Management and prevention of mycotoxins in peanuts

Contamination of peanuts with mycotoxins, particularly aflatoxins, is a worldwide problem that affects both food safety and agricultural economies. Most countries have adopted regulations that limit the quantity of aflatoxins in food and feed to 20 µg kg^-1 or less; however, environmental conditions in most of the world where peanuts are produced and stored often make it difficult or impossible to attain such low concentrations. In addition to aflatoxins, peanuts are often contaminated with cyclopiazonic acid (CPA). Both mycotoxins are produced by Aspergillus flavus, a ubiquitous fungus that can infect and grow in peanuts under both pre- and post-harvest conditions. Management of mycotoxin contamination in peanuts generally involves removal of high-risk components from shelled lots or the removal of individual, highly contaminated nuts. This is accomplished by various processes such as screening, kernel sizing, electronic colour sorting, hand sorting, and blanching followed by electronic colour sorting. Recently, biological control technology has been developed that prevents much of the contamination that might otherwise occur. Biocontrol is based on competitive exclusion whereby a dominant population of a non-toxigenic strain of A. flavus is established in the soil before peanuts are subjected to conditions favouring contamination. The applied strain competes with toxigenic strains for infection sites, resulting in significantly reduced concentrations of aflatoxins in peanuts. Monitoring of the first commercial use of the technology showed that aflatoxins were reduced by an average of 85% in farmers' stock peanuts and by as much as 98% in shelled, edible grade peanuts.     

QuEChERS结合超高效液相色谱-串联质谱法测定砂仁中4种黄曲霉毒素

该研究建立了利用QuEChERS-超高效液相色谱串联质谱（UPLC-MS/MS）法同时测定砂仁中4种黄曲霉毒素（Aflatoxin B1、B2、G1、G2）的分析方法。通过对QuEChERS前处理技术、色谱和质谱条件的优化，确定了最佳的实验条件。研磨后的样品经φ=1%甲酸酸化的乙腈提取，无水硫酸镁及氯化钠脱水盐析，上清液经十八烷基键合硅胶（C18）、乙二胺-Ｎ-丙基硅烷（PSA）、无水硫酸镁混合吸附剂净化。经ACQUITY UPLC HSS T3色谱柱（1.8 µm，2.1 mm×100 mm）进行分离，以乙腈和φ=0.1%甲酸-5 mmol/L乙酸铵溶液进行梯度洗脱。在正离子模式下，进行多反应监测（MRM），进一步通过空白基质标准溶液外标法定量。结果表明，4种黄曲霉毒素在0.20~10.00 μg/L范围内线性关系良好，相关系数均大于0.998 9。实际样品的加标回收试验结果显示，回收率范围为89.50%~113.12%，日内精密度为1.31%~6.71%，日间精密度为1.29%~6.20%，4种黄曲霉毒素的方法检出限为0.30~0.60 μg/kg、定量限为1.00~2.00 μg/kg。该方法操作简单，快速，灵敏，适用于砂仁中4种黄曲霉毒素的定性、定量筛查。     

组合实验设计用于黄曲霉毒素污染控制的研究

为控制黄曲霉毒素主要产生菌寄生曲霉 AS3.4407在粮食储藏环境中的产毒污染,采用组合实验设计策略,考察了五个模拟环境因子对菌株产毒的影响。首先,利用 Plack-ett-Burman 试验筛选出三个对毒素产量影响显著的因子(转速、温度、pH),在此基础上,采用CCD 实验设计法(Central Composite Design,CCD)研究了这三个因子对寄生曲霉 AS3.4407产毒影响的交互作用,并根据毒素曲面方程和二次多项回归方程得到该菌的最低产毒条件。在环境通风良好(转速240rpm),储藏温度较低(18℃),pH 为弱碱性(pH8)条件下,寄生曲霉AS3.4407产毒量最低,验证试验证实了该方程的预测值与实际值之间具有较好的拟合度。为粮库采取相应措施,改变储藏环境条件以降低毒素污染提供了很好依据。     

酱油中黄曲霉毒素B1检测方法的探讨

利用免疫亲和柱净化维康4系列荧光计检测酱油中黄曲霉毒素B1，添加回收率在90％以上。该方法操作简单安全，定量快速准确。     

黄曲霉毒素B1降解菌的筛选鉴定及发酵条件优化

该研究采用香豆素筛选模型,从酒醅、大曲、榨菜、酸豆角等样品中分离筛选降解黄曲霉素B1（AFB1）菌株,通过形态学观察及分子生物学技术对筛选菌株进行鉴定,并以黄曲霉毒素B1降解率为评价指标,采用单因素及正交试验对筛选菌株的发酵条件进行优化。结果表明,共初筛出23株可利用香豆素菌株;采用高效液相色谱（HPLC）法复筛出1株AFB1高效降解菌,其降解率最高,为86.22%,菌株编号为HB022。菌株HB022被鉴定为地衣芽孢杆菌（Bacillus licheniformis）,其最优发酵条件为发酵温度37 ℃、初始pH 7.2、接种量5%、发酵时间72 h。在此优化条件下,AFB1的降解率为91.13%。     

Feasibility of Detecting Aflatoxin B1 on Inoculated Maize Kernels Surface using Vis/NIR Hyperspectral Imaging

The feasibility of using a visible/near‐infrared hyperspectral imaging system with a wavelength range between 400 and 1000 nm to detect and differentiate different levels of aflatoxin B₁ (AFB₁) artificially titrated on maize kernel surface was examined. To reduce the color effects of maize kernels, image analysis was limited to a subset of original spectra (600 to 1000 nm). Residual staining from the AFB₁ on the kernels surface was selected as regions of interest for analysis. Principal components analysis (PCA) was applied to reduce the dimensionality of hyperspectral image data, and then a stepwise factorial discriminant analysis (FDA) was performed on latent PCA variables. The results indicated that discriminant factors F₂ can be used to separate control samples from all of the other groups of kernels with AFB₁ inoculated, whereas the discriminant factors F₁ can be used to identify maize kernels with levels of AFB₁ as low as 10 ppb. An overall classification accuracy of 98% was achieved. Finally, the peaks of β coefficients of the discrimination factors F₁ and F₂ were analyzed and several key wavelengths identified for differentiating maize kernels with and without AFB₁, as well as those with differing levels of AFB₁ inoculation. Results indicated that Vis/NIR hyperspectral imaging technology combined with the PCA–FDA was a practical method to detect and differentiate different levels of AFB₁ artificially inoculated on the maize kernels surface. However, indicated the potential to detect and differentiate naturally occurring toxins in maize kernel.     

不同前处理方法对花生粕酶解液中黄曲霉毒素含量的影响

研究了弱碱高温处理法、H_2O_2处理法和不同过滤介质去除黄曲霉毒素的效果。结果表明,弱碱高温处理对原料中的黄曲霉毒素有较好的去除效果,花生粕经过121℃,pH10,60min处理,黄曲霉毒素的破坏率高达84.5%,最终酶解上清液中黄曲霉毒素的含量为0.344ng/mL,达到欧盟安全标准;用H_2O_2处理,低浓度H_2O_2对于黄曲霉毒素的破坏很少,当浓度达到1600mg/kg,酶解上清液中的黄曲霉毒素达到0.104ng/mL,相对于不加H_2O_2处理的酶解上清液,其黄曲霉毒素减少率达94.87%,但高浓度的H_2O_2处理不利干酶解的进行;对于不同的过滤介质,以活性碳吸附去毒效果较好,但同时氨态氮的损失率高达7.03%。综合考虑实验的各方面因素,如成本问题以及脱毒后酶解的蛋白回收率和氨态氮含量问题,选择先在121℃,pH10条件下处理60min,酶解48h,然后用普通滤纸过滤进行脱毒处理,此时酶解上清液中的黄曲霉毒素达到0.244ng/mL。     

Aflatoxin B1 and aflatoxins in ground red chilli pepper after drying

In this study, 180 red chilli pepper (RCP) berry samples were obtained from two different croplands of Gaziantep and Kahramanmara? (Turkey) in August, September and October. RCP berry samples were dried under sunlight and grinded. Ground red chilli pepper (GRCP) samples were analysed for aflatoxins (AFs, sum of B₁, B₂, G₁ and G₂) and AFB₁ contamination. According to the results, in 49 of 180 samples, AFB₁ and in 37 samples, AFs were higher than legal limits. The lowest amounts of AFs and AFB₁ were obtained in August and the highest amounts in October. χ² analysis showed that there were no significant differences (p > 0.05) between cities among 3 months according to number of samples with AFs and AFB₁ above legal limits. According to the Duncan multiple-range test, there was no significant difference between all months. Strict measures are necessary to produce high-quality GRCP. RCP berry must be treated to reduce moulds before production of GRCP.     

Pediococcus acidolactici and Pediococcus pentosaceus isolated from a rainbow trout ecosystem have probiotic and ABF1 adsorbing/degrading abilities in vitro

ABSTRACT

Probiotics are being used in biological control of bacterial pathogens, as an alternative to antibiotics, to improve health and production parameters in fish farming. Fish farming production is severely affected by aflatoxins (AFs), which are a significant problem in aquaculture systems. Aflatoxins exert substantial impact on production, causing disease with high mortality and a gradual decline of reared fish stock quality. Some aspects of aflatoxicosis in fish, particularly its effects on the gastrointestinal tract, have not been well documented. The aim of the present study was to evaluate probiotic properties of lactic acid bacterial (LAB) strains isolated from rainbow trout intestine and feed. Moreover, AFB₁-binding and/or degrading abilities were also evaluated to assess their use in the formulation of feed additives. Growth at pH 2, the ability to co-aggregate with bacterial pathogens, inhibition of bacterial pathogens, and determination of the inhibitory mechanism were tested. Aflatoxin B₁ (AFB₁) adsorption and degradation ability were also tested. All strains were able to maintain viable (10⁷ cells ml^–1) at pH 2. Pediococcus acidilactici RC001 and RC008 showed the strongest antimicrobial activity, inhibiting all the pathogens tested. The strains produced antimicrobial compounds of different nature, being affected by different treatments (catalase, NaOH and heating), which indicated that they could be H₂O_2, organic acids or proteins. All LAB strains tested showed the ability to coaggregate pathogenic bacteria, showing inhibition percentages above 40%. Pediococcus acidilactici RC003 was the one with the highest adsorption capacity and all LAB strains were able to degrade AFB₁ with percentages higher than 15%, showing significant differences with respect to the control. The ability of some of the LAB strains isolated in the present work to compete with pathogens, together with stability against bile and gastric pH, reduction of bioavailability and degradation of AFB₁, may indicate the potential of LAB for use in rainbow trout culture.