Temperature influence on Penicillium citrinum Thom growth and citrinin accumulation kinetics

To study the temperature influence on both Penicillium citrinum growth and citrinin accumulation, a strain isolated from corn was cultured on Czapek agar with maize extract at 15, 20, 25 and 30 °C. Radial growth rate and lag phase were determined from the increase in colony diameter with time. The optimal temperature for P. citrinum growth was 30°C. Citrinin extracted from the agar medium was determined by thin layer chromatography. Citrinin accumulation kinetics were analyzed by fitting the data to curves generated by using a logistic function. The parameters obtained from this equation demonstrated, for all temperatures studied, that the maximum citrinin accumulation by P. citrinum on Czapek agar with maize extract was at about 30°C. At 37°C a rapid decrease in the citrinin concentration was observed after a maximal value was reached.     

Development of twoListeria monocytogenesgrowth models in a meat broth and their application to beef meat

Growth variation ofListeriastrains was taken into account by building two growth models with strains previously characterized, respectively, by their slow (L. monocytogenes CLIP 19532) and fast (L. monocytogenes 14) growth in different conditions of pH, a_wand temperature. Strains of intermediate growth were studied in a meat broth and strains used for the models were grown on the surface of beef meat. Ten growth repetitions at 14°C– a_w0.98–pH 6.2 showed that generation times were similar [ratio value (R=standard deviation/average): 3.2%] but that range of lag times was wide (R=27.4%). In broth, calculated lag and generation times were not significantly different between strains from 30 to 14°C, but variations became larger as temperatures came close to 4°C. Model values corresponded well to experimental generation times and to a lesser extent to lag times. On meat at 4°C and 14°C both strains had experimental lag times three-fold longer than predicted lag times. Experimental generation times were shorter than predicted values at 14°C and longer at 4°C: differences between growth in broth and on meat could be due to the characteristics of the meat, the experimental conditions of growth, the mode of inoculation and the way of adjustment of a_w. Growth variations were found between available predictive models.     

Short communication: Comparison of growth kinetics at different temperatures of Streptococcus macedonicus and Streptococcus thermophilus strains of dairy origin

Within the genus Streptococcus, S. thermophilus and S. macedonicus are the 2 known species related to foods. Streptococci are widely used as starter cultures to rapidly lower milk pH. As S. macedonicus has been introduced quite recently, much less information is available on its technological potential. Because temperature is an important factor in fermented food production, we compared the growth kinetics over 24 h of 8 S. thermophilus and 7 S. macedonicus strains isolated from various dairy environments in Italy, at 4 temperatures, 30°C, 34°C, 37°C and 42°C. We used the Gompertz model to estimate the 3 main growth parameters; namely, lag phase duration (λ), maximum growth rate (µ_max), and maximum cell number at the stationary phase (N_max). Our results showed significant differences in average growth kinetics between the 2 species. Among the strains tested, 37°C appeared to be the optimal temperature for the growth of both species, particularly for S. macedonicus strains, which showed mean shorter lag phases and higher cell numbers compared with S. thermophilus. Overall, the growth curves of S. macedonicus strains were more similar to each other whereas S. thermophilus strains grew very differently. These results help to better define and compare technological characteristics of the 2 species, in view of the potential use of S. macedonicus in place of S. thermophilus in selected technological applications.     

Induction of a lag phase by chiller temperatures in Escherichia coli growing in broth or on pork

When log phase cultures of Escherichia coli in brain–heart infusion (BHI) were cooled from 12°C to temperatures below 7°C (the minimum required for growth), the optical densities of cultures increased at declining rates for up to 6 days during incubation at temperatures between 6 and 3°C inclusive, but did not increase at temperatures ≤2°C. However, the numbers of E. coli recovered from cultures on selective or non-selective agars were similar at all times of incubation up to 8 days at temperatures below the minimum for growth. From optical density measurements it appeared that when log phase cultures were cooled to 2°C, then returned to 15°C, a lag developed with time at 2°C to reach a maximum of about 1 h after about 4 h. When cultures were returned to 12°C after times at 2°C between 0·5 and 8 days, the time during which optical densities did not increase was constant at about 2 h, but the initial rate of optical density increase declined with time at 2°C. On pork fat tissue the lag time, determined by increases in the numbers of E. coli cfu on tissues inoculated with cells returned from incubation at 2°C for 16 h to a growth permitting temperature between 7 and 30°C, was longer than the lag time determined from optical density measurements of broth cultures subjected to the same temperature regime. Lag times for E. coli adapted to and growing on normal pH pork lean tissue were longer than for E. coli on fat tissue, while unadapted E. coli did not initiate growth on lean tissue, after incubation at 2°C, at 12°C or lower temperatures. The observations indicate that lag phase development in log phase E. coli subjected to chiller temperatures is complex, and that prediction of lag resolution in log phase cells exposed to temperatures that fluctuate around the minimum for growth will require modeling of lag induction as well as lag resolution.     

The growth and aflatoxin production of Aspergillus flavus strains on a cheese model system are influenced by physicochemical factors

Traditional cheeses may be contaminated by aflatoxin-producing Aspergillus flavus during the ripening process, which has not been sufficiently taken into account. The objectives of this study were to evaluate the influence of water activity (a_w), pH, and temperature on the lag phases, growth, and aflatoxin production of 3 A. flavus strains (CQ7, CQ8, and CG103) on a cheese-based medium. The results showed that the behavior of A. flavus strains was influenced by pH, a_w, and temperature conditions. The CQ7 strain showed the maximum growth at pH 5.5, 0.99 a_w, and 25°C, whereas for CQ8 and CQ103 strains, no differences were obtained at pH 5.5 and 6.0. In general, low pH, a_w, and temperature values increased the latency times and decreased the growth rate and colony diameter, although a_w and temperature were the most limiting factors. Maximum aflatoxin production on the cheese-based medium occurred at pH 5.0, 0.95 a_w, and 25 or 30°C, depending on the strain. This study shows the effect of pH, a_w, and temperature factors on growth and aflatoxin production of 3 aflatoxigenic A. flavus strains on a cheese-based medium. The findings may help to design control strategies during the cheesemaking process and storage, to prevent and avoid aflatoxin contamination by aflatoxigenic molds.     

Influence of substrate and low temperature on growth and survival of verotoxigenicEscherichia coli

Growth and survival of verotoxigenicEscherichia coli(VTEC) were studied in tryptic soy broth (TSB), brain–heart infusion (BHI), and whole milk at 12, 9.5, 8.5, 7.5, 6.5, and 5.5°C (±0.2°C). Colony forming units (cfu) were enumerated by surface plating on eosin methylene blue (EMB) agar plates and incubating at 37°C for 24 h. Growth curves were plotted and lag and generation times were determined. Results indicated that the average generation times for each strain was similar in each medium (4.6±0.9 h) at 12°C. However, at 9.5°C the average generation time differed significantly among the media (P<0.05). The average generation time was longer in BHI compared to TSB and whole milk at 9.5°C (14 h in BHI vs 11 h in milk and TSB). Also, the generation time of individual strains in BHI varied from 9.5–22 h at 9.5°C. Decrease of temperature to 8.5°C resulted in lack of growth of two strains in BHI. At 7.5°C none of the strains grew in BHI, one grew in TSB, whereas all grew in milk. A further decrease of temperature to 6.5°C did not allow growth of any of the strains in TSB or BHI whereas all grew in milk. Addition of 5% lactose to TSB and BHI enabled survival and growth of all strains at 6.5°C. The results of this study demonstrate the influence of growth medium composition on the minimum growth temperature of verotoxigenicE. coli.     

Effect of vanillin concentration,pH and incubation temperature onAspergillus flavus,Aspergillus niger,Aspergillus ochraceusandAspergillus parasiticusgrowth

The effects of incubation temperature (10–30°C), pH (3.0–4.0) and vanillin concentration (350–1200ppm) on the growth ofAspergillus flavus, Aspergillus niger, Aspergillus ochraceusandAspergillus parasiticuswere evaluated using potato–dextrose agar adjusted to water activity (a_w) 0.98. The radial growth rates after a lag period followed zero-order kinetics with constants that varied from 0 (no growth) to 0.63mmh⁻¹. The lag period depended on vanillin concentration, pH and incubation temperature. The germination time and the radial growth rates were significantly affected by the three studied variables (P<0.001). The inhibitory conditions (no growth after 30 days) depend on the type of mold. A niger, the most resistant species, was inhibited at 15°C, pH 3.0 and 1000ppm. ForA. ochraceus, the most sensitive, the inhibitory conditions in presence of 500ppm vanillin were pH 3.0 and temperature ≤25°C or pH 4.0 with temperature ≤15°C.     

Increased Aflatoxin Production by Aspergillus parasiticus Under Conditions of Cycling Temperatures

The effects of cycling temperatures (5°C for 12 hr and 25°C for 12 hr) on aflatoxin production by Aspergillus parasiticus NRRL 2999 in yeast extract sucrose (YES) medium were studied. Cycling temperatures, after preincubation at 25°C for various times, resulted in more aflatoxin B₁, G₁, and total aflatoxin production than did constant incubation at either 25°C, which is generally considered to be the optimum for aflatoxin production, or 15°C, which is the same total thermal input as the 5-25°C temperature cycling. With increased preincubation time at 25°C, toxin production increased and the lag phase of growth was shortened or not evident. Cultures that were preincubated at 25°C for 1, 2, and 3 days prior to onset of temperature cycling showed the greatest increase in maximum aflatoxin production over the 25°C and 15°C constant temperatures. Cultures that were not preincubated at 25°C but subjected to constantly fluctuating temperatures produced maximum amounts of aflatoxin equivalent to cultures incubated at a constant 25°C. The maximum aflatoxin production at all temperatures studied occurred during the late log phase of growth and at pH minimums. Aflatoxins were found in higher concentrations in the broth than the mycelia under temperature cycling conditions, at 15°C, and at 25°C during the first 21 days of incubation, whereas greater amounts of toxin were retained in mycelium at 25°C in the later incubation period (28-42 days).     

Synergistic effect of Sodium Chloride,temperature and pH on growth of a cocktail of spoilage yeasts: a research note

The effect of sodium chloride (0·5–10% w/v), pH (2·6–6·3) and temperature (1–22°C) were studied on the growth of a cocktail of food spoilage yeasts. The length of the lag phase and the time taken to reach the level of 10⁶cfu ml⁻¹were calculated for each set of conditions. It was found that the lag phase constituted as little as 21% of the total time to reach 10⁶cfu ml⁻¹when the yeasts were grown in favourable conditions and as much as 62% of the total time when more extreme conditions were used. It was concluded that the lag phase was the most important factor affecting the spoilage potential of chilled foods with low pH and high salt values. The single most effective factor in reducing the growth rate of yeasts was temperature. The lag phase was 15, 38, 270, 630 and 875 h when the temperature was 15, 8, 4, 2 and 1°C respectively. At any single temperature, there appeared to be a synergistic effect of NaCl and pH and under the most extreme conditions tested (1°C; pH 5·8; 6% NaCl), the lag phase was over 1000 h. These data have implications for the spoilage potential of high salt, reduced pH foods stored at chill temperatures.     

Turbidimetric definition of growth limits in probiotic Lactobacillus strains from the perspective of an adaptation strategy

The application of an adaptation strategy for probiotics, which may improve their stress tolerance, requires the identification of the growth range for each parameter tested. In this study, 4 probiotics (Lactobacillus acidophilus, Lacticaseibacillus casei, Lacticaseibacillus rhamnosus, and Lactiplantibacillus plantarum) were grown under different pH, NaCl, and sucrose concentrations at 25°C, 30°C, and 37°C. Turbidimetric growth curves were carried out and lag phase duration, maximum growth rate, and amplitude (i.e., the difference between initial and stationary phase optical density) were estimated. Moreover, cell morphology was observed, and cell length measured. The growth response, as well as the morphological changes, were quite different within the 4 species. The L. acidophilus was the most sensitive strain, whereas L. plantarum was shown to better tolerate a wide range of stressful conditions. Frequently, morphological changes occurred when the growth curve was delayed. Based on the results, ranges of environmental parameters are proposed that can be considered suboptimal for each strain, and therefore could be tested. The quantitative evaluation of the growth kinetics as well as the morphological observation of the cells can constitute useful support to the choice of the parameters to be used in an adaptation strategy, notwithstanding the need to verify the effect on viability both in model systems and in foods.     

The effect of a competitive microflora,pH and temperature on the growth kinetics of Escherichia coli O157:H7

Growth curves were generated for Escherichia coli O157:H7 in brain–heart infusion broth incubated at 37 or 15°C in the presence of individual and combinations of competing microflora. Broths were inoculated withE. coli O157:H7 (log₁₀3·00 cfu ml⁻¹) and competitors (log₁₀4·00 cfu ml⁻¹) and the initial pH of the broth was either neutral (7·0) or adjusted to 5·8 and then sequentially reduced to 4·8 over 10 h to simulate fermentation conditions. Growth curves were also generated for the competitors in these cultures, including Pseudomonas fragi, Hafnia alvei, Pediococcus acidilactici (pepperoni starter culture) and Brochothrix thermosphacta . Gompertz equations were fitted to the data and growth kinetics including lag phase duration, exponential growth rates and maximum population densities (MPD) calculated. In pure culture, the growth parameters for E. coli O157:H7 in neutral pH broths were significantly different from those recorded in simulated fermentation broths (P<0·05). The presence of competitors in the broth also had a significant effect on the growth kinetics of the pathogen. H. alvei significantly inhibited the growth (lag phase, growth rate and MPD) of E. coli O157:H7 at 37°C, neutral pH and outgrew the pathogen under these conditions. In neutral pH cultures, two other competitors, B. thermosphacta and P. acidilactici also inhibited the lag phase of the pathogen but had no effect on the other growth parameters. In simulated fermentation broths, the growth rate of E. coli O157:H7 was consistently slower and the MPD lower in the presence of a competitive microflora than when grown individually. At 15°C, only one competitor, P. fragi significantly inhibited the lag phase of the pathogen. The implications of these findings for food safety are discussed.     

Temperature modulates the production and activity of a metalloprotease from Pseudomonas fluorescens 07A in milk

This work evaluated the expression and activity of a metalloprotease released by Pseudomonas fluorescens 07A in milk. Low relative expression of the protease by the strain was observed after incubation for 12 h at 25°C while the strain was in the logarithmic growth phase. After 24 h, protease production significantly increased and remained constant for up to 48 h, a time range during which the strain remained in the stationary phase. Conversely, at refrigeration temperatures, at 12 h the strain was still in the lag phase and expressed the protease at higher levels than when the logarithmic phase was reached. Casein fractions were highly degraded by P. fluorescens 07A, the purified protease, and the bacterial pellet on d 7 of incubation at 25°C and to a lesser extent at 10°C for the sample incubated with the bacterium. Heat treatment at 90°C for 5 min completely inactivated the proteolytic activity of the purified protease and the bacterial pellet. This work contributes to the knowledge about the conditions of milk storage that influence the production and activity of this extracellular metalloprotease. The results demonstrate the need to find alternative strategies to control the synthesis and activity of proteolytic enzymes in the dairy industry to ensure the quality of processed products.     

THE INFLUENCE OF TEMPERATURE ON SOME PROPERTIES OF YEAST

The effects of temperature on the growth of yeast and on its metabolic activity in distiller's malt wort have been studied. In un-aerated fermentations, maximum yeast production takes place at about 30° C. whereas the growth rate in aerated cultures is highest at 35° C. The lag phase of the yeast studied fell from 6 hr. at 20° C. to 2·8 hr. at 25° C. and was not thereafter greatly affected by increases of temperature until 42° C. was reached, at which point growth ceased. Maltase activity was maximal at 25° C. when considered in terms of unit quantities of either yeast or fermenting wort, but the optimum temperature for initial fermentation velocity varied according to the time over which the measurement was made, being maximal at 40° C. for 0·5 hr., and at 35° C. for 2 hr. Alcohol production was highest at 25° C. whereas glycerol and higher alcohol formation took place optimally at 30° C.     

Temperature-dependent development and reproduction of the cowpea weevil,Callosobruchus maculatus F., in mungbean: Estimating a target temperature for its control using aeration cooling

The bruchid, Callosobruchus maculatus F., commonly known as the cowpea weevil, infests stored mungbean and other legumes. Aeration cooling has potential as a non-chemical means of managing this species in stored legumes. Population growth of C. maculatus in mungbean was investigated at nine constant temperatures (15, 17.5, 20, 22.5, 25, 27.5, 30, 32.5 and 35 °C) at 60% RH so that a target temperature for cooling could be estimated. We used two laboratory strains: Strain 1 and Strain 2 that had been in culture for 16–17 years and 1–2 years respectively. The results for the two strains were very similar. Egg to adult development occurred between 20 and 35 °C for Strain 1 and 17.5 and 35 °C for Strain 2. The optimal temperature for population growth was estimated to be 32.2 and 33.7 °C for Strains 1 and 2, respectively. The estimated lower threshold for population growth, i.e. the temperature at which population growth is zero, was 17.5 °C for Strain 1 compared with 17.1 °C for Strain 2. Based on our results, we recommend a target temperature of 17 °C for aeration cooling to manage C. maculatus infestations in mungbean during storage.     

Use of Response Surface Methodology to Study the Combined Effect of Salt, pH, and Temperature on the Growth of Food-Spoiling Halotolerant Yeast, Debaryomyces nepalensis NCYC 3413

This paper reports the interaction of salt (NaCl and KCl), initial pH, and temperature and their effects on the specific growth rate and lag phase of food spoiling halotolerant yeast, Debaryomyces nepalensis. The optimization of salt, initial pH, and temperature was carried out using response surface methodology based on central composite design. The mathematical model showed that salt has a significant effect on specific growth rate and lag phase of D. nepalensis. The optimal conditions of salt concentration, pH, and temperature of growth were found to be 0.3 M NaCl, 7.1, 26 °C and 0.6 M KCl, 5.6, 25°C, respectively. Under these conditions, a maximum specific growth rate of 0.41 and 0.5 h⁻¹ was observed in medium containing NaCl and KCl, respectively. Lag phase can be increased most effectively either by increasing salt concentrations or both by decreasing (≤20°C) or increasing the temperature (≥40°C) with moderate (1.5 M) or low salt concentration (0.5 M), respectively. Results show that D. nepalensis need to generate weak acids to maintain the intracellular pH under pH and saline stress conditions. The results obtained in this study will be helpful in using optimal conditions for the maximum growth of the strain for the production of certain metabolites like organic acids and glycerol and designing food preservation procedures.     

Synthesis of extracellular lipase by a strain ofPseudomonas fluorescensisolated from raw camel milk

Ten strains of psychrotrophic bacteria were isolated from raw camel milk and were arranged according to their lipase production. Lipolytic activities ranged between 0.26 to 3.43 meq of palmetic acid 100 g^-;1of bovine milk fat h^-;1.Pseudomonas fluorescensRM₄was the most active strain. This bacterium could grow and secrete lipase over a wide range of temperatures. The optimum temperature for growth was 37°C, and the maximum lipase production was at 25°C. Growth was maximal after 96 h of incubation at 25°C, and lipase activity was maximal at 72 h post-inoculation at 25°C (during the late logarithmic phase). Shaking of the cultures (100 rpm) led to an increase in both growth and enzyme activities. Pseudomonas fluorescensRM₄was able to grow and secrete lipase over a pH range of 5.5-;8.50. Synthesis of the enzyme appeared to be inducible because no enzyme was detected in the absence of organic nitrogen. Supplementation of the basal medium with milk proteins enhanced lipase production. Tryptophan and lysine induced enzyme synthesis most effectively. Addition of 4 g l^-;1of glucose to broth stimulated both growth and production of lipase. Beyond this level of supplementation, lipase activity was considerably depressed.     

Predictive model for growth of Clostridium perfringens during cooling of cooked cured chicken

Estimates of the growth kinetics of Clostridium perfringens from spores at temperatures applicable to the cooling of cooked cured chicken products are presented. A model for predicting relative growth of C. perfringens from spores during cooling of cured chicken is derived using a nonlinear mixed effects analysis of the data. This statistical procedure has not been used in the predictive microbiology literature that has been written for microbiologists. However, recently software systems have been including this statistical procedure. The primary growth curves, based on the stages of cell development, identify two parameters: (1) germination, outgrowth, and lag (GOL) time, or lag phase time; and (2) exponential growth rate, egr. The mixed effects model does not consider GOL and egr as constants, but as random variables that would, in all likelihood, differ for different cooling events with the same temperature. As such, it is estimated that the egr, for a given temperature, has a CV of approximately 19%. The model obtained by the mixed effects model is compared to the one obtained by the more traditional two-stage approach. The estimated parameters from the derived models are virtually the same. The model predicts, for example, a geometric mean relative growth of about 9·4 with an upper 95% confidence limit of 21·3 when cooling the product from 51°C to 12°C in 8 h, assuming log linear decline in temperature with time. C. perfringens growth from spores was not observed at a temperature of 12°C for up to 3 weeks.     

Quality Changes and Shelf-Life Prediction of a Fresh Fruit and Vegetable Purple Smoothie

The sensory, microbial, and bioactive quality changes of untreated (CTRL) and mild heat-treated (HT; 90 °C/45 s) smoothies were studied and modelled throughout storage (5, 15 and 25 °C). The overall acceptability was better preserved in HT samples being highly correlated (hierarchical clustering) with the flavour. The sensory quality data estimated smoothie shelf-life (CTRL/HT) of 18/55 (at 5 °C), 4.5/12 (at 15 °C) and 2.4/5.8 (at 25 °C) days. The yeast and mould growth rate was lower in HT compared to CTRL while a lag phase for mesophiles/psychrophiles was observed in HT-5/15 °C. HT and 5 °C storage stabilised the phenolic content. Ferric reducing antioxidant power reported the best correlation (R ² = 0.94) with the studied bioactive compounds, followed by ABTS (R ² = 0.81) while DPPH was the total antioxidant capacity method with the lowest adjustment (R ² = 0.49). Conclusively, modelling was used to estimate the shelf-life of a smoothie based on quality retention after a short-time, high-temperature heat treatment that better preserved microbial and nutritional quality during storage.     

VARIATION OF LAG TIME AND SPECIFIC GROWTH RATE AMONG 11 STRAINS OF SALMONELLA INOCULATED ONTO STERILE GROUND CHICKEN BREAST BURGERS AND INCUBATED AT 25C1

One strain of 11 serotypes or 11 strains of Salmonella, which were isolated from the ceca of broilers, were surveyed for their growth kinetics on sterile ground chicken breast burgers incubated at 25C to determine the variation of lag time and specific growth rate. Growth curves, four per strain, were fit to a two‐phase linear model to determine lag time (h) and specific growth rate (log₁₀/h). Repeatability of growth kinetics measurements for individual strains had a mean coefficient of variation of 11.7% for lag time (range: 5.8 to 17.3%) and a mean coefficient of variation of 6.7% for specific growth rate (range: 2.7 to 13.3%). Lag time among strains ranged from 2.2 to 3.1 h with a mean of 2.8 h for all strains, whereas specific growth rate among strains ranged from 0.3 to 0.38 log₁₀ per h with a mean of 0.35 log₁₀per h for all strains. One‐way analysis of variance indicated that lag time (P =0.029) and specific growth rate (P =0.025) differed slightly among strains. S. Haardt had a shorter (P < 0.05) lag time than S. Agona and S. Brandenburg, whereas the specific growth rate of S. Enteritidis was less than (P < 0.05) the specific growth rates of S. Typhimurium and S. Brandenburg. All other strains had similar lag times and specific growth rates. The coefficient of variation among strains was 9.4% for lag time and 5.7% for specific growth rate. These results indicate that there were only minor differences in the lag times and specific growth rates among the strains of Salmonella surveyed. Thus, the growth kinetic values obtained with one strain of Salmonella may be useful for predicting the growth of other strains of Salmonella for which data do not currently exist.     

Mathematical models to predict kinetic behavior and aflatoxin production of Aspergillus flavus under various temperature and water activity conditions

Mathematical models were developed to predict fungal growth and aflatoxin production of Aspergillus flavus. Fungal growth and aflatoxin concentrations were measured. The Baranyi model was fitted to fungal growth and toxin production data to calculate kinetic parameters. Quadratic polynomial and Gaussian models were then fitted to μ_max and LPD (lag phase duration) values. The ranges of temperature and a _w values showing a μ_max value increase were 15–35°C and 0.891–0.984, respectively. LPD was only observed when the temperature was 20–35°C with a _w=0.891?0.972. The μ_{max growth} value increased up to 35°C with \(b_w = 0.2\left( {b_w = \sqrt {1 - a_w } } \right)\) , then values declined. LPD_growth values increased as the b _w value increased. The μ_max value for aflatoxins increased up to 25°C, but decreased after 30°C, indicating that the developed models are useful for describing the kinetic behavior of Aspergillus flavus growth and aflatoxin production.