Campylobacter contamination of broiler caeca and carcasses at the slaughterhouse and correlation with Salmonella contamination

In order to estimate the prevalence of Campylobacter spp. and Salmonella spp. on broiler chicken carcasses and the prevalence of Campylobacter spp. in caeca, 58 French slaughterhouses were investigated in 2008. Enumeration of Campylobacter spp. was also performed in order to study the relation between caeca and carcass contamination. A pool of 10 caeca and one carcass were collected from 425 different batches over a 12-month period in 2008. Salmonella was isolated on 32 carcasses leading to a prevalence of 7.5% ([5.0-10.0]_95%CI). The prevalence of Campylobacter was 77.2% ([73.2-81.2]_95%CI) in caeca and 87.5% ([84.4-90.7]_95%CI) on carcasses. No significant correlation was found between Campylobacter and Salmonella. Positive values of Campylobacter were normally distributed and the average level was 8.05 log₁₀ cfu/g ([7.94-8.16]_95%CI) in caeca and 2.39 cfu/g ([2.30-2.48]_95%CI) on carcasses. A positive correlation (r = 0.59) was found between the mean of Campylobacter in caeca and on carcasses (p < 0.001). Thus, carcasses from batches with Campylobacter-positive caeca had significantly (p < 0.001) higher numbers of Campylobacter per gram than batches with negative caeca. These results show that Campylobacter can be present in both matrices and reduction in caeca could be a possible way to reduce the amount of bacteria on carcasses. Of the 2504 identifications performed, 3 species of Campylobacter (Campylobacter jejuni, Campylobacter coli and Campylobacter lari) were identified. The main species recovered were C. jejuni and C. coli, which were isolated in 55.3% and 44.5% of positive samples, respectively. These two species were equally represented in caeca but C. jejuni was the most frequently isolated on carcasses with 57.1% and 42.5% of positive carcasses for C. jejuni and C. coli, respectively. This study underlines that target a reduction of Campylobacter on final products requires a decrease of contamination in caeca.     

Factors associated with carcass contamination by Campylobacter at slaughterhouse in cecal-carrier broilers

A study was conducted in 2009 to identify risk factors of Campylobacter spp. transmission from the digestive tract to the carcasses of standard broilers (slaughter age: 37 day, carcass weight: 1.3 kg on average). Counts of Campylobacter were performed on pools of 10 ceca and 10 neck-skins from 108 Campylobacter ceca-positive batches in three slaughterhouses. Technical and health data also was collected on the broilers: age, size, carcass weight (mean and standard deviation), condemnation rate, mortality rate and nature of treatment during the rearing period.Cecal counts varied from 4.8 to 10.2 log₁₀ cfu/g. In seventeen batches (15.7%), the skin count was below the detection limit. In the 91 batches with positive neck-skin test results, the counts varied from 2.0 to 5.2 log₁₀ cfu/g. Standard deviation of carcass weight, condemnation rate, slaughter rate and cecal count were significantly lower and growth rate higher in the 17 batches where neck-skin results were not detected positive. Multivariate analysis showed that batches with higher standard deviation of carcass weight were 5 to 9 fold more at risk of having detectable carcass contamination. Among the 91 positive neck-skin batches, only slaughter rate and cecal counts were found to have a significant but limited effect on the level of neck-skin contamination. As far as body weight homogeneity may be affected by disease, better health control can contribute to a reduction of the contamination of the broiler carcasses in Campylobacter carrier batches.     

Campylobacter contamination in broiler carcasses and correlation with slaughterhouses operational hygiene inspection

This study investigates factors associated with Campylobacter contamination of broiler carcasses, using survey data collected from nine Belgian slaughterhouses in 2008 in accordance with a European Union baseline study. Campylobacter were detected in 51.9% (202/389) (95% confidence interval, 46.8%-56.9%) of broiler carcasses. Campylobacter concentration was <10 CFU/g in 49.6% of carcasses, while 20.6% were contaminated with ≥1000 CFU/g. The mean Campylobacter concentration, as calculated by maximum likelihood estimation for left-censored data, was 1.8 log₁₀ CFU/g, with a standard deviation of 1.9 log₁₀ CFU/g. There was statistically significant variation among slaughterhouses in prevalence and concentrations of Campylobacter in their sampled carcasses. Campylobacter prevalence (but not concentrations) was positively associated with increase in broilers age. Both Campylobacter prevalence and concentration were significantly higher in carcasses sampled during June and September (but not in July and August) than carcasses sampled in January. We also investigated the correlation (Spearman’s rank correlation test) between the scores of official control inspections and Campylobacter prevalence for eight out of the nine slaughterhouses. The control inspections were routinely performed by the Belgian Federal Agency for the Safety of the Food Chain, and the concluded inspection scores were used as a general numerical indicator for the status of operational hygiene and quality of management in the slaughterhouses. Ranking of slaughterhouses based on their inspection scores was statistically correlated (Spearman’s correlation coefficient = 0.857) with their ranking based on prevalence of Campylobacter. In the present study we demonstrate how the outcomes from a routine baseline survey could be coupled with other readily available data from national control authorities in order to enable a better insight over Campylobacter contamination status in broiler slaughterhouses. Findings from this work call for subsequent in-depth investigations on technical and hygiene management factors that could impact Campylobacter contamination across broiler slaughterhouses.     

In vitro antimicrobial activity of essential oil from endemic Origanum minutiflorum on ciprofloxacin-resistant Campylobacter spp.

This study evaluated the antimicrobial activities of an essential oil of Origanum minutiflorum (O. Schwarz and P.H. Davis) against ciprofloxacin-resistant Campylobacter spp., by broth microdilution and agar well-diffusion methods. Moreover, O. minutiflorum oil was analyzed by gas chromatography/mass spectrometry (GC/MS). Twenty-nine components were identified, representing 98.7 of the oil. The oil yield from the plants was 4.0–4.4% v/w. The major components of O. minutiflorum oil were carvacrol (73.9%) and p-cymene (7.20%). The oil has lower contents of carvacrol methyl ether (0.05%), heptadecanol (0.06%) and carvacryl acetate (0.06%). Twenty-one Campylobacter spp. (12 C. jejuni, 5 C. lari and 4 C. coli) strains using in this study were selected among 300 isolates according to their resistance to ciprofloxacin. The minimum inhibitory concentration (MIC) values for bacterial strains, which were sensitive to the essential oil of O. minutiflorum, were in the range of 7.8–800 μg/ml. The essential oil obtained showed strong antimicrobial activity against all of the tested ciprofloxacin-resistance Campylobacter spp. These results suggest that the essential of O. minutiflorum may be used as a natural preservative in food against food-born disease, such as Campylobacteriosis.     

Salmonella on feces,hides and carcasses in beef slaughter facilities in Venezuela

This study determined Salmonella prevalence at different stages during the slaughtering in three beef slaughter plants (A, B and C) located in the western region of Venezuela (Zulia and Lara states). Each facility was visited three times at monthly intervals, from the months October through December of 2006. Samples were collected from hides (n = 80), fecal grabs (n = 80) and carcasses (n = 80) at the phases of pre-evisceration, after-evisceration and pre-cooler at three sampling sites on the animals (rump, flank and brisket). Salmonella prevalence was higher on hides (36.3%) than on feces (13.8%) (P < 0.05). Differences among slaughter plants for overall Salmonella prevalence were observed (P = 0.001; A: 3.5%, B: 11.1%, C: 4.4%). From the isolated strains, Salmonella enterica subspecies enterica ser. Saintpaul, Salmonella ser. Javiana and Salmonella ser. Weltevreden were identified. Cattle feces and hides might be considered as important sources of Salmonella for carcass contamination at different slaughter stages. The presence of potentially pathogenic Salmonella serotypes at the slaughtering stages is an evidence of the circulation of this pathogen in the food environment; its presence could increase consumers' risks of infection if proper food handling and preparation techniques are not followed. These data should serve as a baseline for future comparisons in Salmonella prevalence on beef carcasses to be used by the government and industry in order to establish preventive measures and to better address the risks of Salmonella contamination.     

The biofilm forming potential of bacterial species in the genus Campylobacter

The biofilm forming abilities of 16 strains representative of 14 of the 16 species comprising the genus Campylobacter were determined on glass, stainless steel, and polystyrene plastic. The formation of biofilms has been suggested as a means by which Campylobacter is able to persist within an inhospitable environment. Of the eight microaerophilic Campylobacter species, including two strains each of Campylobacter jejuni and Campylobacter fetus, only C. jejuni strain 81–176 reliably produced a visible biofilm on multiple surfaces. Alternately, all six strains of the anaerobic Campylobacter species reliably produced visible biofilms on multiple surfaces. Electron micrographs of the individual biofilms showed relatively homogeneous biofilms produced by the anaerobic strains, while the microaerophilic C. jejuni strain 81–176 produced a biofilm containing similar quantities of both the spiral and coccoid forms. This survey suggests a difference in the biofilm forming potentials and the morphologies of the bacteria comprising the biofilms between anaerobic and microaerophilic species of Campylobacter. Additionally, differences observed in the biofilm forming ability of two strains of C. jejuni suggest the need for a further investigation of the biofilm forming potential of this species using a larger number of strains.     

PorA specific primers for the identification of Campylobacter species in food and clinical samples

Campylobacteriosis is a public health problem with considerable socio-economic impact. As the European Food Safety Authority has emphasized the importance of a surveillance programme for campylobacteriosis, the aim of the present study was the optimization of a specific and sensitive PCR protocol able to detect Campylobacter species responsible for gastrointestinal infections. Raw poultry meat samples were analysed for the presence of Campylobacter sp., by plating onto mCCD (Modified Charcoal-Cefoperazone-Deoxycholate) Agar and Campylobacter Selective Preston Agar and using four sets of species-specific primers for Campylobacter jejuni, Campylobacter coli, Campylobacter upsaliensis and Campylobacter lari designed to bridge the porA gene. The resulting primers demonstrated a sensitivity of 0.01 ng/μl for the C. coli-specific, C. lari-specific, and C. upsaliensis-specific primer sets and 0.5 ng/μl for the C. jejuni-specific primer sets using DNA from pure cultures. Non-specific amplification of non-target DNA was not observed indicating excellent specificity. The primers were useful for the analyses of poultry meat samples both for direct plating onto mCCDA, and for DNA extracted directly from the cells grown for 48 h in Preston enrichment broth. The sets of primers were also useful when used for species identification of human isolates.     

Molecular characterization and antibiotic resistance profiling of Campylobacter isolated from cattle in Polish slaughterhouses

A total of 812 samples from bovine hides and the corresponding carcasses collected at the slaughterhouse level in the eastern part of Poland were examined for the presence of Campylobacter jejuni and Campylobacter coli. Recovered isolates were confirmed using species-specific PCR, characterized by the presence of 11 putative virulence genes and antimicrobial susceptibility was determined using a microbroth dilution method. Furthermore, the genotypic relatedness of the isolates was determined by PFGE profiling and virulence pattern cluster analysis. The prevalence of Campylobacter was 25.6% and 2.7% in bovine hide and carcass samples, respectively. The presence of virulence markers varied between C. jejuni and C. coli species however, the majority of strains possessed the cadF, flhA, flaA genes, irrespective of the bacterial species and origin. The lower number of the strains was positive for the invasive associated markers – virB11 and wlaN. Antibiotic profiling showed that campylobacters were most frequently resistant to quinolones and fluoroquinolones (nalidixic acid and ciprofloxacin, 38.3% of each, respectively) followed by streptomycin (24.3%) and tetracycline (20.9%). Resistance to erythromycin and gentamicin was demonstrated in 4.3% and 2.6% of strains, respectively. Comparisons of the PFGE and virulence marker profiles of the isolates reflected the high genetic diversity of Campylobacter tested. Moreover, a poor correlation between the PFGE type, pathogenic gene marker and antimicrobial resistance patterns was observed.     

Incidence and antibiotic resistance of Salmonella spp. on raw chicken carcasses

The current study was carried out to detect Salmonella spp. contamination on chicken carcasses and to determine the antibiotic susceptibility profiles and serotype distribution of the isolates. A total of 200 packaged fresh raw chicken samples sold at retail in different markets in central Anatolia were analysed between April 2005 and March 2006. Salmonella spp. was detected in 34% (68/200) of samples using cultural technique and were confirmed by PCR. Ten Salmonella serovars were identified; predominant ones included Typhimurium, Infantis and Heidelberg. All of the Salmonella spp. isolates tested, exhibited resistance to one or more antimicrobial agents used. Resistance to penicillin, oxacillin, clindamycin, vancomycin, erythromycin and ampicillin were evident 100%, 97%, 97%, 92.6%, 89.7% and 85.2%, respectively. Also resistance to tetracycline (67.6%), streptomycin (61.7%), neomycin (55.8%) and cephalothin (52.9%) was observed but a small percentage of the isolates demonstrated resistance to gentamicin (14.7%), chloramphenicol (10.2%), cefotaxime (2.9%) and amikacin (2.9%). As a result, high prevalence of Salmonella spp. and the relatively high resistance among the bacteria tested could pose public health and therapeutic problems in consumers as potential vehicle of resistant Salmonella foodborne infections. To avoid Salmonella contamination, hygienic rules of slaughter and poultry meat processing must be rigorously observed and antibiotic use must be controlled by governmental agencies to prevent increased resistance of antibiotics.     

The effects of Campylobacter numbers in caeca on the contamination of broiler carcasses with Campylobacter

For the presence and number of Campylobacter, 18 broiler flocks were sampled over a period of 18 months. A total of 70% of the flocks were positive for Campylobacter, with higher prevalence found in summer and autumn, compared to winter and spring. Positive flocks showed contamination rates above 90%, in negative flocks this was lower, mostly below 50%. The enumeration showed a decrease in Campylobacter during processing of positive flocks. The numbers were highest in carcasses after scalding/defeathering (mean 5.9 log(10) cfu/carcass) and dropped by 0.7 log(10) cfu/carcass after chilling. A positive correlation was observed between the number of Campylobacter present in the caeca and the number of bacteria present on carcasses and cut products. When a negative flock was slaughtered after Campylobacter positive flocks, the number of positive samples was higher compared to the case when a negative flock had been slaughtered previously. C. jejuni was isolated from 73.6% of the poultry samples.     

Prevalence and risk factors for bruises in Chilean bovine carcasses

Records of cattle slaughtered at two Chilean slaughterhouses (SLH1 and SLH2) were used to determine prevalence and risk factors for carcasses with bruises. Bruise prevalence amounted to 12.3% but differed between slaughterhouses (20.8% for SLH1 and 8.6% for SLH2 respectively). Bruise severity grade 1 (mild) was most frequently recorded. The type of the animal, source of animal, the level of fat cover and lairage time were associated with the presence of bruises. Older categories of animals and animals that pass through a market before being moved to the slaughterhouse are more prone to show bruises. The results also indicate that under the reported Chilean circumstances animals that have longer lairage times (over 12 h) have a significantly reduced risk for bruises, except for oxen. Presence of bruises is also significantly associated with increased carcass pH values.     

Diversity of culturable microorganisms and occurrence of Listeria monocytogenes and Salmonella spp. in Tuber aestivum and Tuber melanosporum ascocarps

The aim of this study was to investigate the total mesophilic microorganisms, Pseudomonas genus, Enterobacteriaceae family, mold and yeast counts and the presence of Listeria monocytogenes and Salmonella spp on Tuber aestivum and Tuber melanosporum ascocarps. The results confirmed that the major percentage of the microorganisms, approximately 9.0 log ufc/g, were present in the peridium, the glebas of healthy truffles being practically free of microorganisms. The predominant microbial group was the Pseudomonas averaging 8.3 and 8.4 log cfu/g on T. aestivum and T. melanosporum whole ascocarps, respectively. The Enterobacteriaceae also achieved high populations, especially in T. aestivum truffles, with 6.3 log cfu/g. Molds and yeasts never exceeded 5.0 log cfu/g. The characterization of the isolates revealed that the fluorescens pseudomonads were the most prevalent. Raoultella terrigena and Enterobacter intermedius were the dominant Enterobacteriaceae. The identification of the yeast isolates revealed five species: Debaryomyces hansenii, Issatchenkia scutulata, Rhodotorula aurantiaca, Saccharomyces dairensis and Trichosporon beigelii subspecies A and B. The mold genera detected in both species of truffles were Aspergillus, Cladosporium, Penicillium and Fusarium, Trichoderma being present only in T. aestivum. L. monocytogenes was found in 10% of the samples of T. aestivum analysed but Salmonella spp. was not detected. Knowledge of the microbial population in terms of possible food borne and pathogen microorganisms is very useful for establishing successful disinfection and storage methods to prolong the shelf-life of ascocarps of T. aestivum and T. melanosporum.     

Occurrence of Shiga toxin-producing E. coli (STEC) on carcasses and retail beef cuts in the marketing chain of beef in Argentina

Argentina has the highest incidence of HUS in the world. HUS is produced by STEC O157 and non-O157. Cattle's faeces and hides are sources of STEC contamination of carcasses during slaughter. We investigated the presence of STEC in carcasses and cuts of meat in the marketing chain in an agricultural city located in Buenos Aires Province (Argentina). In this study, the detection of the stx gene was used as an indicator of carriage of meat with STEC. In carcasses, we detected 12.34% and 18.64% of STEC at the slaughter and sanitary control cabin (place where carcasses arrive from slaughters located outside the city), respectively. These percentages increased at butcheries (24.52%). The 25% of retail beef cuts were STEC-positive with significant differences among the different cuts of meat (chuck: 12.12%, rump roast: 12.12% and minced beef: 40.74%). The stx2 gene was the predominant gene detected in all samples at different levels of the commercialization meat chain.     

Assessing the cross contamination and transfer rates of Salmonella enterica from chicken to lettuce under different food-handling scenarios

Cross contamination of foodborne pathogens from raw meats to ready-to-eat foods has caused a number of foodborne outbreaks. The cross contamination and transfer rates of Salmonella enterica from chicken to lettuce under various food-handling scenarios were determined. The following scenarios were tested: in scenario 1, cutting board and knife used to cut chicken (10⁶ CFU/g) were also used for cutting lettuce, without washing; in scenario 2, cutting board and knife were washed with water separately after cutting chicken, and subsequently used for cutting lettuce; and in scenario 3, cutting board and knife were thoroughly washed with soap and hot water after cutting chicken, and before cutting lettuce. In each scenario, cutting board, knife, chicken and lettuce were sampled for population of S. enterica. For scenario 1, both before and after cutting lettuce, the cutting board and knife each had about 2 logs CFU/cm² of S. enterica, respectively. The cut lettuce had about 3 logs CFU/g of S. enterica. In scenario 2, fewer organisms (0.5–2.4 logs CFU/g or cm²) were transferred. The transfer rates in both scenarios ranged from 0.02 to 75%. However, in scenario 3, <1 log CFU/g or cm² organisms were detected on lettuce, cutting board or knife, after washing and cutting lettuce. This shows that the FDA recommended practice for cleaning cutting boards is effective in removing S. enterica and preventing cross contamination.     

Evaluation of ISO 10272:2006 standard versus alternative enrichment and plating combinations for enumeration and detection of Campylobacter in chicken meat

In the present study, we evaluate the recommended ISO 10272:2006 versus alternative procedures for Campylobacter enumeration and enrichment in naturally contaminated chicken meat samples (n = 49). Three enrichment media were evaluated; Bolton broth, Preston broth and CampyFood broth^® (bioMérieux SA, Marcy l’Etoile, France). In addition, three selective plating agars were compared; modified charcoal cefoperazone deoxycholate agar (mCCDA), CampyFood agar^® (CFA; bioMérieux SA) and Brilliance CampyCount agar^® (BCC; Oxoid, Basingstoke, England). Direct plating on CFA provided the highest number of Campylobacter positive samples (17/49); however this was not statistically different (P > 0.05) from numbers of positive samples recovered by direct plating on mCCDA (15/49) or BCC agars (14/49). Also, there was no significant difference between Campylobacter counts on the three compared media (P > 0.05). The coloured colonies of Campylobacter on CFA and BCC were easier to record and count than those on mCCDA. Enrichment of chicken meat samples in Bolton broth for 48 h and subsequent plating on CFA provided significantly higher (P < 0.05) Campylobacter detection compared to the other broth-agar combinations. Enrichment in Preston broth for 24 h followed by plating on mCCDA gave a higher number of positive samples (20/49) than 48 h enrichment in Bolton broth and plating on mCCDA (15/49). Enrichment in Bolton broth for 48 h followed by plating on CFA recovered 35% of samples below the limit for quantifications (<10 CFU/g, n = 34), as identified by direct plating on mCCDA. Compared to the current ISO method, some alternative combinations of enrichment and agar media could provide significantly better detection and enumeration of Campylobacter in chicken meat.     

Consumption of raw vegetables and fruits: a risk factor for Campylobacter infections

The purpose of this study was to determine the prevalence of Campylobacter in fresh vegetables and fruits at retail level in the Netherlands, and to estimate its implications on the importance of vegetables and fruits as risk factor for campylobacteriosis.Thirteen of the 5640 vegetable and fruit samples were Campylobacter positive, resulting in a prevalence of 0.23% (95% confidence interval (Cl): 0.12-0.39%). The prevalence of packaged products (0.36%, 95% Cl: 0.17-0.66) was significantly higher than of unpackaged products (0.07; 95% Cl: 0.01-0.27). No statistical differences were found between seasons.Combining the mean prevalence found in this study with data on the consumption of vegetables and fruits, an exposure of 0.0048 campylobacters ingested per person per day in the Netherlands by transmission via vegetables and fruits, was calculated. This exposure, as input in a Beta-Poisson dose-response model, resulted in an estimated number of 5.3 × 10⁵ cases of infection with Campylobacter per year for the whole Dutch population. This constitutes the consumption of raw vegetables and fruits, especially when packaged, to be a risk factor for Campylobacter infections.     

Prevalence and contamination patterns of Listeria monocytogenes in catfish processing environment and fresh fillets

Catfish skins, intestines, fresh fillets, processing surfaces at different production stages, chiller water and non-food contact surfaces were sampled for Listeria monocytogenes and other Listeria species. Among 315 samples, prevalence of L. monocytogenes, Listeria innocua and a group of Listeria seeligeri–Listeria welshimeri–Listeria ivanovii was 21.6, 13.0 and 29.5%, respectively. No Listeria grayi was detected in this survey. While no L. monocytogenes strains were isolated from catfish skins and intestines, the strains were found with a frequency of 76.7% in chilled fresh catfish fillets and 43.3% in unchilled fillets. L. monocytogenes and Listeria spp. were also detected in fish contact surfaces such as deheading machine, trimming board, chiller water, conveyor belts at different stages, and fillet weighing table. Among L. monocytogenes, 1/2b (47.0%), 3b (16.0%) and 4c (14%) were the predominant serotypes isolated, whereas 4b, 4e, 1/2c and 1/2a were detected at much lower frequencies. Genotype analyses of L. monocytogenes isolates using serotyping, pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus (ERIC)-PCR revealed that chiller water represented an important contamination source of L. monocytogenes in the chilled catfish fillets of two processing facilities, whereas fillet weighing table significantly contributed to the catfish fillet contamination of the third facility. This study suggests that L. monocytogenes contamination in the processed catfish fillets originates from the processing environment, rather than directly from catfish. Results from this study can aid the catfish industry to develop a plant-specific proper cleaning and sanitation procedure for equipment and the processing environment designed to specifically target L. monocytogenes contamination.     

Preliminary virulence genotyping and phylogeny of Escherichia coli from the gut of pigs at slaughtering stage in Brazil

In the current study we screened Escherichia coli from intestine of pigs slaughtered in Mato Grosso, Brazil, for virulence-markers related to human disease. Furthermore, we employed for the first time a phylogenetic assay to explore the association between phylogeny and virulence genotype in E. coli from finished swine. A low prevalence (7.8%) of E. coli harbouring virulence genes was observed. Among the positive isolates, 3.3% could be classified as atypical EPEC, 2.2% as STEC and 2.2% as CDT harbouring E. coli. Virulence genes were not found to co-occur in a strain. Phylogenetic determination of isolates revealed a low prevalence of E. coli lineages related to disease. Therefore, preliminary sampling of 74 pigs indicated that slaughter swine may not be major reservoirs of E. coli capable of causing human disease. In light of the significant association between phylogeny and virulence genotype, we also underscored the phylogenetic grouping of strains as a valuable tool for E. coli surveillance programmes in slaughterhouses.     

Presence of Clostridium difficile in pigs and cattle intestinal contents and carcass contamination at the slaughterhouse in Belgium

The objective of this study was to evaluate the presence of Clostridium difficile in intestinal and carcass samples collected from pigs and cattle at a single slaughterhouse. C. difficile was isolated in 1% and 9.9% of the pig and cattle intestinal contents and in 7.9% and 7% of cattle and pig carcass samples respectively. A total of 19 different PCR-ribotypes were identified, among them types 078 and 014. Seven of 19 ribotypes correlated with the PCR-ribotypes involved in human C. difficile infections in Belgium. This study confirms that animals are carriers of C. difficile at slaughter and ribotypes are identical than those in humans, and that carcass contamination occurs inside the slaughterhouse.