Fusarium species can produce several mycotoxins e.g. FB1, DON and ZEA which are frequent contaminants of cereals intended either for human or animal consumption. F. verticillioides and F. proliferatum produce FB1 whereas F. graminearum is the main producer of DON and ZEA resulting in co-occurrence. Although multimycotoxin contamination occurs very often, most studies focus on single effects. Moreover the mycotoxins’ concentrations used in the experimental diets are usually high and unlikely to occur in nature. Data on the effect of combined mycotoxins consumption in low dosages (according to EU recommendations) are scarce. In addition there are no studies assessing the genotoxicity of these Fusarium toxins on reproduction system and especially on spermatozoa after in vivo exposure. The aim of this study was to investigate the genotoxicity of FB1, DON and ZEA alone as well as in combination in low dosages on spermatozoa derived from breeding rabbit bucks after in vivo treatment. The rabbit bucks were fed for 65 days with the experimental diets; Control (C), FB1 (F), ZEA+DON (ZD) and FB1+ZEA+DON (FZD), the concentrations were 0 mg/kg, 5 mg/kg, 0.25+1 mg/kg and 5+0.25+1 mg/kg respectively. On day 65, semen was collected and Comet assay was performed to assess the genotoxicity (DNA damage). According to the Comet assay results, F treatment resulted in significantly less 0 comets compared to other treatments. Regarding score 1 all toxin treatments had similar proportions. As for score 2, F had significantly increased number of cells compared to FZD. Few cells had score of 3 (maximum 0.625%) whereas no cells with score 4 were found. It can be concluded that the combined toxins act rather antagonistically than FB1 alone since the combination led to lower DNA damage (higher score 0 and lower score 2). This is the first time interactive genotoxic effects of FB1, DON and ZEA on rabbit spermatozoa are reported.

Feed containing fumonisin (5 ppm; F), zearalenone (Z) and deoxynivalenol (D) (0.25 ppm+1 ppm; ZD) individually and these three toxins in combination (5 ppm+0.25 ppm+1 ppm; FZD) was fed to adult Pannon White (n =15/group) male rabbits (4±0.5 kg) for 65 days to determine the Fusarium toxin eff ect on breeding rabbit bucks’ sperm quality and endocrine function. Th e toxin levels were the lowest limit values for farm animals of the Commission Recommendation (2006/576/EC). On trial days 0, 30 and 65 blood and semen were sampled, and from semen pH, concentration, motility and morphology of the spermatozoa were investigated. Th e ratio of spermatozoa showing progressive forward motility decreased (P<0.05) from 80% to 67% in the FZD group. Diff erences were found between the data of the ZD (66.3%±23.7) and control animals (80.2%±11.2) concerning the normal morphology of spermatozoa. Aft er gonadotropin-releasing hormone analogue treatment, the testosterone concentration was lowered in the FZD animals aft er 65 days. Th ere was no diff erence among groups in feed consumption and BW. Histophatology revealed lowered spermiogenesis activity occurred in lower percentage in the ZD group (30.77%), while in FZD it was much more progressed (64.28%), referring to a synergistic eff ect of the three toxins.

T-2 toxin is the most acutely toxic trichothecene mycotoxin: it inhibits protein, DNA and RNA synthesis. The main goal of this study was to evaluate the rate of DNA damage caused by T-2 toxin in porcine mononuclear cells in increasing concentrations (0.1, 0.5 and 1.0 μmol) and after two different incubation periods (24 and 42 h). The lowest concentration caused DNA damage and about 50% of the treated cells could be categorised as having 1 to 4 scores in comet assay. In parallel with the increase of T-2 toxin concentration, the frequency of intact lymphocytes decreased from 50.2% (0.1 μM) to 36.3% (1.0 μM) in the first 24 h. In case of score 3, the highest concentration of T-2 toxin resulted in a 5-fold change, as compared to the lowest dose. Cells with score 4 were found only after exposure to 1.0 μM T-2 toxin. The exposure time did not have a significant effect on the results, while concentration did (P < 0.0001). However, a significant interaction between concentration and time as fixed factors (P < 0.0001) was found. When these were combined as a single factor, the results showed a significant toxin treatment effect on the results. It was concluded that a time- and dose-dependent DNA damaging effect of T-2 toxin could be demonstrated using peripheral blood mononuclear cells from healthy pigs by comet assay.

T-2 toxin is a secondary fungal metabolite produced by Fusarium species. Several in vitro and in vivo studies described genotoxic potential of T-2 toxin, which is generally accepted to be caused by oxidative stress. There are some data showing that colonic probiotic bacteria can remove mycotoxins via physical binding. Mannan oligosaccharides (MOS) are widely used animal feed to improve gastrointestinal health. Because of their interaction with microbes, the aim of our study was to determine the possible protective effect of MOS in T-2 toxicosis. Sucking rabbits were randomly assigned into two experimental groups, the control (C, n = 20) and the prebiotic (P, n = 20) group. In group P the feed of the does was completed with MOS. The young rabbits were allowed to consume the feed of the does from about the 17th days of age. The rabbits were weaned on the 35th day. At 7 weeks of age both groups (C and P) were divided into two (n = 10 in each), and half of the C and P rabbits were fed with the same diet as before, but contaminated with 2 mg/kg feed T-2 toxin (groups CT and PT). The animals were fed the toxin containing diet for the duration of 21 days. At the end of the 10th week blood samples were collected from 6 animals from group C, CT and PT. Mononuclear cells obtained from the rabbits were tested in comet assay to detect the genotoxic effect of T-2. All control samples (C) were negative in the test, i.e. all cells were scored as ‘0’. T-2 toxin in 2 mg/kg feed concentration had genotoxic effect on the rabbits’ lymphocytes, as could be concluded from the comet values. MOS supplementation in the feed had significant protective effect against T-2 as seen by the lower comet score frequencies compared to T-2 treated animals. These results demonstrate that MOS may reduce risk associated with the uptake of mycotoxins probably by their binding capacity or antioxidative properties.

The objective of this study was to evaluate the effect of supplementation of the growing rabbits’ diet by Spirulina or/and Thyme (food supplement) on the composition and amount of the microbiota and production of volatile fatty acids of the rabbits’ caecum. We used classical culturing methods and after bacterial DNA extraction, the quantity of bacteria (belonging to phylum Firmicutes and Bacteroidetes) were determined by qPCR reactions with the aid of bacterial ribosome coding DNA at the Molecular Biology Laboratory, University of Kaposvar. The experiment was carried out using rabbits (Pannon White) from rabbit-farm of Kaposvar University. Young rabbits were weaned on the 35th day after birth. All experimental animals were the same age. Duration of Spirulina and/or Thyme supplementation –after weaning- was 48 days. Samples were collected on the 14th, 28th and 48th day of the supplementation. No dietary effect on pH of the caecal content was detected. The number of Escherichia coli, total anaerobic and strictly anaerobic bacteria decreased by age, no effect of the diet could be demonstrated. Supplementation with Thyme resulted in slightly higher ratio of propionic acid, but the difference was not significant. In conclusion, Spirulina and/or Thyme supplementation of diet after weaning had no substantial effect on the volatile fatty acid (VFA) production, while the classical microbiological determination did not find a substantial effect on the composition of the caecal microbiota. By the Quantitative PCR method we measured significantly lower bacterial copy numbers in the samples of ST treated group. This indicates that the ST (Spirulina 5% and 3% Thyme) feed supplementation has an antimicrobial effect on the investigated bacterial groups in the caecum of growing rabbit.

Fusariotoxins are frequently existing contaminants in cereal and other plant products. High relationship was found between the levels of the trichothecene T-2 and HT-2 toxin detected in contaminated commodities. Many in vitro studies investigated the cytotoxicity of T-2 and HT-2, but the interaction between them has not been thoroughly studied yet. The aim of the study was to examine the dose dependent cytotoxic effect of T-2 and HT-2 toxins, single and in combination, using pig lymphocytes in the methyl thiazol tetrazolium (MTT ) assay. The mycotoxins were added at various concentrations, i.e. 0.5, 0.1, 0.05, 0.01 and 0.001 μM of T-2; 1.0, 0.5, 0.2., 0.1 and 0.05 μM of HT-2; and both mycotoxins combined. Two exposure times (6 and 24 h) were tested. Both T-2 and HT-2 toxins exerted a dose dependent effect. After 6 h incubation, the increase in concentration of T-2 from 0.001 to 0.5 μM and HT-2 for 0.05 to 1.0 μM resulted in lower cell viability by 22 and 17%, respectively. After 24 h cell viability was significantly lower compared to values obtained at 6 h, except 0.5 μM T-2 and 0.05, 1.0 μM HT-2, respectively. Measured cell viability for combinations of T-2 and HT-2 was higher compared to the calculated expected values. The two toxins caused lower cell survival when applied together than in single administration after 6 h incubation. After 24 h incubation this tendency was not consistent.

Summary Th e aim of the present study was to examine the chronic eff ect of T-2 toxin on feed consumption and sperm quality. Pannon White (n=10/group) male rabbits (weight: 4050-4500 g, age: 9 month) trained to ejaculate into artifi cial vagina were exposed to 0 (control), 0.05, 0.1 or 0.2 mg/animal/day of T-2 toxin by gavage for 63 days. On the 63rd day of the experiment semen was collected with an artifi cial vagina, and the following traits were evaluated: pH, concentration, morphology, motility with CASA, concentration of seminal plasma components such as citric acid, zinc and fructose. At the end of the experiment animals were necropsied and the testes were subjected to histopathological examination. T-2 toxin in 0.1 mg and 0.2 daily dose signifi cantly decreased feed intake in the fi rst two weeks but no signifi cant diff erence between groups were observed from the 4th week. Among the sperm quality traits examined only the ratio of spermatozoa with cytoplasmic droplets increased by 320% in animals treated with the highest dose of T-2. Th e 0.1 mg/animal/day toxin exposure resulted in a slight hyperplasia of the Leydig cells, while the highest dose (0.2 mg/animal/ day) caused hyperaemia, increased proliferative activity and hyperplasia of the Leydig cells. According to the preliminary results it seems, that adult male rabbits may tolerate the concentration of 0.05 mg/animal/day T-2 toxin.

The effects of intraperitoneally administered plant lectins were examined in rats and mice. Intraperitoneally injected ConA transiently decreased the leukocyte count in the peritoneal cavity, due to the agglutination and attachment of cells to the peritoneal lining. Subsequently the total cell count was increased for hours, exceeding initial values. Peritoneal fluid aspartate transaminase (AST) concentration showed little change during the accumulation of ascitic fluid. The most marked histological alterations were found when wheat germ lectin was injected ip. (WGA, 10 mg/kg, 6 h). Neutrophil granulocytes migrated across the wall of both arterioles and venules, but the response was highly variable among adjacent vessels. The wall of the arterioles may have impeded the migration of neutrophil granulocytes, resulting in their accumulation in the muscular layer. Granulocyte accumulation was also observed in patches under the mesothelium and in other sites of the interstitium. Marked dilatation and thrombosis of a few venules were also observed. Kidney bean lectin (PHA) induced similar but less pronounced changes. The neutrophil diapedesis suggests the release of mediator(s) from mesothelial cells and/or peritoneal white cells. The cytokine-induced neutrophil chemoattractant CINC-1, injected as control, resulted in the diapedesis of predominantly mononuclear cells in the omentum within 40 minutes. In rats ip. injected ConA increased the wet weight of spleen and liver within 6 and 10 h, respectively, but kidney weight did not change. Intravascular clumping of red blood cells, thrombosis and organ weight changes also suggest the absorption of ConA into the circulation. The experiments show that plant lectins, used as models of bacterial lectins, can reproduce some aspects of peritonitis.