Concentrations Of Moniliformin Research Articles

Regional and field-specific differences in Fusarium species and mycotoxins associated with blighted North Carolina wheat

Worldwide, while Fusarium graminearum is the main causal species of Fusarium head blight (FHB) in small-grain cereals, a diversity of FHB-causing species belonging to different species complexes has been found in most countries. In the U.S., FHB surveys have focused on the Fusarium graminearum species complex (FGSC) and the frequencies of 3-ADON, 15-ADON, and nivalenol (NIV) chemotypes. A large-scale survey was undertaken across the state of North Carolina in 2014 to explore the frequency and distribution of F. graminearum capable of producing NIV, which is not monitored at grain intake points. Symptomatic wheat spikes were sampled from 59 wheat fields in 24 counties located in three agronomic zones typical of several states east of the Appalachian Mountains: Piedmont, Coastal Plain, and Tidewater. Altogether, 2197 isolates were identified to species using DNA sequence-based methods. Surprisingly, although F. graminearum was the majority species detected, species in the Fusarium tricinctum species complex (FTSC) that produce “emerging mycotoxins” were frequent, and even dominant in some fields. The FTSC percentage was 50–100% in four fields, 30–49% in five fields, 20–29% in five fields, and < 20% in the remaining 45 fields. FTSC species were at significantly higher frequency in the Coastal Plain than in the Piedmont or Tidewater (P < .05). Moniliformin concentrations in samples ranged from 0.0 to 38.7 μg g−1. NIV producing isolates were rare statewide (2.2%), and never >12% in a single field, indicating that routine testing for NIV is probably unnecessary. The patchy distribution of FTSC species in wheat crops demonstrated the need to investigate the potential importance of their mycotoxins and the factors that allow them to sometimes outcompete trichothecene producers. An increased sampling intensity of wheat fields led to the unexpected discovery of a minority FHB-causing population.

Preliminary Evaluation of Moniliformin as a Potential Threat for Teleosts

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.

Fusarium Damage in Cereal Grains from Western Canada. 1. Phylogenetic Analysis of Moniliformin-Producing Fusarium Species and Their Natural Occurrence in Mycotoxin-Contaminated Wheat, Oats, and Rye

Harvest samples of common wheat (Triticum aestivum), oats (Avena sativa), and rye (Secale cereale) from producers in western Canada were analyzed for fungal infection by toxigenic Fusarium species and contamination by trichothecenes and moniliformin (MON). Fusarium graminearum and F. avenaceum were the two most frequently isolated species from samples of rye and wheat collected in 2010. F. poae and F. sporotrichioides were more commonly detected in randomly selected oat seeds. Other toxigenic Fusarium species including F. acuminatum, F. culmorum, and F. pseudograminearum as well as Phaeosphaeria nodorum (a.k.a. Septoria nodorum) were recovered primarily from fusarium-damaged kernels of wheat. Pure cultures of F. avenaceum, F. acuminatum, and other related species known to produce moniliformin were isolated from incubated seeds based on micro- and macromorphological criteria. The phylogenetic analysis inferred from partial DNA sequences of the acl1 and tef-1α genes revealed two major clades representing F. avenaceum and F. acuminatum, respectively. These clades comprised all Canadian isolates of the two species and a number of reference cultures studied earlier for their propensity to form moniliformin in vitro and in planta. However, some reference cultures previously reported to produce significant amounts of moniliformin formed minor phylogenetic lineages that represent rather distinct but closely related species. Concomitantly, cereal samples were analyzed for the presence of deoxynivalenol and moniliformin. These two Fusarium toxins were observed most frequently in common wheat, at concentrations up to 1.1 and 4.0 mg/kg, respectively. There was no apparent relationship between moniliformin concentrations and detection of F. avenaceum and F. acuminatum in rye and oat samples. Geographical analysis of the distribution of moniliformin and F. avenaceum and F. acuminatum across the Canadian Prairies also did not indicate a strong relationship.

Fusarium Damage in Small Cereal Grains from Western Canada. 2. Occurrence of Fusarium Toxins and Their Source Organisms in Durum Wheat Harvested in 2010

Samples of Canadian western amber durum harvested in 2010 were obtained as part of the Canadian Grain Commission Harvest Sample Program, inspected, and graded according to Canadian guidelines. A subset of Fusarium -damaged samples were analyzed for Fusarium species as well as mycotoxins associated with these species, including deoxynivalenol and other trichothecenes, moniliformin, enniatins, and beauvericin. Overall, Fusarium avenaceum and F. graminearum were the top two most frequently recovered species. Phaeosphaeria nodorum (a.k.a. Septoria nodorum ), F. culmorum , F. poae , F. acuminatum , and F. sporotrichioides were observed in samples as well. All samples analyzed for mycotoxins contained quantifiable concentrations of enniatins, whereas beauvericin, deoxynivalenol, and moniliformin were measured in approximately 75% of the samples. Concentrations in Fusarium -damaged samples ranged from 0.011 to 34.2 mg/kg of enniatins plus beauvericin, up to 4.7 mg/kg of deoxynivalenol, and up to 6.36 mg/kg of moniliformin. Comparisons of enniatins, beauvericin, and moniliformin concentrations to the occurrence of various Fusarium species suggest the existence of an infection threshold above which these emerging mycotoxins are present at higher concentrations. The current grading factor of Fusarium -damaged kernels manages concentrations of these emerging mycotoxins in grain; lower provisional grades were assigned to samples that contained the highest concentrations of enniatins, beauvericin, and moniliformin.

Mango Malformation: I. Toxin Production Associated with Fusarium Pathogens

Eight Fusarium species i.e. F. subglutinans, F. solani, F. oxyspoum, F. sterilihyphosum, F. proliferatum, F. monili-forme, F. avena and F. chlamydspore isolated from mango malformed disease were tested for their ability to cause mango malformation disease and their production of moniliformin and total fumonisins (FB1 + FB2) using HPLC. A evaluated for moniliformin production, seven isolates were toxin producers, the production levels ranging from 0.51 to 8.90 µg/ml. The higher levels were produced by Fusarium subglutinans (8.51 µg/ml). Moderate concentrations of moniliformin was produced by F.moniliforme (6.90 µg/ml), F. oxysporum (6.30 µg/ml), F. proliferatum (4.10 μg/ml) and F. sterilihyphosum (1.10 μg/ml). Separation and identification of Fumonisin that was isolated from the pathogen- causing disease are made by (HPLC). A evaluated for total fumonisin production (FB1 + FB2), seven isolates were toxin producers, the production levels ranging from 0.10 to 8.30 µg/ml. The higher levels were produced by F. monili-forme (8.30 µg/ml. Moderate concentrations of fumonisin was produced by F .proliferatum (0.64 µg/ml) and F. subglutinans (0.50 µg/ml). Strong positive correlations between moniliformin and total fumonisins (FB1 + FB2) activities and malformation disease incidence by F. subglutinans, F. solani, F. oxyspoum, F. sterilihyphosum, F. proliferatum was observed.

Synthesis and evaluation of molecularly imprinted polymers as sorbents of moniliformin

Moniliformin is a low molecular weight mycotoxin that has worldwide potential to contaminate cereal grains. Although several traditional methods have been developed to detect moniliformin, the lack of anti-moniliformin antibodies has created a need for materials that recognize moniliformin at the molecular level through a binding mechanism. To address this issue, the authors synthesized molecularly imprinted polymers that bind moniliformin. Imprinted and non-imprinted polymers were evaluated by equilibrium binding assays and moniliformin concentrations were measured by LC analysis using ultraviolet light detection. Successful polymers were imprinted with toxin analogues as the templates; non-imprinted polymers exhibited minimal binding in acetonitrile under the assay conditions. Selected imprinted polymers also bound moniliformin in ethanol, methanol and dimethyl formamide. Significant differences in moniliformin binding by the polymers were dependent on polymer composition, and these differences were highly dependent on the template used to imprint the polymer. Polymers were further evaluated as sorbents for molecularly imprinted solid-phase extraction (MISPE), and an imprinted polymer was used for preconcentration and clean-up of a moniliformin spiked corn extract.

Moniliformin in Norwegian grain

Norwegian grain samples (73 oats, 75 barley, 83 wheat) from the 2000–02 growing seasons were examined for contamination with moniliformin, and the association between the fungal metabolite and the number of kernels infected with common Fusaria was investigated. Before quantification of moniliformin using ion pairing reversed-phase high-performance liquid chromatography with diode array ultraviolet light detection, all samples were extracted using acetonitrile/water (84/16) and disposable strong anion exchange columns used for clean up. The limit of detection was 40 μg kg−1. Moniliformin was found in 25, 32 and 76% of the barley, oats and wheat samples, respectively. The maximum concentrations of moniliformin in barley, oats and wheat were 380, 210 and 950 μg kg−1, respectively. At the same time, the prevalence and infection level of the moniliformin-producing F. avenaceum/arthrosporioides was as high as 100 and >53% on average, respectively. Moniliformin concentrations were significantly correlated to the variables grain species, growing season and infection with F. avenaceum/arthrosporioides and F. culmorum. The survey indicates that the prevalence of moniliformin in Norwegian grain is high, especially in wheat. On the other hand, field conditions in Norway do not seem to favour contamination of grain with high levels of moniliformin.

Concentration of moniliformin produced by Fusarium species in grains of transgenic Bt maize hybrids compared to their isogenic counterparts and commercial varieties under European corn borer pressure

AbstractThe 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.

Toxicity of Moniliformin and Fumonisin B1 Fed Singly and in Combination in Diets for Young Channel Catfish Ictalurus punctatus

Abstract.— Growth, histological lesions, and biochemical changes were investigated in channel catfish Ictalurus punctatus fed various concentrations of moniliformin with or without fumonisin B1. Channel catfish (average initial weight, 1.5 g) were fed diets formulated to contain 0, 20, 40, 60, and 120 mg moniliformin/kg; 0, 20, and 40 mg fumonisin B1/kg, or two combinations of moniliformin and fumonisin B1 for 10 wk. Fish fed diets with the lowest concentration of moniliformin or fumonisin B1 (20 mg/kg diet) had significantly (P &lt; 0.05) less weight gain than the control fish. Increasing the level of moniliformin in the diets resulted in a linear decrease in weight gain. Overall mortality of fish was 4% and not related to treatment effects. Hematocrit was significantly (P &lt; 0.05) lowered by 60‐mg moniliformin/kg diet or 40‐mg fumonisin B1/kg diet. Dose‐dependent increases in serum pyruvate concentration and ratio of free sphinganine to free sphingonine were obtained with increasing concentration of dietary moniliformin and fumonisin B1, respectively. Mean serum pyruvate level was significantly (P &lt; 0.05) higher in fish fed the diet containing 60‐mg moniliformin/kg diet. Addition of fumonisin B1 (40 mg/kg) to the diet containing 40‐mg moniliformin/kg significantly increased the serum pyruvate level above that of the control. Also, the lowest concentration of fumonisin B1 (20 mg/kg diet) significantly (P &lt; 0.05) increased the ratio of sphingolipids. Combinations of moniliformin and fumonisin B1 at levels of 20:40 and 40:40 mg/kg diet did not significantly change the effect of fumonisin B1 on the ratio of sphingolipids. The only tissue lesions observed in liver and heart were smaller nuclei of cells in livers of fish fed diets containing the two highest levels of moniliformin and the combinations of the two toxins.

Determination of the mycotoxin moniliformin in cultures of Fusarium subglutinans and in naturally contaminated maize by high-performance liquid chromatography–atmospheric pressure chemical ionization mass spectrometry

A LC–MS method employing triethylamine as ion-pairing reagent for the determination of moniliformin in culture material and naturally contaminated maize samples is described. Mass spectrometric detection of moniliformin was accomplished following atmospheric pressure chemical ionization to yield the deprotonated molecular ion [M−H] − at m/z 97. The moniliformin response was found to be linear over the injected range 10 ng to 700 ng and a detection limit of 10 ng was attainable at a signal-to-noise ( S/N) ratio of 4. Five South African strains of Fusarium subglutinans were grown on maize kernels and moniliformin extracted with an acetonitrile–water (95:5) mixture. Following sample clean up with reversed-phase (C 18) solid-phase extraction cartridges, the extracts were subjected to LC–MS analysis. Triethylamine was used as an ion-pair reagent and found to improve the retention characteristics of moniliformin without any detrimental effects to the instrument. Moniliformin concentrations ranged between 130 mg/kg and 1460 mg/kg culture. Application of this method to naturally contaminated maize samples from Transkei showed that it was capable of measuring moniliformin levels down to 10 μg/kg in selected moldy maize cobs. This is the first report on the application of LC–MS to the analysis of moniliformin in cultures of F. subglutinans and in naturally contaminated maize.

Subacute and reproductive effects in mink from exposure to Fusarium fujikuroi culture material (M-1214) containing known concentrations of moniliformin.

This study was conducted to ascertain the subacute and reproductive effects in mink (Mustela vison) resulting from exposure to moniliformin, a toxic mycotoxin produced by Fusarium fungi. In a preliminary trial, adult mink were presented diets that contained targeted concentrations of 10, 20, 40, 80, 160, or 240 ppm moniliformin provided by F. fujikuroi culture material (M-1214). The mink fed diets that contained more than 40 ppm moniliformin refused to eat significant quantities of feed. Feeding adult mink diets that contained 8.1 or 17.0 ppm (wet weight) moniliformin, provided by F. fujikuroi culture material, in a 30-day subacute trial produced no significant adverse effects on feed consumption, body weights, hematologic parameters, or serum chemical values, and notable histologic changes in tissues that were examined. In the reproduction trial, female mink were exposed to the same dietary concentrations of moniliformin provided by F. fujikuroi culture material as in the subacute test from 2 weeks prior to the breeding season until their offspring (kits) were 8 weeks old. Consumption of the high-dose (17 ppm) diet resulted in significant neonatal mortality and reduced kit body weights at birth and at 8 weeks of age. Necropsy of 8-week-old kits from the control and high-dose groups revealed no gross or histologic lesions or alterations in liver, lung, or heart tissues that could account for the mortality observed in the kits exposed to the culture material. These results indicate that long-term (105-135 days) dietary exposure to F. fujikuroi culture material containing 17 ppm moniliformin is not lethal to adult female mink, but can have adverse effects on neonatal mink.

A Sudden Death Syndrome Induced in Poults and Chicks Fed Diets Containing Fusarium fujikuroi with Known Concentrations of Moniliformin

A sudden death syndrome was induced in chicks and poults fed diets containing Fusarium fujikuroi, formulated to contain 0-330 mg/kg moniliformin (M) with or without the maximum recommended therapeutic concentration of monensin. Lesions of monensin toxicosis were not observed. Clinical signs were referable to cardiac dysfunction (sudden death, dyspnea, cyanosis, depression). Poults and chicks dying early in the study had no gross lesions or had lesions of right ventricular dilation. Treated poults and chicks dying late in the study or euthanatized at termination of the study had lesions of bilateral myocardial hypertrophy, usually concentric. Absolute heart weights and relative heart weights, expressed as a percentage of body weight, were significantly greater in treated birds than controls (P < 0.05), whereas body weights were significantly less (P < 0.05). Microscopically, lesions progressed from acute myocardial degeneration to necrosis, fibrosis, and hypertrophy. Ultrastructural findings were consistent with the gross and microscopic lesions. Serum pyruvate concentrations were a useful indicator of M-induced cardiotoxicosis. Concentrations of serum pyruvate increased with increased concentration of dietary M, but were not affected by addition of monensin to the diet. In chicks ingesting 40-300 mg/kg M, serum pyruvate concentrations were significantly greater (P > 0.05) than those in controls (controls, 0.28 +/- 0.08 mmol/liter; exposed 0.38 +/- 0.11-0.55 +/- 0.13 mmol/liter). Poults ingesting 80-330 mg/kg M had significantly greater serum pyruvate concentrations than controls (controls 0.33 +/- 0.09 mmol/liter; exposed 0.43 +/- 0.13-1.00 +/- 0.006 mmol/liter). The Vetronics System was used to evaluate electrocardiographic alterations in a limited number of chicks and poults surviving to the end of the feeding trial. Electrocardiographic alterations in poults and chicks fed diets containing > or = 40 mg/kg and > or = 160 mg/kg M, respectively, were consistent with ventricular hypertrophy, myocardial injury, and hypoxia. Electrocardiographic alterations were more striking in poults than in chicks. Altered myocardial metabolism due to M toxicosis, in conjunction with the unusual susceptibility of domestic poultry to altered cardiac metabolism, is believed to be the cause of the organ-specific lesions in these birds. These findings suggest that cardiac injury with subsequent alterations in cardiac electrical conductance may be a cause of the sudden deaths observed in poultry chronically intoxicated with dietary M.

Effects of selected secondary metabolites of Fusarium moniliforme on unscheduled synthesis of DNA by rat primary hepatocytes

The Fusarium moniliforme mycotoxins—fusarin C 1 fumonisin B 1, moniliformin and bikaverin—were evaluated for genotoxicity by their ability to induce unscheduled DNA synthesis (UDS) in primary rat hepatocytes. Isolated hepatocytes were exposed to several concentrations of moniliformin (5.0–500 μ m), bikaverin (1.0–500 μ m), fumonisin B 1 (0.5–250 μ m), or fusarin C (1.0–100 μ m). Aflatoxin B 1, a known inducer of UDS, was included as a positive control. UDS was determined by autoradiography of cells after their exposure to [ 3H]thymidine. The highest doses of fusarin C and bikaverin caused cell death, but no cytotoxicity was observed in cells exposed to moniliformin or fumonisin B 1. Fumonisin B 1, moniliformin and bikaverin were not genotoxic in the UDS assay. The results of the UDS assay with fusarin C were inconclusive since a marginal effect on UDS was obtained.

A molecular mechanism for the toxic action of moniliformin, a mycotoxin produced by fusarium moniliforme

Molecular mechanisms of the toxic action of moniliformin, an extremely toxic fungal metabolite produced by Fusarium moniliforme, were investigated. Exceedingly low concentrations of moniliformin (<5 μM) selectively inhibited mitochondrial pyruvate and α-ketoglutarate oxidations by 50 per cent. It is suggested that these inhibitory effects could constitute the major molecular mechanism of toxic action and could largely, if not exclusively, account for the clinical symptoms of moniliformin poisoning.