T-2 and HT-2 toxins are secondary metabolites of various species of Fusarium. These molecules can have high potential toxic effects for human and animal health. In this work, ELISA and ultra performance liquid chromatography with fluorescence detection (UPLC/FLD) were implemented and validated as screening and confirmatory tests for the detection of these two toxins in cereal samples. The developed methods were tested by analyzing 100 samples of cereals by ELISA screening for reducing costs and analysis time and then using UPLC/FLD for confirmation purposes. Both methods met the performance criteria for sensitivity, linearity, selectivity, precision, and ruggedness, as reported in the European Decision No. 2002/657/EC and in Regulation (EC) No. 401/2006. The correlation between ELISA and UPLC/FLD approaches showed good results (r = 0.9056), confirming that these two techniques should be considered to be complementary in the official control activities of cereal and derived products.
The occurrence of harmful cyanobacterial blooms in surface waters is often accompanied by the production of a variety of cyanotoxins, and these toxins are designed to target in humans specific organs on which they act. When introduced into the soil ecosystem by spray irrigation of crops, they may affect the same molecular pathways in plants having identical or similar target organs, tissues, cells, or biomolecules. There are also several indications that terrestrial plants, including crops, can bioaccumulate cyanotoxins and present, therefore, potential health hazards for humans. During this project, for monitoring purposes, water samples were collected from lake Occhito, in which there was an algal bloom (Planktothrix rubescens) in 2009, and from three tanks which acted as hydraulic junctions. In addition, crop samples irrigated with water from the three tanks mentioned above were also picked. Finally, the characterization of principal cyanobacteria was performed, to determine the presence of cyanotoxins such as microcystins and validate a method of screening ELISA for the determination of microcystins in vegetable samples and a confirmatory method by HPLC-ESI-MS/MS. Graphical abstract Occhito lake (left), microcystin LR (center), Tomato field in Foggia (right); figures below: ELISA (left), HPLC-MS/MS (right).
In this study, an Enzyme-Linked Immuno Assay (ELISA) was used to detect the presence of aflatoxin B1 (AFB1) and total aflatoxins (AFTs) in 200 samples of nuts for human consumption. The analytical technique was validated, with detection limit equal to 1 µg kg -1 and recovery percentage exceeding 90% for both AFB1 and AFT. The percentage of samples with aflatoxins concentration higher than legal limits amounted to 9% and 10% for AFB1 for AFT, respectively, and they were composed of almond and pistachio. Three samples of almond and 15 of pistachio resulted as non-complaint for AFB1. Other 2 samples of pistachio were non-compliant only for AFT. Taking into account that the levels exceeded up to five times the maximum permitted limits set by European Commission (AFT maximum concentration: 52.8 µg kg -1 ), the results of this survey confirmed the need of official control and the reliability of ELISA as sensitive screening method.
During a 5-year period from 2010 to 2014, n = 919 samples of feed and raw materials were analyzed for aflatoxin B (AFB) contamination using accredited ELISA screening methods. Only 0.76 % of these samples were non-compliant with maximum levels set by the European Union Regulation 32/2002. Non-compliant samples were mainly from the province of Bari (n = 3 samples, mean AFB value 7.03 μg/kg), although the highest AFB levels were found in two samples from the provinces of Foggia and Brindisi, at 32.6 ± 3.6 μg/kg and 31.0 ± 4.0 μg/kg, respectively. Mean AFB levels in samples contaminated but compliant with the limits ranged from 1.4 to 2.2 μg/kg. Considering the great importance of climate conditions in mycotoxins production, during crops production and during the critical phases of materials storage and/or transport, to better understand the variability in contamination levels, the analytical results were reviewed in term of temperature and relative environmental humidity in the sampling areas. Correlations between aflatoxin B levels in feed and these climate factors might explain seasonal and annual variations in contamination levels. The data from the present study provide useful suggestions for the organization of targeted monitoring plans and the protection of consumers, as well as for improvement in the quality standards of zootechnological activities and feed industry.
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