance to ceftriaxone (minimal inhibitory concentration (MIC) >or=64 microg ceftriaxone/mL) were isolated from calf feces and identified. In vitro conjugation experiments revealed that both the ceftriaxone-resistance gene bla (CMY-2) and class 1 integron were transferred from two bacterial species to Salmonella spp. at a frequency of 10(7) to 10(5). MIC data revealed that Salmonella transconjugants acquired either reduced susceptibility or resistance to ceftriaxone as well as to multiple antibiotics. This genetic transfer occurred both within and between genera. Treatment of calves with therapeutic dosages of ceftiofur can significantly increase for at least 17 days following the initial treatment the fecal excretion of ceftriaxone-resistant bacteria, including Salmonella species.

Animals↗

PubMedDate not supplied

Felbamate metabolism in the rat, rabbit, and dog.

Two major and one minor metabolite of felbamate (FBM) as well as unchanged drug were isolated and identified by electron impact and chemical ionization mass spectrometry from rat and dog urine after dosing with [14C]FBM. The metabolites were 2-(4-hydroxyphenyl)-1,3-propanediol dicarbamate (p-OHF), 2-hydroxy-2-phenyl-1,3-propanediol dicarbamate, and 2-phenyl-1,3-propanediol monocarbamate. The metabolites and FBM were excreted mainly in urine, where their sum accounted to 81-94% of the radioactivity in hydrolyzed rat urine samples, 71-82% in rabbit urine samples, and 69-83% in dog urine samples. The amount of metabolites in the conjugated form was estimated to be 20-35% in rat, 20-30% in rabbit, and 10-20% in dog urine. The major biliary metabolite in all three species was p-OHF, whereas the amount of FBM was small. Metabolites found in dog feces were the same as those in the urine.

Animals↗

PubMed2006

Potential microbiological contamination of effluents in poultry and swine abattoirs.

Health risks in the effluents of seven swine abattoirs and of seven poultry abattoirs were evaluated with regard to environment degradation and to dissemination of pathogenic microorganisms during the rainy and dry seasons. Supply-water samples from affluents and effluents of the treatment systems at different sites within the abattoir processing system were analysed. Similarly, water samples from the three recipient sites (emission point, 100 m upstream, 100 m downstream) were also analysed. Temperature, free residual chlorine (FRC), total coliform bacteria, Escherichia coli, enterococci, identification and serotyping of salmonellae were assessed. Scalding is the most significant stage in the slaughtering chain (P<0.05) when temperature is taken into account. Temperatures at effluents and at the sampled sites in the water bodies accorded to state and federal legislation standards. Supply waters did not meet the standards for FRC and microbial count standards according to the Ministry of Health and within limits imposed by the Industrial and Sanitary Inspection Regulations for Animal Products. Feather plucking and evisceration in poultry slaughter and the cleansing of carcasses and facilities in poultry and swine slaughtering had the highest contamination impact. The three loci at the water bodies were above the microbiological standards for classes II and III sites, in conformity with Law 8468 of the state of São Paulo, Brazil and Conama. Salmonella was found at several sites during slaughter, at both types of abattoirs, including in the effluent treatment system. This showed that these sites were the dissemination sources of the microorganism.

Abattoirs↗

PubMed2003

A comparison of two pre-enrichment media prior to immunomagnetic separation for the isolation of E. coli O157 from bovine faeces.

AIMS: To compare the sensitivity of two pre-enrichment broth media prior to immunomagnetic separation for the isolation of Escherichia coli O157 from cattle faeces. METHODS AND RESULTS: One-gram portions of 721 cattle faeces collected from 43 farms were pre-enriched in buffered peptone water containing vancomycin, cefixime and cefsulodin (BPW-VCC) and buffered peptone water without additives (BPW-WOA), respectively. A total of 137 samples were positive for E. coli O157: 127 pre-enriched with BPW-WOA and 89 pre-enriched in BPW-VCC. Representative isolates were tested for phage type, verotoxin and eae (E. coli attaching and effacing) gene sequences, resulting in the recognition of eight different types. All the E. coli O157 types recognized were isolated by both methods except for three different strains, each of which were isolated only on a single occasion: two by BPW-WOA and another by BPW-VCC. CONCLUSIONS: The results clearly demonstrate, under the conditions of this study, that BPW without antibiotics was the superior pre-enrichment medium for the isolation of E. coli O157 from cattle faeces. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of BPW-WOA in preference to BPW-VCC for the isolation of E. coli O157 from cattle faeces in future research and outbreak studies should lead to a higher number of positive isolates.

Animals↗

PubMed2003

Detection of Escherichia coli serogroups O26, O103, O111 and O145 from bovine faeces using immunomagnetic separation and PCR/DNA probe techniques.

AIMS: The aim of this study was to isolate Escherichia coli O26, O103, O111 and O145 from 745 samples of bovine faeces using (i) immunomagnetic separation (IMS) beads coated with antibodies to lipopolysaccharide, and slide agglutination (SA) tests and (ii) PCR and DNA probes for the detection of the Verocytotoxin (VT) genes. METHODS AND RESULTS: IMS-SA tests detected 132 isolates of presumptive E. coli O26, 112 (85%) were confirmed as serogroup O26 and 102 had the VT genes. One hundred and twenty-two strains of presumptive E. coli O103 were isolated by IMS-SA, 45 (37%) were confirmed as serogroup O103 but only one of these strains was identified as Verocytotoxin-producing E. coli (VTEC). Using the PCR/DNA probe method, 40 strains of VTEC O26 and three strains of VTEC O103 were isolated. IMS-SA identified 21 strains of presumptive E. coli O145, of which only four (19%) were confirmed as serogroup O145. VTEC of this serogroup was not detected by either IMS-SA or PCR/DNA probes. E. coli O111 was not isolated by either method. CONCLUSION: IMS beads were 2.5 times more sensitive than PCR/DNA probe methods for the detection of VTEC O26 in bovine faeces. SIGNIFICANCE AND IMPACT OF THE STUDY: IMS-SA is a sensitive method for detecting specific E. coli serogroups. However, the specificity of this method would be enhanced by the introduction of selective media and the use of tube agglutination tests for confirmation of the preliminary SA results.

Agglutination Tests↗

PubMed2003

Antimicrobial susceptibility and factors affecting the shedding of E. coli O157:H7 and Salmonella in dairy cattle.

AIMS: To examine factors affecting faecal shedding of the foodborne pathogens Escherichia coli O157:H7 and Salmonella in dairy cattle and evaluate antimicrobial susceptibility of these isolates. METHODS: Faecal samples were obtained in replicate from lactating (LAC; n = 60) and non-lactating (NLAC; n = 60) Holstein cattle to determine influence of heat stress, parity, lactation status (LAC vs NLAC) and stage of lactation [</=60 or >60 days in milk (DIM)] and cultured for E. coli O157:H7 and Salmonella. A portion of the recovered isolates were examined for antimicrobial susceptibility using the broth microdilution technique. RESULTS: No effects of heat stress were observed. Lactating cows shed more (P < 0.01) E. coli O157:H7 than NLAC cows (43% vs 32%, respectively). Multiparous LAC cows tended to shed more (P = 0.06) Salmonella than primiparous LAC cows (39% vs 27%, respectively). Parity did not influence (P > 0.10) bacterial shedding in NLAC cows. Cows </=60 DIM shed more (P < 0.05) Salmonella than cows >60 DIM. Seventeen Salmonella serotypes were identified with the most prevalent being Senftenberg (18%), Newport (17%) and Anatum (15%). Seventy-nine of the Salmonella isolates were resistant to at least one of the seven antibiotics. Escherichia coli O157:H7 isolates were resistant to 11 different antibiotics with multiple resistance to nine or more antibiotics observed in five isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated differences in the shedding patterns of foodborne pathogens due to the stage of the milk production cycle and may help identify times when on-farm pathogen control would be the most effective.

Animals↗

PubMed1985

The impact of routine isolation techniques for Campylobacter jejuni in stool cultures.

Campylobacter jejuni is recognized as a leading cause of bacterial diarrhea. We studied the impact of performing routine isolation techniques for C jejuni in stool cultures in a community hospital. Since instituting such techniques, we have found C jejuni to be the leading cause of bacterial diarrhea at our hospital. Clinical and laboratory manifestations and treatment did not distinguish patients with C jejuni from patients with Salmonella species isolated from the stool. We believe that microbiology laboratories should incorporate routine measures for detecting C jejuni in fecal specimens.

Bacteriological Techniques↗