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.     

Influence of white light,near-UV irradiation and other environmental conditions on production of aflatoxin B1 by Aspergillus flavus and ochratoxin A by Aspergillus ochraceus

The effects of illumination, near-ultraviolet, incubation temperature pH and some minor elements on the growth rate and production of aflatoxin B₁ by A. flavus and ochratoxin A by A. ochraceus were investigated. Aflatoxin B₁ and ochratoxin A production was considerably higher in the light than in the dark. The greatest aflatoxin B₁ and ochratoxin A production was occurred after 11 days of fermentation with light- and dark-grown cultures at 25 °C. The mycelial dry weight was also greater in the light than in the dark for both A. flavus and A. ochraceus. Exposure of conidia to near-UV irradiation increased mycelial dry weight and mycotoxins by both fungi more than white light. The greatest aflatoxin B₁ and ochratoxin A was at 25 °C with UV-grown culture (24 h exposure) producing a mean of 400 and 260 μg/50 ml of medium, respectively. The maximum aflatoxin B₁ and ochratoxin A yield was obtained at pH 5.5 and with increasing the initial pH to near neutrality, both mycotoxins yield decreased. Iron, cupper and zinc were observed to stimulate aflatoxin B₁ and ochratoxin A production and enhanced the growth rate of both A. flavus and A. ochraceus.     

Effect of γ‐irradiation on the biophysical and morphological properties of corn

The effects of γ‐irradiation on the biophysical and morphological properties of corn plants were investigated. The irradiation doses were 0, 1, 1.5, 2.5, 5, and 10 krad. Corn grains exposed to 1.5 and 2.5 krad showed highly significant changes in all growth parameters. Fluorescence and light absorption spectra of chlorophyll attributed to different doses treatments of corn grains clearly confirmed the superiority of 1.5 krad irradiation dose in stimulating corn plants. Furthermore, the frequency dependence of the relative permittivity and the electric conductivity of the treated samples have been performed and discussed. The obtained results give another support via the biophysical properties for the 1.5 krad irradiation dose to be the most favorable one to improve the plant growth characteristics.     

Effect of gamma-irradiation on the biophysical and morphological properties of corn

The effects of gamma-irradiation on the biophysical and morphological properties of corn plants were investigated. The irradiation doses were 0, 1, 1.5, 2.5, 5, and 10 krad. Corn grains exposed to 1.5 and 2.5 krad showed highly significant changes in all growth parameters. Fluorescence and light absorption spectra of chlorophyll attributed to different doses treatments of corn grains clearly confirmed the superiority of 1.5 krad irradiation dose in stimulating corn plants. Furthermore, the frequency dependence of the relative permittivity and the electric conductivity of the treated samples have been performed and discussed. The obtained results give another support via the biophysical properties for the 1.5 krad irradiation dose to be the most favorable one to improve the plant growth characteristics.     

EFFECT OF GAMMA IRRADIATION ON THE POSTHARVEST PHYSIOLOGY OF FIVE BANANA VARIETIES GROWN IN INDIA

ABSTRACT— Gamma irradiation to 20–40 krad inhibits the ripening changes in preclimacteric bananas without affecting the fruit quality. Both fruit maturity at harvest and post-irradiation storage temperature markedly influence the response to irradiation. The optimum dose and the maximum tolerable dose varied among the five varieties screened. Ability of the banana fruit to withstand higher doses of gamma irradiation depends on the physiological status of the fruit at time of irradiation. Irrespective of varietal differences, irradiation of preclimacteric bananas to doses above 50 krad resulted in severe skin discoloration and fruit splitting. Irradiation under anoxia did not markedly reduce the radiation injury, suggesting that factors other than ozone formed during irradiation in air may contribute the radiation damage. Fruits on the climacteric could tolerate up to 200 krad but no effect on ripening rate was observed. Ethylene or 2,4-D could reverse irradiation-induced inhibition of ripening in bananas. Irradiation seems to decrease the sensitivity of banana fruit to the ripening action of exogenously added ethylene.     

Effect of gamma irradiation on aflatoxin production in barley

Barley was inoculated with conidia of Aspergillus parasiticus NRRL 2999, and at 0°C was left to equilibrate at three moisture levels: 17, 20 and 25%. Gamma irradiation at five doses, 0, 50, 100, 200 and 400 krad, was applied to the grain either soon after moisture equilibration (3 days after inoculation) or 10 days later (13 days after inoculation). Increasing the radiation dose resulted in decreasing aflatoxin formation, with one exception: 200 krad applied 13 days after inoculation on barley stored at a moisture level of 25% (100% relative humidity) and 25°C led to higher aflatoxin production than at 100 or 50 krad. For the control of aflatoxin contamination in barley, storage at low moisture content appears to be more practical and effective than irradiation.     

Inhibitory effect of Aspergillus niger on the growth of Aspergillus ochraceus and Aspergillus flavus,and on aflatoxin formation

Aspergillus ochraceus and A flavus were grown on synthetic media (SM) supplemented with 50 or 200 ml litre^?1 SM on which A niger had been grown previously ( ‘A niger medium’ = ANM). Controls included SM acidified to pH 6.0 or 4.4, SM diluted with 50 or 200 ml litre^?1 water, and diluted-acidified SM. For both fungi, higher growth inhibition was recorded on ANM-containing SM than in the controls. Aflatoxin formation was markedly inhibited on SM to which 20 ml litre^?1 ANM extract (in methanol/chloroform, 2:1 v) had been added, although the growth of A flavus on that medium was almost the same as that in the control. It is concluded that the inhibitory effect of A niger on the growth of fungi should not be attributed merely to pH reduction, but also, mainly, to metabolites produced by the fungus in the growth medium, even at early stages of its growth.     

Radiosensitivity of Tribolium destructor Uyttenboogaart (Coleoptera: Tenebrionidae)

The development of adult Tribolium destructor Uyttenboogaart from eggs and larvae was prevented by exposing these immature stages to 5 krad of gamma irradiation. However, some adults did emerge when pupae were exposed to 5–30 krad. Adult longevity was significantly reduced, and sterility of both sexes was effected by doses of 10 krad and above. Control of this species therefore appears feasible with the levels of irradiation currently approved by the U.S. Food and Drug Administration (20–50 krad) for treatment of stored-grain insects in wheat and wheat flour in the U.S.     

The production of ochratoxin A and citrinin in barley

The production of ochratoxin A by Aspergillus ochraceus and of ochratoxin A and citrinin by Penicillium viridicatum growing on previously sterilised barley for 200 days at 5, 10 and 20°C and a water activity of 0.85 is reported. A. ochraceus did not grow at 5°C, multiplied slowly at 10°C but did not produce toxin. At 20°C the organism multiplied more quickly and produced ochratoxin after 19 days, which slowly disappeared over the next 150 days. P. viridicatum grew slowly at 5°C but did not produce any toxin. It multiplied at 10°C and produced ochratoxin A which was only detectable during the period from 100 to 150 days. At 20°C both ochratoxin A and citrinin were produced. Ochratoxin A was detected after 10 days and was still present after 240 days, whereas citrinin was produced in large quantities between 118 and 129 days and then rapidly disappeared.     

Response of malathion-susceptible and resistant strains of Tribolium castaneum (Coleoptera: Tenebrionidae) to a combination of 60Co gamma irradiation and malathion

Survival of malathion-susceptible (S) and malathion-resistant (R) strains of Tribolium castaneum (Herbst) adults exposed to gamma irradiation at 5, 10, 20 or 30 krad, followed by a 5-h exposure to malathion at 1, 8, 15 or 22 days after irradiation, was assessed for up to 5 weeks after irradiation. Exposure of S strain adults to malathion, 1 or 8 days after irradiation at all doses hastened the onset of death. Irradiation at 10, 20 and 30 krad was the dominant cause of death in the R strain. There was some evidence that exposure of the R strain to malathion at 1 or 8 days after irradiation at 10 krad accelerated mortality 2 and 3 weeks after exposure to radiation. No progeny were produced by either the S or R strains irradiated at any dose. Irradiation of S and R strains did not influence their response to malathion.     

Mould spoilage and mycotoxin formation in grains as controlled by physical means

Low oxygen concentrations (less than 1%) and/or increased concentrations of CO2 or N2 have been found to be highly effective in preventing the development of mould on grain and in inhibiting selected mycotoxins, e.g. aflatoxins, ochratoxin, patulin, penicillic acid and T-2. However, the levels of CO2 needed to inhibit mould growth are much higher than those required for the inhibition of mycotoxin production. The degree of inhibition achieved by elevated CO2 concentrations is dependent on other environmental factors, such as relative humidity (RH) and temperature. Nevertheless, the biosynthetic pathways for mycotoxin production are merely blocked, but not damaged by high CO2 levels. Irradiation has been shown to destroy the conidia of moulds but the information concerning the effect of irradiation on mycotoxin formation seems to be contradictory. Aflatoxin production was increased in irradiated wheat grain, but decreased in barley and maize when the grain was irradiated prior to inoculation. The number of spores in the inoculum, grain condition, relative humidity and other environmental factors could all affect the results obtained. However, ochratoxin formation by Aspergillus ochraceus was consistently enhanced by irradiation of spores or mycelium.     

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.     

Suppression of ochratoxin biosynthesis by naturally occurring alkaloids

The effects of four alkaloids on the biosynthesis of ochratoxin A (OTA), ochratoxin B (OTB) and citrinin were examined on four OTA-producing aspergilli: Aspergillus auricomus, A. sclerotiorum and two isolates of A. alliaceus. Piperine and piperlongumine, natural alkaloids of Piper longum, significantly inhibited OTA production at 0.001% (w/v) for all aspergilli examined. Piperine and piperlongumine affected the polyketide synthesis step of OTA production and inhibited production of citrinin. Curcumin, a constituent of tumeric, completely inhibited mycelial growth of A. alliaceus isolate 791 at 0.1% (w/v) and decreased OTA production by ∼70% at 0.01% (w/v). Sesamin, a constituent of sesame oil, inhibited OTA and OTB production by 60 and 45%, respectively, at 0.1% (w/v), showing its effect was on chloroperoxidase and polyketide synthase activity. The potential advantage of these natural products to reduce ochratoxin contamination of agricultural commodities is discussed.     

Analysis of duplicate 24-hour diet samples for aflatoxin B1, aflatoxin M1 and ochratoxin A.

In the spring and autumn of 1994, a total diet study, in which 123 participants collected duplicates of their 24-hour diets, was carried out. The goal of this study was to determine the mass fractions of a number of analytes in these duplicate diets, so as to be able to establish oral daily intake values. After measurements were carried out for pesticides, PCBs, elements, sterols, nitrate and nitrite, and fatty acids, the duplicate diet study was concluded with analyses for aflatoxin M1, aflatoxin B1 and ochratoxin A. For this purpose a method of analysis was developed, that could simultaneously determine these mycotoxins at very low levels. The method involved chloroform extraction, liquid-liquid extraction, immunoaffinity cleanup and liquid chromatography. The method was supplemented with a procedure to confirm the identity of chromatographic peaks, assumed to represent aflatoxin M1, aflatoxin B1 and ochratoxin A. The method was in-house validated. Recoveries ranged from 68-74% for aflatoxin M1 (at spiking levels from 30-120 ng/kg, c.v. 7.6%), from 95-97% for aflatoxin B1 (at spiking levels from 50-200 ng/kg, c.v. 2.8%), and from 75-84% for ochratoxin A (at spiking levels from 150-600 ng/kg, c.v. 4.3%). Limits of quantitation (defined as signal/noise = 10) were estimated to be 24, 5 and 16 ng/kg lyophilised material for aflatoxin M1, aflatoxin B1 and ochratoxin A respectively. The newly developed method was used to analyse 123 samples of 24-hour diets. Aflatoxin M1 was detectable in 48% of the samples; the toxin contents remained below the limit of quantitation in all samples. Aflatoxin B1 could be detected in 42% of the samples; in 25% of the samples the levels were above the limit of quantitation. Ochratoxin A could be quantified in all samples. The analytical results were further processed to estimate levels of intake. Intake levels for the aflatoxins were very low, and could not reliably be established. The mean ochratoxin A intake was estimated to be 1.2 ng/kg body weight per day. This is well below the tolerable daily intake established by JECFA at 14 ng/kg body weight per day. The current dietary intake of ochratoxin A in the Netherlands is concluded to pose no appreciable health risk.     

Effect of gas barrier characteristics of films on aflatoxin production by Aspergillus flavus in peanuts packaged under modified atmosphere packaging (MAP) conditions

The effects of the gas barrier characteristics of three films (ASI, ASII and ASIII) and storage temperature on the growth of, and aflatoxin production by, Aspergillus flavus in peanuts packaged in air and under a modified gas atmosphere of CO₂:N₂ (65:35) were investigated. Mold growth was barely visible in air packaged peanuts using high-medium barrier films (ASI and ASII) and stored at 20°C with more extensive growth occurring in air packaged peanuts stored at 25 and 30°C. Extensive mycelial growth and sporulation occurred in all air packaged peanuts in a low barrier film (ASIII), especially at 30°C. Gas packaging inhibited mold growth in peanuts packaged in a high gas barrier film at 20°C. However, mold growth occurred in gas packaged peanuts packaged in film ASII at higher storage temperatures while extensive mycelial growth was observed in all peanuts packaged in film ASIII irrespective of storage temperature. Levels of aflatoxin greater than the regulatory limit of 20 ng/g were detected in all air packaged peanuts with the highest level of aflatoxin (76 ng/g) being detected in peanuts packaged in a high gas barrier film ASI. Aflatoxin production was inhibited in gas packaged peanuts using a high barrier film. However, higher levels of aflatoxin were detected in all gas packaged peanuts in medium-low gas barrier films (ASII and ASIII), particularly at higher storage temperatures. This study has shown that MAP using a CO₂:N₂ (65:35) gas mixture was effective in controlling aflatoxin production by A. flavus in peanuts to levels less than the regulatory limit of 20 ng/g. However, the antimycotic effect of low O₂-high CO₂ atmospheres is dependent on the gas barrier characteristics of the packaging films, especially at higher storage temperatures.     

Inhibitory effects of anethole and eugenol on the growth and toxin production of Aspergillus parasiticus

The antifungal and antiaflatoxigenic activity of anethole and eugenol which are active components of commonly used spices was studied against two strains of Aspergillus parasiticus. Anethole, up to concentration of 400 μg/ml where complete inhibition was observed, delayed growth and reduced mycelial weight but it showed a stimulative effect on the toxin production of both strains. At a concentration of 300 μg/ml, eugenol inhibited the growth of both strains; levels of eugenol below 200 μg/ml enhanced production of aflatoxin particularly by A. parasiticus NRRL 299.     

Influence of light on food relevant fungi with emphasis on ochratoxin producing species

The influence of light of varying wavelength on growth and ochratoxin A biosynthesis of Aspergillus carbonarius, A. niger, A. steynii and on Penicillium nordicum and P. verrucosum was analysed. For comparison the influence of light on various other food relevant fungi, including citrinin producers, was also tested. Generally the Aspergilli seem to be more resistant to light treatment than the Penicillia. Interestingly wavelengths from both sides of the spectrum, e. g. red (long wavelength, 627 nm) and blue (short wavelength 470-455 nm) had the strongest inhibitory effects on growth and ochratoxin A biosynthesis. Blue light generally had a stronger effect. Light of moderate wavelength, 590 to 530 nm, (yellow to green) had more a positive than a negative influence on growth or ochratoxin A biosynthesis compared to the control (dark incubation). The light effect on growth and ochratoxin A biosynthesis was dependent on the growth medium. In contrast to malt extract medium (MEA), YES medium, as an especially nutrient rich medium, had an attenuating effect on the reactivity against light. However the tendency of the response in both media was the same. Moreover, the light intensity strongly influences how the fungus reacts. Depending on the intensity and the resistance of the species a complete cessation of growth and/or inhibition of ochratoxin A biosynthesis could be achieved. Light irradiation has the opposite effect on ochratoxin A than citrinin, two mycotoxins which can be produced simultaneously in P. verrucosum. Citrinin was produced essentially under light conditions which inhibited ochratoxin A biosynthesis. The same was true for a derivative of ochratoxin, in particular a derivative of ochratoxin β in A. carbonarius. A. carbonarius produced high amounts of the ochratoxin β derivative under blue light when the production of ochratoxin A was ceased at the most inhibiting conditions used (MEA, royal blue light, 455 nm, 1700 lx). Light has a growth stalling but not inactivating effect on aerial mycelia. If a non-growing colony under light is shifted to the dark it immediately grows normally. However on spores blue light has a deactivating effect. After incubation of spores of P. verrucosum for 24 h under blue light up to 97% of the spores were no longer able to germinate. Again the spores of the Aspergilli were much more resistant.     

Reduction of aflatoxins by Korean soybean paste and its effect on cytotoxicity and reproductive toxicity--part 1. Inhibition of growth and aflatoxin production of Aspergillus parasiticus by Korean soybean paste (Doen-jang) and identification of the active component

The inhibitory effect of methanol extract of Korean soybean paste on the mold growth and aflatoxin production of a toxigenic strain of Aspergillus parasiticus ATCC 15517 was studied using different concentrations of the extract in yeast-extract sucrose broth. While inhibition in mold growth due to increasing the concentration of the extract was observed, the more remarkable effect was the inhibition of aflatoxin production. Reduction of mycelial weight as a result of addition of the extract was observed to range between 1.5 to 12.9% while reduction of aflatoxin production quantified by high-performance liquid chromatography ranged from 14.3 to 41.7%. Five percent of the extract significantly reduced aflatoxin production at the end of the incubation period (P < 0.05), although the effect on mycelial growth was less pronounced. This study indicates that soybean paste could also be an effective inhibitor of aflatoxin production even though mycelial growth may be permitted. The main active component identified by gas chromatography-mass spectroscopy was linoleic acid.     

通过辐射方法利用接枝共聚原理对棉人造丝和天然丝进行改性处理

在^60Co-γ射线照射下，甲基丙烯酸甲酯（MMA）以不同的浓度溶解于甲醇溶剂中，能够成功地接枝到棉、人造丝和天然丝纱线中。其中，天然丝中MMA的接枝量最高，达到35％；其次是人造丝22％；接枝量最低的是棉纱，仅为17％。在辐射总强度为500krad，辐射速率为200krad／h的时候，能产生最好的接枝效果。如果在MMA＋MeOH溶液（甲基丙烯酸甲酯与甲醇溶液）体系中加入少量的复合添加剂既可以提高接枝量，又能够赋予接枝纱线一定的抗拉强度，这表明功能复合添加剂在接枝反应中对增强纱线抗拉强度起到很好的作用。经过接枝共聚改性处理后，纱线的吸水率和平衡回潮率受到了主要影响。     

Glufosinate-ammonium reduces growth and aflatoxin B1 production by Aspergillus flavus

The herbicide glufosinate-ammonium (GA) [butanoic acid, 2-amino-4-(hydroxymethylphosphinyl)-ammonium salt] was tested at concentrations from 2 to 2,000 g GA per ml for activity against growth and aflatoxin B1 (AFB) production by the mycotoxigenic fungus Aspergillus flavus Link:Fr. The highest concentration (2,000 microg GA per ml) reduced colony diameter of A. flavus strain AF13 by 80%. AFB1 production was inhibited by 90% at this concentration. Reduction in mycelial dry weight and AFB1 production in response to GA application ranged from 17.2 to 97.1% and from 39.1 to 90.1%, respectively. Of four concentrations tested, 2 microg GA per ml was weakly inhibitory. In the kernel screening assay, AFB1 production was inhibited 60 to 91% when kernels were preimmersed or immersed 5 days after incubation in 200 microg GA per ml. Both concentrations (2 and 200 microg GA per ml) reduced seed germination by 25 to 50%. Results indicate that GA has an inhibitory effect on growth and AFB1 production by A. flavus.