Prevalence and characteristics of Shiga toxin-producing Escherichia coli, Salmonella spp. and Campylobacter spp. isolated from slaughtered sheep in Switzerland

Caecum samples collected from 653 slaughtered sheep from two Swiss abattoirs were examined. The aim of this study was: (i) to determine the prevalence of Shiga toxin-producing Escherichia coli (STEC), Salmonella spp. and Campylobacter spp.; (ii) to further characterize isolated strains; and (iii) to discuss the results obtained with their relevance to food safety. The percentage of samples testing positive for STEC by a polymerase chain reaction was 29.9%. The prevalence of positive Salmonella spp. samples was 11.0% and of Campylobacter spp. 17.5%. In 55.3% of the 76 isolated non-O157 STEC strains, stx2 variants (mostly stx2d) were detected. Additional virulence factors were harbored by 55.3% of the STEC strains, 10.5% of them being eae positive, 55.3% ehxA positive and 2.6% astA positive. All isolated salmonella were identified as Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7). Pulsed-field gel electrophoresis (PFGE) was performed for genotyping and 22 different restriction endonuclease digestion profiles were found among these strains for the different farms of origin. Of the 114 isolated Campylobacter spp. strains, 64.9% were shown to be Campylobacter jejuni and 35.1% Campylobacter coli, nine strains showed resistance against tetracycline, ciprofloxacin/nalidixic acid or streptomycin. In conclusion, sheep are a reservoir for the pathogens of latent zoonoses as non-O157 STEC, S. enterica subsp. diarizonae and Campylobacter spp. The maintenance of slaughter hygiene is consequently of crucial importance. It can be measured in daily practice by "slaughter-process-controls" and regular microbiological monitoring of carcasses. These are valuable tools for verifying slaughter hygiene according to hazard analysis critical control point principles.     

Incidence of coagulase positive Staphylococcus on beef carcasses in three Australian abattoirs.

The contamination of beef carcasses with coagulase-positive staphylococci (CPS) was studied at three beef abattoirs (A, B and C). The incidence and the number of CPS were determined on cattle hides immediately after slaughter and on three carcass sites (brisket, flank and round) at different points during processing along the slaughter line. The incidence of CPS on cattle hides ranged from 20 to 68.6%. At abattoir A, 6.5% of the carcasses sampled before evisceration were contaminated with CPS, compared to 40% of the carcasses after evisceration. The incidence on carcasses changed little during further processing; however, after chilling for 72 h, the incidence increased to 83%. After evisceration, the brisket and flank areas were more often contaminated than the round. A similar pattern of contamination was observed at abattoir B. At abattoir C, 26.7% of the samples collected before evisceration were contaminated and this fell to 16.7% after evisceration. After chilling for 72 h, the incidence of carcass contamination with CPS increased to 46.7%. The average number of CPS on contaminated carcasses prior to and after overnight chilling was less than 50 colony-forming units (cfu)/cm2 and, after weekend chilling, increased to 64 and 112 cfu/cm2 in abattoirs A and B, respectively. Of the isolates tested, 71.4% produced staphylococcal enterotoxin and 21% could not be classified phenotypically. The hands of workers and environmental sites associated with the evisceration process were examined for CPS at abattoir A. Hands were heavily contaminated and were the likely source of CPS contamination at this abattoir.     

Effects of shearing and fleece cleanliness on microbiological contamination of lamb carcasses

The meat industry in Norway has developed national guidelines for Good Hygiene Practices for slaughtering and skinning, based on categorisation of animals. These include shearing sheep and lambs in the abattoirs immediately before slaughter. The aim of this study was to investigate microbiological carcass contamination associated with: (i) different shearing regimes; (ii) fleece cleanliness; and (iii) the slaughter process. In addition, the efficacy of the national guidelines in reducing microbial contamination was evaluated. A total of 280 swab samples were collected from the brisket areas (100 cm(2)) of 140 naturally contaminated lamb carcasses in a commercial abattoir. Half the samples were collected at skinning of brisket areas at the start of the slaughter-line and half of them were collected at the end of slaughter-line, just before chilling. The lambs were divided into four groups (n=35) according to the duration of the period between shearing and slaughter: (i) 0 days (shorn at the abattoir immediately before slaughter); (ii) three days; (iii) seven days; and (iv) not shorn. Mean log colony forming units (CFU) per 100 cm(2) at skinning were 5.78 and 6.95 for aerobic plate count (APC) (P<0.05), 1.65 and 2.78 for Escherichia coli (P<0.05) for shorn and unshorn lambs, respectively. For shorn lambs, divided according to the period between shearing and slaughter, the mean log CFU per 100 cm(2) were 5.45, 5.75, 6.12 (APC) and 1.77, 1.46, 1.71 (E. coli) for the 0-days, 3-days and 7-days groups, respectively (P<0.05 for the difference between 0- and 7-days groups in APC results). A four-category scale (0-3) was used for assessing fleece cleanliness before skinning. Visually clean lambs (score '0') had lower levels of APC on the carcass surfaces than those categorised as dirty (score '2-3') (P<0.05). The carcasses at the end of the slaughter-line had lower levels of APC than they had at skinning. However, the statistical significant reduction of E. coli on carcass surfaces at skinning point for shorn lambs, were impaired and no longer significantly different from the unshorn group at the end of the slaughter-line. The increased E. coli level at the end of the slaughter-line might be explained by weaknesses related to slaughter hygiene in particular suboptimal evisceration in the abattoir which was used as a basis for our trial, and thus the national guidelines concerning shearing had not the fully intended effect on reducing microbial carcass contamination.     

Genotypic characterization of non-O157 Shiga toxin-producing Escherichia coli in beef abattoirs of Argentina

The non-O157 Shiga toxin-producing Escherichia coli (STEC) contamination in carcasses and feces of 811 bovines in nine beef abattoirs from Argentina was analyzed during a period of 17 months. The feces of 181 (22.3%) bovines were positive for non-O157 STEC, while 73 (9.0%) of the carcasses showed non-O157 STEC contamination. Non-O157 STEC strains isolated from feces (227) and carcasses (80) were characterized. The main serotypes identified were O178:H19, O8:H19, O130:H11, and O113:H21, all of which have produced sporadic cases of hemolytic-uremic syndrome in Argentina and worldwide. Twenty-two (7.2%) strains carried a fully virulent stx/eae/ehxA genotype. Among them, strains of serotypes O103:[H2], O145:NM, and O111:NM represented 4.8% of the isolates. Xba I pulsed-field gel electrophoresis pattern analysis showed 234 different patterns, with 76 strains grouped in 30 clusters. Nine of the clusters grouped strains isolated from feces and from carcasses of the same or different bovines in a lot, while three clusters were comprised of strains distributed in more than one abattoir. Patterns AREXSX01.0157, AREXBX01.0015, and AREXPX01.0013 were identified as 100% compatible with the patterns of one strain isolated from a hemolytic-uremic syndrome case and two strains previously isolated from beef medallions, included in the Argentine PulseNet Database. In this survey, 4.8% (39 of 811) of the bovine carcasses appeared to be contaminated with nonO157 STEC strains potentially capable of producing sporadic human disease, and a lower proportion (0.25%) with strains able to produce outbreaks of severe disease.     

Isolation and characterization of Shiga toxin-producing Escherichia coli O157:H7 and non-O157 from beef carcasses at a slaughter plant in Mexico

The contamination of beef carcasses with Shiga toxin-producing O157:H7 and non-O157 Escherichia coli (STEC) obtained from a slaughter plant in Guadalajara, Mexico was investigated. A total of 258 beef carcasses were sampled during a 12-month period. All samples were assayed for STEC by selective enrichment in modified tryptone soy broth supplemented with cefixime, cefsulodin and vancomycin, followed by plating on Sorbitol MacConkey Agar supplemented with cefixime and tellurite (CT-SMAC). Simultaneously, all samples were assayed by immunomagnetic separation (IMS) and plated on CT-SMAC and CHROMagar. The presence of the stx1, stx2, eaeA and hly933 genes, recognized as major virulence factors of STEC, was tested for O157:H7 and non-O157 E. coli isolates by multiplex polymerase chain reaction (PCR). STEC was detected in two (0.8%) samples. One of these STEC isolates corresponded to the serotype O157:H7 showing stx2, eaeA and hyl933 genes. The other isolate corresponded to non-O157 STEC and only had the stx1 gene. Thirteen carcasses (5%) were positive for nonmotile E. coli O157 and 7 (2.7%) were positive for E. coli O157:H7. The presence of O157:H7 and non-O157 STEC on beef carcasses in this slaughter plant in Guadalajara, Mexico, emphasizes the importance of implementing the Hazard Analysis and Critical Control Point (HACCP) system, as well as the need for implementing, evaluating, and validating antimicrobial interventions to reduce the presence of potential pathogenic microorganisms.     

Number and distribution of bacteria on some beef carcasses at selected abattoirs in some member states of the European communities

In seven member countries of the European Communities, three abattoirs were visited on three occasions in each of two surveys and at each visit ten beef carcasses were sampled, before chilling, at defined sites on the neck, brisket, forerib and medially on the round. In Survey I, samples were plated for total viable count (TVC) at 30° (ISO 2293) and Enterobacteriaceae at 37° (ISO 5552); in Survey II only TVCs were made. This paper is confined to analyses of the TVCs in the two surveys. Data from each country were analysed separately as sampling methodology may not have been sufficiently reproducible by different workers to allow between-countries comparison. Variations among visits to particular abattoirs and abattoir × site interactions made comparisons among abattoirs invalid within five of the seven countries. To effectively monitor differences between abattoirs within most countries it would be necessary to make more than three visits to each abattoir. Despite abattoir × site interactions in three countries in Survey I and four countries in Survey II, comparisons between sites were generally valid because of the consistent high contamination of the brisket. In the remainder of countries the abattoir × site interaction was too large to allow valid comparisons between sites. It is recommended that at least three or four sites are sampled in future surveys as only one site per carcass would underestimate the number of more heavily contaminated carcasses.     

Contamination of bovine carcasses and abattoir environment by Escherichia coli O157:H7 in Istanbul

The aim of this study was to investigate contamination of carcasses and abattoir environment with Escherichia coli O157:H7. Five abattoirs in Istanbul were visited between January 2000 and April 2001. During visits, sampling was performed and a total of 330 cattle were selected. Cattle were examined for the presence of faeces on the hide (abdomen and legs) before slaughter. The swabs from the carcasses and environmental samples (abattoir floor, benches including conveyors, knives, aprons, saws, hooks, hands) were taken at the abattoir immediately after slaughter using sterile cotton swabs. A sample from the wash water of the abattoir was also taken. Preenrichment, immunomagnetic separation and CT-SMAC agar were used for the isolation. The reaction of the isolates with anti-O157 and H7 antisera were also analysed. Twelve strains (3.6%) of E. coli O157 were isolated from the cattle carcasses and eight (2.4%) of them gave positive reaction with anti-H:7. Six strains of E. coli O157 were isolated from the environmental samples and all strains were positive for H7. The number of E. coli O157H:7 strains isolated from the environmental samples was two from the knife, two from the hands, one from the apron and one from the floor. No E. coli O157 was isolated from the abattoir water.     

Microbiological sampling of swine carcasses: a comparison of data obtained by swabbing with medical gauze and data collected routinely by excision at Swedish abattoirs

Swab sample data from a 13-month microbiological baseline study of swine carcasses at Swedish abattoirs were combined with excision sample data collected routinely at five abattoirs. The aim was to compare the numbers of total aerobic counts, Enterobacteriaceae, and Escherichia coli, recovered by swabbing four carcass sites with gauze (total area 400 cm2) with those obtained by excision at equivalent sites (total area 20 cm2). The results are considered in relation to the process hygiene criteria that are stated in Commission Regulation (EC) No 2073/2005. These criteria apply only to destructive sampling of total aerobic counts and Enterobacteriaceae, but alternative sampling schemes, as well as alternative indicator organisms such as E. coli, are allowed if equivalent guarantees of food safety can be provided. Swab sampling resulted in higher mean log numbers of total aerobic counts at four of the five abattoirs, compared with excision, and lower or equal standard deviations at all abattoirs. The percentage of swab and excision samples positive for Enterobacteriaceae at the different abattoirs ranged from 68 to 100% and 15 to 24%, respectively. Similarly, the percentages of swab samples that were positive for E. coli were higher than the percentages of positive excision samples (range 52 to 84% and 3 to 14%, respectively). Due to the low percentage of positive excision results, the mean log numbers of Enterobacteriaceae and E. coli were only compared at two and one abattoirs, respectively, using log probability regression to substitute censored observations. Higher mean log numbers of Enterobacteriaceae were recovered by swabbing compared with excision at one abattoir, whereas the numbers of Enterobacteriaceae and E. coli did not differ significantly between sampling methods at one abattoir. This study suggests that the same process hygiene criteria as those stipulated for excision can be used for swabbing with gauze without compromising food safety. For monitoring of low numbers of Enterobacteriaceae and E. coli, like those found on swine carcasses at Swedish abattoirs, the results also show that swabbing of a relatively large area is superior to excision of a smaller area.     

Microbiological contamination of pig carcasses at different stages of slaughter in two European Union-approved abattoirs

At sequential steps of slaughter (scalding, dehairing, singeing, polishing, trimming, washing, and chilling), 200 pig carcasses from two abattoirs were examined for total viable bacteria count (TVC) and the presence of Enterobacteriaceae and coagulase-positive Staphylococcus (CPS) by the wet-dry double-swab technique at the neck, belly, back, and ham. Before scalding, mean TVCs ranged from 5.0 to 6.0 log CFU cm(-2), and Enterobacteriaceae and CPS were detected on all carcasses. At abattoir A, mean TVCs and the percentage of Enterobacteriaceae-positive carcasses were reduced (P < 0.05) after scalding (1.9 log CFU cm(-2) and 12%, respectively), singeing (1.9 log CFU cm(-2) and 66%, respectively), and blast chilling (2.3 log CFU cm(-2) and 17%, respectively) and increased (P < 0.05) after dehairing (3.4 log CFU cm(-2) and 100%, respectively) and polishing (2.9 log CFU cm(-2)). The proportion of CPS-positive samples decreased to < or = 10% after scalding and remained at this level. At abattoir B, mean TVCs and the percentages of Enterobacteriaceae- and CPS-positive carcasses were reduced (P < 0.05) after scalding (2.4 log CFU cm(-2) and 29 and 20%, respectively), polishing (3.7 log CFU cm(-2)), and chilling (2.6 log CFU cm(-2) and 55 and 77%, respectively) and increased (P < 0.05) after the combined dehairing-singeing (4.7 log CFU cm(-2) and 97 and 100%, respectively). Among sites, the neck tended to yield higher levels of contamination from trimming to chilling at both abattoirs (P < 0.05). Consequently, scalding, singeing, and chilling may be integrated in a hazard analysis critical control point (HACCP) system for pig slaughter. As indicated by the higher levels of contamination on carcasses after dehairing-singeing and the following stages at abattoir B, each abattoir should develop its own baseline data and should customize HACCP systems to match process- and site-specific circumstances.     

The effect of abattoir design on aerial contamination levels and the relationship between aerial and carcass contamination levels in two Irish beef abattoirs

This study investigated the relationship between aerial and beef carcass contamination and examined the effect of abattoir design and time of slaughter on the aerobiology of slaughter operations in two commercial beef abattoirs. A dual head impaction air sampler and swab samples taken from 100 cm² of the brisket of beef carcasses, were used to examine Total Viable, Psychrotrophic, Enterobacteriaceae and Pseudomonad numbers. In Abattoir A, with a straight-line single-floor design, airborne bacterial numbers were generally lower in the “clean” than in the “dirty” area of the plant. In Abattoir B, which had a serpentine two-floor design, this trend was generally reversed. Both abattoirs displayed a similar pattern in airborne counts over the production day, with numbers generally being lower before slaughter, than in the morning and afternoon. Correlations between aerial and carcass contamination for each of the bacterial groups on the slaughter line in Abattoirs A and B were poor. The data suggest that it is difficult to make a definitive evaluation of the relationship between aerial and carcass contamination levels. Methods currently used to determine the relationship between aerial and carcass contamination need to be reconsidered.     

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.     

Using indicator bacteria and Salmonella test results from three large-scale beef abattoirs over an 18-month period to evaluate intervention system efficacy and plan carcass testing for Salmonella

To develop a process for predicting the likelihood of Salmonella contamination on beef carcasses, we evaluated the influence of several possible causative factors (i.e., year, abattoir, day of week, month, and intervention system components) on the risk of Salmonella and indicator organism contamination. Hide and carcass sponge samples were collected in 2005 to 2006 in six steps at three abattoirs in the East (A), Midwest (B), and Southwest (C) United States. Each abattoir used the same intervention system. Samples were analyzed for aerobic plate counts (APCs; n = 18,990) and Enterobacteriaceae counts (EBCs; n = 18,989) and the presence or absence of Salmonella (n = 5,355). Our results demonstrated that many factors play a significant role in the level of microbial contamination of beef carcasses. Overall, Salmonella prevalence and EBC levels were significantly higher in 2006 than in 2005. APCs and EBCs were highest in abattoirs A (3.57 log CFU/100 cm2) and B (1.31 log CFU/100 cm2). The odds of detecting a positive Salmonella isolate were greatest in abattoir C and lowest in abattoir A. Across the three abattoirs, the overall intervention process effectively reduced microbiological contamination. Salmonella prevalence fell from 45% (preevisceration) to 0.47% (postchilled-lactic acid), and there were APC and EBC reductions of 5.43 and 5.28 log CFU/100 cm2, respectively, from hide-on to postchilled-lactic acid samples. At each abattoir, composites of three individual EBC-negative carcass samples yielded Salmonella-negative results 97 to 99% of the time. These results suggest the possibility of using indicator test results to accurately predict the absence of Salmonella in a beef carcass sample.     

Determination of the principal points of product contamination during beef carcass dressing processes in Northern Ireland

To determine the principal points of microbial contamination of carcasses during beef carcass dressing in Northern Ireland, 190 carcasses were sampled by swabbing 1,000 cm2 of the brisket. A detailed survey of one abattoir was initially conducted, with sampling of a total of 100 carcasses immediately after hide removal (H), after carcass splitting (S), and immediately after washing (W) before dispatch to the chiller. The total bacterial counts after incubation at both 22 and 37 degrees C indicated that there was no significant increase in the numbers of bacteria after the first sampling point, H (P > 0.05). To determine whether this was the case in the majority of Northern Ireland abattoirs, 15 carcasses were then sampled at each of an additional six abattoirs, at points H and W only. Total bacterial counts were significantly higher (P < 0.05) at H than at W, indicating that hide pulling was the major point of bacterial contamination of beef carcasses and hence a critical control point for the final microbiological quality of the carcasses. Mean counts of Enterobacteriaceae at both incubation temperatures were very low (< 10 CFU/cm2) but were higher at W than at H, probably indicating that washing was redistributing bacteria from the posterior to the anterior region.     

Microbiological contamination of pig and cattle carcasses in different small-scale Swiss abattoirs

A total of 750 pig carcasses and 535 cattle carcasses from 17 small-scale abattoirs were sampled by excision at four sites (pig: neck, belly, back, ham; cattle: neck, brisket, flank, rump). Samples were examined for total viable counts (TVC) and Enterobacteriaceae. Mean TVCs ranged from 2.4 to 4.2 log(10)CFUcm(-2) on pig carcasses and from 2.7 to 3.8 log(10)CFUcm(-2) on cattle carcasses. With regard to EU Regulation (EC) No 2073/2005, TVCs were mainly considered satisfactory (pig: 81.3%; cattle: 71.4%). Amongst sites, the back (pigs) and neck (cattle) tended to yield higher TVCs. Enterobacteriaceae were detected in low counts on 23.9% of pig carcasses and 21.7% of cattle carcasses. Amongst abattoirs, Enterobacteriaceae prevalence on pig and cattle carcasses ranged from 2.0% to 56.0% and from 0.0% to 55.0%, respectively. Consequently, criteria of the EU Regulation proved to be a suitable tool for the appraisal of microbiological results (TVCs) from pig and cattle carcasses from small-scale abattoirs. Because the occurrence of Enterobacteriaceae on carcasses was too infrequent to ensure log normality, frequencies should be compared for these organisms.     

Assessment of carcass contamination with E. coli O157 before and after washing with water at abattoirs in Nigeria

The study was carried out to assess the level of beef carcass contamination with Escherichia coli including O157 strains before and after washing with water. Samples of water used for washing carcasses were collected and thirty beef carcasses were swabbed within a period of one month in each of three abattoirs located in North-Western states of Nigeria. E. coli were enumerated as indicator organisms. Using conventional biochemical tests, the isolation rate of E. coli in the 120 swab samples collected in each abattoir from external and internal surfaces of the carcasses was 58.3% at Kano abattoir, 70.8% at Sokoto abattoir, while 76.7% was recorded at Zango abattoir. E. coli counts from external and internal surfaces of the carcasses were enumerated as mean log and ranged between 4.3 Log(10) and 4.6 Log(10) cfu/cm(2) before washing, while the values were 4.6 Log(10) and 4.9 Log(10) cfu/cm(2) after washing. Data analysis revealed that the increase in E. coli counts after washing carcasses with water was statistically significant (P<0.05) in all the abattoirs. However, there was no statistically significant difference (P>0.05) between the 3 abattoirs in mean log of E. coli counts from external surfaces of carcass after washing. E. coli O157 was identified from both the water and surfaces of carcasses using Latex agglutination kit. A prevalence of 2.8% of E. coli O157 was detected in 360 swab samples from 90 beef carcasses examined. E. coli counts from water used in washing carcasses were between 22 and 120 cfu/100 ml. Of the 72 water samples, 3(4.2%) were positive for E. coli O157. In conclusion, there was increased contamination of carcasses during processing and water used in washing carcasses might have contributed to carcass contamination in all the abattoirs studied due to use of non-potable water.     

Characteristics of Shiga toxin-producing Escherichia coli from meat and milk products of different origins and association with food producing animals as main contamination sources

Shiga toxin-producing strains of Escherichia coli (STEC) cause diarrhoea and haemorrhagic colitis in humans. Most human infections are attributed to consumption of STEC contaminated foodstuff. Food producing animals constitute important reservoirs of STEC and serve as source of food contamination. In this study, we have analyzed 593 foodborne STEC strains for their serotypes and for nine virulence genes (stx1, stx1c, stx1d, stx2, stx2b, stx2e, stx2g, E-hly and eae). The 593 STEC strains grouped into 215 serotypes, and 123 serotypes (57.2%) were represented each by only one STEC isolate. Fifteen serotypes (7.0%) were attributed to 198 (33.3%) of the 593 STEC strains. The foodborne STEC were grouped into different categories in relation to the species of the food producing animal (cattle, pigs, sheep, goats, red deer, wild-boar and hare). Univariate and multivariate statistical analyses revealed significant similarities between the animal origin of the food and the virulence markers of foodborne STEC. Significant associations (p < 0.001) were found for stx1 and for stx2 with bovine meat and milk products. The stx2e gene was significantly (p < 0.001) associated with STEC from pork and wild boar meat. Stx1c and stx2b genes were significantly (p < 0.001) more frequent in STEC from deer meat, as well as from meat and milk products derived from sheep and goats. Using logistic regression models we detected significant (p < 0.01) combinations between stx1, stx2 and E-hly genes and STEC from bovine meat. The combination of stx1c and stx2b genes was significant (p < 0.001) for STEC derived from red deer, sheep and goat products. The properties of foodborne STEC were compared with published data on faecal STEC from food producing animals. Virulence profiles and serotypes of STEC from food showed remarkable similarities to those of faecal STEC that were from the same animal species. The findings from our study clearly indicate that the food producing animals represent the most important source for the entry of STEC in the food chain. Sound hygiene measures implemented at critical stages of food production (milking, slaughtering, and evisceration) should be most effective in reducing the frequency of STEC contamination of food derived from domestic and wildlife animals.     

The development of a 'clean sheep policy' in compliance with the new Hygiene Regulation (EC) 853/2004 (Hygiene 2)

The aim of this research was to identify the risk factors associated with the transfer of bacterial contamination from the fleece to the ovine carcass thereby providing the scientific basis for the development and validation of a clean sheep policy. Two hundred sheep in lairage were graded into five categories each consisting of 40 sheep. The categories were as follows; (A) clean and dry; (B) clean and wet; (C) dirty and dry; (D) dirty and wet and (E) visible dags (dung-clotted tufts of wool) categorized by the chief veterinary inspector at the slaughter plant based on the visual inspection of the hygienic status of the fleece. Microbiological evaluations of the carcasses were conducted using swab sampling methods. Total viable counts (TVCs), Enterobacteriaceae and coliform counts were obtained from 40 animals per category at four separate sites (brisket, shoulder, flank and rump) immediately after pelt removal. Statistical analysis of TVC data obtained from the carcass indicated that the dirt level of the fleece had a significant effect on contamination levels when the fleece was dry. Enterobacteriaceae and coliform counts suggest that dirt was a contributing risk factor regardless of wetness or dryness of the animal. The clean sheep policy should therefore differentiate between clean and dirty sheep and mandate additional hygiene measures for the latter.     

An alternative approach for enumeration of Escherichia coli in foods

A study was carried out in France in collaboration with the meat industry to investigate the occurrence and characteristics of Shiga toxin-producing E. coli (STEC) and O157 E. coli in a population of healthy bovines representative of French livestock. A total of 851 animals belonging to three bovine classes (106 young bulls, 374 dairy cows and 371 meat cows) were included in the study. Samples of feces and of the corresponding carcasses were collected from March 97 to August 97 in seven abattoirs spread throughout the national territory. STEC cultures from the 1702 samples were screened using PCR for the presence of stx genes. Positive samples were further subjected to colony blot hybridization and to O157-specific immunomagnetic separation. Probe-positive colonies and O157 colonies were then analyzed for the presence of virulence genes and phenotypic characters (serotype, Stx production). In 154 (18.1%) feces and 91 (10.7%) carcass samples stx genes were detected. Two hundred and twenty-two STEC colonies were isolated from 67 (7.9%) feces and 16 (1.9%) carcass samples, with 183 STEC isolated from feces and 39 from carcasses. Only eight O157 isolates were collected from feces samples. None of these O157 E. coli isolates presented stx genes and thus could not be considered as pathogenic regarding hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). In 3.2% of STEC isolated from feces and in 10.2% of STEC from carcasses eae genes were detected. In 17% of STEC from feces and in 30.7% from carcasses ehx genes were detected. Using these data, the 222 STEC colonies could be classified in 11 different 'virulence patterns' (presence/absence of stx1, stx2, eae and ehx genes), showing that more than 77% of isolates presented only one virulence factor. Only three STEC on 222 colonies (1.3%) presented the three virulence factors stx, eae and ehx in association, none of them reacting with antisera specific for enterohemorrhagic E. coli. (EHEC). These data, together with the fact that only five isolates on the 222 (2.2%) reacted with such antisera (three O111 and two O26 isolates) demonstrated that the natural bacterial populations isolated during this study were clearly distinct from EHEC.     

Microbiological contamination of cattle and pig carcasses at five abattoirs determined by swab sampling in accordance with EU Decision 2001/471/EC

A total of 800 cattle carcasses (abattoir A: n=200; B: n=150; C: n=150; D: n=150, E: n=150) and 650 pig carcasses (abattoir A: n=200; B: n=150; C: n=150; D: n=150) were examined at five Swiss abattoirs with an annual slaughtering capacity >10 million kg. Weekly, 10 cattle and 10 pig carcasses were sampled at four sites by the wet-dry double swab technique. From each carcass the samples were pooled and examined for total viable counts (TVC) and Enterobacteriaceae. At the abattoirs, mean log TVCs from cattle carcasses ranged from 2.1 to 3.1 cm(-2) and those from pig carcasses from 2.2 to 3.7 cm(-2). Daily TVC mean log values showed significant differences between abattoirs (P<0.05), whereas no significant differences were detected between animal species. On average, Enterobacteriaceae were detected (i) in low counts, (ii) on 31.0% of cattle and on 20.2% of pig carcasses, and (iii) more frequently and in higher counts on cattle than on pig carcasses (P<0.05). Data from this study indicate that the wet-dry double swab technique is suitable to determine microbiological contamination of cattle and pig carcasses in accordance with EU Decision 2001/471/EC. For samples obtained by the non-destructive technique from cattle and pig carcasses, the following microbiological performance criteria are proposed: Daily mean log values are acceptable, marginal, and unacceptable for TVC when they are <3.00, 3.00-4.00, >4.00 cm(-2), and for Enterobacteriaceae when they are <1.00, 1.00-2.00, and >2.00 cm(-2). However, such values have to be seen merely as baselines. It is important to implement a monitoring system based on abattoir-specific data and criteria as permitted by quality control chart methods.     

Prevalence and Concentration of Arcobacter spp. on Australian Beef Carcasses

The International Commission on Microbiological Specifications for Foods (ICMSF) classified Arcobacter spp. as emerging pathogens in 2002. Arcobacter spp. have been isolated from numerous food products at retail and from animal carcasses and feces at slaughter. A survey was conducted to determine both the prevalence and concentration of Arcobacter spp. on prechill beef carcasses. Surface swab samples were collected from 130 beef carcasses at the end of processing, prior to chilling. The concentration of Arcobacter spp. was determined by a most-probable-number per square centimeter (3 by 3) method with a limit of detection of 0.12 CFU/cm(2). Of the 100 carcasses examined from export abattoirs, 20 (20.0%) were contaminated with Arcobacter spp., and 5 of these had quantifiable levels of contamination ranging from 0.12 to 0.31 CFU/cm(2). Of the 30 carcasses examined at a pet food abattoir, 25 (83.3%) were contaminated with Arcobacter spp., and 10 of these had quantifiable levels of contamination ranging from 0.12 to 0.95 CFU/cm(2). Three species of Arcobacter, A. butzleri, A. cryaerophilus, and A. skirowii, were identified by PCR. Each of the species was present in an approximately equal ratio from export abattoirs. This study demonstrates that slaughter practices at export abattoirs are sufficient to maintain both low prevalence and low levels of contamination of beef carcasses with Arcobacter spp.