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SUMMARY The ability of naturally-occurring low levels of food-borne toxins and environmental contaminants to alter immune function in animals has been the subject of increasing interest in recent years. A total of 96 1-d-old broiler chickens were used to evaluate the effect of feeding naturally contaminated rations with low levels (800 ppb) of T-2 toxin and the protective effect of a commercial mycotoxin eliminator on circulating and intestinal immune cells, blood biochemistry, and hematological variables. Although there were no effects on body weight, contamination with T-2 toxin lead to a significant increase in serum levels of total protein (mainly due to increased albumin levels), alkaline phosphatase (AP), and aspartate aminotransferase (AST), and increased jejunal levels of CD3+ and Goblet cells at day 28. T-2 toxin significantly altered immune parameters, as was demonstrated by alterations in the number of circulating macrophages and suppressor macrophages, B-lymphocytes, T-lymphocytes from mucosa, virgin cytotoxic T-lymphocytes, and terminally activated cytotoxic T-lymphocytes compared to uncontaminated control birds. The use of a mycotoxin eliminator partly compensated for the changes in immune cells. The importance of low levels of mycotoxins is still debatable. However, in this research, it is clearly demonstrated that T-2 toxins affects immune responses, possibly due to impaired gut barrier functioning and metabolic changes.

A generic extraction and UHPLC-Q-Orbitrap™-HRMS method was developed for four insect species (mealworm, grasshopper, house cricket and black soldier fly) analyzing a large spectrum of organic chemical contaminants, including pesticides (n = 25), (veterinary) drugs (n = 29), and mycotoxins (n = 23). To prove the method as 'fit-for-purpose', a successful validation was performed, both qualitatively, by determining the screening detection limit (SDL), selectivity and specificity, as well as semi-quantitatively, by assessing the within-day precision (relative standard deviation (RSD)) and recovery. For both the mealworm, grasshopper, house cricket and black soldier fly, 64, 61, 59 and 62 compounds were detected at the respective SDL levels (1-100 μg kg-1), predominantly below existing maximum residue limits for other edible matrices. Mean recoveries ranged between 70% and 120% and RSD-values were in line with European regulations (CD 2002/657/EC; SANCO). Finally, the potential of the screening methodology was demonstrated on real insect samples, revealing minor to no contamination. ispartof: FOOD CHEMISTRY vol:293 pages:187-196 ispartof: location:England status: published

Co-occurrence of mycotoxins in food and feed represents the rule rather than the exception. Information about combinatory toxic effects of co-occurring mycotoxins is scarce, in particular the effects that mixtures of mycotoxins in feed may have on farm animals. This review focusses on studies on the combined effects of moniliformin and co-occurring mycotoxins in feed on farm animals. Moniliformin is a mycotoxin of emerging scientific interest, which may co-occur with many other mycotoxins, especially Fusarium mycotoxins. Oral exposure to moniliformin reduces feed consumption and body weight gain in poultry, in pigs and catfish, and induces cardiotoxic effects and/or alterations in serum biochemical and haematological parameters. In this review only experiments comparing effects as a result of the exposure to a combination of mycotoxins with effects due to the exposure to single mycotoxins were considered. Identified published studies on combined toxicity have been limited to combinations of moniliformin with either fumonisin B1 or deoxynivalenol, and were performed with poultry, pigs, and catfish. Most of the moniliformin/fumonisin B1 investigations involved poultry and focussed on adverse effects on feed intake, weight gain and immune response, as well as organ lesions. These studies mainly reported an interactive toxicity of moniliformin and fumonisin B1 but did not allow identification of the type of interaction. Likewise, no indication could be given for the interaction detected for both mycotoxins on weight gains of catfish. For the moniliformin/deoxynivalenol combination, only one study with broiler chickens was found relevant. This study concluded additive or less than additive toxicity, using kidney lesions and renal tubular epithelial degeneration as endpoints. While possible interactions between moniliformin and fumonisin B1 or deoxynivalenol were identified, the conclusions are based on limited studies and experimental designs. Further studies on the combined toxicity of moniliformin with other mycotoxins and other animal species would be needed.

The poultry industry in sub-Saharan Africa (SSA) is faced with feed insecurity, associated with high cost of feeds, and feed safety, associated with locally produced feeds often contaminated with mycotoxins. Mycotoxins, including aflatoxins (AFs), fumonisins (FBs), trichothecenes, and zearalenone (ZEN), are common contaminants of poultry feeds and feed ingredients from SSA. These mycotoxins cause deleterious effects on the health and productivity of chickens and can also be present in poultry food products, thereby posing a health hazard to human consumers of these products. This review summarizes studies of major mycotoxins in poultry feeds, feed ingredients, and poultry food products from SSA as well as aflatoxicosis outbreaks. Additionally reviewed are the worldwide regulation of mycotoxins in poultry feeds, the impact of major mycotoxins in the production of chickens, and the postharvest use of mycotoxin detoxifiers. In most studies, AFs are most commonly quantified, and levels above the European Union regulatory limits of 20 mu g/kg are reported. Trichothecenes, FBs, ZEN, and OTA are also reported but are less frequently analyzed. Co-occurrences of mycotoxins, especially AFs and FBs, are reported in some studies. The effects of AFs on chickens' health and productivity, carryover to their products, as well as use of mycotoxin binders are reported in few studies conducted in SSA. More research should therefore be conducted in SSA to evaluate occurrences, toxicological effects, and mitigation strategies to prevent the toxic effects of mycotoxins.

Mycotoxins are secondary metabolites of fungi that contaminate food and feed and have a significant negative impact on human and animal health and productivity. The tropical condition in Sub-Saharan Africa (SSA) together with poor storage of feed promotes fungal growth and subsequent mycotoxin production. Aflatoxins (AF) produced by Aspergillus species, fumonisins (FUM), zearalenone (ZEN), T-2 toxin (T-2), and deoxynivalenol (DON) produced by Fusarium species, and ochratoxin A (OTA) produced by Penicillium and Aspergillus species are well-known mycotoxins of agricultural importance. Consumption of feed contaminated with these toxins may cause mycotoxicoses in animals, characterized by a range of clinical signs depending on the toxin, and losses in the animal industry. In SSA, contamination of dairy feed with mycotoxins has been frequently reported, which poses a serious constraint to animal health and productivity, and is also a hazard to human health since some mycotoxins and their metabolites are excreted in milk, especially aflatoxin M1. This review describes the major mycotoxins, their occurrence, and impact in dairy cattle diets in SSA highlighting the problems related to animal health, productivity, and food safety and the up-to-date post-harvest mitigation strategies for the prevention and reduction of contamination of dairy feed.

Cytochrome P450 enzymes (CYP) are important catalyzing proteins involved in the biotransformation of endogenous and xenobiotic compounds. However, their expression and/or activity can be altered by exposure to contaminants such as mycotoxins. In vitro incubations in porcine hepatic microsomes revealed a potent inhibition of the midazolam (CYP3A) biotransformation by T-2 toxin (T-2) (Ki = 27.0 +/- 3.97 mu M) and zearalenone (ZEA) (Ki = 1.1 +/- 0.22 mu M). Consequently, the in vivo impact of 2 weeks exposure to T-2 (1,000 mu g/kg feed) or ZEA (500 mu g/kg feed) on the pharmacokinetics (PK) of midazolam (MDZ) as a CYP3A probe drug was investigated in pigs, and was compared to a control group receiving no mycotoxins. MDZ was chosen as this drug undergoes substantial first-pass metabolism in humans with equal contribution of the intestine and liver. Each pig received a single intravenous (0.036 mg/kg BW) and oral (0.15 mg/kg BW) dose of midazolam (MDZ). For the IV bolus no differences were observed in PK between control and mycotoxins exposed groups. However, oral plasma concentration-time profiles showed quantitative differences in absolute oral bioavailability F[p-value (ANOVA) = 0.022], AUC_0-inf (mu g*h/L) [p-value (ANOVA) = 0.023], Ke (1/h) [p-value (ANOVA) = 0.004], and Ka (1/h) [p-value (ANOVA) = 0.031]. Although only differences in Ke estimates after oral administration reached significance in the post hoc analysis due to inequality of the variances. We hypothesize that the observed trends after ZEA and T-2 exposure are related to the cytotoxic effect of T-2, resulting in an increased absorption rate constant Ka. For ZEA, an inhibition of the CYP3A enzymes is suggested based on the in vitro inhibition potential and increase in oral bioavailability. Further research is required to confirm the current hypothesis.

Forages are important components of dairy cattle rations but might harbor a plethora of mycotoxins. Ruminants are considered to be less susceptible to the adverse health effects of mycotoxins, mainly because the ruminal microflora degrades certain mycotoxins. Yet, impairment of the ruminal degradation capacity or high ruminal stability of toxins can entail that the intestinal epithelium is exposed to significant mycotoxin amounts. The aims of our study were to assess i) the mycotoxin occurrence in maize silage and ii) the cytotoxicity of relevant mycotoxins on bovine intestinal cells. In total, 158 maize silage samples were collected from European dairy cattle farms. LC-MS/MS-based analysis of 61 mycotoxins revealed the presence of emerging mycotoxins (e.g. emodin, culmorin, enniatin B1, enniatin B, and beauvericin) in more than 70% of samples. Among the regulated mycotoxins, deoxynivalenol and zearalenone were most frequently detected (67.7%). Overall, 87% of maize silages contained more than five mycotoxins. Using an in vitro model with calf small intestinal epithelial cells B, the cytotoxicity of deoxynivalenol, nivalenol, fumonisin B1 and enniatin B was evaluated (0&ndash, 200 &micro, M). Absolute IC50 values varied in dependence of employed assay and were 1.2&ndash, 3.6 &micro, M, 0.8&ndash, 1.0 &micro, M, 8.6&ndash, 18.3 &micro, M, and 4.0&ndash, 6.7 &micro, M for deoxynivalenol, nivalenol, fumonisin B1, and enniatin B, respectively. Results highlight the potential relevance of mycotoxins for bovine gut health, a previously neglected target in ruminants.

Physical methods, such as heating and irradiation with ultraviolet light (UVC), are among the strategies that may be used to reduce mycotoxin content in foodstuffs. This research studied the technological effects of heating at 100, 150, and 200 °C and UVC irradiation at 254 nm for 15, 30, 60, and 120 min applied on semolina to decrease the amount of mycotoxins. The impact of the most efficient treatments (150 °C for 30 min and UVC irradiation for 120 min) was evaluated both on semolina and dough technological properties, such as color, hydration, and gluten indices, farinograph and calorimetric parameters, and microstructure. Thermal treatment significantly increased hydration of semolina and swelling power; farinograph analyses revealed a marked increase in water absorption and dough development time and a reduction of dough stability and degree of softening and elasticity compared to the control. Calorimetric analyses showed a significant reduction in both temperature peaks and enthalpy of starch gelatinization and a decrease in enthalpy for melting of lipids with starch. Scanning electron micrograph images representing the dough microstructure confirmed the occurrence of these changes; UVC-treated samples showed less pronounced changes compared to the control. Practical applications: This work reports some strategies that could be integrated in semolina-based products chains, with the aim of reducing mycotoxin contamination. Thermal treatment and UVC irradiation are known to be cost-effective techniques, combining a high thermal efficiency with the simplicity of application. Understanding the technological changes resulting from these treatments could be useful to food companies for the realization of safer semolina-based products with different structural properties. This is the first study aiming to investigate the technological impacts of physical processing methods for reducing mycotoxin contamination.

International audience; Moniliformin (MON) is a mycotoxin with low molecular weight primarily produced by Fusarium fungi and occurring predominantly in cereal grains. Following a request of the European Commission, the CONTAM Panel assessed the risk of MON to human and animal health related to its presence in food and feed. The limited information available on toxicity and on toxicokinetics in experimental and farm animals indicated haematotoxicity and cardiotoxicity as major adverse health effects of MON. MON causes chromosome aberrations in vitro but no in vivo genotoxicity data and no carcinogenicity data were identified. Due to the limitations in the available toxicity data, human acute or chronic health-based guidance values (HBGV) could not be established. The margin of exposure (MOE) between the no-observed-adverse-effect level (NOAEL) of 6.0 mg/kg body weight (bw) for cardiotoxicity from a subacute study in rats and the acute upper bound (UB) dietary exposure estimates ranged between 4,000 and 73,000. The MOE between the lowest benchmark dose lower confidence limit (for a 5% response-BMDL 05) of 0.20 mg MON/kg bw per day for haematological hazards from a 28-day study in pigs and the chronic dietary human exposure estimates ranged between 370 and 5,000,000 for chronic dietary exposures. These MOEs indicate a low risk for human health but were associated with high uncertainty. The toxicity data available for poultry, pigs, and mink indicated a low or even negligible risk for these animals from exposure to MON in feed at the estimated exposure levels under current feeding practices. Assuming similar or lower sensitivity as for pigs, the CONTAM Panel considered a low or even negligible risk for the other animal species for which no toxicity data suitable for hazard characterisation were identified. Additional toxicity studies are needed and depending on their outcome, the collection of more occurrence data on MON in food and feed is recommended to enable a comprehensive human risk assessment.

Emerging Fusarium and Alternaria mycotoxins gain more and more interest due to their frequent contamination of food and feed, although in vivo toxicity and toxicokinetic data are limited. Whereas the Fusarium mycotoxins beauvericin, moniliformin and enniatins particularly contaminate grain and grain-based products, Alternaria mycotoxins are also detected in fruits, vegetables and wines. Although contamination levels are usually low (mu g/kg range), higher contamination levels of enniatins and tenuazonic acid may occasionally occur. In vitro studies suggest genotoxic effects of enniatins A, A1 and B1, beauvericin, moniliformin, alternariol, alternariol monomethyl ether, altertoxins and stemphyltoxin-III. Furthermore, in vitro studies suggest immunomodulating effects of most emerging toxins and a reproductive health hazard of alternariol, beauvericin and enniatin B. More in vivo toxicity data on the individual and combined effects of these contaminants on reproductive and immune system in both humans and animals is needed to update the risk evaluation by the European Food Safety Authority. Taking into account new occurrence data for tenuazonic acid, the complete oral bioavailability, the low total body clearance in pigs and broiler chickens and the limited toxicity data, a health risk cannot be completely excluded. Besides, some less known Alternaria toxins, especially the genotoxic altertoxins and stemphyltoxin III, should be incorporated in risk evaluation as well.

The aim of this trial was to assess the effects of the administration of different combinations of mycotoxins in naturally contaminated maize grains on dairy heifer growth, blood measurements and puberty onset. A total of 35 Friesian female heifers were randomly allotted to three experimental groups from 18-21 to 42-45 weeks of age. During the 24-week experimental period (EP), heifers were fed the same diet, but with maize meal derived from three differently contaminated lots: very low contamination, as control (C); medium-low aflatoxin-contaminated (A); and mixed aflatoxin-fumonisin contaminated (A-F). At the end of the EP, they returned to a common diet without contaminated maize, and they were monitored for an additional period of 12 weeks (post-experimental period, PEP). BW, wither height, hip height, body length and heart girth were measured every 4 weeks from the beginning of EP to the end of PEP. At the same time, body condition score was evaluated and blood samples were taken from the jugular vein to be analysed for haematological, serum protein and metabolic profiles. Age at puberty was assessed by measuring weekly plasma progesterone levels from 40 to 52 weeks of age. Body growth measurements were processed both by ANOVA of average daily gain of EP and PEP separately, and by the analysis of growth curve parameters. Haematological, serum protein and metabolic profile were evaluated using a mixed model, taking into account the repeated measurements in time on each animal. Heifers' growth was delayed both in A and A-F groups during EP, as evidenced by the different linear coefficients of the BW growth curve in the three groups. Differently contaminated diets did not affect the haematological profile, so that it can be concluded that these levels of mycotoxin contamination do not determine any specific effect on haematopoiesis and immunity in growing heifers. The main blood marker of mycotoxin chronic toxicity was the γ-glutamyl transferase activity level in plasma, which appeared to be altered even after the removal of mycotoxins. During EP, plasma glucose was lower in the groups fed contaminated diet compared with C. The joint actions of an altered nutritional status and a long-lasting liver damage were probably the causes of the delay in puberty attainment in A and, particularly, in the A-F group. The results from this trial evidenced that a chronic aflatoxin-fumonisin contamination in diets of dairy heifers can determine an important delay in the reproductive career of these animals.

This paper summarises workshop discussions at the 5th international MYCORED meeting in Ottawa, Canada (June 2012) with over 200 participants representing academics, government and industry scientists, government officials and farming organisations (present in roughly equal proportions) from 27 countries. Workshops centred on how mycotoxins in food and feed affect value chains and trade in the region covered by the North American Free Trade Agreement. Crops are contaminated by one or more of five important mycotoxins in parts of Canada and the United States every year, and when contaminated food and feed are consumed in amounts above tolerable limits, human and animal health are at risk. Economic loss from such contamination includes reduced crop yield, grain quality, animal productivity and loss of domestic and export markets. A systematic effort by grain producers, primary, transfer, and terminal elevators, millers and food and feed processers is required to manage these contaminants along the value chain. Workshops discussed lessons learned from investments in plant genetics, fungal genomics, toxicology, analytical and sampling science, management strategies along the food and feed value chains and methods to ameliorate the effects of toxins in grain on animal production and on reducing the impact of mycotoxins on population health in developing countries. These discussions were used to develop a set of priorities and recommendations.

Mycotoxins are secondary metabolites produced by fungi, which may be present on a variety of crops. They are considered a major issue worldwide because of their harmful effects on animals. These contaminants lead to great economic losses, especially in pig and poultry husbandry. Over 400 mycotoxins have been identifi ed. However, only few of them have a signifi cant toxic effect and are of major concern. In this paper, the most important mycotoxins are described, including deoxynivalenol (DON), T-2 toxin (T-2), zearalenone (ZON), fumonisin B1 (FB1), ochratoxin A (OTA) and afl atoxin B1 (AFB1). For each toxin, its chemical structure, mode of action and symptoms of acute and chronic toxicity in pigs and poultry are discussed.

This study was conducted to investigate mycotoxin exposure in children (n = 220, aged 1.5–4.5 years) from high mycotoxin contamination regions of Cameroon and to examine the association between the mycotoxin levels (in total 18 analytes) and several socio-demographic factors and anthropometric characteristics. A cross-sectional study was conducted in six villages in Cameroon with 220 children. Mycotoxins and their metabolites were detected in 160/220 (73%) urine samples.There were significant differences in the mean contamination levels of ochratoxin A (p = 0.01) and β-zearalenol (p = 0.017) between the two agro-ecological zones investigated. Likewise significant differences were observed in the mean levels of aflatoxin M1 (p = 0.001) across the weaning categories of these children. The mean concentration of aflatoxin M1 detected in the urine of the partially breastfed children (1.43 ng/mL) was significantly higher (p = 0.001) than those of the fully weaned children (0.282 ng/mL).Meanwhile, the mean concentrations of deoxynivalenol (3.0 ng/mL) and fumonisin B1 (0.59 ng/mL) detected in the urine of the male children was significantly (p value 0.021 for deoxynivalenol and 0.004 for fumonisin B1) different from the levels detected in the urine of female children; 0.71 ng/mL and 0.01 ng/mL for deoxynivalenol and fumonisin B1 respectively. In this study, there was no association between the different malnutrition categories (stunted, wasting and underweight) and the mycotoxin concentrations detected in the urine of these children.However, there is sufficient evidence to suggest that children in Cameroon under the age 5 are exposed to high levels of carcinogenic substances such as fumonisin B1, aflatoxin M1 and ochratoxin A through breastfeeding. To the best of our knowledge, this is the first report of its kind carried out in West Africa to determine multi-mycotoxin exposure in infants. Keywords: Mycotoxins, Biomarkers, Children, Urine, Weaned, Breastfeeding

BACKGROUND: Fumonisin B-1 (FB1) is a mycotoxin produced by several Fusarium species and is a very common contaminant of maize-based food and feed throughout the world. The selection and use of FB1-degrading microorganisms appears as a promising alternative to cope with the problem of toxicity towards humans and livestock. High moisture maize grain silage, which is based on natural maize fermentation, could be an interesting reservoir of such microorganisms. RESULTS: Using an in vitro simulated silage model with FB1 naturally contaminated grains, we demonstrated a significant raw decrease in FB1 during ensiling process ascribed to biodegradation mechanisms. A panel of 98 bacteria and yeasts were isolated from this matrix and selected for their ability to use FB1 as the sole source of C and N. For nine of them, the ability to degrade FB1 in vitro was evidenced. Notably, two bacteria identified as Lactobacillus sp. were highlighted for their efficient FB1-degrading capacity and production of hydrolysed FB1 as intermediate degradation metabolite. CONCLUSION: Fermentation of high moisture maize grain contaminated with FB1 leads to a significant reduction of the toxin and allows the isolation of FB1-degrading microorganisms that could further be used as FB1 decontaminating agents.

The present review summarises data on the prevalence and concentration of fusaproliferin, beauvericin and enniatins in small grains, maize and processed grain-based foods. These fungal secondary metabolites possess interesting biological activities and are presently often referred to as 'emerging mycotoxins'. They have been considered as less important since they are likely not of acute toxicity. However, their high prevalence in foodstuffs, occasionally in higher mg/kg concentrations, warrants an assessment of their true importance for food (and feed) safety. Thorough surveys that determine the contamination of raw and processed foods with fusaproliferin, beauvericin and enniatins are still scarce and are generally limited to Northern Europe and the Mediterranean. The reported data suggest a connection between the observed concentrations and climate. The data further suggest that humans might be exposed continuously to low levels of enniatins as these compounds were of particularly high prevalence in the surveys. The highest concentrations of enniatins were reported from Spain and Northern Africa (maximum concentration of enniatin A1 814 mg/kg in rice from the Spanish market), while concentrations were significantly lower in Northern Europe (maximum concentration of enniatin B 18.3 mg/kg in wheat from Finland). Beauvericin appears to be of low significance in grains from cooler climates while it has been reported to occur at concentrations of tens of mg/ kg in Southern Europe and Morocco (maximum concentration 59 mg/kg in maize from Morocco). Fusaproliferin has been least investigated. Several reports from the Mediterranean show its occasional occurrence up to 19.6 mg/ kg in rice from Morocco, while its natural occurrence in cooler climates seems to be more rare.

The natural occurrence of fumonisin B-1 (FB1) and its co-occurrence with zearalenone (ZEA), T-2 toxin and deoxynivalenol (DON) were surveyed in 103 winter wheat samples collected after four to six-month storage in family barns from different locations in Serbia. All 103 samples were mycotoxin positive. The mean concentrations of all mycotoxins except ZEA were greater in 2005 than in 2007. FB1 was detected in 82.1% and 92.0% of all samples with ranges of 750-5400 mu g kg(-1) (mean, 2079.45 mu g kg(-1)) and 750-4900 mu g kg(-1) (mean 918.76 mu g kg(-1)) in 2005 and 2007, respectively. Moderate positive correlations were found between FB1 and DON concentrations (r = 0.56 in 2005 and r = 0.54 in 2007) and between FB1 and ZEA concentrations (r = 0.48 in 2005 and r = 0.60 in 2007), while a moderate negative correlation was detected between the production of FB1 and T-2 toxin in 2007 (r = -0.33). This is the first report of FB1 occurrence in naturally-contaminated wheat grain and its simultaneous occurrence with ZEA, DON and T-2 toxin in Serbia. Moreover, this is one of the rare reports presenting the occurrence of FB1 on wheat in the world. (C) 2011 Elsevier Ltd. All rights reserved.

A wide variety of commodities in the world have been analyzed for fumonisins contamination. However, they have mostly been reported in maize and maize-based foods and feeds. Just a few scientific researches were conducted to obtain results on natural contamination of wheat and barley with these mycotoxins. This survey was conducted to evaluate fumonisin B1 contamination in maize, wheat and barley grain in Serbia. A total of 203 maize, 180 wheat and 120 barley samples were obtained from different local warehouses between October 2007 and June 2009. Concentration of FB1 were analysed with the Enzyme-Linked Immunosorbent Assay (ELISA). Positive results were found in 70.7%, 60.6% and 34.1%, in the maize, wheat and barley samples, respectively. FB 1 concentration varied from 750 to 4900 μg kg-1, and the mean levels recorded were: 1225.7 kg-1 in maize; 852.7 μg kg-1 in wheat; and 768.2 μg kg-1 in barley. The mycotoxin contamination of cereals was affected by factors such as origin resistance, droughtstress, and insect damage and differed between the years of investigation The results obtained in this survey revealed that FB1 is frequent contaminant of cereal grains in Serbia. Considering that these products are consumed in large amounts either directly or as components of foods and feeds, the levels of contamination reported herein indicate a potential threat to animal and public health. Širok spektar proizvoda u svetu je analiziran na kontaminaciju fumonizinima. Međutim, prisustvo ovog mikotoksina uglavnom je zabeleženo u zrnu kukuruza i hrani na bazi kukuruza. Malo je istraživanja o prirodnoj kontaminaciji pšenice i ječma fumonizinima. Cilj ovog istraživanja bio je da se ispita kontaminacija fumonizinom B1 zrna kukuruza, pšenice i ječma u Srbiji. Ukupan broj od 203 uzoraka zrna kukuruza, 180 uzoraka zrna pšenice i 120 uzoraka zrna ječma sakupljenih u različitim lokalitetima Srbije u periodu oktobar 2007. - jun 2009. godine. Koncentracija FB1 u zrnu žita je analizirana ELISA testom. FB1 je utvrđen u 72% uzoraka zrna kukuruza, 60,6% uzoraka zrna pšenice i 34,1% uzoraka zrna ječma. Koncentracija ovog mikotoksina je varirala od 750 do 4900 μg kg-1, sa srednjim vrednostima: 1225,7 kg-1 u kukuruzu; 852,7 μg kg-1 u pšenici; i 768,2 μg kg-1 u ječmu. Kontaminacija zrna sa FB1 je varirala u zavisnosti od mnogih faktora, kao što je otpornost biljke, stres suše, povrede od insekata i razlikovala se u ispitivanim godinama. Rezultati ovog istraživanja ukazali su da je FB1 značajan kontaminant zrna žita u Srbiji. S obzirom da se ovi proizvodi koriste u velikim količinama, bilo kao hrana za ljude, bilo kao hrana za životinje, utvrđeni stepen kontaminacije ukazuje na visok potencijalni rizik po javno zdravlje.

With an average annual production of 6.9 M tonnes and 2.5 M tonnes of maize and wheat respectively, Serbia is one of the main grain producers and exporters in Europe. Cereals are also the major staple food in Serbian diet. In view of the high cereal consumption, for human and animal nutrition, the presence of mycotoxins entails a high public health risk of chronic exposure to mycotoxins. This study provides an overview of the incidence of predominant mycotoxins, mainly in cereal and dairy products, in Serbia, in the 2004–2016, using data reported in the scientific literature. The study demonstrated that the total prevalence of aflatoxins was 62.9% (n = 12,517) with 26.2% of the samples exceeding the EU limits during this period. Results obtained for T-2/HT-2 (n = 523), deoxynivalenol (n = 2907), fumonisins (n = 998), zearalenone (n = 689) and ochratoxin A (n = 740) indicated the prevalence of 45.5%, 42.9%, 63.3%, 39.3% and 28.1%, respectively. For these mycotoxins, the EU limits were less frequently exceeded. Comprehensive collection and analysis of all accessible information reviewed in this paper showed moderate incidence and prevalence of mycotoxins in Serbia, with an exception of the 2012 drought year and the 2014 flood year.

High year-to-year variability in the incidence of Fusarium spp. and mycotoxin contamination was observed in a two-year survey investigating the impact of maize ear rot in 84 field samples from Germany. Fusarium verticillioides, F. graminearum, and F. proliferatum were the predominant species infecting maize kernels in 2006, whereas in 2007 the most frequently isolated species were F. graminearum, F. cerealis and F. subglutinans. Fourteen Fusariumrelated mycotoxins were detected as contaminants of maize kernels analyzed by a multi-mycotoxin determination method. In 2006, a growth season characterized by high temperature and low rainfall during anthesis and early grain filling, 75% of the maize samples were contaminated with deoxynivalenol, 34% with fumonisins and 27% with zearalenone. In 2007, characterized by moderate temperatures and frequent rainfall during the entire growth season, none of the 40 maize samples had quantifiable levels of fumonisins while deoxynivalenol and zearalenone were detected in 90% and 93% of the fields, respectively. In addition, 3-acetyldeoxynivalenol, 15-acetyldeoxnivalenol, moniliformin, beauvericin, nivalenol and enniatin B were detected as common contaminants produced in both growing seasons. The results demonstrate a significant mycotoxin contamination associated with maize ear rots in Germany and indicate, with regard to anticipated climate change, that fumonisins-producing species already present in German maize production may become more important. Keywords Deoxynivalenol . Ear rot . F. verticillioides . F. graminearum . Fumonisin . Zearalenone

In a survey carried out on 87 rotted fig fruits samples collected in the Apulia region of Italy, the authors isolated 126 Fusarium strains identified as F. ramigenum (69 strains), F. solani (49), F. proliferatum (five) and three not identified. Investigation on the fertility of the strains belonging to F. proliferatum and F. ramigenum revealed that only strains of F. proliferatum were fertile. The identity of F. ramigenum strains was confirmed by sequencing a portion of the translation elongation factor-1alpha gene. When Fusarium species were analysed for their toxigenicity, 37/69 strains of F. ramigenum produced fusaric acid (FA) up to 525 mg kg(-1); 30 strains produced beauvericin (BEA) up to 190 mg kg(-1); 60 strains produced fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)) up to 1575 mg kg(-1) of total FBs; and two strains produced fusaproliferin (FUP) up to 345 mg kg(-1); all five strains of F. proliferatum produced FA at low levels; two strains produced BEA up to 205 mg kg(-1); one strain produced FB(1) and FB(2), 1100 and 470 mg kg(-1), respectively; and one strain produced FUP, 820 mg kg(-1); F. solani (30 strains) produced FA, 13 strains up to 215 mg kg(-1). Few fungal extracts showed high toxicity toward brine shrimp larvae and in some cases in relation to BEA and FA content. A pathogenic assay on fig fruits showed that all three species were pathogenic, with higher virulence of F. ramigenum. These data report for the first time the production of BEA and FB(1)/FB(2) by F. ramigenum and show that it is a main agent of fig endosepsis in Apulia and can contribute to fumonisin contamination of fresh and dried figs.

Fusarium proliferatum can occur on a wide range of economically important vegetable plants but its role in disease is not always well established. In 2000 and 2001, from forty-one field samples of wilting onion and garlic plants in Serbia, F. proliferatum as the predominant fungal species was isolated from root and bulbs. Seventy isolates were firstly characterized for their sexual fertility and were shown to be mostly members of Gibberella intermedia (sixty-seven of seventy isolates, the remaining three isolates were unfertile), the sexual stage of F. proliferatum (syn. mating population D of G. fujikuroi complex). A selected set of eleven F. proliferatum isolates from both hosts were also tested for their pathogenicity and toxigenicity. Although onion and garlic plants were susceptible to all isolates, onion plants showed a significantly higher disease severity index. Six of the eleven isolates of F. proliferatum produced fumonisin B-1 from 25 to 3000 mu g g(-1), and beauvericin from 400 to 550 mu g g(-1); ten isolates produced fusaric acid from 80 to 950 mu g g(-1) and moniliformin from 50 to 520 mu g g(-1). Finally, all isolates produced fusaproliferin up to 400 mu g g(-1). These results confirm F. proliferatum as an important pathogen of garlic and onion in Europe and that there is a potential mycotoxin accumulation risk in contaminated plants of both garlic and onion.

Mycotoxins, toxic secondary metabolites of fungi, affect global agriculture so prolifically that they are virtually ubiquitous at some concentration in the average human diet. Studies of in vitro and in vivo toxicity are discussed, leading to investigations of co-exposed mycotoxins, as well as carcinogenic effects. Some of the most common and toxicologically significant mycotoxins, such as the aflatoxins, ochratoxins, fumonisins, deoxynivalenol, T-2 toxin, HT-2 toxin, patulin, zearalenone, and some ergot alkaloids are outlined. The wide variety of pathogenic mechanisms these compounds employ are shown capable of inducing a complex set of interactions. Of particular note are potential synergisms between mycotoxins with regard to carcinogenic attributable risk, indicating an important field for future study.

The objective of this study was to evaluate the occurrence and levels of deoxynivalenol (DON), fumonisins B-1 and B-2 (FBs), and zearalenone (ZEN) contaminants in animal feeds used in Korea in 2012. Contamination with DON was observed in 91.33% and 53.33% in compound feeds and feed ingredients, respectively. Among compound feeds, poultry layer feed (laying) exhibited the highest contaminant level of 1.492 mg/kg. FBs contaminants were present in compound feeds and feed ingredients at 93.33% and 83.33%, respectively. Most poultry broiler (early) feeds were highly contaminated with FBs, and one of these feeds detected the level as 12.823 mg/kg as the highest level. The levels of ZEN in compound feeds and feed ingredients were 71.33% and 47%, respectively. Ninety-eight percent of compound feeds for cattle were contaminated with ZEN, and the highest contamination level of 0.405 mg/kg was observed in cattle fatting feeds.

Contamination of food and feed with mycotoxins is a worldwide problem. At present, acute mycotoxicosis caused by high doses is rare in humans and animals. Ingestion of low to moderate amounts of Fusarium mycotoxins is common and generally does not result in obvious intoxication. However, these low amounts may impair intestinal health, immune function and/or pathogen fitness, resulting in altered host pathogen interactions and thus a different outcome of infection. This review summarizes the current state of knowledge about the impact of Fusarium mycotoxin exposure on human and animal host susceptibility to infectious diseases. On the one hand, exposure to deoxynivalenol and other Fusarium mycotoxins generally exacerbates infections with parasites, bacteria and viruses across a wide range of animal host species. Well-known examples include coccidiosis in poultry, salmonellosis in pigs and mice, colibacillosis in pigs, necrotic enteritis in poultry, enteric septicemia of catfish, swine respiratory disease, aspergillosis in poultry and rabbits, reovirus infection in mice and Porcine Reproductive and Respiratory Syndrome Virus infection in pigs. However, on the other hand, T-2 toxin has been shown to markedly decrease the colonization capacity of Salmonella in the pig intestine. Although the impact of the exposure of humans to Fusarium toxins on infectious diseases is less well known, extrapolation from animal models suggests possible exacerbation of, for instance, colibacillosis and salmonellosis in humans, as well.

The aim of this manuscript was the development of easy-to-operate quantum dots (QDs)-based immunochemical techniques for simultaneous screening of several mycotoxins in cereals. Two different approaches for multiplex fluorescent immunosorbent assay (FLISA) were employed. In the first approach a multiwell plate in which the different wells express a different mycotoxin (deoxynivalenol, zearalenone, aflatoxin B1, T2-toxin and fumonisin B1) was considered as a multiplex because each sample was pretreated once and then will be distributed over a series of wells within the same plate (single-analyte multiplex, SAM). The entire assay allows the simultaneous determination of all compounds. For the double-analyte multiplex (DAM) two different specific antibodies were co-immobilized in one single well. Zearalenone and aflatoxin B1 were simultaneously determined, provided their conjugates are labeled with QDs which are fluorescent in different parts of the spectrum, by scanning the assay outcome at two different wavelengths. The limits of detection (LOD) for the simultaneous determination of deoxynivalenol, zearalenone, aflatoxin B1, T2-toxin and fumonisin B1 by SAM FLISA were 3.2, 0.6, 0.2, 10 and 0.4 µg kg(-1), respectively, while for the DAM FLISA they were 1.8 and 1 µg kg(-1) for zearalenone and aflatoxin B1, respectively. SAM FLISA principle was also presented in a qualitative on-site format and tested for on-site multiplex determination of four mycotoxins in cereals. The achieved cut-off values of 500, 100, 2 and 100 µg kg(-1) for deoxynivalenol, zearalenone, aflatoxin B1 and T2-toxin respectively. For simplification of multiassay results' evaluation the conjugates with QDs of different colors were used.

Fusarium verticillioides is one of the most important fungal pathogens causing ear and stalk rot in maize. Even if frequently asymptomatic, it can produce a harmful series of compounds named fumonisins. Plant and fungal oxylipins play a crucial role in determining the outcome of the interaction between the pathogen and its host. Moreover, oxylipins are factors able to modulate the secondary metabolism in fungi. To uncover the existence of the relationship between oxylipin production and fumonisin synthesis in F. verticillioides, we analysed some molecular and physiological parameters, such as the expression of genes whose products are related to oxylipin synthesis (i.e. lipoxygenase, diol synthases and fatty acid oxidase), the oxylipin profile of both cracked maize and the pathogen by using a lipidomic approach (i.e. combining LC-TOF and LC-MS/MS approaches with a robust statistical analysis) and the synthesis of fumonisin B1. The results suggested a close relationship between the modification of the pathogen oxylipin profile with the fumonisin synthesis. Notably, a modification of the oxylipin profile of the pathogen during its growth on cracked maize can be demonstrated. The switch in oxylipin synthesis could indicate that the ‘presence’ of maize determinants (e.g. plant cell wall fragments and/or lipids) was able to promote the modification of the pathogen lifestyle, also by adapting the secondary metabolism, notably fumonisin synthesis.

In Nigeria, maize, sorghum, and millet are very important cash crops. They are consumed on a daily basis in different processed forms in diverse cultural backgrounds. These crops are prone to fungi infestation, and subsequently may be contaminated with mycotoxins. A total of 363 samples comprising of maize (136), sorghum (110), millet (87), and ogi (30) were collected from randomly selected markets in four agro-ecological zones in Nigeria. Samples were assessed for Fusarium mycotoxins contamination using a multi-mycotoxin liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Subsequently, some selected samples were analysed for the occurrence of hidden fumonisins. Overall, 64% of the samples were contaminated with at least one toxin, at the rate of 77%, 44%, 59%, and 97% for maize, sorghum, millet, and ogi, respectively. Fumonisins were the most dominant, especially in maize and ogi, occurring at the rate of 65% and 93% with mean values of 935 and 1128 mu g/kg, respectively. The prevalence of diacetoxyscirpenol was observed in maize (13%), sorghum (18%), and millet (29%), irrespective of the agro-ecological zone. Other mycotoxins detected were deoxynivalenol, zearalenone, and their metabolites, nivalenol, fusarenon-X, HT-2 toxin, and hidden fumonisins. About 43% of the samples were contaminated with more than one toxin. This study suggests that consumption of cereals and cereal-based products, ogi particularly by infants may be a source of exposure to Fusarium mycotoxins.

Background The gastrointestinal tract is the first target for the potentially harmful effects of mycotoxins after intake of mycotoxin contaminated food or feed. With deoxynivalenol (DON), T-2 toxin (T-2), fumonisin B1 (FB1) and zearalenone (ZEA) being important Fusarium toxins in the northern hemisphere, this study aimed to investigate in vitro the toxic effect of these mycotoxins on intestinal porcine epithelial cells derived from the jejunum (IPEC-J2 cells). Viability of IPEC-J2 cells as well as the proportion of apoptotic and necrotic IPEC-J2 cells was determined by flow cytometry after 72 h of exposure to the toxins. Correlatively, the integrity of the intestinal epithelial cell monolayer was studied using Transwell® inserts, in which the trans-epithelial electrical resistance (TEER) and passage of the antibiotics doxycycline and paromomycin were used as endpoints. Results We demonstrated that the percentage of Annexin-V-FITC and PI negative (viable) cells, Annexin-V-FITC positive and PI negative (apoptotic) cells and Annexin-V-FITC and PI positive (necrotic) IPEC-J2 cells showed a mycotoxin concentration-dependent relationship with T-2 toxin being the most toxic. Moreover, the ratio between Annexin-V-FITC positive and PI negative cells and Annexin-V-FITC and PI positive cells varied depending on the type of toxin. More Annexin-V-FITC and PI positive cells could be found after treatment with T-2 toxin, while more Annexin-V-FITC positive and PI negative cells were found after exposure to DON. Consistent with the cytotoxicity results, both DON and T-2 decreased TEER and increased cellular permeability to doxycycline and paromomycin in a time- and concentration-dependent manner. Conclusions It was concluded that Fusarium mycotoxins may severely disturb the intestinal epithelial barrier and promote passage of antibiotics.

The predominant species in maize in temperate climates is Fusarium graminearum, which produces the mycotoxins deoxynivalenol and zearalenone. Projected climate change is expected to affect Fusarium incidence and thus the occurrence of these mycotoxins. Predictive models may be helpful in determining trends in the levels of these mycotoxins with expected changing climatic conditions. The aim of this study was to develop a model describing fungal infection and subsequent growth as well as the formation of deoxynivalenol and zearalenone in maize in The Netherlands. For this purpose, a published Italian model was used as a starting point. This model is a mixed empiric–mechanistic model that describes fungal infection during silking (based on wind speed and rainfall) and subsequent germination, growth and toxin formation (depending on temperature and water availability). Model input uses weather parameters and crop management factors, such as maize hybrid, sowing date, flowering period and harvest date. Model parameter values were obtained by fitting these parameters to deoxynivalenol and zearalenone measurements in Dutch maize, using national mycotoxin data from the years 2002–2007. The results showed that the adapted model is capable of describing the trend in average deoxynivalenol and zearalenone levels over these years. Validation with external data is needed to verify model outcomes. It is expected that the current model can be used to estimate the effect of projected climate change on trends in deoxynivalenol and zearalenone levels in the coming years.

Two multi-analyte flow-through immunoassay formats for rapid detection of mycotoxins in a variety of food matrices (peanut cake, maize, and cassava flour) were developed and evaluated. The selected food matrices are typical staple foods and export products for most low-income communities around the world. The assay formats included gel-based and membrane-based flow-through assays and were based on the principle of indirect enzyme-linked immunosorbent assay. Using the same immunoreagents, the performance characteristics of both assays were compared. To the best of our knowledge, this is the first report on such a comparison. The gel-based format was developed to screen for ochratoxin A, fumonisin B(1), deoxynivalenol, and zearalenone detection at cut-off values of 3, 1,250, 1,000, and 200 μg kg(-1), respectively, while the membrane-based format can be used to screen ochratoxin A, aflatoxin B(1,) deoxynivalenol, and zearalenone at the following cut-offs: 3, 5, 700, and 175 μg kg(-1), respectively. The applicability of these assay formats was demonstrated by evaluating the performance characteristics of both tests through performing multiple experiments on different days. Both assays were further evaluated by analyzing naturally contaminated samples in the laboratory and also in the field under tropical conditions (Cameroon, West Africa). The false-negative rate with both formats was less than 5%, which is in good agreement with Commission Decision 2002/657/EC regarding the performance of analytical methods intended for screening purposes.

Samples of wheat grain (41), collected during the 2010 harvest from seven localities in Serbia, were analyzed for the presence of zearalenone (ZEA), T-2 toxin, deoxynivalenol (DON) and fumonisine B1 (FB1). Results of Enzyme-Linked Immunosorbent Assay (ELISA) showed that all analysed samples were positive for the presence of at least one of four observed fusariotoxins. The most distributed mycotoxins were ZEA (90.2%, with the average concentration of 442.6μg kg-1) and T-2 (90.2%, with the average concentration of 24.2 μg kg-1). DON (73.2%) and FB1 (84.4%) were detected in a somewhat smaller number of samples, but their average concentrations were higher (1988.1 μg DON kg-1 and 882.7 μg FB1 kg-1). The established correlations between concentrations of DON and FB1 (r = 0.32) or DON and ZEA (r = 0.22) were not statistically significant. A negative correlation was established between concentrations of T-2 and FB1 (r= -0.24), as well as, between T-2 and DON (r = -0.36). Detected concentrations of ZEA and T-2 were bellow the level prescribed by the World Health Organisation (WHO), while concentrations of FB1 and DON detected in five that is, 17 samples, respectively, were above the permissible limit for human consumption. Uzorci zrna pšenice (41), sakupljeni tokom žetve 2010. godine iz 7 različitih lokaliteta na teritoriji Srbije, analizirani su na prisustvo ZEA, T-2 toksina, deoksinivalenola (DON) i FB1. Primenom ELISA metode utvrđeno je da su svi analizirani uzorci bili pozitivni na prisustvo bar jednog od četiri ispitivana fuzariotoksina. Najzastupljeniji mikotoksini su bili ZEA (90,2%, prosečne koncentracije 442,6 μg kg-1) i T-2 (90,2%, prosečna koncentracija 24,2 kg-1). U nešto manjem broju uzoraka utvrđeni su DON (73,2%) i FB1 (84,4%), ali u većoj prosečnoj koncentraciji (1988,1 μg DON kg-1 i 882,7 μg FB1 kg-1). Nije utvrđena statistički značajna korelacija između koncentracije DON i FB1 (r = 0,32) ili DON i ZEA (r = 0,22). Negativna korelacija je utvrđena između koncentracije T-2 i FB1 (r = -0,24), kao i između T-2 i DON (r = -0,36). Detektovane koncentracije ZEA i T-2 bile su niže od nivoa koji propisuje Svetska zdravstvena organizacija, dok su FB1 i DON detektovani kod 5, odnosno 17 uzoraka u koncentraciji većoj od dozvoljenih u ljudskoj hrani.

Natural Zeolitic tuff was modified with 2, 5 and 10 mmol M+/100 g of octadecyldimethylbenzyl ammonium (ODMBA) ions and the products were denoted as OZ-2, OZ-5 and OZ-10. The starting material and organozeolites were characterized by determination of the point of zero charge (pH(pzc)) and by thermal analysis. in vitro sorption of fumonisin B-1 (FB1) was studied for all sorbents at pH 3, 7 and 9. The pH(pzc) for the zeolitic tuff was 6.8 +/- 0.1, while the pH(pzc) for OZ-2, OZ-5 and OZ-10 pH(pzc) was 7.0 +/- 0.1. The curves pH(final) = f(pH(initial)) suggest that the surfaces of all sorbents are positively charged at pH 3 and uncharged at pH 7 and 9. High sorption of FB1 by the zeolitic tuff in acidic solution Suggests electrostatic Interactions between the anionic FB1 and the positively charged surface. At pH 7 and 9, adsorption of FB1 is prevented because anionic FB1 cannot be adsorbed at the uncharged surface. From the pH(pzc) for the organozeolites, it is possible that with lower amounts of ODMBA (OZ-2 and OZ-5), at pH 3, beside interactions between head groups of ODMBA and its alkyl chains and anionic FB1, electrostatic interactions between positive uncovered surface and anionic FB1 contribute to the sorption, while at pH 7 and 9 there is only the possibility for interactions between FB1 and ODMBA. When the zeolitic surface was completely covered with ODMBA (OZ-10), FB1 sorption was independent of the form of FB1 suggesting only interactions between ODMBA and FB1. (C) 2009 Elsevier B.V. All rights reserved.

A method is described using LC-MS for the detection of the mycotoxins fusaproliferin (FUS) and beauvericin (BEA) in cultures of Fusarium subglutinans and in naturally contaminated maize. Protonated molecular ion signals for FUS and BEA were observed at m/z 445 and m/z 784, respectively. Collision induced dissociation of the readily dehydrated protonated molecular ion of the sesterterpene FUS (m/z 427) led to the loss of another water molecule (m/z 409) and acetic acid (m/z 385), while the cyclic lactone trimer BEA fragmented to yield the protonated dimer (m/z 523) and monomer (m/z 262), respectively. Detection of FUS was best performed in the MS-MS mode while BEA displayed a stronger signal in the MS mode. The on-column instrumental detection limits for pure FUS and BEA were found to be 2 ng and 20 pg (S/N = 2) while those in naturally contaminated maize were 1 mu g/kg and 0.5 mu g/kg, respectively. Five South African strains of F. subglutinans were analyzed following methanol extraction of which four produced FUS at levels between 330 mg/kg and 2630 mg/kg while only three produced BEA at levels between 140 mg/kg and 700 mg/kg. Application of this method to naturally contaminated maize samples from the Transkei region of South Africa showed FUS at levels of 8.8-39.6 mu g/kg and BEA at 7.6-238.8 mu g/kg. (C) 1999 Elsevier Science B.V. All rights reserved.

Toxicological profile and fertility of twenty four isolates of P. proliferatum, isolated from maize plants grown in different localities across Serbia, were analyzed. Seven investigated isolates were MATD-1 and seventeen were MATD-2. All isolates produced FB1 at a relatively high level (up to 3950 mu g g(1)), except for MRIZP56 which did not produce any FB1. Three isolates did not produce BEA and two just in traces. All isolates produced FUP, although three in traces and two at a relatively low level (50 and 150 mu g g(-1)). Mycotoxins production distinguish between isolates originated from different part of maize plants.