Yersinia enterocolitica in slaughter pig tonsils: Enumeration and detection by enrichment versus direct plating culture

Tonsil samples from 139 slaughter pigs were examined for the presence of pathogenic Yersinia enterocolitica by enrichment procedures based on the standard method ISO 10273:2003. In addition, samples were tested by direct plating method to evaluate its efficiency compared to the enrichment culture methods and to quantify the level of contamination in porcine tonsils. In total, 52 samples (37.4%) were positive for pathogenic Y. enterocolitica, all belonging to bioserotype 4/O:3. Fifty out of the 52 positive samples (96.2%) were detected by direct plating. Enumeration showed an average concentration of 4.5 log₁₀ CFU g⁻¹ and 4.4 log₁₀ CFU g⁻¹ tonsil on Salmonella-Shigella-desoxycholate-calcium chloride (SSDC) and cefsulodin-irgasan-novobiocin (CIN) agar plates, respectively. The enrichment procedures recommended by the ISO 10273:2003 method were not optimal for the isolation of pathogenic Y. enterocolitica from pig tonsils: two days enrichment in irgasan-ticarcillin-potassium chlorate (ITC) broth resulted in an isolation rate of 84.6%, while 5 days enrichment in peptone-sorbitol-bile (PSB) broth recovered only 59.6% of positive samples. Reducing the enrichment time in PSB from 5 to 2 days resulted in a significantly higher recovery rate (94.2%) and might serve as an appropriate enrichment protocol for the isolation of pathogenic Y. enterocolitica from pig tonsils. Compared to enrichment culture methods, results based on direct plating can be obtained in a shorter time course and provide quantitative data that might be needed for further risk assessment studies.     

Thermal inactivation of Yersinia enterocolitica in pork slaughter plant scald tank water

The objective of this study was to establish the time–temperature combinations required to ensure the thermal inactivation of Yersinia enterocolitica during scalding of pork carcasses. A 2 strain cocktail of Y. enterocolitica (bioserotypes 2/O:5,27 and 1A/O:6,30) was heat treated at 50, 55 and 60 °C in samples of scald tank water obtained from a commercial pork slaughter plant. Samples were removed at regular intervals and surviving cells enumerated using (i) Cefsulodin–Irgasan–Novobiocin Agar (CIN) supplemented with ampicillin and arabinose and (ii) Tryptone Soya Agar (TSA), overlaid with CIN agar with ampicillin and arabinose. The data generated was used to estimate D- and z-values and the formula D_x = log^− 1(log D₆₀ − ((t₂ − t₁)/z)) was applied to calculate thermal death time–temperature combinations from 55 to 65 °C. D₅₀, D₅₅ and D₆₀-values of 45.9, 10.6 and 2.7 min were calculated from the cell counts obtained on CIN agar, respectively. The corresponding D-values calculated from the TSA/CIN counts were 45.1, 11 and 2.5 min, respectively. The z-value was 7.8. It was concluded that a time–temperature combination of 2.7 min at 60 °C is required to achieve a 1 log reduction in Y. enterocolitica in pork scald tank water. The predicted equivalent at 65 °C was 0.6 min. This study provides data and a model to enable pork processors to identify and apply parameters to limit the risk of carcass cross-contamination with Y. enterocolitica in pork carcass scald tanks.     

Detection and prevalence of pathogenic Yersinia enterocolitica in refrigerated and frozen dairy products by duplex PCR and dot hybridization targeting the virF and ail genes

Pathogenic Yersinia enterocolitica is involved in yersiniosis through expression of chromosome-borne or plasmid-borne virulence factors. Yersinia enterocolitica is a cold-tolerant pathogen frequently isolated from refrigerated or frozen foods. However, little attention has been focused on the prevalence of pathogenic Y. enterocolitica in refrigerated or frozen dairy samples in China. In this study, we developed a new duplex PCR targeting the plasmid-borne virF gene and chromosome-borne ail gene for detection of pathogenic Y. enterocolitica isolates. We established a detection limit for the duplex PCR of 6.5 × 10² cfu/mL in artificially contaminated dairy samples. In addition, the duplex PCR could detect directly 4.5 to 5.7 cfu of Y. enterocolitica in 5 mL of brain heart infusion broth after 6 h of enrichment at 28°C. A newly developed dot hybridization assay further confirmed specificity of the duplex PCR for detection of virulent Y. enterocolitica. Furthermore, 13 Y. enterocolitica and 5 pathogenic strains, from 88 commercial frozen or refrigerated dairy products, were detected successfully by the China National Standard method (GB/T4789.8-2008) and the duplex PCR, respectively. Finally, biotypes and serotypes of pathogenic Y. enterocolitica strains were further characterized. The duplex PCR developed here is reliable for large-scale screening, routine monitoring, and risk assessment of pathogenic Y. enterocolitica in refrigerated or frozen dairy products.     

Prevalence of pathogenic Yersinia enterocolitica and Yersinia pseudotuberculosis in wild boars in Switzerland

Between October 2007 and March 2008, 153 wild boars shot in the Canton of Geneva in Switzerland were sampled. Fifty-one percent of the animals were males and 49% were females. The age of most (81%) animals varied between 6 months and 2 years. Prevalence of enteropathogenic Yersinia in tonsils and faeces was studied using culture and PCR methods and in tissue fluid of tonsils using an ELISA system. Prevalence of anti-Yersinia antibodies in tissue fluid was 65%. Detection rate of enteropathogenic Yersinia in tonsils of 153 wild boars by real-time PCR was 44%. Ail-positive Yersinia enterocolitica and inv-positive Yersinia pseudotuberculosis were detected in 35 and 20% of the animals, respectively. Both species were detected in 10% of the animals. Isolation rate of enteropathogenic Yersinia was low; ail-positive Y. enterocolitica and inv-positive Y. pseudotuberculosis were found in 9 and 3% of the animals, respectively. Prevalence was shown to be significantly higher in tonsils than in faeces. Furthermore, females were more commonly positive than males. This study shows that the prevalence of enteropathogenic Yersinia is high and both enteropathogenic Y. enterocolitica and Y. pseudotuberculosis are common findings in tonsils of wild boars in Switzerland.     

Pathogenic Yersinia enterocolitica 2/O:9 and Yersinia pseudotuberculosis 1/O:1 strains isolated from human and non-human sources in the Plateau State of Nigeria

Foodborne yersiniosis, caused by enteropathogenic Yersinia, especially Yersinia enterocolitica, is an important cause of diarrhea in developed countries, especially in temperate zones. Since studies concerning the presence of enteropathogenic Yersinia in humans and foods are rare in developing countries and tropical areas, human and non-human samples were studied in Plateau state of Nigeria to obtain information on the epidemiology of Y. enterocolitica and Yersinia pseudotuberculosis. Surprisingly, ail-positive Y. enterocolitica and inv-positive Y. pseudotuberculosis were isolated in Plateau state of Nigeria from several samples of human and non-human origin. Bioserotype 1/O:1 was the only Y. pseudotuberculosis type found. Y. enterocolitica belonging to bioserotype 2/O:9 was the dominating type found in most samples. Bioserotype 4/O:3 was isolated only from one pig and one sheep. Using PFGE, 5 genotypes were obtained among 45 Y. enterocolitica 2/O:9 strains with NotI, ApaI and XhoI enzymes and 3 among 20 Y. pseudotuberculosis 1/O:1 strains with NotI and SpeI enzymes. All human Y. pseudotuberculosis 1/O:1 strains were indistinguishable from pig, sheep or food strains. The dominating genotype of Y. enterocolitica 2/O:9 strains among humans was also found among strains isolated from pig, fermented cow milk and traditional intestine pepper soap samples.     

Comparison of Yersinia pestis to other closely related Yersinia species using fatty acid profiles

Capillary gas chromatography with flame ionisation detection (GC–FID) was used to determine the cellular fatty acid (CFA) profiles of six Yersinia pestis strains. The profiles were then compared with the CFA profiles of other closely related Yersinia species including: Y. pseudotuberculosis, Y. enterocolitica, Y. intermedii, Y. kristensenii and Y. frederiksenii. For GC–FID analysis, whole cell fatty acid methyl esters (FAMEs) from cells cultured on brain–heart infusion (BHI) agar at 35 °C for 24 h were obtained by saponification, methylation and extraction into hexane/methyl tert-butyl ether. A data set for each Yersinia species was prepared using fatty acid profiles from five replicates prepared on different days. Major fatty acids of the 26 Yersinia strains evaluated in this study were straight-chain 12:0, 14:0, 15:0, 16:0 and unsaturated summed 16:1 ω7c/16:1 ω6c, 18:1 ω7c, and summed 14:0 3OH/16:1 iso, and 17:0 ω cyclo 7–8. The CFA profiles for Y. pestis and Y. pseudotuberculosis are similar, but there are several fatty acids, 16:1 ω5c, 16:0, 17:1 ω7c, 17:0 ω cyclo 7–8, 19:0 and summed 18:2 ω6c, 9c/18:0 ante, that differ significantly between these two species. Analysis of FAMEs from Yersinia strains grown on BHI agar by a rapid GC–FID method can provide a sensitive procedure for the identification of these organisms, and this analytical method provides a procedure for the differentiation of Y. pestis strains from closely related Yersinia species.     

温州市食品中小肠结肠炎耶尔森氏菌分布及分子分型研究

目的 了解温州市食品中小肠结肠炎耶尔森氏菌的分子分型及分布特征。方法 4 ℃增菌后用选择性培养基对食品中的小肠结肠炎耶尔森氏菌进行分离鉴定,分析阳性菌株的生物型、血清型、毒力基因型、耐药性和脉冲场凝胶电泳（PFGE）分子型别。结果 采集6类食品,共676份样品,其中68份样品检出69株小肠结肠炎耶尔森氏菌,检出率为10.1%（68/676）。调理肉制品检出率最高（20.5%,9/44）,其次为速冻食品（17.2%,11/64）。95.7%（66/69）的分离株为生物1A型,生物血清型以1A/O∶5（29.0%）为主,其次为1A/O∶8（14.5%）;88.4%（61/69）的菌株仅携带ystB基因,检出1株4/O∶3型菌株携带毒力基因ail、ystA、yadA、virF。分离株对14种抗菌药物的敏感率达到94%以上。32株血清已分型菌株,分为29种PFGE带型。结论 温州市食品中存在一定程度的小肠结肠炎耶尔森氏菌污染,且检出致病性小肠结肠炎耶尔森菌,菌株耐药率处于较低水平,分子分型提示菌株呈高度遗传多态性。     

Multiplex fiber optic biosensor for detection of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella enterica from ready-to-eat meat samples

Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella enterica are the most common foodborne bacterial pathogens and are responsible for many outbreaks. Therefore, multiplex detection of these three using a single assay platform is highly desirable. The objective was to develop and optimize a fiber optic sensor for simultaneous detection of these three from food. The streptavidin coated optical waveguides were immobilized with biotinylated polyclonal antibodies and exposed to the bacterial suspensions or enriched food samples for 2 h. Pathogens were detected after reacting with Alexa-Fluor 647-labeled monoclonal antibodies. Ready-to-eat beef, chicken and turkey meats were inoculated with each pathogen (∼100 cfu/25 g), enriched in SEL (Salmonella, E. coli, Listeria), a multipathogen selective enrichment broth for 18 h and tested with the biosensor. The biosensor was able to detect each pathogen, individually or in a mixture with very little cross-reactivity. The limit of detection for the sensor was ∼10³ cfu/ml for all three pathogens. Furthermore, the biosensor successfully detected each pathogen, grown in a mixture from enriched meat samples under 24 h. The pathogen presence was further verified by PCR and immunofluorescence assay. The multiplex fiber optic sensor shows promise for detection of the three pathogens if present in the same sample eliminating the use of multiple single pathogen detection platforms.     

Satureja horvatii essential oil: In vitro antimicrobial and antiradical properties and in situ control of Listeria monocytogenes in pork meat

The dominant compounds in Satureja horvatii oil were p-cymene (33.14%), thymol (26.11%) and thymol methyl ether (15.08%). The minimum inhibitory concentration (MIC) varied from 0.03 to 0.57 mg/mL for bacteria, and from 0.56 to 2.23 mg/mL for yeast strains, while minimum bactericidal/yeast-cidal concentration (MBC/MYC) varied from 0.07 to 1.15 mg/mL and 1.11 to 5.57 mg/mL for bacteria and yeasts, respectively. The antiradical potential of the essential oil was evaluated using hydroxyl radical (•OH) generated in Fenton reaction. The meat preserving potential of essential oil from Satureja horvatii was investigated against L. monocytogenes. Essential oil successfully inhibited development of L. monocytogenes in pork meat. Sensorial evaluation on flavor and color of meat was performed. The color and flavor of meat treated with essential oil improved after 4 days of storage. S. horvatii essential oil can act as a potent inhibitor of food spoiling microorganisms, in meat products and also can be a useful source of natural antioxidants.     

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

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

Comparison of culture media for enrichment and isolation of Salmonella spp. from frozen Channel catfish and Vietnamese basa fillets

Frozen fillets of Channel catfish and Vietnamese basa fish were used to compare Salmonella spp. recovery effectiveness of selective enrichment in Rappaport–Vassiliadis (RV) broth and tetrathionate broth (TT) and selective isolation on Hekteon enteric (HE) agar, xylose lysine deoxycholate (XLD) agar, and bismuth sulfite (BS) agar. Isolate confirmation was through fatty acid methyl ester analysis. Of 60 samples analyzed, 25 were found contaminated with Salmonella (42% incidence). Salmonella spp. recovery after enrichment in RV medium was 35% on HE agar, 30% on XLD agar, and 42% on BS agar. Similarly, after enrichment in TT broth, HE and XLD agars recovered 22% each and BS agar recovered 37%. No performance difference (p > 0.05) was observed in the recovery of Salmonella using the combinations of BS, HE, and XLD agars with RV broth and BS agar with TT broth. The combination of selective enrichment in RV and selective isolation on BS gave numerically greatest isolation of Salmonella from Channel catfish and Vietnamese basa fish compared to other isolation combinations.     

Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat

In this study, a combined enrichment/real-time PCR method for the rapid detection of Salmonella on fresh meat carcasses, was designed, developed and validated in-house following requirements outlined in ISO 16140:2003. The method included an 18 h non-selective enrichment in buffered peptone water (BPW) and a 6 h selective enrichment in Rappaport Vasilliadis Soya (RVS) broth, based on the traditional culture method, ISO 6579:2002. The real-time PCR assay included an internal amplification control (IAC), was 100% specific and was sensitive to one cell equivalent. The alternative method was validated against the traditional culture method and relative accuracy of 94.9%, sensitivity of 94.7% and specificity of 100% were determined using 150 fresh meat carcass swabs. This alternative method had a detection limit of 1–10 CFU/100 cm² for fresh meat carcass swabs and was performed in 26 h. Following further inter-laboratory studies, this alternative method could be suitable for implementation in testing laboratories for the analysis of carcass swabs.     

Application of Classification and Regression Tree (CART) analysis on the microflora of minced meat for classification according to Reg. (EC) 2073/2005

In a retrospective study on the microbiology of minced meat from small food businesses supplying directly to the consumer, the relative contribution of meat supplier, meat species and outlet where meat was minced was assessed by "Classification and Regression Tree" (CART) analysis. Samples (n=888) originated from 129 outlets of a single supermarket chain. Sampling units were 4-5 packs (pork, beef, and mixed pork-beef). Total aerobic counts (TACs) were 5.3±1.0 log CFU/g. In 75.6% of samples, E. coli were <1 log CFU/g. The proportion of "unsatisfactory" sample sets [as defined in Reg. (EC) 2073/2005] were 31.3 and 4.5% for TAC and E. coli, respectively. For classification according to TACs, the outlet where meat was minced and the "meat supplier" were the most important predictors. For E. coli, "outlet" was the most important predictor, but the limit of detection of 1 log CFU/g was not discriminative enough to allow further conclusions.     

Pulsed field, PCR ribotyping and multiplex PCR analysis of Yersinia enterocolitica strains isolated from meat food in San Luis Argentina

The characterization of phenotypic and genotypic virulence markers of Yersinia enterocolitica strains belonging to biotypes (B) 1A, 2 and 3, mostly isolated from food in San Luis, Argentina, and the assessment of their genotypic diversity using PFGE and PCR ribotyping, were performed in our laboratory for the first time. Thirty five Y. enterocolitica strains, two reference strains and 33 strains isolated in our laboratory were studied. The presence of virF, ail, ystA, and myfA genes was investigated by multiplex PCR. The pathogenic potential of B1A strains, the most predominant biotype of Y. enterocolitica strains isolated from meat in our region, was investigated by simple PCR. Four B1A strains were positive for ystB gene. Four Y. enterocolitica 2/O:9 (bio/serotype) and two 3/O:5 strains isolated in our laboratory showed virulence-related results in the phenotypic tests and multiplex PCR. A good correlation between the expression of virulence markers and their corresponding genotypes was observed for most strains. Sixteen genomic types (GT) and 9 different intergenic spacer region (SR) groups were generated by PFGE and PCR ribotyping, respectively. In both cases the Y. enterocolitica 2/O:9 strains were separately clustered from 1A and 3/O:5 strains. Meat foods might be vehicles of transmission of pathogenic Y. enterocolitica strains in our region.     

2022年上海市生禽肉和调理肉制品中主要食源性致病菌监测

目的 分析上海市市售生禽肉和调理肉制品中沙门菌、单核细胞增生李斯特菌及小肠结肠炎耶尔森氏菌等食源性致病菌的污染情况。方法 2022年1～8月,从上海市农贸市场、超市、餐饮店等环节采集样品348份,其中生禽肉240份,调理肉制品108份。按照食品安全国家标准分别对样品中的沙门菌、单核细胞增生李斯特菌和小肠结肠炎耶尔森氏菌的检测。采用VITEK2全自动微生物鉴定仪对可疑菌株进行生化鉴定,并对沙门菌分离株进行血清学分型。结果 生禽肉中沙门菌、单核细胞增生李斯特菌和小肠结肠炎耶尔森氏菌的检出率分别为28.33%(68/240)、5.00%(12/240)和0.83%(2/240);调理肉制品中沙门菌、单核细胞增生李斯特菌和小肠结肠炎耶尔森氏菌的检出率分别为5.56%(6/108)、28.70%(31/108)和1.85%(2/108);血清学分型结果显示,生禽肉中68株沙门菌分布于14种不同血清型,其中科瓦利斯沙门菌（35.29%）、鼠伤寒沙门菌（16.18%）和肠炎沙门菌（13.24%）占比较高,调理肉制品中6株沙门菌分布于3种不同血清型,分别为肠炎沙门菌（66.67%）、肯塔基沙门菌（16...     

Detection and characterization of Salmonella in lairage, on pig carcasses and intestines in five slaughterhouses

In this study, conducted at five slaughterhouses, individual pigs were sampled and followed up from stunning to cooling down of the carcasses. In this way, Salmonella prevalence and possible risk points were described. At the lairage area, pens were sampled using overshoes. At stunning and bleeding, pigs were individually identified and subsequently swabs were taken of the oral cavity and the carcass after polishing, splitting and forced chilling. Additionally, duodenum, ileum, rectum and mesenteric lymph nodes were extracted and samples were taken of the scalding water. All samples were submitted to Salmonella isolation and Salmonella isolates were serotyped and genotyped by pulsed-field gel electrophoresis (PFGE). Of all samples taken (n = 1953), 14.1% were Salmonella positive. The prevalence of S. in the lairage area varied widely (from 0 to 100%) between the slaughterhouses. Of the sampled pigs (n = 226), 48.2% were positive in at least one sample. Statistical analysis revealed that the contamination of the lairage area was related to a higher amount of positive carcasses after polishing. Furthermore, the contamination of the carcasses after splitting and forced chilling was related to the contamination level of the carcass after polishing. A relation between the outer (carcass) contamination and the inner (gut content and lymph nodes) contamination of a pig could not be established. The predominant serotypes were S. Typhimurium (58.7%) and S. Derby (17.4%). Genotyping revealed 46 different PFGE profiles among the 276 Salmonella isolates. The same genotype at the lairage area as in the oral cavity of the pigs was found in 95%. The results indicate that the lairage area is a primary source of Salmonella in slaughter pigs and that carcass contamination originates from the environment rather than from the pig (inner contamination) itself. It further shows that slaughterhouses vary in their capability of dealing with Salmonella positive pigs. A slaughterhouse specific approach is needed, however, general guidelines should be provided to decrease the contamination level of the lairage area and the slaughter environment.     

Formation and identification of nitrosylmyoglobin by Staphylococcus xylosus in raw meat batters: A potential solution for nitrite substitution in meat products

Staphylococcus xylosus and Pediococcus pentosaceus isolated from Chinese dried sausage were assessed for their ability to convert metmyoglobin into nitrosylmyoglobin in Mann–Rogosa–Sharp broth model systems and raw pork meat batters without the addition of nitrite. The results showed that samples in model systems with S. xylosus cultures had an absorption spectra that is typical of nitrosylmyoglobin, an obvious pink colour (judged by visual inspection) and a significantly higher a*-value than the control samples or samples inoculated with P. pentosaceus. In raw meat batters, the a*-values of the S. xylosus samples were almost the same as those for the meat with nitrite added. The complementary analysis of meat batter samples by photochemical information from UV–vis, electron spin resonance and resonance Raman spectroscopy revealed that the existing status of the myoglobin in meat batters inoculated with S. xylosus was mainly pentacoordinate nitrosylmyoglobin. This study provides a potential solution for nitrite substitute in meat products.     

Quantifying Listeria monocytogenes prevalence and concentration in minced pork meat and estimating performance of three culture media from presence/absence microbiological testing using a deterministic and stochastic approach

Listeria monocytogenes poses a serious threat to public health, and the majority of cases of human listeriosis are associated with contaminated food. Reliable microbiological testing is needed for effective pathogen control by food industry and competent authorities. The aims of this work were to estimate the prevalence and concentration of L. monocytogenes in minced pork meat by the application of a Bayesian modeling approach, and also to determine the performance of three culture media commonly used for detecting L. monocytogenes in foods from a deterministic and stochastic perspective. Samples (n = 100) collected from local markets were tested for L. monocytogenes using in parallel the PALCAM, ALOA and RAPID'L.mono selective media according to ISO 11290-1:1996 and 11290-2:1998 methods. Presence of the pathogen was confirmed by conducting biochemical and molecular tests. Independent experiments (n = 10) for model validation purposes were performed. Performance attributes were calculated from the presence–absence microbiological test results by combining the results obtained from the culture media and confirmative tests. Dirichlet distribution, the multivariate expression of a Beta distribution, was used to analyze the performance data from a stochastic perspective. No L. monocytogenes was enumerated by direct-plating (<10 CFU/g), though the pathogen was detected in 22% of the samples. L. monocytogenes concentration was estimated at 14–17 CFU/kg. Validation showed good agreement between observed and predicted prevalence (error = −2.17%). The results showed that all media were best at ruling in L. monocytogenes presence than ruling it out. Sensitivity and specificity varied depending on the culture-dependent method. None of the culture media was perfect in detecting L. monocytogenes in minced pork meat alone. The use of at least two culture media in parallel enhanced the efficiency of L. monocytogenes detection. Bayesian modeling may reduce the time needed to draw conclusions regarding L. monocytogenes presence and the uncertainty of the results obtained. Furthermore, the problem of observing zero counts may be overcome by applying Bayesian analysis, making the determination of a test performance feasible.     

A model for lactic acid-induced inhibition of Yersinia enterocolitica in mono- and coculture with Lactobacillus sakei

Most traditional predictive models for growth of foodborne pathogenic (or spoilage) organisms focus on single species behaviour. This approach may lead to a significant discrepancy between model predictions and reality, since the (potential) influence of the background flora is neglected. In this paper, a specific type of multiple species interaction is considered, namely, a pathogenic organism in the presence of a microbial antagonist, where the latter species inhibits the pathogen's growth through lactic acid production. As an experimental case study, growth curves of Yersinia enterocolitica in mono- and coculture with Lactobacillus sakei were generated in duplicate in a modified brain–heart infusion medium. A complete factorial design was applied to assess the impact of temperature (4 levels) and inoculum ratio pathogen/antagonist (6 levels) on the Y. enterocolitica growth. Based on a set of formulated model requirements, a novel model was developed, in which the experimentally observed inhibition effect, i.e., an (early) induction of the stationary phase in the pathogen's growth curve, is explicitly related to the lactic acid production curve. Experimental data and model predictions show good agreement.     

Urolithins, ellagitannin metabolites produced by colon microbiota, inhibit Quorum Sensing in Yersinia enterocolitica: Phenotypic response and associated molecular changes

The mammalian enteropathogen Yersinia enterocolitica produces two main N-acylhomoserine lactones (AHLs) involved in Quorum Sensing (QS)-mediated infection processes, such as virulence, biofilm maturation and motility. Ellagitannin (ET)-rich fruits exhibit anti-QS activity but in vivo effects against intestinal pathogens may be associated to the ETs gut microbiota derived metabolites, urolithin-A (Uro-A) and urolithin-B (Uro-B). In this work we show that urolithins, at concentrations achievable in the intestine through the diet, reduce the levels of N-hexanoyl-l-homoserine lactone (C6-HSL) and N-(3-oxo-hexanoyl)-l-homoserine lactone (3-oxo-C6-HSL) in Y. enterocolitica and inhibit QS-associated biofilm maturation and swimming motility. These inhibitory effects were not associated to downregulation of the expression of some of the genes involved in the synthesis of AHLs (yenI and yenR) or in motility (flhDC, fliA, fleB). Our results suggest that urolithins may exert antipathogenic effects in the gut against Y. enterocolitica and highlight the need to investigate the antipathogenic in vivo properties of plant derived metabolites.