Fumonisin B<Subscript>1</Subscript>, a sphingoid toxin,is a potent inhibitor of the plasma membrane H<Superscript>+</Superscript>-ATPase

Fumonisin B₁ (FB₁) is an amphipathic toxin produced by the pathogenic fungus Fusarium verticillioides which causes stem, root and ear rot in maize (Zea mays L.). In this work, we studied the action of FB₁ on the plasma membrane H⁺-ATPase (EC 3.6.1.34) from germinating maize embryos, and on the fluidity and lipid peroxidation of these membranes. In maize embryos the toxin at 40 M inhibited root elongation by 50% and at 30 M decreased medium acidification by about 80%. Irrespective of the presence and absence of FB₁, the H⁺-ATPase in plasma membrane vesicles exhibited non-hyperbolic saturation kinetics by ATPH-Mg, with Hill number of 0.67. Initial velocity studies revealed that FB₁ is a total uncompetitive inhibitor of this enzyme with an inhibition constant value of 17.5±1 M. Thus FB₁ decreased V_max and increased the apparent affinity of the enzyme for ATP-Mg to the same extent. Although FB₁ increased the fluidity at the hydrophobic region of the membrane, no correlation was found with its effect on enzyme activity, since both effects showed different FB₁-concentration dependence. Peroxidation of membrane lipids was not affected by the toxin. Our results suggest that, under in vivo conditions, the plasma membrane H⁺-ATPase is a potentially important target of the toxin, as it is inhibited not only by FB₁ but also by its structural analogs, the sphingoid intermediates, which accumulate upon the inhibition of sphinganine N-acyltransferase by this toxin.     

Aromatic plants essential oils activity on <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> Fumonisin B<Subscript>1</Subscript> production in corn grain

The minimum inhibitory concentration (MIC) of Origanum vulgare, Aloysia triphylla, Aloysia polystachya and Mentha piperita essential oils (EOs) against Fusarium verticillioides M 7075 (F. moniliforme, Sheldon) were assessed, using the semisolid agar antifungal susceptibility (SAAS) technique. O. vulgare, A. triphylla, A. polystachya and M. piperita EOs were evaluated at final concentrations of 10, 20, 40, 50, 100, 200, 250, 500, 1000 and 1500 εl per litre (εl/l) of culture medium. A. triphylla and O. vulgare EOs showed the highest inhibitory effects on F. verticillioides mycelial development. This inhibition was observed at 250 and 500 εl/l for EOs coming from Aloysia triphylla and O. vulgare, respectively. Thus, the effects of EOs on FB₁ production were evaluated using corn grain (Zea mays) as substrate. The EOs were inserted on the 5th, 10th, 15th and 20th day of maize postinoculation with a conidia suspension of F. verticillioides. O. vulgare and A. triphylla were applied to give final concentrations of 30 ppm and 45 ppm, respectively. Different effects were observed in the toxicogenicity at the 20th day treatment. The O. vulgare EO decreased the production level of FB₁ (P < 0.01) while A. triphyla EO increased it (P < 0.001) with respect to those obtained in the inoculated maize, not EOs treated. Results obtained in the present work indicate that fumonisin production could be inhibited or stimulated by some constituents of EOs coming from aromatic plants. Further studies should be performed to identify the components of EOs with modulatory activity on the growth and fumonisins production of Fusarium verticillioides.     

Effect of<Emphasis Type="Italic">in vitro</Emphasis> digestion on fumonisin B<Subscript>1</Subscript> in corn flakes

Low levels of fumonisins have been found frequently in corn based breakfast cereals and can occur bound to protein and other matrix components.In vitro digestion of two samples of corn flakes was carried out under "fed conditions." Fumonisins were measured as o-phthaldialdehyde/mercaptoethanol derivatives by LC-fluorescence. One sample of corn flakes (FN12) had high concentrations of fumonisin B₁ (FB) (average 125 ng/g) and total bound FB₁, (TB FB₁) (average 92 ng/g) and the other (FN11) had a low level of free FB₁ (average 29 ng/g) and no detectable TB FB₁. After incubation of the samples with gastrointestinal tract solutions simulating saliva plus stomach and duodenal juices, chyme was analysed for FB₁, hydrolyzed FB₁ (HFB₁) and partially hydrolyzed fumonisin B₁ (PHFB₁). The bioaccessibility (percentage of FB₁ released from corn flakes into chyme) was 38-78% for incurred FB₁ in FN12, 8-54% for incurred plus spiked FB₁ in FN12, and 19-66% for incurred plus spiked FB₁ in FN11. HFB₁ and PHFB₁ were not detected. If free FB₁ was first extracted from sample FN12, no FB1 was detected in the chyme, indicating no contribution from TB FB₁. Concentrations were corrected for method recovery of FB₁ or, for bound FB₁, partial method recovery of HFB₁ Presented at the XIIth IUPAC International Symposium on Mycotoxins and Phycotoxins, Istanbul, Turkey, 21–25 May, 2007     

Biomolecule-functionalized magnetic nanoparticles for flow-through quartz crystal microbalance immunoassay of aflatoxin B<Subscript>1</Subscript>

A flow-through quartz crystal microbalance (QCM) immunoassay method has been developed based on aflatoxin B₁ antibody (anti-AFB₁)-functionalized magnetic core-shell Fe₃O₄/SiO₂ composite nanoparticles (bionanoparticles) in this study. To construct such an assay protocol, anti-AFB₁, as a model protein, was initially covalently immobilized onto the Fe₃O₄/SiO₂ surface, and then the functionalized nanoparticles were attached to the surface of the QCM probe with an external magnet. The binding of target molecules onto the immobilized antibodies decreased the sensor’s resonant frequency, and the frequency shift was proportional to the AFB₁ concentration in the range of 0.3–7.0 ng/ml. The regeneration of the developed immunosensor was carried out via attaching or detaching the external magnet from the detection cell. In addition, the selectivity, reproducibility, and stability of the proposed immunoassay system were acceptable. Compared with the conventional ELISAs, the proposed immunoassay system was simple and rapid without multiple labeling and separation steps. Importantly, the proposed immunoassay method could be further developed for the immobilization of other antigens or biocompounds.     

Fungi,Aflatoxins, Fumonisin B<Subscript>l</Subscript> and Zearalenone Contaminating Sorghum-based Traditional Malt,Wort and Beer in Botswana

Brewing and consumption of traditional beer have social–economic significance in most African countries including Botswana. Traditional sorghum malt, wort, and beer samples were collected from three villages around Gaborone, Botswana. Forty-six malt samples were analyzed for fungi on three different media and developing colonies were subcultured for identification. Rhizopus, Fusarium, Mucor, and Aspergillus were the most common genera isolated. Out of the 46 malt samples, 72% contained Rhizopus stolonifer, 63% Fusarium verticillioides (syn. Fusarium moniliforme), and 37% Aspergillus flavus. Although Aspergillus flavus was isolated from malt samples, aflatoxins (B₁, B₂, G₁, and G₂) were not detected in any of the samples analyzed. When the malt, wort, and beer samples were analyzed for fumonisin B_l and zearalenone, fumonisin B₁ was detected in 3 malt samples, with concentrations ranging from 47 to 1316 μg/kg, while zearalenone was detected in 56%, 48% and 48% of the malt, wort and beer samples, respectively. Zearalenone concentration in samples ranged from 102 to 2213 μg/kg in malt, 26 to 285 μg/l in wort and 20 to 201 μg/l, in beer. Zearalenone carry-over from wort to beer ranged from 23 to 403%. Therefore, although aflatoxins and fumonisin B₁ do not appear to be major contaminants, zearalenone is common and could pose a potential problem in traditional beer in Botswana.     

GABA<Subscript>B</Subscript> Receptors in Neuroendocrine Regulation

Gamma-amino butyric acid (GABA), in addition to being a metabolic intermediate and the main inhibitory neurotransmitter in the synaptic cleft, is postulated as a neurohormone, a paracrine signaling molecule, and a trophic factor. It acts through pre- and post-synaptic receptors, named GABA_A and GABA_C (ionotropic receptors) and GABA_B (metabotropic receptor). Here we reviewed the participation of GABA_B receptors in the regulation of the hypothalamic-pituitary-gonadal axis, using physiological, biochemical, and pharmacological approaches in rats, as well as in GABA_B1 knock-out mice, that lack functional GABA_B receptors. Our general conclusion indicates that GABA_B receptors participate in the regulation of pituitary hormone secretion acting both in the central nervous system and directly on the gland. PRL and gonadotropin axes are affected by GABA_B receptor activation, as demonstrated in the rat and also in the GABA_B1 knock-out mouse. In addition, hypothalamic and pituitary GABA_B receptor expression is modulated by steroid hormones. GABA participation in the brain control of pituitary secretion through GABA_B receptors depends on physiological conditions, being age and sex critical factors. These results indicate that patients receiving GABA_B agonists/antagonists should be monitored for possible endocrine side effects.     

The short-rooted vitamin B<Subscript>6</Subscript>-deficient mutant <Emphasis Type="Italic">pdx1</Emphasis> has impaired local auxin biosynthesis

The phytohormone auxin regulates many aspects of plant growth and development. Auxin often acts distantly from the site of its biosynthesis and this long-distance-transported auxin is well known to play a critical role in eliciting physiological responses including regulating root development. Auxin can be produced in roots, yet the function of locally synthesized auxin in root growth is unclear. The major auxin in plants, indole 3-acetic acid (IAA), is mainly synthesized through tryptophan (Trp)-dependent pathways that require pyridoxal phosphate (an active form of vitamin B₆)-dependent enzymes. We previously reported that the Arabidopsis vitamin B₆ biosynthesis mutant pdx1 has stunted root growth although the underlying cause is unknown. Here we showed that the pdx1 root is deficient in auxin biosynthesis. By reciprocal grafting of pdx1 and the wild type, we demonstrated that the stunted root growth in pdx1 is caused by a locally generated signal(s) in roots. To test whether auxin might be one such signal, the auxin responsive DR5::GUS reporter was introduced into the mutant. The DR5::GUS activity in pdx1 root tips was greatly reduced compared with that in the wild type although the auxin response was unaltered. pdx1 also suppresses the root hair growth defects in the auxin overproduction mutant yucca. These data indicate that pdx1 is impaired in Trp-dependent auxin biosynthesis, which may contribute to the short-root phenotype of pdx1. We suggest that locally synthesized auxin may play a critical role in postembryonic root growth.     

Mitochondrial and cell-surface F<Subscript>0</Subscript>F<Subscript>1</Subscript>ATPsynthase in innate and acquired cardioprotection

Mitochondria are central to heart function and dysfunction, and the pathways activated by different cardioprotective interventions mostly converge on mitochondria. In a context of perspectives in innate and acquired cardioprotection, we review some recent advances in F₀F₁ATPsynthase structure/function and regulation in cardiac cells. We focus on three topics regarding the mitochondrial F₀F₁ATPsynthase and the plasma membrane enzyme, i.e.: i) the crucial role of cardiac mitochondrial F₀F₁ATPsynthase regulation by the inhibitory protein IF₁ in heart preconditioning strategies; ii) the structure and function of mitochondrial F₀F₁ATPsynthase oligomers in mammalian myocardium as possible endogenous factors of mitochondria resistance to ischemic insult; iii) the external location and characterization of plasma membrane F₀F₁ ATP synthase in search for possible actors of its regulation, such as IF₁ and calmodulin, at cell surface.     

Co-occurrence of aflatoxin B<Subscript>1</Subscript> and cyclopiazonic acid in sour lime (<Emphasis Type="Italic">Citrus aurantifolia Swingle</Emphasis>) during post-harvest pathogenesis by <Emphasis Type="Italic">Aspergillus flavus</Emphasis>

During hot and humid seasons, extensive rot of sour lime was observed to be caused by Aspergillus flavus. In view of this, investigations were undertaken to obtain data on the production of various toxins by A. flavus during post harvest pathogenesis of sour lime. Sixty percent of the pathogenic A. flavus isolates were detected to be aflatoxin B₁ producers in sour lime tissue. It was also noted that thirty three percent of aflatoxigenic A. flavus isolates had the potential to coproduce cyclopiazonic acid (CPA). Such aflatoxigenic isolates produced quantitatively more CPA (ranging from 250.0 to 2501.3 g/kg) than aflatoxin B₁ (ranging from 141.3 to 811.7 g/kg) in the affected sour lime. This study demonstrates for the first time that sour lime are a favourable substrate for aflatoxin B₁ and cyclopiazonic acid production by A. flavus isolates. This is of great concern to the health of consumers.     

Manipulating the Length of the <Emphasis Type="Italic">b</Emphasis> Subunit F<Subscript>1</Subscript> Binding Domain in F<Subscript>1</Subscript>F<Subscript>0</Subscript> ATP Synthase from <Emphasis Type="Italic">Escherichia coli</Emphasis>

The peripheral stalk of F₁F₀ ATP synthase is composed of a parallel homodimer of b subunits that extends across the cytoplasmic membrane in F₀ to the top of the F₁ sector. The stalk serves as the stator necessary for holding F₁ against movement of the rotor. A series of insertions and deletions have been engineered into the hydrophilic domain that interacts with F₁. Only the hydrophobic segment from {val-121} to {ala-132} and the extreme carboxyl terminus proved to be highly sensitive to mutation. Deletions in either site apparently abolished enzyme function as a result of defects is assembly of the F₁F₀ complex. Other mutations manipulating the length of the sequence between these two areas had only limited effects on enzyme function. Expression of a b subunit with insertions with as few as two amino acids into the hydrophobic segment also resulted in loss of F₁F₀ ATP synthase. However, a fully defective b subunit with seven additional amino acids could be stabilized in a heterodimeric peripheral stalk within a functional F₁F₀ complex by a normal b subunit.     

Inhibitory role of TGIF in the As<Subscript>2</Subscript>O<Subscript>3</Subscript>-regulated p21<Superscript><Emphasis Type="Italic">WAF1/CIP1</Emphasis></Superscript> expression

Although arsenic is an infamous carcinogen, it has been effectively used to treat acute promyelocytic leukemia, and can induce cell cycle arrest or apoptosis in human solid tumors. Previously, we had demonstrated that opposing effects of ERK1/2 and JNK on p21 expression in response to arsenic trioxide (As₂O₃) are mediated through the Sp1 responsive elements of the p21 promoter in A431 cells. Presently, we demonstrate that Sp1, and c-Jun functionally cooperate to activate p21 promoter expression through Sp1 binding sites (−84/−64) by using DNA affinity binding, chromatin immunoprecipitation, and promoter assays. Surprisingly, As₂O₃-induced c-Jun(Ser63/73) phosphorylation can recruit TGIF/HDAC1 to the Sp1 binding sites and then suppress p21 promoter activation. We suggest that, after As₂O₃ treatment, the N-terminal domain of c-Jun phosphorylation by JNK recruits TGIF/HDAC1 to the Sp1 sites and then represses p21 expression. That is, TGIF is involved in As₂O₃-inhibited p21 expression, and then blocks the cell cycle arrest.     

Vitamin B<Subscript>12</Subscript>-independent strains of <Emphasis Type="Italic">Methylophaga marina</Emphasis> isolated from Red Sea algae

Two strains (KM3 and KM5) of halophilic methylobacteria isolated from Red Sea algae do not require vitamin B₁₂ for growth and can use methanol, methylamine, dimethylamine, trimethylamine, dimethyl sulfide, and fructose as sources of carbon and energy. The cells of these strains are gram-negative motile monotrichous (strain KM3) or peritrichous (strain KM5) rods. The strains are strictly aerobic and require Na⁺ ions but not growth factors. They are oxidase-and catalase-positive and reduce nitrates to nitrites. Both strains can grow in a temperature range of 4 to 37°C (with optimal growth at 29–34°C), at pH between 5.5 and 8.5 (with optimal growth at pH 7.5–8.0), and in a range of salt concentrations between 0.5 and 15% NaCl (with optimal growth at 5–9% NaCl). The phospholipids of these strains are dominated by phosphatidylethanolamine and phosphatidylglycerol and also include phosphatidylcholine, phosphatidylserine, and cardiolipin. The dominant fatty acids are C_16:1ω7c and C_16:0. The major ubiquinone is Q₈. The cells accumulate ectoin, glutamate, and sucrose as intracellular osmoprotectants. The strains implement the 2-keto-3-deoxy-6-phosphogluconate-dependent variant of the ribulose monophosphate pathway. The G+C content of the DNA is 44.4–44.7 mol%. Analysis of the 16S rRNA genes showed that both strains belong to Gammaproteobacteria and have a high degree of homology (99.4%) to Methylophaga marina ATCC 35842^T. Based on the data of polyphasic taxonomy, isolates KM3 and KM5 are identified as new strains M. marina KM3 (VKM B-2386) and M. marina KM5 (VKM B-2387). The ability of these strains to produce auxins (indole-3-acetic acid) suggests their metabolic association with marine algae.     

Intergeneric conjugal transfer of plasmid DNA from<Emphasis Type="Italic"> Escherichia coli</Emphasis> to<Emphasis Type="Italic"> Kitasatospora setae</Emphasis>, a bafilomycin B<Subscript>1</Subscript> producer

An effective transformation procedure for Kitasatospora setae was established based on transconjugation from Escherichia coli ET12567 (pUZ8002) using a C31-derived integration vector, pSET152, containing oriT and attP fragments. While no transconjugation was observed under the standard transconjugation conditions for Streptomyces species, sufficient transconjugation (>1×10^-6) was achieved on ISP4 medium containing 30 mM MgCl₂ using a 25- to 125-fold excess of E. coli donor cells. In addition, the sequence and location of the chromosomal integration site attB of K. setae was identified for the first time in genera of non-Streptomyces actinomycetes. K. setae contains a single C31 attB site. Similar to the case of Streptomyces species, the attB site of K. setae is present within an ORF encoding a pirin-homolog, but the K. setae-attB sequence deviates slightly from the consensus sequence of Streptomyces attB sequences.     

Occurrence of fumonisin B1 in maize and poultry feeds in Haryana,India

A total of 100 maize and 50 poultry feed samples collected in 1998 at random from nine and eight districts of Haryana, respectively, were analysed for fumonisin B₁. The samples were collected from poultry farms, feed manufacturers and markets. Ninety one (91%) maize samples and forty two (84%) poultry feed samples were found to contain fumonisin B₁. Fumonisin B₁ contamination in the maize samples ranged from 0.1–87.0 ppm. Whereas the poultry feed samples contained fumonisin B₁ in the range of 0.02–28.0 ppm. It indicated widespread prevalence of fumonisin B₁ in maize and poultry feeds in different areas of Haryana. This revised version was published online in June 2006 with corrections to the Cover Date.     

DNA Damage in Astrocytes Exposed to Fumonisin B1

Fumonisins are a group of toxic metabolites mainly produced by Fusarium moniliforme and Fusarium proliferatum, fungi that commonly occur on corn throughout the world. Fumonisin B₁ (FB₁), structurally resembling sphingoid bases, is an inhibitor of ceramide synthase, a key enzyme involved in de novo sphingolipid biosynthesis and in the reacylation of free sphingoid bases derived from sphingolipid turnover. This inhibitory effect leads to accumulation of free sphinganine (SA) and sphingosine (SO), inducing cell death. However, little is known on the down stream effectors activated by these sphingolipids in the cell death signaling pathway. We exposed rat astrocytes to FB₁ with the aim of evaluating the involvement of oxygen free radicals and of some other biochemical pathways such as caspase-3 activity and DNA damage. Our results indicate that FB₁ treatment (48, 72 h and 6 days in vitro, DIV, and 10, 50, 100 M) does not affect cell viability. Conversely, after 72 h of treatment, FB₁ (50 and 100 M) induced DNA damage and an enhancement of caspase-3 activity compared to controls. In addition, FB₁ increased the expression of HSP70 at 10 and 50 M at 48, 72 h, and 6 DIV of treatment. We conclude that DNA damage of apoptotic type in rat astrocytes is caused by FB₁ and that the genotoxic potential of FB₁ has probably been underestimated and should be reconsidered.     

Low-solubility glycerol dehydratase,a chimeric enzyme of coenzyme B<Subscript>12</Subscript>-dependent glycerol and diol dehydratases

Coenzyme B₁₂-dependent diol and glycerol dehydratases are isofunctional enzymes, which catalyze dehydration of 1, 2-diols to produce corresponding aldehydes. Although the two types of dehydratases have high sequence homology, glycerol dehydratase is a soluble cytosolic enzyme, whereas diol dehydratase is a low-solubility enzyme associated with carboxysome-like polyhedral organelles. Since both the N-terminal 20 and 16 amino acid residues of the β and γ subunits, respectively, are indispensable for the low solubility of diol dehydratase, we constructed glycerol dehydratase-based chimeric enzymes which carried N-terminal portions of the β and γ subunits of diol dehydratase in the corresponding subunits of glycerol dehydratase. Addition of the diol dehydratase-specific N-terminal 34 and 33 amino acid residues of the β and γ subunits, respectively, was not enough to lower the solubility of glycerol dehydratase. A chimeric enzyme which carries the low homology region (residues 35–60) of the diol dehydratase β subunit in addition to the diol dehydratase-specific extra-regions of β and γ subunits showed low solubility comparable to diol dehydratase, although its hydropathy plot does not show any prominent hydrophobic peaks in these regions. It was thus concluded that short N-terminal sequences are sufficient to change the solubility of the enzyme.     

Leukotriene B<Subscript>4</Subscript> regulates proliferation and differentiation of cultured rat myoblasts via the BLT1 pathway

Skeletal muscle regeneration is a highly orchestrated process initiated by activation of adult muscle satellite cells. Upon muscle injury, the inflammatory process is always accompanied by muscle regeneration. Leukotriene B₄ is one of the essential inflammatory mediators. We isolated and cultured primary satellite cells. RT-PCR showed that myoblasts expressed mRNA for LTB₄ receptors BLT1 and BLT2, and LTB4 promoted myoblast proliferation and fusion. Quantitative real-time PCR and immunoblotting showed that LTB₄ treatment expedited the expression process of differentiation markers MyoD and M-cadherin. U-75302, a specific BLT1 inhibitor, but not LY2552833, a specific BLT2 inhibitor, blocked proliferation and differentiation of myoblasts induced by LTB₄, which implies the involvement of the BLT1 pathway. Overall, the data suggest that LTB₄ contributes to muscle regeneration by accelerating proliferation and differentiation of satellite cells. These authors contributed equally to this work.     

The <Emphasis Type="Italic">b</Emphasis><Subscript>arg36</Subscript> contributes to efficient coupling in F<Subscript>1</Subscript>F<Subscript>O</Subscript> ATP synthase in <Emphasis Type="Italic">Escherichia coli</Emphasis>

In Escherichia coli, the F₁F_O ATP synthase b subunits house a conserved arginine in the tether domain at position 36 where the subunit emerges from the membrane. Previous experiments showed that substitution of isoleucine or glutamate result in a loss of enzyme activity. Double mutants have been constructed in an attempt to achieve an intragenic suppressor of the b _arg36→ile and the b _arg36→glu mutations. The b _arg36→ile mutation could not be suppressed. In contrast, the phenotypic defect resulting from the b _arg36→glu mutation was largely suppressed in the b _{arg36→glu,glu39→arg} double mutant. E. coli expressing the b _{arg36→glu,glu39→arg} subunit grew well on succinate-based medium. F₁F_O ATP synthase complexes were more efficiently assembled and ATP driven proton pumping activity was improved. The evidence suggests that efficient coupling in F₁F_O ATP synthase is dependent upon a basic amino acid located at the base of the peripheral stalk.     

Occurrence of fumonisins (B<Subscript>1</Subscript>, B<Subscript>2</Subscript>, B<Subscript>3</Subscript>) in maize-based food and feed samples from Indonesia

A survey to evaluate the contamination level of total fumonisins in maize-based foodstuffs, maize and feed from Indonesia is described. The analyses were carried out by enzyme-linked immunosorbent assay (ELISA). Samples were collected from local retail stores around Yogyakarta, Indonesia between February and May 2001. The 101 samples were classified into six categories, i.e. industrially-produced food (n=24), products of small food manufacturers (n=17), maize flour (n=4), maize for food (n=9), maize for feed (n17), and formulated feed (n30). Control of the method showed that the detection limit was 8.7 μg/kg and repeatability is shown by relative standard deviation (RSD) of analyses of contaminated maize (n=5) of 10 %. Results of analyses indicate that 80 samples analysed were contaminated over a large range from 10.0-3307 pg/kg, and the concentration of fumonisins depended on the type of sample. Of four samples of maize flour, none were contaminated (below detection limit). Of 24 samples of industrially produced food, 14 were contaminated in the range 22.8 - 105 μg/kg and 18 of 19 food samples from small manufacturers were contaminated ranging from 12.9 to 234 μg/kg. The highest contamination was observed in maize samples: six of ten samples of maize for food were contaminated between 68.0 - 2471 μg/kg and 16 of 17 samples for feed contained fumonisins over a large range from 17.6 to 3306 μg/kg.