Tag Archives: L. monocytogenes

Research – L. monocytogenes in a cheese processing facility: Learning from contamination scenarios over three years of sampling

Science Direct Eurofins Food Testing UK

The aim of this study was to analyze the changing patterns of Listeria monocytogenes contamination in a cheese processing facility manufacturing a wide range of ready-to-eat products. Characterization of L. monocytogenes isolates included genotyping by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Disinfectant-susceptibility tests and the assessment of L. monocytogenes survival in fresh cheese were also conducted. During the sampling period between 2010 and 2013, a total of 1284 environmental samples were investigated. Overall occurrence rates of Listeria spp. and L. monocytogenes were 21.9% and 19.5%, respectively. Identical L. monocytogenes genotypes were found in the food processing environment (FPE), raw materials and in products. Interventions after the sampling events changed contamination scenarios substantially. The high diversity of globally, widely distributed L. monocytogenes genotypes was reduced by identifying the major sources of contamination. Although susceptible to a broad range of disinfectants and cleaners, one dominant L. monocytogenes sequence type (ST) 5 could not be eradicated from drains and floors. Significantly, intense humidity and steam could be observed in all rooms and water residues were visible on floors due to increased cleaning strategies. This could explain the high L. monocytogenes contamination of the FPE (drains, shoes and floors) throughout the study (15.8%). The outcome of a challenge experiment in fresh cheese showed that L. monocytogenes could survive after 14 days of storage at insufficient cooling temperatures (8 and 16 °C). All efforts to reduce L. monocytogenes environmental contamination eventually led to a transition from dynamic to stable contamination scenarios. Consequently, implementation of systematic environmental monitoring via in-house systems should either aim for total avoidance of FPE colonization, or emphasize a first reduction of L. monocytogenes to sites where contamination of the processed product is unlikely. Drying of surfaces after cleaning is highly recommended to facilitate the L. monocytogenes eradication.

 

Research – Prevalence, Antibiotic Resistance and Genetic Diversity of Listeria monocytogenes Isolated From Retail Ready-To-Eat Foods in China

Science Direct

Listeria monocytogenes is a major foodborne pathogen that is well known as high mortality rate upon infected. This study aimed to investigate the prevalence of L. monocytogenes isolates from retail ready-to-eat (RTE) foods in China and characterize the isolates of L. monocytogenes by antibiotic resistance, serotyping, ERIC-PCR and REP-PCR subtyping analyses. From September 2012 to January 2014, a total of 364 retail RTE foods were obtained. Using the qualitative and quantitative methods, 25 samples (6.87%) were positive for L. monocytogenes. The identity of isolates of L. monocytogenes was confirmed by PCR. All 80 isolates in this survey were sensitive to penicillin and mezlocillin, the highest resistance is clindamycin (51.25%), followed by cephalothin (23.75%) and ampicillin (12.5%). Twenty-seven isolates were susceptible to all 14 tested antibiotics; seventeen isolates were resistant to more than two antibiotics, including six multiresistent strains resist to more than 10 antibiotics. L. monocytogenes isolates belonged to serovar types 1/2a (3a), 4b (4d, 4e), 1/2b (3b, 7) and 1/2c (3c). 29 L. monocytogenes isolates were selected by serotyping. At the relative similarity coefficient of 0.80, it grouped 29 isolates and 5 reference strains into 2 clusters and 3 singletons, 4 clusters and 1 singleton by ERIC-PCR and REP-PCR, respectively. Our study reflects the potential risk of L. monocytogenes infection in China. We also provide a comprehensive surveillance on its incidence on the RTE foods of L. monocytogenes and ensure more accurate treatment of human listeriosis with effective antibiotics.

New Zealand – Limits for Listeria in RTE Foods

FSANZ

Criteria for Listeria monocytogees in ready-to-eat foods

New criteria for Listeria monocytogenes in ready-to-eat (RTE) foods were gazetted in the Food Standards Code on 31 July 2014.
Two sets of criteria for L. monocytogenes now apply based on whether growth of L. monocytogenes will or will not occur in the RTE food:
  • RTE foods in which growth of L. monocytogenes will not occur (less than 100cfu/g).
  • RTE foods in which growth of L. monocytogenes will occur (not detected in 25g).
This approach is risk-based and has been informed by improved scientific knowledge about the factors affecting the risk of acquiring listeriosis.
It is consistent with international approaches, including Codex guidelines and recognises that L. monocytogenes is able to grow to high numbers in some foods and should not be present at detectable levels if the food is to be kept safe. In other foods, where we know the pathogen cannot grow, there is a possibility for occasional low level detections that will not affect the safety of the food.

Research – Listeria monocytogenes – Pet Foods – RTE Ham

Mary Ann Leibert

The Veterinary Laboratory Investigation and Response Network (Vet-LIRN), in collaboration with the Food Emergency Response Network (FERN) and its Microbiology Cooperative Agreement Program (MCAP) laboratories, conducted a study to evaluate the prevalence of selected microbial organisms in various types of pet foods. The goal of this blinded study was to help the Center for Veterinary Medicine prioritize potential future pet food–testing efforts. The study also increased the FERN laboratories’ screening capabilities for foodborne pathogens in animal feed matrices, since such pathogens may also be a significant health risk to consumers who come into contact with pet foods. Six U.S. Food and Drug Administration FERN MCAP laboratories analyzed approximately 1056 samples over 2 years. Laboratories tested for Salmonella, Listeria, Escherichia coli O157:H7 enterohemorrhagic E. coli, and Shiga toxin–producing strains of E. coli (STEC). Dry and semimoist dog and cat foods purchased from local stores were tested during Phase 1. Raw dog and cat foods, exotic animal feed, and jerky-type treats purchased through the Internet were tested in Phase 2. Of the 480 dry and semimoist samples, only 2 tested positive: 1 for Salmonella and 1 for Listeria greyii. However, of the 576 samples analyzed during Phase 2, 66 samples were positive for Listeria (32 of those were Listeria monocytogenes) and 15 samples positive for Salmonella. These pathogens were isolated from raw foods and jerky-type treats, not the exotic animal dry feeds. This study showed that raw pet foods may harbor food safety pathogens, such as Listeria monocytogenes and Salmonella. Consumers should handle these products carefully, being mindful of the potential risks to human and animal health.

Mary Ann Leibert

Ready-to-eat (RTE) meat and poultry products manufactured with natural or organic methods are at greater risk for Listeria monocytogenes growth, if contaminated, than their conventional counterparts due to the required absence of preservatives and antimicrobials. Thus, the objective of this study was to investigate the use of commercially available natural antimicrobials and postlethality interventions in the control of L. monocytogenes growth and recovery on a RTE ham product. Antimicrobials evaluated were cranberry powder (90MX), vinegar (DV), and vinegar/lemon juice concentrate (LV1X). Postlethality interventions studied were high hydrostatic pressure at 400 (HHP400) or 600 (HHP600) MPa, lauric arginate (LAE), octanoic acid (OA), and postpackaging thermal treatment (PPTT). Parameters evaluated through 98 days of storage at 4±1°C were residual nitrite concentrations, pH, aw, and viable L. monocytogenes on modified Oxford (MOX) media. On day 1, OA, 90MX, DV, and LV1X yielded lower residual nitrite concentrations than the control, whereas HHP400, HHP600, and LAE did not. LAE, HHP400, and OA reduced L. monocytogenes population compared to the control after 1 day of storage by 2.38, 2.21, and 1.73 log10 colony-forming units per gram, respectively. PPTT did not achieve a significant reduction in L. monocytogenes populations. L. monocytogenes recovered and grew in all postlethality intervention treatments except HHP600. 90MX did not inhibit the growth of L. monocytogenes, while DV and LV1X did. Results of this study demonstrate the bactericidal properties of HHP, OA, and LAE and the bacteriostatic potential of natural antimicrobial ingredients such as DV and LV1X against L. monocytogenes.

Research Articles – Listeria – Mustard Anti-Microbial – Anti-Microbial Resistance – Growth Model

Journal of Food Science

The antimicrobial activities of oriental mustard extract alone or combined with malic acid and EDTA were investigated against Salmonella spp. or Listeria monocytogenes at different temperatures. Five strain Salmonella or L. monocytogenes cocktails were separately inoculated in Brain Heart Infusion broth containing 0.5% (w/v) aqueous oriental mustard extract and incubated at 4 °C to 21 °C for 21 d. For inhibitor combination tests, Salmonella Typhimurium 02:8423 and L. monocytogenes 2–243 were individually inoculated in Mueller Hinton broth containing the mustard extract with either or both 0.2% (w/v) malic acid and 0.2% (w/v) EDTA and incubated at 10 °C or 21 °C for 10 to 14 d. Mustard extract inhibited growth of the L. monocytogenes cocktail at 4 °C up to 21 d (2.3 log10 CFU/mL inhibition) or at 10 °C for 7 d (2.4 log10 CFU/mL inhibition). Salmonella spp. viability was slightly, but significantly reduced by mustard extract at 4 °C by 21 d. Although hydrolysis of sinigrin in mustard extract by both pathogens was 2 to 6 times higher at 21 °C than at 4 °C to 10 °C, mustard was not inhibitory at 21 °C, perhaps because of the instability of its hydrolysis product (allyl isothiocyanate). At 21 °C, additive inhibitory effects of mustard extract with EDTA or malic acid led to undetectable levels of S. Typhimurium and L. monocytogenes by 7 d and 10 d, respectively. At 10 °C, S. Typhimurium was similarly susceptible, but combinations of antimicrobials were not more inhibitory to L. monocytogenes than the individual agents.

Science Direct

Listeria monocytogenes is an important foodborne pathogen. The aims of this study were to determine genetic relatedness of L. monocytogenes isolated from ready-to-eat (RTE) foods in Malaysia. L. monocytogenes isolates from RTE foods were characterized by multiplex-PCR serotyping, REP-PCR, BOX-PCR, RAPD, PFGE, virulotyping and antibiotyping. Of the 32 L. monocytogenes isolates analyzed, 21 (65.6%) were assigned to serogroup “1/2a, 3a”, seven (21.9%) serogroup “1/2c, 3c”, and four (12.5%) serogroup “4b, 4d, 4e”. All the L. monocytogenes harbored inlA, inlB, inlC and inlJ virulence genes. More than half (53%) L. monocytogenes isolates were resistant to penicillin G, followed by tetracycline (15.6%), amoxicillin-clavulanic acid (12.5%), vancomycin (9.4%) erythromycin (6.3%), clindamycin, streptomycin, kanamycin, and chloramphenicol (each 3.1%). REP-PCR, BOX-PCR, RAPD and PFGE generated 28 (D = 0.992), 31 (D = 0.998), 32 (D = 1), and 20 (D = 0.916) patterns, respectively. These results indicate that L. monocytogenes isolates from RTE food were heterogeneous. There was no correlation between antibiograms and serogroups or pulsotypes or PCR-typing and/or sources of isolates. Since different subtyping methods often give different discriminatory powers, the use of more than one subtyping approach is necessary in providing a more accurate picture of the genetic diversity of L. monocytogenes. In conclusion, L. monocytogenes isolates from RTE possess the internalin genes and are genetically diverse. Furthermore, the occurrence of resistant isolates belonging to epidemiologically important serogroups “1/2a, 3a” and “4b, 4d, 4e” in RTE foods is a matter of public health concern.

Science Direct

This study was performed to develop a predictive growth model of Listeria monocytogenes to ensure the safety of raw pork. The pork samples were inoculated with a cocktail of two L. monocytogenes strains ATCC 15313 and L13-2 isolated from pork and were stored at 5, 15, and 25 °C. Results were evaluated using the MicroFit program. To develop primary models, the Baranyi, modified Gompertz, and Logistic model equations were applied to the observed data. The mathematically predicted growth rate parameters were evaluated using the coefficient of determination (R2), bias factor (Bf), accuracy factor (Af), and mean square error (MSE). The Baranyi model, which showed an R2 of 0.998 and MSE of 0.006, was more suitable than the modified Gompertz and Logistic models. In validation study of secondary model, it appeared that MSE’s of specific growth rate (SGR) and lag time (LT) were relatively accurate and suitable for modeling the growth of L. monocytogenes. These values indicated that the developed models were acceptable for expressing the growth of microorganisms on raw pork, which can be applied to ensure the safety of meats and to establish standards for avoiding microbial contamination.

Research – Listeria Growth in Semi Soft Cheese

Science Direct

The objective of this study was to investigate the growth of Listeria monocytogenes in semi-soft rind washed cheese made from raw and pasteurised milk at different storage temperatures (4, 10 and 15 °C) over a 28 day period simulating storage following ripening. Changes in water activity (aw) and pH in cheeses were also monitored during storage. Response surface models were used to model the interaction of storage temperature and time on aw, pH and L. monocytogenes population. Growth curves were fitted using Baranyi, modified Gompertz and Logistic models at all storage temperatures for both cheeses, and model parameters were statistically analysed. In raw and pasteurised milk cheeses, all models showed a significant (P < 0.05) increase in the specific growth rate (SGR, Day−1) of L. monocytogenes with an increase in storage temperature. A higher SGR was observed for L. monocytogenes in pasteurised milk cheese (0.18–0.85 Day−1) compared to raw milk cheese (0.05–0.37 Day−1) at all storage temperatures studied. Response surface models indicated an increase in the L. monocytogenes population and pH with an increase in storage temperature. However, a decreasing trend in aw for both cheese types was observed. The predicted regression model parameters for both the raw and pasteurised milk cheese showed a high correlation coefficient R2 > 0.87. Overall, the L. monocytogenes population increased up to 3 log10 cfug−1 for both cheeses during storage following ripening. The fitted models confirmed different L. monocytogenes growth behaviour between raw and pasteurised milk cheeses, which could support the Food Business Operator in predicting growth during storage following ripening.

Research -Modeling the Growth of Listeria monocytogenes on Cut Cantaloupe, Honeydew and Watermelon

Science Direct

A recent outbreak linked to whole cantaloupes underscores the importance of understanding growth kinetics of Listeria monocytogenes in cut melons at different temperatures. Whole cantaloupe, watermelon, and honeydew purchased from a local supermarket were cut into 10 ± 1 g cubes. A four-strain cocktail of L. monocytogenes from food related outbreaks was used to inoculate fruit, resulting in ∼103 CFU/10 g. Samples were stored at 4, 10, 15, 20, or 25 °C and L. monocytogenes were enumerated at appropriate time intervals. The square root model was used to describe L. monocytogenes growth rate as a function of temperature. The model was compared to prior models for Salmonella and Escherichia coli O157:H7 growth on cut melon, as well as models for L. monocytogenes on cantaloupe and L. monocytogenes ComBase models. The current model predicts faster growth of L. monocytogenes vs. Salmonella and E. coli O157:H7 at temperatures below 20 °C, and agrees with estimates from ComBase Predictor, and a corrected published model for L. monocytogenes on cut cantaloupe. The model predicts ∼4 log CFU increase following 15 days at 5 °C, and ∼1 log CFU increase following 6 days at 4 °C. The model can also be used in subsequent quantitative microbial risk assessments.

Research – Brasil – Risk of Infection by Salmonella and Listeria from RTE Vegetables

Science DirectEurofins Food Testing UK

The current study was carried out to estimate the risks of infection due to consumption of RTE vegetables contaminated with Salmonella and Listeria monocytogenes in Brazil. The risk assessment model was composed of five different modules comprising the retail-consumption steps. Scenarios were simulated using prevalence and concentration levels reported in RTE vegetables in Brazil as well as considering values 10 times lower. In addition, scenarios in which temperature during transportation and storage are maintained below 5 °C were also evaluated. Models built in Excel spreadsheets were run (100,000 iterations) using @Risk software. The two outputs were risk of infection per month (probability of infection per month due to consumption of RTE vegetables) and number of infections per month (number of people that consumed RTE vegetables and get infected per month). The QMRA models predicted that the mean risk of Salmonella infection per month is 5.7E-03, while the mean risk of infection for L. monocytogenes was 8.1E-06 per month. The reduction of prevalence of Salmonella from 1.7% to 0.17% resulted in a decrease of risk of infection per month by about 6 times. In the case of L. monocytogenes, the reduction of prevalence from 2.2% to 0.22% resulted in decrease of risk of infection from 8.1E-06 to 1.0E-06. The risks and number of cases predicted in scenarios in which temperature was kept below 5 °C were reduced for both pathogens studied when compared to scenarios where this was not the case. The scenario where prevalence and concentration of pathogens was reduced and where temperature was <5 °C led to the lowest number of infections due by Salmonella and L. monocytogenes (187 and 3.3E-05 cases, respectively). The results suggest that effective mitigation strategies need to be adopted. The strict control of temperature during transportation, storage and consumption was more effective to reduce risk and number of cases due to L. monocytogenes than to Salmonella. More data is needed to improve the accuracy of risk assessment models developed.