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Deoxynivalenol (DON), a highly prevalent food contaminant, is known to induce reproductive and immunotoxicity in humans upon exposure. The present study focused on the consequences of exposure to DON during pregnancy for placental barrier and immune function, as well as fetal survival. Female mice received diets contaminated with DON (6.25 and 12.5 mg/kg of diet), starting immediately after mating until the end of the experiment. On day 17 of pregnancy the animals were killed, and maternal and fetal samples were collected for further analysis. Feeding on DON-contaminated diets decreased fetal survival, and DON was detected at significant levels in the fetus. Placentae from DON-exposed mice revealed a reduction in expression of junctional proteins, ZO-1, E-cadherin and claudins, upregulation of AHR mRNA expressions, and increase in IFN-ꝩ, IL-6 and IL-4 production. In conclusion, results of this study demonstrate harmful effects of DON on the course of pregnancy and fetal survival, which might be due to immunological changes in maternal immune organs and placenta. Altogether, these data underline the importance of the quality of maternal diet during pregnancy as they clearly demonstrate the potential harmful effects of a commonly present food-contaminant.

This study investigated effects of dietary fumonisins (FBs) on gut and faecal microbiota of weaned pigs. In total, 18 7-week-old male pigs were fed either 0, 15 or 30 mg FBs (FB1 + FB2 + FB3)/kg diet for 21 days. The microbiota was analysed with amplicon sequencing of the 16S rRNA gene V3-V4 regions (Illumina MiSeq). Results showed no treatment effect (p > 0.05) on growth performance, serum reduced glutathione, glutathione peroxidase and malondialdehyde. FBs increased serum aspartate transaminase, gamma glutamyl-transferase and alkaline phosphatase activities. A 30 mg/kg FBs treatment shifted microbial population in the duodenum and ileum to lower levels (compared to control (p < 0.05)) of the families Campylobacteraceae and Clostridiaceae, respectively, as well as the genera Alloprevotella, Campylobacter and Lachnospiraceae Incertae Sedis (duodenum), Turicibacter (jejunum), and Clostridium sensu stricto 1 (ileum). Faecal microbiota had higher levels of the Erysipelotrichaceae and Ruminococcaceae families and Solobacterium, Faecalibacterium, Anaerofilum, Ruminococcus, Subdoligranulum, Pseudobutyrivibrio, Coprococcus and Roseburia genera in the 30 mg/kg FBs compared to control and/or to the 15 mg/kg FBs diets. Lactobacillus was more abundant in the duodenum compared to faeces in all treatment groups (p < 0.01). Overall, the 30 mg/kg FBs diet altered the pig gut microbiota without suppressing animal growth performance.

Fusarium fujikuroi species complex (FFSC) strains are a major concern for food quantity and quality due to their strong ability to synthesize mycotoxins. The effects of interacting conditions of water activity, temperature, and incubation time on the growth rate, toxin production, and expression level of biosynthetic genes were examined. High temperature and water availability increased fungal growth. Higher water activity was in favor of toxin accumulation. The maximum amounts of fusaric acid (FA) and fumonisin B1 (FB1) were usually observed at 20–25 °C. F. andiyazi could produce a higher content of moniliformin (MON) in the cool environment than F. fujikuroi. The expression profile of biosynthetic genes under environmental conditions varied wildly; it was suggested that these genes might be expressed in a strain-dependent manner. FB1 concentration was positively related to the expression of FUM1, while a similar correlation of FUB8 and FUB12 with FA production could be observed in F. andiyazi, F. fujikuroi, and F. subglutinans. This study provides useful information in the monitoring and prevention of such toxins entering the maize production chain.

The contamination of fermented feeds and foods with fungi and mycotoxins is a major food safety issue worldwide. Certain lactic acid bacteria (LAB), generally recognized as safe (GRAS) fermentation probiotics, are able to reduce microbial and mycotoxins contamination. In this study, Lactiplantibacillus (L.) plantarum Q1-2 and L. salivarius Q27-2 with antifungal properties were screened as inoculants for mixed fermenting feed, and the fermentation and nutritional qualities, microbial community, and mycotoxins of mixed fermented feed were analyzed at different fermentation periods (1, 3, 7, 15, and 30 days, respectively). The findings indicated that the utilization of Q1-2 and Q27-2 strains in fermenting feed led to a decrease in pH and an increase in lactic acid concentration and the proportion of Lactiplantibacillus, while effectively restraining the proliferation of undesirable microorganisms. In particular, Q1-2 reduced the relative abundance of fungi including Fusarium and Aspergillus. Compared to the control group, the Q1-2 and Q27-2 groups reduced aflatoxin B1 by 34.17% and 16.57%, and deoxynivalenol by up to 90.61% and 51.03%. In short, these two LAB inoculants could reduce the contents of aflatoxin B1 and deoxynivalenol to the limited content levels stipulated by the Chinese National Standard GB 13078-2017. These findings suggest that the LAB strains of Q1-2 and Q27-2 have potential applications in the feed industry for the mitigation of mycotoxin pollution, thereby enhancing the quality of animal feed.

An inter-laboratory collaborative study was performed to evaluate the performance of the Biochip Array Technology (BAT) Myco 7 method. The Myco 7 Array is a method which simultaneously and quantitatively detects 20 mycotoxins including aflatoxins B1, B2, G1 and G2, ochratoxin A, deoxynivalenol, zearalenone, fumonisin B1, B2 and B3 and T-2 and HT-2 toxin. The BAT Myco 7 method was collaboratively evaluated by nine government and private Association of American Feed Control Officials (AAFCO) laboratories. Samples were analysed in a proficiency testing round format. Seventeen blind samples were analysed on the same equipment using Myco 7 kits. 99% of the results fell within an acceptable Z-score range of -2|1, B2, G1 and G2 assays. Fumonisins and ochratoxin A assays had a 93% and 94% pass, respectively. The reproducibility co-efficiency of variation was between 16 and 20% meeting set criterion of

Chillies and chilli-based products are important spices on a global basis. The production, processing, transport and storage phases of chillies are prone to infection by Aspergillus Section Flavi and contamination with aflatoxins (AFs), especially aflatoxin B1 (AFB1) for which legislative limits exist in many countries. We have examined the effect of the interacting abiotic factors of water availability (water activity, aw; 0.995-0.850 aw) and temperature (15-37 °C) on (a) lag phases prior to growth, (b) growth, (c) AFB1 production and (d) contour maps of optimum and boundary conditions for colonisation and toxin production by three Aspergillus flavus strains on a 10% chilli-based medium. Additional studies with whole red chillies + A. flavus conidial inoculum on AFB1 contamination during storage for 10-20 days at 30 °C were also carried out. In vitro, the lag phases before growth were delayed by lower temperatures (15, 20 °C) and aw levels (0.928-0.901 aw). There was no statistical difference in growth between the three strains. Optimal growth was at 37 °C and 0.982 aw with no growth at 0.85 aw. Optimal temperature × aw conditions for AFB1 production were at 30 °C and 0.982 aw with no statistical difference in production between strains. No AFB1 was produced at 15-20 °C at 0.901 and 0.928 aw levels, respectively. In situ studies with A. flavus inoculated whole red chillies at 0.90 and 0.95 aw found that this species became the major component of the total fungal populations at 30 °C after 10-20 days storage. AFB1 contamination was above the European legislative limits (5 μg/kg) for spices at 0.90 aw after 20 days storage and at 0.95 aw after 10 and 20 days. This suggests that storage conditions of ≥0.90 aw, especially at ≥25-30 °C represents a significant risk of contamination with AFB1 at levels where rejection might occur, even after only 10-20 days storage.

Maize is consumed world-wide as staple food, livestock feed, and industrial raw material. However, it is susceptible to fungal attack and at risk of aflatoxin contamination under certain conditions. Such contamination is a serious threat to human and animal health. Ensuring that the maize used by food industry meets standards for aflatoxin levels requires significant investment across the supply chain. Good Agricultural Practices (GAP) form a critical part of a broader, integrated strategy for reduction of aflatoxin contamination. We reviewed and summarised the GAP of maize that would be effective and practicable for aflatoxin control within high-risk regions for smallholder farmers. The suggested practicable GAP for smallholder farmers were: use of drought-tolerant varieties; timely harvesting before physiological maturity; sorting to remove damaged ears and those having poor husk covering; drying properly to 13% moisture content; storage in suitable conditions to keep the crop clean and under condition with minimally proper aeration, or ideally under hermetic conditions. This information is intended to provide guidance for maize growers that will help reduce aflatoxin in high-risk regions, with a specific focus on smallholder farmers. Following the proposed guidelines would contribute to the reduction of aflatoxin contamination during pre-harvest, harvest, and post-harvest stages of the maize value chain.

Aflatoxin M1 (AFM1) contamination in milk and milk products may pose a major public health concern. The present cross-sectional study was aimed to estimate the prevalence of AFM1 in bovine milk across all districts of Punjab, India and to identify the associated animal and farm level risk factors. A total of 402 milk samples (266 cow milk and 136 buffalo milk) were analysed using commercial ELISA and representative samples were confirmed using HPLC-FLD. The results revealed that 56.2 and 13.4% of the milk samples exceeded the maximum levels of the European Union, i.e. 0.05 μg/l and Food Safety and Standards Authority of India (FSSAI), i.e. 0.5 μg/l for AFM1 in milk, respectively. On analysis of species variation, buffalo milk (prevalence: 56.6%; mean concentration: 0.42±0.9 μg/l) was found to have higher AFM1 levels than cow milk (prevalence: 56.0%; mean concentration: 0.19±0.3 μg/l), with statistically significant difference between mean concentrations (P1 prevalence (P=0.91). Furthermore, milk from commercial dairy farms (prevalence: 64.7%; mean concentration: 0.34±0.65 μg/l) was found to be more contaminated than from household dairy establishments (prevalence: 47.8%; mean concentration: 0.19±0.65 μg/l). The risk factors ‘above average milk yield/day’ (odds ratio (OR): 2.4) and ‘poor animal hygiene’ (OR: 1.9) were identified at animal level, and ‘intensive dairy farming’ (OR: 3.1) and ‘animal feed without aflatoxin binder’ (OR: 4.7) as farm level risk factors for AFM1 excretion above maximum levels of European Union in milk. Among cow breeds, the milk from ‘non-descript’ breed (OR: 11.5) was found to be most contaminated with AFM1 and the least from Jersey breed (OR: 1.0). The present study highlighted the presence of AFM1 in milk samples; therefore, regular monitoring of AFM1 in milk is required so that high risk regions and associated risk factors can be addressed appropriately.

Deoxynivalenol (DON) is a mycotoxin that commonly contaminates cereals worldwide. Dietary exposure to DON is a subject of great public health concern, but studies on the health effects of chronic exposure to DON are not available. In this study, we investigated the connection between DNA methylation levels and DON exposure in children. The DNA methylation status of white blood cells from 32 children aged 2~15 years old in Henan, China, was profiled. A total of 378 differentially methylated CpGs were identified between the high and low DON exposure groups, and 8 KEGG pathways were found to be significantly enriched among the differentially methylated genes. In addition, the quantitative methylation of EIF2AK4, EMID2 and GNASAS was analysed using the Sequenom MassARRAY platform. The results showed that the methylation level of EIF2AK4 was significantly different between the two groups, and the methylation levels were associated with exposure to DON. Conclusively, our study found that chronic exposure to DON during childhood could affect DNA methylation levels.

The need for a healthy market in the rapid and accurate screening of a variety of pathogenic agents and toxins in the environment and food has led to an increase in the development of new biosensors, which have ideal characteristics, such as high sensitivity and specificity with rapid detection and simple preparation of the sample. Among the food contaminants, mycotoxins have been identified as a major challenge for the food industry, and rapid and accurate detection has attracted the attention of food inspection and monitoring organisations. In this study, a nanomolecular detection method is described using aflatoxin B1 (AFB1)-specific aptamers attached to streptavidin-coated magnetic nanoparticles. A prominent feature of the AFB1-specific aptamers is a guanine-rich (G-rich) sequence with a G-quadruplex structure after capturing AFB1 molecules and mimicking peroxidase activity. The enzymatic reaction evaluated in the presence of chromogenic substrate and measurement is done by a smartphone-specific application for colorimetric measurement. The results indicated that the assay could measure AFB1 in rice, flour, seed, maize, and pistachio. In addition, the application of hybrid nanomaterial technology resulting from the binding of biotin-labelled aptamers to the surface of streptavidin-coated magnetic nanoparticles minimises preparation and treatment of samples, improves results, and consequently reduces false negative and positive responses in the detection field. This study may eventually lead to the design and development of a fast, sensitive, specific, and on-site AFB1-based nanomolecular colorimetric detection system via a smartphone-based application that can be readily accessible to all applicants, from professionals to manufacturers of foodstuffs.

Fumonisins (FBs), a widespread group of mycotoxins produced by Fusarium spp., are natural contaminants in cereals and foodstuffs. Fumonisin B1 (FB1) is the most toxic and prevalent mycotoxin of this group, and it has been reported that FB1 accounts for 70-80% of FBs produced by the mycotoxigenic strains. The mode of action of FB1 depends on the structural similarity with sphinganine/sphingosine N-acyltransferase. This fact causes an accumulation of sphingoid bases and blocks the sphingolipid biosynthesis or the function of sphingolipids. Diverse toxic effects and diseases such as hepatocarcinogenicity, hepatotoxicity, nephrotoxicity, and cytotoxicity have been reported, and diseases like leukoencephalomalacia in horses and pulmonary oedema in horses and swine have been described. In humans, FBs have been associated with oesophageal cancer, liver cancer, neural tube defects, and infantile growth delay. However, despite the International Agency for Research on Cancer designated FB1 as a possibly carcinogenic to humans, its genotoxicity and epigenetic properties have not been clearly elucidated. This review aims to summarise the progress in research about the genotoxic and epigenetics effects of FB1.

Grain contamination by mycotoxins can cause significantly negative health and economic impact in areas where poor agricultural practices and food insecurity is prevalent. This study was conducted to investigate the current postharvest practices and aflatoxin contamination awareness level amongst maize producers in Jimma Zone, Ethiopia. Semi-structured questionnaires were used for quantitative and qualitative data collection from 90 randomly selected maize producers in two districts through a cross-sectional study design. The study revealed poor postharvest practices due to lack of proper infrastructure. Maize is harvested after maturation with the use of traditional sun drying. Plastic sheets are commonly used to protect harvested maize from rain during field drying. The majority of the respondents reported the use of traditional storage structures (81.1%) with great potential for possible mould proliferation and aflatoxin production. Maize producers of up to 62.2% were ignorant about aflatoxins and up to 26.7% reported the possible usage of aflatoxin contaminated maize in human food preparation and animal feed production. Up to 53.3% of the respondents were ignorant of aflatoxin risks in human health and stability during food processing hence the usage of mouldy maize for tella brewing. Further investigations on aflatoxin levels in maize-based food and beverages, and human exposure studies are needed. Moreover, there is a need to enhance the maize producers’ knowledge on good agricultural practices and mycotoxicosis through awareness programmes.

The objective of this study was to evaluate the potential intra-species variability of 3 Fusarium langsethiae strains in response to extreme climate change (CC) conditions on an oat-based matrix. The impact of elevated temperature (25 vs 30-34 °C) coupled with increasing drought stress (0.98 vs 0.95 aw) and elevated CO2 (400 vs 1000 ppm) were examined on lag phases prior to growth, growth rate, and production of the mycotoxins T-2 and HT-2 and their ratio. In comparison to the control conditions (25 °C; 0.98; 400 ppm), exposure to increased temperature (30-34 °C), showed similar reductions in the lag phase and fungal growth rates of all 3 strains. However, with elevated CO2 a reduction in both lag phases prior to growth and growth rate occurred regardless of the aw examined. For T-2 and HT-2 mycotoxin production, T-2 showed the most intra-species variability in response to the interacting abiotic stress factors, with the 3 strains having different environmental conditions for triggering increases in T-2 production: Strain 1 produced higher T-2 toxin at 25 °C, while Strain 2 and the type strain (Fl201059) produced most at 0.98 aw/30 °C. Only Strain 2 showed a reduction in toxin production when exposed to elevated CO2. HT-2 production was higher at 25 °C for the type strain and higher at 30-34 °C for the other two strains, regardless of the aw or CO2 level examined. The HT-2/T-2 ratio showed no significant differences due to the imposed interacting CC abiotic conditions.

Multi-mycotoxin exposure data are missing to guide risk assessment and legislation in Ethiopia. This study therefore aimed to determine mycotoxin contamination levels in maize samples from 176 randomly selected household storages in three agro ecological zones of South (Sidama zone) and West (Jimma zone) Ethiopia, and to examine the post-harvest practices and household processing. Liquid chromatography coupled to tandem mass spectrometry was used to quantify 23 mycotoxins. The majority of the households regularly consumed maize (more than once per day). More (77%) samples in Sidama were contaminated with Fusarium mycotoxins deoxynivalenol than in Jimma (29%) (P1 (19%), fumonisin B2 (19%) and fumonisin B3 (12%, Sidama vs 13%, Jimma) contamination were observed (P>0.05). In Sidama, only one sample was contaminated with the Aspergillus mycotoxin aflatoxin B2 and another sample with aflatoxin B1. From all samples, 40% were contaminated with 3-5 types of Fusarium mycotoxins and only 4% of the samples were contaminated with 6-8 types of mycotoxins. After the harvested maize was dried on the field, the majority of respondents in Jimma reported that they removed the maize within one day, which was less practiced in Sidama. The majority of households in Sidama, and some in Jimma, reported that they dried maize before storage, mainly using the sun. Close to two third of the study participants in the two zones reported that they applied the chemical dichloro-diphenyl-trichloroethane (DDT) during maize storage. All households in both zones reported that they sorted visible mouldy maize grains before preparation of maize flour while most of them kept the mouldy maize for animal feed. Protective strategies of Fusarium mycotoxin contamination, with special focus on deoxynivalenol and zearalenone, should be well promoted in the study areas as they are possible human and animal health threats.

Patulin (PAT) is a toxic mycotoxin usually contaminated apple juices, which leads to a serious food safety issue in the world. Thiol-compounds are a class of compounds containing the thiol (-SH) group themselves or obtained the -SH group by physical or chemical modification. They have the ability to efficiently remove patulin in apple juices with manifested negligible effects on juice quality. This review investigates the latest development in the removal of patulin using thiol-compounds, including the removal efficiencies and mechanisms of patulin, the factors influencing the removal efficiency of patulin, as well as the toxicities of thiol-compounds and safety of juices after detoxification. This review shows that thiol-compounds are promising materials for the removal or degradation of patulin in the contaminated juices.

The study investigated the short-term effects of a single oral bolus of zearalenone (ZEN) on the rumen microbiota and fermentation patterns in four rumen-cannulated Holstein cows fed a forage diet with daily 2 kg/cow concentrate. During the baseline day, cows received uncontaminated concentrate, followed by ZEN-contaminated concentrate on the second day, and again the uncontaminated concentrate on day three. Free rumen liquid (FRL) and particle-associated rumen liquid (PARL) were collected at different hours post-feeding on all days to analyze the prokaryotic community composition, absolute abundances of bacteria, archaea, protozoa, and anaerobic fungi, as well as short-chain fatty acid (SCFA) profiles. The ZEN reduced the microbial diversity in FRL but not in the PARL fraction. The abundance of protozoa was higher after ZEN exposure in PARL, which may be related to their strong biodegradation capacity that, therefore, promoted protozoal growth. In contrast, α-zearalenol might compromise anaerobic fungi as indicated by reduced abundances in FRL and fairly negative correlations in both fractions. Total SCFA significantly increased in both fractions after ZEN exposure, while the SCFA profile only changed marginally. Concluding, a single ZEN challenge caused changes in the rumen ecosystem soon after intake, including ruminal eukaryotes, that should be the subject of future studies.

Rice is a widely consumed food worldwide; however, it can be a source of pollutants, such as potentially toxic elements (PTEs), mycotoxins, and pesticides. Sixty rice samples imported from Pakistan (PAK), India (IND), and Thailand (THAI), as well as domestic Iranian (IRN) rice, were collected from Bushehr, Iran, and investigated for the contamination of PTEs, including arsenic (As), lead (Pb), cadmium (Cd), and nickel (Ni); pesticides, including chlorpyrifos, trichlorfon, diazinon, fenitrothion, and chlorothalonil; mycotoxins, such as aflatoxin B1 (AFB1), zearalenone (ZEN), ochratoxin A (OTA), and deoxynivalenol (DON); and molds. Estimated daily intake (EDI) and hazard quotient (HQ) of pollutants and hazard index (HI) and incremental lifetime cancer risk (ILCR) of rice types for the Iranian adult population were calculated. The content of PTEs in Iranian rice was not higher than Iran’s national standard limits. In contrast, other types of rice (imported) had at least one PTE above the permissible level. OTA content was below the detection limit, and all other mycotoxins were within the allowable range in all rice types. Thai rice was the only group without pesticides. The HI order of rice types was as follows: HIPAK = 2.1 > HIIND = 1.86 > HIIRN = 1.01 > HITHAI = 0.98. As was the biggest contributor to the HI of Iranian and Thai rice, and diazinon in the HI of Pakistani and Indian rice. The calculation of ILCR confirmed that the concentrations of Ni and Pb in Pakistani and Ni and As in Indian, Thai, and Iranian rice were not acceptable in terms of lifetime carcinogenic health risks.

Aflatoxin B1 (AFB1) is a mycotoxin known to impair human and animal health. It is also believed to have a deleterious effect on ruminal nutrient digestibility under in vitro batch culture systems. The objective of this study was to evaluate the effects of increasing the dose of AFB1 on ruminal dry matter and nutrient digestibility, fermentation profile, and N flows using a dual-flow continuous culture system fed a diet formulated for lactating dairy cows. Eight fermenter vessels were used in a replicated 4 × 4 Latin square design with 10 d periods (7 d adaptation and 3 d sample collection). Treatments were randomly applied to fermenters on diet DM basis: (1) 0 μg of AFB1/kg of DM (Control); (2) 50 μg of AFB1/kg of DM (AF50); (3) 100 μg of AFB1/kg of DM (AF100); and (4) 150 μg of AFB1/kg of DM (AF150). Treatments did not affect nutrient digestibility, fermentation, and N flows. Aflatoxin B1 concentration in ruminal fluid increased with dose but decreased to undetectable levels after 4 h post-dosing. In conclusion, adding incremental doses of AFB1 did not affect ruminal fermentation, digestibility of nutrients, and N flows in a dual-flow continuous culture system fed diets formulated for lactating dairy cows.

Global efforts to feed the burgeoning population have focused on increasing food production and access. In the meantime, FAO estimates about 1/3 food loss in the developing countries. To minimize these losses, grain storage techniques were investigated in an earthquake hit village in Kavre district of Nepal. Pesticide free moisture proof/airtight food storage bags were provided to 1,055 households using financial support from UNICEF-Nepal. Thirty-three households were identified who could store food grains for 6-month for this study. Maize and rice grains were stored in porous and in moisture-proof hermetic bags. Food grain samples were collected at the beginning and at 6-month and were analyzed for physical quality parameters, nutrients and mycotoxins. About 92% insect damage occurred in maize stored in porous bags which was prevented in triple layer hermetic Purdue Improved Cowpea Storage (PICS) bags. Insect damage in paddy remained low within 10% in both treatments. Major maize nutrient loss occurred through insect damage. The traditional practice was to use insect and mold infested maize as feed. Mycotoxins results showed that toxigenic molds develop in open storage of improperly dried grains especially maize. Fusarium-related mycotoxins associated with stunting were detected in maize samples using LC-MS/MS equipment. Mature grains should be dried sooner to processing moisture contents and packaged into moisture-proof hermetic bags (Dry Chain) to minimize postharvest loss. A clear policy implication is that use of improved low-cost Dry Chain technology should be promoted to enable quality food systems.

Dried fig is one of the most susceptible products to aflatoxin contamination. Since contaminated figs are not suitable for human consumption and cannot be used for any other purposes, they are burned in a chemical incinerator. In this study, we investigated the potential of using aflatoxin-contaminated dried figs as a raw material for ethanol production. For this purpose, contaminated dried figs (and also uncontaminated controls) were subjected to fermentation and subsequent distillation, and the alcohol and aflatoxin levels were determined during the processes. In addition, volatile by-products in the final product were determined using gas chromatography. Contaminated and uncontaminated figs had similar fermentation and distillation patterns. Although fermentation caused significant decreases in aflatoxin levels, there were still toxin residues in the fermented samples at the end of the process. On the other hand, aflatoxins were completely removed in the first step of the distillation. There were minor differences between the volatile compound compositions of the distillates produced from contaminated and uncontaminated figs. It was shown that obtaining aflatoxin-free and high-alcohol-content product using contaminated dried figs is possible according to the lab-scale conducted studies. Aflatoxin-contaminated dried figs can be used as a sustainable raw material for producing ethyl alcohol that can be used as an ingredient of surface disinfectants and/or fuel additive for vehicles.

In recent years, the growth of the field of medicinal herbs and the increased demand for plant-derived products have raised questions about their safety for consumer health. The evaluation of mycotoxins and pesticide residues in these matrices are among the major concerns. For this reason, forty-seven samples of different medicinal herbs were purchased from Italian herbal shops and the presence of these contaminants was assessed. AFB1, AFB2, AFG1, AFG2 and OTA A were characterised using high-performance chromatography with a FLD detector (LOQs were set at 0.375 μg/kg for aflatoxins and 1.0 μg/kg for ochratoxin A). A QuEChERS modified method was used for pesticide residue determination followed by GC–MS/MS and LC–MS/MS analysis (LOQ was set at 0.005 mg/kg). The two strategies proved to be complementary and contributed to a greater pesticide residues characterisation. About thirty percent of the samples analysed were found to be contaminated. Data highlighted the importance and the need to implement international monitoring in these types of matrices.

Citrinin (CIT), a mycotoxin known to exert nephrotoxicity, is a contaminant in food and feed. Since CIT contamination is not regularly analyzed, data on its occurrence and especially levels in food commodities are insufficient for conducting a conventional exposure assessment. Yet, human biomonitoring, i.e., an analysis of CIT and its metabolite dihydrocitrinone (DH-CIT) in urine samples allows to estimate exposure. This study investigated CIT exposure in young (2–14 years) and adult (24–61 years) residents of three federal states in Germany. A total of 179 urine samples from children and 142 from adults were collected and analyzed by a targeted LC-MS/MS based method for presence of CIT and DH-CIT. At least one of the biomarkers was detected and quantified in all urines, which indicated a widespread dietary exposure to the mycotoxin in Germany. Interestingly, the biomarker concentrations of CITtotal (sum of CIT and DH-CIT) were higher in children’s urine (range 0.05–7.62 ng/mL; median of 0.54 ng/mL) than in urines from adults (range 0.04–3.5 ng/mL; median 0.3 ng/mL). The biomarker levels (CITtotal) of individual urines served to calculate the probable daily CIT intake, for comparison to a value of 0.2 µg/kg bw/day defined as ‘level of no concern for nephrotoxicity’ by the European Food Safety Authority. The median exposure of German adults was 0.013 µg/kg b.w., with only one urine donor exceeding this provisional tolerable daily intake (pTDI) for CIT. The median exposure of children was 0.05 µg/kg bw per day (i.e., 25% of the pTDI); however, CIT exposure in 12 individuals (6.3% of our study group) exceeded the limit value, with a maximum intake of 0.46 µg/kg b.w. per day. In conclusion, these results show evidence for non-negligible exposure to CIT in some individuals in Germany, mainly in children. Therefore, further biomonitoring studies and investigations aimed to identify the major sources of CIT exposure in food commodities are required.

Mycotoxins constitute a real problem of public health because of their highly toxic effects for the populations. This study aims to evaluate the practices that favor aflatoxins and ochratoxin A development in rice sold on the markets of Abidjan (Cote d’Ivoire). A food consumption survey was conducted among 45 traders and 135 consumers of rice in three markets in Abobo and Cocody communes. A simple, open-ended questionnaire was submitted to the rice traders and consumers. It covered socio-demographic characteristics (age, sex, level of education), location and conditions of storage, place of supply, and knowledge of mycotoxins. Following the survey, 20 samples of rice, prized by the population, were collected and analyzed. Total aflatoxin and ochratoxin A levels were determined using HPLC coupled with mass spectrometry. The results revealed that all traders were unaware of the existence of mycotoxins. The storage and marketing of rice are housed in very unhygienic premises (lack of brick walls in some cases, poor quality tin roof, premises not swept regularly). Most traders renew their rice stocks over an average of one to two months (66.66% in Abobo; 79.99% in Cocody) and prefer to buy their supplies from large stores and other markets in Abidjan. Aflatoxin B1 (0.75±0.05 µg/kg) and total aflatoxin (1.73±0.34 µg/kg) were detected in the rice samples from Abobo market. The presence of Aflatoxins in rice could be related to poor post-harvest practices identified. Key words: Rice, total aflatoxins, ochratoxin A, practices, Abidjan.

Bacillus spp. have been used as a biocontrol strategy to eliminate/reduce toxic fungicides in viticulture. Furthermore, the presence of fungi that are resistant to commonly used products is frequent, highlighting the need for new biocontrol strains. Aspergillus carbonarius can produce ochratoxins, including ochratoxin A (OTA), which has a regulatory maximum allowable limit for grape products. The purpose of this study was to assess the ability of four Amazonian strains of Bacillus (P1, P7, P11, and P45) to biocontrol A. carbonarius and various forms of ochratoxins in grapes. Berries treated with strain P1 presented no fungal colonies (100% reduction), while P7, P11 and P45 strains caused a reduction of 95, 95 and 61% on fungal counts, respectively. Six forms of ochratoxin were found in the grapes inoculated with A. carbonarius, including ochratoxin ��, ochratoxin ��, ochratoxin �� methyl-ester, ochratoxin �� amide, N-formyl-ochratoxin �� amide, and OTA. Four of these ochratoxin forms (ochratoxin ��, ochratoxin �� methyl-ester, ochratoxin �� amide, N-formyl-ochratoxin �� amide) are reported for the first time in grapes. These ochratoxins were identified using liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QToF-MS). All Bacillus strains inhibited the synthesis of OTA, which is the most toxic form of ochratoxin. No ochratoxin form was found when P1 and P7 were used. Although some forms of ochratoxin were detected in grapes treated with Bacillus spp. P11 and P45, the levels decreased by 97%. To our knowledge, this is the first report on the inhibition of Aspergillus carbonarius-derived ochratoxin by Bacillus species. P1 strain, identified as Bacillus velezensis, was found to be the most promising for completely inhibiting fungal growth and production of all ochratoxins.

Mycotoxins are frequently found in fruits and fruit juices. However, data about occurrence and fate of mycotoxins along the fruit juice processing chain are currently insufficient. Herein, a liquid chromatographic/tandem mass spectrometric (LC–MS/MS) multi-mycotoxin method was developed and applied to investigate the effect of technological unit operations on the fate of three of the most relevant mycotoxins along the processing chain for cloudy and clear apple juice, namely patulin (PAT), ochratoxin A (OTA), and alternariol (AOH). Raw juice obtained directly after dejuicing was spiked with the aforementioned mycotoxins at pilot-plant scale prior to subjecting it to different technological unit operations. Regarding clear apple juice production treatment with a pectinolytic enzyme preparation, and pasteurization were insignificant for mycotoxin reduction, but fining with subsequent filtration was effective, although the mycotoxins showed different affinity towards the tested agents. The most effective fining agent was activated charcoal/bentonite in combination with ultrafiltration, which removed OTA (54 µg/L) and AOH (79 µg/L) to not quantifiable amounts (limit of quantification (LOQ) 1.4 and 4.6 µg/L, respectively), while PAT was reduced only by 20% (from 396 to 318 µg/L). Regarding cloudy apple juice production, all studied processing steps such as centrifugation and pasteurization were ineffective in reducing mycotoxin levels. In brief, none of the common steps of clear and cloudy apple juice production represented a fully effective safety step for minimizing or even eliminating common mycotoxins. Thus, ensuring the sole use of sound apples should be of utmost importance for processors, particularly for those manufacturing cloudy juices.

A major challenge in post-harvest protection of chilies is its rapid drying to prevent quality deterioration as a result of fungal contamination. The speed and efficiency of drying is crucial as the improper and prolonged drying may eventually initiate the buildup of mycotoxins. This research demonstrated levels of aflatoxin in samples at different stages of drying with the objective to assess the efficiency of the common practice of on-farm sun drying in Pakistan. The results indicated that there was no significant pre-harvest contamination of aflatoxins in chilies grown at different locations under study. Sun drying of the harvested chilies over a post drying stage reduced average moisture content from 69.70% to 9.87%, but also led to a gradual increase in the level of aflatoxins. The correlation between the observed increase in aflatoxin levels and the length of the drying period was found to be statistically significant (P

Mycotoxins are important food contaminants posing a significant threat to food and feed safety and public health. Among the mycotoxins, aflatoxins are deemed to be a more significant contaminant due to their potent carcinogenic, and hepatotoxic effects, and their levels are highly regulated in the international food trade. Phytochemicals are considered a major source of natural antifungal agents. The volatile nature of essential oil of plants makes them ideal candidates for antifungal agents due to their ability to distribute in free air spaces in closed containers and penetrate through heterogeneous food materials. In these, essential oils in spices attain special attention due to their commercial availability and low toxicity. This article reviews the antiaflatoxigenic capacity of spice essential oils and the effect of essential oil composition on the activity and mechanism of antifungal action and is expected to be useful for the planning of further research in the subject area.

In Ontario, Canada, Fusarium graminearum Schwabe causes Gibberella ear rot (GER) in maize, resulting in the accumulation of mycotoxins, mainly deoxynivalenol (DON), DON-3-glucoside (DON-3G) and zearalenone (ZEN) in infected kernels. Fungicides can be an important tool for managing GER and DON and other Fusarium mycotoxins in maize. Until recently, all fungicides available to growers were triazoles, thus no resistance management strategy through fungicide use was possible. In this study, a novel carboxamide fungicide active ingredient (pydiflumetofen) was evaluated against conventional triazole fungicides and mixtures for: (1) effectiveness on mycotoxins (2) optimal application timing; and (3) efficacy of application, with and without an insecticide, under natural and inoculated-misted conditions. The best timing for fungicide application was at full silk, resulting in the highest reduction of GER symptoms and lowest accumulation of F. graminearum mycotoxins in harvested grain. DON and DON-3G concentrations were reduced by at least 50% with a fungicide application at full silk. Fungicide treatments did not affect fumonisin concentrations in grain. Pydiflumetofen (94 g active ingredients (AI)/ha) and fungicides containing pydiflumetofen (75-94 g AI/ha) were similar to standard triazole fungicides (prothioconazole at 200 g AI/ha and metconazole at 90 g AI/ha) for reducing GER and F. graminearum mycotoxins under misted-inoculated plots and commercial field conditions; as a result, we expect pydiflumetofen to be competitive with triazole-only chemistries in the marketplace, which should delay the onset of fungicide resistance.

Mycotoxins are natural food contaminants, associated with adverse health effects due to acute intoxication and, what is much more common, chronic exposure. The most hazardous and widespread mycotoxins are subjected to regulation in food in most countries. Alongside with regulated, a wide list of mycotoxins is monitored in various foods. Traditionally mycotoxins in tea are determined in a dry sample, not taking into account their transfer rate into the infusion. This study was aimed at the determination of the transfer rate of several mycotoxins from naturally contaminated herbal tea samples into an infusion. Seven of the most contaminated samples were pre-selected during the monitoring of mycotoxins in Camellia sinensis and herbal tea available in the Russian Federation. Ochratoxin A (OTA), sterigmatocystin (STE), mycophenolic acid (MPA), tentoxin (TTX), alternariol (AOH), its methyl ether (AME), zearalenone (ZEN), enniatins A and B (ENN A and B), beauvericin (BEA) were detected in these samples in the range of several μg/kg to several mg/kg. HPLC-MS/MS was used for dry tea samples and their infusion analysis. Mycotoxin polarity and infusion pH (for analytes possessing carboxylic groups) appeared to be factors determining transfer rate. STE transferred into infusion at the average rate of 10%. Average transfer of Alternaria toxins varied from 73% (TTX) to 45% (AOH) and about 11% (AME). A third part of ZEN was detected in the infusion. Transfer of ENNs and BEA was low and did not exceed 7%. Infusion pH affected MPA transfer rate; it increased from 23% to 96% in the pH range from 5.5 to 6.3. 83% of OTA was detected in the infusion of a single contaminated sample. Consideration of the mycotoxin transfer rate to herbal tea infusions resulted in the change of the model herbal tea input into mean chronic dietary exposure for most studied mycotoxins.

Fusarium head blight (FHB) causes yield loss, quality reduction, and grain mycotoxin accumulations. A novel pydiflumetofen-containing fungicide, Miravis Ace, was recently registered in North America. The main objective of this study was to assess the efficacies of Miravis Ace and the timing of application alongside industry standard triazole fungicides (Prosaro, Caramba, Proline and Folicur) on suppressing FHB, reducing mycotoxins and improving wheat agronomic performance. The assessment was conducted across six natural environments on commercial farm fields and in two artificially inoculated-misted environments. All environments included 5 fungicides (Miravis Ace and the four triazole fungicides) and 3 application timings (Zadoks GS 59, 65, 69-71). Additionally, for the ZGS 65 timing, the experiment in the natural environment included a quinone outside inhibitor (QoI) fungicide pyraclostrobin (Headline). In general, Miravis Ace tended to be more effective on FHB suppression than the triazole fungicides across all environments. However, any biological differences tended to be statistically non-significant, likely because of a lack of statistical power. Miravis Ace reduced total deoxynivalenol (DON) concentration by 52-73% compared to the non-treated control. If applied at ZGS 59-65, Miravis Ace was more effective in increasing yield and test weight than the triazoles tested. Across fungicides, applications made at ZGS 65 were most effective in FHB suppression compared to earlier or later application timings. There was no evidence that pyraclostrobin increased mycotoxin concentrations. Overall, compared to the triazole fungicides, the novel pydiflumetofen-containing fungicide tended to have lower FHB suppression and mycotoxins, higher grain yield and test weight, and higher harvest moisture, but differences were not always statistically significant. Because the main active ingredient in Miravis Ace has a different mode of action than the triazoles, we speculate that this fungicide will be competitive with industry standards, and benefit strategies for fungicide resistance management.

A protective effect of two herbs, Glycyrrhiza glabra and Tinospora cordifolia, given as feed additives was observed against the growth inhibitory effect of ochratoxin A (OTA) and associated immunosuppression and biochemical or pathomorphological changes. The feed levels of 3 mg/kg OTA and fine powder of one of both herbs were given during a period of 32 days to female broiler chicks divided into 3 experimental and 1 control groups (14 chicks per group). The observed pathological and biochemical changes, the changes in relative organs’ weight and body weight, and the decrease of antibody titer against Newcastle disease were more pronounced in the OTA-treated chicks without herbal supplementation, and less pronounced in the chicks treated additionally with G. glabra or T. cordifolia as was shown by the better feed performance and the higher body weight in the chicks treated with the herbs. The higher relative weight of lymphoid organs of the chicks supplemented with both herbs revealed their beneficial effects on the immune system. The hepatoprotective effect of both herbs was evident, being stronger in the chicks additionally supplemented with G. glabra shown by the pathomorphological findings and by the lower levels of aspartate transaminase (131.1 U/l) compared to chicks given only OTA (156.0 U/l). A protective effect of T. cordifolia on the bone marrow and kidneys was found as was shown by the lower levels of uric acid (382.9 μmol/l) compared to chicks given only OTA (466.9 μmol/l).

In a temperate climate, the mycotoxins deoxynivalenol (DON), nivalenol (NIV), enniatin B (ENN B), mycophenolic acid (MPA), roquefortine C (ROC) and zearalenone (ZEN) are often found in maize silage. Although rumen microbiota are able to degrade some mycotoxins (e.g. DON), others are known to stay mainly intact (e.g. ROC). In addition, mycotoxin degradation can be hampered by a low ruminal pH or decrease in rumen microbial activity. Hence, these mycotoxins can reach the small intestine and exert a cytotoxic effect on intestinal epithelial cells. In this study, a real-time kinetic analysis of the cytotoxicity of these six mycotoxins and some of their metabolites (α- and β-zearalenol, α- and β-ZOL) was performed in a calf small intestinal epithelial cell line (CIEB). Confluency as well as the cell death parameters apoptosis and necrosis were determined to evaluate the mycotoxin-induced cytotoxicity. A combination of Annexin-V green and Cytotox red staining was used to determine early and late apoptosis as well as necrosis. Six different concentrations were tested ranging from 0.78 to 12.5 μM. Compared to cells not exposed to mycotoxins, DON and NIV exert a fast toxic effect with DON being more toxic than NIV within the first hours of incubation, whereas the inverse was observed at 16 h of incubation. On the other hand, MPA and ZEN induced increased Annexin V green positive cells within several hours of incubation with higher toxicity over time. Increased Annexin V green and Cytotox red positive cells were seen for ROC only at the highest concentration tested. For ENN B, increased Annexin V green positive cells were observed only after 12 h and α- and β-ZOL did not show cytotoxic effects. Hence, mycotoxin exposure causes either severe (DON and NIV) or more limited (ZEN, ROC, MPA, and ENN B) risk of bovine intestinal epithelial damage.

Aflatoxins are toxic and carcinogenic secondary metabolites, produced by Aspergillus flavus and Aspergillus parasiticus, which contaminate food and feed and threaten human and animal health. To assess the prevalence of aflatoxins in Tanzania, 180 groundnut and 200 maize samples were collected from 9 and 10 districts, respectively. Aflatoxin contamination was quantified using high performance liquid chromatography. Aflatoxins were detected in samples collected from all districts and prevalence ranged from 92 to 100% for groundnuts and 10 to 80% for maize. The mean aflatoxin level for groundnuts was 6.37 μg/kg and the highly contaminated sample had 40.31 μg/kg. For maize, the mean aflatoxin level was 12.47 μg/kg and the highly contaminated sample had 162.40 μg/kg. The estimated average probable daily intake (APDI) of aflatoxin B1 (AFB1) from groundnuts consumption was 1.88 ng/kg body weight/day, while for maize, it ranged between 151.98-272.89 ng/kg body weight/day. The APDI for both groundnut and maize exceeded the provisional maximum tolerable daily intake (PMTDI) of AFB1 for adults (1 ng/kg body weight/day), bringing about health concerns for populations in Tanzania. Another alarming finding was that 75% of the farmers who provided samples for analysis were not aware of aflatoxins or the negative health impacts from consuming contaminated products. Results reported in this paper show that aflatoxin contaminated staple crops are widely distributed in Tanzania and that the risk of human exposure is high due to diet preferences. Awareness campaigns are required to inform and protect farmers and consumers.

In this study, we report a new method to prepare highly sensitive monoclonal antibody against α-zearalanol (ZAL) based on a similar antigen determinant structure. Zearalanone (ZAN), structural analogs of ZAL, was modified by oximation to obtain ZAN-O. ZAN-O was then coupled with bovine serum albumin using 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) to synthesise the artificial complete antigen ZAN-O-BSA. ZAN-O-BSA was used to immunise the BALB/c mice. The splenocytes of the immunised mice were fused with myeloma NS0 cells. During the process of cell fusion, ZAL was used as an inhibitor instead of ZAN to screen the hybridoma cell lines that can secrete monoclonal antibodies against ZAL. The sensitivity (half inhibitory concentration, IC50) of the prepared monoclonal antibody was 0.475 ng/ml, the limit of detection (LOD) was 0.050 ng/ml, the linear range of detection was 0.066-3.399 ng/ml, the affinity constant Kaff was 6.18×107 l/mol, the cross-reactivity rate with structural analogues, such as β-zearalanol, α-zearalenol, β-zearalenol, ZAN and zearalenone were 28.07, 13.16, 15.83, 60.28 and 7.95% respectively. The cross-reactivity with other mycotoxin and carrier proteins were all less than 0.05%. The prepared monoclonal antibody can be used to establish a highly sensitive immunoassay for the detection of ZAL.

Tea is one of the most popular beverage in the world and may be contaminated by fungi and mycotoxins during processing. To analyse aflatoxin B1 (AFB1) and sterigmatocystin (STC) in three types of t...

The contamination of foods and feeds with mycotoxins has been an issue of global significance. For mycotoxin detoxification, enzymatic biodegradation using laccase has received much attention. In this study, a laccase gene lac2 from the fungus Pleurotus pulmonarius was expressed in the Pichia pastoris X33 yeast strain to produce recombinant proteins. Enzymatic properties of recombinant Lac2 and its ability to degrade zearalenone (ZEN) and Aflatoxin B1 (AFB1) in the presence of four mediators (ABTS, TEMPO, AS and SA) were investigated. Result showed that the optimum pH and temperature of recombinant Lac2 were 3.5 and 55 °C, respectively. Lac2 was not sensitive to heat and stable under both acidic and alkaline conditions. Lac2-ABTS and Lac2-AS were efficient systems for ZEN degradation over a wide range of pH (4–8) and temperature (40–60 °C). Lac2-AS was the most efficient system for AFB1 degradation, reaching 99.82% of degradation at pH 7 and 37 °C after 1 h of incubation. Finally, the Lac2-mediator oxidation products were structurally characterized. This study lays a solid foundation for the application of Lac2 laccase combined with AS for degrading mycotoxin in food and feed.

New information is needed regarding the types and concentrations of mycotoxins in swine feed. We hypothesized that (1) the mycotoxins deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON), nivalenol (NIV), and zearalenone (ZEN) vary among swine ingredient and feed types, and (2) the inclusion of specific ingredients is associated with mycotoxin contamination in complete feed. A total of 707 samples were collected from cooperators in 14 states between June 2018 and January 2020 then analyzed for DON, 3-AcDON, 15-AcDON, NIV, and ZEN contamination using gas chromatography–mass spectrometry (GC–MS). Ninety-four percent (663/707) of samples contained DON, 33% (230/707) of samples contained 3-AcDON, 57% (404/707) of samples contained 15-AcDON, 1% (6/707) of samples contained NIV, and 47% (335/707) of samples contained ZEN. Seventy-three percent (514/707) of samples contained multiple mycotoxins. Resulting DON concentrations were below the national advisory limits for all sample types, and no advisory limits are imposed for the other mycotoxins studied. Increased incorporation of distiller’s dried grains with solubles (DDGS) was associated with increased DON in complete feed (R2 = 0.82).

The objective of this study was to evaluate biochemical parameters and histopathology of liver in Matrinxã (Brycon cephalus) and Pacu (Piaractus mesopotamicus) fish chronically exposed to dietary aflatoxins. Fish feed was artificially contaminated with aflatoxins and the treatments were: Control – feed without toxin; Treatment A – feed + 10 μg aflatoxin B1 (AFB1)/kg; Treatment B – feed + 20 μg AFB1/kg; and Treatment C – feed + 50 μg AFB1/kg. Matrinxã and Pacu juvenile fish were placed in tanks for 180 days. Five experimental units per treatment were monthly sampled and submitted to blood collection and removal of hepatic tissue. Thus, twenty blood and liver samples for each species were collected monthly, adding up to 240 samples analysed. To verify biochemical changes, analyses included total proteins, albumin, globulins, aspartate aminotransferase (AST) and alkaline phosphatase (ALP). The hepatic tissue was examined microscopically and the slides presenting histopathological changes were photo-documented. There was effect of treatment (P0.05) was observed in Pacu. A reduction (P1.

The objective of this work was to evaluate the aflatoxins residues on tissues of Pacu (Piaractus mesopotamicus) fish chronically exposed to dietary aflatoxins, also evaluating the effects on fish performance. Aflatoxins were incorporated into the extruded fish feed and the concentrations were confirmed by high performance liquid chromatography (HPLC). The experimental design comprised the following treatments: Control – feed without toxin; Treatment A: feed + 10 μg aflatoxin B1 (AFB1)/kg; Treatment B: feed + 20 μg AFB1/kg and Treatment C: feed + 50 μg AFB1/kg. Pacu juvenile were allocated in tanks with density of one fish per litter. The experiment lasted 180 days with monthly sampling, and approximately ten fish per treatment were used to compose a sample. Biometric surveys were performed every 30 days, including standard weight and length. Aflatoxins were detected and quantified in fish muscle and liver by HPLC, using immunoafinity columns. Results showed AFB1 deposition in the liver throughout the experiment in all treatments, but only treatment C differed significantly (P1 residues at low levels in liver and only a slight amount in fish muscle, it should be noted that accumulation is possible in the Pacu species. The diets contaminated with AFB1 negatively interfered in weight and length of fish when compared to control. Therefore, it can be concluded that long-term exposure of Pacu to dietary AFB1 can cause losses to producers.

Plant-based beverages (popularly known as vegetable milk) have become increasingly important in recent years. However, the nonexistence of information on mycotoxin contamination is noticeable. We herein describe the development and validation of an analytical methodology that employs QuEChERS and LC-MS/MS for the simultaneous determination of nine mycotoxins (aflatoxins B1, B2, G1, and G2, fumonisins B1 and B2, ochratoxin A, zearalenone, and citreoviridin) in seven types of vegetable milk (peanut, oat, rice, cashew, maize, soybean, and coconut). The method provided the following quantification limits, recoveries at the lowest validated concentration and relative standard deviations under repeatability conditions at the lowest validated concentration, respectively: aflatoxin B1 (0.023 μg/l, 84.98 and 9.23%); aflatoxin B2 (0.024 μg/, 93.00 and 4.85%); aflatoxin G1 (0.057 μg/l, 98.85 and 5.53%); aflatoxin G2 (0.031 μg/l, 96.64 and 4.08%); fumonisin B1 (2.166 μg/l, 75.55 and 16.78%); fumonisin B2 (1.105 μg/l, 70.47 and 11.89%); ochratoxin A (0.104 μg/l, 72.05 and 5.12%); zearalenone (8.093 μg/l, 107.10 and 6.37%); citreoviridin (1.305 μg/l, 97.25 and 7.28%). The method uses small amounts of samples, solvents, and other inexpensive reagents with no need for laborious clean-up and pre-concentration steps. Its attractive characteristics (simplicity, low cost compared to procedures that use immunoaffinity columns, and full compatibility with routine analyses) make it potentially valuable. As a proof-of-principle, the validated methodology was applied to seven commercial samples of different compositions showing that some were contaminated with aflatoxins and ochratoxin A.

The inclusion of anti-mycotoxin additives (AMA) in the diet of production animals has been widely used to avoid mycotoxin exposure. In order to confirm the efficacy of such products in vivo, measurement of mycotoxins and/or their metabolites in biological fluids is preconized. This study aimed at determining the serological biomarkers of zearalenone (ZEN), α-zearalenol, β-zearalenol, α-zearalanol, β-zearalanol (β-ZAL) and zearalanone, to evaluate the efficacy of an AMA in beef heifers. The trial lasted 37 days: 11 days of adaptation, 21 days of actual experiment, and 5 days of regression. Twenty-four heifers were randomly assigned to receive one of the following treatments (n=6/group): (T1) basal diet (control); (T2) basal diet + 5 mg/kg of ZEN; (T3) basal diet + 5 mg/kg of ZEN + 2.5 kg/t of AMA; and (T4) basal diet + 5 mg/kg of ZEN + 5.0 kg/t of AMA. Blood sampling was performed on different days after the diet was given. The samples were centrifuged to obtain the blood serum, and then analysed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). β-ZAL was detected above the limit of quantification both in the unconjugated (>0.60 ng/ml) and conjugated (>0.90 ng/ml) forms. The remaining metabolites presented concentrations under the limit of detection. In the efficacy evaluation of the AMA, there was no significant difference (P>0.05) between the treatments with and without additive at the tested levels of inclusion. Thus, β-ZAL may be employed as a biomarker of ZEN exposure via diet to evaluate the efficacy of an AMA through serological parameters. The technique applied in this study proved to be an adequate alternative for in vivo confirmation of the efficacy of products in adsorbing the toxin.

Bentonite clays exhibit high adsorptive capacity for contaminants and is frequently used as a feed additive to reduce the bioavailability and thus the toxicity of several mycotoxins. Zearalenone (ZEN) is a secondary Fusarium toxic metabolite that can contaminate a wide range of food- and feedstuff. Since organophilic treatments is known to increase the adsorption capacity of bentonites, the aim of study was to evaluate and compare the ability of natural bentonite and bentonite treated with BAC302 to protect against ZEN-induced cytotoxicity in the epithelial colorectal adenocarcinoma (Caco-2) and human leukemia monocytic (THP-1) cell lines. The two materials were not toxic to the cell lines at lower concentrations. Furthermore, the results indicate that the two materials protect the Caco-2 and THP-1 cells against ZEN-induced cytotoxicity, probably by extracellular adsorption of ZEN. The tested natural bentonite shows potential for in vivo testing to evaluate if it is suitable for intoxication in ZEN contaminated animal feeds.

Zearalenone (ZEN), a Fusarium’s mycotoxin, is immunotoxic, genotoxic, hepatonephrotoxic and, affects the reproductive system. ZEN induces toxic and genotoxic effects on humans and other animals. Achyrocline satureioides has several medicinal properties. Moreover, the aqueous extract of A. satureioides is a safe agent that exerts low cytotoxicity and no genotoxicity. This extract is a promissory candidate to counteract ZEN effects. The present study aimed to investigate the capacity of cold aqueous extract from A. satureioides to protect against ZEN multi-target toxicity in different experimental mammal models. Anticytotoxicity was evaluated by neutral red uptake and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium reduction assays. Comet assay and micronuclei test, oxidative stress (TBARs), and histopathological damage were evaluated in Balb/C mice. Anticytotoxic studies indicated that cold aqueous extract (100 and 300 μg/ml) protected from damage induced by ZEN (50 μg/ml) on Vero cells. In vivo studies indicated that ZEN (40 mg/kg body weight) induced an increase of genotoxicity: micronuclei (34 MNPCE/1000 PCE) and increase of damage (tail moment) in blood cells. Also, it increased lipid peroxidation in liver and kidneys and generated several histopathological alterations in both organs. Cold aqueous extract (100 mg/kg body weight) protected from genotoxicity induced by ZEN in both tests. Cold aqueous extract, also, reduced the lipid peroxidation and histopathological damage in liver and kidneys. In conclusion, the cold aqueous extract of A. satureioides that contains bioactive flavonoids prevents the multi-target toxicity induced by ZEN improving all the parameters evaluated in vitro and in vivo, which is a valuable and original finding in order to develop future treatments for human and veterinary medicine.

The objective of this study was to evaluate the effects of the addition of Saccharomyces cerevisiae to feed contaminated with aflatoxin B1 (AFB1) on the performance and health of tambaqui fingerlings. A fully randomised study design was used, represented by four treatments: (T1) with 0.0 μg/kg of AFB1 and 0.0 colony forming unit (cfu)/g of yeast (control); (T2) with 0.0 μg/kg of AFB1 and 106 cfu/g of yeast; (T3) with 100 μg/kg of AFB1 and 0.0 cfu/g of yeast and (T4) with 100 μg/kg of AFB1 and 106 cfu/g of yeast, with four repetitions per treatment. Over a period of 34 days, 192 fingerlings with an initial weight of 2.9±0.2 g and length of 26.1±1.3 mm were used, distributed in 16 tanks with 12 fish per experimental unit. The following parameters were evaluated: water quality, zootechnical performance, histopathological analysis, counting and isolation of yeasts from the feed and intestine. No significant differences were observed (P>0.05) in the initial weight, initial length, final length, specific growth rate, mean length gain, Fulton condition factor (K Factor) or survival rate of the tambaqui fingerlings during the trial period. Statistical differences (P1. The S. cerevisiae A8L2 strain was able to colonise the intestine of tambaqui fingerlings, however, its addition in conjunction with 100 μg/kg AFB1 to fingerling feed did not promote improvements in performance indices or the histopathological parameters of these animals.

Latin America with its considerable North-South extent is subject to climate that varies from tropical, subtropical and warm temperate to temperate. Different agricultural products are produced in the area including cereals, oilseeds, beans, fruits and nuts together with animal production including cattle for beef and milk, pigs, poultry and fish. The heterogeneity of agriculture in Latin America is reflected in the diversity of the region's farm structures. While agriculture in the Southern Cone is dominated by large, commercial and export-oriented farms, particularly in Argentina and Brazil, besides increasingly in other countries like Uruguay, much of the rest of the region is characterised by smallholder and family agriculture. The contamination of agricultural products with mycotoxins has impact both human and animal health, as well as the economy due to losses related to rejections of agricultural products and by-products during trade. The economic burden related to the consumption of mycotoxins by animals is especially important, causing productivity losses up to the death of animals. The relevant mycotoxins are fumonisins, deoxynivalenol (DON) and zearalenone (ZEN) in cereals and cereal-based products, aflatoxins in cereals, oily seeds and nuts, aflatoxin M1 in milk and dairy products as well as ochratoxin A (OTA) in coffee, grapes and raisins. Co-occurrence of mycotoxins has also been observed mainly with aflatoxins and fumonisins in different Latin American countries (Torres et al., 2015). Advances on legislation in different countries including Argentina, Brazil, Chile, Mexico and Uruguay have been done to establish maximum limits for mycotoxins including aflatoxins, DON, ZEN, OTA, patulin and ergot alkaloids (ANVISA, 2011/2017; CAA, 2019/2021, Norma Oficial Mexicana, N.-243-S., 2010/2010; Reglamento Sanitario de los Alimentos, 2013). Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Torres, Adriana Mabel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Torres, Olga. Laboratorio Diagnóstico Molecular; Guatemala Fil: Mallmann, Carlos Alberto. Universidade Federal de Santa Maria; Brasil

The aim of the study was to determine whether the addition of vegetable biocholine (VB) in laying hens feed minimises the effects of daily intake of aflatoxin B1 (AFB1). We allocated Hy-line Brown line laying hens into four groups with four replications/group and four birds/repetition. The treatments were as follows: Afla0Bio0: basal feed without aflatoxin and VB (natural contamination: 0.026 mg AFB1/kg), Afla0Bio800, basal feed supplementation of 800 mg VB/kg (natural contamination: 0.024 mg AFB1/kg); Afla2.5Bio0, basal feed contaminated experimentally with aflatoxin (2.51 mg/kg); Afla2.5Bio800, basal feed contaminated with aflatoxin (2.50 mg/kg) and supplemented with 800 mg VB/kg. The experiment took place over a period of 42 days, divided into two cycles of 21 days each. Significance was indicated by P≤0.05. The inclusion of aflatoxin reduced egg production after 42 days of consumption of contaminated feed. VB supplementation in the tested dose was insufficient to minimise the negative effects of the toxin on the laying rate. There was a lower percentage of yolk in Afla2Bio0 than in Afla0Bio0, and a higher percentage of albumen and specific gravity in Afla2.5Bio0 than in Afla0Bio0. Ingestion of aflatoxin in the feed increased lipoperoxidation (LPO) and decreased antioxidant capacity in the egg yolk; however, when VB was added, LPO was similar to the control. Lower total bacterial count (TBC) in the eggshell was observed when the birds consumed VB, as well as higher TBC in the eggshell of the birds was challenged with aflatoxin. In the blood of birds that consumed aflatoxin (Afla2.5Bio0) there was an increase in the activity of alkaline phosphatase and a reduction in the activities of glutathione S-transferase and glutathione peroxidase (GPx). In the birds that consumed VB without aflatoxin challenge, we observed that there was a stimulation of GPx activity. We conclude that the consumption of VB had positive effects on the health of the laying hens and improved the quality of the eggs.