Moniliformin in cereal grains available in the Japanese market

ABSTRACTThe aim of the project was to produce updated information during 2005–14 on the Fusarium species found in Finnish cereal grains, and the toxins produced by them, as the last comprehensive survey study of Fusarium species and their toxins in Finland was carried out at the turn of the 1960s and the 1970s. Another aim was to use the latest molecular and chemical methods to investigate the occurrence and correlation of Fusarium species and their mycotoxins in Finland. The most common Fusarium species found in Finland in the FinMyco project 2005 and 2006 were F. avenaceum, F. culmorum, F. graminearum, F. poae, F. sporotrichioides and F. langsethiae. F. avenaceum was the most dominant species in barley, spring wheat and oat samples. The occurrence of F. culmorum and F. graminearum was high in oats and barley. Infection by Fusarium fungi was the lowest in winter cereal grains. The incidence of Fusarium species in 2005 was much higher than in 2006 due to weather conditions. F. langsethiae has become much more common in Finland since 2001. F. graminearum has also risen in the order of importance. A highly significant correlation was found between Fusarium graminearum DNA and deoxynivalenol (DON) levels in Finnish oats, barley and wheat. When comparing the FinMyco data in 2005–06 with the results of the Finnish safety monitoring programme for 2005–14, spring cereals were noted as being more susceptible to infection by Fusarium fungi and the formation of toxins. The contents of T-2 and HT-2 toxins and the frequency of exceptionally high DON concentrations all increased in Finland during 2005–14. Beauvericin (BEA), enniatins (ENNs) and moniliformin (MON) were also very common contaminants of Finnish grains in 2005–06. Climate change is leading to warmer weather, and this may indicate more changes in Finnish Fusarium mycobiota and toxin contents and profiles in the near future.

<em>Fusarium</em>-ear blight is a destructive disease in various cereal-growing regions and leads to significant yield and quality losses for farmers and to contamination of cereal grains with mycotoxins, mainly deoxynivalenol and derivatives, zearalenone and moniliformin. <em>Fusarium</em> pathogens grow well and produce significant inoculum on crop resiudues. Reduction of mycotoxins production and pathogen sporulation may be influenced by saprophytic fungi, exhibiting antagonistic effect. Dual culture bioassays were used to examine the impact of 92 isolates (belonging to 29 fungal species) against three toxigenic species, i.e. <em>Fusarium avenaceum</em> (Corda) Saccardo, <em>F. culmorum</em> (W.G.Smith) Saccardo and <em>F. graminearum</em> Schwabe. Both <em>F.culmorum</em> and <em>F. graminearum</em> isolates produce trichothecene mycotoxins and mycohormone zearalenone and are considered to be the most important cereal pathogens worldwide. Infection with those pathogens leads to accumulation of mycotoxins: deoxynivalenol (DON) and zearalenone (ZEA) in grains. <em>Fusarium avenaceum</em> isolates are producers of moniliformin (MON) and enniatins. Isolates of <em>Trichoderma</em> sp. were found to be the most effective ones to control the growth of examined <em>Fusarium</em> species. The response of <em>Fusarium</em> isolates to antagonistic activity of <em>Trichoderma</em> isolates varied and also the isolates of <em>Trichoderma</em> differed in their antagonistic activity against <em>Fusarium</em> isolates. The production of MON by two isolates of F. avenaceum in dual culture on rice was reduced by 95% to 100% by <em>T. atroviride</em> isolate AN 35. The same antagonist reduced the amount of moniliformin from 100 μg/g to 6.5 μg/g when inoculated to rice culture contaminated with MON, which suggests the possible decomposition of this mycotoxin.

Mycotoxin contamination in agricultural goods is a major global problem due to its negative impact on human and animal health. The principal mycotoxin producers are fungal species from the genera Fusarium, Aspergillus, Alternaria, and Penicillium. The toxigenic fungal species produce the mycotoxins as secondary metabolites when they invade agricultural commodities during crop cultivation in the field (preharvest) or after harvesting or during transport and storage. This study was designed to investigate the levels of Fusarium mycotoxins, viz., fumonisin (FUM), zearalenone (ZEN), and deoxynivalenol (DON) in cereal grain samples collected from Muscat, Sultanate of Oman during 2023-24. A total of 90 cereal grain (wheat, corn, rice, barley) samples from local markets at Muscat, the Plant Quarantine Department, Oman, and Oman Flour Mills Company were analyzed using competitive enzyme immunoassay kits. Furthermore, Fusarium spp. associated with the contaminated grain samples were isolated, and their mycotoxin-producing potential was assessed. The results indicated that FUM, ZEN, and DON levels were below the detection limit (LOD) in 81%, 97%, and 44% of the samples, respectively. Two out of fifteen corn samples and one out of thirty-seven wheat samples tested exceeded the maximum permissible limit for FUM and ZEN, respectively, as set by the European Commission. A total of 19 Fusarium spp. associated with the contaminated grain samples were isolated and identified through molecular techniques. Sixteen isolates of F. verticillioides, one isolate of F. thapsinum, and two new Fusarium species were identified based on nuclear ribosomal DNA internal transcribed spacer and elongation factor 1-alpha sequences. Two isolates of F. verticillioides (FQD-1 and FQD-20) produced FUM levels exceeding 2000 µg kg−1. The maximum ZEN concentration was observed in F. verticillioides FQD-20 (9.2 µg kg−1), followed by F. verticillioides FQD-2 (2.8 µg kg−1) and Fusarium sp. FOFMC-26 (2.5 µg kg−1). All tested Fusarium strains produced DON, with levels ranging from 25.6 to 213 µg kg−1, with F. thapsinum FQD-4 producing the highest level (213 µg kg−1). To our knowledge, this is the first report on the occurrence of Fusarium mycotoxins and mycotoxigenic Fusarium spp. in food commodities in Oman.

Five species of Fusarium, F. proliferatum, F. fujikuroi, F. verticillioides, F. sacchari, and Fusarium sp. are associated with pineapple fruit rot and leaf spot diseases in Malaysia. Their ability to produce mycotoxins was analysed by the detection of the FUM1 gene as an indicator of fumonisin B1 (FB1) production and by quantification of FB1, moniliformin (MON), and beauvericin (BEA) using ultra performance liquid chromatography. All isolates of F. proliferatum, F. fujikuroi, and F. verticillioides showed the presence of FUM1 band and were able to produce FB1 in vitro. MON was produced by all isolates of Fusarium species, however, BEA was produced only by F. verticillioides isolates and some F. proliferatum, F. fujikuroi, and Fusarium sp. isolates. Production of FB1, MON, and BEA in vitro can be used to predict mycotoxigenic potential of the Fusarium spp. associated with pineapple diseases.

A straightforward analytical method was developed and validated to determine the mycotoxin moniliformin in cereal-based foods. Moniliformin is extracted with water and quantified with liquid chromatography tandem mass spectrometry, and its presence confirmed with liquid chromatography-Orbitrap-high-resolution mass spectrometry. The method was validated for flour, bread, pasta and maize samples in terms of linearity, matrix effect, recovery, repeatability and limit of quantification. Quantification was conducted by matrix-matched calibration. Positive samples were confirmed by standard addition. Recovery ranged from 77 to 114% and repeatability from 1 to 14%. The limit of quantification, defined as the lowest concentration tested at which the validation criteria of recovery and repeatability were fulfilled, was 10 μg/kg. The method was applied to 102 cereal-based food samples collected in the Netherlands and Germany. Moniliformin was not detected in bread samples. One of 22 flour samples contained moniliformin at 10.6 μg/kg. Moniliformin occurred in seven out of 25 pasta samples at levels around 10 μg/kg. Moniliformin (MON) was present in eight out of 23 maize products at levels ranging from 12 to 207 μg/kg.

A survey on Fusarium species and moniliformin (MON) occurrence in sorghum grains collected from one of the main sorghum-producing areas of Argentina was conducted. Also, growth of F. thapsinum, one of the main sorghum pathogens, and MON production under different water activity (aw ) conditions on a sorghum-based medium were determined. Infection of sorghum grains by Fusarium species ranged from 82.5 to 99%; closely related species F. verticillioides, F. thapsinum and F. andiyazi were the most frequently recovered, followed by F. proliferatum and F. subglutinans. By sequencing a portion of the translation elongation factor-1α (TEF-1α) gene and by maximum parsimony analysis, F. verticillioides and closely related species were identified as F. thapsinum, F. andiyazi and F. verticillioides. Species within the F. graminearum species complex (FGSC) were isolated in high frequency. Maximum growth rates of 12 F. thapsinum strains were obtained at 0.995 aw . All evaluated strains were able to produce MON at all aw values tested, but MON production was higher at 0.995-0.982 aw . MON was detected in 41% of the samples at levels ranging from 363.2 to 914.2 µg kg-1 . This study provides new data on the occurrence of Fusarium species in sorghum grains destined for animal consumption in Argentina. The production of MON at different aw values showed that the toxin can be produced under field conditions. The risk to livestock exposed to daily low levels of MON associated with the toxin occurrence in the sorghum grains analyzed is unknown. © 2018 Society of Chemical Industry.

Deoxynivalenol and other selected Fusarium toxins in Swedish oats — Occurrence and correlation to specific Fusarium species

The European corn borer (ECB), Ostrinia nubilalis Hb., is a major pest of maize in Central Europe and is suspected to promote infection of maize with Fusarium species. The objectives of this study were to (1) determine moniliformin (MON) concentration in early maturing European Bt maize hybrids, their isogenic counterparts, commercial cultivars and experimental hybrids, (2) evaluate the association between MON concentration and ECB resistance and (3) correlate MON concentration with concentrations of other mycotoxins determined from the same plant materials. The field experiments were performed at five locations in Germany. MON concentration was significantly higher with manual infestation of ECB larvae (296 μg/kg) than under insecticide protection (66 μg/kg). Bt hybrids showed significantly lower MON concentrations and higher grain yields under manual ECB infestation than their corresponding isogenic counterparts, as well as commercial and experimental hybrids. All ECB resistance traits and grain yield under ECB infestation were significantly correlated with MON concentration. Correlations between concentrations of MON and other Fusarium mycotoxins were not significant. The use of Bt maize hybrids or insecticides to control ECB reduces the contamination of maize grains with MON in Central Europe. The presence of resistance genes against Fusarium species in the current elite maize germplasm was indicated by ECB susceptible non‐Bt hybrids with low‐MON concentrations.

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 ‐ BMDL05) 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.

Moniliformin (MON) is a highly polar, emerging Fusarium mycotoxin with a low molecular weight. It is known to exhibit potentially harmful effects on public and animal health. This study aimed to comprehensively assess the natural occurrence of MON in various foods marketed in South Korea and to perform a risk assessment. An analytical method for MON quantification using strong anion exchange clean-up combined with ultra-high-performance liquid chromatography-tandem mass spectrometry was validated across four different food matrices (white rice, sorghum, corn oil, and baby food), exhibiting excellent accuracy, precision, and sensitivity. A total of six food categories, 33 food commodities, and 253 food samples were included in this study. Maize, sorghum, Job's tears, and perilla seeds were identified as the major contributors to MON contamination. Estimated daily intake (EDI) was calculated for both mean and 95th percentile extreme dietary scenarios using upper and lower bound approaches. The highest EDI was observed in the 0-2-year and 3-6-year age groups, primarily for cereal grains. The margin of exposure (MOE) values for maize consumption ranged from 2544 to 7482. These results highlight the potential health concerns associated with MON, necessitating targeted risk management strategies.

Deoxynivalenol and other selected Fusarium toxins in Swedish wheat — Occurrence and correlation to specific Fusarium species

Sorghum (Sorghum bicolor (L.) Moench) and pearl millet (Pennisetum glaucum (L.) R.Br) are indigenous crops to the African continent. Apart from maize, rice and wheat, these crops are basic staple foods for many rural communities in Africa. The growth and production of these grains can be negatively affected by plant diseases caused by diverse fungal genera. The ability of Fusarium species to produce mycotoxins, including fumonisins (FUM) and moniliformin (MON), that have detrimental health effects for both humans and animals make it important to evaluate their toxin production in crops that are intended for human consumption. Fusarium species occur naturally in maize, sorghum and millet, among other grains. Potentially toxigenic species isolated from grain samples from Nigeria harboured high FUM and MON producing strains. This was confirmed by molecular identification and by chemoprofiling in in vitro grain cultures. Mycotoxin levels of Fusarium species grown on maize patty cultures were compared to levels produced on sorghum and millet patty cultures. FUM and MON profiles of 18 Fusarium proliferatum and two other Fusarium control strains, ie high producers of either one of these toxins, were analyzed. FUM (fumonisin B1, B2 and B3) were extracted with methanol/water and MON with acetonitrile/water. The mycotoxin extracts were cleaned up using strong anion exchange solid phase extraction prior to quantification by reversed-phase HPLC. Results indicated that under conducive conditions, all the strains tested produced FUM, some in relatively large quantities (11/18), ranging from 694-17421 mg/kg culture material. For 8/18 strains the MON levels were >500 mg/kg and up to 8892 mg/kg culture material. Although there are variations in the potential or ability of F. proliferatum isolates to produce either FUM or MON, these fungi can use several grains as a source for toxin production irrespective of their original hosts. This study gives insight into the potential and ability of Fusarium species, isolated from maize, sorghum and millet, to produce mycotoxins on several grain sources, which may have a marked influence on food safety and security, and the potential health risk they hold for many rural communities in Africa. Vismer HF, Shephard GS, Imrie G, Van der Westhuizen L, Volkwyn Y, Mngqawa P

Aquaculture feed manufacturers and producers increasingly recognize the importance of mycotoxins, which contaminate plant-based meals used in compound aquafeeds, and their potential to negatively impact production. Though data on the worldwide occurrence of legislated mycotoxins e.g., trichothecenes and zearalenone (ZEN) are well documented, relatively little information is available regarding other mycotoxins also produced by Fusarium, notably moniliformin (MON). Given that MON is known to affect the survival, growth, skeletal formation and bone mineralization in terrestrial species, its widespread occurrence on maize and maize by-products typically used in aquaculture makes it relevant to study these parameters in teleost fish. In the present work we have tested the effect of MON exposure on survival, bone development and mineralization using zebrafish (Danio rerio) as a model species and fish derived osteo-chondroprogenitor cell line for in vitro studies. Moniliformin exposure did not decrease bone mineralization in zebrafish larvae or extracellular matrix mineralization in the mineralogenic cell line VSa13. Here, the minimal in vitro cytotoxicity concentration was found to be 1000 µg L−1 MON. Incidence of deformities was also not altered by MON at the concentration tested (450 µg L−1) although larval growth was affected, as shown by a decrease in the standard length of exposed specimens at 20 days post fertilization. Survival decreased significantly in larvae exposed to MON concentrations higher than 900 μg L−1. Influence of MON on survival and growth might be relevant for aquaculture industry. As MON is a water-soluble mycotoxin, its leaching from feed is highly probable, so MON assimilation into the surrounding aqueous environment should also be considered. Tested levels in fish larvae are within the reported occurrence levels of MON in commercial feed and plant meals.

Cereal grains and their products are among the basic components of the diet all over the world. Their consumption varies depending on the dietary habits in each country. Apart from ingredients valuable from a health point of view, these products can also be a source of contaminants. The European Food Safety Authority (EFSA) included these foodstuffs in the group of main contributors to the dietary exposure to nickel by different groups of the population. The aim of the studies was to determine the nickel content in cereal grains and selected cereal products commercially available in Poland and to assess the exposure of consumers to nickel intake with these foodstuffs. A total of 56 samples of cereal grain and cereal products from trade were tested. Nickel content was determined after microwave mineralization of the samples by atomic absorption spectrometry with a graphite furnace atomization (GFAAS). The estimated exposure for the different groups of the population was assessed and compared with the Tolerable Daily Intake (TDI) value established by the European Food Safety Authority based on an updated risk assessment at 13 μg/kg body weight (b.w.) per day. The mean (MB) and 95th percentile (MB) nickel content in investigated samples of cereal grains and their products were 0.66 mg/kg and 1.93 mg/kg. The mean content of Ni in the analyzed samples of grains was 1.16 mg/kg. Obtained results in this group of products ranged from 0.10 mg/kg for rye to 4.80 mg/kg for millet. In the group of grain products, the mean (MB) concentration of nickel was 0.61 mg/kg (95th percentile (MB) 1.84 mg/kg). The highest nickel level was determined in the samples of bran, groats, and flakes compared to other grain-based products. The highest content of Ni in the group of cereal products was found in the samples of roasted buckwheat 1.81 mg/kg and oat flakes 2.53 mg/kg. Significantly lower nickel content was observed in barley groats as well as for pasta and flour. The estimated average exposure of adults and children to nickel intake with grains and grain-based products ranged from 1.1% to 13.4% of TDI and it does not pose a health risk for consumers. Based on the obtained results, it was found that the nickel content in the tested samples of cereals and their products does not pose a health risk, even though the contamination of individual samples was significant.

Corn is a vital cereal consumed globally because of its rich nutritional value. However, corn grains are prone to contamination with numerous mycotoxin-producing fungi from farm to fork. Mycotoxins are secondary metabolites that severely threaten the human health and lead to economic loss of the grain industry. Various techniques are applied at different stages after harvesting corn to remove or degrade mycotoxins. The decontamination techniques are categorized into physical (cold plasma, irradiation, and microwave), chemical (ozone, chlorine dioxide, and ammonia), and biological methods (non-toxigenic strains, lactic acid bacteria, and enzymes). These techniques have varied efficiencies for the degradation of mycotoxins from corn or corn-based products. The decontamination technologies are selected based on the maximum reduction of mycotoxins and minimum effect on corn's nutritional and bioactive properties. This review addressed the role of corn-based products in human health, mycotoxin contamination, and decontamination techniques, challenges, and possible approaches. Researchers are in the process of developing and optimizing new decontamination techniques that might reduce toxigenic fungi or mycotoxins from corn without adversely affecting corn-based products.