Modelling the effect of temperature and water activity in the growth boundaries of Aspergillus ochraceus and Aspergillus parasiticus

The aim of this work was to model the growth of Aspergillus parasiticus and Aspergillus ochraceus, both mycotoxin producers, near to the growth/no growth boundaries and validate those models in sterile maize grain, peanuts and coffee beans. Malt extract agar was adjusted to six different water activities: 0.93, 0.91, 0.89, 0.87, 0.85 and 0.80. Plates were incubated at 10, 15, 20, 25, 30, 37 and 42 °C. For each of the 42 conditions, 10 Petri dishes were inoculated. Both kinetic and probability models were applied to colony growth data. The results of the present study indicate that the developed probability modelling approach could be satisfactorily employed to quantify the combined effect of temperature and water activity on the growth responses of A. ochraceus and A. parasiticus. However, validation of kinetic results led to poor goodness of prediction. In this study, the validation samples were placed near to the expected boundaries of the models in order to test them under the worst situation. Probability of growth prediction under extreme growth conditions was somewhat compromised, but it can be considered acceptable.     

Effect of hot acidic fructose solution on caramelisation intermediates including colour,hydroxymethylfurfural and antioxidative activity changes

Fructose model solutions of various concentrations were adjusted to pH 3.0, then incubated at 75, 85, and 95 °C, respectively, for studying the reaction kinetics of colour development, hydroxymethylfurfural (HMF) production, and antioxidative activities change in the caramelisation reaction. Results showed that colour development, HMF production, DPPH radical scavenging activity and ABTS^·+ radical scavenging activity in the system increased linearly with a first-ordered kinetics on fructose concentration. Their values of Q₁₀ were 1.87, 4.48, 2.06, and 2.24, and activation energies were 66, 160, 77, and 86 kJ/mol, respectively.     

Effect of water activity, temperature and incubation time on growth and ochratoxin A production by Aspergillus niger and Aspergillus carbonarius on maize kernels

The aim of this study was to determine the effects of water activity (a_w) (0.92-0.98), temperature (5-45 °C) and incubation time (5-60 days) on growth and ochratoxin A (OTA) production by Aspergillus niger and Aspergillus carbonarius on maize kernels using a simple method. Colony diameters of both strains at 0.92 a_w were significantly lower than those at 0.96 and 0.98 a_w levels. The optimum growth temperature range for A. niger was 25-40 °C and for A. carbonarius 20-35 °C. A. niger produced OTA from 15 to 40 °C, and the highest OTA level was recorded at 15 °C. The concentration of OTA produced at 0.92 a_w was significantly lower than those at 0.96 and 0.98 a_w. A. carbonarius produced OTA from 15 to 35 °C and the maximum concentration was achieved at 15 °C, although not differing statistically from the concentration detected at 20 °C. At 0.98 a_w the OTA concentration was significantly higher than at 0.96 and 0.92 a_w. Our results show that maize supports both growth and OTA production by A. niger and A. carbonarius. The studied strains were able to produce OTA in maize kernels from the fifth day of incubation over a wide range of temperatures and water availabilities. Although the limit of quantification of our method was higher than that required for the analysis of OTA in food commodities, it has proved to be a useful and rapid way to detect OTA production by fungi inoculated onto natural substrates, in a similar way as for pure culture. Both species could be a source of OTA in this cereal in temperate and tropical zones of the world.     

Consumer acceptance of roasted peanuts affected by storage temperature and humidity conditions

Consumer acceptance and intensity ratings of roasted peanuts stored at temperatures of 23, 30, 35, and 40 °C, and water activities (a_w) of 0.33, 0.44, 0.54, 0.67 and 0.75 were determined over time. Consumer acceptance ratings, including overall, appearance, color, and texture were affected by storage water activity and time, but not by storage temperature. Consumer intensity ratings of crunchiness were affected by storage water activity and time, but not storage temperature. Aroma acceptance, flavor acceptance, and roasted peanutty and stale/oxidized/rancid intensity ratings of roasted peanuts were dependent on storage temperature, water activity and time.Shelf-life of roasted peanuts was predicted by all consumer attributes (R²>0.60) and was best predicted by aroma acceptance (R²=0.75). Using contour plots with ratings >5.0 for all acceptance attributes, the shelf-life of roasted peanuts stored at 23 °C and between 0.33 and 0.75 a_w was limited by overall acceptance and decreased by approximately 50% with each 0.1 a_w increase. At accelerated temperatures of 30, 35 and 40 °C, shelf-life of roasted peanuts was predominantly limited by flavor acceptance (>5.0), and to a lesser extent by overall acceptance (>5.0). Shelf-life of roasted peanuts stored at accelerated temperatures decreased by 50% or more with each 0.1 a_w increase.     

Modelling the effect of temperature and water activity on the growth of Aspergillus parasiticus on irradiated Argentinian flint maize

A full factorially designed experiment including storage temperature (10, 20, 30 and 37 °C) and water activity (0.88, 0.92 and 0.96) was undertaken to study the growth of Aspergillus parasiticus in maize samples. Kinetic parameters such as specific growth rate (μ), lag phase duration and maximum logarithmic increase were determined by fitting the Modified Gompertz equation to the viable mould count data (N in CFU/g) as a function of time collected in twelve experiments. The average coefficient of determination (R²) was 0.987, being the mean standard deviation of the estimate of 0.216 in units of log₁₀N. In the practical range of 10-30 °C, the relationship of the three kinetic parameters with temperature was described by second order polynomial expressions, whose parameters, in turn, depended on water activity. The combined or full model i.e., the Modified Gompertz model with its parameters expressed as a function of temperature and water activity, was able to predict log₁₀N with an average percentage error of 4.3, so agreement with the experimental data was highly satisfactory.In a simulation exercise, the full model was able to predict the viable mould count, given an initial value and grain temperature and water activity histories, with promising results for maize storage.     

Water activity and temperature effects on mycotoxin production by Alternaria alternata on a synthetic tomato medium

Alternaria spp. have been reported to be the most frequent fungal species invading tomatoes. Certain species, in particular the most common one, A. alternata, are capable of producing several mycotoxins in infected plants and in agricultural commodities. Alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TA) are some of the main Alternaria mycotoxins that can be found as contaminants of food. The objective of this study was to determine the effect of water activity (a_w, 0.904, 0.922, 0.954, and 0.982) and temperature (6, 15, 21 and 35 °C) on mycotoxin production on a synthetic tomato medium of a cocktail inoculum of five strains of A. alternata isolated from tomato fruits affected by Blackmould. The optimum AOH production occurred at 0.954 a_w after 28 days of incubation at 21 °C. A temperature of 21 °C was the most favourable for AOH synthesis at all a_w levels. The maximum concentration of AME was determined at 0.954 a_w and 35 °C. The optimum conditions for TA accumulation were 0.982 a_w and 21 °C. At the 0.904 a_w no growth or germination was registered at 6 °C and 15 °C over the whole incubation period. At 21 °C and 35 °C growth occurred slowly but none of the toxins were detected at this a_w level. In general, high a_w levels were favourable for mycotoxin production. None of the other toxins was detected at quantifiable levels at 6 °C after the whole incubation period. A storage temperature of 6 °C or below could be considered as safe for tomato fruits and high moisture tomato products (a_w > 0.95), in relation with Alternaria toxins. The results obtained here could be extrapolated to evaluate the risk of spoilage in tomato fruits and tomato products caused by this pathogen.     

Modelling the effect of temperature and water activity on the growth rate and growth/no growth interface of Byssochlamys fulva and Byssochlamys nivea

The purpose of the present study was to apply a modelling approach to define the growth rate and growth/no growth interface of Byssochlamys fulva and Byssochlamys nivea on a synthetic medium as a function of temperature and water activity. Both fungal species were grown on malt extract agar at different temperatures (10, 15, 20, 25, 30, 35, 40 and 45 °C) and a_w levels (0.88, 0.90, 0.92, 0.94, 0.96 and 0.99) for a period of 30 days. Growth responses were evaluated over time in terms of colony diameter changes. Growth data were fitted to the primary model of Baranyi and the resulting growth rates were further modeled as a function of temperature and water activity using the cardinal model with inflection (CMI) ( Rosso et al., 1993). A logistic regression quadratic polynomial model was also employed to predict the probability of growth over storage time. Estimated parameters for minimum, maximum and optimum temperatures for growth were 9.1 °C, 46.4 °C and 32.1 °C for B. fulva and 10.5 °C, 43.2 °C and 32.1 °C for B. nivea. The respective values for a_w were 0.893, 0.993 and 0.985 for B. fulva and 0.892, 0.992 and 0.984 for B. nivea. No growth was observed at 0.88 a_w regardless of temperature for both species, whereas B. nivea ascospores could not grow at 10 and 45 °C irrespective of a_w. Regarding growth boundaries, the degree of agreement between predictions and observations was >98% concordant for both species. The erroneously predicted growth cases were 1.4–4.2% false positive and 2.1–3.5% false negative for B. nivea and B. fulva, respectively. The developed logistic model was validated with two literature data sets as well as with data from independent experiments carried out on fruit juices. Validation results showed that agreement with literature data for growth was 25 out of 36 (69.4%) cases, whereas validation on fruit juice data failed in only 6 cases (5 false positives and 1 false negative) out of 128 cases.     

Sporulation boundaries and spore formation kinetics of Bacillus spp. as a function of temperature, pH and a(w)

Sporulation niches in the food chain are considered as a source of hazard and are not clearly identified. Determining the sporulation environmental boundaries could contribute to identify potential sporulation niches. Spore formation was determined in a Sporulation Mineral Buffer. The effect of incubation temperature, pH and water activity on time to one spore per mL, maximum sporulation rate and final spore concentration was investigated for a Bacillus weihenstephanensis and a Bacillus licheniformis strain.     

Effect of water activity and temperature on growth of Alternaria alternata on a synthetic tomato medium

Alternaria alternata is a toxigenic fungus, predominantly responsible for Blackmould of ripe tomato fruits, a disease frequently causing substantial losses of tomatoes, especially those used for canning. The objective of this study was to determine the effect of water activity (a_w, 0.904, 0.922, 0.954, 0.982) and temperature (6, 15, 21 and 35 °C) on germination and radial growth rate on a synthetic tomato medium of a cocktail inoculum of five strains of A. alternata isolated from tomato fruits affected by Blackmould. The shortest germination time (1.5 days) was observed at 0.982 a_w, both at 21 °C and 35 °C. The germination time increased with a reduction on a_w. The fastest growth rate was registered at 0.982 a_w and 21 °C (8.31 mm/day). Growth rates were higher when a_w increased. No growth or germination was observed at the lowest a_w level evaluated (0.904) after 100 days of incubation at 6 °C and 15 °C. A temperature of 6 °C caused a significant reduction in growth rates, even at the optimum a_w level. The knowledge on the ecophysiology of the fungus in this substrate is necessary to elaborate future strategies to prevent its development and evaluate the consumer health risk.     

Survival of Enterobacter sakazakii in infant cereal as affected by composition, water activity, and temperature

Enterobacter sakazakii infections in preterm neonates and infants have been epidemiologically associated with consumption of reconstituted powdered infant formula. The bacterium has been isolated from grain, infant cereals, and cereal factory environments. A study was done to determine the survival characteristics of E. sakazakii initially at populations of 0.31 and 5.03 logCFU/g of infant rice cereal (a(w) 0.30, 0.45-0.46, and 0.68-0.69). Cereal was stored at 4, 21, and 30 degrees C and populations were monitored for up to 12 months. Survival of the pathogen in infant rice, barley, oatmeal, and mixed grain cereals (a(w) 0.63-0.66, 0.76, or 0.82-0.83) initially containing a population of 4.93-5.64 logCFU/g and held at 4, 21, and 30 degrees C up to 24 weeks was determined. Populations decreased significantly (p < or = 0.05) in all cereals stored at 21 and 30 degrees C regardless of a(w). Increases in a(w) or storage temperature accelerated the rate of death of E. sakazakii in dry infant cereals. However, at an initial population of 0.31 logCFU/g, E. sakazakii survived in rice cereal (a(w) 0.30-0.69) for up to 12 months at all storage temperatures. Survival of E. sakazakii was not affected by the composition of dry infant rice, barley, mixed grain, and oatmeal cereals (initial a(w) 0.63-0.83) stored for up to 24 weeks at 4, 21, or 30 degrees C. This study demonstrated that E. sakazakii can survive for up to 12 months in infant cereals having a wide range of a(w) when storage is at temperatures simulating those to which they may be exposed during distribution, at retail, and in the home.     

Effect of water content and temperature on glucosinolate degradation kinetics in broccoli (Brassica oleracea var. italica)

The effect of water content and temperature on glucosinolate thermal degradation in broccoli (Brassica oleracea var. italica) was investigated. Broccoli was freeze dried obtaining batches with water content between 13% and 82% (a_w from 0.32 to 0.99). These samples were heated at different temperatures (from 60 to 120 °C) and glucosinolate levels were monitored. To rule out enzymatic breakdown, myrosinase was inactivated prior to heating. Degradation could be described by first-order kinetics for all glucosinolates and all water contents. In the temperature range 60–100 °C the sample with 13% showed the lowest degradation rate, whereas at 120 °C the degradation rate increased with the water content. This particular behavior was reflected by the high activation energy value of the driest sample. Several hypotheses to explain the observed behavior are discussed.     

Effects of temperature and pH on the kinetics of caramelisation,protein cross-linking and Maillard reactions in aqueous model systems

Solutions of either glucose, bovine serum albumin (BSA), casein, glucose–BSA or glucose–casein were heated at 60 to 100 °C at pH 8.0 and pH 9.7, and the kinetics of the reaction markers (disappearance of the glucose and amino groups, development of UV absorbance and browning, and protein reticulation) were monitored. All the markers were enhanced at increasing temperatures, and, except for the disappearance of the free amino group and protein polymerisation, at alkaline pH levels. The loss of the amino group occurred in parallel with increase in polymerisation. The two proteins reacted in line with their amino content and solubility. The activation energy (E_a) of the amino group loss in the protein–glucose mixture was around 100 kJ per mol and that in the protein heated alone was higher. The E_a associated with the disappearance of glucose was around 90 kJ per mol. The E_a of UV absorbance and browning processes, which showed parallel time courses, were found to range from 90 to 150 kJ per mol.     

Effects of storage temperature and water activity on the survival of bifidobacteria in powder form

Effects of water activity and storage temperature on survival of bifidobacteria in powder form were investigated and kinetic analyses were performed to reveal characteristics of the stability. A significant positive correlation was observed between water activity and natural logarithm of the inactivation rate constant of bifidobacteria powder, indicating that higher water activity induced lower stability of bifidobacteria in powder form. Also, higher temperature condition induced lower survival rate, which was supported by that the stability was followed the Arrhenius theory. These findings constructed a prediction model for bifidobacteria survival in powder form.     

Modeling the influence of water activity and ascospore age on the growth of Neosartorya fischeri in pineapple juice

The ascospores of resistant fungi, Neosartorya fischeri, can survive commercial pasteurization, diminishing the shelf life of these products. The time that the ascospores remain in the environment and the effect that they can cause on mold growth are still unknown. This study is aimed to evaluate the influence of water activity (a_w) from 0.90 to 0.99 and the ascospore age (I) from 30 to 90 days of vitro incubation on the growth of N. fischeri in pineapple juice by mathematical modeling. The growth parameters on pineapple juice: adaptation phase (λ), maximum specific growth rate (μ_max) and maximum diameter reached by the colony (A) were obtained by fitting Modified Gompertz and Logistic models to the experimental data. Both models were able to describe microbial growth in pineapple juice, but the Modified Gompertz model presented a slightly superior performance based on statistical indices (correlation coefficients (R²), mean square error (MSE), Bias Factor and Accuracy Factor). The minimum values of λ and A, calculated by the Modified Gompertz model, were 64.7 h and 6.3 mm, while the maximum values were 178.2 h and 20.8 mm, respectively. The result showed that ascospore age did not influence the growth but a_w was statistically significant to the growth parameters λ and A.     

Comparisons of water activity and temperature impacts on growth of Fusarium langsethiae strains from northern Europe on oat-based media

This study has examined the effect of water activity (a_w, 0.995-0.90) and temperature (10-37 °C) on the lag phases prior to growth, growth rates and used models to develop two dimensional profiles for optimum and marginal conditions for two strains of Fusarium langsethiae from four northern European countries (UK, Norway, Sweden, and Finland) on an oat-based medium. Results showed that the optimum a_w for growth was at 0.98-0.995 and 25 °C. The limit for growth of the strains was at 0.92-0.93 a_w with minima of 10 °C. No growth occurred at 37 °C. The lag phases prior to growth were lowest under optimum conditions and extended to > 10 days at marginal conditions. Statistical analyses of intra and inter-strain differences in terms of both lag phases prior to growth and growth rates were not statistically significant. However, a_w and temperature were statistically significant factors. Two dimensional profiles for strains from each country of origin were built to identify optimum and marginal conditions for F. langsethiae for the first time. These environmental profiles will be beneficial for improving the ecological knowledge of this species which is able to produce trichothecene mycotoxins in a range of temperate cereals.     

The mycobiota of three dry-cured meat products from Slovenia

The surface mycobiota of three types of Slovenian dry-cured meat products were isolated from a total of 75 items of product that were sampled periodically during the drying/ripening stage of processing. The predominant filamentous fungal genus isolated was Penicillium. Eurotium spp., Aspergillus versicolor and Cladosporium spp. were isolated from only two of the products. Eight Penicillium species were identified. Penicillium nordicum was recovered frequently. Penicillium nalgiovense was recovered less frequently, from one product only (a salami), while a yet-to-be described species Penicillium “milanense” was isolated from 21 items. The other penicillia were rarely isolated. Of the isolated and identified species, those that can produce mycotoxins are: A. versicolor, Penicillium brevicompactum, Penicillium chrysogenum, P. nordicum, and Penicillium polonicum. Their growth on dry-cured meat products is undesirable.     

Improvement of the oral bioavailability of coenzyme Q10 by emulsification with fats and emulsifiers used in the food industry

Coenzyme Q₁₀ (CoQ₁₀) was emulsified with fats and emulsifiers commonly used in the food industry, and the oral bioavailability of the emulsified product was compared with that of a standard commercial product. Five commonly used fats (olive oil, safflower oil, coconut oil, butter, and cocoa butter), four types of emulsifier (lecithin, monoglycerides, calcium stearoyl-2-lactate (CSL), and diacetyl tartaric acid esters of monoglycerides), and two types of aqueous phase (distilled water with or without 8 g/100 g w/w skim milk) were employed in this study. It was found that CoQ₁₀ was more soluble in coconut and safflower oils than in the other fats. In addition, it was demonstrated that emulsion with coconut oil, skim milk aqueous solution, and CSL produced an optimal formulation. Based on the results, a model emulsified CoQ₁₀ product was prepared. Its oral bioavailability was confirmed to be slightly greater than that of a standard commercial CoQ₁₀ product.     

The influence of non-lethal temperature on the rate of inactivation of vegetative bacteria in inimical environments may be independent of bacterial species

The influence of non-lethal temperature on the survival of two species of food-borne bacteria under growth-preventing pH and water activity conditions was investigated. Specifically, inactivation rates of four strains of Escherichia coli and three strains of Listeria monocytogenes were determined in culture broth adjusted to pH 3.5 and water activity 0.90, to prevent growth of both species, and for temperatures in the range 5–45 °C at 5 °C intervals. Sixty-three inactivation rates were obtained, plotted on Arrhenius co-ordinates, and lines of best-fit determined by simple linear regression. Differences in the mean inactivation rate of each species at a given temperature were not significant (p < 0.05) with the exception of the rates at 25 °C. The inactivation rate responses of both species were comparable to those reported by McQuestin et al. (Appl. Environ. Microbiol., 75:6963–6972, 2009) for a variety of E. coli strains under a wide range of growth-preventing pH and water activity conditions. The results support the hypothesis that non-lethal temperature is a key factor governing the rate of inactivation of vegetative bacteria in foods when other hurdles prevent their growth and indicate that the temperature effect may also be independent of bacterial species.     

Complex regulation of the aflatoxin biosynthesis gene cluster of Aspergillus flavus in relation to various combinations of water activity and temperature

A microarray analysis was performed to study the effect of varying combinations of water activity and temperature on the activation of aflatoxin biosynthesis genes in Aspergillus flavus grown on YES medium. Generally A. flavus showed expression of the aflatoxin biosynthetic genes at all parameter combinations tested. Certain combinations of a_w and temperature, especially combinations which imposed stress on the fungus resulted in a significant reduction of the growth rate. At these conditions induction of the whole aflatoxin biosynthesis gene cluster occurred, however the produced aflatoxin B₁ was low. At all other combinations (25 °C/0.95 and 0.99; 30 °C/0.95 and 0.99; 35 °C/0.95 and 0.99) a reduced basal level of cluster gene expression occurred. At these combinations a high growth rate was obtained as well as high aflatoxin production. When single genes were compared, two groups with different expression profiles in relation to water activity/temperature combinations occurred. These two groups were co-ordinately localized within the aflatoxin gene cluster. The ratio of aflR/aflJ expression was correlated with increased aflatoxin biosynthesis.