Prevalence of Yersinia enterocolitica in fattening pigs

The prevalence of pathogenic Yersinia enterocolitica in pig herds was monitored during six trials (at four different farrow-to-finisher farms). Samples were taken throughout the whole rearing period from birth of the piglets to the final fattening stage, and different samples were taken from these pigs during the slaughter process. Environmental samples also were evaluated to identify potential sources of on-farm infection. Y. enterocolitica was isolated using irgasan-ticarcillin-potassium chlorate broth enrichment and cefsulodin-irgasan-novobiocin agar culture. Colonies were identified using bio- and serotyping methods and by PCR assay. Pathogenic Y. enterocolitica were not isolated from fecal samples from piglets and weaners. The only fecal samples positive for Y. enterocolitica were obtained during the fattening stage. The prevalence of Y. enterocolitica in fattening pig herds ranged between 0 and 65.4%. Y. enterocolitica isolates were detected at the abattoir in 38.4% of the tonsils, in 3.8% of the ileocecal lymph nodes, on 0.3% of the carcass surfaces before chilling, and on 0% of the carcass surfaces after chilling. Almost all isolates belonged to bioserotype 4/O:3. Only one strain was identified as O:9. All isolates contained the ail gene. The yopT gene was found in 99.1% of the farm isolates but in only 76.6% of the isolates found at the abattoir from the corresponding carcasses. Although a direct link between porcine isolates and human infection has not been demonstrated, the similarity of the bioserotypes in infected pigs and humans and the presence of virulence factors in porcine isolates should encourage further studies to determine the risk of transmission of Y. enterocolitica to humans from pigs and pork products.     

Contamination of freshly slaughtered pig carcasses with human pathogenic Yersinia enterocolitica

Evidence is presented for the extent of contamination of freshly slaughtered pig carscasses with human pathogenic Yersinia enterocolitica and shows the significance of faecal contamination as a source of infection. Swab samples collected from the rectum and the surface of a total of 1458 pig carcasses were examined for the presence of human pathogenic Y. enterocolitica Y. enterocolitica, biovar IV, serogroup 0:3, were isolated from the rectum of 360 pigs (24.7%). The organism was isolated from carcass surfaces with varying frequencies depending on the evisceration technique. Manual evisceration was found to correspond with high frequencies of contamination: 26.3% on the medial hind limb and 12.9% on the split sternum. The use of a mechanised bung cutter was found to reduce the rate of contamination, especially when the bung cutter was used in connexion with enclosing the anus and rectum in a plastic bag to minimise faecal contamination. When carcasses were eviscerated in this way, it was possible to reduce carcass contamination to 1.9% on the medial hind limb, 1.0% in the pelvic duct, and 2.2% on the split sternum.     

Slaughter pigs and pork as a source of human pathogenic Yersinia enterocolitica.

Pathogenic Yersinia enterocolitica strains (serogroups 0:3;0:9 and 0:5.27) were isolated from 36 (42%) of 86 porcine tonsils, 8 (20%) of 40 tongues, 17 (17%) of 100 rectal swabs and from 4 (1%) of 400 pork samples. Pathogenic Yersinia strains were not isolated from samples of 210 pig carcasses and from 20 samples of porcine head meat. These results confirm that pigs are an important reservoir of pathogenic Y. enterocolitica. However, contamination of carcasses during the slaughtering process with Yersinia from either faecal material or from the tonsillary region does not seem to occur frequently and this may also explain the low contamination rate of pathogenic Y. enterocolitica found for pork. For the isolation of pathogenic Y. enterocolitica strains from foods, enrichment in irgasan-ticarcillin-chlorate broth (ITC) and isolation on SS-deoxycholate-calcium agar (SSDC) is recommended.     

Prevalence of Yersinia enterocolitica in market weight hogs in the United States

Pigs are the major animal reservoir for Yersinia enterocolitica strains, which are potentially pathogenic for humans. The goals of this study were (i) to estimate the individual animal and on-farm prevalences of Y. enterocolitica in hogs based on tonsil samples collected during National Animal Health Monitoring System Swine 2002 study and (ii) to use these data with data previously published for fecal samples to determine on-farm risk factors for Y. enterocolitica. Tonsil swabs (1,218) and fecal samples (2,847) were collected on 124 farms located in the top 17 pork-producing states. Ten percent of tonsils (122 of 1,218 samples) were positive in irgasan-tiracillin-chlorate (ITC) enrichment broth by real-time PCR, but only 5.6% of samples (68 of 1,218) were positive after subculture on the more selective cefsulodin-irgasan-novobiocin (CIN) agar. For tonsils, the on-farm prevalence based on real-time PCR detection of the ail gene in ITC enrichment broth cultures was 32% (32 of 100 premises sampled); the prevalence based on subculture in CIN agar was 19.6% (20 of 102 premises). Results of bacteriological isolation and real-time PCR analysis of tonsils and feces were combined to estimate prevalence (individual animal and farm), which was subsequently correlated with 40 farm management practices. Four factors and their accompanying odds ratios (ORs) were identified in the final regression model: location in a central state (OR = 0.3), vaccination for Escherichia coli (OR = 3.0), percentage of deaths due to scours (OR = 3.5), and presence of meat or bone meal in grower-finisher diet (OR = 4.1).     

Detection of Salmonella spp., Yersinia enterocolitica and verocytotoxin-producing Escherichia coli O157 in pigs at slaughter in Italy

From December 1999 to December 2000, 150 pigs were randomly selected in two large abattoirs of northern Italy. Caecal material and carcass swabs were collected and examined for Salmonella, Yersinia enterocolitica, and Escherichia coli O157. Tonsils were examined for Salmonella and Y. enterocolitica. Salmonella was isolated from the intestinal content of 55 (36.7%) specimens, from 8 (5.3%) tonsils, and from 9 (6.0%) carcasses. Ten different serotypes were detected; the more common were Salmonella derby (37.8%), Salmonella bredeney (21.6%), and Salmonella typhimurium (14.8%). S. typhimurium isolates that belonged to phage-types DT104 and DT208 were 45% and 27.3%, respectively; 18.2% belonged to U302 and 9.1% were non-typeable. Y. enterocolitica was detected in the intestinal matter of 6 (4.0%) slaughtered pigs and in 22 (14.7%) tonsils; however, this pathogen was not found on carcasses. The majority of Y. enterocolitica isolates (82.1%) belonged to serotype O:3 biotype 4, one (3.6%) belonged to serotype O:9, and 13% did not belong to any known biotype. Verocytotoxin-producing E. coli (VTEC) O157 was isolated from the intestinal content of one (0.7%) slaughtered pig and from one (0.7%) carcass; four (2.7%) faecal samples contained E. coli O157 strains negative for the presence of both eae and VT genes.     

Yersinia enterocolitica in food hygiene

Yersinia enterocolitica and Yersinia enterocolitica-like bacteria constitute a fairly heterogenous group of bacteria which includes both well-established pathogens and a range of environmental strains which are ubiquitous in terrestrial and freshwater ecosystems. Pathogenic significance in man is mainly associated with a few serogroups (O:3, O:9, O:8, O:5,27). The pathogenic serogroups show different geographical distributions. The development of isolation procedures which clearly differentiate pathogenic from non-pathogenic variants has been difficult. Of special significance in food hygiene is the ability of Y. enterocolitica to grow in refrigerated foods. There is strong indirect evidence that pigs and food products of porcine origin are the major sources for human infection with Y. enterocolitica serogroups O:3 and O:9, the dominant human pathogens in most parts of the world. The reservoir(s) for serogroup O:8, which prevails in the U.S.A., is uncertain. The pig is the only animal consumed by man which regularly harbours pathogenic Y. enterocolitica. Improved isolation methods and DNA colony hybridization using genetic probes has indicated that the prevalence of pathogenic Y. enterocolitica in pork products is substantially higher than previously suggested. Prevention and control measures should focus on information of people involved in food processing and preparation and on the improvement of hygiene during slaughtering of swine. Important critical control points at the stage of slaughter are: (i) circumanal incision and removal of intestines, (ii) excision of the tongue, pharynx, and particularly the tonsils, (iii) post-mortem meat inspection procedures which involve incision of the mandibular lymph nodes, and (iv) deboning of head meat.     

Differences in heat resistance among pathogenic Yersinia enterocolitica depended on growth temperature and serotype

To gain a better understanding about the effect of growth temperature on heat resistance of Yersinia enterocolitica, we determined decimal reduction times at 60 degrees C (D60-values) for O:3; O:5,27; O:8; and O:9 strains harboring virulence plasmid coding for Yersinia outer membrane protein and experimentally virulence plasmid-deleted strains after they were grown to stationary phase at 7, 25, or 37 degrees C. Bacteria were inoculated into Trypticase soy broth and were incubated at several temperatures. D60-values of O:3; O:5,27; and O:8 strains were larger when they were grown at 37 degrees C than at 7 or 25 degrees C, despite the presence or absence of virulence plasmids. However, similar D60-values were observed in O:9 strains, despite growth at 7, 25, or 37 degrees C. The results indicate two types of Y. enterocolitica strains, growth temperature-dependent and -independent, and a Yersinia outer membrane protein that is not directly involved in growth temperature-dependent heat resistance.     

Testing of pathogenic Yersinia enterocolitica in pig herds based on the natural dynamic of infection

This study was performed to evaluate testing methods of pathogenic Yersinia enterocolitica in pigs at different ages. Relevant tools and procedures are crucial if pig herds should be declared free from pathogenic Y. enterocolitica. Historical data based on serology showed that the two farms investigated in this study (herds A and B) were contaminated with Y. enterocolitica O:3 since at least 1995. Laboratory investigations of 60 pigs were sampled one to four times (herd A) and 20 pigs were sampled one to three times (herd B) at different ages were the basis for this report. The following testing procedures could be used to conclude that a herd is free from pathogenic Y. enterocolitica:--serological testing of pigs could be performed as a basis for categorisation for all ages from about 100 days including at slaughter when the pigs are 150-180 days old, --bacteriological examination of faeces could be used as a basis for categorisation at all ages from 85 days until about 135 days, --bacteriological examination of tonsils could be used as a basis for categorisation at all ages from 85 days including at slaughter when the pigs are 150-180 days old. However, due to animal welfare aspects, one should avoid sampling of tonsils. Accordingly, the serological method or bacteriological examination of faeces at relevant ages should be preferred. One aspect related to slaughter hygiene is that in pigs slaughtered at the age of 135 days or more, the tonsils may be a more significant source of human pathogenic Y. enterocolitica than faeces.     

Occurrence of pathogenic Yersinia enterocolitica in Norwegian pork products determined by a PCR method and a traditional culturing method

From October 1997 to April 1998, a survey was conducted to assess the occurrence of pathogenic Yersinia enterocolitica in Norwegian pork products, using a traditional culturing method and a PCR assay. A total of 300 pork samples was examined. Five slaughterhouses in the Norwegian Meat Cooperative were represented with 249 samples and another 51 samples were obtained from retail outlets in the city of Oslo. Using the NMKL method, Y. enterocolitica 0:3 was isolated from six (2%) of the samples, while the PCR method indicated presence of pathogenic Y. enterocolitica in 50 (17%) of the samples. The results indicate that a reduction has occurred in the prevalence of pathogenic Y. enterocolitica in Norwegian pork products, as compared to a previous Norwegian study conducted in 1988-1989. The study also highlights the need for further development and improvement of methods applied for the detection of pathogenic Y. enterocolitica in foods.     

Prevalence of virulent Yersinia enterocolitica in bulk raw milk and retail cheese in northern-west of Iran

To determine the prevalence of virulent Yersinia enterocolitica, 554 samples consisting of 354 bulk raw milks and 200 traditional cheeses were collected from different parts of Eastern-Azerbaijan province, during a 23-month period from 2008 to 2010. The occurrence of virulent strains of Y. enterocolitica in samples enriched in peptone sorbitol bile broth (PSBB) was evaluated via the detection of attachment invasion locus (ail) gene by PCR. The viability of virulent Y. enterocolitica in the PCR-positive samples was tested using conventional culture method and the isolates were confirmed by the second-phase ail-PCR. According to the results, 8.66% of total samples including 7.62% of bulk raw milks and 10.5% of raw milk cheeses were found ail-positive by PCR method; subsequently Y. enterocolitica was isolated by the culture method and confirmed by the second phase ail-PCR in 2.88% of total samples including 2.26% of raw milks and 4% of cheese samples. It was concluded that, a sample enrichment followed by ail-PCR was more sensitive and robust to detect and distinguish the virulent strains of Y. enterocolitica compared to the conventional culture method.     

Prevalence and antimicrobial susceptibility of Escherichia coli O157:H7 on beef cattle slaughtered in Amman abattoir

Cattle are the main asymptomatic reservoir of Escherichia coli O157:H7 which can cause illness to human. The objectives of the study were to measure the prevalence of E. coli O157:H7 on cattle slaughtered in Amman abattoir, detect virulence factors in the isolates, determine antibacterial resistance of the isolates, and know how the isolates are different or similar when compared to characterized isolates from developed countries.     

Antimicrobial resistance of Yersinia enterocolitica strains from human patients, pigs and retail pork in Switzerland

To obtain basic data for future resistance monitoring programs, 386 Yersinia enterocolitica strains from human patients, raw retail pork and pig feces were tested for their susceptibilities to 16 antimicrobial agents and two antimicrobial growth promoters (carbadox and olaquindox). No strains were resistant to ceftriaxone, cefuroxime, ciprofloxacine, gentamicin, kanamycin, neomycin or polymyxin. Although in Switzerland carbadox and olaquindox were used as growth promoters for pigs for over 25 years, all strains were susceptible to them. In contrast, there were high levels of resistance to ampicillin, cefalothin and amoxicillin/clavulanic acid. Less than 10% of clinical isolates and strains from pig feces were resistant to streptomycin, sulfonamide, trimethoprim/sulfamethoxazole, tetracyclin, trimethoprim and chloramphenicol, but strains from retail pork were all susceptible to these antimicrobial agents. This finding suggested that pork is probably not a major source of Y. enterocolitica that cause human infections in Switzerland. A difference between clinical isolates and strains from pork was also shown by serotyping. Clinical isolates frequently belonged to the O3 and O9 groups whereas these two serotypes were not found in strains from pork. Resistance to multiple antimicrobial agents was rare. When examined by pulsed field gel electrophoresis (PFGE), two strains of fecal origin with an identical pattern of resistance to six antimicrobial agents were shown to be unrelated. Of four clinical isolates with resistances to five antimicrobial agents, two were of the same pulsotype. Retrospectively, it was found that these strains came from two members of the same household and thus represented a mini-outbreak.     

新等温扩增技术检测小肠结肠炎耶尔森氏菌

应用新型等温扩增检测技术——交叉引物等温扩增结合免疫金标试纸条建立检测小肠结肠炎耶尔森氏菌的方法。针对小肠结肠耶尔森氏菌16-23S rDNA间区序列设计特异性引物及探针,用54株小肠结肠炎耶尔森氏菌及相近株细菌进行特异性试验;通过纯菌液计数、样品中添菌检测进行灵敏度验证;对677份食品用传统生化国标法进行比较检测试验。建立方法具有较好特异性;增菌液检测灵敏度为10~1cfu/mL,当每25g样品中有10~0cfu菌时经增菌步骤后即可检出,样品检测同传统检测结果比较大致相符,没有漏检,假阳性率较低。建立的新型恒温检测方法可用于食品中小肠结肠炎耶尔森氏菌初筛检测。     

Comparison of DNA extraction methods for pathogenic Yersinia enterocolitica detection from meat food by nested PCR

The objective of this work was to compare three different methods of DNA extraction from meat food, and to determine whether these methods removed inhibitors of nested PCR for pathogenic Yersinia enterocolitica detection. The amplification of the yadA gene from the DNA obtained from a pure Y. enterocolitica culture could be carried out with all the protocols. DNA amplification from the food samples was observed with two of the three tested protocols, which gave highly sensitive amplifications (detection limit 1 CFU/ml). These protocols detected a lower limit of 0.6 fg/μl of DNA extracted from Y. enterocolitica pure culture. We concluded that these protocols were able to eliminate satisfactorily the PCR inhibitors present in the foods. The nested PCR tested could be used satisfactorily in the investigation of pathogenic Y. enterocolitica in foods in the presence of a high background of microflora.     

Yersinia enterocolitica in milk and dairy products

Yersinia enterocolitica was first recognized during the 1960's as an important human enteropathogen. The species as later redefined includes both pathogenic and nonpathogenic forms. Pathogenic strains that retain the virulence plasmid can be identified in several animal models and four indirect tests (calcium dependency, autoagglutination, Congo red uptake, serological detection of outer membrane antigen) and by tissue culture assay, serotype, and biotype. Y. enterocolitica and related bacteria have frequently been isolated from raw milk, but none of the isolates, with the possible exception of serotype 05,27, are recognizable as pathogens. Under normal circumstances Y. enterocolitica does not survive pasteurization. If introduced into pasteurized milk, it can grow well at refrigeration temperatures. Two outbreaks of yersiniosis have occurred that involved pasteurized milk. Pigs, which frequently carry pathogenic Y. enterocolitica in their throat, were the probable source in one of these outbreaks. The most rapid enrichment procedure available for isolation of Y. enterocolitica requires 6 d. No isolation method is available for selective isolation of pathogenic Y. enterocolitica in the presence of related bacteria common in milk and other foods.     

Prevalence of Salmonella in two Botswana abattoir environments

A 1-year study was carried out to investigate the prevalence of Salmonella in two abattoir environments coded "A" and "B" in Gaborone, Botswana. The total number of environmental samples collected from abattoirs A and B was 250 and 300, respectively. The samples were taken from soils in the corrals, knife blades, saw blades, cattle-drinking water, cattle feces, and feed. Preenrichment, enrichment, and selective/differential media, which enabled the favorable growth of Salmonella, were used in the study. Salmonellae were present in all sampled environments. The most common serotypes found in the environment at abattoir A were E1, C1, C2, and B. Serotypes B, C1, C2, C3, and E1 were common in abattoir B. Antigenic characterization of the salmonellae isolates showed that Salmonella Anatum, Salmonella Azteca, Salmonella Saintpaul, Salmonella Cerro, and Salmonella Westhampton were predominant in abattoir A, whereas Salmonella Anatum, Salmonella Mbandaka, Salmonella Molade, Salmonella Reading, and Salmonella Oranienburg were dominant in abattoir B. Implementing hazard analysis critical control point principles in work procedures would definitely reduce the gross contamination taking place in abattoirs.     

Aspects of the epidemiology of Yersinia enterocolitica: a review

A review of works concerning different aspects of the epidemiology of human pathogenic Yersinia enterocolitica biogroup IV/serogroup O:3 (Y. enterocolitica O:3) is given. To investigate the epidemiology of Y. enterocolitica O:3 in Danish herds of pigs, tonsil swabs from 2218 freshly slaughtered pigs originating from 99 herds were examined. The organism was isolated from 25% of the pigs and from 82% of the herds. No herd management factor could be associated with the presence of the organism. The effect of slaughtering technique on surface contamination with Y. enterocolitica O:3 was investigated. 1256 pigs were slaughtered by different evisceration techniques. When a mechanical bung cutter was used instead of the traditional, manual evisceration the contamination was reduced markedly, especially when the rectum and anus were enclosed in a plastic bag prior to the removal of the gut. It was possible to reduce the rate of the surface contamination from 26% on the medial hind limb and 13% on the split sternum and surroundings to about 2% for both sampling sites. An investigation of the presence of Y. enterocolitica O:3 in meat and meat products in retail butcher's shops was performed. The organism was detected in 10 of 33 samples of minced pork and in three of 24 samples of minced beef, but in none of 32 samples of sliced vacuum packed, low to medium salt meat products. The positive minced beef samples were collected at butcher's shops from which positive samples of minced pork were found as well. It is concluded that Y. enterocolitica O:3 is common in pork with a risk of cross-contamination to other products for example ready-to-eat meat products that might be a source of human infection.     

Isolation of Yersinia enterocolitica from foods.

Many selective enrichment and plating media for the isolation of Yersinia enterocolitica from foods are described. However, at present no single isolation procedure is available for the recovery of all pathogenic strains of Yersinia enterocolitica. Cold enrichment in phosphate-buffered saline plus 1% sorbitol and 0.15% bile salts (PBSSB) and two-step enrichment with tryptone soy broth (TSB) and bile oxalate sorbose (BOS) broth are very efficient methods for the recovery of a wide spectrum of serotypes of Y. enterocolitica. Enrichment in irgasan ticarcillin chlorate (ITC) broth was found to be the most efficient method for the recovery of strains of serotype 0:3, which is the most common clinical serotype of Y. enterocolitica in Europe. Post-enrichment alkali treatment often results in higher isolation rates. Cefsulodin irgasan novobiocin (CIN) agar and Salmonella-Shigella deoxycholate calcium chloride (SSDC) agar are the most commonly used plating media. For the recovery of serotype 0:8 strains, the common clinical isolates in North America, enrichment in BOS and plating on CIN seems the most efficient procedure. Selection of the proper enrichment procedure will depend on the bio/serotypes of Yersinia spp. sought and on the type of food to be examined. The use of more than one medium for both enrichment and plating will result in higher recovery rates of Yersinia spp. from foods. Parallel use of the following two isolation procedures is recommended. (1) Enrichment in ITC for 2 days at 24 degrees C; plating on SSDC agar (2 days at 30 degrees C). (2) Pre-enrichment in TSB for 1 day at 24 degrees C; enrichment in BOS for 5 days at 24 degrees C; alkali treatment (mixing 0.5 ml enriched broth with 4.5 ml of 0.5% KOH in 0.5% NaCl for 5 s); plating on CIN agar (2 days at 24 degrees C).     

Inactivation kinetics of Yersinia enterocolitica by citric and lactic acid at different temperatures

Inactivation of Yersinia enterocolitica by citric (1--20% w/v) and lactic (0.3--4.0% v/v) acids at different temperatures (4, 20, 40 degrees C) has been investigated. Inactivation effect of citric and lactic acids was dependent on time and temperature of exposure and acid concentration. Survival curves of Y. enterocolitica suspended in citric acid solutions at 4 and 20 degrees C displayed a shoulder followed by an exponential inactivation, but at 40 degrees C a shoulder was not observed. At all temperatures investigated, survival curves of Y. enterocolitica suspended in lactic acid solutions were linear or slightly concave upwards. A mathematical model based on the Weibull distribution accurately described the kinetics of inactivation of Y. enterocolitica by both acids. The influence of the citric acid concentration on Y. enterocolitica resistance was independent of the treatment temperature. However for lactic acid, the influence of the acid concentration on microbial inactivation depended on the temperature. At any temperature investigated, lactic acid was significantly more effective than citric acid.