PubMed Health⌕ Search

Biomedical subjects

G Daube

Publications and source records attributed to G Daube.

At least 19 recordsLinked to original sources
PubMed2006

Quantitative risk assessment of Campylobacter spp. in poultry based meat preparations as one of the factors to support the development of risk-based microbiological criteria in Belgium.

The objective of this study was to do an exercise in risk assessment on Campylobacter spp. for poultry based meat preparations in Belgium. This risk assessment was undertaken on the demand of the competent national authorities as one of the supportive factors to define risk-based microbiological criteria. The quantitative risk assessment model follows a retail to table approach and is divided in different modules. The contamination of raw chicken meat products (CMPs) was represented by a normal distribution of the natural logarithm of the concentration of Campylobacter spp. (ln[Camp]) in raw CMPs based on data from surveillance programs in Belgium. To analyse the relative impact of reducing the risk of campylobacteriosis associated with a decrease in the Campylobacter contamination level in these types of food products, the model was run for different means and standard deviations of the normal distribution of the ln[Camp] in raw CMPs. The limitation in data for the local situation in Belgium and on this particular product and more precisely the semi-quantitative nature of concentration of Campylobacter spp. due to presence/absence testing, was identified as an important information gap. Also the knowledge on the dose-response relationship of Campylobacter spp. was limited, and therefore three different approaches of dose-response modelling were compared. Two approaches (1 and 2), derived from the same study, showed that the reduction of the mean of the distribution representing the ln[Camp] in raw CMPs is the best approach to reduce the risk of Campylobacter spp. in CMPs. However, for the simulated exposure and approach 3 it was observed that the reduction of the standard deviation is the most appropriate technique to lower the risk of campylobacteriosis. Since the dose-response models used in approach 1 and 2 are based on limited data and the reduction of the mean corresponds with a complete shift of the contamination level of raw CMPs, demanding high efforts from the poultry industry, it is proposed to lower the standard deviation of the concentration of Campylobacter spp. in raw CMPs. This proposal corresponds with the elimination of the products that are highly contaminated. Simulation showed that eating raw chicken meat products can give rise to exposures that are 10(10) times higher than when the product is heated, indicating that campaigns are important to inform consumers about the necessity of an appropriate heat treatment of these type of food products.

Animals↗
PubMed2006

Prevalence of enterohaemorrhagic Escherichia coli from serotype O157 and other attaching and effacing Escherichia coli on bovine carcasses in Algeria.

AIMS: Bovine meat is the principal source of human contamination of attaching and effacing Escherichia coli, including enterohaemorrhagic E. coli O157. The aim was to study the prevalence of these strains on bovine carcasses in Algeria. METHODS AND RESULTS: Two-hundred and thirty carcasses were swabbed and analysed by classical microbiological methods for total E. coli counts and for the presence of pathogenic E. coli. The E. coli counts were high, with a 75th percentile of 444.75 CFUs cm(-2). For pathogenic E. coli, more than 7% of the tested carcasses were positive for E. coli O157. Eighteen E. coli O157 strains were isolated and typed by multiplex PCR. The main isolated pathotype (78%) was eae+ stx2+ ehxA+. In addition to E. coli O157, other attaching and effacing E. coli (AEEC) were also detected from carcasses by colony hybridization after pre-enrichment and plating on sorbitol MacConkey agar using eae, stx1 and stx2 probes. Thirty carcasses (13%) on the 230 analysed harboured at least one colony positive for one of the tested probes. These positive carcasses were different from those positive for E. coli O157. Sixty-six colonies (2.9%) positive by colony hybridization were isolated. The majority (60.6%) of the positive strains harboured an enteropathogenic E. coli-like pathotype (eae+ stx-). Only three enterohaemorrhagic E. coli (EHEC)-like (eae+ stx1+) colonies were isolated from the same carcass. These strains did not belong to classical EHEC serotypes. CONCLUSIONS: In this study, the global hygiene of the slaughterhouse was low, as indicated by the high level of E. coli count. The prevalence of both E. coli O157 and other AEEC was also high, representing a real hazard for consumers. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study of this type in Algeria, which indicates that the general hygiene of the slaughterhouse must be improved.

Abattoirs↗
PubMed2005

Development of a genetic traceability test in pig based on single nucleotide polymorphism detection.

In order to assure traceability along the meat transformation process, a powerful system is required. The administrative traceability shows limits that the use of genetic markers could overcome. The individual genomes contain sequence differences, basis of the genetic polymorphism of which the genetic markers are the witnesses. Among them, two classes seem to dominate on the traceability field: the microsatellites and the single nucleotide polymorphisms (SNP). The aim of this work was to develop a genetic traceability test in pig based on SNPs mainly located in 5' and 3' untranslated regions (UTRs). A set of 21 SNP markers including new SNPs identified in this study and SNPs previously described was selected. A genotyping assay was performed on 96 individuals representing the major crossbred of the pig population in Belgium. Results showed that all individuals tested presented a different genotype. This genotyping method might help the administrative system to guarantee the traceability of pork meat along the transformation process.

Animal Identification Systems↗
PubMed2005

A PCR method for detection of bifidobacteria in raw milk and raw milk cheese: comparison with culture-based methods.

Bifidobacteria are well known for their beneficial effects on health and are used as probiotics in food and pharmaceutical products. As they form one of the most important groups in both human and animal feces, their use as fecal indicator organisms in raw milk products has recently been proposed. Bifidobacteria species isolated in humans are different from those isolated in animals. It should therefore be possible to determine contamination origin (human or animal). A method of detecting the Bifidobacterium genus was developed by PCR targeting the hsp60 gene. The genus Bifidobacterium was identified by PCR amplification of a 217-bp hsp60 gene fragment. The degenerated primer pair specific to the Bifidobacterium genus used was tested for it specificity on 127 strains. Sensitivity was measured on artificially contaminated samples. Food can however be a difficult matrix for PCR testing since it contains PCR inhibitors. So an internal PCR control was used. An artificially created DNA fragment of 315 bp was constructed. The PCR detection method was tested on raw milk and cheese samples and compared with three culture-based methods, which comprised enrichment and isolation steps. The enrichment step used Brain Heart Infusion medium with propionic acid, iron citrate, yeast extract, supplemented with mupirocin (BHMup) or not (BH) and the isolation step used Columbia blood agar medium, supplemented with mupirocin (CMup) or not (C). The method using mupirocin at both enrichment and isolation steps and the PCR method performed from the culture in BHMup enrichment medium were shown to be the most efficient. No significant difference was observed in raw milk samples between PCR from BHMup and the culture-based method BHMup/CMup, while a significant difference was noticed between the same methods in raw milk cheese samples, which would favor using PCR. The results suggested that PCR on the hsp60 gene was convenient for a rapid detection of bifidobacteria in raw milk and raw milk cheese samples and that bifidobacteria always present throughout raw milk cheese production could be efficiently used as fecal indicators.

Animals↗
PubMed2005

Survey of the contamination of foodstuffs of animal origin by Shiga toxin producing Escherichia coli serotype O157:H7 in Belgium from 1999 to 2003.

A survey of the prevalence of Shiga toxin-producing Escherichia coli (STEC) of O157 serotype in foodstuffs of animal origin (beef, veal, pork, chicken, fish) from 1999 to 2003 in Belgium was performed. STEC strains were only isolated from beef with a prevalence of 0.73%. This percentage is low in comparison with the prevalence in other countries. Among the 76 isolated STEC O157 strains, 75% belonged to the serotype O157:H7 and 25% to the serotype O157 non H7. Moreover, the most frequent pathotype was eae stx2 ehxA (74%).

Animals↗
PubMed2004

Comparison of four different methods for Salmonella detection in fecal samples of porcine origin.

Performances of four detection methods were evaluated for recovery of Salmonella spp. in naturally contaminated fecal specimens of porcine origin. The NMKL 71 method consisted of enrichment in Rappaport-Vassiliadis broth and plating on xylose-lysine-desoxycholate medium, whereas the SP-VG-M002 method relied on a Diasalm enrichment followed by streaking on xylose-lysine-tergitol 4 agar (XLT-4). The VIDAS SLM method was composed of double enrichment in Muller-Kauffmann tetrathionate broth and in M broths before processing in a VIDAS device. If the results were positive, the VIDAS ICS immunoenrichment was performed and the result transferred onto three different selective media. The VIDAS ICS protocol is an immunoconcentration step followed by plating on XLT-4. Seventy-eight samples were tested with all four methods simultaneously, leading to 34 positive samples with at least one method. For this assay, VIDAS SLM revealed 31 positive samples (91.2%), whereas the average positive percentage of the three other methods was 37.3% (P < 0.001). Two-paired comparisons with the VIDAS SLM method were also performed. McNemar values were systematically highly significant (P < 0.001). The proportion of agreement was significantly inferior (P < 0.05) for the comparison of VIDAS ICS and VIDAS SLM (68.7%) compared with the two other paired comparisons (average percentage, 81.5%). The conclusion reached by this trial is that VIDAS SLM significantly improves the recovery of Salmonella in naturally contaminated fecal specimens. For the paired-comparisons, NMKL 71 and SP-VG-M002 were comparable in terms of efficiency, whereas the VIDAS ICS protocol, as established by the manufacturer for food samples only, seemed less efficient than the other two.

Animals↗
PubMed2004

Discrimination between Bifidobacterium species from human and animal origin by PCR-restriction fragment length polymorphism.

Bifidobacteria are normal intestinal flora in humans and animals. The genus Bifidobacterium includes 31 species of significant host specificity. Taking into account their properties, we proposed to use bifidobacteria as fecal contamination indicators. PCR-restriction fragment length polymorphism on the 16S rDNA gene was used to distinguish the different Bifidobacterium species. Sixty-four strains belonging to 13 different species were differentiated from animal or human origin using one or two restriction enzymes. Moreover, the primers used were specifics of the Bifidobacterium genus. Therefore, this method made it possible to determine both the presence of bifidobacteria in a sample and its origin of contamination.

Animals↗
PubMed2002

A role for the Clostridium perfringens beta2 toxin in bovine enterotoxaemia?

Non-enterotoxigenic type A Clostridium perfringens are associated with bovine enterotoxaemia, but the alpha toxin is not regarded as responsible for the production of typical lesions of necrotic and haemorrhagic enteritis. The purpose of this study was to investigate the putative role of the more recently described beta2 toxin. Seven hundred and fourteen non-enterotoxigenic type A C. perfringens isolated from 133 calves with lesions of enterotoxaemia and high clostridial cell counts (study population) and 386 isolated from a control population of 87 calves were tested by a colony hybridisation assay for the beta2 toxin. Two hundred and eighteen (31%) C. perfringens isolated from 83 calves (62%) of the study population and 113 (29%) C. perfringens isolated from 51 calves (59%) of the control population tested positive with the beta2 probe. Pure and mixed cultures of four C. perfringens (one alpha+beta2+, one alpha+enterotoxin+ and two alpha+) were tested in the ligated loop assay in one calf. Macroscopic haemorrhages of the intestinal wall, necrosis and haemorrhages of the intestinal content, and microscopic lesions of necrosis and polymorphonuclear and mononuclear cell infiltration of the intestinal villi were more pronounced in loops inoculated with the alpha and beta2-toxigenic C. perfringens isolate. These results suggest in vivo synergistic role of the alpha and beta2 toxins in the production of necrotic and haemorrhagic lesions of the small intestine in cases of bovine enterotoxaemia. However, isolation of beta2-toxigenic C. perfringens does not confirm the clinical diagnosis of bovine enterotoxaemia and a clostridial cell counts must still be performed.

Animals↗
PubMed2001

Bacterial intestinal flora associated with enterotoxaemia in Belgian Blue calves.

The enterotoxaemia syndrome in Belgian Blue calves is characterised by a high case fatality rate, sudden death, lesions of haemorrhagic enteritis of the small intestine and, quite often an absence of other clinical signs but its cause has not been yet identified. As a first step in this identification, the aerobic and anaerobic intestinal flora of a population of 78 calves, originating from farms located in southern Belgium and that died in circumstances defined as "calf enterotoxaemia" (study population) and of 64 calves that died in other circumstances (control population) were studied qualitatively and quantitatively. The colonies were identified after subcultures with appropriate API sugar sets. Anaerobically Clostridium perfringens was isolated in higher numbers (mean values of 10(7)-10(7.5) colony forming units (CFU) versus 10(4)-10(5) CFU per ml of intestinal content) and from more animals (79 versus 19%) in the study population than in the control population, although individual results from both populations could overlap. Other clostridial species, i.e. mainly urease-negative C. sordellii and C. bifermentans, were isolated in high numbers (>10(6) CFU per ml of intestinal content) from a few animals in the study population only. All but one of the 705 C. perfringens isolates from both populations belonged to the A toxin type and none of the urease-negative C. sordellii was toxigenic. Gram-negative anaerobes were not isolated in high numbers from any of the samples. Aerobically beta-haemolytic E. coli were significantly more frequent among the study population, but were isolated from only 25% of the animals. Salmonella Typhimurium was isolated from only two animals in the study population. Less than 1% of the E. coli isolated were verotoxigenic and one-third were necrotoxigenic. At this stage only non-enterotoxigenic type A C. perfringens are thus statistically associated with the enterotoxaemia syndrome in Belgian Blue calves and fulfil the first of the Koch's postulates.

Animals↗
PubMed2001

Antimicrobial susceptibilities of Campylobacter strains isolated from food animals in Belgium.

Campylobacter spp. are a frequent cause of diarrhoea in man, originating mostly from poultry. It has been suggested that the veterinary use of antibiotics is largely responsible for resistance in human isolates, particularly to quinolones. During a 6 month period from June to December 1998, 677 Campylobacter isolates were obtained from healthy poultry and pigs. Samples were taken at Belgian slaughterhouses. Species identification was performed by biochemical tests, multiplex PCR and SDS-PAGE of whole-cell proteins. The in vitro susceptibility to six antimicrobial drugs was determined by the agar dilution method. Campylobacter jejuni was found more often in poultry than Campylobacter coli (79% C. jejuni versus 21% C. coli). In pigs the situation was reversed (6 versus 94%). Erythromycin resistance was significantly higher (P < 0.05) in C. coli, particularly in C. coli isolated from pigs (67.2%). Alarmingly high rates of resistance to ciprofloxacin were also noted, particularly for C. coli from broilers (62.1%). The latter indicates that resistance of Campylobacter in humans could derive from animals.

Animals↗
PubMed1999

[Study of the role of Clostridium perfringens in bovine enterotoxemia].

Bovine enterotoxaemia is an acute to peracute syndrome occurring mainly in calves and characterized by the sudden or very rapid death of the calf, with colics, convulsions and nervous disorders as clinical signs, if any. The most pronounced lesion is a necrohaemorrhagic enteritis of the jejunum, the ileum, and sometimes the colon. Suckling beef calves are the most frequently affected ones. In 67% of the 78 field cases investigated, some kind of stress was observed 24 to 36 hours prior to the death: change in diet or pasture, vaccination... The most frequently isolated bacteria, and the one isolated in highest numbers, was non-sporulated non-enterotoxigenic toxinotype A Clostridium perfringens. Reproduction of the lesions was successful in a ligated intestinal loop assay in one calf with a few of these strains, more especially with one of them, which was shown later to produce another recently described toxin, the beta 2 toxin. A role for this beta 2 toxin in bovine enterotoxaemia is thus speculated for future research.

Animals↗
PubMed1998

Virulence plasmids of enterotoxigenic Escherichia coli isolates from piglets.

Virulence plasmids of 68 ETEC isolates from piglets belonging to different pathotypes and six ETEC isolates from calves with pathotypes typical of porcine ETEC were identified with seven virulence probes for the heat-stable (STa and STb) and heat-labile (LT) enterotoxins, for the F4, F5, F6, and F41 fimbrial adhesion subunit, and also with five Rep probes for the RepFIA and RepFIB basic replicons, and the RepFIC family of basic replicons. With the exception of the F41 probe, the other virulence probes hybridized with at least one plasmid band of a size range from 65 to more than 100 Mda. Common associations of virulence factor-encoding genes on plasmid bands were: STb/LT, STa/F5, STa/F6, STa/STb. Other associations, STa/F4, STa/F4/F6, and STa/STb/LT/F6, were rarer. On the other hand the F4 adhesin-encoding genes were isolated on one plasmid band in all but three F4+ isolates. All but one of the 92 virulence plasmids which were studied have Rep probe hybridization profiles and replicon types typical of the uni- or multireplicon plasmids belonging to the various incompatibility groups of the F incompatibility complex.

Animals↗
PubMed1998

An efficient sampling technique used to detect four foodborne pathogens on pork and beef carcasses in nine Belgian abattoirs.

The method presented in this paper should prove useful in assessing the effectiveness of HACCP plans developed in slaughterhouses. Samples were collected by swabbing well-defined areas of pork and beef carcasses with sterile gauze. Between 160 and 420 half-carcasses were swabbed in each of nine pork or beef slaughterhouses. Swabs from five carcasses were placed in the same sterile Stomacher bag, constituting a single composite sample. Standard or validated analytical methods were used to isolate and characterize four foodborne pathogens. Salmonella spp., Listeria monocytogenes, Campylobacter spp., and verocytotoxin-producing E. coli were detected, respectively, in 27, 2, 2, and 14% of the pork samples and 0, 22, 10, and 5% of the beef samples. Of the 10 samples positive for E. coli O157, only one yielded an isolate confirmed to be enterohemorrhagic. Since Salmonella spp. appear as the main contaminant port (27%) and L. monocytogenes as the main containment of beef (22%), any slaughterhouse sampling plan should include testing for the former in the case of pork carcasses and for the latter in the case of beef carcasses. One should also test regularly for the presence of E. coli O157 and Campylobacter spp. in pork and beef abattoirs. The method presented here is an easy way to assess the contamination rate of carcasses at the end of the slaughtering process.

Abattoirs↗
PubMed1997

Clostridium perfringens urease genes are plasmid borne.

Although many bacteria are ureolytic, and in some cases urease acts as a virulence factor, the urease phenotype has not been analyzed in the anaerobic pathogen Clostridium perfringens. In this study, approximately 2% of C. perfringens strains, representing the principal biotypes, were found to harbor the urease structural genes, ureABC, and these were localized on large plasmids that often encode, in addition, the lethal epsilon or iota toxins or the enterotoxin. This represents the first report of a plasmid-encoded urease in a gram-positive bacterium. The C. perfringens enzyme was highly similar to the ureases of other bacteria and cross-reacted with antibodies raised against the urease purified from Helicobacter pylori. Urease production was inhibited by urea and induced under growth conditions where the availability of nitrogen sources was limiting. To date, this form of regulation has been observed only for chromosomal ureABC genes.

Amino Acid Sequence↗
PubMed1996

Genome mapping of Clostridium perfringens strains with I-CeuI shows many virulence genes to be plasmid-borne.

The intron-encoded endonuclease I-CeuI from Chlamydomonas eugametos was shown to cleave the circular chromosomes of all Clostridium perfringens strains examined at single sites in the rRNA operons, thereby generating ten fragments suitable for the rapid mapping of virulence genes by pulsed-field gel electrophoresis (PFGE). This method easily distinguishes between plasmid and chromosomal localisations, as I-CeuI only cuts chromosomal DNA. Using this approach, the genes for three of the four typing toxins, beta, epsilon, and tau, in addition to the enterotoxin and lambda-toxin genes, were shown to be plasmid-borne. In a minority of strains, associated with food poisoning, where the enterotoxin toxin gene was located on the chromosome, genes for two of the minor toxins, theta and mu, were missing.

Bacterial Toxins↗
PubMed1996

Hybridization of 2,659 Clostridium perfringens isolates with gene probes for seven toxins (alpha, beta, epsilon, iota, theta, mu, and enterotoxin) and for sialidase.

OBJECTIVE: To genetically characterize Clostridium perfringens isolates for association of pathologic type with various diseases. DESIGN: Prospective study. SAMPLE POPULATION: 2,659 C perfringens isolates from various nonhuman animals species, human beings, and foods. PROCEDURE: Colony hybridization with DNA probes for 7 toxin (alpha, beta, epsilon, iota (subunits a and b), theta, mu, and enterotoxin) genes and 1 sialidase gene were performed to group the isolates by pathologic type. RESULTS: Enterotoxin-negative type-A isolates were the most common (2,575/2,659), were isolated from all sources, and were separated into 5 pathologic types. In cattle and horses with enterotoxemia, essentially only these pathologic types were identified. The enterotoxin-negative isolates of types C or D each had a single pathologic type. Type-C isolates were isolated only from swine with necrotic enteritis and type-D isolates from small ruminants with enterotoxemia, except that 1 type-D isolate was also found from a healthy fish. Type-B or -E isolates were not found. Among the 47 enterotoxin-positive isolates, 5 isolates from sheep or deer were type D and the other 42 were type A. These 42 isolates were grouped into 3 pathologic types: 1 type was isolated from samples of almost all origins, but the other 2 types were found in only 5 fish, 4 human beings, and 1 dog. CONCLUSIONS AND CLINICAL RELEVANCE: Genetic characterization of these isolates allowed identification of 11 different pathologic types. This approach may be useful in molecular diagnosis and prophylaxis of clostridial disease.

Animals↗
PubMed1995

The enterotoxin gene (cpe) of Clostridium perfringens can be chromosomal or plasmid-borne.

The location of the cpe gene, encoding the enterotoxin responsible for food poisoning in humans, has been studied in a series of enterotoxigenic Clostridium perfringens strains by means of pulsed field gel electrophoresis of genomic DNA. The cpe gene was found at the same chromosomal locus in strains associated with food poisoning in humans and was shown to be linked to a repetitive sequence, the HindIII repeat, and an open reading frame, ORF3, that may be part of an insertion sequence. In contrast, when the strains originated from domesticated livestock cpe was located on a large episome where it was often close to a copy of the transposable element IS1151. In these cases, the HindIII repeat was not linked to the cpe gene although this was generally preceded by ORF3.

Blotting, Southern↗