The effect of temperature and low pH on survival of Shigella flexneri in broth

The survival characteristics of Shigella fiexneri strain 5348 were determined in brain heart infusion broth as a function of low pH (2 to 5) and temperature (4 to 37 degrees C). Stationary-phase cells were inoculated into sterile media to give initial populations of 6 to 7 log10 CFU/ml. Bacterial populations were determined periodically by aerobic plate counts. Survivor curves were fitted from plate count data using a two-phase linear model to derive lag times and slopes of the curves, from which D-values and times to a 4-D (99.99%) inactivation (T4D) were calculated. In general, survival increased as temperature decreased and as pH increased. Bacterial populations reached undetectable levels (<1.3 log10 CFU/ml) at 37, 28, 19, 12, and 4 degrees C in media adjusted to pH 4 after 5, 15, 23, 85, and 85 days, respectively, and in media adjusted to pH 3 after 1, 7, 9, 16, and 29 days, respectively. In media adjusted to pH 2, bacterial populations were stable for 2 to 12 h at temperatures of 19 degrees C or lower and reached undetectable levels after 1 to 3 days, while at 28 and 37 degrees C, the bacteria were undetectable after 8 and 2 h, respectively. In media adjusted to pH 5, bacterial levels decreased only 0.5 to 1.5 log10 CFU/ml after 75 days at 4 degrees C and decreased to undetectable levels after 135 days at 12 degrees C, while growth occurred at higher temperatures. These results indicate that S. flexneri is acid resistant and that acidic foods may serve as vehicles for infection.     

Effect of organic acids and temperature on survival of Shigella flexneri in broth at pH 4

The survival of bacterial pathogens in acidified foods depends not only on the hydrogen ion concentration, but also on the type of acid and the storage temperature. Shigella flexneri is a foodborne pathogen that is acid tolerant. The survival of S. flexneri 5348 in brain heart infusion broth supplemented with 0.04 M acetic, citric, lactic, malic, or tartaric acid and adjusted to pH 4 with HCI or NaOH was studied. The control medium was brain heart infusion broth adjusted to pH 4 with HCI. Stationary-phase cells were inoculated into media at initial populations of 6 to 7 log10 CFU/ml and incubated at 4, 19, 28, and 37 degrees C. A two-phase linear inactivation model was applied to plate count data to derive lag times (tL) and slopes of the curves, from which D-values and time required for a 4-log10 decrease in population (T4D) were calculated. In all cases, survival increased with decreasing temperature. For each acid, tL, the D-value, and T4D increased with decreasing temperature. All acids inhibited S. flexneri to some extent but to differing degrees as follows: lactic acid, acetic acid > citric acid, malic acid, tartaric acid > HCl. The T4D values for the control medium and for media containing acetic, citric, lactic, malic, and tartaric acids were 64, 47, 50, 34, 58, and 52 h, respectively, at 37 degrees C and 2,607, 1,498, 1,905, 1,346, 1,726, and 2,134 h, respectively, at 4 degrees C. The results of this study indicate that organic acids may aid in the inactivation of Shigella. However, these data also suggest that foods stored at or below room temperature containing low levels (< 1%) of acids could cause illness if contaminated with Shigella.     

The effect of NaCl on survival of Shigella flexneri in broth as affected by temperature and pH

Shigella, a major foodborne pathogen, survives well in salt-containing environments. However, systematic data are scarce. We studied the behavior of Shigella flexneri 5348 in brain heart infusion broth (pH 4 to 6) containing 0.5 to 8% NaCl. Stationary-phase cells were inoculated into sterile media at initial concentrations of 6 to 7 log10 CFU/ml and incubated at 12 to 37 degrees C. Bacterial population sizes were determined periodically by plate counts. Survivor curves were derived from plate count data by using a two-phase linear model to determine lag times and slopes of the curves, from which decimal reduction times (D-values) and times to a 4-log10 inactivation (T4D) were calculated. In media of pH 6, the bacteria grew in the presence of < or = 6% NaCl at 19 and 37 degrees C and in the presence of < or = 7% NaCl at 28 degrees C. In media of pH 5, growth was observed in the presence of < or = 2, < or = 4, < or = 4, and 0.5% NaCl at 37, 28, 19, and 12 degrees C, respectively. Growth did not occur and bacterial populations gradually declined in media of pH 4. While NaCl had a major effect on growth, bacterial survival was affected to a lesser extent. Lag times decreased with increasing NaCl levels; however, the effect on D-values and T4D values was less pronounced. The average T4D values for media of pH 4 containing 0.5 to 6% NaCl were 4, 13, 23, and 61 days at 37, 28, 19, and 12 degrees C, respectively. These results show that S. flexneri is salt tolerant and suggest that salty foods may serve as vehicles for infection with this bacterium.     

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.     

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.     

Modeling the combined effects of pH, temperature and ascorbic acid concentration on the heat resistance of Alicyclobacillus acidoterrestis

In this study, thermal inactivation parameters (D- and z-values) of Alicyclobacillus acidoterrestris spores in McIlvaine buffers at different pH, apple juice and apple nectar produced with and without ascorbic acid addition were determined. The effects of pH, temperature and ascorbic acid concentration on D-values of A. acidoterrestris spores were also investigated using response surface methodology. A second order polynomial equation was used to describe the relationship between pH, temperature, ascorbic acid concentration and the D-values of A. acidoterrestris spores. Temperature was the most important factor on D-values, and its effect was three times higher than those of pH. Although the statistically significant, heat resistance of A. acidoterrestris spores was not so influenced from the ascorbic acid within the concentration studied. D-values in apple juice and apple nectars were higher than those in buffers as heating medium at similar pH. The D-values ranged from 11.1 (90 °C) to 0.7 min (100 °C) in apple juice, 14.1 (90 °C) to 1.0 min (100 °C) in apple nectar produced with ascorbic acid addition, and 14.4 (90 °C) to 1.2 min (100 °C) in apple nectar produced without ascorbic acid addition. However, no significant difference in z-values was observed among spores in the juices and buffers at different pH, and it was between 8.2 and 9.2 °C. The results indicated that the spores of A. acidoterrestris may survive in fruit juices and nectars after pasteurization treatment commonly applied in the food industry.     

Use of an autobioluminescent Campylobacter jejuni to monitor cell survival as a function of temperature, pH, and sodium chloride

The effects of storage temperature (4, 22, and 30 degrees C), pH (4.0 to 8.5), and sodium chloride concentration (0.25 to 7.5%, wt/vol) on the survival of a strain of Campylobacter jejuni ATCC 35921 with an autobioluminescent phenotype (generated from the fusion of luxCDABE genes from Xenorhabdus luminescens to the C. jejuni flaA promoter) in Mueller-Hinton broth under aerobic conditions were determined. At comparable pHs, autobioluminescent C. jejuni cells die most rapidly at 30 degrees C and most slowly at 4 degrees C. For example, at pH 5.5, a 1.2-log decrease in the cell count was observed after 4 days of storage at 4 degrees C, while 3.7- and 4.8-log reductions in cell numbers were observed after 4 days of storage at 22 and 30 degrees C, respectively. At 4 degrees C, C. jejuni was sensitive to NaCl concentrations of > or = 2.5%. However, the degree of inactivation at this storage temperature was also significantly lower than that observed at 22 degrees C, which, in turn, was substantially lower than that observed at 30 degrees C. In the presence of different NaCl concentrations at 22 and 30 degrees C, a strong correlation (r = 0.98) between plate count and bioluminescence output was obtained. However, at low pHs (4.0 and 4.5) and/or a low storage temperature (4 degrees C), bioluminescence did not correlate well with plate count. Despite these limitations, an autobioluminescent phenotype of C. jejuni proved to be a useful tool for studying the behavior of C. jejuni exposed to environmental stresses.     

Preparation and validation of a growth model for Bacillus cereus: the effects of temperature, pH, sodium chloride and carbon dioxide

The growth responses of a vegetative inoculum of Bacillus cereus as influenced by varying conditions of temperature, pH value and sodium chloride concentration (% w/v) and carbon dioxide concentration (% v/v) were determined in laboratory medium. Growth curves in concentrations of NaCl in the range 0.5–10.5% (w/v), pH values in the range 4.5–7.0, CO₂ concentrations in the range 10–80% (v/v) and storage temperatures from 10 °C to 30 °C were fitted using the regime of Baranyi et al. (1993). A response surface model was prepared and predictions of doubling time, growth rate, lag time and time to 1000-fold increase could be obtained for any set of conditions within the matrix studied. This model is included in Food MicroModel Version 1. Predicted doubling times from the model were compared to observed doubling times in the literature and the model was found to give realistic estimates of doubling time for a range of foods including milk, meat and poultry and carbohydrate-based products.     

Modeling the growth kinetics of Bacillus cereus as a function of temperature, pH, sodium lactate and sodium chloride concentrations

Mathematical models describing the growth kinetic parameters (lag phase duration and growth rate) of Bacillus cereus as a function of temperature, pH, sodium lactate and sodium chloride concentrations were obtained in this study. In order to get a residual distribution closer to a normal distribution, the natural logarithm of the growth kinetic parameters were used in modeling. For reasons of parsimony, the polynomial models were reduced to contain only the coefficients significant at a level of p相似文献   

Effect of sodium chloride concentration on pH, water-holding capacity and extractable protein of prerigor and postrigor ground beef

Fifteen beef cattle of similar age and management history were randomly allotted by slaughter days into three groups. Paired sternomandibularis were removed immediately following bleeding and trimmed of visible fat and connective tissue. They were randomly labelled as prerigor and postrigor and assigned to a 0, 0·5, 1·0, 2·0 or 4·0% NcCl treatment. Water-holding capacity (WHC), pH, the ratio of absorbance at 250 nm over the absorbance at 260 nm (R-values), and 1·0m NaCl extractable protein (EP) were monitored over treatment times. The 0 h samples were defined as when the NaCl was incorporated with the muscle. R-values verified that 0 h samples were in the prerigor or postrigor state. Ultimate pH remained higher (P < 0·05) in prerigor homogenates with increasing NaCl concentration. EP and WHC were higher (P < 0·05) in prerigor than in postrigor homogenates with 2 and 4% NaCl at all time periods. Prerigor homogenates containing 0·5 and 1·0% NaCl had higher (P < 0·05) WHC at 12, 24, 24, 48 and 96h than similarly treated postrigor homogenates and as high or higher WHC than any postrigor treatment. Results of this study indicate an advantage to using low NaCl concentrations in prerigor salted beef.     

The effect of pH,sodium chloride,sodium nitrite and storage temperature on the growth of Clostridium perfringens and faecal streptococci in laboratory media

The effects of salt concentration (1–12% w/v) in combination with unheated sodium nitrite (0–400 μg/ml) on growth of mixed strains of Clostridium perfringens and of faecal streptococci at three pH values (5.6, 6.2, 6.8) and storage temperatures ranging from 10°C to 35°C is reported. At pH 6.2, following storage at 15°C, 1% salt and 50 μg/ml nitrite inhibited growth of C. perfringens. At 20°C and pH 6.2, 200 μg/ml nitrite plus 3% salt, or 50 μg/ml plus 4% salt were required to inhibit growth. Growth of C. perfringens was prevented by levels of curing salts used commercially providing the pH was 6.2 or below. At pH 6.8 or above at least 4% salt and 50 μg/ml nitrite was required to prevent growth at 20°C. The faecal streptococci grew in medium containing 6% salt and 400 μg/ml nitrite irrespective of pH or storage temperature. In 8% salt growth was prevented by storing at or below 17.5°C or, if pH was 6.2 or lower, by adding 200 μg/ml nitrite irrespective of storage temperature. Growth of faecal streptococci was not controlled by concentrations of curing salts which would be acceptable in meat products.     

Growth kinetics of Vibrio parahaemolyticus O3:K6 under varying conditions of pH, NaCl concentration and temperature

The growth responses of Vibrio parahamolyticus to pH, NaCl concentration and temperature changes were studied using serotype O3:K6 and other strains. Growth curves were obtained for 27 different sets of conditions, comprised of three levels of NaCl concentration, pH and temperature. The temperature, pH and NaCl concentrations most favorable for growth were in the order of 25 degrees C, 20 degrees C and 15 degrees C, pH 8, 7 and 5.8, and 1%, 3% and 7%, respectively. The bacteria grew most rapidly at 25 degrees C, at a pH of 7 or 8 in the presence of 1% or 3% NaCl, with the population (initial, ca. 2.5 log CFU/mL) reaching a level log 7 CFU/mL at 12 h. A growth predictive model using the Gompertz equation was generated from the experimental data for any combination of NaCl concentration, pH and temperature within the range used in this study.     

The effect of phosphate, sodium chloride, sodium nitrite, storage temperature and pH on the growth of enteropathogenic Escherichia coli in a laboratory medium

The effect of eight phosphates in combination with sodium chloride/nitrite mixtures on growth of mixed strains of enteropathogenic Escherichia coli over a period of 10 weeks at three pH values (5.6, 6.2, 6.8) and at seven temperatures ranging from 10 degrees C to 35 degrees C is reported. All eight phosphates inhibited growth to varying degrees in at least some of the conditions investigated. Instances of inhibition increased with concentration of sodium chloride/nitrite and were more frequent at lower temperatures and pH values.     

The impact of lactic acid concentration and sodium chloride on pH, water-holding capacity, and cooked color of injection-enhanced dark-cutting beef

The objectives of this study were to evaluate the effect of enhancement with 0.5%, 1.0%, 1.5%, or 2.0% lactic acid (LA), with or without 0.5% salt (NaCl), on pH, water-holding capacity, and resultant cooked color of dark-cutting (DC) beef. The pH of DC sections treated with LA decreased (P < 0.05), whereas water-holding capacity (WHC) increased (P < 0.05) with increasing concentrations of LA. Sensory panelists noted an increase (P < 0.05) in the degree of doneness (less pink internal color), whereas, a^* and chroma values were lower (P < 0.05), and hue angles were greater (P < 0.05), indicating the internal color of LA-treated DC sections was less red and appeared more well-done. Also, 630:580 nm reflectance ratios were similar (P > 0.05) among DC steaks treated with 0.5% and 1.0% LA and normal pH (NDC) steaks, likely caused by an increase (P < 0.05) in myoglobin denaturation in LA-enhanced DC steaks. These results indicate that the use of LA can reduce postmortem muscle pH and alter the cooked color of DC beef, ultimately resulting in a deletion of the persistent pinking condition.     

Quantifying the effects of heating temperature, and combined effects of heating medium pH and recovery medium pH on the heat resistance of Salmonella typhimurium

The influence of heating treatment temperature, pH of heating and recovery medium on the survival kinetics of Salmonella typhimurium ATCC 13311 is studied and quantified. From each non-log linear survival curve, Weibull model parameters were estimated. An average shape parameter value of 1.67 was found, which is characteristic of downward concavity curves and is in agreement with values estimated from other S. typhimurium strains. Bigelow type models quantifying the heating temperature, heating and recovery medium pH influences are fitted on scale parameter delta data (time of first decimal reduction), which reflects the bacterial heat resistance. The estimate of z(T) (4.64 degrees C) is in the range of values given in the literature for this species. The influence of pH of the heating medium on the scale parameter (z(pH): 8.25) is lower than that of the recovery pH medium influence (z(')(pH): 3.65).     

Modelling Salmonella concentration throughout the pork supply chain by considering growth and survival in fluctuating conditions of temperature, pH and a(w)

We aim to predict the population density of Salmonella spp. through the pork supply chain under dynamic environmental conditions (pH, a(w) and temperature) that fluctuate from growth to survival/slow inactivation. To do this, the dependence of the probability of growth, and of the growth and inactivation rate on the temperature, pH and a(w) were modelled. Probabilistic and kinetic measurements, i.e. growth and survival curves, were collected from the ComBase database (www.combase.cc). Conditions at which selected data used to fit the models were generated covered wide ranges that are relevant to the pork supply chain. Probabilistic and kinetic models were combined to give predictions on the concentration of Salmonella spp. at any stage of the pork supply chain under fluctuating pH, a(w) and/or temperature. Models were implemented in a user-friendly computing tool freely available from http://www.ifr.ac.uk/safety/SalmonellaPredictions/. This program provides estimates on the population dynamics of Salmonella spp. at any stage of the pork supply chain and its predictive performance has been validated in several pork products.     

Modeling the germination kinetics of clostridium botulinum 56A spores as affected by temperature, pH, and sodium chloride

The germination kinetics of proteolytic Clostridium botulinum 56A spores were modeled as a function of temperature (15, 22, 30 degrees C), pH (5.5, 6.0, 6.5), and sodium chloride (0.5, 2.0, 4.0%). Germination in brain heart infusion (BHI) broth was followed with phase-contrast microscopy. Data collected were used to develop the mathematical models. The germination kinetics expressed as cumulated fraction of germinated spores over time at each environmental condition were best described by an exponential distribution. Quadratic polynomial models were developed by regression analysis to describe the exponential parameter (time to 63% germination) (r2 = 0.982) and the germination extent (r2 = 0.867) as a function of temperature, pH, and sodium chloride. Validation experiments in BHI broth (pH: 5.75, 6.25; NaCl: 1.0, 3.0%; temperature: 18, 26 degrees C) confirmed that the model's predictions were within an acceptable range compared to the experimental results and were fail-safe in most cases.     

Proteolytic activity of isolated lamb calpains on myofibrils under the conditions of pH, Ca2+ concentration and temperature existing in postmortem muscle.

The two calcium-activated neutral proteinases (calpains I and II) and their specific inhibitor were isolated by ion exchange chromatography in DEAE-Sephacel from lamb skeletal muscle (longissimus dorsi). Their proteolytic activities were then determined using myofibrils as substrate. The Ca2+ requirements were different for each form of the enzyme: calpain I needed only 50 mumol Ca2+ for half-maximal activity, while the other isoenzyme, calpain II, needed 1,000 mumol Ca2+ for reaching 50% of its maximum activity. Both calpains showed a relevant activity in the pH range 5.5-6.5 (over 40% of maximum activity found at pH 7.5). With regard to the effect of temperature, both isoenzymes retained about 25% of their activity at 25 degrees C with a temperature reduction down to 4 degrees C. It is concluded that calpain I is an active protease under conditions similar to that prevalent in lamb meat during postmortem storage.     

The combined effects of fat content,calcium chloride,and coagulant concentration on the development of cheese curd structure

The effects of differing content levels of calcium chloride (approximately 200 and 400 μg Ca per 100 g milk protein) and of a microbial coagulant (3200 and 6400 μL per 100 L of milk; 950 IMCU (international milk coagulating units) mL⁻¹) on the coagulation of cows' milk with various fat levels (0.02–3.77%, w/w) was studied. Non-linear regression analysis was used to evaluate dynamic factors (lag time, t_lag; maximum coagulation rate, C_max; time for the maximum coagulation rate, t_max). Increasing fat content in the milk at constant calcium chloride and coagulant contents had no significant and clear effects on the t_lag, C_max, and t_max values. Increased levels of calcium chloride or microbial coagulant led to a significant decrease in t_lag and t_max, and conversely increased C_max. Therefore, milk fat content had no significant effect on gel development; however, levels of calcium chloride and coagulant significantly influenced gel structure.