A simple and rapid biosensor for ochratoxin A based on a structure-switching signaling aptamer

A fast and reliable sensing platform has been developed for the detection of mycotoxin ochratoxin A (OTA) based on a target-induced structure-switching signaling aptamer. In the absence of target, a fluorescein-labeled OTA aptamer hybridizes to a complementary DNA strand containing a quencher moiety, bringing the fluorophore and the quencher into close proximity for highly efficient fluorescence quenching. Upon OTA addition, a conformational change in the aptamer releases the quencher-containing DNA strand, generating a strong concentration-dependent fluorescent signal. Using this technique, the entire detection and analysis process of OTA can be completed within 1 min. Under optimized assay conditions, a wide linear detection range (from 1 to 100 ng/mL) was achieved with a detection limit down to 0.8 ng/mL. Additionally, the proposed assay system exhibited high selectivity for OTA against other mycotoxins (aflatoxin B₁ and zearalenone) and limited interference from the structural analog ochratoxin B. The biosensor was also applied to a non-contaminated corn material spiked with a dilution series of OTA, obtaining recoveries from 83% to 106%. Utilization of the proposed biosensor for quantitative determination of mycotoxins in food samples may provide significant improvements in quality control of food safety through a simple, rapid, and sensitive testing system for agricultural products monitoring.     

Survey of aflatoxins and ochratoxin a contamination in food products imported in Italy

In this study the levels of aflatoxins (AF B₁, B₂, G₁ and G₂) and ochratoxin A (OTA) were monitored in several food products imported in Italy with a high contamination risk. A total of 345 samples were collected from the Maritime Authority of Salerno Customs Port during the period from January 2008 to December 2009 and analyzed by immunoaffinity chromatography as clean-up, high performance liquid chromatography with fluorescence detection for quantification and tandem mass spectrometry for confirmation. The analytical methods were validated on different food matrices and meet the performance criteria set by EC Regulation No. 401/2006 for mycotoxin analysis. The results obtained in this survey showed that 7% of the total samples contained detectable levels of AFs and OTA, and 1.2% had AFs concentrations exceeding the maximum limits set by EU regulation. OTA was the most prevalent mycotoxin, with an incidence of 17.6% of samples analyzed for OTA. The highest detected levels were 23.70 μg kg^?1 of OTA in a green coffee sample and 70.69 μg kg^?1 of AFs in an apricot kernels sample. Among the food products analyzed, hazelnuts paste and dried vine fruits were the commodities mainly contaminated with AFs and OTA, respectively.     

Natural occurrence of mycotoxins in cereals and spices commercialized in Morocco

Sixty samples of cereals (20 of corn, 20 of barley, and 20 of wheat) and 55 samples of spices (14 of paprika, 12 of ginger, 14 of cumin, and 15 of pepper) purchased from popular markets of Rabat and Salé in Morocco were analyzed for mycotoxins.Cereals samples were all analyzed for ochratoxin A (OTA). The average levels of contamination were 1.08, 0.42, and 0.17 μg/kg for corn, wheat, and barley, respectively. Samples of corn were also analyzed for zearalenone (ZEA) and fumonisin B₁ (FB₁) the average contaminations were 14 and 1930 μg/kg, respectively. Co-occurrence of OTA, FB₁, and ZEA was also checked. Spices samples were analyzed only for aflatoxins (AFs) and the average contaminations found for aflatoxin B₁ (AFB₁) were 0.09, 0.63, 2.88 and 0.03 μg/kg for black pepper, ginger, red paprika and cumin, respectively. The higher level of contamination was found in red paprika (9.68 μg/kg).The present report is the first one ever drafted on the natural co-occurrence of OTA, FB₁ and ZEA in cereals and on the occurrence of AFs in spices from Morocco.     

First survey on the presence of ochratoxin A and fungi in raw cereals and peanut available in the Republic of Niger

Eighty one (81) samples of cereals (rice, maize, sorghum, millet) and peanut purchased from six local markets of Niamey city (Republic of Niger) were analyzed for the presence of fungi and ochratoxin A (OTA). Samples were microbiologically analyzed for determination of fungi. For OTA analysis, samples were extracted with methanol with an Ultra-Turrax homogenizer. OTA was then identified and quantified by using liquid chromatography (LC) with fluorescence detection. The confirmation of OTA identity in positive samples was confirmed by methyl ester derivatization.Analytical results showed that samples were contaminated with a large number of fungi, the most important genus are Aspergillus spp. (63%) followed by Fusarium spp. (9.7%) and Penicillium spp. (3.3%). Mycotoxin analysis showed that OTA was present in 85.7, 9 and 2.6% of peanut, rice and maize, respectively. The frequency of contamination of total analyzed samples was 9.8%. Levels of OTA in positive samples ranged between 0.1 and 4.9 ng/g. The average contamination of cereals samples with OTA was 3.7 ng/g. The high level value of OTA (4.9 ng/g) was found in a peanut sample commercialized in “Wadata market” of Niamey city. The present paper is the first ever drafted on the natural occurrence of OTA and fungi in cereals and peanut consumed in Republic of Niger.     

Ochratoxin A in Spanish retail ground roasted coffee: Occurrence and assessment of the exposure in Catalonia

Occurrence of ochratoxin A (OTA) in ground roasted coffee from different brands and types available in Spain was assessed. Based on these data, exposure of the Catalan population to OTA through coffee consumption was estimated. Coffee samples were purchased in hypermarkets and supermarkets of twelve cities of Catalonia, Spain, and composite samples were prepared for analysis. OTA was extracted, cleaned-up by immunoaffinity columns, and detected by HPLC-fluorescence detection. Mean OTA content (n = 72) was 2.17 ± 0.79 ng/g (range 1.21–4.21 ng/g, 49% occurrence). An additional sampling was done by brands (n = 45), mean OTA contamination being 2.07 ± 0.61 ng/g (range 1.30–5.24 ng/g, 95% occurrence). Coffee consumption data were obtained by means of a food frequency questionnaire. Mean coffee consumption per capita was 11.58 ± 8.73 g/person/day. OTA daily intake (DI) was estimated by means of deterministic and probabilistic methods. In both cases, estimated DI (around 0.22 ng/kg bw/day) was below the latest PTDI value of 17 ng/kg bw/day suggested by EFSA.     

Antifungal activity mode of Aspergillus ochraceus by bacillomycin D and its inhibition of ochratoxin A (OTA) production in food samples

The inhibition effects of bacillomycin D on the growth of Aspergillus ochraceus and the production of ochratoxin A (OTA) in food samples were investigated. The mycelia growth and sporulation were completely inhibited by 30 μg/mL of bacillomycin D. Microscopic morphological changes such as the distortion of hyphae and the disruption of spores at 20 μg/mL of bacillomycin D were significantly observed. The use of bacillomycin D resulted in cell damage, nucleic acids and proteins divulge, and more production of reactive oxygen species (ROS), and all these factors actively contributed to the promotion of apoptosis of A. ochraceus. In addition, 90 μg/g of bacillomycin D completely inhibited the growth of A. ochraceus and the production of OTA in food samples. Our results suggested that bacillomycin D showed a significant antifungal activity against A. ochraceus that could be used as a potential natural antimicrobial to control food contamination and ensure food safety.     

Survey of aflatoxin B1 and ochratoxin A occurrence in traditional meat products coming from Croatian households and markets

The aim of this study was to investigate the occurrence of aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) in different traditional meat products circulating on Croatian markets, produced by a large number of households situated in different Croatian regions. The study involved a total of 410 samples of traditional pork meat products in terms of hams (n = 105), dry fermented sausages (n = 208), bacon (n = 62) and cooked sausages (n = 35), collected over four years period (2011–2014). Mycotoxin concentrations were quantified and confirmed using validated immunoassay method (ELISA) and high performance liquid chromatography with fluorescence detection (HPLC-FLD), respectively. The maximal observed OTA level in the fermented sausages and hams was around 5 times (5.10 μg/kg) to 10 times (9.95 μg/kg) higher than the maximal recommended level (1 μg/kg) stipulated for pork products in some EU countries. AFB₁ levels found in any given meat product analysed within this frame were not significantly higher (p > 0.05) than the applied method limit of detection. The results showed an occasional mycotoxin contamination of traditional meat products, especially that by OTA, pointing that to avoid such contamination meat and meat products on households should be produced and processed under standardised and well-controlled conditions.     

Aflatoxins and ochratoxin A in dried chestnuts and chestnut flour produced in Italy

The occurrence of aflatoxins and ochratoxin A in chestnut products made in Italy was surveyed. Thirty-seven samples of chestnut flour and fourteen of dried chestnuts were collected from retail outlets located in northern Italy. After extraction and purification through an immunoaffinity column, aflatoxins and ochratoxin A were analysed using both HPLC-FLD and HPLC-MS/MS. The mycotoxin contamination found in this survey was widespread and remarkable. As regards aflatoxins, the incidence of aflatoxin B₁ was 62.2 and 21.4% in chestnut flour and dried chestnuts, respectively; in the same products, the percentage of samples exceeding the value of 2.0 μg kg⁻¹ for aflatoxin B₁ (maximum limit fixed by EC Regulation 165/2010 in dried fruits) was 24.3% and 7.1%. A widespread and high incidence of AFG₁ was also found (40.5%). The maximum values for aflatoxin B₁ and total aflatoxins were 67.88 and 188.78 μg kg⁻¹, respectively (chestnut flour sample). Ochratoxin A occurred in all samples, showing very high values (mean 12.38 and 13.63 μg kg⁻¹ for chestnut flour and dried chestnuts, respectively); the percentages of samples exceeding the limit of 10 μg kg⁻¹ (EU limits for dried vine fruit) were 64.9 and 42.8% for chestnut flour and dried chestnuts, respectively. The maximum value was 65.84 μg kg⁻¹ (dried chestnut sample).     

Aflatoxin M1 and ochratoxin A in human milk in Ribeirão Preto-SP,Brazil

The objective of this study was to determine the extent of aflatoxin M₁ (AFM₁) and ochratoxin A (OTA) contamination in human breast milk in the city of Ribeirão Preto, São Paulo State, Brazil. During 2012, 100 samples of human milk were collected at the local Human Milk Bank. The method comprised, immunoaffinity column purification and isolation, liquid chromatography separation and fluorescence detection. The average percentage recoveries of AFM₁ and OTA spiked at 20 and 50 ng/L in control human milk were 78.1 ± 11.7% and 73.7 ± 9.6%, respectively. The average relative standard deviations of AFM₁ and OTA spiked at the same levels were 11.7 and 9.6% respectively. The limits of detection was 0.3 ng/L for AFM₁ and OTA. The limit of determination was 0.8 ng/L for both mycotoxins. This method was used to analyze 100 human milk samples, of which, two samples were found to contain AFM₁ at level greater than 0.3 ng/L. OTA was detected in 66 samples (66%), wherein 32 were above the limit of detection and 34 were in the range of from 0.8 to 21 ng/L. Results of our study indicate that breast-fed Brazilian infants had only an insignificant exposure to AFM₁ and OTA.     

Pressurized liquid extraction coupled to liquid chromatography for the analysis of ochratoxin A in breakfast and infants cereals from Morocco

A sensitive and reliable method using pressurized liquid extraction (PLE) and liquid chromatography (LC) has been developed for the analysis of ochratoxin A (OTA) in breakfast and infants cereals. Influence of several extraction solvents that affect PLE efficiency was studied. The selected PLE operating method was: 10 g of sample was packed into 22 ml stainless-steel cell and OTA was extracted with acetonitrile/water (80:20) at 40 °C, 34 atm in one cycle of 5 min at 60% flush. The mean recovery of OTA was 82 ± 4 at fortification level of 3 ng/g OTA. The limit of quantification (LOQ) of OTA was 0.25 ng/g. The proposed method was successfully applied to the analysis of 68 samples of breakfast and infants cereals products collected from different supermarkets and pharmacies in Rabat. Results showed that all analyzed infant cereals were free of OTA contamination. However, four samples of breakfast cereals were contaminated with OTA. Levels of OTA in positive samples ranged between 5.1 and 224.6 ng/g. All positive samples (5.8% of total samples) were above the maximum level set by EU regulations for OTA in cereal derivatives products.     

Co-occurrence of aflatoxins and ochratoxin A in spices commercialized in Italy

A total of 130 spice samples coming from India, China, South America, USA, Northern Africa, Europe and Sub-Saharan Africa were collected in different stores of Northern Italy. They were analysed for aflatoxins (AFs: AFB₁, AFB₂, AFG₁, AFG₂) and ochratoxin A (OTA) content by liquid chromatography with mass spectroscopy and positive electrospray ionization (LC/ESI-MS/MS), and HPLC with fluorescence detector (FLD), respectively. The analysis showed that 20 (15.4%) and 31 (23.8%) out of 130 samples were contaminated with AFs and OTA, respectively. A low level of total AFs contamination was found in the positive samples, the average concentration was 0.64 ng g⁻¹, far below the maximum threshold admitted by the European legislation (5 ng g⁻¹ for AFB₁, and 10 ng g⁻¹ for total aflatoxins (AFB₁, AFB₂, AFG₁ and AFG₂). A higher incidence of OTA was found in chili (60.0%) more than in pepper (13.3%), ranging from 2.16 to 16.35 ng g⁻¹, and from 1.61 to 15.85 ng g⁻¹, respectively. Moreover, three spice samples (2.3%) contaminated by OTA trespassed the threshold admitted by the European Regulation (EC, 2010). The co-occurrence of OTA and AFs in spices was detected in 6 out of 130 samples (4.6%), ranging from 1.61 to 15.85 ng g⁻¹ and from 0.57 to 3.19 ng g⁻¹, respectively.     

Natural occurrence of ochratoxin A in some marketed Nigerian foods

A total of one hundred and nine samples of maize (Zea mays) (17), millet (Pennisetum spp) (18), guinea corn (Sorghum) (17), acha (Digitaria exilis stapf) (20),sesame (Sesamum indicum) (19) and fermented cassava (Manihot esculenta) flakes (garri) (18) from markets located in Minna and its environs were analysed for ochratoxin A (OTA) by High Pressure Liquid Chromatography (HPLC). OTA was detected in 98.2% of the samples. The levels found were maize (0–139.2 μg/kg), millet (10.20–46.57 μg/kg), guinea corn (0–29.50 μg/kg), sesame seeds (1.90–15.66 μg/kg), acha (1.38–23.90 μg/kg) and garri (3.28–22.73 μg/kg). Maize had the highest level of OTA with “acha” having the lowest content of the toxin. The OTA levels found in marketed food and feed commodities which were mostly (74.3%) above 5 μg/kg, the European Union standard raise public health concern. The study is the first report of OTA contamination of acha, sesame seed and garri in Nigeria and possibly in Africa.     

A UPLC-MS/MS for simultaneous determination of aflatoxins, ochratoxin A, zearalenone, DON, fumonisins, T-2 toxin and HT-2 toxin, in cereals

An ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method is described for simultaneous determination of aflatoxins (AFB₁, AFB₂, AFG₁ and AFG₂), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisins (FB₁ and FB₂), T-2 and HT-2 toxins in cereals. Mycotoxins were separated by reverse phase liquid chromatography (RP-LC) and detected by tandem mass spectrometry using an electro spray-ionization interface (ESI) in both positive- and negative- ion modes. The mean recoveries of mycotoxins from spiked cereals ranged from 83.5% to 107.3%, whereas the limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.01 to 25 ng/g and 0.02-40 ng/g, respectively. The multi-mycotoxin method developed in this work was applied for the simultaneous determination of mycotoxins in 80 cereal samples collected from Malaysian markets. A total of 60 cereal samples (75%) were contaminated with at least one of these mycotoxins at levels greater than the LOD. Only one maize sample and two rice samples were contaminated at levels exceeding the European regulatory limits for aflatoxins and OTA (4 and 5 ng/g, respectively). The rates of the occurrence of mycotoxins in the commercial cereal samples were 50, 30, 19, 30, 16, 14, 14 and 12% for the aflatoxins (the total amount of AFB₁, AFB₂, AFG₁ and AFG₂), OTA, ZEA, DON, FB₁, FB₂, T-2 and HT-2 toxins, respectively. The results demonstrated that the procedure was suitable for the simultaneous determination of these mycotoxins in cereals and could be performed for their routine analysis in mycotoxin laboratories.     

Storage stability of maize-groundnut composite flours and an assessment of aflatoxin B1 and ochratoxin A contamination in flours and porridges

Defatting of groundnut flour used for composite development can not only improve nutritional quality of its products but also the storage stability. Maize, groundnut and their composite (full fat and defatted) flours were prepared and stored at room temperature over a period of 3 months. Storage stability of these products was assessed based on changes in water activity, peroxide value (PV), free fatty acids (FFA), thiobarbituric acid (TBA), microbiological profiling and levels of mycotoxins that included aflatoxin B₁ (AFB₁) and ochratoxin A (OTA). Overall results revealed that the rate of change of PV, FFA and TBA significantly (p ≤ 0.05) increased with increasing storage time, which was highest for full fat flours than in the defatted flours. For example, PV of the FFG and DFG were respectively, 0.88 and 0.40 mEq/kg, meanwhile TBA was 4.53 and 2.71 mg malonaldehyde/kg. There was a much higher rate of increase in FFA (%) with increasing storage time in full fat and composite flours when compared to that of their defatted counterparts. Generally, microbiological data demonstrated an increase in total microbial counts during storage in these foods possibly resulting in mycotoxins, AFB₁ (range: 9.08–38.48 μg/kg) and OTA (range: 0.33–19.50 μg/kg) in all samples with groundnut and maize having the highest contamination levels. A 127.8% increase in OTA level was noted when maize flour inclusion level in the full fat composite increased from 55 to 85%, but only a 24.7% increase in OTA level was noted in defatted composites during storage. Reducing the inclusion level of groundnut flour, the main source of AFB₁ as found, resulted in a drastic reduction in AFB₁ level in full fat and defatted composite flours by 54.1 and 76.4%, respectively, during storage. The findings highlight that shelf life stability of composites can be maintained upon defatting during the fortification process. It can therefore, be inferred that monitoring quality and safety of the raw materials as well as that of the final products during storage is crucial.     

Natural occurrence of aflatoxin B1, ochratoxin A and citrinin in Croatian fermented meat products

When domestic animals are exposed to mycotoxins, significant amounts of the latter shall be carried over into animal products such as milk, eggs and meat. This study was carried out in order to determine the possible presence of aflatoxin B₁ (AFB₁), ochratoxin A (OTA) and citrinin (CIT) in game sausages (n = 15), semi-dry sausages (n = 25) and fermented dry-meat products (n = 50), randomly taken from individual producers and the Croatian market. AFB₁ and OTA were quantified using ELISA, while CIT was quantified using HPLC-fluorescence detector. Out of 90 samples, the fungi most frequently isolated from dry-cured meat products were of Penicillium species, while Aspergillus was isolated from only one sample. As much as 68.88% of the samples were positive for mycotoxins. Finally, the analysis of different types of meat products resulted in OTA identification in 64.44%, CIT identification in 4.44% and AFB₁ identification in 10% of the samples. The maximum OTA concentrations established in the commercial sausage samples equalled to 7.83 μg/kg, while that of AFB₁ amounted to 3.0 μg/kg. Generally, although OTA was detected in all three types of products in different percentage shares, mutual differences were not statistically significant (P > 0.05).     

Cross-linked chitosan polymers as generic adsorbents for simultaneous adsorption of multiple mycotoxins

The primary objective of this study was to synthesize three types of cross-linked chitosan polymers and further investigate their adsorption capability for multiple mycotoxins, including aflatoxin B₁ (AFB₁), ochratoxin A (OTA), zearalenone (ZEN), fumonisin B₁ (FB₁), deoxynivalenol (DON) and T-2 toxin (T2). Among these synthetic adsorbents, cross-linked chitosan-glutaraldehyde complex presented the highest adsorption capability for AFB₁ (73%), OTA (97%), ZEN (94%) and FB₁ (99%), but no obvious adsorption for DON and T2 (<30%). The effect of various incubation conditions (contact time, dosage and pH) was also studied. Subsequently, the experimental data were fitted to Langmuir, Freundlich and Hill models. The best fitting model to describe AFB₁ and FB₁ adsorption was Langmuir model (R² ≥ 0.99), with the theoretical maximum adsorption amounts of 5.67 mg/g for AFB₁ and 15.7 mg/g for FB₁. The Hill model was the best model for OTA and ZEN adsorption (R² > 0.98), with the predicted maximum adsorption amounts were 24.8 mg/g for OTA and 9.18 mg/g for ZEN. In addition, the adsorption capability of adsorbent for the simultaneous presence of multiple mycotoxins was also evaluated in buffer system and simulated gastrointestinal condition. The results indicated that the coexisted multiple mycotoxins didn't affected the adsorption capability of adsorbent, whereas the adsorption amounts of toxins were decreased by some gastrointestinal components. The findings of this research suggest that chitosan–glutaraldehyde complex has the potential to be applied as multitoxin adsorbent material for reducing the combined adverse effect of multiple mycotoxins on humans and animals.     

An ultrasensitive gray-imaging-based quantitative immunochromatographic detection method for fumonisin B1 in agricultural products

Fumonisins (FBs) are widely found in rice, maize, peanut, wheat, and other agricultural products. These have been detected using a chromatography technique, whereas the rapid assay by a highly sensitive monoclonal antibody is minimally reported. Herein, a highly sensitive monoclonal antibody (7A11) was successfully developed by hybridoma technique. The 50% inhibition concentration (IC₅₀) of 7A11 monoclonal antibody was 0.32 ng/mL in an optimized buffer. The cross-reactivity between fumonisin B₁ and fumonisin B₂, B₃ was 4.3% and 12.8%, respectively. Based on the newly developed 7A11 antibody, a high sensitivity indirect competitive enzyme-linked immunosorbent assay (icELISA) and gold nanoparticles-based gray imaging quantification immunoassay (GNPs-GI) were established. The analytical range of icELISA was 0.08–1.38 ng/mL and that for GNPs-GI was 0.24–15 ng/mL. Both the methods showed adequate recoveries (80.0–105.8% for icELISA and 78.5–115.2% for GNPs-GI) in spiked samples. Compared to high-performance liquid chromatography (HPLC), icELISA and GNPs-GI indicated reliability that could be used for further detection of fumonisin B₁ in agricultural products.     

Estimation of dietary intakes of fumonisins B1 and B2 from conventional and organic corn

The dietary intakes of fumonisins from 60 samples of conventional and organic corn were assessed. A 13.3% of the conventional corn samples contained fumonisin B₁ and B₂ at mean levels of 43 and 22 ng/g, respectively, while 10% of the organic corn samples contained fumonisins at somewhat lower levels of 35 ng/g (FB₁) and 19 ng/g (FB₂). Overall, the fumonisin levels in the corn samples were much lower than the maximum level of 2000 ng/g (as the sum of FB₁ and FB₂) proposed for unprocessed maize in a recent EU regulation. The fumonisins present in conventional and organic maize are estimated to contribute with very low percentages of 0.21% and 0.17%, respectively, to the level considered at risk for human health. Based on the data exposed in this paper, the farming system is probably not of decisive importance for the final contamination of agricultural products with these mycotoxins.     

Effect of gamma radiation on reduction of mycotoxins in black pepper

Gamma ray was applied to reduce mycotoxins, i.e. ochratoxin A (OTA) and aflatoxins B₁, B₂, G₁ and G₂ (AFB₁, AFB₂, AFG₁ and AFG₂) in black pepper. Response surface methodology (RSM) was applied to evaluate the effect of dose of gamma ray ranging from 0 to 60 kGy and mycotoxin concentration ranging from 10 to 100 ng g⁻¹ on the mycotoxin reduction. The maximum reduction was found at 60 kGy which was 52%, 43%, 24%, 40% and 36% for OTA, AFB₁, AFB₂, AFG₁ and AFG₂, respectively. Results showed the gamma rays even at 60 kGy were not effective in completely destroying of ochratoxin and aflatoxins.     

Ochratoxin A determination in paired kidneys and muscle samples from swines slaughtered in southern Italy

A survey on 54 paired kidneys and muscle samples from pigs slaughtered in Southern Italy, has been carried out in order to check the presence of ochratoxin A.Both competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) and high-performance liquid chromatography with fluorescence detection (HPLC-FD) were used to determine ochratoxin levels. Mean and median value for OTA in kidneys were 0.29 and 0.25 ng/g, respectively. Mean and median values found for muscle (0.024 and 0.01 ng/g) are significantly lower than those reported in other studies demonstrating that, irrespective of the geographical provenance of pigs, OTA incidence is far from representing a real concern for consumers.Results obtained plotting ELISA vs. HPLC results show that ELISA tends to slightly underestimate the OTA content compared to HPLC; nevertheless, ELISA remains an invaluable tool as rapid screening semiquantitative technique.