Modeling of germination and growth of ochratoxigenic isolates of Aspergillus ochraceus as affected by water activity and temperature on a barley-based medium

Aspergillus ochraceus is an ochratoxin producing fungus that can be found on stored cereal grains such as barley. The objective of this study was to determine the effects of water activity (a_w, 0.75–0.99) and temperature (10–30°C) on germination and growth on barley extract agar medium (BMEA) of three isolates of Aspergillus ochraceus. The three isolates showed an optimal a_w for germination and growth of 0.99–0.95 at 20–30°C, with a marked increase of the lag phases and decrease of germination and growth rates at the marginal levels of a_w and temperature assayed. Minimum level of a_w for germination was 0.80 and 0.85 for growth. Data were then modeled by an MLR regression and response surface models were obtained. These models may allow a rough prediction of germination/growth as a function of the storage temperature and moisture content of barley grains.     

Modelling of effects of water activity and temperature on germination and growth of ochratoxigenic isolates of Aspergillus ochraceus on a green coffee-based medium

Influence of water activity (0.75-0.99 a(w)) and temperature (10, 20 and 30 degrees C) on germination and mycelial growth on green coffee extract agar medium of three ochratoxigenic isolates of Aspergillus ochraceus was studied. Optimal conditions for germination and growth were observed at 0.95-0.99 a(w) and 20-30 degrees C for the three isolates. Minimum a(w) level for germination was 0.80, and 0.85 for mycelial growth. At marginal a(w) and temperature levels assayed, the lag phases prior to germination increased and the growth rates showed a significant decrease in comparison with the optimal conditions. Data were modelled by a multiple linear regression (MLR) and response surface models were obtained. Germination and growth of A. ochraceus in green coffee beans could be prevented or at least inhibited to some extent by minimising the time that coffee beans are exposed to temperature and humidity conditions near to the optimum during processing and storage. This could be an empirical approach to predict the effects of water activity and temperature conditions on the development of ochratoxigenic isolates of A. ochraceus during handling and storage of green coffee.     

Influence of water activity and temperature on conidial germination and mycelial growth of ochratoxigenic isolates of Aspergillus ochraceus on grape juice synthetic medium. Predictive models

The first stages in the development of Aspergillus ochraceus, an ochratoxin A‐producing fungus that infects grapes and may grow on them, have been studied on a synthetic nutrient medium similar to grape in composition. Spore germination and mycelial growth have been tested over a water activity (a_w) and temperature range which could approximate to the real conditions of fungal development on grapes. Optimal germination and growth were observed at 30 °C for all three isolates tested. Maximal germination rates were detected at 0.96–0.99 a_w at 20 °C, while at 10 and 30 °C the germination rates were significantly higher at 0.99 a_w. Although this abiotic factor (a_w) had no significant influence on mycelial growth, growth rates obtained at 0.98 a_w were slight higher than those at other a_w levels. Predictive models for the lag phase before spore germination as a function of water activity and temperature have been obtained by polynomial multiple linear regression, and the resulting response surface models have been plotted. Copyright © 2005 Society of Chemical Industry     

Ecophysiology of ochratoxigenic Aspergillus ochraceus and Penicillium verrucosum isolates. Predictive models for fungal spoilage prevention - a review

Ochratoxin A (OTA) is a secondary metabolite produced by several species of Aspergillus and Penicillium; among them Aspergillus ochraceus and Penicillium verrucosum are two ochratoxigenic species capable of growing in different climates and thus contamination of food crops with OTA can occur worldwide. OTA can be found in a wide range of foods such as cereals, coffee, cocoa, spices, beer, wine, dried vine fruit, grapes and meat products. OTA is toxic to animals, it presents neurotoxic, immunotoxic and nephrotoxic effects. It has been implicated in a human kidney disorder known as Balkan Endemic Nephropathy. This review focuses on the ecophysiology of ochratoxin-producing Aspergillus ochraceus and Penicillium verrucosum, the effect of environmental factors on their germination, mycelial growth, and OTA production. Knowledge of environmental conditions required for sucessive stages of fungal development represent the first step towards preventing mycotoxin formation. Predictive models for different stages of fungal development are presented, which allow prediction of the time before spoilage as a function of the abiotic factors. Finally, the implications of these studies in management of barley, coffee and grapes are described. This can help to identify the critical control points in their production, storage and distribution processes.     

Effect of water activity and temperature on growth of ochratoxigenic strains of Aspergillus carbonarius isolated from Argentinean dried vine fruits

Aspergillus carbonarius is an ochratoxin A (OTA) producing fungus, predominantly responsible for the production of this mycotoxin in grapes, wine and dried vine fruits. The objective of this study was to determine the in vitro effects of water activity (a(w), 0.80-0.95) and temperature (15-35 degrees C) on lag phase extension and radial growth rate of a cocktail inoculum of four strains of A. carbonarius. The maximum growth rate was observed at 0.95 a(w) and 30 degrees C (17.46 mm day(-1)). In general, growth rates increased with the increment of a(w). No growth was observed at a(w) below 0.85. For all a(w) levels tested the highest growth rate was detected at 30 degrees C. At 15 degrees C growth only occurred at the higher a(w) levels evaluated (0.925 and 0.95) at a growth rate of 3.82 and 5.57 mm day(-1) respectively. The shortest lag phase (0.26 days) was found at 0.925 a(w) and 35 degrees C. At marginal conditions of a(w) and temperature the lag phases increased, being the highest registered at 20 degrees C and 0.89 a(w) (33.7 days). The pattern of effects of environmental factors on growth was similar among Argentinean A. carbonarius strains and those from several European countries, Israel and Australia.     

Impact of relative humidity and temperature on visible fungal growth and OTA production of ochratoxigenic Aspergillus ochraceus isolates on grapes

Aspergillus ochraceus is an ochratoxin A (OTA) producer mould found in grapes and this may contribute to OTA contamination in wines and grape juices. The influence of relative humidity (R.H.; 80, 90 and 100%) and temperature (10, 20 and 30 °C) on visible mould growth on grapes and OTA accumulation after 14 days of incubation by this fungal species has been studied using a full factorial design with three replicates.The two abiotic factors and their interaction (R.H.×temperature) affected significantly the A. ochraceus growth in berries, which was maximum at 90–100% R.H. levels. With regard to the optimum temperature level, it occurred at 30 °C at 80 and 90% R.H., whereas no significant differences were detected at 20 and 30 °C when R.H. was 100%.OTA production by A. ochraceus on grapes was not significantly modified by the assayed levels of temperature and R.H, with a mean value of 3.53 ng g⁻¹.Predictive models of percentage of grapes with visible growth of A. ochraceus isolates under different relative humidity and temperature are presented.     

Effects of substrate, water activity, and temperature on growth and verrucosidin production by Penicillium polonicum isolated from dry-cured ham

Penicillium polonicum, a common mold on dry-cured meat products, is able to produce verrucosidin, a potent neurotoxin. The ability of P. polonicum isolated from dry-cured ham to grow and produce verrucosidin from 4 to 40 degrees C at water activities (a(w)) of 0.99, 0.97, and 0.95 on malt extract agar (MEA) and a medium made up with meat extract, peptone, and agar (MPA) was evaluated. Verrucosidin was quantified by high-pressure liquid chromatography and mass spectrometry. P. polonicum was able to grow on MEA and MPA at all the a(w) values tested from 4 to 37 degrees C but not at 40 degrees C. The optimal environmental conditions for growth were 20 degrees C, 0.99 a(w) on MEA and 20 to 25 degrees C, 0.97 a(w) on MPA, but the highest amount of verrucosidin was obtained at 25 degrees C, 0.99 a(w) in both media. No direct correlation between extension of mold growth and verrucosidin production was found. Temperature appears to be the most important factor ruling mycelial growth, whereas verrucosidin accumulation is mostly influenced by a(w). However, analysis of variance of the data showed that there was a complex interaction among all the environmental factors (medium, temperature, and a(w)) that significantly (P < 0.0001) affected growth and verrucosidin production. The reduction of a(w) to intermediates values of 0.95 has a stronger effect on growth on MEA than on MPA. Given that the meat-based medium proved to be an appropriate substrate for the biosynthesis of verrucosidin by P. polonicum, the ability of this mold to produce the toxin on meat products should be established.     

Distributions of the growth rate of the germ tubes and germination time of Penicillium chrysogenum conidia depend on water activity

The effects of water activities for sporulation (a(wsp)) and germination (a(wge)) on the distributions of the growth rate of the germ tubes (mu) and the germination time (t(G)) of Penicillium chrysogenum conidia were determined by monitoring the length of the same germ tubes throughout the experiments automatically. No relationship between the individual t(G)'s and mu's could be established. Irrespective of the water activity for germination, mu was greater and t(G) was less for conidia produced at 0.95a(wsp) than that at 0.99a(wsp). At 0.99 a(wge) the mean and the standard deviation of t(G) were smaller than those obtained at 0.95a(wge). At 0.99a(wge), normal distributions for mu and t(G) were exhibited, but not at 0.95a(wge). The cumulative frequencies were used to reconstruct the germination curves. Great differences in the percentage of spores capable of germination (P(G)) and in the mean germination times between conidia produced at 0.95a(wsp) and at 0.99a(wsp) were clearly exhibited at 0.95a(wge), thus demonstrating the paramount influence of sporulation conditions on germination kinetics.     

Environmental factors and weak organic acid interactions have differential effects on control of growth and ochratoxin A production by Penicillium verrucosum isolates in bread

Studies were conducted to examine the effect of water activity (a(w), 0.97, 0.95, 0.93), pH (4.5, 6.0) and preservative type and concentration (potassium sorbate (KS), calcium propionate (CaP), 300 and 3000 ppm) on colonisation and ochratoxin A (OTA) production by three strains of Penicillium verrucosum on bread analogues. At 25 degrees C, colony extension from a point source was similar for all three strains at the different environmental treatments examined, with optimum growth at 0.97 and 0.96 a(w) and pH 6. Temporal OTA production over 36 days varied significantly with strain, pH and a(w) level. Maximum OTA was produced at approximately 28-36 days for two of the strains with an optimum at 0.93 a(w) at pH 6.0. One strain produced 75-80 microg g(-1) analogue under these conditions. However, at pH 4.5, optimum OTA was produced at 0.95 a(w) after a similar time period on bread analogues, with maximum amounts of about 25-35 microg g(-1) bread. KS and CaP incorporation inhibited growth completely at pH 4.5 and 3000 ppm at all three a(w) levels. However, at suboptimal concentration, little growth inhibition occurred. At pH 6, even with 3000 ppm, growth of all three strains occurred. Where effective inhibition of growth was achieved (3000 ppm/pH 4.5, all a(w) levels), no OTA was detected. However, when 300 ppm was used, up to 7 microg g(-1) was detected in bread analogues at pH 4.5. At pH 6, OTA was produced at all treatment conditions by all three strains, although preservative-treated bread had less present. Statistical analysis showed that many of the one-, two- and three-way interactions had a significant effect on growth and OTA production. These results suggest that there is a significant risk from OTA contamination if lower concentrations of existing preservatives are used in wheat-based bakery products contaminated by spoilage mycotoxigenic moulds such as P. verrucosum.     

Effect of water activity and temperature on growth of three Penicillium species and Aspergillus flavus on a sponge cake analogue.

This study compared the effect of temperature and water activity and their interactions on the rate of mycelial growth of Penicillium aurantiogriseum, P. chrysogenum, P. corylophilum and Aspergillus flavus on a sponge cake analogue. As expected, growth rates showed dependence on a(w), and temperature. However, no significant differences were observed in the growth rates of different isolates. The minimum a(w) values for growth of the Penicillium spp. was 0.85-0.90. A. flavus was able to grow at 0.90 a(w) when the temperature was above 15 degrees C. This study has shown that fungal growth by these species on a sponge cake analogue, with a composition similar to usual bakery products, is prevented if the a(w) is kept at < 0.85.     

Effects of selected natural preservatives on the mycelial growth and ochratoxin A production of the food-related moulds Aspergillus westerdijkiae and Penicillium verrucosum

ABSTRACT

The present study examines the influence of the natural preservatives carvacrol, eugenol, trans-cinnamaldehyde and the essential oil (EO) Origanum vulgare on ochratoxin A (OTA) production and the mycelial growth of two food-related moulds, Penicillium verrucosum and Aspergillus westerdijkiae, by broth macro-dilution assay for 21 days. With the addition of ½ minimum inhibitory concentration (MIC) carvacrol, eugenol and O. vulgare EO, the mycelial dry weight of both moulds decreased significantly over the whole incubation period of 7, 14 and 21 days. Trans-cinnamaldehyde slightly stimulated the growth of A. westerdijkiae and P. verrucosum at 14 and 21 days of incubation. Growth inhibition did not accompany inhibition of OTA production. Although the growth of both moulds was inhibited after the addition of ½ MIC carvacrol, eugenol and O. vulgare EO, the OTA production of the strong mycotoxin producer A. westerdijkiae was stimulated. Only trans-cinnamaldehyde inhibited the production of OTA with the addition of ½ MIC. P. verrucosum produced significantly less OTA than A. westerdijkiae, and its mycotoxin production was almost completely inhibited by the addition of ½ MIC of the natural preservatives.     

萌发对青稞甜醅饮料的活性成分及其抗氧化活性的影响

目的 探究青稞籽粒萌发对所制青稞甜醅饮料的活性成分及其抗氧化活性的影响。方法 对青稞籽粒进行不同时间的萌发预处理, 再进行甜醅发酵并与纯净水、稳定剂复配制备青稞甜醅饮料, 以未萌发青稞籽粒制备的饮料为对照, 测定饮料的总酚、总黄酮含量及对2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐[2-azinlbis-(3-ethylbenzthiazoline-6-sulphonate), ABTS]、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)和羟自由基的清除率。结果 不同时间萌发时间处理条件下, 青稞甜醅饮料的活性成分含量及其抗氧化活性相较于对照均有所提升, 其中总酚和总黄酮含量均在萌发第40 h时达到最高, 分别为2.69 mg/mL和1.71 mg/mL, 分别比对照 2.21和1.35 mg/mL提高了21.72%和26.67%。经萌发处理后的青稞甜醅饮料对ABTS自由基和羟自由基清除能力在萌发第48 h时清除率分别达到64.64%和76.97%, 分别比对照48.83%和67.61%提高了32.38%和13.84%, 对DPPH自由基清除能力在萌发第40 h时清除率达到最高为94.11%, 比对照86.28%提高了9.07%。结论 青稞籽粒经萌发处理后制得的青稞甜醅饮料具有良好的抗氧化能力, 为提高青稞系列产品的附加值提供参考。     

Predicting fungal growth: the effect of water activity on Penicillium roqueforti.

The effect of water activity on the colony growth of Penicillium roqueforti is studied by predictive modelling techniques. Measured colony diameter growth curves are fitted to estimate the growth rate and lag phase of the curves. The colony growth rate was modelled by a quadratic function of transformed water activity (a(w)) values, as suggested by Baranyi et al. (Food Microbiol. 10 (1993) 43-59). The lag time was modelled as a function of water activity, by means of the sum of a constant term and a hyperboloid function of a(w) raised to the second power. The lag-phase of Penicillium roqueforti was found insensitive to the water activity in the range of its higher (a(w) > 0.92) values.     

桔青霉在小麦中最大比生长速率的温度-水分活度复合模型的建立

为预测引起小麦储藏霉菌桔青霉的生长,在15～35℃温度下,0.70～0.96水分活度(Aw)下进行小麦储藏模拟实验,建立桔青霉P21的温度-水分活度预测模型。结果表明,桔青霉P21最适生长条件为温度25℃,Aw 0.85。一级生长模型中Boltsmann模型和Logistic模型拟合度都达到0.99。以Logistic模型拟合数据建立二级模型,采用平方根模型和Arrhenius,Davey模型拟合,Arrhenius,Davey模型能较好描述桔青霉P21最大比生长速率与温度、水分活度之间的对应关系,相关系数R达0.9440。     

Prediction of fungal growth and ochratoxin A production by Aspergillus ochraceus on irradiated barley grain as influenced by temperature and water activity

Ochratoxin A (OTA) is a secondary metabolite of Aspergillus and Penicillium species, including Aspergillus ochraceus, a species that can be found in stored cereal grains such as barley. The objective of this study was to determine the effects of water activity (a(w), 0.80-0.99), temperature (10, 20, 30 degrees C), and A. ochraceus isolate differences on radial growth and OTA production in irradiated barley grains. The three isolates showed optimal conditions for growth and ochratoxin A production at 0.99 a(w) and 30 degrees C, with a marked decrease of growth rates and OTA production at the lowest levels of a(w) and temperature assayed. The minimum a(w) level for growth, observed in this study, was 0.85 and 0.90 a(w) for OTA production. Significant differences among the isolates were found. Lag phases prior to fungal growth and OTA production values were modelled by multiple linear regression and response surface models. These models could provide an approximate prediction of growth and OTA production.     

Mould growth on traditional greek sausages and penicillin production by Penicillium isolates

Visible moulds were isolated and identified from traditional Greek sausages from Northern Greece. Penicillium species were isolated from 90.8% of visibly mouldy sausages. Penicillium solitum, P. nalgiovense and P. commune species made up 60.6% of the total number of isolates. The most frequently occurring species was P. solitum (26.1%). P. nalgiovense and P. olsonii were found to be positive to penicillin production in an agar assay and further examination for antibiotic production in liquid culture with complex media designed for penicillin production, confirmed their ability for penicillin biosynthesis. Penicillin production by P. olsonii is reported for the first time in this study.     

Effect of water activity and temperature on mycelial growth and ochratoxin A production by isolates of Aspergillus ochraceus on irradiated green coffee beans

Aspergillus ochraceus as a fungal contaminant and ochratoxin A (OTA) producer plays an important role in coffee quality. Temperature and water activity (a(w)) significantly influence mycelial growth and OTA production by isolates of A. ochraceus on green coffee beans. Maximum mycelial growth was found at 30 degrees C and 0.95 to 0.99 a(w). A marked decrease in growth rate was observed when temperature and a(w) were reduced. At 0.80 a(w), mycelial growth occurred only at 30 and 20 degrees C for one isolate. Maximum OTA production was found at 20 degrees C and 0.99 a(w). At 10 degrees C, OTA was not produced, regardless of a(w). Similarly, no OTA was detected at 0.80 a(w). OTA production ranged from the limit of detection (40 ng g(-1) of green coffee) to 17,000 ng g(-1) of green coffee. Significant intraspecific differences in mycelial growth and OTA production were found. Primary data for lag phases prior to mycelial growth under the influence of temperature and a(w) were modelled by multiple linear regression, and the response surface plots were obtained.     

Impact of water activity and temperature on growth and ochratoxin A production of two Aspergillus carbonarius isolates from wine grapes in Greece

The aim of this study was to determine the effects of water activity (a(w); 0.85 to 0.98) and temperature (10 to 40 degrees C) on the radial growth rate and ochratoxin A (OA) production of two Aspergillus carbonarius isolates in vitro. The isolates were obtained from wine grapes cultivated in Greece, and the trial was conducted with a synthetic grape juice medium similar in composition to grapes between veraison (beginning of color change) and ripeness. Fungal growth and OA production data were collected for 55 days. Response surface curves and cardinal values for a(w) and temperature were obtained using multiple regression analysis. The lag phase lasted from less than 1 to 10 days. Both isolates grew optimally at 30 to 35 degrees C and 0.96 a(w), but maximum OA production occurred under suboptimal growth conditions (15 to 20 degrees C and 0.93 to 0.96 a(w)). Growth also was observed at 0.85 a(w) and 25 degrees C, however at this same a(w) the fungus failed to produce mycelium at any other temperatures tested. The isolates produced OA at 15 to 30 degrees C and 0.90 to 0.98 a(w). Maximum OA production was detected after 25 days of incubation at 20 degrees C and 0.96 a(w) and was 3.14 and 2.67 microg g(-1), respectively, for the two strains. The isolated strains used in this study were more xerotolerant than others from the Mediterranean basin. These data will allow producers to identify and thus monitor critical environmental conditions effectively in wine grapes. These data also increase the knowledge base concerning the ability of A. carbonarius to grow and produce toxin under different ecological conditions and can contribute to the development of secondary models for the prediction and risk assessment of OA in wine production.     

不同发芽条件对大麦中植酸含量及植酸酶活力的影响

大麦发芽过程中,发芽温度、浸麦水pH值及浸麦水中金属离子的种类和含量是影响大麦中植酸含量及植酸酶活力的重要因素。实验发现,当发芽温度为16℃时,植酸酶较高温和低温发芽更早的被激活,其活力达到4.032 8U/g(绝干),更加有利于植酸的分解;在浸麦水为中性条件下浸麦,植酸酶活力上升较快,植酸含量迅速下降;在浸麦期间添加Ca2+、Mg2和Fe3+等金属离子对植酸酶的活力有较大抑制作用。     

Modelling the effect of temperature and water activity of Aspergillus flavus isolates from corn

The aim of this study was to model the effects of temperature (10-40 °C) and a(w) (0.80-0.98), in two media (Czapek yeast agar: CYA; corn extract medium: CEM) on the growth rates and growth boundaries (growth-no growth interface) of three strains of A. flavus isolated from corn in Argentina. Both kinetic and probability models were applied to colony growth data. The growth rates obtained in CYA were significantly (p<0.05) greater than those obtained in CEM medium. No significant differences (p<0.05) were observed among the three isolates. The growth rate data showed a good fit to the Rosso cardinal models combined with the gamma-concept with R(2)=0.98-0.99 and RMSE=0.60-0.78, depending on media and isolates. The probability model allowed prediction of safe storage (p of growth <0.01) for one month for moist maize (e.g. 0.90 a(w)) provided temperature is under 15 °C, or for dry maize (e.g. 0.80 a(w)) provided temperature is under 27 °C. Storage at <0.77 a(w) would be safe regardless of the storage temperature. Probability models allow evaluation of the risk of fungal contamination in the process of storage, so the results obtained in this study may be useful for application in systems of food safety management.