High-throughput Analysis of Aflatoxins in Cereals, Nuts and Processed Products Involving Automated Immunoaffinity Cleanup and Inline HPLC-Fluorescence Detection.
Background: The testing of aflatoxins (AFs) in fresh and processed foods is highly demanded to comply with trade regulations. Consequently, commercial laboratories face huge AF sample loads in food consignments. Worldwide, there is a rising interest to implement automation to increase sample throughput in AF analysis. Objective: This study sought to evaluate the performance of an automated cleanup and HPLC analysis system for determination of regulated AFs (B1, B2, G1, G2) in rice, flattened rice, sorghum, raw and processed peanut, almond, peanut butter, and wheat-based cookies. Methods: The samples were extracted with methanol-water (80:20), diluted with Triton X-100 and subjected to automated analysis, where the cleanup step through immunoaffinity column (IAC) and HPLC-fluorescence analyses [involving post-column bromination-derivatisation] were performed in 10 and 11 min, respectively. The method was validated in all test matrices at the LOQ and higher levels. The method performance was also evaluated against a conventional workflow where cleanup and HPLC analysis were manually performed. Results: The LOQ for peanut, sorghum, rice, and flattened rice was 0.125 ng/g, while it was 0.5 ng/g for peanut butter, almond, and wheat-based cookies. In all matrices, the recoveries at LOQ and higher levels were satisfactory. The double-cartridge exchange system completed the analysis of ∼96 injections in 18 h. Each IAC could be reused for 15-times, without incurring any recovery loss. The automated-system provided a better precision (RSD<9%) than the conventional (RSD=12-15%) workflow. Conclusions: Because of its high-throughput nature, this method is recommended for routine analysis of AFs. Highlights: A high-throughput method is reported where cleanup and HPLC analysis of aflatoxins were automatically performed. Each immunoaffinity column could be used 15-times without any loss in recoveries. The method performance was better than the conventional approach and complied with the analytical quality control guidelines. https://doi.org/10.1093/jaoacint/qsab083
Field Survival of the Brown Marmorated Stink Bug Halyomorpha halys (Hemiptera: Pentatomidae) on California Tree Crops.
Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) is a polyphagous insect pest that has invaded much of the United States, including California. Despite model predictions that regions in California like the agriculturally important Central Valley are suitable, H. halys populations and the occurring crop damage are distinctly lower than in regions of the East Coast. To evaluate if this is due to low nymphal survival in the high temperature and intensive monoculture environment, H. halys nymphs were caged on four common Central Valley tree crops: almond, pistachio, peach, and grape, and compared to the well-established almond and pistachio pest Leptoglossus zonatus (Dallas) (Hemiptera: Coreidae). Nymphal development showed that peach, almond, and pistachio can sustain H. halys as single host plants until adult eclosion, whereas grapes cannot. The addition of fruiting structures of almond, pistachio, and grape to H. halys caged on peach trees (= mixed diet) did not increase nymphal survival but did increase adult female size. Leptoglossus zonatus survival was higher than H. halys on pistachio, but not on the other crops. Overall, H. halys nymphal mortality in the field was high, with 92% in the mixed diet compared to 73% in the mixed diet control in the laboratory, indicating abiotic population pressures. Our results confirm peach as a good H. halys host plant and suggest that almond has a similar suitability, and while pistachios can support H. halys nymphal development, they seem less likely to facilitate population growth. https://doi.org/10.1093/ee/nvab055
Decontamination of aflatoxin B1 in peanuts using various cooking methods.
Peanut and its processed products are recurrently contaminated with aflatoxins (AFs) which are of potential public health concern. Among the different types of AFs, Aflatoxin B1 (B1) is the most frequently detected in peanuts over the maximum level (ML), and thus has warranted considerable research interest in the domain of food safety. In this study, we investigated the decontamination of B1 in three naturally-incurred lots (4, 12, and 40 µg/kg) of peanuts by a range of cooking treatments, including frying, pressure cooking, and roasting. B1 concentrations were determined by ultra-high performance liquid chromatography- fluorescence detection. The method provided a limit of quantification of 0.25 µg/kg for B1, which was much lower than any of its national and international MLs. The recoveries of B1 in fresh and cooked peanuts (positive-control) were in the range of 90-100%. Overall, all the cooking methods demonstrated a significant reduction in B1 loads. The degree to which the processing methods reduced the B1 content followed the pattern: roasting with a combination of NaCl and citric acid > pressure-cooking with a combination of NaCl and citric acid > frying. As the cooking procedures did not involve any complicated steps or sophisticated equipment, these could be readily adopted for decontamination or reduction in the level of B1 for a safer consumption of peanuts at the household level without affecting the organoleptic properties. https://doi.org/10.1007/s13197-020-04761-3
Effect of processing on polyphenol profile, aflatoxin concentration and allergenicity of peanuts.
Worldwide peanuts are often thermally processed before consumption, which might alter its bioactive composition and toxicity. The presented work explores the effect of processing methods such as roasting, frying, and pressure cooking on the bioactive composition, polyphenol profiles, aflatoxin concentration and, allergenicity response of peanuts. Effect of processing methods was assessed by monitoring total polyphenol content (TPC), total flavonoid content (TFC), antioxidant activity by DPPH & ABTS radicals, sensory analysis and, indirect ELISA. Compared to raw peanuts, all processing methods caused significant differences in TPC, TFC, DPPH & ABTS radical scavenging activities. Comparison of polyphenol profiles of raw to processed peanuts by LC-ESI-Q-TOF-MS chromatograms revealed different effects on concentrations of individual bioactive polyphenols. Indirect ELISA showed a significant decrease in the assay response on pressure cooking as compared to other samples. Total aflatoxin concentration was significantly reduced after processing in roasted (97.7%) and fried (98.3%) peanuts. Optimum processing conditions based on bioactive concentration and sensory analysis were found to be, roasting: 150 °C for 10 min, frying: 170 °C for 2 min and, pressure cooking: 15 min. Polyphenol profiles and bioactive constituents of peanuts were influenced by processing and may alter health benefits associated with them hence, vital for research and food industry applications. https://doi.org/10.1007/s13197-020-04779-7
A simple extraction method with no lipid removal for the determination of aflatoxins in almonds by liquid chromatography tandem-mass spectrometry (LC-MS/MS).
The present study describes a simple and rapid method for the determination of aflatoxins in almonds using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Aflatoxins were extracted using a modified QuEChERS method with little sample preparation, excluding the use of laborious purification procedures. Extracts were frozen overnight to separate the majority of lipids. The method was successfully validated for almonds. Linearity was demonstrated in the range 0.125-20 µg/kg. Limits of quantification (LOQ) ranged from 0.34 to 0.5 μg/kg. Matrix effect was not significant for the aflatoxins. Satisfactory recoveries were obtained at spike levels below 1 μg/kg and between 1 and 10 μg/kg. Relative standard deviations (RSDs) of repeatability and reproducibility were below 15%. The method was successfully tested with two proficiency tests in almond powder and peanut paste, with acceptable z-scores (-2 ≤ z ≤ 2). Only one of 11 local almond samples contained detectable aflatoxins, at concentrations below the maximum permitted level. https://doi.org/10.1080/19440049.2021.1925167
The Existing Methods and Novel Approaches in Mycotoxins’ Detection.
Mycotoxins represent a wide range of secondary, naturally occurring and practically unavoidable fungal metabolites. They contaminate various agricultural commodities like cereals, maize, peanuts, fruits, and feed at any stage in pre- or post-harvest conditions. Consumption of mycotoxin-contaminated food and feed can cause acute or chronic toxicity in human and animals. The risk that is posed to public health have prompted the need to develop methods of analysis and detection of mycotoxins in food products. Mycotoxins wide range of structural diversity, high chemical stability, and low concentrations in tested samples require robust, effective, and comprehensible detection methods. This review summarizes current methods, such as chromatographic and immunochemical techniques, as well as novel, alternative approaches like biosensors, electronic noses, or molecularly imprinted polymers that have been successfully applied in detection and identification of various mycotoxins in food commodities. In order to highlight the significance of sampling and sample treatment in the analytical process, these steps have been comprehensively described. https://doi.org/10.3390/molecules26133981
The Influence of the Addition of Nuts on the Thermal and Rheological Properties of Wheat Flour.
The aim of the study was to assess the influence of replacing wheat flour with hazelnuts or walnuts, in various amounts, on the thermal and rheological properties of the obtained systems. The research material were systems in which wheat flour was replaced with ground hazelnuts (H) or walnuts (W) in the amount of 5%, 10%, and 15%. The parameters of the thermodynamic gelatinization characteristics were determined by the differential scanning calorimetry method. In addition, the pasting characteristics were determined with the use of a viscosity analyzer and the viscoelastic properties were assessed. Sweep frequency and creep and recovery tests were used to assess the viscoelastic properties of the tested gels. It was found that replacing wheat flour with nuts increased the values of gelatinization temperature, gelatinization, and retrogradation enthalpy, and the degree of retrogradation. The highest viscosity was characteristic of the control sample (2039 mPa·s), and the lowest for the paste with 15% addition of walnuts (1120 mPa·s). Replacing the flour with nuts resulted in a very visible reduction in the viscosity of such systems. In addition, gels based on the systems with the addition of H and W were weak gels (tan δ = G″/G' > 0.1), and the values of G' and G″ parameters decreased with the increased share of nuts in the systems. Creep and recovery analysis indicated that the systems in which wheat flour was replaced with hazelnuts were less susceptible to deformation compared to the systems with the addition of W. https://doi.org/10.3390/molecules26133969
Environmental Conditions Affecting Ochratoxin A During Solar Drying of Grapes: The Case of Tunnel and Open Air-Drying.
Drying optimization, to mitigate fungal growth and Ochratoxin A (OTA) contamination is a key topic for raisin and currant production. Specific indicators of environmental conditions and drying properties were analyzed using two seedless grape varieties (Crimson-red and Thompson-white), artificially inoculated with Aspergillus carbonarius under open air and tunnel drying. The air temperature (T), relative humidity, grape surface temperature (Ts) and water activity throughout the drying experiment, the grapes' moisture content and the fungal colonization and OTA contamination during the drying process and their interactions were recorded and critically analyzed. Drying properties such as the water diffusivity (Deff) and peel resistance to water transfer were estimated. The grapes Ts was 5-7 °C higher in tunnel vs. open air-drying; the infected grapes had higher maximum Ts vs. the control (around 4-6 °C). OTA contamination was higher in tunnel vs. open air-dried grapes, but fungal colonies showed the opposite trend. The Deff was higher in tunnel than in the open air-drying by 54%; the infected grapes had more than 70% higher Deff than the control, differences explained by factors affecting the water transport. This study highlighted CFU and OTA indicators that affect the water availability between red and white grapes during open air and tunnel drying, estimated by the Deff and peel resistance. This raises new issues for future research. https://doi.org/10.3390/toxins13060400
Unraveling Genetic Diversity Amongst European Hazelnut (Corylus avellana L.) Varieties in Turkey.
European hazelnut (Corylus avellana) is a diploid (2n = 22), monecious and wind-pollinated species, extensively cultivated for its nuts. Turkey is the world-leading producer of hazelnut, supplying 70-80% of the world's export capacity. Hazelnut is mostly grown in the Black Sea Region, and maintained largely through clonal propagation. Understanding the genetic variation between hazelnut varieties, and defining variety-specific and disease resistance-associated alleles, would facilitate hazelnut breeding in Turkey. Widely grown varieties 'Karafındık' (2), 'Sarıfındık' (5), and 'Yomra' (2) were collected from Akçakoca in the west, while 'Tombul' (8), 'Çakıldak' (3), 'Mincane' (2), 'Allahverdi' (2), 'Sivri' (4), and 'Palaz' (5) were collected from Ordu and Giresun provinces in the east (numbers in parentheses indicate sample sizes for each variety). Powdery mildew resistant and susceptible hazelnut genotypes were collected from the field gene bank and heavily infected orchards in Giresun. Every individual was subjected to double digest restriction enzyme-associated DNA sequencing (ddRAD-seq) and a RADtag library was created. RADtags were aligned to the 'Tombul' reference genome, and Stacks software used to identify polymorphisms. 101 private and six common alleles from nine hazelnut varieties, four private from resistants and only one from susceptible were identified for diagnosis of either a certain hazelnut variety or powdery mildew resistance. Phylogenetic analysis and population structure calculations indicated that 'Mincane', 'Sarıfındık', 'Tombul', 'Çakıldak', and 'Palaz' were genetically close to each other; however, individuals within every varietal group were found in different sub-populations. Our findings indicated that years of clonal propagation of some preferred varieties across the Black Sea Region has resulted in admixed sub-populations and great genetic diversity within each variety. This impedes the development of a true breeding variety. For example, 'Tombul' is the most favored Turkish variety because of its high quality nuts, but an elite 'Tombul' line does not yet exist. This situation continues due to the lack of a breed protection program for commercially valuable hazelnut varieties. This study provides molecular markers suitable for establishing such a program. https://doi.org/10.3389/fpls.2021.661274
The HADES yield prediction system–a case study on the Turkish hazelnut sector.
Crop yield forecasting activities are essential to support decision making of farmers, private companies and public entities. While standard systems use georeferenced agro-climatic data as input to process-based simulation models, new trends entail the application of machine learning for yield prediction. In this paper we present HADES (HAzelnut yielD forEcaSt), a hazelnut yield prediction system, in which process-based modeling and machine learning techniques are hybridized and applied in Turkey. Official yields in the top hazelnut producing municipalities in 2004-2019 are used as reference data, whereas ground observations of phenology and weather data represent the main HADES inputs. A statistical analysis allows inferring the occurrence and magnitude of biennial bearing in official yields and is used to aid the calibration of a process-based hazelnut simulation model. Then, a Random Forest algorithm is deployed in regression mode using the outputs of the process-based model as predictors, together with information on hazelnut varieties, the presence of alternate bearing in the yield series, and agro-meteorological indicators. HADES predictive ability in calibration and validation was balanced, with relative root mean square error below 20%, and R2 and Nash-Sutcliffe modeling efficiency above 0.7 considering all municipalities together. HADES paves the way for a next-generation yield prediction system, to deliver timely and robust information and enhance the sustainability of the hazelnut sector across the globe. https://doi.org/10.3389/fpls.2021.665471