Growth of spoilage mould and aflatoxin B1 production in naturally contaminated or artificially inoculated maize as influenced by moisture content under ambient tropical condition

Maize (cv. TSZB) samples were re-moistened to different moisture contents (m.c.s) of 13, 15, 17, 20, 25, 30 or 35% and stored with the natural microflora or sterilized before artificial inoculation with either single or mixed moulds (Aspergillus flavus, A. niger, Penicilium purpurogenum and Fusarium moniliforme) and evaluated for initiation time for moulding, fungal populations and aflatoxin B₁ production. Whereas the fungal populations of naturally contaminated maize of 13% m.c. decreased significantly with storage, 17 and 20% m.c. maize increased with the latter showing maximum of about log₁₀ 7 colony forming units (cfu g⁻¹). Of the samples (13, 15, 17 or 20% m.c. maize), only those of ≥20% m.c. showed hazardous levels (>20 ppb) of aflatoxin B₁ production. The 20% m.c. sample also showed a significant positive correlation (r = 0.92) between m.c. and fungal load but those of lower m.c.s exhibited poor correlations, probably reflecting the absence of changes in the m.c.s of the 13, 15 and 17% m.c. maize. Aflatoxin B₁ content of 25% m.c. maize increased with increase in inoculum concentration of A. flavus. Mixed mould inoculation of maize samples resulted in a reduction in aflatoxin concentration with co-cultures of A. flavus and P. purpurogenum showing the lowest production, while that inoculated with A. flavus alone (control) exhibiting the maximum production. Initiation time for moulding was most rapid in ≥20% m.c. maize irrespective of inoculum type, with A. flavus being the most invasive in singly inoculated samples. However, A flavus was most competitive in 20–30% m.c. maize inoculated with mixed moulds, while F. moniliformewas most competitive in the 35% m.c. maize.     

Are There any Risks in Using Entomopathogenic Fungi for Pest Control,with Particular Reference to the Bioactive Metabolites of Metarhizium,Tolypocladium and Beauveria species?

Entomopathogenic fungi are promising alternatives to chemical insecticides. However, a major hurdle concerning the registration of these fungi as plant protection agents is the possible toxicity of secreted metabolites, especially secondary metabolites. This review summarizes data on specific secondary metabolites (destruxins, efrapeptins, oosporein, beauvericin and beauveriolides) produced by the important genera Beauveria, Metarhizium and Tolypocladium . The quantities of secondary metabolites produced by these fungi in vivo are usually much less than those secreted in nutrient rich liquid media. Methods and strategies are suggested which could standardize the risk assessment of fungal biological control agents.     

Novel Fusarium head blight pathogens from Nepal and Louisiana revealed by multilocus genealogical concordance

This study was conducted to assess evolutionary relationships, species diversity and trichothecene toxin potential of five Fusarium graminearum complex (FGSC) isolates identified as genetically novel during prior Fusarium head blight (FHB) surveys in Nepal and Louisiana. Results of a multilocus genotyping (MLGT) assay for B-trichothecene species determination indicated these isolates might represent novel species within the FGSC. GCPSR-based phylogenetic analyses of a 12-gene dataset, comprising portions of seven loci totaling 13.1 kb of aligned DNA sequence data, provided strong support for the genealogical exclusivity of the Nepalese and Louisianan isolates. Accordingly, both species are formally recognized herein as novel FGSC species. Fusarium nepalense was resolved as the sister lineage of Fusarium ussurianum + Fusarium asiaticum within an Asian subclade of the FGSC. Fusarium louisianense was strongly supported as a reciprocally monophyletic sister of Fusarium gerlachii + F. graminearum, suggesting that this subclade might be endemic to North America. Multilocus Bayesian species tree analyses augment these results and provide evidence for a distinct lineage within F. graminearum predominately from the Gulf Coast of Louisiana. As predicted by the MLGT assay, mycotoxin analyses demonstrated that F. nepalense and F. louisianense could produce 15ADON and nivalenol, respectively, in planta. In addition, both species were only able to induce mild FHB symptoms on wheat in pathogenicity experiments.     

Production of cyclopiazonic acid,aflatrem, and aflatoxin by <Emphasis Type="Italic">Aspergillus flavus</Emphasis> is regulated by <Emphasis Type="Italic">veA</Emphasis>, a gene necessary for sclerotial formation

The plant pathogenic fungus Aspergillus flavus produces several types of mycotoxins. The most well known are the carcinogenic compounds called aflatoxins. In addition, A. flavus produces cyclopiazonic acid and aflatrem mycotoxins, contributing to the toxicity of A. flavus infected crops. Cyclopiazonic acid is a specific inhibitor of calcium-dependent ATPase in the sarcoplasmic reticulum that results in altered cellular Ca⁺⁺ levels. Aflatrem is a potent tremorgenic mycotoxin known to lead to neurological disorders. Previously we showed that a gene called veA controls aflatoxin and sclerotial production in A. parasiticus. In this study in A. flavus, we show that the veA homolog in A. flavus not only is necessary for the production of aflatoxins B1 and B2 and sclerotia, but also regulates the synthesis of the mycotoxins cyclopiazonic acid and aflatrem. The A. flavus ΔveA mutant was completely blocked in the production of aflatrem and showed greater than twofold decrease in cyclopiazonic acid production. The genes involved in the synthesis of cyclopiazonic acid are unknown; however, the aflatrem gene cluster has been characterized. Northern hybridization analysis showed that veA is required for expression of the A. flavus aflatrem genes atmC, atmG, and atmM. This is the first report of a regulatory gene governing the production of cyclopiazonic acid and aflatrem mycotoxins.     

Evaluation of biological control formulations to reduce aflatoxin contamination in peanuts

A two-year study was conducted to evaluate the efficacy of three formulations of nontoxigenic strains of Aspergillus flavus and Aspergillus parasiticus to reduce preharvest aflatoxin contamination of peanuts. Formulations included: (1) solid-state fermented rice; (2) fungal conidia encapsulated in an extrusion product termed Pesta; (3) conidia encapsulated in pregelatinized corn flour granules. Formulations were applied to peanut plots in 1996 and reapplied to the same plots in 1997 in a randomized design with four replications, including untreated controls. Analysis of soils for A. flavus and A. parasiticus showed that a large soil population of the nontoxigenic strains resulted from all formulations. In the first year, the percentage of kernels infected by wild-type A. flavus and A. parasiticus was significantly reduced in plots treated with rice and corn flour granules, but it was reduced only in the rice-treated plots in year two. There were no significant differences in total infection of kernels by all strains of A. flavus and A. parasiticus in either year. Aflatoxin concentrations in peanuts were significantly reduced in year two by all formulation treatments with an average reduction of 92%. Reductions were also noted for all formulation treatments in year one (average 86%), but they were not statistically significant because of wide variation in the aflatoxin concentrations in the untreated controls. Each of the formulations tested, therefore, was effective in delivering competitive levels of nontoxigenic strains of A. flavus and A. parasiticus to soil and in reducing subsequent aflatoxin contamination of peanuts.     

Vorkommen von ochratoxin A und B in gewürzen

A total of 681 samples of spices, which comprised more than 50 different spice commodities were analysed for the natural occurrence of the mycotoxins ochratoxin A (OTA) and ochratoxin B (OTB). The analytical method involved chloroform extraction, clean-up by immunoaffinity column and HPLC determination of both mycotoxins. OTA and OTB were detected in 143 (21%) and 68 (10%) of the samples, respectively. The highest frequency of occurrence of both mycotoxins detected were in chili (100% for OTA and 55% for OTB), paprika (41% and 15%, respectively) and pepper (23% and 44%, respectively). The toxin concentrations ranged between the detection limit (0.01 ng/g) and 41.8 ng OTA (2.7 ng OTB)/g of chili, 18.9 ng OTA (1.4 ng OTB)/g of paprika and 3.8 ng OTA (4.6 ng OTB)/g of pepper. One sample of a extract of vanilla was found to be positive for OTB at 15 ng/g. However, median values of most samples showed to be below the detection limit. Comparison of the geographical origin of the samples showed that the predominant number of contaminated spices was from Southeast-Asia and India. Highly contaminated paprika samples were found to come from Israel.     

大豆连作障碍研究Ⅰ．大豆连作土壤紫青霉菌的毒素作用研究

研究分析了大豆连作、轮作土壤微生物区系,发现连作大豆根际土壤真菌富集,以其优势真菌回接大豆．紫青霉菌（Ｐｅｎｉｃｉｌｌｉｕｍｐｕｒｐｕｒｏｇｅｎｕｍ）能强烈抑制大豆生长发育．在实验室条件下分离获得该菌产生的毒素粗结晶,５μｇ·ｍｌ－１水培液中即可观察到大豆根系受害,根毛很少生长;３０μｇ·ｍｌ－１水培液中大豆主根褐变严重,侧根几乎不再生长;２００μｇ·ｍｌ－１导致一些大豆品种幼苗在２周内死亡这些结果表明,连作大豆土壤中该菌的大量存在及其产生的毒素是大豆连作障碍产生的主要因素．     

丝状真菌G蛋白信号途径的研究进展

丝状真菌在工业、农业、医药等领域具有重要经济价值,一些亦可导致人类及动、植物疾病,造成经济损失.G蛋白信号途径是真核生物中普遍存在的细胞跨膜信号转导途径.近年来,丝状真菌中G蛋白信号途径的研究发展很快,相关报道表明该信号途径参与感应并传递多种胞外信号刺激,对丝状真菌的生长、分化、繁殖、致病性及真菌毒素等次生代谢产物合成有重要的调控作用.本文就丝状真菌中G蛋白信号途径的基本组成及其生理功能的研究现状进行简要综述.     

In vitro vomitoxin exposure alters IgA and IgM secretion by CH12LX B cells

The CH12LX cell line was used as a clonal model to assess the direct effects of vomitoxin on IgM and IgA secretion in B cells. When vomitoxin was included in LPS-driven CH12LX B cell cultures, it had multiple effects on Ig secretion. Whereas vomitoxin doses of 115 and 120 ng/ml caused 50% inhibition(ID₅₀) of IgA and IgM production, respectively, toxin concentrations in the 5 to 50 ng/ml range slightly stimulated IgA production. However, low vomitoxin doses did not induce switching of membrane IgM+ CH12LX B cells to membrane IgA+. Total cell number was unaffected at vomitoxin concentrations up to 100 ng/ml but dropped markedly at 200 ng/ml (ID₅₀=170 ng/ml). Using the MTT reduction assay as another measure of viability and cell function, vomitoxin was also inhibitory (ID₅₀=130 ng/ml). Both thymidine incorporation and leucine incorporation were also inhibited by the toxin with estimated ID₅₀s being 120 and 110 ng/ml, respectively. The results indicate that although at high doses, vomitoxin inhibits proliferation, Ig secretion and DNA/protein synthesis in the clonal B cell model, the toxin marginally stimulated IgA secretion at lower doses.     

The concentration and prevalence of ochratoxin A in coffee and coffee-based products: A global systematic review,meta-analysis and meta-regression

The current investigation was aimed to estimate the prevalence and concentration of ochratoxin A (OTA) in different types of coffee and coffee-based products with the aid of a systematic review and meta-analysis. Therefore, the recommended databases including PubMed, Scopus, and Embase from Jan 1983 to Oct 2018 were screened to retrieve the related citations. In this regard, among 1041 explored articles in the identification step, thirty six articles with 3182 samples were included in the meta-analysis and meta-regression. According to findings, the global pooled concentration and prevalence of OTA was calculated as 3.21 μg/kg (95% CI: 3.08–3.34 μg/kg) and 53.0 % (95% CI: 43.0–62.0), respectively. Also, direct correlations between the increases in poverty as well as the amount of annual precipitation and prevalence of OTA was noted, while with decreasing in HDI the prevalence of OTA in coffee significantly was increased. Moreover, the lowest and highest concentrations of OTA in coffee were observed in Taiwan (0.35 μg/kg) and Turkey (79.0 μg/kg), respectively. The outcome of this meta-analysis can be used for the building of risk assessment models aiming to derive data for the development of specific actions to reduce the exposure to this mycotoxin in coffee and coffee-based products.