Behavior of Listeria monocytogenes in pasteurized milk during fermentation with lactic acid bacteria

The behavior of Listeria monocytogenes in pasteurized milk during fermentation with starter and nonstarter lactic acid bacteria was investigated. Pasteurized milk was co-inoculated with approximately 10(4) CFU/ml of L. monocytogenes and 10(6) CFU/ml of Lactococcus lactis, Lactococcus cremoris, Lactobacillus plantarum, Lactobacillus bulgaricus, or Streptococcus thermophilus. Inoculated milks were incubated at 30 degrees C or 37 degrees C for 24 to 72 h. Listeria monocytogenes survived and also grew to some extent during incubation in the presence of all starter cultures; however, inhibition ranged from 83 to 100% based on maximum cell populations. During incubation with L. bulgaricus and L. plantarum, L. monocytogenes was completely inactivated after 20 h and 64 h of incubation at 37 degrees C and 30 degrees C, respectively. The pH of the fermenting milks declined steadily throughout the fermentation periods and was approximately 4.2 at the conclusion of the experimental period regardless both of the starter culture and pathogen combination or the temperature of incubation.     

Effect of vanillin on the fate of Listeria monocytogenes and Escherichia coli O157:H7 in a model apple juice medium and in apple juice

The effects of vanillin on the fates of Listeria monocytogenes and Escherichia coli O157:H7 at pH values between 3.5 and 4.5 were verified in a model apple juice (MAJ) medium and in apple juice incubated at 4 or 15 degrees C. Viable E. coli O157:H7 cells were recovered from MAJ for up to 10 days, but L. monocytogenes did not survive at pH 3.5. Supplementation with 40 mm vanillin exerted a lethal effect that was species, concentration, pH and temperature dependant. E. coli O157:H7 was more sensitive to vanillin than L. monocytogenes, and viable cells could not be recovered after 2 days incubation at either temperature. L. monocytogenes and E. coli O157:H7 were inoculated (10(5) cfu/ml) in pH adjusted (pH 4.00) or unadjusted (pH 3.42) juice from Granny Smith apples that was supplemented with 40 mm vanillin. Neither species were recovered after 3 days incubation at 4 or 15 degrees C. These findings indicate that vanillin could be useful as a preservative for minimally processed apple products.     

Inhibitory combinations of nisin, sodium chloride, and pH on Listeria monocytogenes ATCC 15313 in broth by an experimental design approach

The influence of pH (5.0-8.2), NaCl concentrations (0-6% w/v), and incubation time (0-24 h) on the inhibitory activity of nisin (0-100 I.U./ml) against Listeria monocytogenes (10(3) cfu/ml) was studied using the Doehlert experimental design and was confirmed by kinetic experiments. Predicted values were in agreement with experimental values. Experiments were carried out at 22 degrees C in reconstituted TSB-YE1 broth with or without NaCl. Nisin had an immediate pH-dependent bactericidal effect, which increased with decreasing pH values. In modified TSB-YE1 broth without NaCl, the bactericidal efficacy of nisin (50 I.U./ml) was maximum at pH 6.6, with no L. monocytogenes survivors until 120 h at 22 degrees C. Nisin (50 I.U./ml) action decreased in the presence of NaCl, with a minimal inhibitory effect between 2 and 4%. This partially protective effect was cancelled at higher levels of nisin.     

Antagonism between Listeria monocytogenes and lactococci during fermentation of products from ultrafiltered skim milk.

Tyndallized samples of unfiltered skim milk and retentate (concentrated fivefold or twofold by volume) and permeate from UF skim milk were inoculated with 5.5 x 10(3) to 1.5 x 10(5) cfu/ml of Listeria monocytogenes strains California or V7 together with 4 x 10(7) to 2.3 x 10(8) cfu/ml of mesophilic lactic acid bacteria. Numbers of L. monocytogenes (McBride Listeria agar) and lactic acid bacteria (all purpose Tween agar) were determined after 0, 6, 12, 24, 30, and 36 h of incubation at 30 degrees C. Lactic acid bacteria significantly inhibited or inactivated L. monocytogenes in all three products. Inactivation was greater in permeate (6.77 orders of magnitude) than in unfiltered skim milk (3.67 orders of magnitude) or in retentate (4.21 orders of magnitude). Degree of inactivation in retentate was related to the extent of concentration. Inactivation was not complete, and L. monocytogenes survived in these products during fermentation for up to 36 h. When fermented products were refrigerated (4 degrees C), L. monocytogenes survived for 4 to 6 wk in skim milk, 3 to 5 wk in retentate, and 1 wk in permeate. At refrigeration temperature, length of survival was dependent on type of product and strain of the pathogen.     

Bactericidal effect of enterocin 4 on Listeria monocytogenes in a model dairy system.

Enterococcus faecalis INIA 4 produced the bacteriocin enterocin 4 during growth in raw ewe's milk at 30 degrees C. Enterocin activity reached 2,200 to 3,600 AU/ml after 8 h, with a 1 to 8% (vol/vol) level of inoculum from an 18-h culture. An enterocin activity of 500 AU/ml significantly decreased counts of Listeria monocytogenes Ohio when incubated for 6 h in a model system consisting of filtrates from cultures of E. faecalis INIA 4 in raw ewe's milk, at pH 6.0 and 30 degrees C. However, an enterocin activity of 2,400 AU/ml was needed in the same conditions to significantly decrease counts of L. monocytogenes Scott A. All 22 wild L. monocytogenes strains isolated from ewe's milk and tested were inhibited by a filtrate containing 400 AU/ml of enterocin 4. Incubation in the filtrate for 6 h significantly lowered counts of 16 L. monocytogenes strains, and incubation for 24 h, counts of 21 strains.     

Comparative survival of Salmonella typhimurium DT 104, Listeria monocytogenes, and Escherichia coli O157:H7 in preservative-free apple cider and simulated gastric fluid

This study compared the survival of three-strain mixtures (ca. 10(7) CFU ml(-1) each) of Salmonella typhimurium DT104, Listeria monocytogenes, and Escherichia coli O157:H7 in pasteurized and unpasteurized preservative-free apple cider (pH 3.3-3.5) during storage at 4 and 10 degrees C for up to 21 days. S. typhimurium DT104 populations decreased by <4.5 log10 CFU ml(-1) during 14 days storage at 4 and 10 degrees C in pasteurized cider, and by > or =5.5 log10 CFU ml(-1) during 14 days in unpasteurized cider stored at these temperatures. However, after 7 days at 4 degrees C, the S. typhimurium DT104 populations had decreased by only about 2.5 log10 CFU ml(-1) in both pasteurized and unpasteurized cider. Listeria monocytogenes populations decreased below the plating detection limit (10 CFU ml(-1)) within 2 days under all conditions tested. Survival of E. coli O157:H7 was similar to that of S. typhimurium DT104 in pasteurized cider at both 4 and 10 degrees C over the 21-days storage period, but E. coli O157:H7 survived better (ca. 5.0 log10 CFU ml(-1) decrease) than S. typhimurium DT104 (> 7.0 log10 CFU ml(-1) decrease) after 14 days at 4 degrees C in unpasteurized cider. In related experiments, when incubated in simulated gastric fluid (pH 1.5) at 37 degrees C, S. typhimurium DT104 and L. monocytogenes were eliminated (5.5-6.0 log10 CFU ml(-1) decrease) within 5 and 30 min, respectively, whereas E. coli O157:H7 concentrations decreased only 1.60-2.80 log10 CFU ml(-1) within 2 h.     

Growth-Death Kinetics of Listeria Monocytogenes in Strained Yogurt

In this research, viability of Listeria monocytogenes was studied in strained yogurt (Turkish name: Torba yogurt) samples prepared in laboratory conditions and contaminated with various levels of L. monocytogenes before and after incubation. When strained yogurt samples (pre-incubation inoculation, A1 and post-incubation inoculation, B1) were contaminated with a 10⁶ cfu/ml level of L. monocytogenes, their pH value reached to 4.08, and the presence of L. monocytogenes was defined only by using the enrichment method at the 30^th day of the storage at the pH of 3.55, since the resistance of the bacteria had been lost in those yogurts. However, L. monocytogenes was completely inhibited at the 7^th day of the storage for the strained yogurt samples (pre-incubation inoculation, A2 and post-incubation inoculation, B2) contaminated with the level of 10⁴ cfu/ml when the pH value was 3.58. As a result, it can be stated that viability of Listeria monocytogenes may not be possible if the pH of the yogurt is around the ≤3.59 after the 7^th day of the storage.     

Growth of Listeria monocytogenes in melon, watermelon and papaya pulps

Growth of Listeria monocytogenes in low-acid fruits (melon, watermelon and papaya) at different times of incubation and at temperatures of 10, 20 and 30 degrees C was studied. Fruit pulp portions with an average pH of 5.87, 5.50 and 4.87 for melon, watermelon and papaya, respectively, were obtained aseptically, homogenized, weighed and inoculated with suspensions (approximately 10(2) CFU/g) of L. monocytogenes. Generation times of 7.12, 13.03 and 15.05 h at 10 degrees C, 1.74, 2.17 and 6.42 h at 20 degrees C and 0.84, 1.00 and 1.16 h at 30 degrees C were obtained, respectively, for melon, watermelon and papaya. The results showed that L. monocytogenes grew in low-acid fruits at all tested temperatures, although growth was diminished, but not inhibited at 10 degrees C.     

The effects of temperature, pH, sodium chloride and sodium nitrite on the growth of Listeria monocytogenes.

An automated turbidimetric system using multiwelled plates was used to examine the effects of different combinations of NaCl (0.5-8.0% w/v), NaNO2 (0-400 micrograms/ml) pH (4.6-7.4) and temperature (5-30 degrees C) on the growth of Listeria monocytogenes in tryptone soya broth. The data presented clearly illustrate the combinations that permit visible growth of the organism. The ability of L. monocytogenes to grow at low pH levels was strongly influenced by incubation temperature as well as NaNO2 concentration. At 20 degrees C and below, no visible growth was detected, even with 50 micrograms/ml NaNO2 at pH 5.3 (or below) within 21 days. At pH 6.0 and above, NaNO2 had little effect in delaying visible growth except at higher concentrations and also at lower incubation temperatures.     

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.     

Growth of Listeria monocytogenes in egg salad and pasta salad formulated with mayonnaise of various pH and stored at refrigerated and abuse temperatures

This study investigated and modeled the behavior of Listeria monocytogenes in egg salad and pasta salad as affected by mayonnaise pH (3.8, 4.2, 4.6, and 5.0) and storage temperature (4, 8, and 12 degrees C). At each storage temperature, L. monocytogenes was able to grow in both salads regardless of the mayonnaise pH. The lag-phase durations (LPD) of L. monocytogenes in egg salad ranged from 33 to 85, 15 to 50, and 0 to 19 h, and the growth rates (GR) ranged from 0.0187 to 0.0318, 0.0387 to 0.0512, and 0.0694 to 0.1003 log(10)cfu/h at 4, 8, and 12 degrees C, respectively. The LPD of L. monocytogenes in pasta salad ranged from 210 to 430, 49 to 131, and 21 to 103 h, and GR ranged from 0.0118 to 0.0350, 0.0153 to 0.0418, and 0.0453 to 0.0718 log(10)cfu/h at 4, 8, and 12 degrees C, respectively. The growth of L. monocytogenes was more rapid in egg salad than in pasta salad, indicating that a better growth environment for L. monocytogenes existed in egg salad. In both salads, the LPD decreased and the GR increased as the storage temperature increased. Mathematical models and response surface plots describing the LPD and GR of L. monocytogenes in both salads as affected by the mayonnaise pH and storage temperature were developed. The models confirmed that the growth of L. monocytogenes in egg salad and pasta salad was primarily promoted by higher storage temperatures and, secondarily, by higher mayonnaise pH. The conditions under which the models may be applied to estimate the growth of L. monocytogenes in both salads were identified.     

Survival of Listeria monocytogenes during the manufacture and ripening of Swiss cheese.

Rindless Swiss cheese was made from a mixture of pasteurized whole and skim milk that was inoculated to contain 10(4) to 10(5) cfu of Listeria monocytogenes (strain Ohio, California, or V7)/ml. During clotting of milk, numbers of L. monocytogenes remained nearly unchanged. When the curd was heated gradually to attain the cooking temperature (50 degrees C), numbers of L. monocytogenes increased by approximately 40 to 45% over those in inoculated milk. Cooking curd at 50 degrees C (122 degrees F) for 30 to 40 min resulted in resilient curd having a pH of 6.40 to 6.45 and decreased L. monocytogenes by 48% compared with numbers of the pathogen in inoculated milk. After curd was pressed under whey, numbers of L. monocytogenes increased by approximately 52% over those in inoculated milk and reached their maxima at the end of this stage. A sharp decrease in numbers of L. monocytogenes occurred during brining of cheese blocks (7 degrees C for 30 h). The population of L. monocytogenes continued to decrease during cheese ripening. Average D values for strains California, Ohio, and V7 were 29.2, 24, and 22.5 d, respectively. Listeria was not detected (direct plating, and cold enrichment) after 80, 77, and 66 d of ripening of Swiss cheese made from milk inoculated with strains California, Ohio, and V7, respectively. Thus, Swiss cheese made in this study did not permit extended survival of L. monocytogenes.     

Antibacterial effect of water-soluble tea extracts on foodborne pathogens in laboratory medium and in a food model

The microbial inhibition of foodborne pathogens was determined in brain heart infusion broth with 10% (wt/vol) water-soluble extracts of green, jasmine, black, dungglre, and oolong tea against Escherichia coli O157:H7, Salmonella enterica serovar Enteritidis, Listeria monocytogenes, and Staphylococcus aureus. The mixed culture (approximately 6.0 log CFU/ml), which was composed of the four pathogens, was inoculated into brain heart infusion broth with and without tea extracts. After incubation at 35 degrees C for 0, 1, 3, and 5 days, proper dilution of each sample was spiral plated on each selective agar. Viable cell counts were performed after incubation at 35 degrees C for 24 to 36 h. Green, jasmine, and black tea exhibited an approximately 5.0 log suppression of S. aureus compared with the control from days 1 to 5. Green and jasmine tea also suppressed the growth of L. monocytogenes by approximately 3.0 log CFU/ml on day 5. In contrast, no tea extracts inactivated E. coli O157:H7 and Salmonella Enteritidis. Based on the result in liquid medium, green and jasmine teas of 0.1% (vol/wt) were individually evaluated for their antimicrobial activity against L. monocytogenes and S. aureus in a food model (ground beef) stored at 7 degrees C for 0, 1, 3, 5, and 7 days. Viable cell counts of total bacteria, L. monocytogenes, and S. aureus in ground beef were not significantly different among green and jasmine tea and the control.     

Biosynthesis of exopolysaccharide by a Bacillus licheniformis strain isolated from ropy cider

Listeria monocytogenes accumulates low molecular weight compounds (osmolytes, or compatible solutes) in response to chill stress. This response has been shown to be responsible, in part, for the chill tolerance of the species. Among the osmolytes tested to date, glycine betaine, gamma-butyrobetaine and carnitine display the strongest cryoprotective effect. These osmolytes are not synthesized in the cell and must be transported from the medium. In this study, the compatible solute accumulation profile of the food-borne pathogen L. monocytogenes was determined in balanced growth and stationary phase cultures grown in milk whey at 7 and 30 degrees C. In balanced growth cultures at 7 degrees C, glycine betaine (720 nmol/10(10) cfu) and carnitine (130 nmol/10(10) cfu) were the major osmolytes accumulated by wild-type L. monocytogenes 10403S, whereas carnitine (490 nmol/10(10) cfu) was the dominant osmolyte and glycine betaine was present in smaller amounts (270 nmol/10(10) cfu) in a mutant (L. monocytogenes LTG59) blocked in the major glycine betaine uptake system, glycine betaine porter II. In strain 10403S, glycine betaine and carnitine were present in eightfold and twofold excess at 7 degrees C compared to 30 degrees C; the respective ratios for strain LTG59 were 6 and 8. The intracellular concentration of osmolytes in stationary phase cultures at 7 degrees C was markedly reduced compared to that during balanced growth. Furthermore, at 4 degrees C, small but highly significant differences in growth were observed between strains. Strain LTG59 grew with a lag phase that was significantly longer, a generation time that was significantly greater and reached a final cell yield that was significantly lower than that of strain 10403S. The elevated accumulation of carnitine in the absence of glycine betaine porter II was insufficient to confer the magnitude of the cryoprotective effect displayed by the wild type.     

Fate of Escherichia coli O157:H7 and Listeria monocytogenes in strawberry juice and acidified media at different pH values and temperatures

Survival and growth of Escherichia coli O157:H7 and Listeria monocytogenes in strawberry juice and acidified media at different pH levels (pH 3.4 to 6.8) and temperatures were studied. Sterile strawberry juice (pH 3.6) and acidified trypticase soy broth (TSB) media (pH 3.4 to 6.8) were inoculated with approximately 6.7 log CFU/ml E. coli O157:H7 or 7.3 log CFU/ ml L. monocytogenes, incubated for 3 days at 4 and 37 degrees C. Bacterial levels were determined after 2 h, 1 day, and 3 days using surface plating nonselectively on tryptic soy agar and selectively on sorbitol MacConkey agar for E. coli O157:H7 or modified Oxford agar for L. monocytogenes. A spectrophotometer (660 nm) was also used to study growth inhibition of L. monocytogenes in different TSB and strawberry juice media (pH 3.4 to 7.3). E. coli O157:H7 survived well at pH values of 3.4 to 6.8 at 4 degrees C, but the number of injured cells increased as pH decreased and incubation time increased. At 37 degrees C, E. coli O157:H7 was inactivated at pH of < or = 3.6 but could grow at pH 4.7. L. monocytogenes was quickly injured at pH of < or = 4.7 within 2 h of storage at 4 degrees C and then was slightly and gradually inactivated as storage time increased. L. monocytogenes survived well at pH 6.8 at 4 degrees C and grew well at 37 degrees C. Growth of L. monocytogenes at 37 degrees C was inhibited in TSB by 1% citric acid and 0.5% malic acids at pH 3.4 or by 50% strawberry juice at pH 4.7. Bacterial injury and inactivation appeared to be induced by the acids in strawberry juice. The acids, pH value, temperature, and time were important factors for bacterial survival, inactivation, and growth in the media tested.     

Development of a lacticin 3147-enriched whey powder with inhibitory activity against foodborne pathogens.

The broad-spectrum bacteriocin lacticin 3147, produced by Lactococcus lactis DPC3147, is inhibitory to a wide range of gram-positive food spoilage and pathogenic organisms. A 10% solution of demineralized whey powder was fermented with DPC3147 at a constant pH of 6.5. The fermentate was spray dried, and the resulting powder exhibited inhibitory activity. The ability of the lacticin 3147-enriched powder to inhibit Listeria monocytogenes Scott A and Staphylococcus aureus 10 was assessed in buffer at both acidic (pH 5) and neutral (pH 7) pH. In addition, the ability of the powder to inhibit L. monocytogenes Scott A in an infant milk formulation was assessed. Resuspension of approximately 10(8) midexponential phase L. monocytogenes Scott A cells in a 10% solution of the lacticin 3147-enriched powder resulted in a 1,000-fold reduction in viable cells at pH 5 and pH 7 after 3 h at 30 degrees C. In the case of S. aureus 10, resuspension of 2.5 x 10(7) midexponential phase cells in a 15% solution of the lacticin 3147-enriched powder at pH 5 resulted in only a 10-fold reduction in viable cell counts, compared with a 1,000-fold reduction at pH 7, following incubation for 3 h at 30 degrees C. The use of the lacticin 3147 powder in an infant milk formulation resulted in greater than a 99% kill of L. monocytogenes within 3 h at 30 degrees C. These results suggest that this bioactive lacticin 3147 food ingredient may find applications in many different foods, including those with pH close to neutrality.     

Comparison of media and sampling locations for isolation of Listeria monocytogenes in queso fresco cheese

Listeriosis associated with Hispanic-style soft cheese is an ongoing public health concern. Although rapid detection methods based on molecular and immunological technologies have been applied successfully for detecting Listeria monocytogenes in foods, obtaining isolates of the pathogen is a critical procedure for epidemiologic studies and regulatory analysis. Oxford agar, a medium recommended by the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) to isolate L. monocytogenes from cheese, is unable to differentiate L. monocytogenes from other Listeria species. Hence, two selective isolation media, L. monocytogenes blood agar (LMBA) and Rapid 'L. mono agar (RLMA), were compared with Oxford agar for isolating L. monocytogenes from cheese. Queso fresco cheese was inoculated at 10(0) or 10(1) CFU/g with a five-strain mixture of L. monocytogenes or with the five-strain L. monocytogenes mixture and Listeria innocua. Cheese samples were stored at 21, 12, and 4 degrees C and Listeria counts were determined at 3, 7, and 10 days; 7, 10, 14, 21 days; and 2, 4, 8, and 12 weeks postinoculation, respectively. Surface and interior cheese samples as well as liquid exudate produced during storage were assayed individually to determine differences in Listeria contamination at different sampling locations. L. monocytogenes was more easily differentiated from L. innocua on RLMA than LMBA and Oxford agar. Similar L. monocytogenes counts (ca. 10(4) CFU/g) were obtained on the last sampling day on the surface and interior of cheese samples (P > 0.05) for all storage temperatures and both initial inoculation levels, but smaller cell numbers were detected in the exudate produced during storage. In addition, simultaneous inoculation of L. innocua with L. monocytogenes did not affect the final L. monocytogenes counts in the cheese. The amount of exudate released from the cheese and decrease of pH correlated with storage temperature. More exudate was produced and a greater decrease of pH occurred at 21 degrees C than at 12 or 4 degrees C. Our results indicate that RLMA is a suitable medium for isolating L. monocytogenes from queso fresco cheese. Higher counts of L. monocytogenes were obtained from surface and interior samples of cheese than from the exudate of the cheese during storage. In addition, pH may be a useful indicator of improperly stored queso fresco cheese.     

Survival and growth of Salmonella baildon in shredded lettuce and diced tomatoes, and effectiveness of chlorinated water as a sanitizer

An outbreak of salmonellosis associated with diced tomatoes occurred in the United States in 1999. Experiments were done to determine the efficacy of chlorine in killing Salmonella baildon, the causative serotype, inoculated onto shredded lettuce and diced tomatoes, and to determine survival characteristics of the organism on these produce items stored at 4 degrees C for up to 12 days and on tomatoes stored at 21 or 30 degrees C for up to 72 h. Populations of S. baildon in lettuce and tomatoes (pH 4.51 +/- 0.02) inoculated with 3.60 log10 and 3.86 log10 cfu/g, respectively, were reduced by less than 1 log when the produce was immersed for 40 s in a 120 or 200 microg/ml free chlorine solution. Produce inoculated with 0.60-0.86 log10 cfu/g was positive for the pathogen after treatment with 200 microg/ml chlorine. Initial populations of 3.28 and 3.40 log10 cfu/g of lettuce and tomatoes, respectively, decreased by about 2 log10 cfu/g during storage for 12 days at 4 degrees C. One of six samples of lettuce initially containing 0.28 log10 cfu of S. baildon per gram was positive after storage for 12 days, but the pathogen was not detected in tomatoes analyzed within 15 min of inoculation with 0.40 log10 cfu/g. While the number of viable cells decreased during storage at 4 degrees C, initial populations of 0.28 log10 cfu/g of shredded lettuce and 3.40 log10 cfu/g of diced tomatoes are not reduced to undetectable levels during storage at 4 degrees C for 12 days. Tolerance of S. baildon to an acidic pH (4.5) was not influenced by the pH (4.5, 5.8, or 7.2) of the medium in which it was grown, suggesting that this strain possesses unusual resistance to acid pH. The pathogen grew in diced tomatoes (pH 4.40 +/- 0.01) from an initial population of 0.79 log10 cfu/g to 5.32 and 7.00 log10 cfu/g within 24 h at 21 and 30 degrees C, respectively.     

Inactivation of Listeria monocytogenes Scott A 49594 in apple juice supplemented with cinnamon

Normal (pH 3.7) and adjusted (pH 5.0) pasteurized apple juice containing cinnamon (0, 0.1, 0.2, and 0.3%) was inoculated with Listeria monocytogenes Scott A 49594 at 10(4) CFU/ml and stored at 5 and 20 degrees C for 7 days. Counts on tryptic soy agar (TSA), modified Oxford (MOX) medium, and thin agar layer (TAL) were determined at 1 h and 1, 3, and 7 days. The TAL method (MOX medium overlaid with TSA) was used for the recovery of injured cells. In apple juice, both at normal and adjusted pH, with any doses of cinnamon, no L. monocytogenes (a 4.6-log CFU/ml reduction) was detected after 1 h of storage at both temperatures, and no growth occurred at any points of storage. Therefore, cinnamon by itself (regardless of pH) had a pronounced killing effect. A further enrichment step with brain heart infusion agar showed that L monocytogenes was completely inactivated in apple juice stored at 20 degrees C, except in pH 5.0 samples with 0.1% of cinnamon. The TAL method was as effective as TSA in recovering injured cells of L. monocytogenes. Cinnamon considerably inactivates L. monocytogenes in apple juice and thus enhances the safety of this product.     

Enumeration and growth of naturally occurring Listeria spp. in unpackaged ham

A total of 301 unpackaged retail ham samples were tested for the presence and number of Listeria spp. after 7 days at 5 degrees C to simulate domestic storage. Thirteen samples (4.3%) contained Listeria monocytogenes, with the highest count being 1.6 x 10(3)cfu g(-1). Thirteen samples contained other Listeria spp. Genotyping showed that only one L. monocytogenes isolate from the 14 tested was of a type previously identified in New Zealand human cases. Listeria-contaminated batches were incubated at 5 degrees C over approximately 3 weeks to assess the growth rate of natural contaminants. None contained L. monocytogenes, but growth occurred in one sample containing Listeria welshimeri and four containing Listeria innocua. Growth was usually slow at 0.002-0.004 log h(-1). In one sample, L. innocua grew at 0.02 log h(-1) although the maximum number reached was only 4.0-5.0 x 10(3)cfu g(-1). In five other samples little growth, if any, occurred. Growth of naturally occurring Listeria spp. at 5 degrees C was therefore generally slower than predicted by the Pathogen Modelling Programme (PMP) or did not occur.