Occurrence of moniliformin,deoxynivalenol, and zearalenone in durum wheat (Triticum durum Desf.)

Forty-eight durum wheat samples from 5 locations in Austria were examined forFusarium infection andFusarium toxin content.F.gramlnearum andF.avenaceum were by far the prevailingFusarium species In durum wheat kernels, followed byEpoae, F.culmorum, andF.equlsetl. Ion-paired HPLC analyses of the samples showed moniliformin contents of kernels up to 0.88 mg/kg. All moniliformin contaminated samples also contained high levels of deoxynivalenol (up to 8.2 mg/kg) and lower levels of zearalenone (<0.33 mg/kg). The levels of zearalenone in naturally contaminated durum wheat samples did not correspond to the high yields of zearalenone found in cultures of the fusaria isolated from the durum wheat kernels. These conflicting results as well as some toxicologlcal aspects of the carry over ofFusarium toxins from durum wheat kernels into pasta are discussed.     

Occurrence and toxicity ofFusarium subglutinans from Peruvian maize

Twenty-five samples of maize kernels collected at harvest time from geographically different corn fields in Peru, were examined for the occurrence of toxigenicFusarium species. The most frequently recovered species wereF. subglutinans (48%),F. moniliforme (46%), andF. equiseti (5%). OtherFusarium species isolated (up to 1%) includedF. graminearum, F. acuminatum, F. solani, F. oxysporum, andF. culmorum. Assays ofFusarium culture extracts usingArtemia salina larvae, showedF. subglutinans as one of the most toxigenic species, and its toxicity was mostly correlated to the capability to produce beauvericin (BEA). All eight tested isolates ofF. subglutinans grown on autoclaved corn kernels produced BEA (from 50 to 250 mg/Kg) as well as moniliformin (M) (from 70 to 270 mg/Kg). This is the first report on BEA and M production by maize isolates ofF. subglutinans from South America.     

Production of moniliformin by Canadian isolates of Fusarium

Twenty-eight Canadian isolates of Fusarium were tested for their ability to produce moniliformin in corn. Both F. moniliforme (2/6 isolates) and F. subglutinans (11/15 isolates) produced the mycotoxin, while F. graminearum did not. Field-corn inoculated with F. moniliforme M3783 was able to support production of both moniliformin and fusarin C.     

Mycotoxin production by Fusarium species isolated from New Zealand maize fields

Forty Fusarium isolates obtained from maize fields were screened for moniliformin production on maize kernels. Twelve isolates, including seven of F. subglutinans, were found to produce moniliformin at levels ranging from 0.4 to 64 ppm. Twenty six isolates were also screened for production of deoxynivalenol, diacetoxyscirpenol, T-2 toxin and zearalenone. Of these, 22, including all 11 isolates of F. graminearum, produced zearalenone at levels ranging from 0.1 to 96.0 ppm, while 13 produced T-2 toxin at low levels, (<1.1 ppm). Deoxynivalenol and diacetoxyscirpenol were each produced by six isolates, also at low levels (<1.0 ppm). Three isolates of F. graminearum and one of F. sambucinum produced four toxins simultaneously.     

Moniliformin In wheat and triticale grain

Fusarium avanacoum infected wheat and triticale heads in Poland in each season between 1985 and 1989. The average number of heads infected byF avonacaum was 26 % for wheat and 46 % for triticale out of all examined heads withFusarium head blight symptoms.Fusarium-damaged wheat grain, naturally infected byF avenaceum, contained an average of 15.9±7.7 mg moniliformin/kg, healthy looking kernels from the same heads an average of 0.42±0.19 mg moniliformin/kg. Fusarfum-damaged kernels of triticale contained an average of 3.5 mg moniliformin/kg while healthy looking kernels from the same ears contained 0.25 mg/kg.     

Evaluation of trichothecene and nontrichothecene mycotoxins produced byFusarium in soybeans

Each of 12 cultures ofFusarium, comprising four species, isolated from moldy soybeans suspected of being involved in illness of wild geese, were grown separately in autoclaved moist rice, in autoclaved moist soybeans, and in surface sterilized-disinfected soybeans, assayed for various mycotoxins, and fed to rats. Four additional cultures that produced known toxins on rice were also grown on soybeans as controls. All isolates, except one ofF moniliforme, grown in rice resulted in weight loss of rats, and that one resulted in weight gain; 12 of the isolates caused death. One isolate ofF poae grown in soybeans caused death when consumed by rats, but none of the other 15 resulted in weight loss or overt injury. Much larger amounts of zearalenone, deoxynivalenol (DON), T-2 toxin, neosolaniol, T-2 tetraol, wortmannin, and moniliformin were produced by the cultures on rice than on soybeans, but more HT-2 toxin was produced by one isolate ofF poae grown on soybeans than when grown on rice. Soybeans appear to be a poor substrate for elaboration of most of the toxins produced by the isolates tested.     

Diversity of Fusarium species and mycotoxins contaminating pineapple

Pineapple (Ananas comosus var. comosus) is an important perennial crop in tropical and subtropical areas. It may be infected by various Fusarium species, contaminating the plant material with mycotoxins. The aim of this study was to evaluate Fusarium species variability among the genotypes isolated from pineapple fruits displaying fungal infection symptoms and to evaluate their mycotoxigenic abilities. Forty-four isolates of ten Fusarium species were obtained from pineapple fruit samples: F. ananatum, F. concentricum, F. fujikuroi, F. guttiforme, F. incarnatum, F. oxysporum, F. polyphialidicum, F. proliferatum, F. temperatum and F. verticillioides. Fumonisins B₁–B₃, beauvericin (BEA) and moniliformin (MON) contents were quantified by high-performance liquid chromatography (HPLC) in pineapple fruit tissue. Fumonisins are likely the most dangerous metabolites present in fruit samples (the maximum FB₁ content was 250 μg g^?1 in pineapple skin and 20 μg ml^?1 in juice fraction). In both fractions, BEA and MON were of minor significance. FUM1 and FUM8 genes were identified in F. fujikuroi, F. proliferatum, F. temperatum and F. verticillioides. Cyclic peptide synthase gene (esyn1 homologue) from the BEA biosynthetic pathway was identified in 40 isolates of eight species. Based on the gene-specific polymerase chain reaction (PCR) assays, none of the isolates tested were found to be able to produce trichothecenes or zearalenone.     

Production of Fumonisins by Fusarium moniliforme Cultures Isolated from Jimsonweed in Mississippi

Eight Fusarium spp. were isolated from greenhouse-grown jimsonweed (Datura stramonium L.) in Mississippi in 1990. Four isolates of Fusarium moniliforme were obtained and when grown on autoclaved rice, produced 115 to 3,200 mg/kg fumonisin B₁, (FB₁). Other fumonisin-related compounds, such as FB₂, FB₃ and FB₄ were also produced at levels of 240, 210 and 160 mg/kg, respectively. F. semitectum (1 isolates) was negative for production of fumonisin and other phyto-toxins. F. oxysporutn (1 isolate) produced only 3.5 g/kg moniliformin. This is the first report of production of fumonisins by F. moniliforme isolated from weeds such as jimsonweed.     

Toxicity and moniliformin production by four recently described species of Fusarium and two uncertain taxa

Four recently described species, Fusarium nygamai, F. dlamini, F. beomiforme and F. napiforme and two uncertain taxa, F. nygamai from millet in Africa and Fusarium species from rice with Bakanae disease, were tested for toxicity and moniliformin production. Cultures grown on autoclaved corn were fed to groups of four one-day-old ducklings for 14 days. Isolates that caused the death of 3 or 4 out of 4 ducklings were considered to be toxic and analyzed for moniliformin. All 15 isolates of F. dlamini tested were nontoxic. The other taxa contained some isolates that were toxic to ducklings and produced moniliformin in corn cultures. This is the first report of moniliformin production by F. beomiforme (200–890 g/g), and F. napiforme (16–388 g/g), and by F. nygamai not obtained from millet in Africa (15–874 g/g). The highest production of moniliformin was obtained from the 19 isolates of F. nygamai from millet in Africa (4300–18200g/g) and the 15 isolates from rice with Bakanae disease (2300–19300 g/g). The taxonomic position of these two uncertain taxa should be re-evaluated.     

Screening for Fusarin C production by European isolates of Fusarium species

A total of 137Fusarium isolates were screened forin vitro production of the mutagenic metabolite fusarin C, using a simple thin layer chromatographic method. It has been proven thatFusarium species (F. culmorum, F. graminearum, F. crookwellense, F. sporotrichioides, F. poae, F. tricinctum, andF. Avenaceum) isolated from European agricultural crops and soils are able to produce fusarin C. No fusarin C production was detected among isolates ofF. arthrosporioides, F. acuminatum, or F. equiseti. Results obtained by High-Performance Liquid Chromatography (HPLC) analyses of fungal extracts show that up to 26 chromatographic peaks having UV spectra similar to that of fusarin C are produced. It is not known if any of these metabolites are as mutagenic as fusarin C.     

Prevalence of Fusarium species of the Liseola section on Zimbabwean corn and their ability to produce the mycotoxins zearalenone,moniliformin and fumonisin B1

Maize samples were collected from nine Grain Marketing Board (G.M.B) centers in Zimbabwe during the 1991 harvest season. A further 47 samples collected directly from farmers and from the G.M.B., centers in Chinhoyi and Kwekwe during the 1992 harvest season. These samples were analyzed mycologically and the predominant flora was Fusarium although Penicillium, Nigrospora, Aspergillus and Chaetomium could be isolated from some samples. From the first nine samples studied, F. verticillioides and F. subglutinans were isolated in almost equal proportions on samples from the central and the south of the country whereas only F. verticillioides was isolated on the samples from the north. The subsequent study demonstrated that there was a greater fungal diversity in samples from North (Mashonaland West) than samples from the South (Midlands area) with species of Nigrospora, Chaetomium, Acremonium and Diplodia occurring in significant numbers. From a total of 2821 fungal isolates obtained from all the maize samples analyzed, 1485 (53%) were found to belong to the liseola section of Fusarium. The ability of these isolates to produce the mycotoxins zearalenone, moniliformin and fumonisin B₁ was tested using a simplified TLC Agar plate method. Out of the 886 isolates tested, only one produced all the three mycotoxins simultaneously whilst most produced fumonisin B₁ and/or moniliformin. Only nine isolates produced zearalenone. This revised version was published online in June 2006 with corrections to the Cover Date.     

Production of zearalenone,moniliformin and trichothecenes in intact sugar beets under laboratory conditions

Five toxigenic isolates of Fusarium species were tested for the production of zearalenone, moniliformin and trichothecenes (deoxynivalenol, 15-acetyldeoxynivalenol, T-2, HT-2 and neosolaniol) when grown on solid sugar beet slices in the laboratory for thirty days. The isolates were also grown on a solid rice medium for comparison. High zearalenone and trichothecene-producing isolates originally obtained from corn and corn-based feedstuff were compared with isolates obtained from sugar beets. One moniliformin-producing isolate from wheat was included in the study. With the exception of moniliformin, all toxins were produced on both substrates; however, the rice medium yielded the greater concentrations except for HT-2 which was produced on sugar beets in equal or greater concentrations. Zearalenone production on rice reached 729–1943 gmg/g whereas on sugar beet it reached 72–193 gmg/g. The moniliformin-producing isolate grew well on both substrates; however, moniliformin was produced only on the rice substrate. This study demonstrates for the first time that Fusarium species can produce both zearalenone and the trichothecenes on a sugar beet substrate.     

Moniliformin and the European Corn Borer (<Emphasis Type="Italic">Ostrinia nubilalis</Emphasis>)

A representative survey was made of maize ears of the 1988 and 1989 crop in Austria to establish the influence of corn borer injuries onFusarium species involved in ear fusariosis andFusarium toxin production.TheFusarium species most frequently isolated from rot-damaged ears wereFsacchari var. subglutinans (about 50 %) andF. graminearum (about 30 %). There was a striking difference between theFusarium species of the Liseola and the Discolor section concerning their occurrence on corn borer-damaged ears. More than 80 % of the ears infected withF. sacchari var. subglutinans andF. verticillioides, but less than 15 % of the ears infected withF. graminearum, F. crookwellense andF. culmorum showed corn borer injuries.Toxin analyses of the infected ears corresponded to the known toxigenicity of the respectiveFusarium species. Ears infected withF. sacchari var. subglutinans contained moniliformin (up to 20 mg/kg), those infected withF. verticillioides fumonisin B₁ and B₂ (up to 15 mg/kg). In ears infected withF. graminearum, F. culmorum andF. crookwellense zearalenone (up to 40 mg/kg) and deoxynivalenol (up to 500 mg/kg) or nivalenol (up to 10 mg/kg), respectively, could be detected. Hence measures to combat the European corn borer will mainly reduce moniliformin and fumonisin contamination, but will affect zearalenone, deoxynivalenol and nivalenol contents of the ears to a much lesser extent.     

Effects of beauvericin to mammalian tissue and its production by Austrian isolates ofFusarium proliferatum and Fusarium subglutinans

Austrian isolates ofFusarium subglutinans andFusarium proliferatum were studied for their ability to produce beauvericin, moniliformin and fumonisin B₁ and B₂ under laboratory conditions. Analytical methodology for beauvericin was specially adapted for this task. Our analyses showed that the strains produced beauvericin up to 687 mg /kg maize and moniliformin up to 70 mg/kg. The culture ofF. proliferatum in addition produced fumonisin B₁ and B₂ at levels of 106 and 61 mg/kg,respectively. The preliminary toxicity experiments performed in this study clearly indicated a toxic effect of beauvericin on the contractility of mammalian smooth muscle and thus on mammalian cells.     

Preservation of fungi in water (Castellani): 20 years

Sixty-two isolates of Fusarium were obtained from pasture grass and soil from various areas of New Zealand and identified as F. anthophilum [2], F. avenaceum [17], F. crookwellense [8], F. culmorum [4], F. graminearum [1], F. nivale [3], F. oxysporum [3], F. sambucinum [17], F. semitectum [1], F. tricinctum [1] and an unidentified Fusarium spp. [5]. These isolates were grown on autoclaved rice and tested for toxicity to rats in feeding tests. Eighty two percent of the isolates were toxic, of which twenty-four percent were severely toxic and caused hemorrhages of stomach and intestine, hematuria, and finally death. Cultures of the most toxic isolates contained 0.1 to 104 ppm of deoxynivalenol, 0.7 and 7 ppm of 15- and 3-acetyldeoxynivalenol respectively, 0.2 to 4 ppm of fusarenon- X, 11 to 1021 ppm zearalenone, 40 to 272 ppm of the hemorrhagic factor (wortmannin), 2,100 to 7,200 ppm of moniliformin, 565 ppm of the cytotoxic factor (HM-8) and enniatin in substantial concentrations. F. sambucinum is reported as a moniliformin producer for the first time.     

Bikaverin production by Fusarium species

Production of bikaverin has been examined in 130Fusarium isolates belonging to 21 species. The highest yield of bikaverin was produced on autoclaved rice — up to 2.5g/kg of dry culture. Bikaverin was produced by the following species:F verticillioides, F sacchari varsubglutinans, F proliferatum, F anthophilum, F oxysporum, F dlamini, F nygamai, F napiforme, andF solani. SpeciesF coeruleum, F poae, F sporotrichioides, F tricinctum, F chlamydosporum, F culmorum, F graminearum, F cerealis (F crookwellense), F avenaceum, F acuminatum, andF equiseti did not produce bikaverin. The production of bikaverin determines the colour of the mentionedFusarium species cultures on agar media and on rice. The pigment has indicator properties and changes colour from red in acidic solution to violet-blue in alkaline. The role it plays in fungus metabolism is not elucidated.     

Production of zearalenone,nivalenol, moniliformin,and wortmannin from toxigenic cultures ofFusarium obtained from pasture soil samples collected in New Zealand

One culture ofF avenaceum, 4 cultures ofF oxysporum, and 11 cultures of Fsambucinum were isolated from soil samples of pasture in New Zealand in 1987. All cultures, when grown on rice media and fed to rats caused a weight loss in rats as well as toxic signs including hemorrhaging and congestion, uterine enlargement, and hematuria. 6 out of 16 cultures caused death in rat feeding tests.F oxysporum #1 killed rats (feeding test) within 5-12hrs. 10 cultures produced zearalenone (19 to 8,849 ppm), 8 cultures produced nivalenol (32 to 117 ppm), 1 culture,F sambucinum #8, produced wortmannin (40 ppm), and 5 cultures produced moniliformin (19 to 9,000ppm). We report for the first time the co-occurrence of zearalenone, nivalenol, and moniliformin produced byF sambucinum #3 in culture.F avenaceum #1 andF oxysporum cultures (nos 1, 2, and 3) produced moniliformin alone.F oxysporum #4 produced zearalenone alone as well.F sambucinum #5 caused erythema in the small intestine of rats and 100% mortality and did not produce any known toxin(s). Nivalenol when administered to the stomach of rats orally at levels 10, 20, and 40mg/kg body weight caused inflammation in the intestines, coma, and death. The mycotoxins T-2 toxin, HT-2 toxin, T-2 tetraol, diacetoxyscirpenol (DAS), monoacetoxyscirpenol (MAS), deoxynivalenol (DON), 3-acetyl-and 15-acetyldeoxynivalenol, depoxynivalenol, fusarenon-X, alpha-and beta-zearalenone, and fusarochromanone (TDP-1) were not detected in the extracts of these cultures.     

Incidence ofFusaria and occurrence of selected Fusarium mycotoxins on Lolium spp. in Germany

Test plantings with varieties ofLolium multiflorum andL perenne were harvested 4 to 7 times a year in 1991 and 1992. Samples were checked for the presence ofFusaria, the mycotoxins zearalenone, T-2 toxin, and diacetoxyscirpenol (DAS). Spectrum of species and the incidence ofFusaria and fusariotoxins are discussed in relation to the influencing factors site, variety ofLolium, harvesting time and year. Depending on these factors, 41 % to 100 % of the samples wereFusarium positive. Differences in infestation with Fusarium among varieties ofLolium perenne were dependent on location and did not correlate with yield. The six species ofFusarium pathogenic toLolium spp. (F. graminearum, F. culmorum, F. avenaceum, F. oxysporum, F. solani, and F. acuminatum) totaled 35.7 % of all the isolated strains. 14 species could be isolated fromLolium samples (descending frequency):F. culmorum, F. sambucinum, F. equiseti, F. acuminatum, F. semitectum, F. oxysporum, F. subglutinans, F. avenaceum, F. sporotrichioides, F. proliferatum, F. tricinctum, F. anthophilum, F. dimerum and F. graminearum. For the detection ofFusaria a promising new immunological method is presented. It is based on the genus specific production of exopolysaccharides byFusarium species. Mycotoxin contents in grass ranged from 0.01 to 4.75 ppm for zearalenone with 67 % positive samples and 0.3 % samples above 1 ppm, 0.04 to 2.78 ppm for T-2 toxin with 25 % positive samples and 2.8 % samples above 1 ppm, and 0.003 to 0.06 for DAS with 21.6 % positive samples. In silages, no T-2 toxin was detectable. IsolatedFusarium strains were checked for the ability to produce the mycotoxins zearalenone, T-2 toxin and DAS in culture. Most of the strains were positive for at least one of the toxins.     

Moniliformin production and toxicity of different Fusarium species from Southern Africa.

Four new moniliformin-producing species of Fusarium were found, viz., F. acuminatum, F. concolor, F. equiseti, and F. semitectum. Isolates of F. acuminatum and F. concolor produced large amounts of moniliformin (3.4 and 9.5 g/kg, respectively), whereas isolates of the other three species yielded less than 30 mg/kg. The production of moniliformin by isolates of F. oxysporum and F. avenaceum from southern Africa is described. All 14 toxic isolates of F. oxysporum produced moniliformin. Most isolates of F. fusarioides and all six isolates of Fusarium moniliforme va. subglutinans tested produced moniliformin, as did 28 of 36 toxic isolates of F. moniliforme. A number of F. moniliforme isolates produced greater than 10 g/kg, and one isolate yielded 33.7 g/kg in corn after incubation for 5 weeks at 25 degrees C. Moniliformin production in the field in corn ears was shown by inoculating plants with known moniliformin-producing isolates of three Fusarium species. Yields of up to 645 mg/kg were recorded. Isolates of F. acuminatum, F. equiseti, F. fusarioides, and F. moniliforme were found that were highly toxic to ducklings but which did not produce moniliformin.     

A Hemorrhagic Factor (Apicidin) Produced by Toxic Fusarium Isolates from Soybean Seeds

Fifty-two isolates of Fusarium species were obtained from soybean seeds from various parts of Korea and identified as Fusarium oxysporum, F. moniliforme, F. semitectum, F. solani, F. graminearum, or F. lateritium. These isolates were grown on autoclaved wheat grains and examined for toxicity in a rat-feeding test. Nine cultures were toxic to rats. One of these, a culture of Fusarium sp. strain KCTC 16677, produced apicidin, an antiprotozoal agent that caused toxic effects in rats (including body weight loss; hemorrhage in the stomach, intestines, and bladder; and finally death) when rats were fed diets supplemented with 0.05 and 0.1% apicidin. The toxin was toxic to brine shrimp (the 50% lethal concentration was 40 μg/ml) and was weakly cytotoxic to human and mouse tumor cell lines.