The Mycotoxin Fumonisin B1 Induces Metabolic and Molecular Perturbations Leading to Cell Death in Arabidopsis Cell Culture

Fumonisin B1 (FB1) is a fungal toxin produced by Fusarium spp. able to exert pleiotropic toxicity in plants. FB1 is known to be a strong inducer of the programmed cell death (PCD); however, the exact mechanism underling the plant-toxin interactions and the molecular events that lead to PCD are still unclear. Therefore, in this work we provided a comprehensive investigation of the response of the model organism Arabidopsis thaliana at the nuclear, transcriptional, and biochemical level after the treatment with FB1 at two different concentrations, namely 1 and 5µM during a time-course of 96 h. FB1 induced oxidative and nitrosative bursts and a rapid cell death in Arabidopsis cell cultures, which resembled a HR-like PCD event. Different genes involved in the regulation of PCD, antioxidant metabolism, photosynthesis, resistance, and sugar transport were upregulated, especially during the late treatment time and with higher FB1 concentration. Among the antioxidant enzymes and compounds studied, only glutathione appeared to be highly induced in both treatments, suggesting that it might be an important defense molecule induced during FB1 exposure. Collectively, these findings highlight the complexity of the defense network of A. thaliana and provide important information for the understanding of the physiological, molecular, and biochemical responses to counteract FB1-induced toxicity.

Indirect signal amplification strategy with universal probes-based lateral flow immunoassay for rapid quantitative detection of Fumonisin B1.

Fumonisin B1 (FB1) is a serious threat to the health of humans and animals. Herein, a lateral flow immunoassay based on the universal detection probes (goat anti-mouse IgG@Eu ) that...

Enzyme Linked Immunosorbent Assay for Fumonisin B1 Detection in Local Corn Seeds from Baghdad-Iraq

Fungi produce a series of toxic compounds on corn, especially Fumonisin B1 (FB1) toxin produced by Fusarium spp. and promoting cancer activity in humans and animals. This study aimed to the isolation and identification of fungi associated with local corn seeds and the detection for the presence of FB1 by using ELISA technique. Thirty samples of corn ears were collected from silos and markets in Baghdad city during the period from November 2018 to March 2019. The present study found that Fusarium was the dominant isolate among fungi in terms of the relative density 57.07%, followed by Aspergillus 31.17%, Rhizopus 3.36%, Alternaria 2.88%, Mucor 2.16%, Penicillium 1.92%, Trichothecium 0.96%, and Helminthosporium 0.48%. FB1 was detected in all samples of the silos and markets with a concentration range of 13.69 - 175.54 µg/kg. There were no significant differences in FB1concentration among samples collected from the silos and markets. Also, no relationship was found between the number of infected seeds by Fusarium spp. and FB1concentrations.

Toxic Effects of Mycotoxin Fumonisin B1 at Six Different Doses on Female BALB/c Mice

Background: Fumonisin B1 (FB1) is one of the most common mycotoxins contaminating feed and food. Although regulatory limits about fumonisins have been established in some countries, it is still very important to conduct research on lower doses of FB1 to determine the tolerance limits. The aim of this study was to investigate the effects of different concentrations of FB1, provide further evidence about the toxic doses- and exposure time-associated influence of FB1 on mice, especially low levels of FB1 for long-term exposure. Methods: Female BALB/c mice were treated intragastrically (i.g.) with fumonisin B1 (FB1) solutions (0 mg/kg body weight (BW), 0.018 mg/kg BW, 0.054 mg/kg BW, 0.162 mg/kg BW, 0.486 mg/kg BW, 1.458 mg/kg BW and 4.374 mg/kg BW) once a day for 8 weeks to obtain dose- and time-dependent effects on body and organ weights, hematology, blood chemical parameters and liver and kidney histopathology. Results: After the long-term administration of FB1, the body weights of the mice tended to decrease. Over time, FB1 first increased the relative spleen weight, then increased the relative kidney weight, and finally increased the relative liver weight. The mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), hemoglobin (HGB), white blood cells (WBC), platelets (PLT), and mean platelet volume (MPV) were significantly elevated after treatment with FB1 for 8 weeks. Moreover, exposure time-dependent responses were found for aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) level, which were coupled with hepatic histopathological findings, necroinflammation and vacuolar degeneration and detrital necrosis. Linear dose response was also found for liver histopathology, in which, even the minimum dose of FB1 exposure also caused changes. Renal alterations were moderate compared to hepatic alterations. Conclusion: In conclusion, we demonstrated the systemic toxic effects of different doses of FB1 in female BALB/c mice at different times. Our data indicated that the effects observed in this study at the lowest dose tested are discussed in relation to the currently established provisional maximum tolerable daily intake (PMTDI) for fumonisins. This study suggested that recommendations for the concentration of FB1 in animals and humans are not sufficiently protective and that regulatory doses should be modified to better protect animal and human health. The toxicity of FB1 needs more attention.

Assessment of Human Mycotoxin Exposure in Hungary by Urinary Biomarker Determination and the Uncertainties of the Exposure Calculation: A Case Study

Urinary biomarkers of mycotoxin exposure were evaluated in the case of healthy people (n = 41) and coeliac patients (n = 19) by using a multi-biomarker LC-MS/MS immunoaffinity based method capable to analyse biomarkers of nine mycotoxins, i.e., fumonisin B1 (FB1), fumonisin B2 (FB2), deoxynivalenol (DON), zearalenone (ZEN), ochratoxin A (OTA), Aflatoxin B1 (AFB1), T-2 toxin, HT-2 toxin and Nivalenol (NIV). Urinary biomarker concentrations were used to calculate the probable daily intake (PDI) of fumonisin B1, deoxynivalenol, zearalenone and ochratoxin A and compared with their tolerable daily intake (TDI). The human urinary excretion rate values reported in the literature and the 24 h excretion rate measured in piglets were used to estimate and compare the PDI values of the four mycotoxins. The highest mean biomarker concentrations were found for DON (2.30 ng/mL for healthy people and 2.68 ng/mL for coeliac patients). Mean OTA concentration was significantly higher (p < 0.001) in healthy people compared to coeliac patients. PDI calculated with piglets excretion data exceeded the TDI values by a much smaller percentage than when they were calculated from human data, especially for FB1. The uncertainties arising from the different calculations can be well perceived on the basis of these data.

The Ameliorative Effect of Lactobacillus Paracasei BEJ01 Against FB1 Induced Spermatogenesis Disturbance, Testicular Oxidative Stress and Histopathological Damage

Fumonisin B1 (FB1) was a possible carcinogenic molecule for humans as classified by the IARC on 2B group. In livestock, it was responsible for several mycotoxicosis and economic losses. Lactobacillus strains, inhabitants of a wide range of foodstuffs as well as our gastro-intestinal tract, were Generally Recognized as Safe (GRAS). Thus, the aim of this work was to evaluate the protective effect of Lactobacillus paracasei (LP) against FB1 induced reprotoxicities including testicular histopathology, sperm quality disturbance and testosterone level reduction. Pubescent mice were divided randomly into four groups as bellow: Group1: Control; Group 2: FB1 (100 µg/kg b.w); Group 3: LP (2× 109 CFU/kg b.w); Group 4: LP (2× 109 CFU/kg b.w) and FB1 (100 µg/kg b.w) to be then treated for 10 days. After the end of the treatment, animals were sacrificed; the plasma and epididyms and testicles were harvested to the reproductive system studies. Our results highlighted that LP counteracted the harmful effect generated by FB1. Indeed; it induced sperm quality reduction, oxidative stress generation and histological alterations. In conclusion, the used strain was able to prevent FB1-reproductive system damages of in balb/c mice and could be valorised as an anti-cating agent in animal FB1-contaminated diet.

HepG2 liver cells treated with fumonisin B1 in galactose supplemented media have altered expression of genes and proteins known to regulate cholesterol flux

Fumonisin B1 (FB1) contributes to mycotoxicosis in animals and has been associated with the incidence of some cancers in humans. The effect of FB1 on lipidomic profiles, sphingolipids and cholesterol levels have been demonstrated in experimental models, however, the events leading to altered cholesterol levels are unclear. This study investigates the molecular mechanisms that regulate the effect of FB1 on cholesterol homeostasis in galactose supplemented HepG2 liver cells. Galactose supplementation is a proven method utilised to circumvent the Crabtree effect exhibited by cancer cells, which forces cancer cells to activate the mitochondria. HepG2 cells were cultured in galactose supplemented media and treated with FB1 (IC50 = 25 μM) for 6 h. Cell viability was determined using the MTT assay. Metabolic status was evaluated using ATP luciferase assay, and cholesterol regulatory transcription factors (SIRT1, SREBP-1C, LXR, LDLR, PCSK9, and ABCA1) were investigated using western blotting and qPCR. FB1 in galactose supplemented HepG2 cells increased gene expression of SIRT1 (P<0.05), SREBP-1C, LXR, and LDLR; however, PCSK9 (P<0.05) was decreased. Furthermore, protein expression of SIRT1, LXR, and LDLR was elevated upon FB1 treatment, while SREBP-1C and PCSK9 were reduced. The data provides evidence that SIRT1 reduced the expression of PCSK9 and deacetylated LXR to prevent degradation of LDLR. This could result in a dysregulated cholesterol flux, which may contribute to FB1 mediated toxicity.

FumDSB Can Reduce the Toxic Effects of Fumonisin B1 by Regulating Several Brain-Gut Peptides in Both the Hypothalamus and Jejunum of Growing Pigs

Fumonisin B1 (FB1) is the most common food-borne mycotoxin produced by the Fusarium species, posing a potential threat to human and animal health. Pigs are more sensitive to FB1 ingested from feed compared to other farmed livestock. Enzymatic degradation is an ideal detoxification method that has attracted much attention. This study aimed to explore the functional characteristics of the carboxylesterase FumDSB in growing pigs from the perspective of brain–gut regulation. A total of 24 growing pigs were divided into three groups. The control group was fed a basal diet, the FB1 group was supplemented with FB1 at 5 mg/kg feed, and the FumDSB group received added FumDSB based on the diet of the FB1 group. After 35 days of animal trials, samples from the hypothalamus and jejunum were analyzed through HE staining, qRT-PCR and immunohistochemistry. The results demonstrated that the ingestion of FB1 can reduce the feed intake and weight gain of growing pigs, indicating that several appetite-related brain-gut peptides (including NPY, PYY, ghrelin and obestatin, etc.) play important roles in the anorexia response induced by FB1. After adding FumDSB as detoxifying enzymes, however, the anorexia effects of FB1 were alleviated, and the expression and distribution of the corresponding brain-gut peptides exhibited a certain degree of regulation. In conclusion, the addition of FumDSB can reduce the anorexia effects of FB1 by regulating several brain-gut peptides in both the hypothalamus and the jejunum of growing pigs.

Cunninghamella bertholletiae’s Toxins from Decomposing Cassava: Mitigation Strategy for Toxin Reduction Using Nepenthes mirabilis ‘Monkey Cup’ Digestive Fluids

A fermentation technique was utilised to assess a fungus, i.e. Cunninghamella bertholletiae/polymorpha, isolated from rotting cassava, ability to produce mycotoxins and resultant oxidation by-products of the mycotoxins using liquid chromatography–mass spectrometry (LC/MS). Thus, the mycotoxins/secondary metabolites, fumonisin B1 (FB1) and deoxynivalenol (DON) were produced while, heptadecanone, octadecanamide, octadecenal and 3-keto-deoxynivalenol (DON) were successfully identified as biodegradation by-products in the fermentation broth treated with hydrolysing ‘monkey cup’ juice from Nepenthes mirabilis. Exposure to the mycotoxins and the biodegradation by-products through consumption of contaminated produce including contact due to the cumulative presence in arable agricultural soil can be harmful to humans and animals. Therefore, this work reports on a strategy for the mitigation and reduction of mycotoxins in agricultural soil using natural plant pitcher juices from N. mirabilis’ ‘monkey cup’.