Widespread listeriosis outbreak attributable to pasteurized cheese, which led to extensive cross-contamination affecting cheese retailers, Quebec, Canada, 2008

A major Listeria monocytogenes outbreak occurred in the province of Quebec, Canada, in 2008, involving a strain of L. monocytogenes (LM P93) characterized by pulsed-field gel electrophoresis (PFGE) and associated with the consumption of pasteurized milk cheese. This report describes the results of the ensuing investigation. All individuals affected with LM P93 across the province were interviewed with a standardized questionnaire. Microbiological and environmental investigations were conducted by the Quebec's Food Inspection Branch of Ministère de l'Agriculture, des Pêcheries et de l'Alimentation du Québec among retailers and cheese plants involved in the outbreak. Between 8 June and 31 December 2008, 38 confirmed cases of LM P93 were reported to public health authorities, including 16 maternal-neonatal cases (14 pregnant women, and two babies born to asymptomatic mothers). The traceback of many brands of cheese that tested positive for LM P93 collected from retailers identified two cheese plants contaminated by L. monocytogenes strains on 3 and 4 September. PFGE profiles became available for both plants on 8 September, and confirmed that a single plant was associated with the outbreak. Products from these two plants were distributed to more than 300 retailers in the province, leading to extensive cross-contamination of retail stock. L. monocytogenes is ubiquitous, and contamination can occur subsequent to heat treatment, which usually precedes cheese production. Contaminated soft-textured cheese is particularly prone to bacterial growth. Ongoing regulatory and industry efforts are needed to decrease the presence of Listeria in foods, including pasteurized products. Retailers should be instructed about the risk of cross-contamination, even with soft pasteurized cheese and apply methods to avoid it.     

Short communication: Nα-Lauroyl-l-arginine ethylester monohydrochloride reduces bacterial growth in pasteurized milk

Effective strategies for extending fluid milk product shelf-life by controlling bacterial growth are of economic interest to the dairy industry. To that end, the effects of addition of l-arginine, Nα-lauroyl ethylester monohydrochloride (LAE) on bacterial numbers in fluid milk products were measured. Specifically, LAE was added (125, 170, or 200 mg/L) to conventionally homogenized and pasteurized 3.25% fat chocolate or unflavored milk products. The treated milks and corresponding untreated controls were held at 6°C and plated on standard plate count agar within 24 h of processing and again at 7, 14, 17, and 21 d of storage. Bacterial counts in all unflavored milk samples treated with LAE remained below the Pasteurized Milk Ordinance limit for grade A pasteurized fluid milk of 4.3 log cfu/mL for the entire 21 d. Bacterial counts in unflavored samples containing 170 and 200 mg/L of LAE were significantly lower than those in the untreated unflavored milk at d 17 and 21 postprocessing. Specifically, bacterial counts in the milk treated with 200 mg/L of LAE were 5.77 log cfu/mL lower than in untreated milk at 21 d postprocessing. Bacterial counts in chocolate milk treated with 200 mg/L of LAE were significantly lower than those in the untreated chocolate milk at d 14, 17, and 21. In chocolate milk treated with 200 mg/L of LAE, bacterial counts were 0.9 log cfu/mL lower than in the untreated milk at 21 d postprocessing. Our results show that addition of LAE to milk can reduce bacterial growth. Addition of LAE is more effective at controlling bacterial growth in unflavored milk than in chocolate milk.     

Development and validation of a dynamic growth model for Listeria monocytogenes in fluid whole milk

Listeria monocytogenes, a psychrotrophic microorganism, has been the cause of several food-borne illness outbreaks, including those traced back to pasteurized fluid milk and milk products. This microorganism is especially important because it can grow at storage temperatures recommended for milk (< or =7 degrees C). Growth of L. monocytogenes in fluid milk depends to a large extent on the varying temperatures it is exposed to in the postpasteurization phase, i.e., during in-plant storage, transportation, and storage at retail stores. Growth data for L. monocytogenes in sterilized whole milk were collected at 4, 6, 8, 10, 15, 20, 25, 30, and 35 degrees C. Specific growth rate and maximum population density were calculated at each temperature using these data. The data for growth rates versus temperature were fitted to the Zwietering square root model. This equation was used to develop a dynamic growth model (i.e., the Baranyi dynamic growth model or BDGM) for L. monocytogenes based on a system of equations which had an intrinsic parameter for simulating the lag phase. Results from validation of the BDGM for a rapidly fluctuating temperature profile showed that although the exponential growth phase of the culture under dynamic temperature conditions was modeled accurately, the lag phase duration was overestimated. For an alpha0 (initial physiological state parameter) value of 0.137, which corresponded to the mean temperature of 15 degrees C, the population densities were underpredicted, although the experimental data fell within the narrow band calculated for extreme values of alpha0. The maximum relative error between the experimental data and the curve based on an average alpha0 value was 10.42%, and the root mean square error was 0.28 log CFU/ml.     

Antimicrobial efficacy of eugenol microemulsions in milk against Listeria monocytogenes and Escherichia coli O157:H7

The antimicrobial activity of eugenol microemulsions (eugenol encapsulated in surfactant micelles) in ultrahigh-temperature pasteurized milk containing different percentages of milk fat (0, 2, and 4%) was investigated. Antimicrobial microemulsions were prepared from a 5% (wt) aqueous surfactant solution (Surfynol 485W) with 0.5% (wt) eugenol. Two strains each of Listeria monocytogenes and Escherichia coli O157:H7 previously shown to be the least and most resistant to the microemulsion in microbiological media were used to inoculate sterile milk (10(4) CFU/ml). Samples were withdrawn and plated at 0, 1, 3, 6, 12, and 24 h for enumeration. Microemulsions completely prevented growth of L. monocytogenes for up to 48 h in skim milk and reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h. Similarly, in 2% fat milk, eugenol-Surfynol combinations reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h but only increased the lag phase of both strains of L. monocytogenes. In full-fat milk (4% fat), microemulsions inhibited growth of the least resistant strains of L. monocytogenes and E. coli but were ineffective against the two resistant strains. Unencapsulated eugenol was slightly more or as inhibitory as microemulsions against target pathogens. Results were attributed to diffusional mass transport of antimicrobials from microemulsions to the macroemulsion (milk). Results suggest that food composition, especially fat level, may affect the efficiency of targeting of foodborne pathogens with surfactant-encapsulated antimicrobials.     

High incidence of Listeria monocytogenes in European red smear cheese

The incidence of Listeria and Listeria monocytogenes in European red smear cheese was determined in order to assess whether the lack of recent outbreaks of listeriosis associated with cheese is due to improved hygenic conditions in the dairies. Out of European red-smear cheese samples of various types, 15.8% contained organisms of the genus Listeria, 6.4% of the samples were contaminated with L. monocytogenes, 10.6% with L. innocua, and 1.2% with L. seeligeri. Six cheese samples contained two or more Listeria species, including at least one L. monocytogenes isolate. The incidences of L. monocytogenes in cheeses from various countries were: Italy 17.4%, Germany 9.2%, Austria 10%, and France 3.3%. Listeria were found most frequently in soft and semi-soft cheese. Eight samples contained more than 100 L. monocytogenes cfu/cm2 cheese surface, 2 samples had counts above 10(4) cfu/cm2 cheese surface. Surprisingly, a higher incidence of L. monocytogenes was observed in cheeses made from pasteurized milk (8.0%) than in cheeses manufactured from raw milk (4.8%). Phage-typing of isolated Listeria strains clearly confirmed that (i) contaminations within dairy plants were persistent over a period of several weeks to months and (ii) that cross-contamination within the dairy plant is and important factor. Comparison of our data with past surveys seems to indicate that contamination of red smear soft cheese with L. monocytogenes has not decreased sufficiently over the past 15 years. It is therefore strongly recommended that these products are monitored carefully by cheese-making companies.     

Efficacy of direct plating media for recovering Listeria monocytogenes from foods

Studies were done to evaluate 14 direct plating media for their suitability to recover Listeria monocytogenes strain Scott A from pasteurized whole milk, chocolate ice cream mix, Brie cheese and raw cabbage. Healthy cells were inoculated into foods to achieve viable populations of 10(2), 10(4), or 10(6) cells/ml(g). Bind's Acriflavine Agar, Trypaflavine Nalidixic Acid Serum Agar, Listeria Transport Enrichment Agar, Doyle and Schoeni Selective Enrichment Agar (DSSEA) and Modified DSSEA were not suitable for recovering L. monocytogenes from milk and Brie cheese and were therefore not evaluated as direct plating media for recovering the organism from ice cream mix and cabbage. McBride Listeria Agar (MLA), Gum Base Nalidixic Acid Tryptone Soya Agar (GBNTSA), Modified Despierres Agar (MDA) and Modified MLA (MMLA) performed best for recovering all inoculum populations from milk and ice cream mix. Enumeration of L. monocytogenes on several test media was complicated by the growth of large numbers of background microflora present in cabbage and Brie cheese, especially at the lowest test inoculum (10(2)/g). Generally, complete recovery of L. monocytogenes from Brie cheese and cabbage was attained on media when the inoculum population was greater than or equal to 10(4) cells/g. For Brie cheese, MLA, MDA, MMLA and Dominguez Rodriguez Isolation Agar were superior for recovering L. monocytogenes, while GBNTSA, DLEA, MDA and MMLA were best for recovering the organism from cabbage. Results of this experiment indicate that direct-plating procedures, without prior enrichment, can successfully be utilized for recovering L. monocytogenes from foods such as pasteurized milk and ice cream mix which contain low populations of background microflora. However, recovery of L. monocytogenes from foods such as raw cabbage and Brie cheese, which contain high populations of other microorganisms, was not satisfactory using the direct-plating procedures evaluated in this investigation.     

Origins and levels of post pasteurization contamination of milk in the dairy and their effects on keeping quality

Bacterial post pasteurization contamination with psychrotrophic Gram-negative rods (GNR) was measured in commercial milks before and after transfer to retail containers. The tanks of pasteurized milk feeding the filling units contaminated milk less often (39% of samples) but usually at a higher level than the filling units (92% of samples). The number of GNR present had a considerable influence on the shelf life of milk, and the range found in commercially pasteurized milk was reflected in a wide range of shelf lives.     

Peanut allergen (Ara h 1) detection in foods containing chocolate

Inadvertent exposure to peanut in foods poses health risks for peanut-allergic individuals that can be reduced by improving detection systems for allergen contaminants in food products and manufacturing processes. Detection of peanut in chocolate has been especially difficult. We report the optimization of conditions for measuring a major peanut allergen, Ara h 1, in chocolate with the use of a two-site monoclonal antibody sandwich enzyme-linked immunosorbent assay. Ara h 1 was extracted from peanut in the presence or absence of chocolate with phosphate buffer, salt, and three dried milks (goat, soy, or nonfat) (0 to 25% wt/vol) for 15 min at 60 degrees C or for 2.5 h at room temperature. The best conditions for Ara h 1 extraction in the presence of chocolate were 5% nonfat dry milk for 2.5 h at room temperature. Spiking experiments of chocolate with peanut confirmed improvement of the extraction: Ara h 1 was detected in extractions of 0.16 to 0.33% peanut in chocolate. Interestingly, the best conditions for Ara h 1 extraction were different for peanut alone than with chocolate, regarding time, temperature, and percentage of nonfat dry milk in the extraction buffer. In chocolate with peanut foods, the total Ara h 1 values were 10-fold higher than when products were extracted with phosphate buffer alone and could be up to 400-fold higher for individual foods. The dramatic improvement of Ara h 1 extraction should allow specific allergen monitoring in chocolate-containing food products and assessment of Ara h 1 exposure.     

Survival of Listeria monocytogenes in vanilla-flavored soy and dairy products stored at 8 degrees C

The survival of Listeria monocytogenes V37 in vanilla-flavored yogurt (low-fat and nonfat) and soy milk (low-fat and Plus) stored at 8 degrees C for 31 days was investigated. Commercial samples of yogurt and soy milk were used. These samples were inoculated with either 10(4) or 10(7) CFU of L. monocytogenes per ml. Sampling was carried out every 3 to 4 days initially and was then carried out weekly, for a total storage time of 31 days. Each time a sample was collected, the pH of the sample was measured. After 31 days, low-fat plain, low-fat vanilla, and nonfat plain yogurt samples inoculated with 10(4) CFU/ml showed 2.5-log reductions in viable cell populations, and nonfat vanilla yogurt showed a 3.5-log reduction. For yogurt inoculated with 10(7) CFU/ml, reductions of 2.5 log CFU/ml were observed for plain low-fat and nonfat yogurts, and reductions of 5 log CFU/ml were observed for vanilla-flavored low-fat and nonfat yogurts. In vanilla-flavored and plain low-fat and Plus soy milk samples, cell counts increased from 10(4) and 10(7) CFU/ml to 10(9) CFU/ml at 7 and 3 days of storage, respectively, at 8 degrees C. Coagulation in soy milk samples was observed when the cell population reached 10(9) CFU/ml. In soy milk, the L. monocytogenes population did not change for up to 31 days. Vanillin had an inhibitory effect on L. monocytogenes in yogurt but not in soy milk.     

Microbiological quality of retail cheeses made from raw, thermized or pasteurized milk in the UK

Two studies of retail fresh, ripened and semi-hard cheeses made from raw, thermized or pasteurized milk were undertaken in the UK during 2004 and 2005 to determine the microbiological quality of these products. Using microbiological criteria in European Commission Recommendations 2004/24/EC and 2005/175/EC, 2% of both raw, thermized (37/1819 samples) and pasteurized (51/2618 samples) milk cheeses were of unsatisfactory quality. Raw or thermized milk cheeses were of unsatisfactory quality due to levels of Staphylococcus aureus at 10(4)cfu g(-1), Escherichia coli at 10(5)cfu g(-1), and/or Listeria monocytogenes at 10(2)cfu g(-1), whereas pasteurized milk cheeses were of unsatisfactory quality due to S. aureus at 10(3)cfu g(-1) and/or E. coli at 10(3)cfu g(-1). Salmonella was not detected in any samples. Cheeses were of unsatisfactory quality more frequently when sampled from premises rated as having little or no confidence in management and control systems, and stored/displayed at above 8 degrees C. Raw or thermized milk cheeses were also more likely to be of unsatisfactory quality when they were unripened types, and pasteurized milk cheeses when they were: semi-hard types; from specialist cheese shops or delicatessens; cut to order. These results emphasize the need for applying and maintaining good hygiene practices throughout the food chain to prevent contamination and/or bacterial growth. Labelling of cheeses with clear information on whether the cheese was prepared from raw milk also requires improvement.     

Production of mortadella: behavior of Listeria monocytogenes during processing and storage conditions

The presence of Listeria monocytogenes in ready-to-eat meat products is cause of concern to the food industry as well as to health authorities. Studies were conducted to evaluate the presence of L. monocytogenes in mortadellas acquired at retail stores and to evaluate the fate of two levels of a L. monocytogenes pool spiked in two different formulations of the product, cooked under commercial conditions and stored at refrigeration (7°C) and room temperature (25°C). Among the samples collected at different retail stores, 26.7% harboured L. monocytogenes. Regarding to the fate of L. monocytogenes in mortadella, periodically, samples were taken and the surviving L. monocytogenes cells in the spiked products were counted by the MPN procedure. Populations of <0.35 MPN/g of L. monocytogenes were found in these samples. It could be concluded that the heat treatment was effective to reduce 3-log of L. monocytogenes independent of formulation or storage conditions.     

Survival, growth, and thermal resistance of Listeria monocytogenes in products containing peanut and chocolate

Outbreaks of listeriosis associated with the consumption of ready-to-eat foods have raised interest in determining growth, survival, and inactivation characteristics of Listeria monocytogenes in a wide range of products. A study was undertaken to determine the thermal tolerance of L. monocytogenes in a peanut-based beverage (3.1% fat), whole-fat (3.5%) milk, wholefat (4.0%) and reduced-fat (1.0%) chocolate milk, a chocolate-peanut spread (39% fat), and peanut butter (53% fat). The D60 degrees C value (decimal reduction time at 60 degrees C) in peanut beverage (3.2 min) was not significantly different (P > 0.05) than the D60 degrees C value in whole-fat milk (3.3 min) or whole-fat chocolate milk (4.5 min) but significantly lower (P < or = 0.05) than the D60 degrees C value in reduced-fat chocolate milk (5.9 min). The pathogen was significantly more resistant to heat when enmeshed in chocolate-peanut spread (water activity [aw] of 0.46; D60 degrees C = 37.5 min) and peanut butter (aw of 0.32; D60 degrees C = 26.0 min) than in liquid products. At 10 degrees C, the pathogen grew most rapidly in whole-fat chocolate milk and slowest in peanut beverage. At 22 degrees C, populations increased significantly within 12 and 16 h in whole-fat milk and reduced-fat chocolate milk, respectively, and within 8 h in whole-fat chocolate milk and peanut beverage. Initial populations (3.37 to 4.42 log CFU/g) of L. monocytogenes in chocolate-peanut spread and peanut butter adjusted to an aw of 0.33 and 0.65 declined, but the pathogen was not eliminated during a 24-week period at 20 degrees C. Survival was enhanced at reduced aw. Results indicate that a pasteurization process similar to that used for full-fat milk would be adequate to ensure the destruction of L. monocytogenes in peanut beverage. The pathogen survives for at least 24 weeks in chocolate-peanut spread and peanut butter at an aw range that encompasses that found in these products.     

Incidence and characterization of Listeria monocytogenes from domestic and imported foods in Korea

A total of 1,537 domestic and imported food products were examined for the incidence of Listeria monocytogenes between 1993 and 1997 in Korea. L. monocytogenes was detected using the U.S. Department of Agriculture isolation method. Isolated L. monocytogenes was confirmed by polymerase chain reaction with hly1 and hly2 primers designed from the listeriolysin O. Overall, 122 samples (7.9%) contained L. monocytogenes. The rate of isolation was 4.3% for beef, 19.1% for pork, 30.2% for chicken, 1.2% for shellfish, 4.4% for raw milk, 4.4% for frozen smoked mussels, and 6.1% for ice cream. No L. monocytogenes was found in pasteurized milk, pasteurized processed cheese, saltwater fish, dried seafoods, or ham. The overall incidence was lower than that reported in previous studies from other countries. Most isolates were serotype 1/2b except for chicken, in which serotype 1/2a was predominant. The serotyping results might imply the presence of food or geography-specific L. monocytogenes strains.     

Molecular Subtyping and Tracking of Listeria monocytogenes in Latin-Style Fresh-Cheese Processing Plants

Latin-style fresh cheeses, which have been linked to at least 2 human listeriosis outbreaks in the United States, are considered to be high-risk foods for Listeria monocytogenes contamination. We evaluated L. monocytogenes contamination patterns in 3 Latin-style fresh-cheese processing plants to gain a better understanding of L. monocytogenes contamination sources in the manufacture of these cheeses. Over a 6-mo period, 246 environmental samples were collected and analyzed for L. monocytogenes using both the Food and Drug Administration (FDA) method and the Biosynth L. monocytogenes detection system (LMDS). Finished cheese samples from the same plants (n = 111) were also analyzed by the FDA method, which was modified to include L. monocytogenes plating medium (LMPM) and the L. monocytogenes confirmatory plating medium (LMCM) used in the LMDS method. Listeria monocytogenes was detected in 6.3% of cheese and 11.0% of environmental samples. Crates, drains, and floor samples showed the highest contamination rates, with 55.6, 30.0, and 20.6% L. monocytogenes positive samples, respectively. Finished products and food contact surfaces were positive in only one plant. The FDA method showed a higher sensitivity than the LMDS method for detection of L. monocytogenes from environmental samples. The addition of LMPM and LMCM media did not further enhance the performance of the FDA method for L. monocytogenes detection from finished products. Molecular subtyping (PCR-based allelic analysis of the virulence genes actA and hly and automated ribotyping) was used to track contamination patterns. Ribotype DUP-1044A, which had previously been linked to a 1998 multistate human listeriosis outbreak in the United States, was the most commonly identified subtype (20/36 isolates) and was isolated from 2 plants. This ribotype was persistent and widespread in one factory, where it was also responsible for the contamination of finished products. We hypothesize that this ribotype may represent a clonal group with a specific ability to persist in food processing environments. While previous listeriosis outbreaks were linked to Latin-style fresh cheeses made from unpasteurized milk, the presence of this organism in pasteurized cheese products illustrates that persistent environmental contamination also represents an important source of finished product contamination.     

云南省牛乳中单核细胞增生李斯特氏菌的分布特征、耐药性及毒力研究

目的分析云南省牛乳中单核细胞增生李斯特氏菌(Listeria monocytogenes)的分布特征、耐药性及毒力现状。方法采集云南省主要奶产区牛乳样本,按照GB 4789.30—2016方法分离单核细胞增生李斯特氏菌疑似菌株,采用飞行时间质谱技术结合16SrDNA测序对疑似菌株进行鉴定;采用微量肉汤稀释法对分离菌株进行抗生素药敏实验;通过聚合酶链式反应(polymerasechainreaction,PCR)技术检测单核细胞增生李斯特氏菌中7种毒力基因的存在状况。结果158份样品共有4份检出单核细胞增生李斯特氏菌,检出率为2.53%;共分离获得8株菌。其中,生鲜乳中检出率为3.36%,巴氏杀菌乳中未检出。在生鲜乳中,荷斯坦牛乳、水牛乳、牦牛乳检出率分别为2.13%、0.00%和20.00%;标准化牛场、奶牛合作社和个体户的检出率分别为2.50%、1.51%和15.38%;手工挤奶和机械挤奶检出率为11.76%和1.96%。药敏实验结果显示8株菌均对青霉素耐药,对四环素、复方新诺明、红霉素耐药率分别为87.50%、75.00%和50.00%;多重耐药率为75.00%;但所有菌株对氨苄西林均敏感。8株菌对7种毒力基因的携带程度从12.50%~75.00%不等。结论云南省牛乳尤其生鲜乳中存在一定程度的单核细胞增生李斯特氏菌污染,且该菌具有普遍耐药性并携带一定程度的毒力基因,对消费者的安全具有潜在风险,应加强卫生监督管理。     

Incidence of Listeria monocytogenes in cheese manufacturing plants from the northeast region of São Paulo, Brazil

The incidence of Listeria monocytogenes in three cheese manufacturing plants from the northeastern region of S?o Paulo, Brazil, was evaluated from October 2008 to September 2009. L. monocytogenes was found in samples from two plants, at percentages of 13.3% (n = 128) and 9.6% (n = 114). Samples of raw and pasteurized milk, water, and Minas Frescal cheese were negative for L. monocytogenes, although the pathogen was isolated from the surface of Prato cheese and in brine from one of the plants evaluated. L. monocytogenes was also isolated from different sites of the facilities, mainly in non-food contact surfaces such as drains, floors, and platforms. Serotype 4b was the most predominant in the plants studied. The results of this study indicate the need for control strategies to prevent the dispersion of L. monocytogenes in the environment of cheese manufacturing plants.     

A survey of the microflora of raw and pasteurized milk and the sources of contamination in a milk processing plant in Addis Ababa, Ethiopia

The microorganisms present in raw and pasteurized milk and the sources of contamination in the milk after it had arrived at the processing plant in Addis Ababa were studied. The lowest count registered for raw milk samples was 4 X 10(7) cfu/ml while the highest was 1 X 10(9) cfu/ml. Pasteurized milk had mesophilic aerobic counts of 7 X 10(5) cfu/ml as it left the pasteurizing unit, but the population increased 2- to 4-fold as a result of subsequent contamination. Of the total counts in raw milk, psychrophilic, thermoduric and thermophilic organisms made up 98.1, 1.4 and 0.5% respectively. In pasteurized milk, the amounts were 53.0, 39.5 and 7.5% respectively. Samples of milk pasteurized in the laboratory contained only 74.5% thermoduric and 25.5% thermophilic organisms. The isolates mostly belonged to the genera Bacillus, Streptococcus, Lactobacillus, Arthrobacter, Alcaligenes, Aeromonas and Pseudomonas. Cocci were more predominant than rod-shaped bacteria. Of the rod-shaped bacteria, 73% were Gram-negative. The utensils holding the raw and pasteurized milk and the plastic sheets used for bagging the pasteurized milk contributed unusually high numbers of bacteria which were either thermoduric or thermophilic. More isolates were obtained from the pasteurized than the raw milk. The keeping quality of the pasteurized milk was found to be much lower than that of the laboratory-pasteurized milk.     

Prevalence and concentration of Listeria monocytogenes in sliced ready-to-eat meat products in the Hellenic retail market

The aim of this work was to estimate the prevalence and concentration of Listeria monocytogenes in packaged precut (slices or cubes) ready-to-eat (RTE) meat products available in the Hellenic retail market. Samples of these RTE meat products (n = 209) were taken from local supermarkets during a 3-month period and analyzed for the presence of L. monocytogenes with an automated enzymatic qualitative immunoassay followed by biochemical confirmation of positive results. The concentration of the pathogen in the positive samples was also determined. Seventeen samples (8.1%) were positive for L. monocytogenes. Eight (47.1%) of these 17 samples were from the same manufacturer; 36.4% of the products tested from this manufacturer were positive for L. monocytogenes. When bacon samples were not considered, the estimated prevalence of L. monocytogenes in sliced RTE meat products was much lower (3.1%). The L. monocytogenes populations in all positive samples were low, < or = 10 CFU/g. In 64.7% of the L. monocytogenes-positive samples, other Listeria species, including L. innocua and L. welshimeri, were also present at <10 to 690 CFU/g. These results indicate that L. monocytogenes is present in low numbers but is in a considerable proportion of the packaged precut RTE meat products that are sold in the Hellenic retail market. Cooked ham and bacon cut in cubes were the sample types most often contaminated with L. monocytogenes. The higher level of handling (e.g., cutting) associated with these products may further increase the risk of contamination with L. monocytogenes.     

PRESENCE OF LISTERIA IN MEXICAN CHEESES

The presence of Listeria was investigated in ripened cheeses (Chihuahua, Manchego) and fresh cheese (Panela) from street vendors and retail stores in Mexico City. Cheeses were tested for Listeria, pH, NaCl, moisture and fat. Listeria selective cold enrichment was used to recover Listeria from positive samples. Fresh cheese had the lowest pH and NaCl contents and the highest moisture Chihuahua and Manchego cheeses made with pasteurized milk were negative for Listeria. Panela cheese samples were the most contaminated. The presence of Listeria was 65% in fresh cheeses: Listeria murrayi 20% , Listeria inoccua 15% , Listeria grayi 15%, and Listeria monocytogenes 15%. L. monocytogenes serotypes 1/2a, 1/2b and 4b were isolated from positive samples.     

Competition of thermally injured listeria monocytogenes with a mesophilic lactic acid starter culture in milk for various heat treatments

Overnight tryptose broth cultures of three L monocytogenes strains were combined, centrifuged, suspended in 200 ml of tryptose phosphate broth, and heated at 56 degrees C for 20 min and at 64 degrees C for 2 min to obtain low-heat-injured (LHI) and high-heat-injured (HHI) cells, respectively, showing >99.6% injury. Flasks containing 200 ml of raw, low-heat-treated (56 degrees C for 20 min), high-heat-treated (64 degrees C for 2 min), pasteurized, and ultrahigh-temperature (UHT) milk were tempered to 31.1 degrees C and inoculated to contain 10(4) to 10(6) CFU/ml of LHI, HHI, or healthy L. monocytogenes cells and a commercial Lactococcus lactis subsp. lactis-Lactococcus lactis subsp. cremoris starter culture at levels of 0.5, 1.0, and 2.0%. Numbers of healthy and injured L. monocytogenes cells and starter organisms were determined using tryptose phosphate agar with or without 4.0% NaCl at selected intervals during 24 h of incubation at 31.1 degrees C. The presence of L. monocytogenes did not adversely affect the growth of the starter culture at any inoculation level. Overall, L. monocytogenes survived the 24-h fermentation period and grew to some extent. In starter-free controls. 76 to 81% of LHI cells and 59 to 69% of HHI cells were repaired after 8 h of incubation, with the lowest repair rates being observed for raw rather than heat-treated or pasteurized milk. Increased injury was observed for healthy L. monocytogenes cells at the 1.0 and 2.0% starter levels, with less injury seen for LHI and HHI cells. Raw and subpasteurized milk allowed less of a decrease in the percentage of injury and also showed higher numbers of injured cells than did pasteurized and UHT milks. These findings may have important implications for the survival of Listeria spp. in certain cheeses that can be prepared from raw or heat-treated milk.