香菇纤维素衍生物抑制黄曲霉菌产毒的研究

黄曲霉毒素具有诱导突变、抑制免疫和致癌作用。控制黄曲霉毒素污染一直是世界性难题,也是近年来的研究热点之一。实验研究了香菇纤维素衍生物GPX抑制黄曲霉菌产毒活性,初步探究了GPX抑制黄曲霉菌产毒的作用机制。结果表明,10、50μg/mL的GPX对黄曲霉菌产毒的抑制率分别达到70%、95%,而100、250、500、750、1000μg/mL的GPX对黄曲霉菌产毒的抑制率均可达到100%,抑制活性极其明显。GPX离体抗氧化能力很低,但是能够缓解黄曲霉菌丝的氧化胁迫。此外GPX促进了黄曲霉细胞内囊泡过早与大液泡的融合,该现象能够抑制黄曲霉毒素的产生。      

Potential of aflatoxin B1 production by Aspergillus flavus strains on commercially important food grains

In this study, we investigated the potential of aflatoxin B₁ (AFB1) production by five Aspergillus flavus strains previously isolated from sorghum grains on cereals (barley, maize, rice, wheat and sorghum), oilseeds (peanuts and sesame) and pulses (greengram and horsegram). Five strains of A. flavus were inoculated on all food grains and incubated at 25 °C for 7 days; AFB1 was extracted and estimated by enzyme‐linked immunosorbent assay. All A. flavus strains produced AFB1 on all food grains ranging from 245.4 to 15 645.2 μg kg^?1. Of the five strains tested, strain Af 003 produced the highest amount of AFB1 on all commodities ranging from 2245.2 to 15 645.2 μg kg^?1. Comparatively, the AFB1 accumulation was high on rice grains ranging from 3125.2 to 15 645.2 μg kg^?1, followed by peanuts ranging from 2206.2 to 12 466.5 μg kg^?1. Less AFB1 accumulation was observed in greengram and sesame seeds ranging from 645.8 to 2245.2 and 245.4 to 2890.6 μg kg^?1, respectively. Our results showed that all food grains tested are susceptible to A. flavus growth and subsequent AFB1 production.     

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.     

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.     

乳杆菌及不同pH值对黄曲霉生长与产毒的影响

为探讨植物乳杆菌及不同PH值对黄曲霉生长与产毒的影响,将植物乳杆菌与黄曲霉孢子同时加入到MRS肉汤和先厌氧培养植物乳杆菌后再加入黄曲霉孢子;以乳酸将LTB培养基的初始PH值调整为3．0,4．0和5．0,以PH值为6．8为对照组,在LTB培养基中接种黄曲霉孢子悬液。上述试验均在28℃培养15d,在培养的第3、6、9、12和15天测定培养液中的PH值,黄曲霉素素B1和黄曲霉菌丝重量。结果显示：植物乳杆菌可显抑制黄曲霉生长（P＜0．05）,同时检测不到黄曲霉毒素B1。随着LTB培养液中PH值的降低,黄曲霉毒素B1的量增加,在PH3．0组,差异有极显性（P＜0．01）,各组间黄曲霉素菌丝量未见显性差异,结果表明：植物乳杆菌可显抑制黄曲霉生长与产毒。在PH值3．0－5．0范围内,乳酸可以刺激黄曲霉毒素B1的产生。     

Influence of Listeria monocytogenes and environmental abiotic factors on growth parameters and aflatoxin B1 production by Aspergillus flavus

Aspergillus flavus growth is influenced by a variety of abiotic factors including water activity (a_W), pH, redox potential, relative humidity, temperature and availability of oxygen. Furthermore, biotic factors such as the presence in the same habitat of different microorganisms can develop ecological interactions between them that are positive, negative or indifferent for fungal growth. The aim of the present work was evaluate the effect of Listeria monocytogenes in interaction with a_W, pH and temperature on the lag phase, growth rate and AFB₁ production of A. flavus. Ecophysiological in vitro studies of A. flavus demonstrate that the co-occurrence with L. monocytogenes in culture medium and abiotic factors (a_W, pH and temperature) influence the growth of the fungal strains and the production of AFB₁. The presence of L. monocytogenes generally decreased fungal growth and increased significantly aflatoxin production. These studies are essential in the prediction of contamination risk of food with these microorganisms or their toxins in interaction with several environmental parameters.     

含水量对玉米粉贮藏期黄曲霉生长及黄曲霉毒素B1积累的影响

目的 研究含水量对玉米粉贮藏过程中黄曲霉生长及黄曲霉毒素B1积累的影响。方法 将初始水分含量10.42%、13.26%、16.35%、19.88%的玉米粉在28 ℃, 相对湿度75%的模拟环境中储藏30 d; 测定贮藏期间玉米粉含水量、黄曲霉生长量及黄曲霉毒素B1的积累量。结果 贮藏期间玉米粉中含水量呈现先上升后下降的变化趋势, 黄曲霉生长量及黄曲霉毒素B1的积累量随贮藏时间的延迟先快速升高, 后趋于平稳。初始含水量较高的玉米粉贮藏期间含水量、黄曲霉生长量及黄曲霉毒素B1含量始终处于较高的水平。相关性分析表明, 玉米粉含水量、黄曲霉生长量和毒素积累量之间均呈极显著的正相关(P<0.01)。结论 含水量是影响玉米粉贮藏过程中霉菌生长和毒素积累的关键因素, 降低玉米粉初始含水量可以有效减轻贮藏期间真菌危害。     

Effect of Soybean Volatile Compounds on Aspergillus flavus Growth and Aflatoxin Production

ABSTRACT:  Soybean homogenates produced volatile compounds upon exposure to lipase. These induced volatiles were identified by SPME. Seventeen volatile compounds identified by SPME were chosen for determination of their ability to inhibit Aspergillus flavus growth and aflatoxin B₁ (AFB1) production in a solid media assay. These volatiles included aldehydes, alcohols, ketones, and furans. Of the tested compounds, the aldehydes showed the greatest inhibition of fungal growth and AFB1 production. These compounds inhibited up to 100% of the observed growth and AFB1 production as compared to the controls. The greatest activity by the aldehydes to disrupt growth was ranked as follows: 2,4 hexadienal > benzaldehyde > 2-octenal > ( E )-2-heptenal > octanal > ( E )-2-hexenal > nonanal > hexanal. The greatest activity by the aldehydes to reduce AFB1 was ranked as follows: ( E )-2-hexenal > 2,4 hexadienal > ( E )-2-heptenal > hexanal > nonanal. ( E )-2-hexenal and ( E )-2-heptenal were tested further in an A. flavus -inoculated corn kernel assay. Both compounds prevented colonization by A. flavus and eliminated AFB1 production when exposed to compound volumes < 10 μL as also shown in the solid media assay. The results suggest that soybeans react to lipase by production of potent antifungal volatiles.     

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.     

植物源天然产物对黄曲霉和黄曲霉毒素B_1的抑制作用研究进展

黄曲霉毒素B1(aflatoxin B1,AFB1)是化学物质中致癌性最强的一种,对人和动物的健康具有极大的威胁,尤其是对肝脏的损害。AFB1是谷物和粮油中最为多见的真菌毒素,危害性也最强,长期低剂量的摄入AFB1除了会导致肝脏损伤和诱发肝癌之外,也会损害人体的其他器官。因此,能够有效防护人体免遭AFB1损伤的功能食品和新型保健药物已成为研究重点。近年来,天然产物因其具有生物活性显著、副作用小、多靶点等优点而备受关注,多种植物源天然产物已被报道能够抑制黄曲霉的生长产毒和AFB1的毒性作用。该文主要综述了植物源天然产物对黄曲霉生长产毒和AFB1毒性作用的抑制作用以及其相关抑制机制,为进一步开展AFB1毒性损伤的防护研究工作,开发能降低AFB1对人和动物的毒性损伤的新型保健药物和功能食品提供参考。     

Development of a predictive model describing the growth of Yersinia enterocolitica in Camembert‐type cheese

Behaviour of Yersinia enterocolitica in mould‐ripened Camembert‐type cheese during storage at temperature range 3–15 °C was evaluated and mathematically described. The Baranyi and Gompertz models were adjusted to the results of the study to calculate the growth rate (GR) and lag time (LT) for Y. enterocolitica at each temperature. Goodness of fit was assessed by calculating the Akaike information criterion (AIC) and mean square error (MSE). Square root models were constructed which described the relations between GR, LT and applied storage temperature. The secondary models were mathematically validated based on the results generated by ComBase Predictor. Moreover, generated models were validated using external, independent data from ComBase database. Based on this, it was found that the square root models of Ratkowsky constructed on GR that were determined based on the Baranyi and Roberts model most accurately described the behaviour of Y. enterocolitica in Camembert‐type cheese during storage under refrigerated conditions.     

Assessment of azole fungicides as a tool to control growth of Aspergillus flavus and aflatoxin B1 and B2 production in maize

ABSTRACT

Aspergillus flavus is a highly aflatoxin (AF)-producing species infecting maize and other crops. It is dominant in tropical regions, but it is also considered an emerging problem associated with climate change in Europe. The aim of this study was to assess the efficacy of azole fungicides (prochloraz, tebuconazole and a 2:1 (w/w) mixture of prochloraz plus tebuconazole) to control the growth of A. flavus and AF production in yeast-extract–sucrose (YES) agar and in maize kernels under different water activities (a_w) and temperatures. Aflatoxins B₁ and B₂ were determined by LC with fluorescence detection and post-column derivatisation of AFB₁. In YES medium and maize grains inoculated with conidia of A. flavus, the growth rate (GR) of the fungus and AFB₁ and AFB₂ production were significantly influenced by temperature and treatment. In YES medium and maize kernels, optimal temperatures for GR and AF production were 37 and 25°C, respectively. In maize kernels, spore germination was not detected at the combination 37ºC/0.95 a_w; however, under these conditions germination was found in YES medium. All fungicides were more effective at 0.99 than 0.95 a_w, and at 37 than 25ºC. Fungicides effectiveness was prochloraz > prochloraz plus tebuconazole (2:1) > tebuconazole. AFs were not detected in cultures containing the highest fungicide doses, and only very low AF levels were found in cultures containing 0.1 mg l^–1 prochloraz or 5.0 mg l^–1 tebuconazole. Azoles proved to be highly efficient in reducing A. flavus growth and AF production, although stimulation of AF production was found under particular conditions and low-dosage treatments. Maize kernels were a more favourable substrate for AF biosynthesis than YES medium. This paper is the first comparative study on the effects of different azole formulations against A. flavus and AF production in a semi-synthetic medium and in maize grain under different environmental conditions.     

Correlation of growth and aflatoxin production by Aspergillus flavus with some essential metals in gamma irradiated crushed corn

The effects of gamma irradiation and some essential metals on growth and aflatoxin B1 production by Aspergillus flavus in crushed corn were investigated. The production of aflatoxin by A. flavus was influenced by the addition of zinc, copper or iron and the effect gradually decreased with increasing metal concentration from 0 to 300 ppm. A. flavus grew and depleted zinc, copper and iron at initial concentration of 100, 200 or 300 ppm. Presence of 100 ppm zinc, copper or iron plus gamma irradiation (0.5, 1.0, 2.0 kGy) enhanced the growth of A. flavus and the production of aflatoxin in contrast with irradiated samples alone. A. flavus was able to metabolize and deplete elements in all gamma-irradiated samples. These results suggest that stricter control of element levels in gamma irradiated grains could control aflatoxin contamination.     

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     

Effect of sake lees on cheese components in cheese ripened by Aspergillus oryzae and lactic acid bacteria

More than 2,000 varieties of cheese currently exist in the world, and cheese manufacture continues to flourish. To develop the cheese ripening process, additional ingredients are used during cheese production. In this study, the effect of sake lees as an additional ingredient on the fermentation of cheese using Aspergillus oryzae (koji mold), known as koji cheese, was investigated. Aspergillus oryzae is used in the fermentation of Japanese traditional foods, such as sake and soy sauce, given its strong enzymatic activities, as well as in cheese production (i.e., koji cheese). Sake lees, a by-product of the fermentation of rice with A. oryzae and yeasts in the sake brewing process, contains various metabolites, such as amino acids. Here, supplementation with sake lees enhanced the activities of lactic acid bacteria and affected the color of the cheese. Metabolome analysis revealed that sake lees altered the balance of carbohydrates and fatty acids in the cheese. Remarkably, supplementation with sake lees enhanced the production of umami-enhancing γ-glutamyl (kokumi-active) peptides. This study suggests that a new type of cheese can be produced using A. oryzae and sake lees, and information on the synergistic effects of A. oryzae and sake lees will aid the development of cheese production.     

Recovery and differentiation of long ripened cheese microflora through a new cheese-based cultural medium

A partial picture of the typical microflora of PDO Parmigiano Reggiano cheese was achieved by studying the cultivability of lactic acid bacteria associated with its manufacturing and ripening. A comprehensive sampling design allowed for the analysis of the cheese microflora during its production over 20 months of ripening. An innovative cheese agar medium (CAM) was prepared after testing 18 formulations all based on grated Parmigiano Reggiano ripened cheese. During cheese manufacturing and ripening, different samples were sampled and their microflora was recovered using CAM in comparison with other traditional media. Colonies which formed units from the different agar media tested were picked and isolated; the phylogenetic positions of 154 isolated strains were studied at level of species by 16S-rRNA gene sequencing. CAM seems to be able to recover the minority population coming from milk and whey starter, hardly estimable, during the first hours of production, on traditional media.     

Occurrence of aflatoxin M1 in randomly selected North African milk and cheese samples

Forty-nine samples of raw cow's milk and 20 samples of fresh white soft cheese were collected directly from 20 local dairy factories in the north-west of Libya and analysed for the presence of aflatoxin M₁ (AFM₁). The samples were analysed using a high-performance liquid chromatography technique for toxin detection and quantification. Thirty-five of the 49 milk samples (71.4%) showed AFM₁ levels between 0.03 and 3.13 ng ml^-1 milk. Multiple analyses of five milk samples free of AFM₁ artificially contaminated with concentrations of AFM₁ at 0.01, 0.05, 0.1, 1.0 and 3.0 ng ml^-1 showed average recoveries of 66.85, 72.41, 83.29, 97.94 and 98.25%, with coefficients of variations of 3.77, 4.11, 1.57, 1.29 and 0.54%, respectively. Fifteen of 20 white soft cheese samples (75.0%) showed the presence of AFM₁ in concentrations between 0.11 and 0.52 ng g^-1 of cheese. Multiple assays of five cheese samples free of AFM₁ spiked with different concentration of AFM₁ (0.1, 0.5, 1.0 and 3.0 ng g^-1) showed average recoveries of 63.23, 78.14, 83.29 and 88.68%, with coefficients of variation of 1.53, 9.90, 4.87 and 3.79%, respectively. The concentrations of AFM₁ were lower in the cheese products than in the raw milk samples.     

Effects of smoke produced from smoldering plants on the Aspergillus flavus growth and production of aflatoxin in pistachio

Aflatoxin (AF) contamination of the Iranian exporting pistachio has become a major problem in the last decades. In this study, the antifungal effects of smoke produced from the smoldering of several herbal plants were investigated. Four different ratios of plant weight/exposure time (5/5, 10/15, 15/30, and 20 g/45 min) were used from each plant material to smoke two isolates of Aspergillus flavus (A47 and A3), grown in potato dextrose agar (PDA). The results showed that the 20 g/45 min treatment using smoldering cinnamon bark, neem leaves, and clove flowers had efficient inhibitions of 100, 85, and 75%, respectively. Furthermore, the smoking of pistachio inoculated with a spore concentration of 1 × 10⁶ / ml using 30 g of cinnamon bark, neem leaves, and clove flowers for 75 min was capable of preventing the production of different types of AF, that is, B1, B2, G1, and G2 in the treated products. There is a good potential to smoke pistachio with these three herbal plants and prevent the production of AF in pistachio during the handling, storage, and transportation.     

Influence of gamma-irradiation and maize lipids on the production of aflatoxin B1 by Aspergillus flavus

The effect of gamma-irradiation and maize lipids on aflatoxin B1 production by Aspergillus flavus artificially inoculated into sterilized maize at reduced water activity (aw 0.84) was investigated. By increasing the irradiation doses the total viable population of A. flavus decreased and the fungus was completely inhibited at 3.0 kGy. The amounts of aflatoxin B1 were enhanced at irradiation dose levels 1.0 and 1.5 kGy in both full-fat maize (FM) and defatted maize (DM) media and no aflatoxin B1 production at 3.0 kGy gamma-irradiation over 45 days of storage was observed. The level in free lipids of FM decreased gradually, whereas free fatty acid values and fungal lipase activity increased markedly by increasing the storage periods. The free fatty acid values decreased by increasing the irradiation dose levels and there was a significant enhancement of fungal lipase activity at doses of 1.0 and 1.50 kGy. The ability of A. flavus to grow at aw 0.84 and produce aflatoxin B1 is related to the lipid composition of maize. The enhancement of aflatoxin B1 at low doses was correlated to the enhancement of fungal lipase activity.