Toxicological effects of fumonisin B1 in combination with other Fusarium toxins.

Fusarium is a fungal genus spread worldwide commonly associated to the production of several mycotoxins, where fumonisins (FBs) are of major importance due to its prevalence. Since mycotoxins have been reported to cause deleterious effects on mammalians, including carcinogenic, neurotoxic, estrogenic, and immune-suppressive, many countries had established regulations on the tolerated concentrations of such substances in foods and animal feed. Even though many mycotoxins - especially fusariotoxins - are concomitantly found in a single matrix, there is no regulation on co-occurrence levels. This is possibly a result of the lack of data in the literature on the toxicological interactions between different mycotoxins. Considering this, it is of utmost importance to gather what is currently known about the combination of FBs, considered to be the most ubiquitous mycotoxins, with other frequently reported fusariotoxins, such as zearalenone (ZEA), deoxynivalenol (DON), nivalenol (NIV), T-2 toxin (T-2), and other emerging mycotoxins. This paper gives an overview about the toxic effects of fusariotoxins individually and combined to FB1, also gathering the mechanisms and probable interactions between them. This important information may help to develop regulations covering multi-mycotoxins contamination, a growing concern of current days.

Creatine kinase and ATPase activities in piglets fed a fungal mycotoxin co-contaminated diet: Consequences in the pathogenesis of subclinical intoxication.

Creatine kinase (CK) activity, through the creatine-kinase-phosphocreatine (CK/PCr) system, provides a temporal and spatial energy buffer to maintain cellular energetic homeostasis, being responsible to provide adenosine triphosphate (ATP) to the proper function of ATPases enzymes, such as the sodium-potassium (Na+, K+-ATPase) and hydrogen (H+-ATPase) pumps. Thus, the aim of this study was to evaluate the involvement of CK/PCr system in the impairment of energetic homeostasis in piglets fed with a diet co-contaminated with mycotoxins, as well as the effects on ATPases enzymes. Animals were randomly divided in two groups (eight repetitions with two animals each): CON (basal diet) and MYC (mycotoxin diet; 9300 µg/kg of aflatoxins and 8000 µg/kg of fumonisins) which were feed during 15 days. Piglets that received a diet containing 300 µg/kg of aflatoxins and 8000 µg/kg of fumonisins (MYC group) presented lower body weight on days 10 and 15 of experiment when compared to control (CON group). Serum CK activity was lower on days 5, 10 and 15 of experiment in the MYC group. The same occurred for serum Na+, K+-ATPase and H+-ATPase activities on days 10 and 15 when compared to CON group. Moreover, serum calcium levels were superior on day 15 of experiment in the MYC group, while serum potassium and sodium levels were lower in comparison to CON group. Based on these evidences, a diet co-contaminated by aflatoxins and fumonisins inhibits serum CK activity, impairing the energetic homeostasis. This inhibition alters the activities of ATPases (Na+, K+-ATPase and H+-ATPase), contributing to the imbalance of Na+, K+ and Ca+ ionic levels. In summary, the cascade of alterations contributes directly to disease pathogenesis of piglets intoxicated by mycotoxins.

Fumonisins B1 and B2 in the corn-milling process and corn-based products, and evaluation of estimated daily intake.

The distribution of fumonisins (FBs: FB1 and FB2) in the corn-milling process and in corn-based products, as well as daily intake estimates for the Brazilian population were evaluated. Among corn fractions samples, corn meal had the highest mean concentration of FB1 (1305 µg kg(-1)) and FB2 (651 µg kg(-1)) and a distribution factors of 452% and 256% in relation to corn grain, respectively. On the other hand, the distribution factor of FB1 and FB2 in corn flour was found to be 144% and 88% respectively, which demonstrates that fumonisins in this fraction were reduced compared with corn grain. As a result, almost half the corn meal samples (47%) would be non-compliant with future Brazilian regulation (2017) for fumonisins. However, corn-based products, such as corn flakes and popcorn, were in compliance with the regulation. The average probable daily intake and maximum probable daily intake of fumonisins estimated for the Santa Catarina state (Brazil) population were below the provisional maximum tolerable daily intake of 2 µg kg(-1) body weight day(-1) for all corn samples. Despite this, the adoption of practices to control the occurrence of fumonisins should be applied to the corn-milling fractions that may contain a higher concentration of this toxin, such as corn meal, often used for animal feed in Brazil.

Urinary fumonisin B1 and estimated fumonisin intake in women from high- and low-exposure communities in Guatemala.

SCOPE: Fumonisin (FB) intake can be high when maize is a dietary staple. We determined (i) urinary FB (UFB) in women consuming maize in high- and low-exposure communities in Guatemala, (ii) the FB levels in maize, (iii) the relationship between UFB and FB intake, and (iv) the relative excretion of UFB1 , UFB2 , and UFB3 . METHODS AND RESULTS: Urine and maize were analyzed for FB for 1 year in three departments. Maize consumption was estimated by an interview questionnaire. Fumonisin B1 , B2 , and B3 (FB1 , FB2 and FB3 ), were detected in 100% of maize samples. FB1 in maize and urine was significantly higher in Jutiapa compared to Chimaltenango or Escuintla. The FB intake paralleled UFB1 in a dose-dependent manner but UFB1 was present in much higher levels than UFB2 or UFB3 compared to maize. CONCLUSION: In Jutiapa, agroecological conditions favored FB production. UFB1 mirrored the estimated FB intake. UFB1 > 0.1 ng/mL resulted in a dose-dependent increase in the risk of exceeding FB intake of 2 µg/kg b.w./day compared to women with no detectable UFB1 . More than 50% exceeded 2 µg/kg b.w./day when UFB1 was >0.5 ng/mL. UFB2 and UFB3 were rarely detected confirming that FB1 is either absorbed better or preferentially excreted in urine.

Lipid metabolism of commercial layers fed diets containing aflatoxin, fumonisin, and a binder.

Aflatoxins (AF) and fumonisins (FU) are a major problem faced by poultry farmers, leading to huge economic losses. This experiment was conducted to determine the effects of AF (1 mg/kg of feed) and FU (25 mg/kg of feed), singly or in combination, on the lipid metabolism in commercial layers and investigate the efficacy of a commercial binder (2 kg/t of feed) on reducing the toxic effects of these mycotoxins. A total of 168 Hisex Brown layer hens, 37 wk of age, were randomized into a 3 × 2 + 1 factorial arrangement (3 diets with no binder containing AF, FU, and AF+FU; 3 diets with binder containing AF, FU, and AF+FU; and a control diet with no mycotoxins and binders), totaling 7 treatments. The hens contaminated with AF showed the characteristic effects of aflatoxicosis, such as a yellow liver, resulting from the accumulation of liver fat, lower values of plasma very low-density lipoprotein and triglycerides, and higher relative weight of the kidneys and liver. Hepatotoxic and nephrotoxic effects of FU were not observed in this study. On the other hand, the FU caused a reduction in small intestine length and an increase in abdominal fat deposition. The glucan-based binder prevented some of the deleterious effects of these mycotoxins, particularly the effects of AF on hepatic lipid metabolism, kidney relative weight, and FU in the small intestine.

Individual and combined effects of Salmonella typhimurium lipopolysaccharide and fumonisin B1 in broiler chickens.

The aim of this research was to evaluate the individual and combined effects of Salmonella typhimurium lipopolysaccharide (sLPS) and fumonisin B(1) (FB) on performance, relative weight of liver, biological parameters, and histological evaluation of several tissues from four hundred thirty-two 1-d-old male broiler chickens divided into 9 treatments according to the dose of FB (0, 100, or 200 mg/kg, from d 1 to d 28) and sLPS (0, 250, or 500 µg/application per bird, every other day, from d 15 to 27) administered. At the end of the experiment (28 d), significant effects caused by sLPS, FB, and the interaction of sLPS × FB were observed on several parameters. Histopathological evaluations showed significant lesions in liver and kidney caused by sLPS, FB, and their association. According to these results, both sLPS and FB (isolated or in association) cause significant effects on performance and biological parameters of broilers at 28 d of age.

Toxicokinetics and toxicological effects of single oral dose of fumonisin B1 containing Fusarium verticillioides culture material in weaned piglets.

Toxicokinetics and the toxicological effects of culture material containing fumonisin B(1) (FB(1)) were studied in male weaned piglets by clinical, pathological, biochemical and sphingolipid analyses. The animals received a single oral dose of 5 mg FB(1)/kg of body weight, obtained from Fusarium verticillioides culture material. FB(1) was detected by HPLC in plasma collected at 1-h intervals up to 6h and at 12-h intervals up to 96 h. FB(1) eliminated in feces and urine was quantified over a 96-h period and in liver samples collected 96 h post-intoxication. Blood samples were obtained at the beginning and end of the experiment to determine serum enzyme activity, total bilirubin, cholesterol, sphinganine (Sa), sphingosine (So) and the Sa/So ratio. FB(1) was detected in plasma between 30 min and 36 h after administration. The highest concentration of FB(1) was observed after 2 h, with a mean concentration of 282 microg/ml. Only 0.93% of the total FB(1) was detected in urine between 75 min and 41 h after administration, the highest mean concentration (561 microg/ml) was observed during the interval after 8 at 24 h. Approximately 76.5% of FB(1) was detected in feces eliminated between 8 and 84 h after administration, with the highest levels observed between 8 and 24 h. Considering the biochemical parameters, a significant increase only occurred in cholesterol, alkaline phosphatase and aspartate aminotransferase activities. In plasma and urine, the highest Sa and Sa/So ratios were obtained at 12 and 48 h, respectively.

Evaluation of sphingolipids in Wistar rats treated to prolonged and single oral doses of fumonisin b1.

The objective of the present study was to evaluate sphingolipid levels (sphingosine-So and sphinganine-Sa) and to compare the Sa/So ratio in liver, serum and urine of Wistar rats after prolonged administration (21 days) of fumonisin B(1) (FB(1)). In parallel, the kinetics of sphingolipid elimination in urine was studied in animals receiving a single dose of FB(1). Prolonged exposure to FB(1) caused an increase in Sa levels in urine, serum and liver. The most marked effect on sphingolipid biosynthesis was observed in animals treated with the highest dose of FB(1). Animals receiving a single dose of FB(1) presented variations in Sa and So levels and in the Sa/So ratio.

Acute toxicity of a single gavage dose of fumonisin B1 in rabbits.

The aim of this study was to determine the clinical, pathological and mycotoxicological effects of oral administration of fumonisin B1 (FB1) in rabbits. Eighteen rabbits were randomly assigned to two experimental groups: control group, 0 mg FB1; fumonisin group, 31.5 mg FB1/kg body weight, corresponding to about 630 mg FB1/kg diet. Fumonisin administered as a single oral dose to rabbits resulted in acute toxicity, significantly interfering with body and liver weight. Serum biochemical analysis revealed a significant increase of total protein, alkaline phosphatase (AP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT), urea and creatinine in the group receiving FB1 compared to control animals, a finding characterizing hepatic and renal injury in this group. Urinary protein concentrations were markedly elevated at 12, 24, 48 and 72 h after dosing, although visible pathological abnormalities were not observed, probably because of rapid repair of the damage. FB1 was detected in feces, with a maximum concentration at 24 h after administration, indicating that the enterohepatic circulation is important in rabbits. FB1 concentrations found in urine were low, with peak elimination at 12 h after intoxication. The highest FB1 concentrations were observed in feces compared to urine and liver, demonstrating that feces are the main routes of excretion.

Effects of oral administration of aflatoxin B1 and fumonisin B1 in rabbits (Oryctolagus cuniculus).

The effects of prolonged oral administration (21 days) of fumonisin B(1) (FB(1)) and aflatoxin B(1) (AFB(1)) were studied in male New Zealand rabbits by clinical, pathological, biochemical and sphingolipid analyses. Twenty-four animals were randomly divided into the following four experimental groups: (A) 0 mg FB(1)+0 microg AFB(1)/(kg body weight(bw)day) (control); (B) 0 mg FB(1)+30 microg AFB(1)/(kg bw day); (C) 1.5 mg FB(1)/(kg bw day)+30 microg AFB(1)/(kg bw day); (D) 1.5 mg FB(1)/(kg bw day)+0 microg AFB(1). Animals from group B and principally from group C presented clinical signs of intoxication. Rabbits from group C presented a lower body weight gain than controls. Differences were observed between intoxicated rabbits and controls with respect to absolute and relative liver and kidney weight, hepatic function, serum urea and creatinine levels and Sa/So ratio. The most frequent hepatic and renal injuries were vacuolar degeneration of the liver and kidney as shown by the histopathological and serum biochemical results. Combined administration of AFB(1) and FB(1) resulted in synergistic toxic effects both in the liver and in the kidney, but hepatic injuries were more marked.

Effects of prolonged oral administration of aflatoxin B1 and fumonisin B1 in broiler chickens.

The effects of prolonged oral administration of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) mycotoxins were evaluated in broiler chickens from 21 to 42 d of age. A total of 192 birds were housed in experimental batteries and assigned to 32 cages, 6 birds per cage. The following treatments were applied: 1) 0 mycotoxins (control), 2) 10 mg of FB1, 3) 50 microg of AFB1, 4) 50 microg of AFB1 + 10 mg of FB1, 5) 350 microg of AFB1, 6) 350 microg of AFB1 + 10 mg of FB1, 7) 2,450 microg of AFB1, 8) 2,450 microg of AFB1 + 10 mg of FB1/kg of feed. Each treatment consisted of 4 replicates of 6 birds each. At the end of the trial, blood samples from 12 birds per treatment were collected, and the birds were necropsied. Compared with controls, the percentage of heterophils was lower (P < 0.05) in birds from groups receiving 50 microg of AFB1/kg + 10 mg of FB1/ kg and 2450 microg of AFB1/kg alone or in combination with FB1. A higher percentage of lymphocytes (P < 0.05) was observed in birds fed 50 microg of AFB1/kg + 10 mg of FB1/ kg, 350 microg of AFB1/kg, and 2,450 microg of AFB1/kg. A decrease in plasma albumin was observed only in birds fed 2,450 microg of AFB1/kg + 10 mg of FB1/kg. The liver of AFB1-treated birds had focal areas of necrosis and inflammatory infiltrates. In birds fed rations containing only 10 mg of FB1/kg, bile duct hyperplasia with fibrosis and a mononuclear infiltrate accompanied by trabecular derangement were observed. In contrast, in treatments in which FB1 was administered in combination, hepatic vacuolar degeneration was observed, and renal tissue presented corpuscles with increased cellular agglomeration, characterizing glomerulonephritis, and a clearly visible tubular epithelium with areas of degeneration and necrosis. The FB1 residues were detected in liver and in excreta of all FB1-treated groups, at levels that ranged from 0.013 to 0.051 mg/kg and from 1.19 to 2.79 mg/kg, respectively. Results indicated that FB1 and AFB1, singly or in combination at the levels evaluated, do not change markedly the hematological and serological parameters of broiler chickens, but may cause relevant lesions in liver and in kidneys.

Effects of prolonged administration of aflatoxin B1 and fumonisin B1 in laying Japanese quail.

In the present study, 288 8-wk-old Japanese quail were randomly distributed into 6 experimental groups (48 birds per group) and fed the following diets for 140 d: 1) 0 (control); 2) 10 mg of fumonisin B1 (FB1); 3) 50 microg of aflatoxin B1 (AFB1); 4) 50 microg of AFB1 + 10 mg of FB1; 5) 200 microg of AFB1; and 6) 200 microg of AFB1 + 10 mg of FB1/kg of feed. Each treatment consisted of 4 replicates of 12 quail. Egg production and individual egg weight were checked daily. Feed intake and feed conversion were determined weekly. Results showed that by the end of the fifth cycle, average egg weight was lower (P < 0.05) in groups fed 10 mg of FB1/kg, 50 microg of AFB1/kg, 200 microg of AFB1/kg, and 10 mg of FB1 + 50 microg of AFB1/kg of feed. Egg production decreased (P < 0.05) in birds fed 10 mg of FB1/kg by the third, fourth, and fifth cycles. Feed intake was lower (P < 0.05) in birds fed 10 mg of FB1/kg by the fourth and fifth cycles, and in birds fed 50 and 200 microg of AFB1/kg in the fifth cycle. Birds fed 10 mg of FB1 + 50 microg of AFB1/kg consumed less feed (P < 0.05) in the first, second, and fifth cycles. Results indicated that prolonged administration of FB1 and AFB1, singly or in combination at the levels evaluated, may cause economic losses to quail egg producers.

Comparison of urinary sphingolipids in human populations with high and low maize consumption as a possible biomarker of fumonisin dietary exposure.

The increased sphinganine/sphingosine (SA/SO) ratio has previously been shown as a biomarker of fumonisin exposure in experimental animals and has been proposed as a tool to assess human exposure to fumonisin mainly occurring through the dietary consumption of fumonisin contaminated maize-based foods. Sphinganine and sphingosine were measured in urines of humans resident in two areas of North Argentina and South Brazil with high maize consumption and compared with urine samples collected in areas with very low or no maize consumption, such as Central Argentina and Southern Italy. The pattern of SA/SO values in the two groups with no maize consumption (assumed as controls) was similar, with all SA/SO values lower than one. Mean SA/SO ratio was 1.27 in urine of subjects with high maize consumption (n = 123) and 0.36 in controls (n = 66) and the difference was statistically significant (p<0.001). The mean fumonisin level in maize samples collected in North Argentina and South Brazil was 0.35 mg kg(-1) (n = 40). Although a similar maize and fumonisin intake was recorded for the two groups of populations, the mean SA/SO ratio in South Brazil (1.57) was significantly higher (p<0.05) than that of North Argentina (0.69). These data suggest that the higher SA/SO values observed in South Brazil cannot be associated with high fumonisin exposure and further studies are necessary to provide convincing evidence for using the SA/SO ratio as a biomarker of human fumonisin exposure.