Modelling the growth of Listeria monocytogenes in dynamic conditions

A recurrent neural network for the prediction of Listeria monocytogenes growth under pH and a(w) variable conditions was developed. The use of this model offered the possibility to take into account the consequences of the variations of the factors on L. monocytogenes growth. The effects of solutions, such as NaCl, acetic acid and NaOH, and their interactions on the response of L. monocytogenes cells were studied. Furthermore, the results showed the capacity of the recurrent neural network to predict growths carried out in different experimental conditions without using those used for its elaboration.     

Effect of stress induced by suboptimal growth factors on survival of Escherichia coli O157:H7.

This study investigated the growth and survival of E. coli O157:H7 exposed to a combination of suboptimal factors (22 degrees C, 7 degrees C, -18 degrees C/0.5% NaCl, 5.0% NaCl/pH 7.0, pH 5.4, pH 4.5/addition of lactic acid) in a simulation medium for red meat (beef gravy). Prolonged survival was noted as the imposed stress was more severe, and as multiple growth factors became suboptimal. At a defined temperature (7 degrees C or -18 degrees C), survival was prolonged at the more acid, more suboptimal pH (pH 4.5 > pH 5.4 > pH 7.0) while at a defined pH (pH 4.5), better survival was observed at 7 degrees C than at 22 degrees C. This suggests that application of the hurdle concept for preservation of food may inhibit outgrowth but induce prolonged survival of E. coli O157:H7 in minimal processed foods. At both 22 degrees C and 7 degrees C, the addition of lactic acid instead of HCl to reduce pH (to pH 4.5) resulted in a more rapid decrease of E. coli O157:H7. High survival was observed in beef gravy, pH 5.4 at -18 degrees C (simulation of frozen meat)-reduction of log 3.0 to log 1.9 after 43 days--and in beef gravy, pH 4.5 and 5% NaCl at 7 degrees C (simulation of a fermented dried meat product kept in refrigeration)--less than 1 log reduction in 43 days. In these circumstances, however, a high degree of sublethal damage of the bacterial cells was noted. The degree of sublethal damage can be estimated from the difference in recovery of the pathogen on the non-selective TSA medium and the selective SMAC medium.     

Effect of nisin on growth boundaries of Listeria monocytogenes Scott A, at various temperatures, pH and water activities

The effect of nisin on growth boundaries of Listeria monocytogenes Scott A in Tryptone Soy Broth (TSB) under different a(w)s, pH, and temperatures was studied. Growth/no growth turbidity data was modeled using logistic regression. Combinations of various temperatures (5-35 degrees C), pH (4.05-6.70) adjusted with HCl, a(w)s (0.937-0.998) NaCl (0.5-10.5%) and nisin (0-100 IU/ml) were used to monitor the growth/no growth response of L. monocytogenes Scott A for 60 days. The concordance of the logistic regression model was 99.4%, indicating successful data fitting. The minimum pH at which growth was observed was 4.81 at the temperature range of 25-35 degrees C and at a(w) as high as 0.992. Growth was observed at a(w) as low as 0.937, at pH 6.7, at the temperature range of 25-35 degrees C. Increasing nisin concentrations above 25 IU/ml resulted in a more inhibitory environment for L. monocytogenes. Presence of 100 IU/ml resulted in a minimum pH for growth at 5.20, and a minimum a(w) at 0.967 at the temperature range of 25-35 degrees C. It was remarkable that low to medium salt concentrations (2.5-4.5 NaCl% w/v) provided a protective effect against inhibition of L. monocytogenes by nisin. The present study points out the applicability of growth/no growth modeling in order to study any interactions between various factors affecting initiation of growth of micro-organisms, in which its turn helps the understudying of microbe-food ecosystem relations and the development of safer food.     

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.     

Combined effects of NaCl, NaOH, and biocides (monolaurin or lauric acid) on inactivation of Listeria monocytogenes and Pseudomonas spp

This study highlighted combinations of chemical stresses that could decrease or eliminate Listeria monocytogenes and Pseudomonas spp. surviving in food processing plants. Strains of L. monocytogenes, Pseudomonas fragi, and Pseudomonas fluorescens isolated from processing environments (meat and milk) were grown at 20 degrees C up to the early stationary phase. The strains were then subjected to 30 min of physicochemical treatments. These treatments included individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (fatty acids) challenges. Survival of the strains was studied after individual or combined acid (acetic acid), alkaline (NaOH), osmotic (NaCl), and biocides (monolaurin, lauric acid) challenges. Individual pH shocks had lower efficiencies than those used in combinations with other parameters. The treatment pH 5.4 followed by pH 10.5 had a low efficiency against L. monocytogenes. The opposite combination, pH 10.5 followed by pH 5.4, led to a 3-log reduction of the L. monocytogenes population. Pseudomonas spp. strains were much more sensitive than L. monocytogenes, and population reductions of 5 and 8 log (total destruction), respectively, were observed after the same treatments. As for L. monocytogenes, the combination pH 10.5 followed by pH 5.4 is more deleterious than the opposite. Whatever the bacterial species, the most efficient treatments were combinations of alkaline, osmotic, and biocide shocks. For instance, the combination pH 10.5 and 10% NaCl plus biocides showed reductions of 5 to 8 log for both bacteria. The origins of the observed lethal effects are discussed.     

Modeling the effect of storage atmosphere on growth-no growth interface of Listeria monocytogenes as a function of temperature, sodium lactate, sodium diacetate, and NaCl

The effect of aerobic and anaerobic conditions on growth initiation by a 10-strain composite of Listeria monocytogenes (10(4) CFU/ml) was evaluated in tryptic soy broth with 0.6% yeast extract (TSBYE) as a function of 220 combinations of pH (3.82 to 7.42), sodium lactate (SL) (0 to 10%, vol/vol), and sodium diacetate (SD) (0 to 0.5%, wt/vol) at 10 or 30 degrees C (a slightly abusive and the optimal growth temperature, both above the growth limiting range of 0 to 3 degrees C for L. monocytogenes) in 96-well microplates. In addition, four probability-of-growth models were developed to quantify the effect of 346 aerobic and 346 anaerobic combinations of temperature (4 to 30 degrees C), SL (0 to 6%, vol/vol), and SD (0 to 0.5%, wt/vol) in the presence of NaCl (0.5 or 2.5%, wt/vol) on the growth-no growth responses of the same L. monocytogenes strain composite, with a microplate reader. Growth responses were evaluated turbidimetrically (620 nm) every 5 days for a total of 40 days. Data were modeled with logistic regression to determine the growth-no growth interfaces. The minimum pH values at which growth of L. monocytogenes occurred were higher under anaerobic than under aerobic conditions, and this difference was more evident at 10 degrees C or at higher SL and SD concentrations. The MIC of SD decreased with increasing SL levels. Anaerobic storage reduced the levels of SL-SD, allowing the growth of L. monocytogenes compared with aerobic storage, especially at low temperatures. In the presence of 2.5% NaCl, the MICs for SD were lower than those obtained with 0.5% NaCl, especially at 4 and 10 degrees C, or in the presence of 5 to 6% SL. The developed models for anaerobic incubation showed good performance (80% successful predictions; i.e., in 40 of 50 comparisons) with independent data from studies on survival-growth of L. monocytogenes on meat products. The study provides quantitative data on the antimicrobial activity of SL (0 to 10%) and SD (0 to 0.5%), temperature (4 to 30 degrees C), and pH (3.82 to 7.42) and on the probability of growth of L. monocytogenes under anaerobic or aerobic conditions in the presence of 0.5 or 2.5% NaCl, and hence, addresses important needs for risk assessment activities.     

Effect of temperature and pH on survival of Listeria monocytogenes in coleslaw

The survival and growth of Listeria monocytogenes in fresh coleslaw, pH 3.9, and in coleslaw adjusted to pH 4.0, 5.0, 6.0 or 7.0 before inoculation was studied at three temperatures (4, 15 and 25 degrees C). L. monocytogenes was not detectable after 5 days incubation in fresh coleslaw nor in coleslaw adjusted to pH 4.0. Coleslaw at pH 5.0 was also inhibitory to L. monocytogenes at all three temperatures studied. A decline in viable numbers of L. monocytogenes in coleslaw at pH 6.0 occurred at 4 degrees C and at 15 degrees C, whereas at 25 degrees C the viable count of L. monocytogenes increased initially and remained high after incubation for 25 days. L. monocytogenes grew rapidly in coleslaw at pH 7.0 at all three temperatures studied, followed by an equally rapid decline in viable count.     

Growths kinetics comparison of clinical and seafood Listeria monocytogenes isolates in acid and osmotic environment

Comparison of pathogenic bacterial strains of clinical origin with strains of the same species isolated from the environment may be a valuable tool for microbial risk assessment, especially for foodborne pathogens. Thus, a number of Listeria monocytogenes strains responsible for human cases of listeriosis, in relation to the consumption of contaminated seafood, have been compared with "natural" L. monocytogenes strains isolated from similar seafood products. Complete factorial designs were used to assess quantitatively the growth abilities of four clinical and four seafood isolates of L. monocytogenes placed in various environmental conditions. The cells were submitted to acid and osmotic stress as they were in stationary phase (constant condition) or in exponential phase (dynamic condition). The effects and interactions of pH (5-7) and NaCl concentration (0.5-8% v/v) were studied at two growth temperatures (10 and 20 degrees C). Growth parameters (lag and generation times calculated with Gompertz equation) were used to compare the behavior of the strains with respect to the conditions of culture. The results indicated an overall weak effect of acid stress alone, whereas osmotic stress clearly affected bacterial growth and a synergic effect between these two factors was observed. Clinical strains displayed better adaptation than seafood strains in stationary phase, however, this difference was not verified in exponential phase. Low temperature (10 degrees C) usually confirmed the observations at 20 degrees C, and the differences between clinical and food strains were more pronounced. Finally, a classification of the eight strains, based on the collected data, showed three groups: (i) seafood strains, (ii) three clinical strains and (iii) the last clinical strain, alone due to its high resistance to adverse conditions.     

Heat and acid tolerance of Listeria monocytogenes after exposure to single and multiple sublethal stresses

The majority of published studies on the adaptive heat or acid tolerance response of Listeria monocytogenes have been performed with a single strain exposed to a single adaptation treatment; however, in food ecosystems, microorganisms commonly exist as multi-species communities and encounter multiple stresses, which may result in "stress hardening". Therefore, the present study evaluated the adaptive responses to heat (52, 57 and 63 degrees C) or lactic acid (pH 3.5) of a 10-strain composite of L. monocytogenes meat and human isolates at stationary phase, following exposure to combinations of osmotic (10% NaCl), acidic (pH 5.0 with HCl) and thermal (T; 46 degrees C) stresses, sequentially or simultaneously within 1.5h, in tryptic soy broth with 0.6% yeast extract (TSBYE). All treatments induced adaptive responses on L. monocytogenes at 57 degrees C, while no such cross-protection was observed at 52 and 63 degrees C. Survivor curves at 57 degrees C appeared convex with profound shoulders determined by a Weibull model. The highest thermotolerance was observed after combined exposure to acid and heat shock (pH-T), followed by exposure to osmotic shock, and by the combination of osmotic with heat shock (NaCl-T). Regarding acid tolerance, prior exposure to low pH, pH-T, or a combination of NaCl, pH and T resulted in a marked increase of resistance to pH 3.5, showing concave inactivation curves with tails at higher levels of survivors (log(10)CFU ml(-1)) than the control cultures. The sequence of exposure to sublethal stresses did not affect the thermotolerance of L. monocytogenes, whereas simultaneous exposure to most multiple stresses (e.g., NaCl-pH-T, NaCl-T and NaCl-pH) resulted in higher survivors of L. monocytogenes at pH 3.5 than exposure to the same stresses sequentially. The results indicate that combinations and sequences of sublethal hurdles may affect L. monocytogenes acid and heat tolerance, especially in acidic environments with mild heating or in low moisture environments.     

Effect of pH, NaCl content, and temperature on growth and survival of Arcobacter spp

Growth and survival of six human isolates of the pathogenic Arcobacter spp. in the presence of selected environmental factors were studied. Four strains of Arcobacter butzleri and two strains of Arcobacter cryaerophilus were exposed to pH levels of 3.5 to 8.0. Most strains grew between pH 5.5 and 8.0, with optimal growth of most A. butzleri and A. cryaerophilus strains at pH 6.0 to 7.0 and 7.0 to 7.5, respectively. The 24-h optimal growth range in the presence of NaCl was 0.5 to 1.0% for A. cryaerophilus. However, after 96 h, the optimum was between 0.5 and 2.0% NaCl. The optimum range for growth of A. butzleri strains was 0.09 to 0.5% NaCl after 96 h. The upper growth limits were 3.5 and 3.0% NaCl for A. butzleri and A. cryaerophilus, respectively. Survival at 25 degrees C in up to 5% NaCl was noted for A. butzleri 3556 and 3539 and A. cryaerophilus 3256. Decimal reduction times (D-values) at pH 7.3 in phosphate-buffered saline for three A. butzleri strains were 0.07 to 0.12 min at 60 degrees C, 0.38 to 0.76 min at 55 degrees C, and 5.12 to 5.81 min at 50 degrees C. At pH 5.5, decreased thermotolerance was observed, with D-values of 0.03 to 0.11 min at 60 degrees C, 0.30 to 0.42 min at 55 degrees C, and 1.97 to 4.42 min at 50 degrees C. Calculated z-values ranged from 5.20 to 6.28 degrees C. D-values of a three-strain mixture of A. butzleri in raw ground pork were 18.51 min at 50 degrees C and 2.18 min at 55 degrees C. Mild heat (50 degress C) followed by cold shock (4 or 8 degrees C exposure) had a synergistic lethal effect, reducing more cells than with an individual 50 degrees C treatment or with cold shock temperatures of 12 or 16 degrees C.     

Growth kinetics and cell morphology of Listeria monocytogenes Scott A as affected by temperature,NaCl, and EDTA

Growth kinetics and morphological characteristics of Listeria monocytogenes Scott A grown under stress conditions induced by increasing levels of NaCl and EDTA were studied as a function of temperature. L. monocytogenes Scott A was inoculated into brain heart infusion broth (pH 6) at 19, 28, 37, and 42 degrees C. Test cultures contained NaCl (at concentrations of 4.5, 6.0, and 7.5%) or EDTA (at concentrations of 0.1, 0.2, and 0.3 mM); control cultures contained 0.5% NaCl. Growth curves were fitted from plate count data by the Gompertz equation, and growth kinetics parameters were derived. Stationary-phase cells were examined by scanning and transmission electron microscopy. Generation times (GTs) and lag phase duration times (LPDs) increased as additive levels were increased. The bacterium grew at all NaCl levels. At 37 and 42 degrees C, growth was slow in media containing 7.5% NaCl, and no growth occurred in media containing 0.3 mM EDTA. Temperature was a major factor in certain stress conditions that led to cell elongation and loss of flagella. Cells in control media at 28 degrees C grew as short rods (0.5 by 1.0 to 2.0 microm), while at 42 degrees C most cells were 4 to 10 times as long. Higher levels of NaCl at higher temperatures resulted in longer and thicker cells. At 28 degrees C, 0.1 mM EDTA had little effect on growth kinetics and morphology; however, 0.3 mM EDTA caused a sixfold increase in GT and LPD and loss of flagellae, with most cells being two to six times as long as normal. Cell length did not correlate with growth kinetics. The results of this study suggest that the effect of altered morphological characteristics of L. monocytogenes cells grown under stress on the virulence and subsequent survival of these cells should be investigated.     

Survival and growth of Listeria monocytogenes in broth as a function of temperature, pH, and potassium lactate and sodium diacetate concentrations

The objective of this study was to determine the antimicrobial effect of a combination of potassium lactate and sodium diacetate (0, 1.8, 3, and 4.5%; PURASAL P Opti.Form 4, 60% solution) on the survival and growth of Listeria monocytogenes Scott A in pH-adjusted broth (5.5, 6.0, 6.5, and 7.0) stored at 4, 10, 17, 24, 30, and 37 degrees C. Appropriate dilutions of broth were enumerated by spiral plating on tryptose agar and counted with an automated colony counter. Growth data were iteratively fit, using nonlinear regression analysis to a three-phase linear model, using GraphPad PRISM. At pH 5.5, the combination of lactate-diacetate fully inhibited (P < 0.001) the growth of L. monocytogenes at all four levels and six temperatures. At pH 6.0, addition of 1.8% lactate-diacetate reduced (P < 0.001) the specific growth rate of L. monocytogenes and increased lag time; however, 3 and 4.5% completely inhibited the growth at the six temperatures studied. Efficacy of the lactate-diacetate mixture was decreased as pH increased and incubation temperature increased. Thus, at pH 6.5, at least 3% was required to retard (P < 0.001) the growth of L. monocytogenes in broth. There was a limited effect of the lactate-diacetate level on the specific growth rate of the pathogen at pH 7.0. However, 1.8 and 3% significantly lengthened the lag time at 4 and 10 degrees C. These results suggest that 1.8% of lactate-diacetate mixture can be used as a substantial hurdle to the growth of L. monocytogenes when refrigerated temperatures are maintained for products with pH less than 6.5.     

Survival and growth of alkali-stressed Listeria monocytogenes on beef frankfurters and thermotolerance in frankfurter exudates

Cells of Listeria monocytogenes exposed at 4 degrees C to 1% solutions of two alkaline cleaners or alkali-adapted in tryptose phosphate broth (pH 10.0) at 37 degrees C for 45 min, followed by 4 degrees C for 48 h, were inoculated onto beef frankfurters containing high fat (16 g) and high sodium (550 mg) or low fat (8 g) and low sodium (250 mg) per 57-g serving. Frankfurters were surface inoculated (2.0 log10 CFU/g), vacuum packaged, stored at -20, 4, or 12 degrees C, and analyzed for populations of L. monocytogenes at 2-day to 2-week intervals. Populations did not change significantly on frankfurters stored at -20 degrees C for up to 12 weeks. After storage at 4 degrees C for 6 weeks (I week before the end of shelf life), populations of control cells and cells exposed to alkaline cleaners were ca. 6.0 log10 CFU/g of low fat, low sodium (LFLS) frankfurters and ca. 3.5 log10 CFU/g of high fat, high sodium (HFHS) frankfurters. Growth of alkali-adapted cells on both types of frankfurters was retarded at 4 degrees C. Growth of L. monocytogenes on frankfurters stored at 12 degrees C was more rapid than at 4 degrees C, but a delay in growth of alkali-adapted cells on HFHS and LFLS frankfurters was evident during the first 9 and 6 days, respectively. Alkali-adapted cells had a significantly (P < or = 0.05) lower logistic D59 degrees C-value (decimal reduction time) than alkaline cleaner-exposed cells, but the D59 degrees C-value was not different from that of control cells. Cells exposed to a nonbutyl alkaline cleaner, and then heated in LFLS frankfurter exudates, had a significantly lower D62 degrees C-value than cells that had been exposed to some of the other treatments. Growth characteristics of L. monocytogenes inoculated onto the surface of frankfurters may be altered by previous exposure to alkaline environments. Differences in growth characteristics of L. monocytogenes on HFHS versus LFLS beef frankfurters stored at refrigeration temperatures indicate that composition influences the behavior of both alkaline-stressed and control cells.     

Survival characteristics and the applicability of predictive mathematical modelling to Listeria monocytogenes growth in sous vide products

Survival and growth of Listeria monocytogenes isolates during sous vide processing and storage, and the applicability of predictive modelling in determining the potential for growth of L. monocytogenes in broth models and in sous vide products was investigated. L. monocytogenes grew in anaerobic tryptose phosphate broth and in chicken and beef samples by 2 log cycles in 8 days at 3 degrees C and 4-5 log cycles in 6 days at 8 degrees C. However, heating to an internal temperature of 70 degrees C resulted in a 4-5 log reduction and 70 degrees C/2 min resulted in a reduction greater than 7 log cycles. Lowering the product pH to 5.0 was effective in inhibiting L. monocytogenes growth, whereas a sodium chloride concentration of 2% had a negligible effect on growth rates. The square root model (Ratkowsky et al., 1983) predicted L. monocytogenes growth rates at 0-25 degrees C with a coefficient of determination (R2 value) of 98.36-99.63% and a bias factor of 1.08 to 1.21 in beef, chicken and broth substrates of unmodified pH. In addition, the Response Surface Polynomial Model (Version 3.1, Buchanan et al., 1989) predicted generation times at 5-25 degrees C with a 0-17.4% difference between observed and expected generation times in tryptose phosphate broth at pH 7.3. There were however, large differences (25.5 vs. 5.3 h) between observed generation times at pH 5.6 (8 degrees C) and those predicted by the Pathogen Modelling Program in tryptose phosphate broth. A divergence from predicted values was also noted at lower temperatures (0-3.5 degrees C) in the square root model.     

Impact of preheating on the behavior of Listeria monocytogenes in a broth that mimics Camembert cheese composition

The effect of preheating on the survival of L. monocytogenes in Richard's broth, which mimics the composition of Camembert cheese composition, was examined. Experiments were carried out to reproduce contamination of cheese with environmental heat-stressed cells of L. monocytogenes surviving hot-cleaning procedures. Cells in mid-log phase were heated for 30 min at 56 degrees C before being inoculated into Richard's broth. The pHs and temperatures of Richard's broth were chosen to recreate the conditions of curd dripping (pH 5, 25 degrees C), of the beginning of cheese ripening (pH 5, 12 degrees C), and of the beginning (pH 5, 4 degrees C) and the end (pH 7, 4 degrees C) of cheese storage. Immediately after heat treatment, the viability loss was especially high for strain 306715, which exhibited only 0.6% +/- 0.2% survival, compared with 22% +/- 8.7% for strain EGD. The percentages of the surviving heated cells that were injured were 93% +/- 8% for strain 306715 and 98% +/- 3% for strain EGD. The destruction of the surviving L. monocytogenes cells was accelerated when they encountered the pH and temperature conditions of Camembert cheese during manufacturing, ripening, and cold storage (pH 5 at 25, 12, and 4 degrees C, respectively). The multiplication of the surviving heated cells was retarded under favorable growth conditions similar to those of storage by the distributor and the consumer (pH 7 at 4 and 12 degrees C, respectively).     

Acid and NaCl limits to growth of Listeria monocytogenes and influence of sequence of inimical acid and NaCl levels on inactivation kinetics

Variability in growth limits of Listeria monocytogenes in response to low pH (adjusted with HCl) or high salinity (NaCl) was evaluated for 127 strains in brain heart infusion broth at 25 degrees C. Over 95% of strains habituated at pH 5.0 grew subsequently at pH 4.2, while 25% were able to grow at pH 4.1. More than 85% of strains preadapted to growth at 8.5% NaCl (wt/vol) subsequently grew in the presence of 11.3% NaCl, while 25% were able to grow at 13% NaCl, and 4.7% grew in the presence of 13.9% NaCl. The results extend the generally accepted growth limits for L. monocytogenes in response to these hurdles. Two strains, one of which was relatively tolerant of both hurdles, and another that was less tolerant of both hurdles, were subjected to different sequences of lethal acid (pH 3.5) and NaCl (14%, wt/vol) stresses to determine whether survival was affected by growth limits, or by sequence of application of treatment. There was no significant difference in the inactivation kinetics of the two strains, but inactivation rates were affected by different treatments. For both strains, the inactivation rates, from fastest to slowest, resulted from: (i) lethal pH and then by lethal water activity, or lethal water activity and then by lethal pH; (ii) lethal pH and water activity applied simultaneously; (iii) lethal pH; and (iv) lethal water activity. The results demonstrated that the sequence of lethal stress application strongly influences L. monocytogenes inactivation, and that L. monocytogenes growth limits are not good predictors of survival in inimical environments.     

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.     

The influence of mayonnaise pH and storage temperature on the growth of Listeria monocytogenes in seafood salad

Seafood salad has been identified as a ready-to-eat food with a relatively high incidence of contamination by Listeria monocytogenes; however, little is known about the behavior of this pathogen in seafood salad as a function of product pH and storage temperature. To produce data towards the development of a predictive growth model, a 6-strain cocktail of L. monocytogenes was inoculated onto the surface of a shrimp-crabmeat product, mixed with mayonnaise that was previously adjusted with NaOH to pH 3.7, 4.0, 4.4, 4.7 or 5.1, and then stored at 4 degrees , 8 degrees or 12 degrees C under both aerobic and vacuum conditions. At each storage temperature, L. monocytogenes was able to grow in the seafood salad under both aerobic and vacuum conditions. The slowest growth of L. monocytogenes was observed in seafood salad with a mayonnaise pH of 3.7 and a storage temperature of 4 degrees C under vacuum condition. In salad with the same mayonnaise pH, the growth rate (GR, log10 cfu/h) of L. monocytogenes increased as a function of storage temperature. At the same storage temperature, the lag phase duration (LPD, h) of L. monocytogenes decreased as mayonnaise pH increased. At the same mayonnaise pH and temperature, LPD of L. monocytogenes was greater under aerobic than under vacuum conditions. Regression analyses indicated that mayonnaise pH is the main effector on the LPD of L. monocytogenes in seafood salad, and storage temperature was the main effector on the GR. Secondary models that describe LPD and GR of L. monocytogenes in seafood salad as a function of mayonnaise pH and storage temperature were produced.     

Predictive thermal inactivation model for Listeria monocytogenes with temperature, pH, NaCl, and sodium pyrophosphate as controlling factors.

The effects and interactions of heating temperature (55 to 65 degrees C), pH (4 to 8), salt (NaCl; 0 to 6%, wt/vol), and sodium pyrophosphate (SPP; 0 to 0.3%, wt/vol) on the heat inactivation of a four-strain mixture of Listeria monocytogenes in beef gravy were examined. A factorial experimental design comparing 48 combinations of heating temperature, salt concentration, pH value, and SPP content was used. Heating was carried out using a submerged-coil heating apparatus. The recovery medium was plate count agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve-fitting program. The D-values were analyzed by second-order response surface regression for temperature, pH, NaCl, and SPP levels. Whereas increasing the NaCl concentration protected L. monocytogenes against the lethal effect of heat, high SPP concentrations increased heat sensitivity. Also, low pH values increased heat sensitivity of L. monocytogenes. The four variables interacted to affect the inactivation of the pathogen. Thermal resistance of L. monocytogenes can be lowered by combining these intrinsic factors. A predictive model that described the combined effect of temperature, pH, NaCl, and SPP levels on thermal resistance of L. monocytogenes was developed. The model can predict D-values for any combination of temperature, pH, NaCl, and SPP that are within the range of those tested. Using this predictive model, food processors should be able to design adequate thermal regimes to eliminate L. monocytogenes in thermally processed foods.     

Survival,growth, and inactivation of acid-stressed Shigella flexneri as affected by pH and temperature

A study was done to determine the survival, growth, and inactivation characteristics of unadapted, acid-adapted, and acid-shocked Shigella flexneri 2a cells as affected by pH and temperature. The pathogen was grown at 37 degrees C for 18 h in tryptic soy broth containing no glucose (TSBNG) (unadapted cells) and TSBNG supplemented with 1% glucose (TSBG) (acid-adapted cells). Cells grown in TSBNG were acid-shocked by adjusting 18-h cultures to pH 4.5+/-0.05 with lactic acid. All three cell types were separately inoculated into tryptic soy broth (6.6-7.0 log(10) cfu/ml) containing 0.25% glucose (TSB) acidified to pH 3.5-5.5 with lactic acid and incubated at 4, 12, 21, 30, and 48 degrees C for up to 144 h. Overall, inactivation of S. flexneri cells at low pH was enhanced with an increase in incubation temperature. All three types of cells survived for 144 h at 4 degrees C in TSB acidified to pH 3.5, compared to < 24 h at 30 degrees C and 2 h at 48 degrees C. The population of all three cell types increased significantly (alpha = 0.05) within 24 h when cells were incubated at 12, 21, or 30 degrees C in TSB at pH 5.0, 5.5, or 7.3. Prior exposure of the S. flexneri to an acidic environment (acid-adapted or acid-shocked cells) resulted in increased resistance to extreme acid and temperature conditions. Acid-adapted cells decreased by approximately 2.5 log(10) cfu/ml when incubated at 4 degrees C for 144 h, compared to a 6-log(10) reduction in control (unadapted) cells. When cells were exposed to low pH (3.5-4.5) and high temperature (48 degrees C), significantly higher (alpha = 0.05) populations were recovered on tryptic soy agar (TSA) than on TSA supplemented with 4% NaCl (TSAS), indicating that a portion of S. flexneri cells were injured. Results show that the ability of S. flexneri to survive and grow at a given pH is influenced by previous exposure to acidic environments and by incubation temperature.