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.     

Modelling inactivation of Staphylococcus aureus and Yersinia enterocolitica by high-pressure homogenisation at different temperatures

A detailed study of the inactivation of Staphylococcus aureus and Yersinia enterocolitica by high-pressure homogenisation was performed at, respectively, 25 and 35 different combinations of process temperature and process pressure covering a range of 5-50 degrees C and 100-300 MPa. It appeared that in the entire studied pressure-temperature domain, S. aureus was more resistant to high-pressure homogenisation than Y. enterocolitica. Furthermore, the effect of the process pressure on the inactivation of S. aureus was considerably smaller than on the inactivation of Y. enterocolitica. Also, temperature between 5 and 40 degrees C did not affect inactivation of S. aureus by high-pressure homogenisation, while Y. enterocolitica inactivation was affected by temperature over a much wider range. Different mathematical models were compared to describe the inactivation of both bacteria under the experimental conditions applied. Such pressure-temperature inactivation models form the engineering basis for design, evaluation and optimisation of high-pressure homogenisation processes as a new preservation technique.     

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.     

Predicting thermal inactivation in media of different pH of Salmonella grown at different temperatures

The influence of the growth temperature and the pH of the heating medium on the heat resistance at different temperatures of Salmonella typhimurium ATCC 13311 was studied and described mathematically. The shift of the growth temperature from 10 to 37 degrees C increased heat resistance of S. typhimurium fourfold. The pH of the heating medium at which heat resistance was maximum was pH 6 for cells grown at 37 degrees C, but changed with growth temperature. The alkalinization of the heating medium from pH 6 to pH 7.7 decreased the heat resistance of cells grown at 37 degrees C by a factor of 3. Neither the growth temperature nor the pH modified the z values significantly (4.9 degrees C). The decimal reduction times at different treatment temperatures, in buffers of different pH of cells of S. typhimurium grown at different temperatures, were accurately described by a mathematical equation (correlation coefficient of 0.97). This equation was also tested for Salmonella senftenberg 775W (ATCC 43845) and Salmonella enteritidis ATCC 13076, strains in which the correlation coefficients between the observed and the theoretically calculated values were 0.91 and 0.98, respectively.     

Development and validation of growth model for Yersinia enterocolitica in cooked chicken meats packaged under various atmosphere packaging and stored at different temperatures.

Mathematical models that can predict the growth of Yersinia enterocolitica in chicken meats were evaluated in this study. The growth curves for Y. enterocolitica in chicken meats variously packaged (air, vacuum, and modified atmosphere packaging [MAP]) and stored at various temperatures (4, 10, 16, 22, 28, and 34 degrees C) were constructed. The Gompertz model was applied to fit each of the experimental curves for the conditions mentioned above. The variations in the parameters, including lag time (lambda) and specific growth rate (mu), at various temperatures were then described by the following models: the variations in lag time were described by the Adair and Smith models and the variations in the specific growth rate were described by the Ratkowsky and Zwietering models. The various models were then compared using graphical and mathematical analyses such as mean square error (MSE), regression coefficient (r2), bias factor, and accuracy factor. The results indicate that the mean r values in the Gompertz model for chicken meats packaged in air, vacuum, and MAP were 0.99, 0.99, and 0.95, respectively. The lag time modeled with the Adair and Smith functions exhibited a greater variance and demonstrated larger errors. The MSEs were 0.0015 and 0.0017 for Ratkowsky and Zwietering models, respectively. The r2 values in the Ratkowsky and Zwietering models were both 0.99. The bias factor was 1.017 for the Ratkowsky model and 1.096 for the Zwietering model. The accuracy factor of the Zwietering model was 1.174, which was lower than that in the Ratkowsky model (1.275), indicating that the former model was more accurate than the latter in predicting the specific growth rate of Y. enterocolitica in chicken meats.     

Enrichment procedures and plating media for isolation of Yersinia enterocolitica

A shortened enrichment procedure (25 degrees C for 24 h) was compared with cold enrichment procedures (4 degrees C for 1 to 3 weeks) and direct plating for isolation of Yersinia enterocolitica from commercial ground meat samples. The combined data of all recovery procedures showed that this organism was isolated from 34% of the ground beef samples. The highest isolation rate was 32% for the 4 degrees C/3-week enrichment, followed by 28% for the 4 degrees C/2-week enrichment, 26% for the 25 degrees C/24-h enrichment, 22% for the 4 degrees C/1-week enrichment, and 10% for direct plating. No significant differences (P > 0.05) in isolation rate occurred between the 4 degrees C/3-week, 4 degrees C/2-week, 25 degrees C/24-h, and 4 degrees C/1-week enrichments. The combined data of all recovery procedures showed that Y. enterocolitica was isolated from 64% of ground pork samples. The highest isolation rate was 48% for the 4 degrees C/3-week enrichment, followed by 40% for the 25 degrees C/24-h enrichment, 34% for the 4 degrees C/2-week enrichment, 24% for the 4 degrees C/1-week enrichment, and 24% for direct plating. No significant differences (P > 0.05) in isolation rate occurred between the 4 degrees C/3-week, 25 degrees C/24-h, and 4 degrees C/2-week enrichments. During the plating phase of the experiment, the efficiency of a dye-containing, Yersinia-selective medium (KV202) was compared with that of a commercially available cefsulodin-irgasan-novobiocin medium. Recovery rates were similar for both media. However, KV202 agar differentiated Y. enterocolitica from such contaminating bacteria as Enterobacter, Serratia, and Salmonella by colony morphologic characteristics and color.     

Radiation resistance of virulence plasmid-containing and plasmid-less Yersinia enterocolitica

Yersinia enterocolitica, a foodborne pathogen, can be eliminated from meat by ionizing radiation. Y. enterocolitica sometimes contains a 70-kb virulence plasmid that encodes genes for a type III secretion channel and host immune suppression factors. The radiation resistance of virulence plasmid-containing and plasmid-less Y. enterocolitica was determined. Four Y. enterocolitica serotypes containing (i) the large virulence plasmid, and (ii) their plasmid-less derivatives were inoculated into raw ground pork, which was then vacuum packed and irradiated at 4 degrees C to doses of 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. The D10-value, the radiation dose required to reduce the number of viable Y. enterocolitica by 90%, was not dependent on the presence or absence of the virulence plasmid, but it did differ among the four Y. enterocolitica serotypes.     

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.     

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).     

CpxRA双组分系统对小肠结肠炎耶尔森菌环境耐受性和抗生素耐药性的影响

目的 探究CpxRA双组分系统对小肠结肠炎耶尔森菌环境耐受性和抗生素耐药性的影响。方法 本研究选用Cat-SacB无标记基因敲除系统,利用同源重组双交换原理,构建了基因敲除菌株ΔcpxA和ΔcpxR,以及相应的回补菌株ΔcpxA-cpxA和ΔcpxR-cpxR,然后分别测定了野生型、突变型和回补型在酸性、碱性、高氧和高渗环境中的存活率,以及红霉素、罗红霉素、卡那霉素、链霉素的MIC值。结果 cpxA和cpxR基因的缺失均降低了小肠结肠炎耶尔森菌在酸性、碱性、高氧和高渗环境环境中的存活率;cpxA基因的缺失增强了该菌对于氨基糖苷类抗生素：卡那霉素、链霉素的耐药性,而cpxR基因的缺失减弱了该菌对于大环内酯类抗生素：红霉素、罗红霉素的耐药性。结论 CpxRA双组分系统参与调控小肠结肠炎耶尔森菌在不良环境中的存活能力以及抗生素耐药性。     

Effect of temperature on the radiation resistance of virulent Yersinia enterocolitica

Yersinia enterocolitica, a food-borne pathogen, can be eliminated from meat using ionizing radiation. Commercial facilities may irradiate meat at refrigeration or frozen temperature, or packed in dry ice if the facility does not have refrigeration capabilities. The effect of temperature on the radiation resistance of Y. enterocolitica that contained the 70 kb large virulence plasmid was determined. A mixture of four Y. enterocolitica strains was inoculated into ground pork, which was then vacuum-packed, equilibrated to refrigeration or sub-freezing temperatures, and irradiated to doses of 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. The D(10) value, the radiation dose required to reduce the number of viable Y. enterocolitica by 90%, increased as product temperature decreased with values of 0.19, 0.19, 0.21, 0.40, 0.40. 0.38, and 0.55 kGy being obtained at +5, 0, -5, -10. -15, -20 and -76?°C, respectively. Meat product temperature should be considered when selecting a radiation dose required for elimination of Y. enterocolitica.     

Drug resistance and lecithinase activity of Yersinia enterocolitica isolated from buffalo milk

Two-hundred-and-seven samples of raw buffalo milk and 60 samples of pasteurized buffalo milk were screened for presence of Yersinia enterocolitica. The prevalence of Y. enterocolitica was found to be 24.1% in raw milk, however, no isolation could be made from the pasteurized milk samples. Cold enrichment in trypticase soy broth and alkali treatment methods were followed in this study. The majority of the isolates (62%) were found sensitive to all the antibiotics used and only a few (16%) were resistant to two or more than two antibiotics. The incidence of Y. enterocolitica showed seasonal variations. Incidence was much higher (25-50%) during the winter season as compared to the summer (0-17%). The incidence of lecithinase production was high (40-50%) in Yersinia isolates resistant to one or two antibiotics.     

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.     

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.     

Inactivation of Yersinia enterocolitica by nitrite and nitrate in food.

The antimicrobial effects of sodium nitrite and sodium and potassium nitrate against Yersinia enterocolitica were investigated in solution and in treated pork meat. Potassium nitrate and sodium nitrate showed only feeble antimicrobial activity in cultures; no antimicrobial activity was detected with sodium nitrite. Conversely, all three salts displayed apparent antimicrobial activity in pork meat, possibly due to selective effects on competitive flora.     

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.     

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.     

Occurrence of Yersinia enterocolitica in milk and dairy products in Morocco.

A total of 227 samples of milk and dairy products were examined for the presence of Yersinia enterocolitica. Yersinia spp. were recovered from 11 of 30 raw milks (36.6%), one of 20 pasteurized milks (5%), 15 of 63 traditional fermented milks (23.8%), seven of 94 cheeses and one of 20 cream samples (5%). The overall incidence of Y. enterocolitica in milk and dairy products was 6.6%. The other Yersinia species were Y. intermedia, Y. kristensenii, Y. frederiksenii and Y. pseudotuberculosis. Y. enterocolitica was detected only in raw milk (30% of the samples), in traditional fermented milks (6.3%) and in raw milk-made cheese (4%). The majority of the Y. enterocolitica isolates were of biotype 1 (environmental strains). The Celfulodin-Irgasan-Novobiocin (CIN) Agar was found to be more efficient than the Mac Conkey Agar in the isolation of Yersinia organisms.