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1.
Camembert-type cheese was produced from: raw bovine milk; raw milk inoculated with 2 or 4 log CFU/ml Listeria monocytogenes; raw milk inoculated with L. monocytogenes and subsequently pressure-treated at 500 MPa for 10 min at 20 °C; or uninoculated raw milk pressure-treated under these conditions. Cheeses produced from both pressure-treated milk and untreated milk had the typical composition, appearance and aroma of Camembert. Curd and cheese made from inoculated, untreated milk contained large numbers of L. monocytogenes throughout production. An initial inoculum of 1.95 log CFU/ml in milk increased to 4.52 log CFU/g in the curd and remained at a high level during ripening, with 3.85 log CFU/g in the final cheese. Pressure treatment inactivated L. monocytogenes in the raw milk at both inoculum levels and the pathogen was not detected in any of the final cheeses produced from pressure-treated milk. Therefore high pressure may be useful to inactivate L. monocytogenes in raw milk that is to be used for the production of soft, mould-ripened cheese.

Industrial relevance

This paper demonstrates the potential of high pressure (HP) for treatment of raw milk to be used in the manufacture of soft cheeses. HP treatment significantly reduced the level of Listeria monocytogenes in the raw milk and so allowed the production of safer non-thermally processed camembert-like soft cheese.  相似文献   

2.
A multiplex PCR procedure based on genes iap (coding for the invasion‐associated 60 kDa protein or p60) and hly (coding for listeriolysin O) of Listeria monocytogenes was to used to investigate the status of its contamination along the major milk‐processing environments. Duplex PCR amplified fragments of the iap gene at about 1.45 kb from all strains of major Listeria spp. tested and a 420 bp fragment of hly from L. monocytogenes reference strains. With triplex PCR, all L. monocytogenes strains exhibited a 420 bp fragment of hly as well as a 700 bp fragment of iap instead of its 1.45kb PCR product. The tentative L. monocytogenes isolates (n = 27) out of 566 samples of milk and milk‐processing environments in the PALCAM agar from cultures of buffered listeria enrichment broth were further subjected to both API and triplex PCR identification. Both methods identified the same 20 isolates as L. monocytogenes. The triplex PCR procedure detected as low as 3.2 × 101 cfu mL?1 of listerial cells even in the presence of L. innocua (108 cfu mL?1). It could detect 1.4 × 102 listerial cells mL?1 directly from milk artificially contaminated with the bacteria. Lower levels of L. monocytogenes cells in milk (1.45 × 101 to 1.45 × 100 cfu mL?1) could be detectable if the inoculated milk samples were cultured for 3–6 h. The bacterium was found not only in raw milk, sewage water and vessel surfaces but also in pasteurized milk. Copyright © 2005 Society of Chemical Industry  相似文献   

3.
Listeria monocytogenes is the etiologic agent of listeriosis responsible for severe and fatal infections in humans. Listeria contamination occurs quite often in a wide range of foods due to its ubiquitous nature. Isolates need to be characterized to a strain level for accurate diagnosis of Listeria infection, epidemiological studies, investigation of outbreaks and effective prevention and control of food-borne listeriosis. The purpose of this research was to evaluate the multiple-locus variable number of tandem repeat analysis (MLVA) for sub-typing L. monocytogenes isolates in pure cultures and in food matrices. Two multiplex PCR assays were formulated to amplify six specific loci using fluorescently-labeled primers; and the amplicons were analyzed by capillary electrophoresis. The MLVA method resulted in 34 unique DNA fingerprint patterns from 46 L. monocytogenes isolates of 10 serotypes which had 29 or 30 PFGE patterns with a single restriction enzyme and 34 AFLP patterns. The MLVA patterns of the 46 isolates remained unchanged in the presence of pre-enriched food matrices including sausage, ham, chicken, milk and lettuce. The MLVA method successfully typed L. monocytogenes strains spiked in cheese, roast beef, egg salad and vegetable samples after 48 h enrichment at the initial inoculation levels of 1-5 CFU per 25 g of food or higher. The limits of detection (typing) of the MLVA method were 103-104 CFU/mL of pre-enriched food broth when evaluated using post-spiked sausage, ham, chicken, milk and lettuce samples. The MLVA method was simple, highly discriminatory, and easy to perform with portable (numerical) results. To our knowledge, this is the first report that describes the application of the MLVA method directly to food samples and demonstrates the possibility to obtain rapid and accurate subtyping results before an isolate is obtained.  相似文献   

4.
A case of listeriosis was associated with the consumption of a soft cheese produced in England. Goats cheese and other products from the same food manufacturer were examined for the presence of Listeria over the following 11 months. Listeria monocytogenes was isolated from 16 of 25 cheese samples on retail sale, 12 of 24 cheese samples obtained directly from the factory, and from shelving within the plant. Phage-typing of 68 isolates of L. monocytogenes from cheese samples and the factory showed that 66 (97%) were indistinguishable from the strain isolated from the patient's cerebrospinal fluid and stool. L. monocytogenes was not isolated from seven goats milk or two yoghurt samples. Listeria innocua was isolated from 10 cheese samples, two of which contained no other species of Listeria. Levels of L. monocytogenes shortly after production were low (<10/g), but were higher (105–107 cfu/g) in six of the 16 cheese samples obtained from retail outlets. Multiplication of L. monocytogenes was demonstrated in cheeses contaminated at the factory and held at 4°C in the laboratory.  相似文献   

5.
A rapid and sensitive loop-mediated isothermal amplification (LAMP) assay was developed for detecting Listeria monocytogenes prfA in milk. The inclusivity of 23 L. monocytogenes and the exclusivity of 16 non-L. monocytogenes strains were both 100% in the assay. The limit of detection (LoD) of the LAMP assay in Listeria enrichment broth (LEB) was 2.22 CFU/mL after 12 h and 24 h of incubation. The LoDs of the LAMP assay in LEB with artificially contaminated milk (LEB-M) incubated for 12 h (2.22×101 CFU/mL) and 24 h (2.22 CFU/mL) were lower than those of the PCR and real-time PCR assays. Comparison of the LoDs in LEB with those in LEB-M showed that the LAMP assay was less influenced by the milk compounds than the real-time PCR assay. Our results indicate that the LAMP assay can be utilized as a potential screening tool for L. monocytogenes in milk.  相似文献   

6.
Bactivory by protozoa is a major factor that limits the number of bacteria in nature and may control the presence of Listeria monocytogenes. The effectiveness of Tetrahymena pyriformis destruction of L. monocytogenes was measured. Within 1 hr, 35–40 T. pyriformis cells ingested an average of 1,219 CFU of L. monocytogenes. Gentamicin was then added to kill un-ingested Listeria. In 24 hr, the recoverable bacteria were reduced at an exponential rate to undetectable levels (<1 per culture). A genetically diverse set of L. monocytogenes cultures all reduced Listeria recovery by the same degree. In assays without addition of gentamicin, numbers of attached L. monocytogenes cells were lessened from an average of log 6.5 CFU/2 ml culture to log 4.7 CFU/2 ml culture. T. pyriformis was capable of lowering numbers of both free-swimming and attached L. monocytogenes. This technology may have applications to control L. monocytogenes in food processing environments.  相似文献   

7.
Listeria monocytogenes Scott A and CA, were subjected at 23°C to hydrostatic pressures ranging from 2,380 to 3,400 atm and Vibrio parahaemolyticus T-3765-1 from 680 to 1,700 atm. For L. monocytogenes Scott A, pressurization in ultra-high temperature-processed (UHT) milk and raw milk appeared to provide a protective effect and lessened cell death as compared to pressurization in phosphate-buffered saline (100 mM, pH 7.0). A population of about 106 CFU/mL L. monocytogenes was killed by exposure to 3,400 atm within 80 min at 23°C in UHT milk. A population of about 106 CFU/mL V. parahaemolyticus was killed by exposure to 1,700 atm within 10 min at 23°C in clam juice.  相似文献   

8.
The bacterial pathogen Listeria monocytogenes is responsible for listeriosis, a food-borne disease, which may result in severe illness and possible death. Large outbreaks of listeriosis have been associated with food products including soft cheeses and ready to eat food products. Polymerase chain reaction (PCR) is a molecular identification method for food-borne pathogens; however, a drawback of this method is that false-positive or false-negative results may occur. To validate the accuracy of the PCR as a powerful molecular tool for pathogen detection, it is important that false-negative results be distinguishable from true-negative PCR results. The aim of this study was to design and include an internal amplification control (IAC) within the PCR to coamplify with L. monocytogenes in order to identify false-negative results of L. monocytogenes from ostrich meat and camembert cheese samples. The IAC had to be incorporated into the PCR without loss of specificity and sensitivity on the detection limit of L. monocytogenes and was developed and tested for use in a multiplex PCR detection system. A region of the pUC19 plasmid was selected as the IAC for this study. The optimal concentration at which pUC19 would coamplify with L. monocytogenes was determined to be 0.001 pg/μL. Following an enrichment procedure, the minimum number of organisms detected in a spiked food sample by the PCR was 8 CFU/mL L. monocytogenes; the same detection limit was attained when the pUC19 IAC was included in the PCR. An optimal pUC19 IAC concentration increased the reliability of the PCR for food diagnostic purposes.  相似文献   

9.
The aims of this work were (a) to select the minimum concentrations of two bacteriocin-producing lactobacilli strains to reduce Listeria monocytogenes growth in whole milk, (b) to evaluate the individual and combined application of these chosen lactobacilli concentrations as protective culture in this food and, (c) to estimate if lactobacilli growth caused undesirable changes in the visible characteristics of milk. The selected minimum lactobacilli concentration with antilisterial activity was 106 CFU ml−1 for both lactobacilli. Although a high initial concentration of L. monocytogenes (104 CFU ml−1) was added to milk, a notable reduction on listerial counts (2–4 log CFU ml−1) was achieved in comparison to the control. These lactobacilli showed a good adaptability in milk reaching optimal counts and not causing undesirable visible changes. In conclusion, the use of these lactobacilli could be considered as a bio-strategy potentially effective to limit the contamination by L. monocytogenes in certain milk-derived products.  相似文献   

10.
A multiplex PCR assay was developed by targeting ‘16S rRNA’ and ‘hly’ genes for detection of Listeria or Listeria monocytogenes in dairy foods on the basis of amplification of 1200 and 713 bp products, respectively. The assay conditions were optimized to make it truly rapid and to cut down the cost. The authenticity of the multiplex PCR was ascertained by using Nested PCR targeted against internal region of ‘hly’ gene that produced an amplified product of 188 bp. The multiplex PCR assay was found to be specific for detection of L. monocytogenes only since none of the non-listerial cultures gave positive signal. The sensitivity of the multiplex PCR was limited to 10 ng pure DNA and 1–10 cells of L. monocytogenes after 4–6 h enrichment in Listeria enrichment broth. When applied to 20 raw milk and 10 pasteurized milk samples, L. monocytogenes could not be detected in any of the samples by the multiplex PCR assay. This assay could find potential application in dairy industry for monitoring dairy foods for this high risk food pathogen on routine basis.  相似文献   

11.
Listeria monocytogenes was subjected to ultra high hydrostatic pressure (UHHP) treatments from 200 to 700 MPa at 25 °C in broth, raw milk, peach juice and orange juice. Survivor curves showed that cell death increased as pressure increased. After 10 min pressure treatment at 400 MPa reductions of about 2.09 and 2.76 log CFU mL?1 in aerobic bacteria and L. monocytogenes, respectively, were produced in raw milk, this increased to 5.09 and 6.47 log CFU mL?1, respectively, at 600 MPa. Death of bacteria at UHHP treatment was greater in orange juice than peach juice, and in peach juice than milk. Listeria monocytogenes was more sensitive to increased pressure than increased pressurization time. Injury of L. monocytogenes occurred from 0 to 100%. Factors effecting the rate of microbial inactivation are: pressure, age of cell, composition of medium, and pressurization time. UHHP inactivation can be used to extend shelf life and increase food quality during storage, and may also contribute to inactivation of L. monocytogenes.  相似文献   

12.
The objective of this study was to evaluate the inhibitory effect of grape seed extract (GSE), green tea extract (GTE), nisin and their combinations (nisin with either GSE or GTE) against Listeria monocytogenes. The inhibitory effect of these natural compounds was evaluated in phosphate buffer solution (PBS) medium containing approximately 109 colony‐forming units (CFU/mL) of L. monocytogenes. The effectiveness of these compounds in a meat model system was evaluated by surface inoculation (approximately 106 CFU/g) of L. monocytogenes onto turkey frankfurters. The inoculated frankfurters were dipped into soy protein film‐forming solutions with and without the addition of antimicrobial agents (GSE 1% or GTE 1% or nisin 10000 IU or combinations). Samples were stored at either 4 °C or 10 °C. The inhibitory effects of edible coatings were evaluated on a weekly basis for 28 d. The greatest inhibitory effect was observed in the PBS medium containing GSE (1%) and nisin (10000 IU/mL), which caused a 9‐log cycle reduction of L. monocytogenes population after 3 h incubation at 37 °C. In the meat system, the L. monocytogenes population (7.1 CFU/g) was decreased by more than 2 log cycle after 28 d at 4 °C and 10 °C, in the samples containing nisin (10000 IU) combined with either GSE (1%) or GTE (1%). This research has demonstrated that the use of an edible film coating containing both nisin and natural extracts is a promising means of controlling the growth and recontamination of L. monocytogenes on ready‐to‐eat meat products.  相似文献   

13.
ABSTRACT: This study was to develop an antimicrobial bottle coating method to reduce the risk of outbreaks of human listeriosis caused by contaminated liquid foods. Liquid egg white and skim milk were inoculated with Listeria monocytogenes Scott A and stored in glass jars that were coated with a mixture of polylactic acid (PLA) polymer and nisin. The efficacy of PLA per nisin coating in inactivating L. monocytogenes was investigated at 10 and 4 °C. The pathogen grew well in skim milk without PLA/nisin coating treatments, reaching 8 log CFU/mL after 10 d and then remained constant up to 42 d at 10 °C. The growth of Listeria at 4 °C was slower than that at 10 °C, taking 21 d to obtain 8 log CFU/mL. At both storage temperatures, the PLA coating with 250 mg nisin completely inactivated the cells of L. monocytogenes after 3 d and throughout the 42-d storage period. In liquid egg white, Listeria cells in control and PLA coating without nisin samples declined 1 log CFU/mL during the first 6 d at 10 °C and during 28 d at 4 °C, and then increased to 8 or 5.5 log CFU/mL. The treatment of PLA coating with 250 mg nisin rapidly reduced the cell numbers of Listeria in liquid egg white to undetectable levels after 1 d, then remained undetectable throughout the 48 d storage period at 10 °C and the 70 d storage period at 4 °C. These data suggested that the PLA/nisin coating treatments effectively inactivated the cells of L. monocytogenes in liquid egg white and skim milk samples at both 10 and 4 °C. This study demonstrated the commercial potential of applying the antimicrobial bottle coating method to milk, liquid eggs, and possibly other fluid products.  相似文献   

14.
Food safety is a critical factor in the production of farmhouse cheese. In Ireland the varieties of farmhouse cheese produced reflect a much broader range than those produced commercially and some of these cheese varieties are associated with greater microbiological risk. These include cheese produced from unpasteurised milk and soft ripened cheese such as mould or smear-ripened cheeses which have high pH and relatively short ripening times. The aim of this study was to determine the microbiological quality of farmhouse cheeses in Ireland. Three hundred and fifty one cheese samples, from 15 cheese producers, were analysed for microbiological quality on a monthly basis throughout the year. The analyses included enumeration of Escherichia coli, Staphylococcus aureus and Listeria monocytogenes (using the relevant agars) and enrichment for L. monocytogenes. The cheeses selected were produced from ovine, caprine and bovine milk. Both unpasteurised and pasteurised milk cheeses were sampled and these included hard, semi-hard and soft cheeses, internal/external mould-ripened and smear-ripened cheeses and the cheeses represented different geographic regions. Of the cheeses tested, 94% were free of L. monocytogenes, all were within the EU limits for E. coli and only one cheese variety had S. aureus levels above the recommended numbers for the first 6 months of the year. Due to a modified production process the numbers were within the guidelines for the second six months. The results indicate that Irish farmhouse cheeses are of a high microbiological quality.  相似文献   

15.
Isolation of Listeria spp. from 34 milk samples was compared by direct selective enrichment (FDA method) and two-stage enrichment techniques. Nineteen were Listeria-positive; 17 yielding one species only, either L. innocua or L. monocytogenes, and 2 both species. Direct enrichment and two-stage enrichment were equally effective at isolating Listeria spp. The number of isolations of L. monocytogenes was similar by both methods, but direct enrichment for more than 24 hr and use of the KOH step did not enhance the number of isolations.  相似文献   

16.
BACKGROUND: Commercially available fruits and raw and ready‐to‐eat vegetables (n = 445) were examined for aerobic, coliform, and yeast and mould counts using normalised methods. Listeria spp., Listeria monocytogenes and Salmonella spp. were detected by real time polymerase chain reaction (QPCR) after enrichment. RESULTS: Aerobic plate counts ranged from < 10 to > 109 colony‐forming units (CFU) g?1, with the lowest and highest counts recorded for fruits and sprouts respectively. The highest incidence level of coliforms was found in ready‐to‐eat vegetables, with up to 65.7% of samples containing from 5 to 9 log10CFU g?1. Yeasts and moulds showed their highest incidence level between 5 and 6 log10 CFU g?1, with an overall range from < 2 to 9 log10 CFU g?1. Salmonella spp., Listeria spp. and L. monocytogenes were detected in 0.67, 2.7 and 0.9% respectively of the total samples examined. CONCLUSION: The samples analysed can be gathered into two main groups, one showing low microbial counts (fruits) and a second group (raw whole leaves and roots and packed ready‐to‐eat vegetables) with higher microbial contamination. Although incidence levels of pathogenic bacteria reported here are in the lower range of those reported elsewhere, positive detections highlight the importance of good hygienic measures throughout the whole food chain. Copyright © 2007 Society of Chemical Industry  相似文献   

17.
The aim of this study was to verify the effectiveness of the commercially available anti-Listeria phage preparation LISTEXP100 in reducing Listeria monocytogenes on ready-to-eat (RTE) roast beef and cooked turkey in the presence or absence of the chemical antimicrobials potassium lactate (PL) and sodium diacetate (SD). Sliced RTE meat cores at 4 and 10 °C were inoculated with cold-adapted L. monocytogenes to result in a surface contamination level of 103 CFU/cm2. LISTEXTMP100 was applied at 107 PFU/cm2 and samples taken at regular time intervals during the RTE product's shelf life to enumerate viable L. monocytogenes. LISTEXP100 was effective during incubation at 4 °C with initial reductions of L. monocytogenes of 2.1 log10 CFU/cm2 and 1.7 log10 CFU/cm2, respectively, for cooked turkey and roast beef without chemical antimicrobials (there was no significant difference to the initial L. monocytogenes reductions in the presence of LISTEXTMP100 for cooked turkey containing PL and roast beef containing SD-PL). In the samples containing no chemical antimicrobials, the presence of phage resulted in lower L. monocytogenes numbers, relative to the untreated control, of about 2 log CFU/cm2 over a 28-day storage period at 4 °C. An initial L. monocytogenes cell reduction of 1.5 log10 CFU/cm2 and 1.7 log10 CFU/cm2, respectively, for cooked turkey and roast beef containing no chemical antimicrobials was achieved by the phage at 10 °C (abusive temperature). At this temperature, the L. monocytogenes cell numbers of samples treated with LISTEX™ P100 remained below those of the untreated control only during the first 14 days of the experiment for roast beef samples with and without antimicrobials. On day 28, the L. monocytogenes numbers on samples containing chemical antimicrobials and treated with LISTEXTMP100 stored at 4 and 10 °C were 4.5 log10 CFU/cm2 and 7.5 log10 CFU/cm2, respectively, for cooked turkey, and 1.2 log10 CFU/cm2 and 7.2 log10 CFU/cm2, respectively, for roast beef. In both cooked turkey samples with and without chemical antimicrobials stored at 10 °C, the phage-treated samples had significantly lower numbers of L. monocytogenes when compared to the untreated controls throughout the 28-day storage period (P < 0.0001). For roast beef and cooked turkey containing chemical antimicrobials treated with LISTEXTMP100 and stored at 4 °C, no more than a 2 log CFU/cm2 increase of L. monocytogenes was observed throughout the stated shelf life of the product. This study shows that LISTEXP100 causes an initial reduction of L. monocytogenes numbers and can serve as an additional hurdle to enhance the safety of RTE meats when used in combination with chemical antimicrobials.  相似文献   

18.
The aim of this study was to determine the growth kinetics of Listeria monocytogenes, with and without cold‐adaption, on fresh‐cut cantaloupe under different storage temperatures. Fresh‐cut samples, spot inoculated with a 4‐strain cocktail of L. monocytogenes (~3.2 log CFU/g), were exposed to constant storage temperatures held at 10, 15, 20, 25, or 30 °C. All growth curves of L. monocytogenes were fitted to the Baranyi, modified Gompertz, and Huang models. Regardless of conditions under which cells grew, the time needed to reach 5 log CFU/g decreased with the elevated storage temperature. Experimental results showed that there were no significant differences (P > 0.05) in the maximum growth rate k (log CFU/g h?1) and lag phase duration λ (h) between the cultures of L. monocytogenes with or without previous cold‐adaption treatments. No distinct difference was observed in the growth pattern among 3 primary models at various storage temperatures. The growth curves of secondary modeling were fitted on an Arrhenius‐type model for describing the relationship between k and temperature of the L. monocytogenes on fresh‐cut cantaloupe from 10 to 30 °C. The root mean square error values of secondary models for non‐ and cold‐adapted cells were 0.018, 0.021, and 0.024, and 0.039, 0.026, and 0.017 at the modified Gompertz, Baranyi, and Huang model, respectively, indicating that these 3 models presented the good statistical fit. This study may provide valuable information to predict the growth of L. monocytogenes on fresh‐cut cantaloupes at different storage conditions.  相似文献   

19.
ABSTRACT: Listeria monocytogenes and Salmonella typhimurium are major bacterial pathogens associated with poultry products. Ally isothiocyanate (AITC), a natural antimicrobial compound, is reportedly effective against these pathogenic organisms. A device was designed for the controlled release of AITC with modified atmosphere packaging (MAP), and then evaluated for its ability to control the growth of L. monocytogenes and S. typhimurium on raw chicken breast during refrigerated storage. In order to obtain controlled release during the test period, a glass vial was filled with AITC and triglyceride. It was then sealed using high-density polyethylene film. The release of AITC was controlled by the concentration (mole fraction) of AITC in the triglyceride and by the AITC vapor permeability through the film. The fresh chicken samples were inoculated with one or the other of the pathogens at 104 CFU/g, and the packages (with and without AITC-controlled release device) were flushed with ambient air or 30% CO2/70% N2 before sealing, and then stored at 4 °C for up to 21 d. The maximum reduction in MAP plus AITC (compared to MAP alone) was 0.77 log CFU/g for L. monocytogenes and 1.3 log CFU/g for S. typhimurium. The color of the chicken breast meat was affected by the concentration of AITC. Overall, a release rate of 0.6 μg/h of AITC was found to not affect the color, whereas at 1.2 μg/h of AITC the surface of the chicken was discolored.  相似文献   

20.
The study aimed at using essential oil (EO) alone or combined EO with nisin and low dose γ‐irradiation to evaluate their antibacterial effect against Listeria monocytogenes during storage of carrots at 4 °C. Minicarrots were inoculated with L. monocytogenes at a final concentration of approximately 7 log CFU/g. Inoculated samples were coated by nisin at final concentration of 103 International Unit (IU)/mL or individual mountain savory EO or carvacrol at final concentration of 0.35%, w/w) or nisin plus EO. The samples were then irradiated at 0, 0.5, and 1.0 kGy. The treated samples were kept at 4 °C and microbial analysis of samples were conducted at days 1, 3, 6, and 9. The results showed that coating carrots by carvacrol plus nisin or mountain savory plus nisin and then irradiating coated carrots at 1 kGy could reduce L. monocytogenes by more than 3 log at day 1 and reduced it to undetectable level from day 6. Thus, the combined treatments using nisin plus carvacrol or nisin plus mountain savory and irradiation at 1.0 kGy could be used as an effective method for controlling L. monocytogenes in minicarrots.  相似文献   

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