Decontamination of knives used in a slaughterhouse by a commercial non-thermal UV-C treatment.

To assess the antimicrobial effect of a commercial UV-C system, knives inoculated with Escherichia coli and Staphylococcus aureus as well as naturally contaminated and collected from the wet and clean area of a slaughterhouse knives were examined. For inoculated knives, UVC treatment for 30 s reduced mean E. coli counts by 5.1 log CFU cm-2 and mean S. aureus counts by 4.5 log CFU cm-2. The presence of blood lowered mean reductions to 3.4 log CFU cm-2 for E. coli and to 2.5 log CFU cm-2 for S. aureus. The presence of fat had a greater negative impact on the efficacy of the UV-C treatment resulting in mean reductions <1.8 log CFU cm-2. For naturally contaminated knives from a slaughterhouse, total viable counts (TVC) before UV-C treatment varied considerably (wet area: 2.0-6.0 log CFU cm-2, clean area: 1.0-3.0 log CFU cm-2). UV-C treatment for 30s reduced mean TVC by 0.8 log CFU cm-2 (wet area) and 0.6 log CFU cm-2 (clean area), but the effect varied greatly between individual knives. Thus, under commercial conditions, the antibacterial effect of UV-C for the decontamination of knives is affected by the presence of additional contaminations like blood or fat. The adequate cleaning of the knives prior to UV-C decontamination is therefore of central importance.

Microbiological quality and presence of foodborne pathogens in raw milk cheeses and raw meat products marketed at farm level in Switzerland.

This study investigated the microbiological quality and presence of bacterial foodborne pathogens in 51 raw milk cheeses (mainly semihard and hard cheese) and 53 raw meat products (cured meat products and sausages) marketed at farm level. With regard to Enterobacteriaceae, Escherichia (E.) coli, and coagulase-positive staphylococci (CPS), the examined products were generally of a good microbiological quality. Enterobacteriaceae were found in seven cheeses (1.0×102 - 8.8×104 CFU/g) and one sausage (2.0×102 CFU/g). Three of these cheeses were also positive for E. coli. CPS results were comparable for cheeses (5.9%; 1.0-6.0×102 CFU/g) and meat products (3.8%; 1.0-2.0×102 CFU/g). On the other hand, such raw products may harbor potential health hazards as Listeria (L.) monocytogenes, Shiga toxin-producing E. coli (STEC), and staphylococcal enterotoxin (SE)-producing Staphylococcus (S.) aureus. L. monocytogenes were found in one sausage and the isolate belonged to the serotype 1/2c. The two STEC isolates harbored stx1a (cheese) or stx2e (sausage), but both lacked eae and did not belong to the top five-serogroups. Of the five S. aureus isolates, the three cheese isolates belonged to the clonal complex (CC) 8, CC22, and CC705, the two sausage isolates belonged to CC7, and all isolates harbored genes for SEs. Thus, to avoid contaminations and to prevent foodborne pathogens from entering the food chain, strict compliance with good hygiene practices during milk and cheese production or meat production is of central importance.

Effect of a commercial steam-vacuuming treatment implemented after slaughtering for the decontamination of cattle carcasses.

To assess the antimicrobial effect of a commercial steam-vacuuming system newly implemented after slaughtering, 105 cattle carcasses were examined for total viable counts (TVC) at four different areas. Before steam vacuuming, mean TVC of the excision samples were comparable at the perineal area and brisket (3.0-3.1 log CFU cm-2) or the hind leg and shoulder (2.6-2.7 log CFU cm-2). Steam vacuuming reduced mean TVC by 0.9, 0.7, 0.6, and 0.4 log CFU cm-2 at the perineal area, hind leg, shoulder, and brisket, respectively. With regard to the distribution of counts, steam vacuuming increased the proportion of TVC results <3.0 log CFU cm-2 from 74.8% (62.9-87.6% at carcass areas) to 86.7% (71.4-97.1% at carcass areas). Thus, steam vacuuming after slaughtering might be useful for the reduction of contamination in designated carcass areas, but the effect must not be overestimated and decontamination treatments always must be seen part of an integral food safety system.

Characteristics of Shiga Toxin-Producing Escherichia coli O157 in Slaughtered Reindeer from Northern Finland.

Fecal samples collected from 470 slaughtered reindeer 6 to 7 months of age were screened by real-time PCR (after enrichment) for Shiga toxin genes (stx) and then for Escherichia coli serogroup O157. Shiga toxin genes were found frequently (>30% of samples), and serogroup O157 was detected in 20% of the stx-positive samples. From these samples, a total of 25 E. coli O157:H- isolates (nonmotile but PCR positive for fliCH7) were obtained. Twenty-four of these E. coli O157:H- isolates did not ferment sorbitol and originated from one geographic area. These 24 isolates belonged to the multilocus sequence type 11, typical for Shiga toxin-producing E. coli (STEC) O157:H7 and O157:H-, and harbored genes stx1a, stx2c, eae, and hlyA; the stx2c subtype has been associated with high virulence. In contrast, one E. coli O157:H- isolate (multilocus sequence type 11) did ferment sorbitol, lacked Shiga toxin genes, but was positive for eae, hlyA, and sfpA. This isolate closely resembled an STEC that has lost its Shiga toxin genes. Additional examination revealed that reindeer can be colonized by various other STEC isolates; 21 non-O157 STEC isolates belonged to four multilocus sequence types, harbored stx1a (8 isolates) or stx2b (13 isolates), and in the stx2b-positive isolates the recently described new allelic variants (subAB2-2 and subAB2-3) for subtilase cytotoxin were identified. Hence, slaughtered semidomesticated Finnish reindeer might constitute a little known reservoir for STEC O157:H7/H- and other serogroups, and the risk of direct or indirect transmission of these pathogens from reindeer to humans and domestic livestock must not be overlooked.

Presence of foodborne pathogens, extended-spectrum ß-lactamase -producing Enterobacteriaceae, and methicillin-resistant Staphylococcus aureus in slaughtered reindeer in northern Finland and Norway.

BACKGROUND: Various food-producing animals were recognized in recent years as healthy carriers of bacterial pathogens causing human illness. In northern Fennoscandia, the husbandry of semi-domesticated reindeer (Rangifer tarandus tarandus) is a traditional livelihood and meat is the main product. This study determined the presence of selected foodborne pathogens, methicillin-resistant Staphylococcus aureus (MRSA), and extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae in healthy semi-domesticated reindeer at slaughter in northern Finland and Norway. RESULTS: All 470 reindeer fecal samples tested negative for Salmonella spp., whereas L. monocytogenes was detected in 3%, Yersinia spp. in 10%, and Shiga toxins genes (stx1 and/or stx2) in 33% of the samples. Listeria monocytogenes isolates belonged to the serotype 1/2a (14/15) and 4b, Yersinia spp. were identified mainly as Y. kristensenii (30/46) and Y. enterocolitica (8/46), and stx2 predominated among the Shiga toxin genes (stx2 alone or in combination with stx1 was found in 25% of the samples). With regard to the frequency and distribution of stx1/stx2, striking differences were evident among the 10 different areas of origin. Hence, reindeer could constitute a reservoir for Shiga toxin-producing E. coli (STEC), but strain isolation and characterization is required for verification purposes and to assess the potential human pathogenicity of strains. On the other hand, the favorable antibiotic resistance profiles (only 5% of 95 E. coli isolates were resistant to one or more of the tested antibiotics) and the absence of MRSA and ESBL-producing Enterobacteriaceae (when applying selective methods) suggest only a limited risk of transmission to humans. CONCLUSIONS: Healthy semi-domesticated reindeer in northern Finland and Norway can be carriers of certain bacterial foodborne pathogens. Strict compliance with good hygiene practices during any step of slaughter (in particular during dehiding and evisceration) is therefore of central importance to avoid carcass contamination and to prevent foodborne pathogens from entering the food chain.

Analysis of a poultry slaughter process: Influence of process stages on the microbiological contamination of broiler carcasses.

In a large-scale Swiss poultry abattoir, a microbiological process analysis of broiler carcasses was performed. At each selected process stage (scalding, plucking, evisceration, washing, and chilling), 90 carcasses from 30 flocks were sampled and examined for Campylobacter, Salmonella, Escherichia coli, Enterobacteriaceae, and extended-spectrum b-lactamases-producing Enterobacteriaceae. With regard to Campylobacter counts on carcasses, plucking tended to slightly increase the results (on average by 0.4 log CFU/g), whereas mean counts from plucked and chilled carcasses were comparable (3.1 log CFU/g after plucking, 3.0 log CFU/g in the chiller). The Campylobacter results of chilled carcasses are thereby likely to comply with the newly defined requirements of the European Union (process hygiene criterion for Campylobacter). With regard to Escherichia coli and Enterobacteriaceae counts on carcasses, plucking clearly reduced the results (on average by 0.8 and 0.9 log CFU/g), whereas mean counts from plucked and chilled carcasses were comparable (3.4 and 3.5 log CFU/g after plucking, 3.4 log CFU/g in the chiller). In contrast, Salmonella spp. were not detected on broiler carcasses and extended-spectrum b-lactamases- producing Enterobacteriaceae only rarely (1.8%). Such abattoir-specific data are of central importance for assessment of slaughter process performance and if necessary for the implementation of effective measures in the slaughter process.

Microbiological contamination of cattle carcasses at different stages of slaughter in two abattoirs.

Cattle carcasses from two abattoirs were examined at selected stages of slaughter (skinning, evisceration, trimming, washing, blast chilling) for aerobic colony counts (ACC) and Enterobacteriaceae. At each stage and abattoir, 50 carcasses were sampled by swabbing at the neck, brisket, flank and rump. After skinning, average ACC on carcasses was 1.5logCFUcm(-2) and Enterobacteriaceae frequencies at sites were ≤6%. From skinned to washed carcasses, certain abattoir- and site-specific changes occurred. Blasting clearly reduced ACC and Enterobacteriaceae results on carcasses from abattoir B, but reductions were limited or lacking in abattoir A. In addition, 100 hides and corresponding chilled carcasses were examined. On hides, average ACC was 5.6logCFUcm(-2) and Enterobacteriaceae frequencies at sites ranged from 74 to 96%. Average carcass-hide ratios of the two abattoirs were comparable for ACC (0.0182-0.0202%) but differed for Enterobacteriaceae counts (abattoir A: 0.4627%; abattoir B: 0.0941%). Such ratios allow comparing process performance between abattoirs in the daily practice.

Microarray-based characterization of Staphylococcus aureus isolates obtained from chicken carcasses.

A total of 34 Staphylococcus aureus isolates from flock-wise pooled chicken neck skin samples collected at two abattoirs during slaughter were characterized with DNA microarray analysis and spa typing. The 20 isolates from abattoir A all belonged to clonal complex (CC) 12 and spa type t160. Of the 14 isolates from abattoir B, 7 belonged to CC5-t3478, 5 to CC12-t160, 1 to CC45-t040, and 1 to CC101-t056. Of the various resistance-associated genes tested, only blaZ/R/I (6 isolates of CC12 and CC101 from abattoir B), sdrM (n = 34), fosB (n = 33), and qacC (n = 22) were detected. None of the isolates harbored genes conferring methicillin resistance. In terms of genes encoding enterotoxins, seb (all isolates of CC12), egc (seg, sei, selm, seln, selo, selu; all isolates of CC5 and CC45), and sea (14 isolates of CC12 and 1 isolate of CC5) were found. In addition, all isolates harbored genes for intracellular adhesion proteins (icaA/C/D) and were positive for cap5 or cap8 (capsule type 5 or 8). Comparison of DNA microarray profiles identified four categories comprising (i) all isolates of CC12, (ii) all isolates of CC5, (iii) the CC45 isolate, and (iv) the CC101 isolate. The high similarity of the isolates from abattoir A could indicate contamination of chicken carcasses with S. aureus persisting on the slaughter equipment, but further investigations are required to elucidate potential contamination routes.

Detection of the emerging Shiga toxin-producing Escherichia coli O26:H11/H- sequence type 29 (ST29) clone in human patients and healthy cattle in Switzerland.

Shiga toxin-producing Escherichia coli O26:H11/H(-) strains showing the characteristics of the emerging human-pathogenic ST29 clone (stx2a(+) only, eae(+), plasmid gene profile hlyA(+) etpD(+)) were detected from human patients and healthy cattle, indicating a possible reservoir. Sheep also appear to shed strains related to the new pathogenic clone O26:H11/H(-) (ST29, stx1a(+) only, eae(+), plasmid gene profile hlyA(+) etpD(+)).

Evaluation of seven different commercially available real-time PCR assays for detection of shiga toxin 1 and 2 gene subtypes.

Following the recent outbreak of Shiga toxin-producing Escherichia coli (STEC) O104:H4 infection in Germany, the demand for fast detection of STEC has again increased. Various real-time PCR-based methods enabling detection of Shiga toxin genes (stx) have been developed and can be used for applications in food microbiology. The present study was conducted to evaluate the reliability of seven commercially available real-time PCR systems for detection of stx1 and stx2 subtypes. For this purpose, pure cultures of 18 STEC strains harboring all known stx1 and/or stx2 subtypes were tested. Only one of the seven real-time PCR systems detected all known stx1 and stx2 subtypes. Six systems failed to detect the stx2f subtype. One system missed stx2 subtypes reported in association with severe human disease. Because the presence of certain stx genes (subtypes) is considered an important indicator of STEC virulence, systems differentiating between the stx1 and stx2 gene groups provide added value. Reliable and fast detection of stx genes is of major importance for both diagnostic laboratories and the food industry.

ESBL-producing uropathogenic Escherichia coli isolated from dogs and cats in Switzerland.

Extended-spectrum ß-lactamase (ESBL)-producing Escherichia (E.) coli have emerged in human and veterinary medicine. The aim of this study was to investigate the presence of ESBL-producers among uropathogenic E. coli isolated from dogs and cats and to characterize detected ESBL-producing isolates by antibiotic susceptibility testing, identification of ESBL genes, multi-locus sequence typing (MLST), detection of putative virulence genes, and analysis of E. coli phylogroups. Among the 107 E. coli isolates derived from urinary samples (59 from dogs, 40 from cats), eight isolates from four different animals (two dogs, two cats) were found to be ESBL-producers. These isolates were of ST533/CTX-M-15/TEM/phylogroup B1 (four strains from one dog), ST410/CTX-M-15/TEM/phylogroup A (three strains, one from a dog and two from a cat), and ST648/CTX-M-15/phylogroup D (one strain from a cat). In terms of putative virulence factors, all isolates harbored lpfA, sat, and tsh, whereas iss was only detected in strains of ST533. Thus, ESBL-producers were detected among uropathogenic E. coli from Swiss companion animals and the eight CTX-M-15-producing isolates belonged to three sequence types (ST410, ST533, ST648) and three E. coli phylogroups (A, B1, D). For the first time, E. coli of ST533 carrying bla(CTX-M-15) were thereby detected in a dog.

Shedding of foodborne pathogens and microbial carcass contamination of hunted wild ruminants.

To assess the shedding of selected bacterial foodborne pathogens, fecal samples from 239 hunted wild red deer, roe deer, chamois, and ibex were examined. All samples tested negative for Salmonella spp. and L. monocytogenes, but other Listeria species were occasionally found. Of the 239 fecal samples, 32.6% tested positive for stx (Shiga toxins), 6.7% for eae (intimin) and 13.8% for both stx and eae genes. Among the 56 isolated Shiga toxin-producing Escherichia coli (STEC) strains, 44.6% harbored genes for the Stx2 group, 30.4% for the Stx1 group, and 21.4% for both Stx1 and Stx2. Only two of these strains harbored eae. Hence, wild ruminants constitute a reservoir for STEC, but further characterization data of the isolated strains are required to assess their actual human pathogenicity. In addition, 328 carcasses from hunted wild red deer, roe deer, and chamois were examined for total viable counts (TVC) and Enterobacteriaceae by swabbing. For the examined animal species, average TVC (4.0-4.2 log CFU cm(-2)) and average Enterobacteriaceae counts/detection rates (2.3-2.6 log CFU cm(-2); 87.5-90%) were at comparable levels. On the other hand, the microbial status of carcasses differed between certain abattoirs by several orders of magnitude. Strict compliance with good hunting and hygiene practices during any step from shooting, through evisceration in the field, to dehiding, cooling, and processing is therefore of central importance to avoid contaminations and to prevent foodborne pathogens carried by the animals from entering the food chain.

Prevalence and characteristics of methicillin-resistant coagulase-negative staphylococci from livestock, chicken carcasses, bulk tank milk, minced meat, and contact persons.

BACKGROUND: Methicillin-resistant coagulase-negative staphylococci (MR-CNS) are of increasing importance to animal and public health. In veterinary medicine and along the meat and milk production line, only limited data were so far available on MR-CNS characteristics. The aim of the present study was to evaluate the prevalence of MR-CNS, to identify the detected staphylococci to species level, and to assess the antibiotic resistance profiles of isolated MR-CNS strains. RESULTS: After two-step enrichment and growth on chromogenic agar, MR-CNS were detected in 48.2% of samples from livestock and chicken carcasses, 46.4% of samples from bulk tank milk and minced meat, and 49.3% of human samples. Using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), 414 selected MR-CNS strains belonged to seven different species (S. sciuri, 32.6%; S. fleurettii, 25.1%; S. haemolyticus, 17.4%; S. epidermidis, 14.5%, S. lentus, 9.2%; S. warneri, 0.7%; S. cohnii, 0.5%). S. sciuri and S. fleurettii thereby predominated in livestock, BTM and minced meat samples, whereas S. epidermidis and S. haemolyticus predominated in human samples. In addition to beta-lactam resistance, 33-49% of all 414 strains were resistant to certain non-beta-lactam antibiotics (ciproflaxacin, clindamycin, erythromycin, tetracycline). CONCLUSIONS: A high prevalence of MR-CNS was found in livestock production. This is of concern in view of potential spread of mecA to S. aureus (MRSA). Multiresistant CNS strains might become an emerging problem for veterinary medicine. For species identification of MR-CNS isolated from different origins, MALDI-TOF MS proved to be a fast and reliable tool and is suitable for screening of large sample amounts.

Characterization of attaching and effacing Escherichia coli (AEEC) isolated from pigs and sheep.

BACKGROUND: Attaching and effacing Escherichia coli (AEEC) are characterized by their ability to cause attaching-and-effacing (A/E) lesions in the gut mucosa of human and animal hosts leading to diarrhoea. The genetic determinants for the production of A/E lesions are located on the locus of enterocyte effacement (LEE), a pathogenicity island that also contains the genes encoding intimin (eae). This study reports data on the occurrence of eae positive E. coli carried by healthy pigs and sheep at the point of slaughter, and on serotypes, intimin variants, and further virulence factors of isolated AEEC strains. RESULTS: Faecal samples from 198 finished pigs and 279 sheep were examined at slaughter. The proportion of eae positive samples was 89% for pigs and 55% for sheep. By colony dot-blot hybridization, AEEC were isolated from 50 and 53 randomly selected porcine and ovine samples and further characterized. Strains of the serotypes O2:H40, O3:H8 and O26:H11 were found in both pigs and sheep. In pigs O2:H40, O2:H49, O108:H9, O145:H28 and in sheep O2:H40, O26:H11, O70:H40, O146:H21 were the most prevalent serotypes among typable strains. Eleven different intimin types were detected, whereas gamma2/theta was the most frequent, followed by beta1, epsilon and gamma1. All but two ovine strains tested negative for the genes encoding Shiga toxins. All strains tested negative for the bfpA gene and the EAF plasmid. EAST1 (astA) was present in 18 of the isolated strains. CONCLUSION: Our data show that pigs and sheep are a source of serologically and genetically diverse intimin-harbouring E. coli strains. Most of the strains show characteristics of atypical enteropathogenic E. coli. Nevertheless, there are stx-negative AEEC strains belonging to serotypes and intimin types that are associated with classical enterohaemorrhagic E. coli strains (O26:H11, beta1; O145:H28, gamma1).

Occurrence and genotypes of Campylobacter in broiler flocks, other farm animals, and the environment during several rearing periods on selected poultry farms.

On 15 Swiss poultry farms, broiler flocks, other farm animals, and the environment were examined during consecutive rearing periods to investigate the occurrence and genetic diversity of Campylobacter. Of the 5154 collected samples, 311 (6%) from 14 farms were Campylobacter positive by culture. Amongst the positive samples, 228 tested positive for Campylobacter jejuni and 92 for Campylobacter coli. Positive samples originated from broilers, the broiler houses, cattle, pigs, bantams, laying hens, a horse, and a mouse. Feed, litter, flies, and the supply air to the broiler house tested negative. By flagellin gene typing (fla-RFLP) and pulsed-field gel electrophoresis (PFGE), 917 Campylobacter isolates were genotyped. Additionally, amplified fragment length polymorphism (AFLP) analysis was performed on 15 assorted strains. On eight farms, matching genotypes were isolated from broiler flocks and other farm animals: Certain genotypes from cattle (farms H, K, L, and M), pigs (farms D and P), or laying hens (farm L) were subsequently found in the broiler flocks, whereas other genotypes initially present in the broiler flocks turned up in cattle (farms A, D, and O). These results emphasize the importance of other farm animals on poultry farms for broiler flock colonization. Indications of persistent contamination of the broiler house were evident on four farms (C, D, I, and L) where matching genotypes were detected in consecutive broiler flocks, but not concurrently in other samples. By fla-RFLP, PFGE, and confirmed by AFLP, some genotypes proofed to be identical across different farms.

Activatable Shiga toxin 2d (Stx2d) in STEC strains isolated from cattle and sheep at slaughter.

Shiga toxin producing Escherichia coli (STEC) harbouring the stx(2d-activatable) gene and expressing the mucus- and elastase-activatable phenotype have been associated with severe outcomes of human disease. However, there is limited data available on the occurrence of such strains in livestock reservoirs. In this study, we analyzed 11 STEC strains isolated from healthy cattle and sheep at slaughter that were originally detected to contain the stx(2c) allele, for the presence of the stx(2d-activatable) genotype. Ten of the eleven strains displayed the stx(2d-activatable) genotype as determine by PstI restriction fragment length polymorphism (RFLP) of 890-bp fragments of their stx genes. However, only in 6 of the 10 strains whose stx genes were sequenced, the presence of stx(2d-activatable) could be confirmed based on the predicted amino acid sequence of their StxA subunits; the remaining four strains contained Stx2c A subunit. Five of the six strains which contained stx(2d-activatable) displayed the activatable phenotype on Vero cells. Genes for adhesins such as the outer membrane protein intimin (eae), which is essential for the intimate attachment and the formation of attaching-and-effacing lesions on intestinal epithelial cells, or the STEC autoagglutinating adhesin (saa), potentially important in eae-negative STEC, were not detected. Moreover, all the strains tested negative for EHEC-hlyA encoding enterohaemorrhagic E. coli (EHEC) hemolysin. To our knowledge, this is the first study that reports the presence of STEC harbouring stx(2d-activatable) and producing the activatable Stx2d in fecal samples of sheep. Therefore both cattle and sheep are reservoirs of such strains and potential sources of human infections. This is of particular importance, because in contrast to other eae-negative STEC, strains producing Stx2d(activatable) may cause severe diseases such as bloody diarrhoea and haemolytic uremic syndrome in humans.

Serotypes, intimin variants and other virulence factors of eae positive Escherichia coli strains isolated from healthy cattle in Switzerland. Identification of a new intimin variant gene (eae-eta2).

BACKGROUND: Enteropathogenic Escherichia coli (EPEC) and Shigatoxin-producing Escherichia coli (STEC) share the ability to introduce attaching-and-effacing (A/E) lesions on intestinal cells. The genetic determinants for the production of A/E lesions are located on the locus of enterocyte effacement (LEE), a pathogenicity island that also contains the genes encoding intimin (eae). This study reports information on the occurrence of eae positive E. coli carried by healthy cattle at the point of slaughter, and on serotypes, intimin variants, and further virulence factors of isolated EPEC and STEC strains. RESULTS: Of 51 eae positive bovine E. coli strains, 59% were classified as EPEC and 41% as STEC. EPEC strains belonged to 18 O:H serotypes, six strains to typical EPEC serogroups. EPEC strains harbored a variety of intimin variants with eae-beta1 being most frequently found. Moreover, nine EPEC strains harbored astA (EAST1), seven bfpA (bundlin), and only one strain was positive for the EAF plasmid. We have identified a new intimin gene (eta2) in three bovine bfpA and astA-positive EPEC strains of serotype ONT:H45. STEC strains belonged to seven O:H serotypes with one serotype (O103:H2) accounting for 48% of the strains. The majority of bovine STEC strains (90%) belonged to five serotypes previously reported in association with hemolytic uremic syndrom (HUS), including one O157:H7 STEC strain. STEC strains harbored four intimin variants with eae-epsilon1 and eae-gamma1 being most frequently found. Moreover, the majority of STEC strains carried only stx1 genes (13 strains), and was positive for ehxA (18 strains) encoding for Enterohemolysin. Four STEC strains showed a virulence pattern characteristic of highly virulent human strains (stx2 and eae positive). CONCLUSION: Our data confirm that ruminants are an important source of serologically and genetically diverse intimin-harboring E. coli strains. Moreover, cattle have not only to be considered as important asymptomatic carriers of O157 STEC but can also be a reservoir of EPEC and eae positive non-O157 STEC, which are described in association with human diseases.

First isolation and further characterization of enteropathogenic Escherichia coli (EPEC) O157:H45 strains from cattle.

BACKGROUND: Enteropathogenic Escherichia coli (EPEC), mainly causing infantile diarrhoea, represents one of at least six different categories of diarrheagenic E. coli with corresponding distinct pathogenic schemes. The mechanism of EPEC pathogenesis is based on the ability to introduce the attaching-and-effacing (A/E) lesions and intimate adherence of bacteria to the intestinal epithelium. The role and the epidemiology of non-traditional enteropathogenic E. coli serogroup strains are not well established. E. coli O157:H45 EPEC strains, however, are described in association with enterocolitis and sporadic diarrhea in human. Moreover, a large outbreak associated with E. coli O157:H45 EPEC was reported in Japan in 1998. During a previous study on the prevalence of E. coli O157 in healthy cattle in Switzerland, E. coli O157:H45 strains originating from 6 fattening cattle and 5 cows were isolated. In this study, phenotypic and genotypic characteristics of these strains are described. Various virulence factors (stx, eae, ehxA, astA, EAF plasmid, bfp) of different categories of pathogenic E. coli were screened by different PCR systems. Moreover, the capability of the strains to adhere to cells was tested on tissue culture cells. RESULTS: All 11 sorbitol-positive E. coli O157:H45 strains tested negative for the Shiga toxin genes (stx), but were positive for eae and were therefore considered as EPEC. All strains harbored eae subtype alpha1. The gene encoding the heat-stable enterotoxin 1 (EAST1) was found in 10 of the 11 strains. None of the strains, however, carried ehx A genes. The capability of the strains to adhere to cells was shown by 10 strains harbouring bfp gene by localized adherence pattern on HEp-2 and Caco-2 cells. CONCLUSION: This study reports the first isolation of typical O157:H45 EPEC strains from cattle. Furthermore, our findings emphasize the fact that E. coli with the O157 antigen are not always STEC but may belong to other pathotypes. Cattle seem also to be a reservoir of O157:H45 EPEC strains, which are described in association with human diseases. Therefore, these strains appear to play a role as food borne pathogens and have to be considered and evaluated in view of food safety aspects.