Effect of sublethal concentrations of propionic or butyric acid on growth and aflatoxin production by Aspergillus flavus

Propionic or butyric acid was added at sublethal doses (0.1–2 mg/ml) to a growth medium supporting growth of Aspergillus flavus and subsequent aflatoxin production. A reduction in growth and aflatoxin production occurred when the acids were added at the time of inoculation. Addition of the acids to cultures at different times resulted in little effect on growth but production of aflatoxin after 12 days was reduced with earlier time of application for both propionic and butyric acid. When the acids were added to rough rice with a moisture content of 21% and inoculated with A. flavus fungal growth and aflatoxin production were reduced relative to non-inoculated controls. Early application of acids resulted in lower yields of aflatoxin.     

Inhibition of growth and aflatoxin production of Aspergillus parasiticus by citrus oils

Summary Aspergillus parasiticus was inoculated into grapefruit juice and a glucose-yeast extract medium; both contained 500–7000 ppm of citrus oils that were incorporated into the media by sonication. Orange and lemon oil were more inhibitory to mold growth and aflatoxin production than was d-limonene, the main constituent of the two peel oils. After 7 days at 28° C, 2000 ppm of lemon and 3000 ppm of orange oil in grapefruit juice afforded maximum suppression of mold growth and toxin formation. When the glucose-yeast extract medium was used, 3000 ppm of either oil were needed to achieve the same result. After 4 days at 28° C, orange oil at 3500 ppm in either medium markedly inhibited mold growth (as evidenced by dry weight of mold mycelium) and aflatoxin production (only 14 and 1% of the amount normally produced in the juice and artificial medium, respectively). Higher concentrations of orange oil further reduced mold growth and aflatoxin production and also delayed the onset of sporulation, if it occurred. Although aflatoxin was detected in all samples, only 0.2 to 0.5% of the amount found in controls (without the citrus oil) was present when the medium contained 7000 ppm orange oil. The mold consistently grew, albeit very poorly, on the glass at the liquid-atmosphere interface even when the substrate contained a large amount of citrus oil.

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Hemmung des Wachstums und der Aflatoxinproduktion von Aspergillus parasiticus durch Orangenöl, Citronenöl und d-Limonen

Zusammenfassung Pampelmusensaft und ein Glucose-Hefeextrakt-Medium wurden beide mit 500–7000 ppm Orangenöl, Citronenöl oder d-Limonen angereichert und dann mitAspergillus parasiticus beimpft. Wachstum und Aflatoxinproduktion des Pilzes wurden stärker durch die Öle als durch d-Limonen gehemmt, obwohl dieser der Hauptbestandteil der beiden Öle ist. 2000 ppm Citronenöl bzw. 3000 ppm Orangenöl in Pampelmusensaft genügten zur starken Hemmung der Wachstums- and der Aflatoxinproduktion vonA. parasiticus während 7 Tage bei 28° C. Wenn Glucose-Hefeextrakt als Nährboden diente, dann wiesen bei 3000 ppm beide Öle gleiche Hemmung auf. Wenn beide Nahrboden nur 4 Tage bei 28° C gehalten wurden, dann waren 3500 ppm Orangendl notwendig, um Wachstum and Aflatoxinproduktion zu hemmen. Pampelmusensaft mit einem Orangenöl-Gehalt von 3500 ppm enthielt nur 14% der Aflatoxin-Menge des beimpften Saftes ohne Öl. Das Medium mit Glucose, Hefeextrakt und Orangendl hatte nur 1 % des Aflatoxin-Gehaltes der Kontrolle. Höhere Konzentrationen von Orangenöl hemmten noch stärker und verzögerten den Beginn der Konidienbildung. Wenn das Medium 7000 ppm Orangenöl enthielt, dann konnte nur geringes Pilzwachstum und Aflatoxinproduktion (0,2–0,5% der Kontrolle) beobachtet wurden; das minimale Wachstum des Pilzes geschah an der Grenzfläche Nährboden und Atmosphare.

     

Antifungal properties and inhibitory effects upon aflatoxin production by Zingiber officinale essential oil in Aspergillus flavus

In this study, the efficacy of ginger (Zingiber officinale Roscoe) essential oil (GEO) in reducing A. flavus growth and aflatoxin production was investigated. Gas chromatography coupled to mass spectrometry and nuclear magnetic resonance spectroscopy showed that the major components of GEO were α‐zingiberene (23.85%) and geranial (14.16%). Mycelial growth of Aspergillus flavus was reduced significantly at a GEO concentration of 150 μg mL^?1, and complete inhibition of conidial germination was observed at a concentration of 10 μg mL^?1. Statistically significant inhibition of ergosterol biosynthesis was detected at a GEO concentration of 10 μg mL^?1. GEO was capable of fully inhibiting aflatoxin production by A. flavus at a concentration of 15 μg mL^?1. The results suggest that low concentrations of GEO are capable of inhibiting aflatoxin production; such ability could be valuable in the upcoming future for agricultural companies to better control aflatoxigenic fungi in agricultural products.     

Inhibitory effect of phenolics extracted from sorghum genotypes on Aspergillus parasiticus (NRRL 2999) growth and aflatoxin production

The inhibitory activity of bioactive polyphenols present in six sorghum genotypes—two red (AON 486 and IS 620), two yellow (LPJ and IS 17779) and two white (SPV 86 and SPV 462) varieties—on Aspergillus parasiticus (NRRL 2999) growth and aflatoxin production was evaluated. In the first experiment the production of aflatoxins in the six sorghum genotypes after removal of surface phenolics by acidic methanol treatment was studied and compared with that in untreated grains. Aflatoxin production was found to be fourfold higher in treated grains. The total phenols and bioactive polyphenols extracted by acidic methanol were quantified using the Folin–Denis method and the bovine serum albumin–benzidine conjugate procedure respectively. In the second experiment the effect of extracted sorghum phenolics under in vitro conditions on fungal growth and aflatoxin production was studied at two concentrations (0.01% and 0.1%) of phenolics. Extracted phenolics added to yeast extract sucrose (YES) medium at 0.1% concentration showed an inhibitory effect on aflatoxin production. At 0.01% phenolic concentration, aflatoxin production was minimal on day 3 after infection. At other time points the aflatoxin content was similar to that in the control. At 9 days after infection the fungal biomass in IS 620 was significantly lower than that in the control. At 0.1% phenolic concentration, aflatoxin production was minimal and the red genotype IS 620 showed maximum resistance. Fungal biomass was lowest at all growth stages in IS 620 as compared with the control. Polyphenol oxidase (PPO) activity was not detected in A. parasiticus grown on YES medium (control). PPO activity was not induced in A. parasiticus by the addition of phenolics to the liquid culture medium (no PPO activity was detected in the culture medium). The inhibitory activity of bioactive polyphenols could be attributed to the lack of PPO enzyme in this fungus. Copyright © 2007 Society of Chemical Industry     

Activity of essential oil and its major compound, 1,8-cineole, from Eucalyptus globulus Labill., against the storage fungi Aspergillus flavus Link and Aspergillus parasiticus Speare

The essential oil from leaves of Eucalyptus globulus obtained by hydrodistillation, as well as its major compound 1,8-cineole, identified by gas chromatography coupled with a mass selective detector, were evaluated for their effectiveness against the storage fungi Aspergillus flavus and Aspergillus parasiticus. The evaluation was performed by compound dissolution in yeast extract sucrose (YES) medium and exposure to headspace volatiles. Complete fungal growth inhibition of both species was achieved with the essential oil by contact and volatile assays. Volatile exposure showed total inhibition at the lower level tested of 500 μL. The 1,8-cineole tested alone showed partial inhibition only at the highest level of 1.3492 μL. Aflatoxin B₁ production was reduced in headspace volatile assays and partial inhibition was observed at the 200 μL dose of the essential oil.     

Effect of application of nontoxigenic strains of Aspergillus flavus and A. parasiticus on subsequent aflatoxin contamination of peanuts in storage

Experiments were conducted to determine the potential for biological control of aflatoxin contamination of peanuts during storage. Florunner peanuts were treated in field plots by applying competitive, nontoxigenic strains of Aspergillus flavus and A. parasiticus, at 76 and 67 days after planting in 1998 and 1999, respectively. After harvest, half the peanuts from both treated and control plots were sprayed with an aqueous conidial suspension containing the nontoxigenic strains; the other half of the peanuts from each group were not sprayed. The peanuts were then placed in separate compartments of a miniature warehouse. Therefore, storage treatments consisted of peanuts that were (1) not treated at all; (2) treated prior to storage only; (3) field-treated only; (4) treated both in the field and prior to storage. Peanuts were stored for 3-5 months under high temperature and relative humidity conditions designed to promote aflatoxin contamination. In 1998, peanuts were not contaminated with aflatoxins prior to storage. After storage, peanuts that were never treated with the competitive fungi contained an average of 78.0 ppb of aflatoxins. Peanuts not treated in the field but receiving the spray treatment before storage contained 48.8 ppb. Peanuts treated in the field only averaged 1.4 ppb, and peanuts treated both in the field and prior to storage contained 0.8 ppb. In 1999, peanuts suffered from late-season drought and were contaminated with aflatoxins at harvest, with controls averaging 516.8 ppb compared with 54.1 ppb in treated peanuts. After storage, non-field-treated peanuts averaged 9145.1 ppb compared with 374.2 ppb for peanuts that had been field-treated, a 95.9% reduction. Spraying of pods with the nontoxigenic strains postharvest but prior to storage provided no additional protection against aflatoxin contamination. Results demonstrated that field application of the nontoxigenic strains had a carry-over effect and reduced aflatoxin contamination that occurred in storage.     

Inhibitory effects of spice extracts on the growth of Aspergillus parasiticus NRRL2999 strain

 The inhibitory effects of 31 spices extracts on the growth of Aspergillus parasiticus NRRL 2999 strain were investigated in vitro. Of the 31 samples tested, thyme (wild), thyme (black), oregano and savory completely inhibited the growth of A.parasiticus at the 2% level in Czapek-Dox Agar and partially inhibited it at the 1% leves. It was also found that capers, parsley leaf, coriander, sumac, mustard (wild) and dill leaf markedly stimulated the mycelial growth. The effectiveness of the inhibitors followed the sequence: thyme (black)>oregano>savory>thyme (wild)>ajovan>rosemary. Received: 30 January 1998 / Revised version: 16 March 1998     

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.     

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.     

Inhibitory effect of the essential oil of Curcuma longa L. and curcumin on aflatoxin production by Aspergillus flavus Link

Aflatoxins are highly toxic, mutagenic, teratogenic and carcinogenic mycotoxins. Consumption of aflatoxin-contaminated food and commodities poses serious hazards to the health of humans and animals. Turmeric, Curcuma longa L., is a native plant of Southeast Asia and has antimicrobial, antioxidant and antifungal properties. This paper reports the antiaflatoxigenic activities of the essential oil of C. longa and curcumin. The medium tests were prepared with the oil of C. longa, and the curcumin standard at concentrations varied from 0.01% to 5.0%. All doses of the essential oil of the plant and the curcumin standard interfered with mycotoxin production. Both the essential oil and curcumin significantly inhibited the production of aflatoxins; the 0.5% level had a greater than 96% inhibitory effect. The levels of aflatoxin B₁ (AFB₁) production were 1.0 and 42.7 μg/mL, respectively, for the samples treated with the essential oil of C. longa L. and curcumin at a concentration of 0.5%.     

Sensitivity of four foodborne moulds to essential oils from turkish spices,herbs and citrus peel

The antifungal effects of 22 essential oils from Turkish spices, herbs and citrus peel on four foodborne moulds were evaluated for fungistatic and fungicidal activity. The most active oils were wild thyme (Thymus rariflorus L) and thyme (Thymus serpyllum L). Parsley (Penloselinum sativum Hoffin) and savory (Satureja hortensis L) oils were found to be the least active oils. Rhizopus sp displayed the greatest tolerance, and the most sensitive mould was Penicillium chrysogenum. Prolonged incubations reduced the fungistatic and fungicidal effects of oils. It was concluded that some oils may be useful as mould inhibitors at food additive levels.     

Modification of enzymatic and non-enzymatic in vitro oxidative defence system by bioaccessible phytonutrients of selected spices

The bioaccessibility and bioactivity of spice polyphenols using the in vitro model simulating gastro-intestinal conditions and intestinal absorption has been investigated.     

几种香辛料精油对产气荚膜梭菌的体外抑菌效果

研究白芷精油、黑胡椒精油、茴香精油、肉桂精油、姜精油和艾草精油对产气荚膜梭菌（Clostridium perfringens,C.perfringens）的抑制作用,筛选出对产气荚膜梭菌有抑菌效果的香辛料精油。采用牛津杯法和两倍稀释法分别判定不同香辛料精油对C.perfringens（ATCC13124、CICC22949、C₁和C₂）的抑菌圈大小和最小抑菌浓度（Minimum inhibitory concentration,MIC）。结果显示:肉桂精油对C.perfringens的抑菌圈直径最大,且对产气荚膜梭菌ATCC13124、CICC22949、C₁和C₂的MIC分别为2.75、2.75、5.5、2.75 mg/m L;艾草精油对C.perfringens的MIC分别为11、2.75、11、2.75 mg/m L;黑胡椒精油对C.perfringens的MIC分别为22、11、11、5.5 mg/m L;茴香精油对C.perfringens的MIC分别为11、5.5、11、2.75 mg/m L;姜精油对C.perfringens的MIC分别为88、22、22、22 mg/m L。结论:除白芷精油以外,其他5种香辛料精油对C.perfringens均有不同程度的抑菌作用,其中肉桂精油对C.perfringens的抑菌效果最佳。      

Effect of added citrus fibre and spice essential oils on quality characteristics and shelf-life of mortadella

The objective of this work was to study the effect of adding orange dietary fibre (1%), rosemary essential oil (0.02%) or thyme essential oil (0.02%) and the storage conditions on the quality characteristics and the shelf-life of mortadella, a bologna-type sausage. The moisture, fat, ash content and colour coordinates lightness (L∗) and yellowness (b∗) were affected by the fibre content. The treatments analysed lowered the levels of residual nitrite (57.56% and 57.61%) and the extent of lipid oxidation, while analysis of the samples revealed the presence of the flavonoids, hesperidin and narirutin. No enterobacteria or psychotropic bacteria were found in any of the treatments. The treated samples stored in vacuum packaging showed the lowest aerobic and lactic acid bacteria counts. Sensorially, the most appreciated sample was the one containing orange dietary fibre and rosemary essential oil, stored in vacuum packaging. Orange dietary fibre and spice essential oils could find a use in the food industry to improve the shelf-life of meat products.     

Effect of Capsicum carotenoids on growth and aflatoxins production by Aspergillus flavus isolated from paprika and chilli

The aim of this study was to determine the effect of a carotenoid mixture (Capsantal FS-30-NT), containing capsanthin and capsorubin, on growth and aflatoxins (AF) production of AF-producing Aspergillus flavus isolates.     

Chemical and aroma profiles of yuzu (Citrus junos) peel oils of different cultivars

The essential oils of six different yuzu cultivars, Kumon (KUM), Nagano (NAG), Yasu (YAS), Jimoto (JIM), Komatsu Sadao (KOS) and Komatsu Koichi (KOK), were extracted by cold-pressing method. A total of 69 compounds of the six samples were identified. Application of GC-olfactometry and aroma extraction dilution analysis technique in three-fold stepwise dilution of the neat oil for all samples indicated eight odourants with the highest flavour dilution (FD) values. Those were limonene, α-pinene, α- and β-phellandrene, myrcene, γ-terpinene, (E)-β-farnesene and linalool. ‘KOS’ was differentiated from the other oil samples by showing the highest number of components having yuzu-like odour notes and also from the PCA analysis of the FD-factor values. This is the first time the aroma characteristics of yuzu essential oils of specified cultivars were investigated.     

Application of essential oils in maize grain: impact on Aspergillus section Flavi growth parameters and aflatoxin accumulation

The antifungal activity of Pimpinella anisum L. (anise), Pëumus boldus Mol (boldus), Hedeoma multiflora Benth (mountain thyme), Syzygium aromaticum L. (clove), and Lippia turbinate var. integrifolia (griseb) (poleo) essential oils (EOs) against Aspergillus section Flavi was evaluated in sterile maize grain under different water activity (a_w) condition (0.982, 0.955, and 0.90). The effect of EOs added to maize grains on growth rate, lag phase, and aflatoxin B₁ (AFB₁) accumulation of Aspergillus section Flavi were evaluated at different water activity conditions. The five EOs analyzed have been shown to influence lag phase and growth rate. Their efficacy depended mainly on the essential oil concentrations and substrate water activity conditions. All EOs showed significant impact on AFB₁ accumulation. This effect was closely dependent on the water activity, concentration, and incubation periods. Important reduction of AFB₁ accumulation was observed in the majority of EO treatments at 11 days of incubation. Boldus, poleo, and mountain thyme EO completely inhibited AFB₁ at 2000 and 3000 μg g⁻¹. Inhibition of AFB₁ accumulation was also observed when aflatoxigenic isolates grew with different concentration of EOs during 35 days.     

Impact of volatile compounds generated by essential oils on Aspergillus section Flavi growth parameters and aflatoxin accumulation

BACKGROUND: The advantage of essential oils is their bioactivity in the vapour phase, a characteristic that makes them attractive as possible fumigants for stored grain protection. In this study the antifungal and antiaflatoxigenic effects of the volatile fractions of five essential oils (EOs) were evaluated by vapour contact on Aspergillus section Flavi isolates. RESULTS: In maize meal extract agar the volatile fractions of Pimpinella anisum L. (anise), Pëumus boldus Mol. (boldus), Hedeoma multiflora Benth. (mountain thyme), Lippia turbinata var. integrifolia (Griseb.) (poleo) and Syzygium aromaticum L. (clove) were able to decrease the growth rate and lag phase of aflatoxigenic isolates. Boldus EO showed the best antifungal effect on Aspergillus section Flavi growth rate. In sterilised maize grains, boldus and poleo EOs showed antifungal effects on growth rate and aflatoxin accumulation. The volatile fraction of boldus EO completely inhibited the growth of isolates at water activity (a_w) levels of 0.955, 0.930 and 0.900, while poleo EO showed this effect only at the lower a_w levels (0.930 and 0.900). All aflatoxigenic isolates showed reduced total aflatoxin accumulation in the presence of boldus EO under all a_w conditions. CONCLUSION: These findings clearly indicate that the volatile fraction of boldus EO could be used to control aflatoxigenic fungi in stored maize. Copyright © 2009 Society of Chemical Industry     

The production of aroma by Aspergillus oryzae during the preparation of soy sauce koji

The rôle of koji in the preparation of soy sauce has been a subject of controversy. A study of the volatiles produced during koji preparation and a comparison with uninoculated bean-flour mixture shows that the growth and sporulation of Aspergillus oryzae is related to the production of volatiles. The volatiles present at the mycelial stage are different from the volatiles of sporulated koji. The ether extracts of the acidic homogenates were different from those of the neutral and alkaline homogenates.