Efficacy of water-dispersible formulations of biological control strains of Aspergillus flavus for aflatoxin management in corn

Field experiments were conducted in 2011 and 2012 to evaluate the efficacy of water-dispersible granule (WDG) formulations of biocontrol strains of Aspergillus flavus in controlling aflatoxin contamination of corn. In 2011, when aflatoxin was present at very high levels, there was no WDG treatment that could provide significant protection against aflatoxin contamination. The following year a new WDG formulation was tested that resulted in 100% reduction in aflatoxin in one field experiment and ≥ 49% reduction in all five WDG treatments with biocontrol strain 21882. Large sampling error, however, limited the resolution of various treatment effects. Corn samples were also subjected to microbial analysis to understand better the mechanisms of successful biocontrol. In the samples examined here, the size of the A. flavus population on the grain was associated with the amount of aflatoxin, but the toxigenic status of that population was a poor predictor of aflatoxin concentration.     

黄曲霉菌株的筛选和产毒能力鉴定

本研究从发霉粮食中分离出数株黄曲霉菌菌株,并进行了形态学和分子生物学鉴定。为了探究分离菌株与黄曲霉标准菌株之间产毒能力的差异,通过对分离菌株和黄曲霉标准菌株进行发酵培养和HPLC测定,分析确定产毒能力。结果表明黄曲霉菌株之间产毒能力差异巨大：黄曲霉菌株3.4408产毒量很高,黄曲霉菌株HDWH产毒量很低,黄曲霉菌株3.2572甚至不产生黄曲霉毒素;产生黄曲霉毒素菌株中部分黄曲霉菌株产生四种黄曲霉毒素AFB1、AFB2、AFG1、AFG2,黄曲霉菌株HDWS只产生黄曲霉毒素AFB1、AFB2。     

二氢杨梅素抑制黄曲霉菌生长的机制

本研究探讨了二氢杨梅素对黄曲霉的抗真菌活性和潜在的抗真菌机制。首先,通过抑菌实验证明,二氢杨梅素对黄曲霉孢子和菌丝的最小抑菌质量浓度（minimum inhibitory concentration,MIC）均为4 mg/mL。通过荧光增白剂（calcofluor white,CW）和碘化丙锭（propidium iodide,PI）染色实验证明,二氢杨梅素处理后黄曲霉细胞壁和细胞膜受损。与对照组相比,1/2 MIC和MIC组黄曲霉细胞内容物释放量（OD260 nm）分别增加了3.14 倍和5.93 倍,细胞外相对电导率和pH值均升高,MIC的二氢杨梅素对黄曲霉的呼吸抑制率达25.82%。这些结果表明,二氢杨梅素通过破坏细胞壁和细胞膜的完整性以及干扰呼吸代谢发挥抗菌活性。此外,二氢杨梅素在MIC时能够完全抑制黄曲霉在花生和玉米籽粒上的萌发,因此,二氢杨梅素可作为一种有效的抗真菌天然化合物应用于粮食及农产品储藏中。     

Relative populations and toxin production by Aspergillus flavus and Fusarium verticillioides in artificially inoculated corn at various stages of development under field conditions

The presence, development and production of mycotoxins by Aspergillus flavus and Fusarium verticillioides were studied in corn ears under field conditions after artificial contamination of corn silks. The planted area was divided into five treatments: T1, inoculated with A. flavus solution containing 1 × 10⁸ spores, ears covered; T2, inoculated with F. verticillioides solution containing 1 × 10⁸ spores, ears covered; T3, inoculated with F. verticillioides plus A. flavus solution containing 1 × 10⁸ spores of each, ears covered; T4, sprayed with sterile phosphate‐buffered saline, ears covered; T5, non‐sprayed silks, uncovered ears. Soil and air samples were also collected and analysed for the occurrence of fungi. Water activity, relative air humidity, rainfall and temperature were determined to assess the correlation between abiotic factors and the presence of fungi in the samples. Contamination with the inoculated fungus predominated in T1 and T2. In the other treatments, F. verticillioides was the most frequently isolated contaminant irrespective of treatment. Considering the production of mycotoxins, a positive relation between the production of fumonisins B₁ and B₂ and the frequency of F. verticillioides was statistically verified in all treatments. Copyright © 2007 Society of Chemical Industry     

Effect of γ‐irradiation on aflatoxin B1 production by Aspergillus flavus and chemical composition of three crop seeds

The effect of γ‐irradiation on aflatoxin B₁ production by Aspergillus flavus, and the chemical composition of some different crop seeds were investigated. A. flavus infected seeds behaved differently according to their principal constituents. A. flavus caused an increase in protein and decrease in lipids and carbohydrate contents of wheat, soyabean and fababean seeds. Growth of A. flavus and production of aflatoxin B₁ was inhibited at a dose level of 5 kGy. A. flavus utilizes carbohydrates of seeds for its growth and aflatoxin production. Crops were arranged, in descending order, according to aflatoxin produced in seeds as wheat > soyabean > fababean. There were no changes in chemical constituents of irradiated seeds, such as protein, lipids, and carbohydrates.     

Delivery systems for biological control agents to manage aflatoxin contamination of pre-harvest maize

While soil application of a competitive non-toxigenic Aspergillus flavus strains is successful in reducing aflatoxin contamination in certain crops, direct application to aerial reproductive structures could be more effective for maize. A sprayable, clay-based water-dispersible granule formulation was developed to deliver non-toxigenic A. flavus strain K49 directly to maize ears. The efficacy of the K49 water-dispersible granule in mitigating aflatoxin in maize (Zea mays L.) was evaluated. Field studies were conducted to compare K49 colonization and effectiveness in reducing aflatoxin contamination when applied either as a soil inoculant or as a directed spray in plots infested with toxigenic strain F3W4. Fifty percent of non-toxigenic A. flavus was recovered from non-treated controls and from plots soil inoculated with K49 on wheat. In spray treatments with formulated or unformulated K49 conidia, over 90% of A. flavus recovered was non-toxigenic. Soil-applied K49 reduced aflatoxin contamination by 65% and spray applications reduced contamination by 97%. These findings suggest direct spray application of non-toxigenic A. flavus strains may be better than soil inoculation at controlling maize aflatoxin contamination and that a water-dispersible granule is a viable delivery system for maintaining viability and efficacy of the biological control agent, K49.     

植酸对黄曲霉菌(Aspergillus flavus)糖化力的影响

该文通过添加不同浓度的植酸于黄曲霉菌(Aspergillus flavus)培养基中，用不同添加方式，于不同生长时刻测定黄曲霉菌糖化力，研究植酸对黄曲霉菌糖化力的影响。结果表明加入植酸后黄曲霉菌糖化力均有很大提高，可为工业生产带来可观的经济效益。植酸最佳作用浓度范围是0．20％～0．25％，发酵24h糖化力最大可提高44．32％。     

Biodiversity of Aspergillus section Flavi in the United States: A review

Fungi belonging to Aspergillus section Flavi are of great economic importance in the United States due to their ability to produce toxic and carcinogenic aflatoxins in agricultural commodities. Development of control strategies against A. flavus and A. parasiticus, the major aflatoxin-producing species, is dependent upon a basic understanding of their diversity in agricultural ecosystems. This review summarizes our current knowledge of species and population diversity in the United States in relation to morphology, mycotoxin production and genetic characters. The high genetic diversity in populations of aflatoxigenic fungi is a reflection of their versatile habits in nature, which include saprotrophic colonization of plant debris in soil and parasitism of seeds and grain. Genetic variation within populations may originate from a cryptic sexual state. The advent of intensive monoculture agriculture not only increases population size but also may introduce positive selective pressure for aflatoxin production due to its link with pathogenicity in crops. Important goals in population research are to determine how section Flavi diversity in agricultural ecosystems is changing and to measure the direction of this evolution.     

Use of PCR for Detecting Aspergillus flavus in Maize Treated by Gamma Radiation Process

The aim of this study was to verify the effects of gamma radiation process on the fungal DNA and the application of PCR in the detection of Aspergillus flavus in irradiated maize grains. The samples were inoculated with a toxigenic strain and incubated under controlled conditions of relative humidity, water activity, and temperature for 15 days. After incubation, the samples were treated with gamma radiation with doses of 5 and 10 kGy and individually analyzed. The use of PCR technique showed the presence of DNA bands of Aspergillus flavus in all irradiated samples that showed no fungal growth in agar medium.     

不同基因型玉米籽粒发育过程中糖代谢相关酶活性的比较

对高淀粉玉米与低淀粉玉米籽粒发育过程中蔗糖代谢和淀粉合成相关酶活性的变化动态进行了比较研究。结果表明,费玉3号籽粒支链淀粉、直链淀粉及总淀粉积累速率高于豫玉22;费玉3号籽粒支链淀粉、直链淀粉及总淀粉含量大于豫玉22;豫玉22的高峰黏度、低谷黏度、最终黏度及胶凝值均显著高于费玉3号,其崩解值和糊化温度低于费玉3号;豫玉22的可溶性糖含量高于费玉3号;费玉3号的腺苷二磷酸葡萄糖焦磷酸化酶(ADPG-PPase)活性、尿苷二磷酸葡萄糖焦磷酸化酶(UDPG-Ppase)活性、可溶性淀粉合成酶(SSS)活性、束缚态淀粉合成酶(GBSS)活性高于豫玉22。     

植酸对黄曲霉菌糖化酶活力的影响

通过添加不同浓度的植酸于黄曲霉菌（Aspergillus flavus)02-6-58培养基中，于不同生长时间、不同添加方式测定黄曲霉菌糖化酶活力，研究植酸对黄曲霉菌糖化酶活力的影响。结果表明：(1)加入植酸后其糖化酶活力均有很大提高。其最佳作用浓度范围是0．20％-0．25％，最大可提高44．32％；(2)在黄曲霉菌生长麸曲中添加植酸比在连续培养3代的斜面培养基中加入植酸糖化酶活力提高幅度更大；(3)比较黄曲霉菌生长曲线及产酶曲线，加入植酸后，其对数生长期提前3～4h，平衡期延长，从而提前进入产酶高峰期。     

不同品种玉米挤压膨化特性研究

以黄淮海平原玉米主产区53个玉米品种为材料,以德国布拉本德食品仪器公司DSE-25型双螺杆挤压膨化实验室工作站为膨化设备,系统研究了不同玉米品种籽粒的挤压膨化特性。结果表明,在相同挤压膨化工艺条件下(水分为17%,五区温度180℃,螺杆转速120 r.m in-1,喂料速度16 r.m in-1),不同玉米品种籽粒挤压膨化物的产品特性差异较大,而挤压膨化时的系统参数差异较小。与夏玉米品种相比,春玉米品种籽粒挤压膨化物具有较高的径向膨化率、吸水性指数和产量以及较低的水溶性指数和机械能耗。玉米品种挤压膨化物的径向膨化率平均为1.53,容积密度为0.196g.mL-1,水溶性指数为38.44%、吸水性指数为430.00%,扭矩为151.33 N.m、四区压力为26.66bar、五区压力为10.77bar,产量为2.27 kg.h-1,机械能耗为841.4 W.h.kg-1。玉米籽粒的挤压膨化特性与其理化特性有关。     

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。     

Fumonisins and their analogues in contaminated corn and its processed foods – a review

ABSTRACT

One of the food security problems faced worldwide is the occurrence of mycotoxins in grains and their foods. Fumonisins (FBs) are mycotoxins which are prevalent in corn (Zea mays L.) and its based foods. Their intake and exposure have been epidemiologically and inconclusively associated with oesophageal cancer and neural tube defects in humans, and other harmful health effects in animals. The toxic effects of FBs can be acute or chronic and these metabolites bioaccumulate mainly in liver and kidney tissues. Among FBs, fumonisin B₁ (FB₁) is the most relevant moiety although the ‘hidden’ forms produced after food thermal processes are becoming relevant. Corn is the grain most susceptible to Fusarium and FBs contamination and the mould growth is affected both by abiotic and biotic factors during grain maturation and storage. Mould counts are mainly affected by the grain water activity, the environmental temperature during grain maturation and insect damage. The abiotic factors affected by climatic change patterns have increased their incidence in other regions of the world. Among FBs, the hidden forms are the most difficult to detect and quantify. Single or combined physical, chemical and biological methods are emerging to significantly reduce FBs in processed foods and therefore diminish their toxicological effects.     

Assessment for fungal,mycotoxin and insect spoilage in maize stored for human consumption in Zambia

BACKGROUND: Maize constitutes the main staple food and most important crop grown in Zambia. However, maize incurs considerable losses both in field and storage due to pathogens and insects. Some of the pathogens and resultant mycotoxins reduce the nutritional quality of the product. Mycotoxins are toxigenic fungal compounds that can cause cancer and suppress growth. In spite of this health hazard, there has been very little research to document their occurrence. Maize grains stored for human consumption were sampled from different agro‐ecosystems (forest, valley and plateau areas) of three agroecological zones (high, mid and low altitude). RESULTS: Several fungal genera were recovered among which Aspergillus flavus, A. niger, Fusarium verticillioides, F. solani, Rhizopus stolonifer and Penicillium spp. were prevalent. The weevil Sitophilus zeamais and the larger grain borer Prostephanus truncatus were the most damaging. Enzyme‐linked immunosorbent assay (ELISA) tests yielded fumonisins and aflatoxins ranging between 0.02 and 21.44 ppm, and 0.7 and 108.39 ppb in 96.4% and 21.4% of samples, respectively. Fumonisin was more pronounced in villages in forest areas whereas aflatoxin was highest in valley and forest areas in Zone II. CONCLUSION: Strategic interventions to curtail fungal, mycotoxin and insect contamination should be directed towards improved agronomic and post‐harvest practices of maize from fields to consumers. Copyright © 2009 Society of Chemical Industry     

黄玉米粗类黄酮提取工艺正交试验优化及其体外抗氧化活性

以黄玉米纪元1号为材料,采用单因素试验与三元二次正交试验,考察乙醇体积分数、料液比、浸提温度及浸提时间对粗类黄酮提取量的影响及黄玉米粗类黄酮清除1,1-二苯基-2-苦肼基（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基和•OH的能力。得出最优工艺参数为料液比1∶35.5（g/mL）、69 ℃浸提1.85 h,此条件下粗类黄酮提取量为85.88 mg/g。当粗类黄酮质量浓度为20 mg/mL时,对DPPH自由基和•OH的清除率分别为76.47%和32.62%,分别相当于芦丁标准品（0.1、0.2 mg/mL）清除能力的81%～84%和40%～50%。黄玉米粗类黄酮对DPPH自由基的清除率明显大于对• OH的清除率,且随质量浓度提高,清除率显著提高（P＜0.05）。     

Aspergillus flavus population isolated from soil of Argentina's peanut‐growing region. Sclerotia production and toxigenic profile

The Aspergillus flavus population was evaluated in the period 1998–2001 in soil samples from the peanut‐growing region in Argentina. A total of 369 A flavus isolates were examined for sclerotia, aflatoxin and cyclopiazonic acid production. The L phenotype was isolated in a higher percentage than the S phenotype and represented 59% of the total isolates. Statistical analysis showed significant differences between L, S and non‐sclerotial strains with regard to aflatoxin and cyclopiazonic acid production (p < 0.05). The S strains produced higher mycotoxin levels than the L and non‐sclerotial strains. About 10% of the S strains had an unusual pattern of mycotoxin production because they simultaneously produce aflatoxins B and G and CPA. The S_BG strains isolated in the present study have all morphological and microscopic characteristics of A flavus. These strains are of concern in food safety, as there is a higher probability of aflatoxin contamination in peanuts. Copyright © 2005 Society of Chemical Industry     

黄玉米粗类黄酮在牛肉丸保鲜中的应用

研究黄玉米粗类黄酮的不同添加量对牛肉丸在4℃贮藏期间的保鲜效果。将加工好的牛肉丸随机分为五组:不添加任何保鲜剂的空白对照组、加入0.02%丁基羟基茴香醚(Butylated Hydroxyanisole,BHA)的阳性对照组、分别加入0.02%、0.06%、0.10%黄玉米粗类黄酮的试验组,在4℃的贮藏条件下,定期检测各组牛肉丸的pH、硫代巴比妥酸反应物(Thiobarbituric Acid Reactive Substance,TBARS)、挥发性盐基总氮(total volatile basic nitrogen,TVB-N)值和菌落总数。结果表明:与空白组相比,BHA与黄玉米粗类黄酮均可以显著抑制贮藏期间牛肉丸中脂质氧化(P<0.05)、蛋白质氧化、微生物快速繁殖(P<0.05),延缓牛肉丸变质;黄玉米粗类黄酮处理组中,0.06%黄玉米粗类黄酮对牛肉丸的保鲜效果更佳;0.10%黄玉米粗类黄酮抑制牛肉丸菌落繁殖效果最佳。黄玉米粗类黄酮能有效抑制牛肉丸脂质和蛋白氧化以及微生物繁殖,延长货架期,有潜力作为天然抗氧化剂。     

2-Hydroxy-4-methoxybenzaldehyde inhibits the growth of Aspergillus flavus via damaging cell wall,cell membrane,manipulating respiration thus creating a promising antifungal effect on corn kernels

This study investigated the antifungal activity and the potential antifungal mechanisms of 2-hydroxy-4-methoxybenzaldehyde (HMB) against Aspergillus flavus (A. flavus). The minimum inhibitory concentration (MIC) of HMB in preventing spore germination was 70 μg mL⁻¹. HMB at MIC disrupted cell wall integrity by reducing the number of septa by 86.66% (P < 0.05) in mycelia and increased cell membrane permeability by about 14-fold (P < 0.05) evidenced by propidium iodide (PI) staining. Furthermore, HMB at MIC inhibited respiration by 33.33%. These results revealed that the antifungal activity of HMB against A. flavus could be attributed to the damaged cell wall integrity, cell membrane permeability and respiration metabolism. What’s more, A. flavus was completely restrained in corn kernels due to HMB. Therefore, HMB could be applied as an effective antifungal agent.