检测角度对激光诱导荧光光谱技术检测花生油AFB1污染的影响研究

近年来,近红外光谱、高光谱等快速无损检测技术,被广泛应用于农产品中黄曲霉毒素B₁(Aflatoxin B₁, AFB₁)的检测。但这些方法主要是根据黄曲霉改变样品本身的结构结合化学计量学方法间接检测样品中AFB₁,无特异性。因此,本研究拟利用研发的激光诱导荧光(laser induce fluorescence, LIF)技术获取污染AFB₁花生油的荧光光谱,建立精确无损检测花生油中AFB₁的定性、定量方法。首先,人工制备7种不同AFB₁污染水平的花生油样品,同时设置对照组。其次利用研发的LIF系统从不同角度检测样品的荧光光谱信号。最后,对获取的光谱进行分析。结果显示,三个检测角度的荧光强度在400-800 nm内均随着AFB₁污染水平的增加而增加,定性和定量的结果均是当检测荧光角度为90度时表现最优。表明当检测角度为90度时,LIF技术可以作为一种快速、精确的方法来判定花生油中AFB₁污染水平。     

Distribution of aflatoxins and fumonisins in dry-milled maize fractions

The aim was to evaluate the distribution of aflatoxins and fumonisins in fractions derived from the dry-milling of contaminated maize. Two maize lots with different contamination levels were processed and sampled: the first (maize 1) had aflatoxin B₁ (AFB₁) and fumonisin B₁ (FB₁) levels of 3.6 and 5379 µg kg^?1, respectively; the second (maize 2) had corresponding levels of 91.1 and 8841 µg kg^?1, respectively. The cleaning step reduced AFB₁ and FB₁ levels by 8 and 11% in maize 1 and by 57 and 34% in maize 2. The subsequent removal of bran and germ led to a further decrease in contamination levels in the products destined for human consumption. In the latter, AFB₁ was uniformly distributed, while FB₁ was concentrated in the finer size fractions. Contamination of raw maize 1 (3.6 µg kg^?1) was below the European Union AFB₁ limit of 5 µg kg^?1 for unprocessed maize, but among the final products only coarse flour (1.7 µg kg^?1) was within the European Union limit of 2 µg kg^?1, while grits and fine flour showed higher levels (2.7 and 2.5 µg kg^?1, respectively). As regards cleaned maize, a different distribution of the two toxins was observed in the kernels: AFB₁ contamination was more superficial and concentrated in germ, while FB₁ contamination affected the inner layers of the kernels.     

基于高光谱图像光谱变量和颜色特征的霉变玉米籽粒识别

目的:准确识别霉变玉米籽粒。方法:基于高光谱图像光谱变量和颜色特征建立霉变玉米籽粒识别的新方法。先对玉米籽粒图像进行图像分割和光谱变量、颜色特征提取,并根据颜色特征生成颜色直方图;将光谱变量和颜色直方图特征组成特征集合;通过距离函数对特征集合中所有特征的分析确定霉变玉米籽粒所属类别。结果:所提方法对霉变玉米籽粒类别的最大平均识别偏差为1.12,最佳平均识别准确率为97.59%;与基于高光谱图像+随机蛙跳+极限学习机的方法、基于高光谱图像+稀疏自动编码器+卷积神经网络的方法、基于高光谱图像+蚁群优化+BP神经网络的方法相比,研究所提方法对霉变玉米籽粒类别的识别准确率明显提高。结论:该方法可实现被测玉米籽粒样品是否霉变以及霉变程度的准确判断。     

Detection of fungal infection and Ochratoxin A contamination in stored wheat using near-infrared hyperspectral imaging

A study was done to detect Aspergillus glaucus, and Penicillium spp., infection and Ochratoxin A contamination in stored wheat using a Near-Infrared (NIR) Hyperspectral Imaging system. Fungal-infected samples were imaged every two weeks, and the three dimensional hypercubes obtained from image data were transformed into two dimensional data. Principal component analysis was applied to the two dimensional data and based on the highest factor loadings, 1280, 1300, and 1350 nm were identified as significant wavelengths. Six statistical features and ten histogram features corresponding to the significant wavelengths were extracted and subjected to linear, quadratic and Mahalanobis discriminant classifiers. All the three classifiers differentiated healthy kernels from fungal-infected kernels with a classification accuracy of more than 90%. The quadratic discriminant classifier provided classification accuracy higher than the linear and Mahalanobis classifiers for pair-wise, two-way and six-way classification models. The Ochratoxin A contaminated samples had a unique significant wavelength at 1480 nm in addition to the two significant wavelengths corresponding to fungal infection. The peak at 1480 nm was identified only in the Ochratoxin A contaminated samples. The Ochratoxin A contaminated samples can be detected with 100% classification accuracy using NIR hyperspectral imaging system. The NIR hyperspectral system can differentiate between different fungal infection stages and different levels of Ochratoxin A contamination in stored wheat.     

基于监测过氧化氢酶活性对储藏玉米AFB1污染的早期预警

早期预警是防控储藏玉米污染黄曲霉毒素B1(AFB_1)的重要手段。将不同含水量的玉米置于各种温度环境下进行储藏试验,研究过氧化氢酶活性变化与玉米污染黄曲霉毒素B1(AFB_1)的关系,结果表明,两者数据变化的趋势一致,AFB_1是过氧化氢酶活性的二次曲线函数(相关性系数大于0.95)。储藏温度和玉米含水量等条件可改变所含霉菌产生AFB_1和升高过氧化氢酶活性的速度,但不影响数值变化的趋势。在各种储藏条件下,过氧化氢酶活性变化可比玉米中AFB_1含量显著变化的起始点提前4~21d。因此,借助检测过氧化氢酶活性的方法可以早期预警储藏玉米污染AFB_1。     

Correlation and classification of single kernel fluorescence hyperspectral data with aflatoxin concentration in corn kernels inoculated with Aspergillus flavus spores

The objective of this study was to examine the relationship between fluorescence emissions of corn kernels inoculated with Aspergillus flavus and aflatoxin contamination levels within the kernels. Aflatoxin contamination in corn has been a long-standing problem plaguing the grain industry with potentially devastating consequences to corn growers. In this study, aflatoxin-contaminated corn kernels were produced through artificial inoculation of corn ears in the field with toxigenic A. flavus spores. The kernel fluorescence emission data were taken with a fluorescence hyperspectral imaging system when corn kernels were excited with ultraviolet light. Raw fluorescence image data were preprocessed and regions of interest in each image were created for all kernels. The regions of interest were used to extract spectral signatures and statistical information. The aflatoxin contamination level of single corn kernels was then chemically measured using affinity column chromatography. A fluorescence peak shift phenomenon was noted among different groups of kernels with different aflatoxin contamination levels. The fluorescence peak shift was found to move more toward the longer wavelength in the blue region for the highly contaminated kernels and toward the shorter wavelengths for the clean kernels. Highly contaminated kernels were also found to have a lower fluorescence peak magnitude compared with the less contaminated kernels. It was also noted that a general negative correlation exists between measured aflatoxin and the fluorescence image bands in the blue and green regions. The correlation coefficients of determination, r ², was 0.72 for the multiple linear regression model. The multivariate analysis of variance found that the fluorescence means of four aflatoxin groups, <1, 1–20, 20–100, and ≥100 ng g^?1 (parts per billion), were significantly different from each other at the 0.01 level of alpha. Classification accuracy under a two-class schema ranged from 0.84 to 0.91 when a threshold of either 20 or 100 ng g^?1 was used. Overall, the results indicate that fluorescence hyperspectral imaging may be applicable in estimating aflatoxin content in individual corn kernels.     

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.     

Aflatoxin accumulation in whole crop maize silage as a result of aerobic exposure

BACKGROUND: Most of the maize silage stored in horizontal silos is exposed to air and can be spoiled by fungi. Potentially toxigenic fungi have been found in maize silage, and about 300 mycotoxins have been detected. Among these mycotoxins, the most harmful for feed and food safety are aflatoxins. The aim of the study was to set up a specific method to detect aflatoxins in maize silage, and to investigate whether aflatoxin contamination in maize silage depends on the level of field contamination of the crop, and whether the occurrence of aerobic spoilage during ensiling has any effect on the final contamination of the silage. RESULTS: A method for the determination of aflatoxin B₁, B₂, G₁ and G₂ in maize silage using high‐performance liquid chromagraphy with fluorescence detection has been developed and validated. Recoveries of aflatoxin B₁, B₂, G₁, and G₂ spiked over the 0.25 to 5 µg kg^?1 range averaged 74–94%. The results of laboratory scale and farm scale ensiling experiments indicated that aflatoxins could increase when silage is exposed to air during conservation or during the feed‐out phase. CONCLUSIONS: The method here proposed to detect aflatoxins in silages has proved to be sensitive and is able to detect levels of 0.1 and 0.5 ng mL^?1 for AFB₁ and AFG₁, and between 0.025 and 0.125 ng mL^?1 for AFB₂ and AFG₂. This study also provides evidence of aflatoxin accumulation in whole crop maize silage as a result of aerobic exposure. Copyright © 2011 Society of Chemical Industry     

Transfer of aflatoxin B1 and fumonisin B1 from naturally contaminated raw materials to beer during an industrial brewing process

The aim of this research was to determine the fate of aflatoxins (AFs) and fumonisins (FBs) naturally occurring in raw materials (maize grit and malted barley) during four industrial brewing processes. The aflatoxin B₁ (AFB₁) level in raw materials varied from 0.31 to 14.85 µg kg^?1, while the fumonisin B₁ (FB₁) level (only in maize grit) varied from 1146 to 3194 µg kg^?1. The concentration in finished beer ranged from 0.0015 to 0.022 µg l^?1 for AFB₁ and from 37 to 89 µg l^?1 for FB₁; the other aflatoxins and fumonisin B₂ were not found in beer samples. The average percentage of toxins recovered in finished beer, referring to the amounts contained in raw materials, were 1.5% ± 0.8% for AFB₁ and 50.7% ± 4.7% for FB₁. These results were mainly due to the different solubility of the two mycotoxins during the mashing process. If raw materials comply with the limits fixed by European Commission Regulations, the contribution of a moderate daily consumption of beer to AFB₁ and FB₁ intake does not contribute significantly to the exposure of the consumer.     

Aflatoxin in household maize for human consumption in Kenya,East Africa

ABSTRACT

The objective of this study is to determine the occurrence and level of aflatoxins (AFs) contamination in freshly harvested maize for human consumption in rural Kenya. Maize kernels and freshly milled maize flour (n = 338) were collected from households in Siaya and Makueni counties. While both counties are representatives of different environmental and climate conditions, Makueni County is the area with reported outbreaks of aflatoxicosis. Samples were analysed for AFB₁, AFB₂, AFG₁, and AFG₂ using Ultra High-Pressure Liquid Chromatography with Fluorescence detection. AFs were detected in 100% of the samples with the range of 2.14–411 µg/kg. The geometric mean of total AFs in all samples from Makueni County is 62.5 μg/kg with 95% CI: 53.7, 71.4 while in Siaya County is 52.8 μg/kg with 95% CI: 44.0, 61.7. This study showed that AFs contamination is prevalent in maize-based foods in the region.     

Mycotoxin contamination of home-grown maize in rural northern South Africa (Limpopo and Mpumalanga Provinces)

The aim of this study was to assess mycotoxin contamination of crops grown by rural subsistence farmers over two seasons (2011 and 2012) in two districts, Vhembe District Municipality (VDM, Limpopo Province) and Gert Sibande District Municiality (GSDM, Mpumalanga Province), in northern South Africa and to evaluate its impact on farmers’ productivity and human and animal health. A total of 114 maize samples were collected from 39 households over the two seasons and were analysed using a validated liquid chromatography-tandem mass spectrometry mycotoxins method. Aflatoxin B₁ (AFB₁) occurrence ranged from 1 to 133 µg kg^?1 in VDM while AFB₁ levels in GSDM were less than 1.0 µg kg^?1 in all maize samples. Fumonisin B₁ levels ranged from 12 to 8514 µg kg^?1 (VDM) and 11–18924 µg kg^?1 (GSDM) in 92% and 47% positive samples, respectively, over both seasons. Natural occurrence and contamination with both fumonisins and aflatoxins in stored home-grown maize from VDM was significantly (p < 0.0001) higher than from GSDM over both seasons.     

The detection of aflatoxin B1 in peanut kernels,peanut butter and maize using a monoclonal antibody based enzyme immunoassay

A simple procedure was validated for the routine immunochemical analysis of AFB₁ in peanut kernels, peanut butter and maize. The specificity, affinity and sensitivity of the monoclonal antibody (McAb) employed was such that minimal sample preparation was required. The enzyme immunoassay (EIA) had a sensitivity for standard AFB₁ of 0·2 ng ml⁻¹ with a working range up to 30 ng ml⁻¹ and was not significantly affected by matrix interference of samples. Essential protocol features were (1) blending substrate with methanol: water; (2) filtering blended sample; and (3) analysis of filtrate by EIA after dilution with buffer. Average recoveries of AFB₁ spiked samples at levels of 6–400 ppb were 90–112·5%. Using laboratory prepared samples contaminated with Aspergillus flavus there was high positive correlation (r = 0·97) when EIA results were compared with thin layer chromatography (TLC) techniques.     

Fungal contamination and aflatoxin content of maize,moringa and peanut foods from rural subsistence farms in South Haiti

Mycotoxins are toxic, low molecular weight compounds produced by fungi. Among them, aflatoxins are the most carcinogenic and they mainly impact on rural communities of developing countries. The present study supplies data on mycobiota and aflatoxin contamination in the most common food products consumed in Haiti. The study concerns analyses performed on 49 samples of meals and seeds collected in South Haiti and tested for fungal occurrence and aflatoxin content by HPLC-DAD technique. The results revealed that three main fungal genera affected Haitian food products: Aspergillus spp. (Section Flavi and Nigri), followed by Penicillium spp. and Fusarium spp. Aflatoxin was present in more than half of the samples of maize (Zea mays L.) kernels (55%), maize meal (57%) and moringa (Moringa oleifera Lam.) seeds (64%), and in 25% of peanut (Arachis hypogaea L.) samples. The tested food products were mostly contaminated by aflatoxin B₁ (AFB₁) followed by aflatoxin B₂ (AFB₂), while no aflatoxins type G were detected. The total concentration of aflatoxins in the positive samples was 228 μg/kg on average, i.e., fifty-seven and eleven times higher than the maximum levels allowed in Europe and USA, respectively. Both the presence of aflatoxigenic fungi and aflatoxin contamination in maize kernels seemed to be related to agricultural practices, such as weed control, irrigation and growing cycle length. These findings suggest that the Haitian population is strongly exposed to aflatoxin risk. This risk could be reduced by exploiting simple and accessible farming strategies for minimizing mycotoxin contamination, at least for maize.     

Application of Image Texture Analysis for Evaluation of X-Ray Images of Fungal-Infected Maize Kernels

The feasibility of image texture analysis to evaluate X-ray images of fungal-infected maize kernels was investigated. X-ray images of maize kernels infected with Fusarium verticillioides and control kernels were acquired using high-resolution X-ray micro-computed tomography. After image acquisition and pre-processing, several algorithms were developed to extract image textural features from selected two-dimensional (2D) images of the kernels. Four first-order statistics (mean, standard deviation, kurtosis and skewness) and four grey level co-occurrence matrix (GLCM) features (correlation, energy, homogeneity and contrast) were extracted from the side, front and top views of each kernel and used as inputs for principal component analysis (PCA). The first-order statistical image features gave a better separation of the control from infected kernels on day 8 post-inoculation. Classification models were developed using partial least squares discriminant analysis (PLS-DA), and accuracies of 67 and 73% were achieved using first-order statistical features and GLCM extracted features, respectively. This work provides information on the possible application of image texture as method for analysing X-ray images.     

Effect of different ozone treatments on aflatoxin degradation and physicochemical properties of pistachios

This study evaluated the efficiency of ozone for the degradation of aflatoxins in pistachio kernels and ground pistachios. Pistachios were contaminated with known concentrations of aflatoxin (AF) B₁, B₂, G₁ and G₂. Pistachio samples were exposed to gaseous ozone in a chamber at 5.0, 7.0 and 9.0 mg L^?1 ozone concentrations for 140 and 420 min at 20 °C and 70% RH. Aflatoxin degradation was determined by high performance liquid chromatography (HPLC). The efficiency of ozone for aflatoxin degradation in pistachios increased with increasing exposure time and ozone concentration. The results indicated that AFB₁ and total aflatoxins could be reduced by 23 and 24%, respectively, when pistachio kernels were ozonated at 9.0 mg L^?1 ozone concentration for 420 min. Only a 5% reduction in AFB₁ and total aflatoxin levels could be achieved for ground pistachios under the same conditions. No significant changes occurred in pH, color, moisture content and free fatty acid values of pistachio kernels and ground pistachios. Fatty acid compositions of pistachios did not change significantly after the ozonation treatments. No significant changes were found between sweetness, rancidity, flavor, appearance and overall palatability of ozonated and non‐ozonated pistachio kernels. Significant changes were observed in the organoleptic properties of ground pistachios, except rancidity, after 5.0 mg L^?1 ozone treatment for 140 min. Ozonation was found to be more effective for degrading aflatoxins in pistachio kernels than ground pistachios. Copyright © 2006 Society of Chemical Industry     

Detection of fusarium damage in Canadian wheat using visible/near-infrared hyperspectral imaging

Fusarium damage in wheat may reduce the quality and safety of food and feed products. In this study, the use of hyperspectral imaging was investigated to detect fusarium damaged kernels (FDK) in Canadian wheat samples. More than 5,200 kernels, representing seven major Canadian wheat classes, with varying degree of infection symptoms ranging from sound through mild to severe were imaged in the visible-NIR (400–1,000 nm) wavelength range. Partial least squares discriminant analysis (PLS-DA) was used to segregate kernels into sound and damaged categories based on kernel mean spectra. A universal PLS-DA model based on four wavelengths was able to detect FDK in all seven classes with an overall accuracy of 90 % and false positives of 9 %.     

Early detection of toxigenic fungi on maize by hyperspectral imaging analysis

Fungi can grow on many food commodities. Some fungal species, such as Aspergillus flavus, Aspergillus parasiticus, Aspergillus niger and Fusarium spp., can produce, under suitable conditions, mycotoxins, secondary metabolites which are toxic for humans and animals. Toxigenic fungi are a real issue, especially for the cereal industry. The aim of this work is to carry out a non destructive, hyperspectral imaging-based method to detect toxigenic fungi on maize kernels, and to discriminate between healthy and diseased kernels. A desktop spectral scanner equipped with an imaging based spectrometer ImSpector- Specim V10, working in the visible-near infrared spectral range (400-1000 nm) was used. The results show that the hyperspectral imaging is able to rapidly discriminate commercial maize kernels infected with toxigenic fungi from uninfected controls when traditional methods are not yet effective: i.e. from 48 h after inoculation with A. niger or A. flavus.     

Maize populations with resistance to field contamination by aflatoxin B1

Maize (Zea mays L) germplasm is needed with resistance to infection by Aspergillus flavus and/or subsequent contamination by aflatoxin B₁ (AFB₁). A select group of maize populations were evaluated for their resistance to AFB₁ contamination at three locations. Four populations (Ibadan B and three others derived from crosses between Corn Belt inbreds and diploid perennial teosinte. Zea diploperennis) were compared with a susceptible hybrid check in a randomised complete block experiment with 10 replicates in Georgia, Missouri and South Carolina for two years. Ears were not inoculated, and naturally occurring concentrations of AFB₁ in harvested grain were analysed for population differences. Ibadan B and Mo2 W × Z diploperennis had significantly lower average amounts of AFB₁, 19 μg kg^?1 and 18 μg kg^?1, respectively, than other test entries. Backcrossing to susceptible Corn Belt inbreds produced populations as susceptible as the check when resistance was measured as the concentrations of AFB₁ in the grain. The consistency and significance of low AFB₁ concentrations for Ibadan B and Mo20W × Z diploperennis suggest that these may be useful sources of resistance.     

Distribution of fungi and aflatoxins in a stored peanut variety

The objective of the present study was to evaluate the mycoflora and occurrence of aflatoxins in stored peanut samples (hulls and kernels) from Tupã, State of São Paulo, Brazil. The samples were analyzed monthly over a period of one year. The results showed a predominance of Fusarium spp. (67.7% in hulls and 25.8% in kernels) and Aspergillus spp. (10.3% in hulls and 21.8% in kernels), and the presence of five other genera. The growth of Aspergillus flavus was mainly influenced by temperature and relative humidity. Analysis of hulls showed that 6.7% of the samples were contaminated with AFB₁ (mean levels = 15–23.9 μg/kg) and AFB₂ (mean levels = 3.3–5.6 μg/kg); in kernels, 33.3% of the samples were contaminated with AFB₁ (mean levels = 7.0–116 μg/kg) and 28.3% were contaminated with AFB₂ (mean levels = 3.3–45.5 μg/kg). Analysis of the toxigenic potential revealed that 93.8% of the A. flavus strains isolated were producers of AFB₁ and AFB₂.     

基于高光谱信息特征选择的玉米霉变程度Fisher鉴别方法

为了提高高光谱鉴别玉米霉变程度的正确率,分别对全波长和特征波长下霉变玉米进行鉴别分析。利用高光谱图像采集系统获得250个霉变玉米样本的高光谱数据,并用标准正态变量变换(standard normal variate,SNV)和多元散射校正(Multiplicative scatter correction,MSC)2种方法对原始数据进行预处理,再对未预处理和预处理后的原始数据进行判别,优选出多元散射校正的预处理方法;运用偏最小二乘回归系数选择了9个特征波长;运用Fisher判别分析(Fisher discriminant analysis,FDA)分别对全波长和特征波长下的训练集进行判别分析,并用对应的测试集进行检验。FDA结果表明,全波长下判别模型的训练集和测试集的准确率分别为97.71%,97.33%,9个特征波长下训练集和测试集的准确率分别为100.00%,98.67%。研究结果表明,利用特征光谱能够较好地表征玉米的霉变程度,有利于提高玉米霉变程度的鉴别正确率。