Control of fumonisin: effects of processing.

Of about 10 billion bushels of corn that are grown each year in the United States, less than 2% is processed directly into food products, and about 18% is processed into intermediates such as high-fructose corn syrup, ethanol, and cornstarch. The vast majority of the annual crop is used domestically for animal feed (60%), and about 16% is exported. Thus, any program for controlling residues of fumonisin (FB) in food must recognize that most of the crop is grown for something other than food. Studies on the effects of wet milling on FB residues found these residues nondetectable in cornstarch, the starting material for high-fructose corn syrup and most other wet-milled food ingredients. Similar effects are noted for the dry-milling process. FB residues were nondetectable or quite low in dry flaking grits and corn flour, higher in corn germ, and highest in corn bran. Extrusion of dry-milled products reduces FB concentrations by 30-90% for mixing-type extruders and 20-50% for nonmixing extruders. Cooking and canning generally have little effect on FB content. In the masa process measurable FB is reduced following the cooking, soaking, and washing steps, with little conversion of FB to the hydrolyzed form. Sheeting, baking, and frying at commercial times and temperatures generally have no effect. In summary, all available studies on the effects of processing corn into food and food ingredients consistently demonstrate substantial reductions in measurable FB. No studies have shown a concentration in FB residues in food products or ingredients.

An overview of rodent toxicities: liver and kidney effects of fumonisins and Fusarium moniliforme.

Fumonisins are produced by Fusarium moniliforme F. verticillioides) and other Fusarium that grow on corn worldwide. They cause fatal toxicoses of horses and swine. Their effects in humans are unclear, but epidemiologic evidence suggests that consumption of fumonisin-contaminated corn contributes to human esophageal cancer in southern Africa and China. Much has been learned from rodent studies about fumonisin B1(FB1), the most common homologue. FB1 is poorly absorbed and rapidly eliminated in feces. Minor amounts are retained in liver and kidneys. Unlike other mycotoxins, fumonisins cause the same liver cancer promotion and subchronic (studies (3/4) 90 days) liver and kidney effects as (italic)F. moniliforme. FB 1 induces apoptosis of hepatocytes and of proximal tubule epithelial cells. More advanced lesions in both organs are characterized by simultaneous cell loss (apoptosis and necrosis) and proliferation (mitosis). Microscopic and other findings suggest that an imbalance between cell loss and replacement develops, a condition favorable for carcinogenesis. On the molecular level, fumonisins inhibit ceramide synthase, and disrupt sphingolipid metabolism and, theoretically, sphingolipid-mediated regulatory processes that influence apoptosis and mitosis. Liver sphingolipid effects and toxicity are correlated, and ceramide synthase inhibition occurs in liver and kidney at doses below their respective no-observed-effect levels. FB1 does not cross the placenta and is not teratogenic in vivoin rats, mice, or rabbits, but is embryotoxic at high, maternally toxic doses. These data have contributed to preliminary risk evaluation and to protocol development for carcinogenicity and chronic toxicity studies of FB1 in rats and mice.

Sphingolipid perturbations as mechanisms for fumonisin carcinogenesis.

There is a great deal of evidence that altered sphingolipid metabolism is associated with fumonisin-induced animal diseases including increased apoptotic and oncotic necrosis, and carcinogenesis in rodent liver and kidney. The biochemical consequences of fumonisin disruption of sphingolipid metabolism most likely to alter cell regulation are increased free sphingoid bases and their 1-phosphates, alterations in complex sphingolipids, and decreased ceramide (CER) biosynthesis. Because free sphingoid bases and CER can induce cell death, the fumonisin inhibition of CER synthase can inhibit cell death induced by CER but promote free sphingoid base-induced cell death. Theoretically, at any time the balance between the intracellular concentration of effectors that protect cells from apoptosis (decreased CER, increased sphingosine 1-phosphate) and those that induce apoptosis (increased CER, free sphingoid bases, altered fatty acids) will determine the cellular response. Because the balance between the rates of apoptosis and proliferation is important in tumorigenesis, cells sensitive to the proliferative effect of decreased CER and increased sphingosine 1-phosphate may be selected to survive and proliferate when free sphingoid base concentration is not growth inhibitory. Conversely, when the increase in free sphingoid bases exceeds a cell's ability to convert sphinganine/sphingosine to dihydroceramide/CER or their sphingoid base 1-phosphate, then free sphingoid bases will accumulate. In this case cells that are sensitive to sphingoid base-induced growth arrest will die and insensitive cells will survive. If the cells selected to die are normal phenotypes and the cells selected to survive are abnormal, then cancer risk will increase.

Persistence and reversibility of the elevation in free sphingoid bases induced by fumonisin inhibition of ceramide synthase.

These studies determined (1) the time course for sphingoid base elevation in the small intestines, liver, and kidney of mice following a single 25 mg/kg body weight (bw) oral dose (high dose) of fumonisin B(1) (FB(1)), (2) the minimum threshold dose of FB(1) that would prolong the elevated sphingoid base concentration in kidney following the single high dose, and (3) the importance of the balance between the rate of sphingoid base biosynthesis and degradation in the persistence of sphingoid base accumulation. Following the high dose of FB(1), there was an increase in sphinganine in intestinal cells and liver that peaked at 4 to 12 h and declined to near the control level by 48 h. In kidney, sphinganine peaked at 6-12 h but remained elevated until 72 h, approaching control levels at 96-120 h. Oral administration of 0.03 mg FB(1)/kg bw (low dose) for 5 days had no effect on the sphingoid bases in kidney. However, following an initial high dose, daily administration of the low dose prolonged the elevation in kidney sphinganine compared to mice receiving a single high dose. Thus, a single exposure to a high dose of FB(1) followed by daily exposure at low levels will prolong the elevation of sphinganine in kidney. In cultured renal cells FB(1) was rapidly eliminated, but elevated sphinganine was persistent. This persistence in renal cells was rapidly reversed in the presence of the serine palmitoyltransferase inhibitor (ISP-1), indicating that the persistence was due to differences in the rates of sphinganine biosynthesis and degradation. The in vivo persistence in kidney may be due to similar differences.

Tolerance to fumonisin toxicity in a mouse strain lacking the P75 tumor necrosis factor receptor.

Fumonisin B1 (FB1), a potent mycotoxin prevalent in corn and cereals, causes a variety of toxic effects in different mammalian species. The biochemical responses of FB1 involve inhibition of ceramide synthase leading to accumulation of free sphingoid bases and a possible involvement of tumor necrosis factor alpha (TNFalpha). To further characterize the role of TNFalpha, toxic response to FB1 was investigated in male C57BL/6J mice (WT) and a corresponding TNFalpha receptor knockout (TRK) strain, genetically modified to lack the TNFalpha1b receptor. The hepatotoxic effects of 5 daily injections of 2.25 mg/kg per day of FB1 were observed in WT but were reduced in TRK, evidenced by circulating alanine aminotransferase and aspartate aminotransferase levels and histopathological evaluation of the tissue. FB1 induced TNFalpha expression in the livers of both WT and TRK mice to a similar extent (3-4 fold over control); however, a corresponding increase of cellular NFkappaB, expected after the downstream cellular signaling of TNFalpha, was noted only in the WT. Accumulation of liver sphingosine after FB1 treatment was similar in both WT and TRK, but the FB1-induced increases in liver sphinganine and kidney sphingosine and sphinganine were lower in TRK than in WT. Results emphasized the role of TNFalpha in FB1-induced hepatotoxicity in mice and the possible relationship of sphingoid base accumulation and TNFalpha induction. Moreover, the presence of TNFalpha receptor 1b appears to be important in mediating the hepatotoxic responses of TNFalpha and FB1 in mice.

Time- and dose-response effects of the mycotoxin, fumonisin B(1) on sphingoid base elevations in precision-cut rat liver and kidney slices.

Fumonisins are mycotoxins produced on corn (Zea mays) by the common fungus Fusarium moniliforme. The fumonisins are potent inhibitors of sphingolipid biosynthesis and cause dramatic elevations in the free sphingoid base, sphinganine, both in cells in culture and in urine, blood and tissues of animals dosed with the toxins. In this study the effects of fumonisin B(1) (FB(1)) on sphingoid bases in precision-cut rat liver and kidney slices were evaluated. In liver slices exposed for 20 hr to FB(1), as little as 0.1 muM caused a 40-fold elevation in free sphinganine. Kidney slices were less responsive, and a 1 muM dose of FB(1) was required to cause a 10-fold increase in sphinganine. The amount of sphinganine in liver slices exposed to FB(1) increased in a time-dependent manner over a 72-hr period, but kidney slices exposed to the same doses of FB(1) showed a peak elevation of sphinganine after 24 hr, with a decline in the levels over the next 48 hr. Liver slices may more closely approximate the in vivo response of animals to FB, than do primary hepatocytes (in which sphinganine may be elevated > 100-fold), because the elevations in sphinganine were similar to those reported in livers of animals fed fumonisins. On the other hand, the response of kidney slices to FB(1) was substantially less than that reported in kidney tissue of FB(1)-fed rats, suggesting that kidney may accumulate toxic levels of sphingoid bases that are released from other tissues into the blood. The use of tissue slices also appears to be a useful bioassay tool for monitoring corn or other products for toxins, such as fumonisins, that elevate sphinganine levels. Crude extracts of corn screenings naturally contaminated with fumonisins produced significantly elevated sphinganine levels in liver slices, even after 50-fold dilution of the extract.

Fate of fumonisins during the production of fried tortilla chips.

The fate of fumonisin B(1) (FB(1)), a mycotoxin found in corn, during the commercial manufacture of fried tortilla chips was studied. FB(1) and hydrolyzed FB(1) (HFB(1)) concentrations in four lots of corn and in the masa, other intermediates, liquid and waste byproducts, and fried chips were determined by HPLC. FB(1) concentrations in the masa and chips were reduced significantly, up to 80% in the fried chips, compared to that in the raw corn. HFB(1) was also found in the masa and chips, but at low concentrations compared to FB(1). LC-MS analyses corroborated HPLC findings and further showed the presence of partially hydrolyzed FB(1) (PHFB(1)), which, like HFB(1), was formed during the nixtamalization (cooking/steeping the corn in alkaline water to make masa) step and found predominantly in the cooking/steeping liquid and solid waste. No significant amounts of N-(carboxymethyl)-FB(1) or N-(1-deoxy-D-fructos-1-yl)-FB(1), indicative of fumonisin-sugar adduct formation, were found. Thus, FB(1) is removed from corn and diverted into liquid and waste byproducts during the commercial production of fried tortilla chips. Nixtamalization and rinsing are the critical steps, whereas grinding, sheeting, baking, and frying the masa had little effect.

Instability of N-acetylated fumonisin B1 (FA1) and the impact on inhibition of ceramide synthase in rat liver slices.

Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium verticillioides. It inhibits ceramide synthase, which is a proposed underlying mechanism responsible for the myriad of toxic endpoints observed. We previously reported that N-acetylation of FB1 prevents ceramide synthase inhibition, but cautioned that impure preparations of FA1 can contain a contaminant with the ability to inhibit ceramide synthase. We now report that FA1 spontaneously rearranges to O-acetylated analogs. These rearrangement products are putative inhibitors of ceramide synthase. Rat liver slices exposed to impure FA1 containing O-acetylated FB1 had sphinganine/sphingosine (Sa:So) ratios of 1.15-1.64. Control slices had Sa:So ratios of 0.07-0.24. Clean-up to remove the O-acetylated FB1 yielded purified FA1, which produced Sa:So ratios in liver slices of 0.08-0.18. After storage for approximately 1 year as either a dry powder in a desiccator, or as a dried film at 4 degrees C, the purified FA1 again contained O-acetylated FB1, and was capable of ceramide synthase inhibition. FA1 was most stable in neutral solution, but in acidic solution the equilibrium shifted towards the O-acetylated forms. FA1 in solid form also rearranged, but more slowly than in acid solution. As FA1 is considerably less cytotoxic than FB1, these results provide additional support for the conclusion that a primary amino group is necessary for both ceramide synthase inhibition and toxicity.

Fusaric acid and modification of the subchronic toxicity to rats of fumonisins in F. moniliforme culture material.

Fumonisins and fusaric acid (FA) are mycotoxins produced by Fusarium moniliforme and other Fusarium which grow on corn. Fumonisins cause animal toxicities associated with F. moniliforme and, like F. monliforme, they are suspected human oesophageal carcinogens. Toxic synergism was obtained by simultaneous administration of FA and fumonisin B1 to chicks in ovo. To determine the effect of FA on in vivo toxicity of F. moniliforme culture material (CM), male rats (12 groups, n = 5/ group) were fed diets containing 0.025, 0.10 or 2.5% CM (providing dietary levels of 3.4, 18.4 or 437 ppm fumonisins, respectively) to which, at each CM level, 0, 20, 100 or 400 ppm FA were added. Additionally, an FA control group was fed 400 ppm FA only and an untreated control group was given neither FA nor culture material. Apoptosis and other effects consistent with those caused by fumonisins were present in the kidneys of animals fed 0.025% or more CM and in the livers of animals fed 2.5% CM. FA was without effect. No differences between the untreated and FA control groups were noted and no differences among the four groups (0-400 ppm FA) fed 0.025% CM, the four groups fed 0.10% CM or the four groups fed 2.5% CM were apparent. Thus, FA exerted no synergistic, additive or antagonistic effects on the subchronic in vivo toxicity of fumonisin-producing F. moniliforme.

Comparative subchronic toxicity studies of nixtamalized and water-extracted Fusarium moniliforme culture material.

Fumonisins are mycotoxins produced by Fusarium moniliforme, F. proliferatum and other Fusarium species, which are commonly found on corn, cause a variety of species-specific toxicoses, and have been linked to human oesophageal cancer in areas of southern Africa and China where corn is a dietary staple. The effect of nixtamalization, the process by which masa flour is produced by alkaline hydrolysis of corn, on the organ-specific toxicity of F. moniliforme culture material containing fumonisin B1 (FB1) was studied and the effectiveness of nixtamalization and water extraction for detoxifying culture material was compared. Male rats (n = 10/group) were fed diets containing 5% culture material equivalent weights of nixtamalized culture material (NX diet) providing 58 ppm hydrolysed FB1 but no FB1, water-extracted culture material (WE diet) providing 8 ppm FB1, or untreated culture material (CM diet) providing 71 ppm FB1 for 4 wk. An additional control group was fed a diet containing sound seed corn. Serum chemical and histopathological findings confirmed that the nixtamalized culture material was hepatotoxic and nephrotoxic. Hepatopathy was found in all rats fed the NX or CM diets. The lesions were qualitatively similar in these two groups, but were noticeably less severe in rats fed the NX diet. In contrast, only one rat fed the WE diet exhibited mild hepatopathy. Mild-to-moderate nephropathy resembling that induced by FB1 was found in all rats fed the NX, WE or CM diet. Thus, the organ-specific effects of nixtamalized culture material, containing no detectable FB1, were similar to those of the FB1-containing diet prepared from untreated culture material. Furthermore, nixtamalization was not as effective as water extraction as a detoxification method.

Fumonisin B1 and hydrolyzed fumonisin B1 (AP1) in tortillas and nixtamalized corn (Zea mays L.) from two different geographic locations in Guatemala.

Fumonisin B1 (FB1) is a common contaminant of corn worldwide and is responsible for several diseases of animals. In the preparation of tortillas, corn is treated with lime (producing nixtamal) that when heated hydrolyzes at least a portion of the FB1 to the aminopentol backbone (AP1), another known toxin. This study analyzed the amounts of FB1 and AP1 in tortillas and nixtamal from two communities in the central highlands of Guatemala where corn is a major dietary staple (Santa Maria de Jesus, Sacatepequez, and Patzicia, Chimaltenango). The amounts of FB1 and AP1 in tortillas from Santa Maria de Jesus were, respectively, 0.85 +/- 2.0 and 26.1 +/- 38.5 microg/g dry weight (mean +/- SD), and from Patzicia were 2.2 +/- 3.6 and 5.7 +/- 9.4 microg/g dry weight. Less than 6% of the tortillas from both locations contained > or = 10 microg FB1/g dry weight; whereas, 66% of the samples from Santa Maria de Jesus and 29% from Patzicia contained > or = 10 microg AP1/g dry weight. The highest amount of AP1 (185 microg/g dry weight) was found in tortillas from Santa Maria de Jesus. The highest amounts of FB1 were 6.5 and 11.6 microg/g dry weight in tortillas from Santa Maria de Jesus and Patzicia, respectively. The mean concentration of FB1 in nixtamal was significantly higher in Santa Maria de Jesus compared to Patzicia. Surprisingly, AP1 was not detected in any of the nixtamal samples. The human impact of exposure to these amounts of fumonisins is not known. However, based on findings with other animals, where corn is a dietary staple, long-term consumption of FB1 and AP1 (especially at > or = 10 microg/g of the diet) may pose a risk to human health.

Extraction, quantification, and biological availability of fumonisin B1 incorporated into the Oregon test diet and fed to rainbow trout.

The purpose of this study was (i) to determine whether pure fumonisin B1 could be incorporated into, recovered, and detected by high-pressure liquid chromatographic analysis from the semipurified Oregon test diet (OTD) used in rainbow trout feeding studies, and (ii) to determine if the incorporated fumonisin B1 was biologically available using the change in free sphingoid bases in liver, kidney, and serum as a mechanism-based biomarker. The results indicate that fumonisin is not easily quantified in the OTD. Recoveries ranged from 12 to 81% of the calculated concentrations based on the fumonisin B1 added to the OTD. However, the fumonisin B1 in the OTD was readily absorbed and biologically active as evidenced by marked increases in free sphinganine in liver, kidney, and serum. The magnitude of the increase in free sphinganine at 100 ppm in the OTD was comparable to that known to be associated with liver toxicity in rats, pigs, and ponies.

Isolation and purification of fumonisin B1 and B2 from rice culture.

Procedures are presented for growing Fusarium moniliforme MRC 826 on rice, separation of fumonisin B1 (FB1) from fumonisin B2 (FB2), purification of FB1 and preliminary procedures for purification of FB2. The mycotoxins were extracted from rice culture material (RCM) with acetonitrile-water (1:1), filtered, and the acetonitrile removed on a rotary evaporator. Preparative reverse phase liquid chromatography (LC) was used to isolate and partially purify FB1 and FB2 from the extract. The extract was applied to a C18 reverse phase cartridge. FB1 and FB2 were eluted from the cartridge by a gradient of water-acetonitrile at a flow rate of 30 mL/min. A second preparative LC procedure using 0.5% pyridine-water and two CN cartridges was used to purify FB1. The FB2 fraction was concentrated on a rotary evaporator to remove the acetonitrile. Acetonitrile was added back in sufficient quantity to redissolve the crystalline material in the fraction. An aliquot of the FB2 fraction was added to a centrifugal spinning silicic acid TLC plate. The centrifugal TLC plate was washed at 3 mL/min with a linear gradient of (A) chloroform-acetone(4:3) and (B) methanol-acetone (1:1) to elute the FB2. Gradient starting conditions were 10% methanol and ending conditions were 50% methanol. This preliminary study using the centrifugal spinning TLC showed the procedure to have the potential to be useful for purification of FB2.

Fumonisin toxicity and sphingolipid biosynthesis.

Fumonisins are inhibitors of sphinganine (sphingosine) N-acyltransferase (ceramide synthase) in vitro, and exhibit competitive-type inhibition with respect to both substrates of this enzyme (sphinganine and fatty acyl-CoA). Removal of the tricarballylic acids from fumonisin B1 reduces the potency by at least 10 fold; and fumonisin A1 (which is acetylated on the amino group) is essentially inactive. Studies with diverse types of cells (hepatocytes, neurons, kidney cells, fibroblasts, macrophages, and plant cells) have established that fumonisin B1 not only blocks the biosynthesis of complex sphingolipids; but also, causes sphinganine to accumulate. Some of the sphinganine is metabolized to the 1-phosphate and degraded to hexadecanal and ethanolamine phosphate, which is incorporated into phosphatidylethanolamine. Sphinganine is also released from cells and, because it appears in blood and urine, can be used as a biomarker for exposure. The accumulation of these bioactive compounds, as well as the depletion of complex sphingolipids, may account for the toxicity, and perhaps the carcinogenicity, of fumonisins.

Skeletal muscle metabolism in neonatal lean and obese pigs.

Nine lean and nine obese male pigs were examined at 14 d of age. The biceps femoris muscle of the obese pigs had a greater (P less than .05) percentage dry matter, protein and triglycerides than the muscle of lean pigs. The rate of oxidation of glucose to CO2 by the biceps femoris muscle was not influenced (P greater than .05) by phenotype but the rate was greater (P less than .05) when incubations were conducted in either the presence of leucine or palmitate. Glycolytic flux was lower (P less than .05) in the muscle of the obese pigs than in the muscle of lean pigs. Glycolytic flux was enhanced (P less than .05) by addition of leucine or palmitate to the incubation media. The rate of release of lactate, pyruvate, alanine, glutamine and glutamate into the media was similar between phenotypes and was not influenced by the presence of leucine or palmitate (P greater than .05). The total amount of leucine transaminated was greater (P less than .05) in the muscle of lean pigs than in the muscle of obese pigs. This was because of greater (P less than .05) rates of decarboxylation of leucine and release of alpha-ketoisocaproic acid into the media by the muscle of lean pigs when compared with the muscle of obese pigs. However, the ratio of leucine decarboxylated to alpha-ketoisocaproic acid released was similar (P greater than .05) between the two phenotypes. The ratio of palmitate oxidized (to CO2) to palmitate esterified was greater (P less than .01) in the muscle of the lean pigs than in the muscle of obese pigs. This latter finding may partially explain the greater triglyceride content of obese pig muscle. The generally lower rates of oxidation of substrates and of glycolytic flux in the biceps femoris muscle of obese pigs, when compared with lean pigs, may be associated with differences in body composition that develop during growth of lean and obese pigs.

Effect of selection for backfat thickness in swine on fetal skeletal muscle metabolism.

At 110 d of gestation, fetuses were removed from sows selected for high (obese) or for low (lean) backfat thickness. The body weights of lean (1,031 +/- 64 g) and obese (864 +/- 55 g) fetuses were not significantly different. Analysis of muscle composition and of in vitro metabolic characteristics was conducted on the biceps femoris muscle. The percentage of dry weight, protein and glycogen was greater in the muscle of obese fetuses than in the muscle of lean fetuses (P less than .01, P less than .05, and P less than .05, respectively). Percentage of muscle triglyceride was similar (P greater than .05) between the two phenotypes. The rate of glucose oxidation to CO2 tended to be greater (P less than .07) and the rate of lactate production was lower (P less than .05) in the muscle from obese fetuses than in the muscle from lean fetuses. The rates of leucine oxidation to CO2 and of palmitate oxidation to CO2 did not differ between phenotypes. The rate of alpha-ketoisocaproate release from the muscle of obese fetuses was greater (P less than .05) than from that of the lean fetuses. The rate of release of alanine and of glutamine plus glutamate did not differ between phenotypes. The rate of esterification of palmitate did not differ between phenotypes. It was concluded that abnormalities in glucose metabolism and in the partitioning of leucine between oxidation and release as the keto acid existed at 110 d of gestation in the muscle of obese fetuses. Any relation between these differences and ultimate differences in carcass composition were not evident.

Natural Occurrence of Fumonisins in Rice with Fusarium Sheath Rot Disease.

Twenty samples of rough rice (Oryza sativa) (unpolished kernels) collected during the 1995 harvest season from Arkansas (seven samples) and Texas (13 samples) were obtained from rice fields known to include plants with symptoms of Fusarium sheath rot putatively caused by Fusarium proliferatum. Samples were analyzed for fumonisin B1 (FB1) at three laboratories using three different extracting solvents by high-performance liquid chromatography (HPLC) or enzyme-linked immunosorbent assay (ELISA) methods. Forty percent of the samples were positive for FB1 at levels ≤4.3 µg/g by HPLC. The same samples contained FB1 at ≤3.6 µg/g when measured by an ELISA method. Most samples that were positive for FB1 were positive for fumonisin B2 (FB2) and fumonisin B3 (FB3) by HPLC at levels ≤1.2 µg/g. Very good agreement was obtained among the two laboratories using HPLC methods and the third using ELISA. Shelling of the unpolished rice results in hull and brown rice fractions. In a sample that contained 4.3 µg/g in whole kernels, the fumonisin level was very high in hulls (≤16.8 µg/g) and low in brown rice (≤0.9 µg/g). Milling of brown rice results in bran and white rice fractions. Fumonisins were found in bran at a level of ≤3.7 µg/g but were below the level of detection by HPLC in white rice. The presence of fumonisins (FB1, FB2, and FB3) was confirmed by fast atom bombardment/mass spectrometry. This is the first report of fumonisins in naturally contaminated rice in the United States.

Toxicity and sphinganine levels are correlated in rats fed fumonisin B(1) (FB(1)) or hydrolyzed FB(1).

Nixtamalization of Fusarium moniliforme culture material reduced, but did not eliminate, its toxicity to rats. Liver and kidney sphinganine concentration and sphinganine to sphingosine ratio of the animals fed diets containing water extracted (8 ppm fumonisin B(1) (FB(1))), nixtamalized (58 ppm hydrolyzed FB(1)), or unprocessed culture material (71 ppm FB(1)) were increased compared to those fed a diet lacking detectable fumonisins. Increases were generally correlated with the severity of hepatic and renal lesions and were highly correlated (P<0.0001) with body weight effects and serum chemical indications of hepatotoxicity. The findings are further evidence that inhibition of the enzyme ceramide synthase may be a key event in fumonisin toxigenesis.

Serine palmitoyltransferase inhibition reverses anti-proliferative effects of ceramide synthase inhibition in cultured renal cells and suppresses free sphingoid base accumulation in kidney of BALBc mice.

The purpose of this study was to determine the ability of the fungal serine palmitoyltransferase (SPT) inhibitor, myriocin, to prevent the anti-proliferative and cytotoxic effects of fumonisin B(1) in cultured pig kidney epithelial cells, LLC-PK(1). In an earlier study with LLC-PK(1) cells, ß-chloroalanine (a nonspecific SPT inhibitor) was found to inhibit the fumonisin-induced accumulation of free sphinganine by >90% but only partially reversed (50-60%) fumonisin's antiproliferative and cytotoxic effects. ß-Chloroalanine is not the ideal SPT inhibitor for this type of study because it also inhibits other pyridoxal 5'-phosphate-dependent enzymes. A potent and selective fungal SPT inhibitor (myriocin) was partially purified from liquid cultures of Isaria (=Cordyceps) sinclairii by a combination of organic extraction and column chromatography. The various fractions were bioassayed for their ability to inhibit fumonisin-induced sphinganine accumulation in LLC-PK(1) cells. The activity in partially purified material was compared to the activity of highly purified myriocin and the results expressed as myriocin equivalents. The estimated IC(50) and IC(95) for inhibition of fumonisin-induced sphinganine accumulation were approximately 1.8 and 22 nM, respectively. The IC(95) concentration of the fungal SPT inhibitor reversed the antiproliferative effects and prevented fumonisin-induced apoptosis after 48 h exposure to 50 µM fumonisin B(1). The SPT inhibitor was also effective at reducing free sphinganine in vivo. Free sphinganine concentration was reduced 60% in kidney of mice injected i.p. with SPT inhibitor plus fumonisin B(1) when compared to fumonisin B(1) alone. The ability of SPT inhibition to reduce fumonisin B(1)-induced sphinganine accumulation in vivo may be useful in the development of therapeutic agents for treatment of animals suspected to have been exposed to toxic levels of fumonisin in feeds.

Effects of fumonisin-containing culture material on pulmonary clearance in swine.

OBJECTIVE: To determine the potential effects of feeding tumonisin-containing culture material on the pulmonary clearance of blood-borne particulates and bacteria in swine. ANIMALS: 21 healthy male pigs randomly assigned to control and treated groups. PROCEDURE: Control pigs were fed a standard grower ration while culture material containing fumonisins (20 mg of hydrolyzed fumonisin B1/kg of body weight/d) was added to the feed of treated pigs for 7 days. On day 8, pigs were anesthetized with halothane and catheterized, using a sterile cut-down procedure. 18 hours after recovery from anesthesia, Monastral Blue or Pseudomonas aeruginosa was infused into the right atrium of treated and control pigs for 30 minutes and pulmonary clearance was determined. RESULTS: Pigs that were fed fumonisin-containing culture material had a significantly (P < 0.05) decreased ability to clear Monastral Blue and P aeruginosa. Ultrastructural examination of the lung indicated that uptake of copper pigment by pulmonary intravascular macrophages was decreased in treated pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Fumonisins, even when fed to pigs at sub-lethal concentrations, can inhibit pulmonary intravascular macrophages from removing particulate matter and bacteria from the circulation, thus potentially predisposing swine to infectious disease.