Fumonisin Toxicity Research Articles

Resveratrol alleviates fumonisin-induced intestinal cytotoxicity by modulating apoptosis, tight junction, and inflammation in IPEC-J2 porcine intestinal epithelial cells.

Fumonisins are common contaminants in the global food and environment, pose a variety of health risks to humans and animals. However, the method of mitigating fumonisin toxicity is still unclear. Resveratrol is a natural compound with antioxidant and anti-inflammatory properties. In this study, the protective effect of resveratrol against fumonisin-induced intestinal toxicity was investigated by the porcine intestinal epithelial cell line (IPEC-J2). The cells were treated with 0-40 μM fumonisin for 24 or 48 h with or without the 24 h resveratrol (15 μM) pretreatment. The data showed that resveratrol could alleviate the fumonisin B1 (FB1)-induced decrease in cell viability and amplify in membrane permeability. At the same time, it could reduce the accumulation of intracellular reactive oxygen species and increase the expression ranges of Nrf2 and downstream genes (SOD1 and NQO-1), thereby counteracting FB1-induced apoptosis. Furthermore, resveratrol was able to reduce the expression levels of inflammatory factors (TNF-α, IL-1β, and IL-6), increase the expression levels of tight junction proteins (Claudin-1, Occludin, and ZO-1), and the integrity of the IPEC-J2 monolayer. Our data also showed that resveratrol could attenuate the toxicity of the co-occurrence of three fumonisins. It is implied that resveratrol represents a promising protective approach for fumonisin, even other mycotoxins in the future. This provided a new strategy for further blocking and controlling the toxicity of fumonisin, subsequently avoiding adverse effects on the human and animal health.

Individual and Combined Cytotoxic Effects of Co-Occurring Fumonisin Family Mycotoxins on Porcine Intestinal Epithelial Cell.

Human health is seriously threatened by mycotoxin contamination, yet health risk assessments are typically based on just one mycotoxin, potentially excluding the additive or competitive interactions between co-occurring mycotoxins. In this investigation, we evaluated the individual or combined toxicological effects of three fumonisin-family B mycotoxins: fumonisin B1 (FB1), fumonisin B2 (FB2), and fumonisin B3 (FB3), by using porcine intestinal epithelial cells (IPEC). IPEC cells were exposed to various concentrations (2.5-40 μM) for 48 h, and a cell counting kit (CCK8) was used to determine cell vitality. Firstly, we discovered that they might inhibit cell viability. Additionally, the cytotoxicity of FB1 was significantly greater than that of FB2 and FB3. The results also indicated that the combinations of FB1-FB2, FB2-FB3, and FB1-FB2-FB3 showed synergistically toxicological effects at the ID10-ID50 levels and antagonistic effects at the ID75-ID90 levels. In addition, the FB1-FB3 exposure was also synergistic at the ID10-ID25 level. We also found that myriocin and resveratrol alleviated the cytotoxicity induced by fumonisin in IPEC cells. In all, this study may contribute to the determination of legal limits, the optimization of risk assessment for fumonisins in food and feed, and the development of new methods to alleviate fumonisin toxicity.

Targeted sphingolipid analysis in chickens suggests different mechanisms of fumonisin toxicity in kidney, lung, and brain.

Most of the toxic effects of fumonisins can be related to sphingolipid alteration, but there is little sphingolipidomic data in animals fed fumonisins in organs other than the liver. This study aimed to measure fumonisin B1 (FB1) in kidney, lung, and brain and determine its effects on sphingolipids. Thirty chickens divided into three groups received a diet containing 20.8mg FB1+FB2/kg for 0, 4, or 9 days. FB1 increased in kidney from 1.7 to 5.6nmol/kg and in lung from 0.5 to 1nmol/kg at 4 and 9 days, respectively. No FB1 was detected in brain. In kidney, sphinganine increased, C14-C16 ceramides decreased, whereas C18-C26 ceramides increased. Most of the changes in dihydroceramides, dihydrodeoxyceramides, deoxyceramides sphingomyelins, dihydrosphingomyelins, and hexosylceramides paralleled those on ceramides. In lung, sphinganine was unaffected by fumonisins, whereas sphinganine-1-phosphate increased. Other major changes corresponded to decreases in glycosylceramides. In brain, sphinganine was unchanged, whereas deoxysphinganine, sphingosine, C14-C20 ceramides, and C14-C20 sphingomyelins increased. These results revealed that alterations in sphingolipids in kidney were close to those measured in liver and could correspond to inhibition of ceramide synthase 5 activity. In contrast, effects of fumonisins in lung and brain cannot be explained by inhibition of ceramide synthase.

Assessment of information as regards the toxicity of fumonisins for pigs, poultry and horses.

In 2018, the EFSA Panel on Contaminants in the Food Chain (CONTAM) adopted a Scientific Opinion on the risks for animal health related to the presence of fumonisins, their modified forms and hidden forms in feed. A no observed adverse effect level (NOAEL) of 1 mg/kg feed was established for pigs. In poultry a NOAEL of 20 mg/kg feed and in horses a reference point for adverse animal health effect of 8.8 mg/kg feed was established, referred to as NOAEL. The European Commission (EC) requested EFSA to review the information regarding the toxicity of fumonisins for pigs, poultry and horses and to revise, if necessary, the established NOAELs. The EFSA CONTAM Panel considered that the term reference point (RP) for adverse animal health effects better reflects the uncertainties in the available studies. New evidence which had become available since the previous opinion allowed to revise an RP for adverse animal health effects for poultry from 20 mg/kg to 1 mg/kg feed (based on a LOAEL of 2.5 mg/kg feed for reduced intestinal crypt depth) and for horses from 8.8 to 1.0 mg/kg feed (based on case studies on equine leukoencephalomalacia (ELEM)). For pigs, the previously established NOAEL was confirmed as no further studies suitable for deriving an RP for adverse animal health effects could be identified. Based on exposure estimates performed in the previous opinion, the risk of adverse health effects of feeds containing FB1–3 was considered a concern for poultry, when taking into account the RP of 1 mg/kg feed for intestinal effects. For horses and other solipeds, the risk is considered low, although a large uncertainty associated with exposure was identified. The same conclusions apply to the sum of FB1–3 and their hidden forms.

Targeted Analysis of Sphingolipids in Turkeys Fed Fusariotoxins: First Evidence of Key Changes That Could Help Explain Their Relative Resistance to Fumonisin Toxicity.

The effects of fumonisins on sphingolipids in turkeys are unknown, except for the increased sphinganine to sphingosine ratio (Sa:So) used as a biomarker. Fumonisins fed at 20.2 mg/kg for 14 days were responsible for a 4.4 fold increase in the Sa:So ratio and a decrease of 33% and 36% in C14-C16 ceramides and C14-C16 sphingomyelins, respectively, whereas C18-C26 ceramides and C18-C26 sphingomyelins remained unaffected or were increased. Glucosyl- and lactosyl-ceramides paralleled the concentrations of ceramides. Fumonisins also increased dihydroceramides but had no effect on deoxysphinganine. A partial least squfares discriminant analysis revealed that all changes in sphingolipids were important in explaining the effect of fumonisins. Because deoxynivalenol and zearalenone are often found in feed, their effects on sphingolipids alone and in combination with fumonisins were investigated. Feeding 5.12 mg deoxynivalenol/kg reduced dihydroceramides in the liver. Zearalenone fed at 0.47 mg/kg had no effect on sphingolipids. When fusariotoxins were fed simultaneously, the effects on sphingolipids were similar to those observed in turkeys fed fumonisins alone. The concentration of fumonisin B1 in the liver of turkeys fed fumonisins was 0.06 µmol/kg. Changes in sphingolipid concentrations differed but were consistent with the IC50 of fumonisin B1 measured in mammals; these changes could explain the relative resistance of turkeys to fumonisins.

Design of automated database system for storage and management of reports on mycotoxins contaminated agricultural products in Sub-Saharan Africa

This paper discusses the idea and the design of an automated system for storage and management of mycotoxins reports for decision making. Mycotoxins are poisonous chemical compounds produced by certain fungi. Mycotoxins are fungal secondary metabolites that contaminate various feedstuffs and agricultural crops. The contamination of food by mycotoxins can occur before production, during storage, processing, transportation or marketing of the food products. High temperature, moisture content and water activity are among the factors that facilitate the production of mycotoxins in food. The five major mycotoxins produced in food and feedstuffs are Aflatoxins, ochratoxins, fumonisins, deoxynivalenol and zearalenone. In Africa, mycotoxin contamination is considered to be a major problem with implications that causes human and animal health hazards and poor economy. Aflatoxin-related hepatic diseases are reported in many African countries. Ochratoxin and fumonisin toxicity in humans and animals is widespread in Africa. The available and updated information on the incidence of mycotoxin is not collectively vivid for policy making. A complete automated system allows to monitor the statistical report of mycotoxins stored in agricultural products. This study involves analytical Service conducted on Mycotoxins such as Mold Culture and Identification and Chemical Analysis which involves microbiological Culturing; Microscopic or biochemical identification, enzyme linked Immunosorbent (ELISA), tin layer Chromatography (TLC), high Performance Liquid Chromatography (HPLC), and gas Chromatography /Mass Spectroscopy. The design and development of Mycotoxins Automated Database System (MADAS) makes provision for easy access and acknowledgment of mycotoxins in different grains, fruits, vegetables and foods in Sub-Saharan Africa. It also enhances robust data collection, management, and analysis, a secure and protected data environment, error reduction and data storage to facilitate regulatory compliance, improved maintainability, standardization, control, predictability, and traceability of data and lower costs due to automation of labor intensive tasks and elimination of redundant work.

UV-induction of photolytic and photocatalytic degradation of fumonisins in water: reaction kinetics and toxicity.

Although fumonisins are toxic and carcinogenic mold products that contaminate feed, food, and water, their photodegradation has not yet been reported. In this work, the efficiency of photolysis (UV, UV/H2O2, and UV/[Formula: see text]) and photocatalysis (TiO2 (Degussa P25/Wackherr) and ZnO) for the degradation of fumonisins in an aqueous medium were investigated. In the case of fumonisin B1 (FB1) optimal conditions in terms of pH, the initial concentrations of H2O2/[Formula: see text] for UV, UV/H2O2, and UV/[Formula: see text] treatments were investigated. The photocatalytic degradation using TiO2 Wackherr as catalyst at natural pH (about 8) proved to be the most efficient treatment for removal of FB1 and FB3. Namely, during the first 30 min of irradiation, 99% of FB1 (1.39 μM) was degraded, while FB3 (0.425 μM) was completely removed during the first 20 min of irradiation. In the case of FB2 (0.687 μM), UV/[Formula: see text] was the most efficient treatment, and complete removal occurred in the first 90 min of irradiation. All applied treatments for fumonisins removal have followed pseudo-first-order kinetics under the relevant experimental conditions. Toxicity of fumonisins and their mixtures formed during photodegradation were investigated using mammalian cell lines (BHK, H-4-II-E, Neuro-2a, and MRC-5). The BHK cell line was the most sensitive to fumonisins, especially FB2 and FB3, and its photodegradation mixtures.

Fumonisin B1 triggers carcinogenesis via HDAC/PI3K/Akt signalling pathway in human esophageal epithelial cells

Fumonisin B1 (FB1) is a contaminant that commonly present in the global environment, especially in food and feed. Epidemiologic studies have shown that esophageal cancer is associated with fumonisin toxicity. However, the molecular mechanism of FB1-induced esophageal cancer is unclear. In this research, the molecular mechanism of FB1-induced cell carcinogenesis in human esophageal epithelial cells line (HEEC) was explored. We found that FB1 (0.3125–5 μM) could promote cell proliferation, and the same phenomenon was found in a 3D cell model. FB1 could also accelerate cell migration. The expression levels of DNA damage markers were significantly increased after FB1 exposure. Meanwhile, the expression levels of cell cycle-regulated proteins and cancer-related genes were abnormal. Furthermore, FB1 significantly upregulated the histone deacetylase (HDAC) expression and activated the phosphoinositide 3 kinase (PI3K)/protein kinase B (Akt) signalling pathway. The HDAC inhibitor trichostatin A (TSA) could repressed FB1-promoted cell proliferation and abnormal phenomenon induced by FB1. Moreover, myriocin (ISP-1) could relieve FB1-enhanced HDAC expression and cell proliferation, which implied that ISP-1 may be used to block the fumonisin toxicity in the future. Our findings suggested that the HDAC/PI3K/Akt signalling pathway is a novel mechanism for FB1-induced cell carcinogenesis in HEEC and provided new ideas for the prevention and control of fumonisin toxicity, subsequently avoiding adverse effects on the ecosystem and human health.

Structure Activity Relationship for Fumonisin Phytotoxicity.

Fumonisins are mycotoxins produced by a number of species of Fusarium and Aspergillus. They are polyketides that possess a linear polyol structure with two tricarballylic acid side chains and an amine moiety. Toxicity results from their inhibition of Ceramide Synthase (CerS), which perturbs sphingolipid concentrations. The tricarballylic side chains and amine group of fumonisins are key molecular features responsible for inhibiting CerS, however their individual contributions toward overall toxicity are not fully understood. We have recently reported novel, deaminated fumonisins produced by A. niger and have identified an enzyme (AnFAO) responsible for their synthesis. Here we performed a structure/function activity assay to investigate the individual contributions of the tricarballylic acid and amine toward overall fumonisin toxicity. Lemna minor was treated at 40 μM against FB1, hydrolyzed FB1 (hFB1), deaminated FB1 (FPy1), or hydrolyzed/deaminated (hFPy1). Four end points were monitored: plant dry weight, frond surface area, lipidomics, and metabolomics. Overall, hFB1 was less toxic than FB1 and FPy1 was less toxic than hFB1. hFPy1 which lacks both the amine group and tricarballylic side chains was also less toxic than FB1 and hFB1, however it was not significantly less toxic than FPy1. Lipidomic analysis showed that FB1 treatment significantly increased levels of phosphotidylcholines, ceramides, and pheophorbide A, while significantly decreasing the levels of diacylglycerides, sulfoquinovosyl diacylglycerides, and chlorophyll. Metabolomic profiling revealed a number of significantly increased compounds that were unique to FB1 treatment including phenylalanine, asymmetric dimethylarginine (ADMA), S-methylmethionine, saccharopine, and tyrosine. Conversely, citrulline, N-acetylornithine and ornithine were significantly elevated in the presence of hFB1 but not any of the other fumonisin analogues. These data provide evidence that although removal of the tricarballylic side chains significantly reduces toxicity of fumonisins, the amine functional group is a key contributor to fumonisin toxicity in L. minor and justify future toxicity studies in mammalian systems.

Effects of dietary fumonisins on nutrients digestibility in weanling pigs

Submitted 2020-06-15 | Accepted 2020-07-01 | Available 2020-12-01 https://doi.org/10.15414/afz.2020.23.mi-fpap.23-28 The aim of the study was to ascertain the potential effects of fumonisins on nutrients’ digestibility in the gastrointestinal tract. Eighteen (n= 6 x 3) weanling pigs from the age of 35 days were administered 0, 15 or 30 mg/kg dietary fumonisins (FB1, FB2 and FB3) for a period of 21days, after an acclimatization period of 14 days. Titanium dioxide (0.5%) was added as indigestible marker to the feed and representative fecal samples were taken during a 5-day collection period in order to determine the digestibility of crude protein, crude fat, crude fiber, starch, ash, calcium, phosphorus and energy. The final body weights, cumulative feed intake and relative organ weights of all groups were were not influenced by treatment (p>0.05). Digestibility of energy, crude fiber, ash, calcium and phosphorus were significantly higher (p<0.05) in the control group relative to either 15 or 30 mg/kg treated pigs. These findings suggest that fumonisins in a dose of 15 or 30 mg/kg potentially distorts total tract nutrient digestibility in weanling pigs and thus, compromise the nutritive value of the mixed feed. Keywords: fumonisins, pigs, gastrointestinal tract, nutrients digestibility References Association of Official Analytical Chemists. (2000). AOAC International, Gaithersburg, 17. ed MD, USA. Bartók, T. et al. (2010). Detection and characterization of twenty-eight isomers of fumonisin B1 (FB1) mycotoxin in a solid rice culture infected with Fusarium verticillioides by reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight and ion trap mass spectrometry. Rapid Communications in Mass Spectrometry, 24(1), 35–42. Bouhet, S. and Oswald, I.P. (2007). The intestine as a possible target for fumonisin toxicity. Molecular Nutrition and Food Research, 51(8), 925–931. Choct M. (2009). Managing gut health through nutrition. British Poultry Science, 50(1), 9–15. Ewing, W.N. (2008). The Living Gut. 2. ed. Nottingham, UK: Nottingham University Press. Gbore, F.A., Yinusa, R.I. and Salleh, B. (2010). Evaluation of sub-chronic dietary fumonisin B1 on nutrient digestibility and growth performance of rats. African Journal of Biotechnology, 9(38), 6442–6447. Gbore, F.A. (2007). Effect of dietary fumonisin B1 on histomorphology and histopathology of organs of pubertal boars. American-Eurasian Journal of Scientific Research, 2, 75–79. Gbore, F.A. and Egbunike, G.N. (2007). Influence of dietary fumonisin B1 on nutrient utilization by growing pigs. Livestock Research for Rural Development, Volume19, Article #93. Retrieved June 5, 2020 from http://www.lrrd.org/lrrd19/7/gbor19093.htm Gelineau-van Waes, J. et al. (2009). Maternal fumonisin exposure as a risk factor for neural tube defects. Advances in Food and Nutrition Research, 56, 145–181. Grenier, B. and Applegate, T. (2013). Modulation of intestinal functions following mycotoxin ingestion: Meta-analysis of published experiments in animals. Toxins, 5(2), 396–430. Harrison, L.R. et al. (1990). Pulmonary edema and hydrothorax in swine produced by fumonisin B1, a toxic metabolite of Fusarium moniliforme. Journal of Veterinary Diagnostic Investigation, 2(3), 217–221. International Agency for Research on Cancer (IARC) (2002). Working group on the evaluation of carcinogenic risks to humans. Some traditional herbal medicines, some mycotoxins, naphthalene and styrene. IARC Monographs on the evaluation of carcinogenic risks to humans, 82, 1–556. Islami, F. et al. (2009). Oesophageal cancer in Golestan Province, a high-incidence area in northern Iran - a review. European Journal of Cancer, 45(18), 3156–3165. Merrill, A. H. et al. (2001). Sphingolipid metabolism: roles in signal transduction and disruption by fumonisins. Environmental Health Perspectives, 109(2), 283–289. Nelson, P.E., Desjardins, A.E. and Plattner, R.D. (1993). Fumonisins, mycotoxins produced by Fusarium species: biology, chemistry, and significance. Annual Review of Phytopathology, 31, 233–252. Rheeder, J.P., Marasas, W.F. and Vismer, H.F. (2002). Production of fumonisin analogs by Fusarium species. Applied and Environmental Microbiology, 68(5), 2101–2105. SPSS. (2012). SPSS for Windows version 20, SPSS: Chicago, IL, USA. Tóth, Á. et al. (2000). Effect of low doses of the mycotoxin fumonisin B1 on the body weight gain, feed intake and feed conversion rate of pigs. Agriculture, 6, 149–151. Â

Identification and Characterization of an Aspergillus niger Amine Oxidase that Detoxifies Intact Fumonisins.

Fumonisin contamination of maize damaged by Fusarium verticillioides and related species is a major problem when it is grown under warm and dry conditions. Consumption of fumonisin contaminated food and feed is harmful to both humans and livestock. Novel tools for reducing or eliminating fumonisin toxicity may be useful to the agri-feed sector to deal with this worldwide problem. Enzymes capable of catabolizing fumonisins have been identified from microorganisms that utilize fumonisins as an energy source. However, fumonisin detoxifying enzymes produced by the very species that biosynthesize the toxin have yet to be reported. Here we describe the identification and characterization of a novel amine oxidase synthesized by the fumonisin-producing fungus Aspergillus niger. We have recombinantly expressed this A. niger enzyme in E. coli and demonstrated its ability to oxidatively deaminate intact fumonisins without requiring exogenous cofactors. This enzyme, termed AnFAO (A. niger fumonisin amine oxidase), displays robust fumonisin deamination activity across a broad range of conditions, has a high native melting temperature, and can be purified to >95% homogeneity at high yield in a one-step enrichment. AnFAO is a promising tool to remediate fumonisin-contaminated feed including maize destined for ethanol production.

Evaluation of the Individual and Combined Toxicity of Fumonisin Mycotoxins in Human Gastric Epithelial Cells.

Fumonisin contaminates food and feed extensively throughout the world, causing chronic and acute toxicity in human and animals. Currently, studies on the toxicology of fumonisins mainly focus on fumonisin B1 (FB1). Considering that FB1, fumonisin B2 (FB2) and fumonisin B3 (FB3) could coexist in food and feed, a study regarding a single toxin, FB1, may not completely reflect the toxicity of fumonisin. The gastrointestinal tract is usually exposed to these dietary toxins. In our study, the human gastric epithelial cell line (GES-1) was used as in vitro model to evaluate the toxicity of fumonisin. Firstly, we found that they could cause a decrease in cell viability, and increase in membrane leakage, cell death and the induction of expression of markers for endoplasmic reticulum (ER) stress. Their toxicity potency rank is FB1 > FB2 >> FB3. The results also showed that the synergistic effect appeared in the combinations of FB1 + FB2 and FB1 + FB3. Nevertheless, the combinations of FB2 + FB3 and FB1 + FB2 + FB3 showed a synergistic effect at low concentration and an antagonistic effect at high concentration. We also found that myriocin (ISP-1) could alleviate the cytotoxicity induced by fumonisin in GES-1 cells. Finally, this study may help to determine or optimize the legal limits and risk assessment method of mycotoxins in food and feed and provide a potential method to block the fumonisin toxicity.

Challenge with fumonisin FB1 + FB2 alters profile of fatty acids in Nile tilapia fingerlings

The toxicity of fumonisin (FB) has been proposed for different fish species because it causes imbalance between the sphingoid bases and impairs several cellular processes, such as cell differentiation and growth. In this study, we tested the hypothesis that FB changes performance and modifies the fatty acid profile in the muscle of Nile tilapia fingerlings. A total of 180 fish were distributed in four treatments: control treatment (without addition of FB), 20 (FB20), 40 (FB40) and 60 (FB60) mg/kg FB) in order to measure performance, muscle yield, mineral residue, protein, total lipids and fatty acid content. The apparent daily gain was lower (Y = 0.019x2-0.124x + 0.325; R2 = 0.928) for fish fed diets containing FB compared to the control group. The negative effect on growth rates resulted in a reduction in fillet yield (P < 0.001). Crude protein was statistically lower (FB60 = 15.38%) for the higher dose than for the control group (P = 0.0003). The moisture was higher for the FB20 level, with quadratic regression effect (R2 = 0.688), the ashes had a linear regression effect (R2 = 0.811) and showed a quadratic regression effect (R2 = 0.868). Total saturated fatty acids were higher in fish fed with FB60 (37.13 ± 0.99) diet than for any other treatment. Total monounsaturated fatty acids did not vary between dietary treatments, as did total omega 6 (ω6). Total omega 3 fatty acids (ω3) were significantly higher in fish fed with FB40 (4.99 ± 0.15) and FB60 (5.20 ± 0.13) diets than for FB20 (4.36 ± 0.03) and the control treatment (4.43 ± 0.18). Total polyunsaturated fatty acids were significantly higher in fish fed with FB40 (33.70 ± 1.01) and FB60 (33.73 ± 0.29) diets. Therefore, fumonisin worsens performance in Nile tilapia fingerlings and modifies their fatty acid profile, as it increases the amount of saturated and polyunsaturated fatty acids and decreases the level of monounsaturated fatty acids.

Metabolism, Toxicity, Detoxification, Occurrence, Intake and Legislations of Fumonisins - A Review

Fumonisins are a group of mycotoxins generated by the Fusarium spp. in foods and feeds. More than 15 isomers of Fumonisin are recognized, and the B series of Fumonisins is the primary and referral isomer of Fumonisin. Fumonisin B can cause leukoencephalomalacia in rabbits and horses and porcine pulmonary edema in swine. Fumonisin B is also nephrotoxic, hepatotoxic, immunotoxic and carcinogenic. It blocks sphingolipid biosynthesis (and hinders the synthesis of ceramide) by a noticeable resemblance to sphingosine and sphinganine. This paper provides a review of the toxicity, occurrence, and mechanism of carcinogenicity, hepatotoxicity, nephrotoxicity as well as immunotoxicity of Fumonisins, which are primarily found on a variety of food and feed in Africa, America, Europe, Asia, and Oceania. In this paper, current information on contamination of feeds and foods by Fumonisins around the world is summarized. Because of economic losses induced by Fumonisins and their harmful effects on animal and human health, various procedures to detoxify infected feeds and foods have been illustrated in this review, including biological, physical, and chemical processes. Additionally, we discuss dietary intakes and maximum limits of Fumonisins in some countries.

Toxicity of Fumonisins, Deoxynivalenol, and Zearalenone Alone and in Combination in Turkeys Fed with the Maximum European Union-Tolerated Level.

Surveys of mycotoxins worldwide have shown that deoxynivalenol (DON), fumonisins (FB), and zearalenone (ZON) are the most abundant Fusarium mycotoxins (FUS) in European poultry feed, in both the level and the frequency of contamination. Previous studies reported that a combination of FUS at concentrations that individually are not toxic may negatively affect animals. However, although toxic thresholds and regulatory guidelines exist for FUS, none account for the risk of multiple contamination, which is the most frequent. The aim of this study was to compare DON, FB, and ZON toxicity, alone and in combination, in male turkey poults. Ground cultured toxigenic Fusarium strains were incorporated in corn-soybean-based feed in five experimental diets: control diet, containing no mycotoxins, DON diet (5 mg DON/kg), FB diet (20 mg FB1 + FB2/ kg), ZON diet (0.5 mg ZON/kg), and DONFBZON diet (5, 20, and 0.5 mg/kg of DON, FB1 + FB2, and ZON, respectively). Seventy male Grade Maker turkeys were reared in individual cages on mycotoxin-free diets from 0 to 55 days of age. On the 55th day, the turkeys were weighed and divided into five groups each comprising 14 birds. Each group was fed one of the five experimental diets for a period of 14 days. On the 70th day of age, feed was withheld for 8 hr, at which time a blood sample was collected, and then all the turkeys were killed, autopsied, and different tissues sampled. The weight of the different organs, analyses of performance, biochemistry, histopathology, oxidative damage, and testis toxicity revealed no significant effects attributable to FUS. Measurement of sphingolipids in the liver revealed an increase in the sphinganine to sphingosine ratio in turkeys fed diets containing FB, but had no apparent consequences in terms of toxicity. Finally, only slight differences were found in some variables and the results of this study showed no interactions between DON, FB, and ZON. Taken together, results thus suggest that the maximum tolerated levels established for individual contamination by DON, FB, and ZON can also be considered safe in turkeys fed with combinations of these FUS for a period of 14 days.

Effect of pH on the toxicity of fumonisins towards the RTL-W1 cell line and zebrafish (Danio rerio) embryos

Fumonisins are common contaminants of maize. The neutral red assay, using the RTL-W1 trout liver cell line, and the fish embryo test (FET), using zebrafish, were selected to assess the effect of pH on the cytotoxicity, acute toxicity and teratogenicity of fumonisin B1 (FB1). The results demonstrated that FB1 exerts low cytotoxicity towards RTL-W1 cells without pH adjustment (IC50 1 746 μM), and no cytotoxicity after pH-adjustment to physiological conditions. The LC50 value for FB1 in the FET (1 058 μM at 48 h) confirmed low acute toxicity. Adjusting the pH to physiological conditions reduced the acute toxicity of FB1 towards zebrafish embryos, emphasising the importance of acidity/basicity of the medium in toxicity testing. Hydrolysed FB1 was less toxic than FB1 (LC50 2 690 μM at 48 h), and neither were teratogenic towards zebrafish embryos. Results confirm that the FET may account for effects not observable in cell cultures.

Application of biomarker methods to investigate FUMzyme mediated gastrointestinal hydrolysis of fumonisins in pigs

Fumonisins are among the most prevalent mycotoxins in feedstuffs. They disrupt the sphingolipid metabolism, thereby inducing a plethora of toxic effects in livestock. Supplementation with mycotoxin-degrading enzymes is a promising strategy for the detoxification of feedstuffs in the animals’ gastrointestinal tract. Here, we evaluated the suitability of the fumonisin esterase FumD as a feed additive (FUMzyme®) for the prevention of fumonisin toxicity in pigs by using a combination of different fumonisin biomarkers (sphinganine to sphingosine (Sa/So) ratio in serum and organs, concentrations of fumonisin B1 and hydrolysed derivatives in urine and faeces). In a pre-trial, we exposed pigs to 30 mg/kg fumonisins in feed and found the minimum effective dose of FUMzyme to be 15 U/kg. In a second trial we investigated the long-term efficacy of this minimum effective FUMzyme dose to counteract toxic effects elicited by 6 weeks of exposure to 2.5 mg/kg fumonisins in a diet containing naturally contaminated maize. Supplementation of feed with the minimum effective FUMzyme dose prevented an increase in the Sa/So ratio in serum and kidneys of fumonisin exposed pigs. The Sa/So ratio in serum proved to be the most reliable biomarker. The fumonisin pattern in faeces was less suitable as biomarker for assessing the efficacy of FUMzyme due to natural gastrointestinal hydrolysis of fumonisins. Analysis of urine samples provided additional information about gastrointestinal fumonisin hydrolysis before fumonisin absorption, but was analytically challenging because of low urinary fumonisin concentrations.

Reduction of Fumonisin Toxicity by Extrusion and Nixtamalization (Alkaline Cooking)

Fumonisins are mycotoxins found in corn. They are toxic to animals and cause cancer in rodents and neural tube defects in LM/Bc mice. Reducing their concentrations in corn-based foods is therefore desirable. Chemical analysis or in vitro bioassays of food extracts might not detect toxic fumonisin reaction products that are unknown or unextractable from food matrices, thus potentially underestimating in vivo toxicity. The effectiveness of two common cooking methods, extrusion and nixtamalization (alkaline cooking), to reduce the toxicity of fumonisin-contaminated corn grits (extrusion) and whole kernel corn (nixtamalization) was shown by means of rat feeding bioassays using fumonisin-specific kidney effects as indicators of potential toxicity. A third bioassay showed that in contrast to fumonisin B1 (FB1), hydrolyzed fumonisin B1 (HFB1; formed from FB1 during nixtamalization) did not cause neural tube defects in LM/Bc mice. The findings indicate that extrusion and nixtamalization reduce the potential toxicity of FB1-contaminated corn.

Mechanistic Insight into the Biosynthesis and Detoxification of Fumonisin Mycotoxins.

Fumonisins, notably FB1, FB2, FB3, and FB4, are economically important mycotoxins produced by a number Fusarium sp. that occur on corn, rice, and sorghum as well as by Aspergillus sp. on grapes. The fumonisin scaffold is comprised of a C18 polyketide backbone functionalized with two tricarballylic esters and an alanine derived amine. These functional groups contribute to fumonisin's ability to inhibit sphingolipid biosynthesis in animals, plants, and yeasts. We report for the first time the isolation and structure elucidation of two classes of nonaminated fumonisins (FPy and FLa) produced by Aspergillus welwitschiae. Using a Lemna minor (duckweed) bioassay, these new compounds were significantly less toxic in comparison to the fumonisin B mycotoxins, providing new insight into the mechanism of fumonisin toxicity. Time course fermentations monitoring the production of FB4, FPy4, and FLa4, as well as (13)C and (15)N stable isotope incorporation, suggest a novel postbiosynthetic oxidative deamination process for fumonisins. This pathway was further supported by a feeding study with FB1, a fumonisin not produced by Aspergillus sp., which resulted in its transformation to FPy1. This study demonstrates that Aspergillus have the ability to produce enzymes that could be used for fumonisin detoxification.

Production of a phage-displayed mouse ScFv antibody against fumonisin B1 and molecular docking analysis of their interactions

Fumonisins produced by Fusarium pathogens are mycotoxins present in maize and other grains in the field as well as during storage worldwide and pose a serious threat to humans and domestic animals. Fumonisin B consists of different chemotypes, and fumonisin B1 (FB1) is the most predominant fumonisin found in food/feed commodities. Recombinant antibody can be deployed to analyze the fumonisin toxicological mechanism and develop a simple and cost-effective method for the detection of fumonisins, which is vitally important for monitoring and preventing fumonisins from entering food/feed chains. In this study, FB1 conjugated to keyhole limpet hemocyanin was used to immunize mice, from which RNA was isolated to construct a recombinant antibody library. Successive panning of the library by phage display was used to select monoclonal phage clones reactive to FB1 conjugated to bovine serum albumin. Subsequent phage ELISA and sequencing analyses revealed four different reactive scFv antibodies specific to FB1. Soluble expression and ELISA analysis showed that one scFv antibody, FBMA1, had the highest reactivity and could be purified from bacterial cells in large quantities. Surface plasmon resonance measurements further revealed that the FBMA1 scFv antibody had a binding kinetics of K D = 1.89 × 10–7 M. Molecular modeling and docking analyses suggested that the FBMA1 antibody shaped a proper cavity to embed the whole FB1 molecule and that a steady-state complex was formed relying on intermolecular forces, including hydrogen bonding, electrostatic force and hydrophobic interactions. Thus, the scFv antibody can be applied for mechanistic studies of intermolecular interactions and fumonisin toxicity, and for the development of an immunoassay for fumonisin-contaminated food/feed samples.