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 and characterization of pathogenic Yersinia enterocolitica in pig tonsils from different slaughterhouses

The prevalence of yadA-positive Yersinia enterocolitica was determined in 185 pig tonsils from nine slaughterhouses using both the PCR and culture method. The mean prevalence was 37%, varying from 13% to 45% when both PCR and culture-positive results were included. Of the 52 PCR-positive tonsil samples, 20 were culture-negative, while of the 48 culture-positive, 16 were PCR-negative. Using the culture method, Y. enterocolitica belonging to the bioserotype 4/O:3 was found in 61 tonsils, of which 48 were yadA-positive. Type 4/O:3 was the only pathogenic bioserotype found in this study. Most of the yadA-positive samples (85%) were recovered already after overnight enrichment. A total of 61 isolates, including 13 yadA-negative isolates from different samples, were characterized with PFGE. UsingNotI and XbaI, 20 and 17 PFGE patterns were obtained, respectively. Although the patterns were not identical, most of them played only minor deviations. A total of 26 pulsotypes, defined by combination of the various NotI andXbaI digestion profiles, were observed. Two to eight different pulsotypes were observed in each slaughterhouse, The most common pulsotypes, 1a and 4g, were found in 36% and 20% of the tonsils, respectively and these pulsotypes were widely distributed to most of the slaughterhouses. The pulsotype 1a was identified in eight out of nine slaughterhouses and the pulsotype 4g in seven slaughterhouses.     

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.     

Growth and survival of Yersinia enterocolitica at acidic pH

The ability of three strains of Yersinia enterocolitica to survive and grow in tryptic soy broth (TSB) at pH 3.0, 3.5, 4.0, 4.5, 5.0, 5.5 and 6.0 was determined. In addition, experiments were done to assess survival of Y. enterocolitica in mayonnaise, spoonable salad dressing, and tartar sauce. Two of the strains were able to grow in TSB at pH 4.5 and above when incubated at 25°C. Cell concentrations of all strains decreased at lower pH values at this temperature. No growth of Yersinia was observed when TSB was incubated at 5°C for any pH tested, although survival was prolonged. Viable cells were not recovered from artificially contaminated tartar sauce (pH 3.2) or spoonable salad dressing (pH 3.4) at any time. Cells survived for at least 48 h in artificially contaminated mayonnaise (pH 3.8) held at 5°C. No viable cells were recovered after 24 h from mayonnaise held at 21°C.     

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.     

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.     

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.     

自腹泻病例和家用冰箱分离的小肠结肠炎耶尔森菌病原特征分析

目的研究自腹泻病例和家用冰箱中分离的小肠结肠炎耶尔森菌(Yersinia enterocolitica,Ye)的病原学特征,为科学防控Ye的污染提供依据。方法采集2017年北京市顺义区2家哨点医院肠道门诊腹泻患者病例粪便标本和顺义区83户家用冰箱中的涂抹拭子样品,对分离的Ye进行毒力基因、脉冲场凝胶电泳(PFGE)分子分型检测和耐药试验。结果腹泻病例粪便标本中Ye阳性率为0.27%(1/372),家用冰箱中Ye阳性率为6.02%(5/83);冰箱分离株仅携带ystB基因,腹泻病例分离株携带ail、ystA、virF、yadA基因;腹泻病例和冰箱分离株PFGE带型亲缘关系较远。全部Ye对氨苄西林、阿莫西林/克拉维酸、头孢唑林均耐药,腹泻病例分离株对萘啶酸耐药。结论 Ye在北京市顺义区腹泻病例中流行强度不高,家用冰箱受Ye污染较为严重,腹泻病例和冰箱中分离的Ye病原学特征具有一定差异。     

Stability of the virulence plasmid in Yersinia enterocolitica at elevated temperatures

This report presents information about the stability of the Yersinia enterocolitica virulence plasmid at elevated temperatures. Though considerable variation existed in the heat resistance of five strains (representing four serotypes), exposure of the different plasmid-bearing virulent strains to temperatures of 45, 50 and 55°C did not cause the loss of the virulence plasmid from the surviving cells. Based on the crystal violet (CV) binding test, these surviving cells were still virulent.     

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.     

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.     

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.     

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。