Decrease of Salmonella and Escherichia coli O157:H7 counts during dry-aging of beef but potential growth of Listeria monocytogenes under certain dry-aging conditions.

Dry-aging of beef comprises the storage of carcasses and (sub)primal cuts at a low temperature and relative humidity for a prolonged period, aiming to increase the sensory quality of meat. Limited data are available on the survival and potential growth of pathogens on the surface of beef during dry-aging. Therefore, this study evaluates the changes in Salmonella, Shiga toxin-producing Escherichia coli O157:H7 and Listeria monocytogenes counts during dry-aging. A mixture of pathogenic strains was inoculated on the surface of beef loins, which were stored under four different process conditions (2 °C and 6 °C × relative humidity 75 and 85% during 42 days). Salmonella and E. coli O157:H7 counts significantly decreased during dry-aging. The daily reductions varied from -0.07 to -0.14 log10 CFU and from -0.09 to -0.14 log10 CFU, respectively, depending on the loin, matrix and condition. The reduction of L. monocytogenes was slower, with a maximum of -0.07 log10 CFU/day. L. monocytogenes counts increased with 1.0 log10 CFU on the lean meat of one loin with pH > 6.0 at the end of dry-aging, indicating that this pathogen can potentially grow under certain dry-aging conditions.

Influence of aging time, temperature and relative humidity on the sensory quality of dry-aged Belgian Blue beef.

The objective of this study was to evaluate the effect of aging period (0, 3, 6 or 9 weeks), aging temperature (2 versus 6 °C at 75% relative humidity, experiment 1) and relative humidity (70 versus 90% at 2 °C, experiment 2) on the sensory traits, oxidative stability and proteolysis of Belgian Blue beef. For each experiment, eight loins (M. longissimus thoracis et lumborum) from four animals (left and right side) were assigned to one of the two treatments (n = 4). Results showed no further tenderization after three weeks of aging, whereas metmyoglobin formation and lipid oxidation increased until nine weeks of aging (P < 0.05). During the nine weeks of aging, atypical flavor, odor and flavor intensity was affected (P < 0.05). This was accompanied by an increase of small peptides and other nitrogenous compounds. Aging temperature and relative humidity had only a very limited effect on the quality traits.

Salmonella prevalence and persistence in industrialized poultry slaughterhouses.

Salmonella contamination sources and transmission routes were studied in 5 Belgian poultry slaughterhouses. Samples from the slaughter and cutting line after cleaning and disinfection were collected, as well as neck skin samples and thighs during slaughter of the first flock. In total, 680 swab and water samples were taken from the slaughter line before slaughter. In all slaughterhouses, Salmonella was notwithstanding cleaning and disinfection still isolated from the slaughter line before start of activities. The prevalence of Salmonella in the plucking area was 10.4% (38/365) (hanging area: 5.0%, scalding tank: 5.8%, plucking machine: 17.0%); in the evisceration room, 1.5% (2/138); and in the cutting area, 2.0% (3/149). No Salmonella (0/28) was found in samples from the chilling line. On neck skin samples taken from the various lines, Salmonella prevalence was 16.1% (48/299) after plucking, 16.0% (48/300) after evisceration, 23.3% (70/300) after chilling; on thighs, prevalence was 10.0% (24/240). Nine Salmonella serotypes were identified of which Salmonella Infantis was the most common serovar (53.8%), especially in slaughterhouse A. Two contamination causes were identified; first, although all flocks had an official Salmonella negative status, this was in one case incorrect and led to an enormous contamination of the neck skins of the flock and the slaughterline (i.e., cooling water). Second, molecular typing revealed cross-contamination from flocks slaughtered 1 d before sampling. Salmonella was apparently not always eliminated by the cleaning and disinfection process and able to contaminate the carcasses of the first slaughtered flock. In conclusion, the results of this study provided practical insights for poultry production to further improve their Salmonella control, for example, Salmonella status determination closer to the slaughter date, to adapt cleaning and disinfection protocols especially for critical machinery and better hygienic designed equipment.

The effect of partial depopulation on Campylobacter introduction in broiler houses.

Poultry is seen as the main reservoir for Campylobacter. Control of this zoonotic pathogen in primary production could potentially reduce the colonization in broiler flocks and consequently reduce the number of human infections. In the present study, 20 broiler flocks from 10 farms, were sampled immediately before and 5 to 7 d after partial depopulation (thinning) for the presence of Campylobacter using cecal droppings and overshoes. At the time of thinning, the catching crew, transportation vehicles, forklift, and transport containers were sampled for the presence of Campylobacter. Samples were cultivated; presumed positive isolates were confirmed by PCR. The isolates were molecularly typed by flaA restriction analysis and pulsed field gel electrophoresis. Results show that all flocks were thinned using Campylobacter-contaminated equipment and materials. One-third of the broiler flocks became colonized after thinning. In 67% of the colonization cases, identical strains were found matching those of container systems, transport trucks, and/or forklifts. This identifies thinning as an important risk factor for Campylobacter introduction into broiler houses. Setup and compliance with biosecurity practices during thinning is essential to prevent Campylobacter colonization of broiler flocks.

Occurrence of 'gang of five' Shiga toxin-producing Escherichia coli serogroups on Belgian dairy cattle farms by overshoe sampling.

Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens responsible for global outbreaks. This study was conducted to investigate the occurrence of 'gang of five' STEC serogroups (O26, O103, O111, O145, O157) on Belgian dairy cattle farms by overshoe (OVS) sampling, and to evaluate the presence of virulence genes in the obtained isolates. A total of 88 OVS, collected from the pen beddings of 19 Belgian dairy cattle farms, were selectively enriched in mTSBn, followed by immunomagnetic separation and plating onto CT-SMAC for O157 STEC isolation, as well as in Brila broth, followed by a selective acid treatment and plating onto CHROMagarTM STEC and chromIDTM EHEC for non-O157 STEC isolation. Overall, 11 of 19 farms (58%) tested positive for presence of 'gang of five' STEC. O26 STEC was most frequently isolated from OVS (11/88; 12·5%), followed by O157 (10/88; 11·5%), O145 (3/88; 3·5%) and O103 (3/88; 3·5%). Additionally, 35% of the OVS collected from pens housing young cattle 1-24 months of age tested positive for 'gang of five' STEC, indicating that this age category is more likely to harbour STEC compared to new-born and adult cattle. Importantly, half of the obtained 'gang of five' STEC isolates (48%) possessed the eae and stx2 gene, suggesting a high pathogenic potential to humans.

Pathogenic potential of Escherichia coli O157 and O26 isolated from young Belgian dairy calves by recto-anal mucosal swab culturing.

AIMS: The purpose of this study was to investigate the occurrence of Escherichia coli O157 and O26 on Belgian dairy cattle farms, the presence of virulence genes in the confirmed isolates and the association of E. coli O26 presence with calf diarrhoea. METHODS AND RESULTS: In total, 233 recto-anal mucosal swabs (RAMS) were obtained from healthy and diarrheic dairy calves on three farms, each alternately visited three consecutive times. RAMS were analysed for presence of E. coli O157 and O26, and stx1, stx2 and eae virulence genes. Overall, 19% of RAMS tested positive for E. coli O157, while 31% tested positive for E. coli O26. The majority of isolates possessed both stx and eae, denoting a high pathogenic potential to humans. While both serogroups persisted at farm level, persistence within the same animal over time appeared to be relatively rare. Interestingly, E. coli O26 was already abundantly present at a younger age compared to E. coli O157. Calf diarrhoea could not be associated with presence of E. coli O26. CONCLUSIONS: Young dairy calves are important on-farm reservoirs of potentially pathogenic E. coli O157 and O26. A role of E. coli O26 in calf diarrhoea could not be confirmed. SIGNIFICANCE AND IMPACT OF THE STUDY: O157 and O26 are responsible for the majority of human STEC infections. Gaining more epidemiological information regarding their occurrence and persistence on cattle farms will contribute to a better understanding of STEC ecology and risk of human transmission.

Occurrence, distribution and diversity of Listeria monocytogenes contamination on beef and pig carcasses after slaughter.

In this study we investigated the prevalence and location of Listeria monocytogenes and hygiene indicator bacteria on beef and pig carcasses. Carcasses were sampled after slaughter and before cooling at eight and nine sites on the carcass, respectively. For each sample, detection and enumeration of Listeria was performed, as well as the enumeration of Total Aerobic Counts (TAC) and Enterobacteriaceae. The L. monocytogenes isolates were also typed to determine pulsotypes and clonal complexes (CC). L. monocytogenes was detected on 46% [95% CI: 35-56%] of beef and 22% [95% CI: 11-32%] of pig carcasses. Contamination levels at the different carcass sites differed considerably between beef and pigs. Genetic typing of strains suggests that carcass contamination originates from both incoming animals with transmission during slaughter practices as well as persistent (CC9) contamination from the slaughterhouse environment. These findings can be used to understand the complexity of introduction and persistence of this pathogen in slaughter facilities. Accurate correlation of L. monocytogenes presence proved unfeasible with any of the tested hygiene indicator bacteria.

Relation between broiler and human Campylobacter jejuni strains isolated in Belgium from 2011 to 2013.

AIMS: This study inquires the relationship between Campylobacter jejuni isolated from broiler meat carcasses (n = 97) and human clinical samples (n = 72) in Belgium, from 2011 to 2013. METHODS AND RESULTS: The evaluation of the relation was based on the characteristics determined using multilocus sequence typing (MLST) alone and combined with flagellin gene A restriction fragment length polymorphism (flaA-RFLP) typing, antibiotic microbiological resistance profiling (AMRp), lipooligosaccharide class typing or virulence gene profiling (Vp). Clusters containing both human and broiler meat strains were more common when MLST was used alone, followed by MLST/flaA-RFLP and then by MLST/AMRp. More logical chronologically relations broiler-human were obtained for MLST/flaA-RFLP, then for MLST, and finally for MLST/AMRp: i.e. the isolates would first be detected in the broiler meat and at the same time or later in humans. CONCLUSIONS: In several cases, the C. jejuni strains isolated from the consumed broiler meat and from the campylobacteriosis case had the same profile, according to the used typing methods. The circulating Campylobacter strains appear to have remained the same from 2011 till 2013 in Belgium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study corroborates previously published data from Belgium that suggest a strong correlation between C. jejuni strains isolated from broiler meat and from campylobacteriosis patients.

Reduced contamination of pig carcasses using an alternative pluck set removal procedure during slaughter.

This study compared the current pig slaughter procedure where the pluck set is completely removed with a procedure where the pluck set is partially removed, leaving the highly contaminated oral cavity, tonsils and tongue untouched. The effect on carcass contamination was investigated by enumerating hygiene indicator bacteria (total aerobic count, Enterobacteriaceae and E. coli) and cefotaxime-resistant E. coli (CREC) as well as assessing Salmonella and Yersinia enterocolitica presence on the sternum, elbow and throat of pig carcasses. Using the alternative pluck set removal, significantly lower mean numbers of hygiene indicator bacteria on throat samples and E. coli on elbow samples were found. Less pig carcasses were highly contaminated and a lower presence and level of CREC was observed. No difference in Salmonella or Yersinia enterocolitica presence was seen. The data in this study can help to assess the effect of this alternative procedure on the safety of pork and subsequently public health.

Quantification, distribution and diversity of ESBL/AmpC-producing Escherichia coli on freshly slaughtered pig carcasses.

This study quantified cefotaxime-resistant E. coli (CREC) on nine different carcass areas of 104 freshly slaughtered pig carcasses. In 49% [95% confidence interval (95% CI): 29-69%] of the carcasses CREC could be isolated and enumerated (using Tryptone Bile Agar with X-Glucuronide supplemented with 1 mg/L cefotaxime). Proportions of positive samples varied between carcass areas from 1% [95% CI: 0-10%] (loin) to 23% [95% CI: 10-44%] (head). Maximum concentrations on positive samples ranged between -0.6 log10 CFU/cm2 (loin, elbow before evisceration) and 1.7 log10 CFU/cm2 (head). The head was significantly more frequently contaminated than the loin (p = 0.027) and ham (3% [95% CI: 1-15%]). The foreleg was significantly more frequently contaminated (20% [95% CI: 13-30%]) than the ham. Combination disk diffusion assays revealed that 81% of the CREC isolates were extended-spectrum beta-lactamases (ESBL) producers, 13% were AmpC cephalosporinases (AmpC) producers and 2% ESBL and AmpC co-producers. Genotyping denoted blaCTX-M-gr1 (63%) and blaTEM (40%) as most present antibiotic resistance genes. Multiple gene combinations in one isolate and multiple combinations of genotypes and phenotypes among isolates of one sample were observed. These quantitative data can be used for intervention strategies to lower human exposure to CREC.

Correlation between slaughter practices and the distribution of Salmonella and hygiene indicator bacteria on pig carcasses during slaughter.

This study investigated the distribution of hygiene indicator bacteria and Salmonella on pig carcasses. Moreover, the relation between hygiene indicator counts and Salmonella presence as well as associations between specific slaughter practices and carcass contamination were determined for each carcass area. Seven Belgian pig slaughterhouses were visited three times to swab five randomly selected carcasses at nine different areas, after evisceration and trimming. Information about slaughter practices was collected using a questionaire. In all samples, the E. coli and Salmonella presence was analyzed and Enterobacteriaceae and total aerobic bacteria were quantified. Average total aerobic counts ranged from 3.1 (loin, pelvic duct, ham) to 4.4 log10 CFU/cm2 (foreleg). Median Enterobacteriaceae numbers varied between 0.4 (ham) an 1.8 log10 CFU/cm2 (foreleg). E. coli and Salmonella presence ranged from 15% (elbow) to 89% (foreleg) and 5% (elbow) to 38% (foreleg), respectively. Positive relations were found between hygiene indicator counts and Salmonella presence at the head, sternum, loin and throat. Several slaughter practices, such as splitting the head and incising tonsils, were associated with higher levels of hygiene indicator bacteria and Salmonella. These findings can be used to educate slaughterhouse personnel and estimate the public health risk involved in consumption of different pork cuts.

High abundance and diversity of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in faeces and tonsils of pigs at slaughter.

This cross-sectional study investigates the abundance of cefotaxime-resistant Escherichia coli (CREC) in the faeces and tonsils of 96 pigs during slaughter. Moreover, different isolates from a selected number of pigs were tested to study the diversity of blaESBL genes within E. coli isolates from one pig. Cefotaxime-resistant bacteria (based on enumeration results on MacConkey agar supplemented with 1mg/L cefotaxime) were found in the faeces of 77 pigs (80%; 95% CI: 70-87%) and the tonsils of 91 pigs (95%; 95% CI: 88%-98%). Cefotaxime-resistant E. coli (based on enumeration results on Tryptone Bile X-glucuronide agar supplemented with 1mg/L cefotaxime) were detected in 72 faecal samples (75%; 95% CI: 64-83%) and 45 tonsil samples (47%; 95% CI: 35-59%), in numbers up to 5.5 and 5.6log10 CFU/g, respectively. On average, around 1/10,000 E. coli in both faeces and tonsils were cefotaxime-resistant, though large variations were observed between pigs. Within one sample, CREC isolates with up to five different combinations of ESBL genes were observed. In three out of 16 faecal samples and six out of 14 tonsil samples, only one ESBL gene profile was found. The high numbers of CREC that are occasionally found in the faeces and tonsils of pigs during slaughter may represent an important source of contamination of carcasses and subsequently pork.

Control of human pathogenic Yersinia enterocolitica in minced meat: Comparative analysis of different interventions using a risk assessment approach.

This study aimed to evaluate the effect of different processing scenarios along the farm-to-fork chain on the contamination of minced pork with human pathogenic Y. enterocolitica. A modular process risk model (MPRM) was used to perform the assessment of the concentrations of pathogenic Y. enterocolitica in minced meat produced in industrial meat processing plants. The model described the production of minced pork starting from the contamination of pig carcasses with pathogenic Y. enterocolitica just before chilling. The endpoints of the assessment were (i) the proportion of 0.5 kg minced meat packages that contained pathogenic Y. enterocolitica and (ii) the proportion of 0.5 kg minced meat packages that contained more than 10³ pathogenic Y. enterocolitica at the end of storage, just before consumption of raw pork or preparation. Comparing alternative scenarios to the baseline model showed that the initial contamination and different decontamination procedures of carcasses have an important effect on the proportion of highly contaminated minced meat packages at the end of storage. The addition of pork cheeks and minimal quantities of tonsillar tissue into minced meat also had a large effect on the endpoint estimate. Finally, storage time and temperature at consumer level strongly influenced the number of highly contaminated packages.

Assessment of factors influencing the within-batch seroprevalence of human enteropathogenic Yersinia spp. of pigs at slaughter age and the analogy with microbiology.

The microbiologically and serologically-based prevalence of human enteropathogenic Yersinia spp. at moment of slaughter varies between pig farms due to different herd-level factors. A face-to-face questionnaire concerning a broad range of farm aspects (e.g., management and housing system, biosecurity, and hygiene measurements) was performed on one hundred farms. Factors influencing the seropositivity of 7047 pigs against human pathogenic Yersinia spp. were determined and compared to the microbiology. At the slaughterhouse, pieces of diafragm of on average 70 slaughter pigs per batch were sampled to determine the level of antibodies against enteropathogenic Yersinia spp. After univariable mixed-effect logistic regressions, variables that were related to the seropositivity (p<0.05) were included in a multivariable model (p<0.1). The factors remaining significantly associated in the latter model were an increasing number of piglet suppliers (zero up to eleven suppliers) (Odds Ratio=1.4), a high density of pig farms in the area (high versus low density) (Odds Ratio=2.3), the use of semislatted floors in the fattening pig unit (semi slatted floor versus fully slatted floor) (Odds Ratio=3.8) and the possibility of snout contact in the fattening pig unit (snout contact or not) (Odds Ratio=0.1). Decreasing the risk of infection with human enteropathogenic Yersinia spp. at moment of slaughter or during rearing is possible by changing farm management factors.

Comparative performance of isolation methods using Preston broth, Bolton broth and their modifications for the detection of Campylobacter spp. from naturally contaminated fresh and frozen raw poultry meat.

The performance of different isolation methods was evaluated for the detection of Campylobacter from naturally contaminated raw poultry meat. Therefore, fresh and frozen poultry meat samples were analysed using the standard procedure (ISO 10272-1:2006), enrichment in Preston broth, and enrichment in modified Bolton broth (supplemented with (i) potassium clavulanate (C-BB), (ii) triclosan (T-BB), (iii) polymyxin B (P-BB)). The enrichment cultures were streaked onto both modified charcoal cefoperazone deoxycholate agar (mCCDA) and RAPID'Campylobacter agar (RCA). Moreover, direct plating on mCCDA and RCA was performed to quantify Campylobacter. In total, 33 out of 59 fresh retail meat samples (55.9%) were Campylobacter positive. For both fresh and frozen poultry meat samples, enrichment in Bolton broth (ISO 10272-1:2006) resulted in a higher number of positive samples than enrichment in Preston broth. Supplementation of Bolton broth with potassium clavulanate (C-BB) and triclosan (T-BB) enhanced the Campylobacter recovery from fresh poultry meat compared to non-supplemented Bolton broth, although the use of C-BB was less applicable than T-BB for Campylobacter recovery from frozen samples. Additionally, the use of RCA resulted in a higher isolation rate compared to mCCDA. The present study demonstrates the impact of culture medium on the recovery of Campylobacter from fresh and frozen naturally contaminated poultry meat samples and can support laboratories in choosing the most appropriate culturing method to detect Campylobacter.

Contamination of freshly slaughtered pig carcasses with enteropathogenic Yersinia spp.: Distribution, quantification and identification of risk factors.

A cross-sectional survey was undertaken to determine the overall prevalence of enteropathogenic Yersinia spp. in the tonsils, feces and on carcasses of pigs at slaughter. Moreover, factors associated with Yersinia contamination of freshly eviscerated pig carcasses were studied. Yersinia enterocolitica serotype O:3 was isolated from the tonsils and feces of 55.3% and 25.6% of pigs, and Y. pseudotuberculosis from 1.4% and 0.6%, respectively. The pathogens were also recovered from 39.7% of carcass surfaces post-evisceration. The highest prevalence was found at the mandibular region (28.9%), followed by the sternal region (16.4%), pelvic duct (7.8%), and split surface near the sacral vertebrae (6.9%). Regarding the quantification of the pathogen, the median concentration of pathogenic Y. enterocolitica was 4.14l og10 CFU/g in tonsils with countable numbers (n=143) and 2.80 log10 CFU/g for fecal samples with countable numbers (n=26). The quantitative load on the carcass surface was generally low as the majority of the carcass samples (97.0%) had Yersinia concentrations below the detection limit of enumeration (<1.30 log10 CFU/100 cm(2)). The initial presence of Y. enterocolitica in the tonsils and/or feces was significantly associated with carcass contamination at all sampled areas. Other risk factors for carcass contamination are the splitting of the head together with the carcass, and incision of the tonsils during removal of the pluck. Small adaptations in slaughter practices and the training of slaughterhouse personnel to respect basic hygienic instructions may diminish carcass contamination with enteropathogenic Yersinia.

Clearance of Escherichia coli O157:H7 infection in calves by rectal administration of bovine lactoferrin.

Enterohemorrhagic Escherichia coli (EHEC) strains, of which E. coli O157:H7 is the best-studied serotype, are an important group of foodborne pathogens causing severe illness in humans worldwide. The main reservoirs for EHEC are ruminants, mostly cattle, which harbor the bacteria in their intestinal tracts without showing clinical symptoms. In this study, we used bovine lactoferrin, a natural occurring bactericidal and immunomodulating protein, as an antibacterial agent against EHEC infection in cattle. Nine 3-month-old Holstein-Friesian calves were experimentally infected with EHEC (strain NCTC12900). Three animals received a daily rectal spray treatment with bovine lactoferrin, three animals received an oral treatment, and three animals served as a control group. Blood samples were collected weekly and fecal samples twice weekly to monitor antibody responses and fecal excretion, respectively. Animals in the rectal group ceased shedding within 26 days of the experimental treatment and remained negative. This beneficial effect of bovine lactoferrin was not observed in the oral group, where animals were still shedding at the time of euthanasia (day 61). All groups developed serum responses, but no clear differences could be observed between the groups. However, the results indicate that the use of bovine lactoferrin as a rectal treatment can be a useful strategy to preclude further transmission of EHEC infections from cattle to humans.

Influence of isolation methods on the occurrence of plasmid-carrying Yersinia enterocolitica serotype O:3 in slaughter pig tonsils, faeces and carcass surface swabs.

Yersinia enterocolitica is an important foodborne pathogen that is primarily transmitted to humans through the consumption of contaminated pork. Different matrices of pigs at slaughter were tested for the presence of human pathogenic types of Y. enterocolitica using direct plating, selective enrichment, and cold enrichment. Y. enterocolitica serotype O:3 was isolated from the tonsils and faeces of 55.3% and 25.6% of pigs, respectively. The pathogen was also recovered from 15.0% of swab samples taken from the carcass surface post-evisceration. Tonsils positive by direct plating revealed an average concentration of 3.99 log10 Y. enterocolitica per gram, whereas the majority of positive faecal and carcass samples were contaminated below the detection limit of enumeration. The relative sensitivity of the methods to recover pathogenic Y. enterocolitica varied among the different matrices. Nevertheless, cold enrichment was significantly more efficient than direct plating and selective enrichment for all three sample matrices. From the 2082 recovered Y. enterocolitica isolates, 1742 (83.7%) harboured the virulence plasmid. Isolates obtained from faeces were more likely to contain the virulence plasmid than isolates from tonsils and carcass swabs. To obtain reliable results regarding the presence of plasmid-carrying Y. enterocolitica isolates, sensitive isolation methods should be combined with testing of a sufficient number of isolates.

Genetic characteristics of Shiga toxin-producing E. coli O157, O26, O103, O111 and O145 isolates from humans, food, and cattle in Belgium.

In this study, we characterized 272 Shiga toxin-producing Escherichia coli (STEC) isolates from humans, food, and cattle in Belgium [O157 (n = 205), O26 (n = 31), O103 (n = 15), O111 (n = 10), O145 (n = 11)] for their virulence profile, whole genome variations and relationships on different genetic levels. Isolates of O157 displayed a wide variation of stx genotypes, heterogeneously distributed among pulsogroups (80% similarity), but with a concordance at the pulsosubgroup level (90% similarity). Of all serogroups evaluated, the presence of eae was conserved, whereas genes encoded on the large plasmid (ehx, espP, katP) occurred in variable combinations in O26, O103, and O145. The odds of having haemolytic uraemic syndrome was less for all genotypes stx2a, stx2c, stx1/stx2c, and stx1 compared to genotype stx2a/stx2c; and for patients aged >5 years compared to patients aged ≤ 5 years. Based on the genetic typing and by using epidemiological data, we could confirm outbreak isolates and suggest epidemiological relationships between some sporadic cases. Undistinguishable pulsotypes or clones with minor genotypic variations were found in humans, food, and cattle in different years, which demonstrated the important role of cattle as a reservoir of STEC O157, and the circulation and persistence of pathogenic clones.

Assessment of human exposure to 3rd generation cephalosporin resistant E. coli (CREC) through consumption of broiler meat in Belgium.

Acquired resistance of Escherichia coli to 3rd generation cephalosporin antimicrobials is a relevant issue in intensive broiler farming. In Belgium, about 35% of the E. coli strains isolated from live broilers are resistant to 3rd generation cephalosporins while over 60% of the broilers are found to be carrier of these 3rd generation cephalosporin resistant E. coli (CREC) after selective isolation. A model aimed at estimating the exposure of the consumer to CREC by consumption of broiler meat was elaborated. This model consists of different modules that simulate the farm to fork chain starting from primary production, over slaughter, processing and distribution to storage, preparation and consumption of broiler meat. Input data were obtained from the Belgian Food Safety agencies' annual monitoring plan and results from dedicated research programs or surveys. The outcome of the model using the available baseline data estimates that the probability of exposure to 1000 colony forming units (cfu) of CREC or more during consumption of a meal containing chicken meat is ca. 1.5%, the majority of exposure being caused by cross contamination in the kitchen. The proportion of CREC (within the total number of E. coli) at primary production and the overall contamination of broiler carcasses or broiler parts with E. coli are dominant factors in the consumer exposure to CREC. The risk of this exposure for human health cannot be estimated at this stage given a lack of understanding of the factors influencing the transfer of cephalosporin antimicrobial resistance genes from these E. coli to the human intestinal bacteria and data on the further consequences of the presence of CREC on human health.