Temperature,water activity and gas composition effects on the growth and aflatoxin production by Aspergillus flavus on paddy

The main aim of this study was to evaluate the combined effects of temperature with water activity (a_w) and CO₂ with a_w on the growth and aflatoxin production by Aspergillus flavus Link on paddy. The effects of temperature (20–30 °C) and a_w (0.92–0.98) on the relationship between colony diameter and aflatoxin production, and the influence of a_w (0.92–0.98) and CO₂ (20–80%) on the growth and toxin production were studied using full factorial design. Colony diameters were regularly measured and aflatoxins were periodically analyzed using HPLC with fluorescence detector. The growth and aflatoxin formation increased with a_w at the temperatures studied, and toxin production was positively correlated with the incubation time and colony diameter. Except at 0.92 a_w, as much as 80% CO₂ failed to inhibit the growth of fungi completely. However, at all a_w levels studied the growth parameters as estimated by Baranyi function and aflatoxin were affected by the increment in CO₂ where growth rates and aflatoxin were negatively correlated with CO₂ while the lag phase durations were positively correlated with CO₂. Under 0.98 a_w, the atmosphere enriched with 20% and 80% CO₂ lead to at least 59% and 88% reduction in growth and 47% and 97% in the toxin production, respectively. At 0.95 a_w, the lag phases of both isolates in average increased by a factor of 1.7–12 when the CO₂ levels in the headspace were between 20 and 80% compared to the control. The growth rate and lag phase durations under the modified atmospheres were successfully described using a polynomial equation (R² > 0.97). The results of the study could form a basis of indicative guidelines on the possible control of A. flavus and aflatoxin in paddy during temporary storage prior to drying.     

Environmental factors affect efficacy of some essential oils and resveratrol to control growth and ochratoxin A production by Penicillium verrucosum and Aspergillus westerdijkiae on wheat grain

This study determined the efficacy of three essential oils (bay, clove and cinnamon oil) and the antioxidant resveratrol (0–500 μg g⁻¹) on the control of growth and ochratoxin A (OTA) production by Penicillium verrucosum and Aspergillus westerdijkiae (=A. ochraceus) under different water activity (a_w, 0.90, 0.95, 0.995), and temperature (15, 25 °C) conditions on irradiated wheat grain. The most effective treatment (resveratrol) was then tested on natural grain. The ED₅₀ values for growth inhibition by the oils were 200–300 μg g⁻¹ at the a_w and the temperatures tested. For resveratrol, this varied from <50 μg g⁻¹ at 0.90–0.95 a_w to >350 at 0.995a_w at both temperatures. The ED₅₀ values for the control of OTA were slightly lower than for control of growth, with approx. 200 μg g⁻¹ required for the oils and 50–100 μg g⁻¹ of the antioxidant, at 0.90/0.95a_w and both temperatures. In wet grain (0.995a_w), higher concentrations were required. For growth there were statistically significant effects of single-, two- and three-way interactions between treatments except for concentration×temperature and concentration×temperature×essential oil/antioxidant treatment. For OTA control, statistically significant treatments were a_w, temperature×a_w, concentration×temperature, treatment×concentration, and three-way interaction of concentration×a_w×treatment for P. verrucosum and A. westerdijkiae. Subsequent studies were done with the best treatment (resveratrol, 200 μg g⁻¹) on natural wheat grain with either P. verrucosum or A. westerdijkiae at 0.85–0.995a_w and 15/25 °C over 28 days storage. This showed that the populations of the mycotoxigenic species and OTA contamination could be reduced by >60% by this treatment at the end of the storage period.     

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.     

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.     

‘Combined effect of water activity, pH and additives on growth of Staphylococcus aureus in model salami systems’

The combined effect of pH, water activity (a_w) and certain additives (sodium nitrite and nitrate, polyphosphates, wine and pepper) on growth and survival of two strains of S. aureus in model salami systems at 25°C was investigated. These additives are usually used in Argentinean salami formulation. The combinations of a_w 0·925-pH 5·00, or a_w 0·915 - pH 5·50 or a_w 0·900 - pH 6·00, together with the addition of the additives, resulted in synergistic growth inhibition of both strains. Sodium nitrite and nitrate played an important role in the inhibition, while pepper, polyphosphates and wine had no effect in controlling the growth of these organisms.     

Novel approach to oxygen control in modified atmosphere packaging of bakery products

PDA plates inoculated with mould spores were packaged in air and in a mixture of CO₂/N₂ (60 : 40) with headspace oxygen adjusted to range from <0·05% to 10% (v/v) and stored at 25°C. Those packaged in air showed mould growth after 1–1·5 d, while those in the gas mixture showed growth between 4–6 d after headspace oxygen reached a minimum concentration of 0·4%. Uninoculated plates packaged in air showed no change in headspace oxygen concentration over 30 d at 25°C, while those in the gas mixture with initial headspace oxygen of <0·05% showed a steady increase of oxygen, through packaging film permeability, at an average rate of 0·06%/d. If uninoculated plates were packaged in air together with a sachet of oxygen absorbant called Ageless, the headspace oxygen was quickly reduced to <0·05% within 9 h and remained unchanged thereafter.Crusty rolls with a_w of 0·95 and pH of 5·62 developed mould after 5–6 d when packaged in air, 9–11 d in 100% N₂ and 16–18 d in CO₂/N₂ (60 : 40). However, when Ageless was incorporated into all the packages the product remained mould free for >60 d. This study shows that oxygen absorbants offer the bakery industry a viable alternative to gas flushing.     

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.     

In-package atmospheric cold plasma inactivation of Salmonella in freeze-dried pet foods: Effect of inoculum population,water activity,and storage

Recently, Salmonella has been linked to numerous low water activity (a_w) food recalls. As an emerging technology, atmospheric cold plasma (ACP) has the potential to reduce Salmonella in low a_w foods. The objective of this study was to evaluate the effects of initial inoculum population of Salmonella (~ 10, 9 and 8 log CFU/cm²), treatment time (2.5, 5, 7.5 and 10 min), water activity (0.13, 0.34, 0.54), and post-treatment storage (0, 7 and 14 days at 21 °C) on the reduction of cocktails of S. Typhimurium and S. Senftenberg on freeze-dried pet food treats. The top surface of pet food was spot inoculated with cocktails of Salmonella with initial cell populations of 10, 9 and 8 log CFU/cm². Inoculated samples were first equilibrated to target a_w values, packaged with modified atmosphere (78% N₂ + 21% O₂ + 1% CO₂), and then in-package ACP treatments were performed. The initial inoculum population, treatment time, and post-treatment storage had significant (P < 0.05) effects on Salmonella inactivation efficacy of ACP. Regardless of the initial inoculum population and storage time, Salmonella equilibrated to 0.13 a_w pet food exhibited the greatest resistant to ACP treatment. ACP treatment for 10 min followed by 7 days storage reduced the Salmonella cells in 0.54 a_w pet food, to below the limit of detection (>4.5 log reduction), when the initial inoculum population was 8.2 log CFU/cm². Regardless of a_w of petfood, peroxide and thiobarbituric acid reactive substance values of 10 min ACP-treated pet foods were significantly increased to 3.44 meq/ kg of fat and 1.53 mg MDA/ kg pet food from 0.87 meq/ kg of fat and 0.50 mg MDA/ kg pet food, respectively. Post-ACP treatment storage of 10 min ACP-treated samples for 14 days further increased the lipid oxidation to 2.58 from 1.53 mg MDA/ kg of pet food, irrespective of the a_w of samples. This study showed that the Salmonella reduction efficacy of in-package ACP treatment in low a_w environment is influenced by various process and product factors.Industrial relevanceFreeze drying is not considered a kill step to inactivate microorganisms on foods. In the future, in-package ACP technology could be used as a potential method for post-packaging decontamination of freeze-dried pet foods in industrial settings after obtaining necessary regulatory approvals.     

Use of response surface methodology in shelf life extension studies of a bakery product

Effects of water activity, pH, storage temperature, concentration of potassium sorbate, CO₂ concentration in the package headspace and inoculum level on growth of Aspergillus niger, the major spoilage isolate of English style crumpets, was studied on an agar model system using an experimental design termed Response Surface Methodology. The initial screening design indicated that a_w, pH, levels of potassium sorbate, CO₂ and storage temperature were significant variables. When a central composite rotatable design was subsequently applied to four variables with pH held constant at pH 6.5 only a_w, storage temperature and CO₂ level were found to have a significant effect on mold growth. Using multiple regression analysis, a second order polynomial model was derived and used to predict the number of days to visible mold growth under various combinations of the remaining three variables. Generation of contour plots enabled selection of levels for each variable to give a product with a desired mold free shelf life. Typically, crumpets with an a_w of 0.96, packaged in 62% CO₂ had a mold free shelf life of 20 d when stored at 20°C. The observed shelf life results agreed well with predicted values on Potato Dextrose Agar and demonstrated RSM as a powerful and elegant research tool when several variables are to be evaluated simultaneously.     

Impact of environment and interactions of Fusarium verticillioides and Fusarium proliferatum with Aspergillus parasiticus on fumonisin B1 and aflatoxins on maize grain

Fusarium verticillioides and F proliferatum isolates were inoculated in mixed cultures with Aspergillus parasiticus on irradiated maize grain at two different inoculum concentrations (2 × 10⁵ and 2 × 10² conidia g^?1 dry maize). The treatments were 0.93–0.98 water activity (a_w) and 15 and 25 °C for 28 days. A complex relationship was found between a_w, temperature, inoculum concentration and the interactions which took place between fumonisin and aflatoxin producers. In general, A parasiticus reduced F verticillioides and F proliferatum populations (by 6–36%) but did not affect fumonisin B₁ production by these species. In contrast, while the Fusarium species were not able to decrease A parasiticus populations, they significantly reduced aflatoxin B₁ accumulation (by 30–93%). © 2001 Society of Chemical Industry     

Water availability and calcium propionate affect fungal population and aflatoxins production in broiler finisher feed during storage

The aim of this study was to investigate the effects of calcium propionate, water activity (a_w) and incubation time on the total fungal count and aflatoxins B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂) production in the broiler finisher feed. The feed was added with calcium propionate (5 g kg^–1), adjusted to 0.85, 0.90 and 0.95 a_w and stored for 28 days at 25°C, analysing for mould growth and aflatoxins production every 7 days. Analysis of variance indicated that all the factors (preservative, a_w and storage time) alone and in combination significantly (p < 0.001) affected the total fungal count and aflatoxins production in the feed. Minimum total fungal counts (1.99 × 10² CFU g^–1) were observed in calcium propionate feed at 0.85 a_w on day 1 and the highest (4.36 × 10⁹ CFUs g^–1) in control sample at 0.95 a_w on day 28 of storage. During the storage period, AFB₁ content in control samples increased from 11.35 to 73.44, from 11.58 to 81.81 and from 11.54 to 102.68 ng g^–1, whereas in preserved feed the content of B₁ increased from 11.47 to 37.83, from 11.54 to 49.07 and from 11.20 to 53.14 ng g^–1 at 0.85, 0.90 and 0.95 a_w, respectively. Similar patterns were noted for AFB₂, AFG₁ and AFG₂ contents. All the aflatoxins readily increased over storage time; however, the increase was much slower in preserved feed that contained a lower amount of available water. This study reveals that calcium propionate addition to poultry litter along with water activity amelioration is an effective tool for controlling mould incidence and aflatoxin production in poultry feed.     

Phytochemical stability in dried apple and green tea functional products as related to moisture properties

In this study, apple products made with and without added green tea extract were freeze-dried and stored for up to 45 days at 30 °C in low and intermediate moisture environments (water activity, a_w, 0.11, 0.22, 0.32, 0.57, and 0.75). Kinetic models were developed for the changes in color and decreases in contents of selected green tea and apple monomeric and polymeric flavanols and ascorbic acid. Moisture isotherms were developed for each product. At various moisture levels, the glass transition temperature (T_g) was measured by DSC and water mobility by ¹H NMR. Chemical changes were related to T_g, a_w, and water mobility in the products.Phytochemical degradation occurred more rapidly at higher moisture contents, except for caffeine which was stable. In the product containing apple with green tea, the content of monomeric flavan-3-ols decreased by 34% and 39% after 45 days of storage at a_w of 0.56 and 0.75, respectively. Phytochemical degradation correlated with increasing a_w, T_g, and water mobility. This study showed that, in general, storage at a_w 0.75 most affected phytochemical stability and color.     

Influence of water activity and temperature on growth and mycotoxin production by Alternaria alternata on irradiated soya beans

The aim of this study was to determine the effects of water activity (a_w) (0.99-0.90), temperature (15, 25 and 30 °C) and their interactions on growth and alternariol (AOH) and alternariol monomethyl ether (AME) production by Alternaria alternata on irradiated soya beans. Maximum growth rates were obtained at 0.980 a_w and 25 °C. Minimum a_w level for growth was dependent on temperature. Both strains were able to grow at the lowest a_w assayed (0.90). Maximum amount of AOH was produced at 0.98 a_w but at different temperatures, 15 and 25 °C, for the strains RC 21 and RC 39 respectively. Maximum AME production was obtained at 0.98 a_w and 30 °C for both strains. The concentration range of both toxins varied considerably depending on a_w and temperature interactions. The two metabolites were produced over the temperature range 15 to 30 °C and a_w range 0.99 to 0.96. The limiting a_w for detectable mycotoxin production is slightly greater than that for growth. Two-dimensional profiles of a_w × temperature were developed from these data to identify areas where conditions indicate a significant risk from AOH and AME accumulation on soya bean. Knowledge of AOH and AME production under marginal or sub-optimal temperature and a_w conditions for growth can be important since improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality. This could present a hazard if the grain is used for human consumption or animal feedstuff.     

Ecophysiology of Fusarium temperatum isolated from maize in Argentina

The effect of water activity (a_w = 0.95, 0.98 and 0.995), temperature (15, 25 and 30°C), incubation time (7, 14, 21 and 28 days), and their interactions on growth and moniliformin (MON), beauvericin (BEA), fusaproliferin (FUS) and fumonisin B₁ (FB₁) production by two strains of Fusarium temperatum isolated from Argentinean maize were determined in vitro on sterile layers of maize grains. The results showed that there was a wide range of conditions for growth and mycotoxins production by F. temperatum. Both strains were found to grow faster with increasing a_w and at 30°C. In relation to mycotoxin production, the two strains produced more FUS than the other mycotoxins regardless of a_w or temperature evaluated (maximum = 50 000 μg g^?1). For FUS, MON and BEA, the maximum levels were observed at 0.98 a_w and 30°C (50 000, 5000 and 2000 μg g^?1 respectively). The lowest levels for these three mycotoxins were detected at 15°C and 0.95 a_w (1700 and 100 μg g^?1 for FUS and MON respectively), and at 0.98 a_w (400 μg g^?1 for BEA). The maximum levels of FB₁ were produced at 15°C and 0.98 a_w (1000 μg g^?1). At all a_w and temperatures combinations evaluated there was an increase in toxin concentrations with time incubation. The maximum levels were detected at 21 days. Statistical analyses of a_w, temperature, incubation time, and the two- and three-way interactions between them showed significant effects on mycotoxins production by F. temperatum. For its versatility on growth and mycotoxin production, F. temperatum represents a toxicological risk for maize in the field and also during grain 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.     

Evolution of grain microbiota during hermetic storage of corn (Zea mays L.)

Respiration of biotic components of the grain ecosystem generates self-modified atmospheres (oxygen reduced and carbon dioxide enriched) during hermetic storage. The effect of temperature, moisture content and modified atmospheres on the evolution of maize microbiota is not entirely known. In this study, corn grain samples were conditioned to different moisture contents (14.3, 16.5 and 18.4%) and hermetically stored in glass jars at 15, 25 and 35 °C. Grain samples were collected at different stages of modified atmosphere evolution of each experiment: T0 (O₂: 21% initial concentration); T1 (O₂: 10%); T2 (O₂: 0%); T3 (CO₂ maximum concentration stabilized). Microbiota was quantified with Petri dish counts using selective growth media for different microbial groups. Additionally, ethanol, acetic acid and lactic acid were measured for monitoring anaerobic activity. Results indicated that there was a high correlation between water activity (a_w) and the time to reach anaerobiosis (R² of 0.85), the maximum CO₂ concentration (R² of 0.86), and the reduction of filamentous fungi and bacterial counts during hermetic storage (R² of 0.72 and 0.48, respectively). A differential behavior of the hermetic storage was observed according to a_w of the grain, and a general conceptual model is offered for its understanding. It was concluded that modified atmospheres reduced or inhibited microbial growth in stored corn, and that a_w was the most influential factor in the time to reach anaerobiosis, maximum CO₂ concentration, and the filamentous fungi and bacterial counts reduction during hermetic storage.     

Modeling Growth Rate and Assessing Aflatoxins Production by Aspergillus flavus as a Function of Water Activity and Temperature on Polished and Brown Rice

Abstract: The aim of this study was to model the radial growth rate and to assess aflatoxin production by Aspergillus flavus as a function of water activity (a_w 0.82 to 0.92) and temperature (12 to 42 °C) on polished and brown rice. The growth of the fungi, expressed as colony diameter (mm) was measured daily, and the aflatoxins were analyzed using HPLC with a fluorescence detector. The growth rates were estimated using the primary model of Baranyi, which describes the change in colony radius as a function of time. Total of 2 secondary models were used to describe the combined effects of a_w and temperature on the growth rates. The models were validated using independent experimental data. Linear Arrhenius–Davey model proved to be the best predictor of A. flavus growth rates on polished and brown rice followed by polynomial model. The estimated optimal growth temperature was around 30 °C. A. flavus growth and aflatoxins were not detected at 0.82 a_w on polished rice while growth and aflatoxins were detected at this a_w between 25 and 35 °C on brown rice. The highest amounts of toxins were formed at the highest a_w values (0.90 to 0.92) at a temperature of 20 °C after 21 d of incubation on both types of rice. Nevertheless, the consistencies of toxin production within a wider range of a_w values occurred between 25 to 30 °C. Brown rice seems to support A. flavus growth and aflatoxin production more than the polished rice. Practical Application: The developed models can be used to estimate to what extent the change in grain ecosystem conditions affect the storage stability and safety of grains without the need for running long‐standing storage study. By monitoring the intergranular relative humidity and temperature at different locations in the storage facility and inputting these data into the models, it is directly possible to assess either the conditions are conductive for the growth of A. flavus or aflatoxin production.     

Potential use of phenolic antioxidants on peanut to control growth and aflatoxin B1 accumulation by Aspergillus flavus and Aspergillus parasiticus

Food‐grade antioxidants: butylated hydroxyanisole (BHA), propyl paraben (PP) and butylated hydroxytoluene (BHT) (10 and 20 mmol g^?1) and all the mixtures of these chemicals were tested for inhibitory activity on the growth of and aflatoxin B₁ (AFB₁) accumulation by Aspergillus parasiticus and A. flavus on irradiated (7 kGy) peanut grains. Also, the influence of these treatments was evaluated in different water conditions (0.982, 0.955, 0.937a_w) at 11 and 35 days of incubation at 28 °C. Water activity (a_w) affected the fungal growth, no fungal development was observed at the highest stress water condition (0.937a_w). Butylated hydroxyanisole at 10 mmol g^?1 level and all the mixtures with PP and/or BHT were significantly effective (P = 0.05) in increasing lag phase and reducing growth rate and colony forming units per gram of peanut of both Aspergillus section Flavi strains and AFB₁ accumulation. The application of BHA at concentrations of 20 mmol g^?1 alone or with PP and/or BHT totally inhibited fungal growth at 11 and 35 days of incubation. The results suggest that the addition of these chemical mixtures on peanut grains at low levels has potential to impact synergically on the control of Aspergillus section Flavi. Copyright © 2007 Society of Chemical Industry     

Effect of modified atmosphere and temperature abuse on the growth from spores and cereulide production of Bacillus weihenstephanensis in a cooked chilled meat sausage

The effect of modified atmosphere packaging (MAP) on the germination and growth of toxin producing psychrotolerant Bacillus spp is not well described. A model agar system mimicking a cooked meat product was used in initial experiments. Incubation at refrigeration temperature of 8 °C for 5 weeks of 26 Bacillus weihenstephanensis including two emetic toxin (cereulide) producing strains showed that B. weihenstephanensis is sensitive to MAP containing CO₂. The sensitivity to 20% CO₂ was dependent on strain and oxygen level, being increased when oxygen was excluded from the MAP. Growth from spores was observed at the earliest within 2 weeks when 20% CO₂ was combined with 2% O₂ and in 3 weeks when combined with “0”% O₂ (the remaining atmosphere was made up from N₂). Results were validated in a cooked meat sausage model for two non-emetic and one emetic B. weihenstephanensis strain. The packaging film oxygen transfer rates (OTR) were 1.3 and 40 ml/m²/24 h and the atmospheres were 2% O₂/20% CO₂ and “0”% O₂/20% CO₂. Oxygen availability had a large impact on the growth from spores in the MAP meat sausage, only the most oxygen restricted condition (OTR of 1.3 ml/m²/24 h and “0”% O₂/20 % CO₂) inhibited growth of the three strains during 4 weeks storage at 8 °C. Cereulide production was undetectable during storage at 8 °C irrespective of choice of the MAP (quantified by liquid chromatography mass spectrometry/mass spectrometry). MAP storage at 8 °C for 1 and 3 weeks followed by opening of packages and temperature abuse for 1.5 h daily at 20 °C during 1 week resulted in increased cell counts and variable cereulide production in the meat sausage. A pre-history at 8 °C for 1 week in MAP with OTR of 1.3 or 40 ml/m²/24 h and 2% O₂ resulted in cereulide concentrations of 0.816 – 1.353 µg/g meat sausage, while a pre-history under the most oxygen restricted condition (OTR of 1.3 ml/m²/24 h, “0”% O₂/20 % CO₂) resulted in minimal cereulide production (0.004 µg/g meat sausage) at abuse condition. Extension of MAP storage at 8 °C for 3 weeks followed by abuse resulted in a substantially reduced cereulide production.Data demonstrates that MAP can be used to inhibit growth of a psychrotolerant toxin producing Bacillus spp. during chill storage at 8 °C, and substantially reduce the risk of emetic food poisoning at abuse condition. Results are of relevance for improving safety of ready to eat processed chilled foods of extended durability.     

Microbial inactivation after high-pressure processing at 600 MPa in commercial meat products over its shelf life

The behaviour of spoilage and pathogenic microorganisms was evaluated after high-pressure treatment (600 MPa 6 min, 31 °C) and during chilled storage at 4 °C for up to 120 days of commercial meat products. The objective was to determine if this pressure treatment is a valid process to reduce the safety risks associated with Salmonella and Listeria monocytogenes, and if it effectively avoids or delays the growth of spoilage microorganisms during the chilled storage time evaluated. The meat products covered by this study were cooked meat products (sliced cooked ham, pH 6.25, a_w 0.978), dry cured meat products (sliced dry cured ham, pH 5.81, a_w 0.890), and raw marinated meats (sliced marinated beef loin, pH 5.88, a_w 0.985). HPP at 600 MPa for 6 min was an efficient method for avoiding the growth of yeasts and Enterobacteriaceae with a potential to produce off-flavours and for delaying the growth of lactic acid bacteria as spoilage microorganisms. HPP reduced the safety risks associated with Salmonella and L. monocytogenes in sliced marinated beef loin.