Evaluation of the Antioxidant, Anti-Inflammatory and Cytoprotective Activities of Halophyte Extracts against Mycotoxin Intoxication

Twelve halophyte species belonging to different families, widely represented along French Atlantic shoreline and commonly used in traditional medicine, were screened for protective activities against mycotoxins, in order to set out new promising sources of natural ingredients for feed applications. Selected halophytic species from diverse natural habitats were examined for their in vitro anti-mycotoxin activities, through viability evaluation of Madin-Darby Bovine Kidney (MDBK) and intestinal porcine enterocyte (IPEC-J2) cell lines. Besides, the in vitro antioxidant activities of plant extracts were assessed (total antioxidant and 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging bioassays). Of the 12 species, Galium arenarium, Convolvulus soldanella and Eryngium campestre exhibited the most protective action on MDBK and IPEC-J2 cells against zearalenone (ZEN) or T2 toxin contamination (restoring about 75% of cell viability at 10 μg·mL−1) without inflammation response. They also had strong antioxidant capacities (Inhibitory concentration of 50% (IC50) < 100 μg·mL−1 for DPPH radical and total antioxidant capacity (TAC) of 100 to 200 mg Ascorbic Acid Equivalent (AAE)·g−1 Dry Weight), suggesting that cell protection against intoxication involves antioxidant action. A bio-guided study showed that fractions of G. arenarium extract protect MDBK cells against T2 or ZEN toxicity and several major compounds like chlorogenic acid and asperuloside could be involved in this protective effect. Overall, our results show that the halophytes G. arenarium, C. soldanella and E. campestre should be considered further as new sources of ingredients for livestock feed with protective action against mycotoxin intoxication.

Immunochromatographic Tests for Mycotoxins Detection with the Use of Ultrasmall Magnetite Nanoparticles

The use of small (with a diameter of 7–12 nm) superparamagnetic Fe3O4 nanoparticles as carriers for antibodies in lateral flow immunoassay is considered. Increased total surface area for such suspension provides a concentration of analytes with an increase in their concentration up to 50 times. When the concentrated complexes were redissolved, aggregates with diameters of 100–500 nm were obtained, serving as colored markers in the assay. Further, magnetite–antibodies complexes are more tolerant to methanol (up to 30%) than native antibodies, thus providing minimal dilution of tested extracts. Lateral flow tests for mycotoxins zearalenone, T2-toxin, and aflatoxin B1 were developed and demonstrated applicability to control food products and raw materials.

Investigating Multi-Mycotoxin Exposure in Occupational Settings: A Biomonitoring and Airborne Measurement Approach

Investigating workplace exposure to mycotoxins is of the utmost importance in supporting the implementation of preventive measures for workers. The aim of this study was to provide tools for measuring mycotoxins in urine and airborne samples. A multi-class mycotoxin method was developed in urine for the determination of aflatoxin B1, aflatoxin M1, ochratoxin A, ochratoxin α, deoxynivalenol, zearalenone, α-zearalenol, β-zearalenol, fumonisin B1, HT2-toxin and T2-toxin. Analysis was based on liquid chromatography–high resolution mass spectrometry. Sample pre-treatments included enzymatic digestion and an online or offline sample clean-up step. The method was validated according to the European Medicines Agency guidance procedures. In order to estimate external exposure, air samples collected with a CIP 10 (Capteur Individuel de Particules 10) personal dust sampler were analyzed for the quantification of up to ten mycotoxins, including aflatoxins, ochratoxin A, deoxynivalenol, zearalenone, fumonisin B1 and HT-2 toxin and T-2 toxin. The method was validated according to standards for workplace exposure to chemical and biological agents EN 482. Both methods, biomonitoring and airborne mycotoxin measurement, showed good analytical performances. They were successfully applied in a small pilot study to assess mycotoxin contamination in workers during cleaning of a grain elevator. We demonstrated that this approach was suitable for investigating occupational exposure to mycotoxins.

Mycotoxins in Maize and Implications on Food Security: A Review

Mycotoxin poisoning is not restricted to pets and farm animals, it causes diseases and death in humans. Mycotoxin producing fungi are common components of the epiphytic and endophytic microflora in crops resulting in natural crop contamination in the field and during storage. The level of contamination is influenced by the genetics of the plant and fungi, management practices and prevailing climatic conditions. The Global movement of maize products necessitates global as well as country-specific surveys on mycotoxin occurrence. Significant differences in the concentrations of deoxynivalenol, fumonisins and zearalenone were found in commercial maize for the seven years under review but no significant difference was detected between white and yellow maize types with regards to fumonisins and zearalenone concentrations. The absence of Aflatoxins, Ochratoxin-A, T2-toxin and HT-2 toxin in the commercial maize samples from 2010/2011 to 2016/2017 seasons is a food safety advantage for South Africa maize producers.

The T2 Toxin Produced by Fusarium spp. Impacts Porcine Duodenal Nitric Oxide Synthase (nNOS)-Positive Nervous Structures—The Preliminary Study

T2 toxin synthetized by Fusarium spp. negatively affects various internal organs and systems, including the digestive tract and the immune, endocrine, and nervous systems. However, knowledge about the effects of T2 on the enteric nervous system (ENS) is still incomplete. Therefore, during the present experiment, the influence of T2 toxin with a dose of 12 µg/kg body weight (b.w.)/per day on the number of enteric nervous structures immunoreactive to neuronal isoform nitric oxide synthase (nNOS—used here as a marker of nitrergic neurons) in the porcine duodenum was studied using the double immunofluorescence method. Under physiological conditions, nNOS-positive neurons amounted to 38.28 ± 1.147%, 38.39 ± 1.244%, and 35.34 ± 1.151 of all enteric neurons in the myenteric (MP), outer submucous (OSP), and inner submucous (ISP) plexuses, respectively. After administration of T2 toxin, an increase in the number of these neurons was observed in all types of the enteric plexuses and nNOS-positive cells reached 46.20 ± 1.453% in the MP, 45.39 ± 0.488% in the OSP, and 44.07 ± 0.308% in the ISP. However, in the present study, the influence of T2 toxin on the intramucosal and intramuscular nNOS-positive nerves was not observed. The results obtained in the present study indicate that even low doses of T2 toxin are not neutral for living organisms because they may change the neurochemical characterization of the enteric neurons.

The Effect of Probiotics on T-2 Mycotoxin Induced Apoptosis in Chicken Liver Tissue

In recent years many researchers have described the reduced mycotoxin toxicity caused by probiotic bacteria. Since reduction under gastrointestinal conditions of the bioavailability of mycotoxins by probiotics is not fully investigated in birds, the aim of the study was to determine the effect of probiotic on T-2 mycotoxicosis induced apoptosis in broiler’s liver. For the study, twelve 1-days old broilers were divided equally into T-2 toxin (T2) and probiotic with T-2 (P+T2) groups. From the first experimental day, probiotic Enterococcus faecium DSM 7134 was administered in drinking water to P+T2 group. From the fourth day, T-2 toxin was given for three consecutive days to T2 toxin group. At 8th experimental day chicken were sacrificed, liver was fixed in buffered 10% formalin, embedded into paraffin, slices 5 μm in thickness were cut followed by immunohistochemical staining with polyclonal primary antibodies p21 and p53 (Abcam, UK) according to the manufacturers’ guidelines (IHC kit, Abcam, UK). Blood samples were taken by cardiac puncture to measure liver enzymes. Immunohistochemical staining revealed strong expression of p53 and p21 antibodies in hepatocytes nuclei as well as around blood vessels in T-2 toxin group’s chicken liver tissue. Staining by both antibodies was less intensive in P+T2 group. Enzyme analysis showed significantly increased (p<0.05) blood aspartate transaminase and alanine transaminase concentrations by 33.87% and 68.03% respectfully in T2 toxin group, while enzyme concentrations were decreased in P+T2 group. The obtained results showed reduced features of liver apoptosis in treatment with probiotic bacteria.

Assessment of the quality and safety of carpets with mycotoxicosis

Introduction. The article deals with the results of the mycotoxicological analysis of feed samples of pond fish. There were found the T2-toxin and aflatoxins.The combined presence of aflatoxins B1, B2, G1 and G2 and T2-toxin was found in all samples of fish feed. Was also studied the impact of combination of several mycotoxins on organoleptic and physico-chemical characteristics of fish carcasses. Fish, affected by mycotoxins, can be classified to the category of doubtful freshness. Materials and methods of research. The studies were conducted between May and October 2019. The investigated materials were a large number of grain and grain mixtures, for feeding fish in six ponds in Nikolaevska settlement community of Sumy region. Preparation of grain samples were made according resolution of the Cabinet of Ministers of Ukraine, June 14, 2002, No. 833 «The order of the selection of samples animal, plant and biotechnological origin». The studies of the toxicity and persistent T2-toxin and full amount of aflatoxins B1, B2, G1 and G2 are conducted on the basis of the Sumy Regional Laboratory of Veterinary Medicine. Feed toxicity was investigated by bioassay on the Tetrahymena piriformis infusorium used the DSTU 3570-97. The RIDASCREEN test systems were used to investigate the aflatoxins B1, B2, G1 and G2 in grains and grain mixtures. Ichthyopathological studies were conducted in the department of Veterinary and Sanitary Examination, Microbiology, Zooghygiene, Safety and the Guality of Livestock Products of the Sumy National Agrarian University by the accepted methods. Results of research and discussion. In the territory of the Nikolaevska settlement community of Sumy region there are six ponds, in which hold fish, mainly crucian and carp. The fish are fed once a week. Feed mainly consists of grain waste (barley, wheat), which is supplied by local farmers. Food is stored in unsuitable premises. In the feed we noted the presence of remains of substandard grain, which was thrown out during separation. Grain samples are not sampled and laboratory tests are not carried out. The feed is shipped by transport to ponds, dumped from the shore into a pond, immersed in to the water, where consumed by the fish. Totaly 19 samples of feed were examined during this period. Organoleptic evaluation of grain and grain mixtures revealed that the color of the grain was natural, the smell of all the samples had a certain tinge of moldy-musty, some samples had signs of fermentation and mold. Aqueous solutions of the grain extracts and mixtures of the test samples caused a stopping of movement and death of all Tetrahymena piriformis infusions up to 60 minutes in 7 samples, that indicates the toxicity of these feeds. Low toxicity was detected in 11 samples of experimental feed. The absence of toxicity was evidenced by the activity of Tetrahymena piriformis infusoriums, which persisted for 1 hour after the action of aqueous extracts of the samples. The results of the determination of the toxicity of grain feed using infusorium Tetrahymena piriformis. To analyze the content of major mycotoxins was used competitive enzyme immunoassay. The RIDASCREEN FAST Aflatoxin test system and the RIDASCREEN FAST T-2 Toxin test system have a high sensitivity of 0.0017 mg / kg and 0.05 mg / kg, respectively, which made it possible to determine the content of T-2 toxin, the amounts of aflatoxins B1, B2, G1 and G2 in their lowest concentration. The results of analysis of feed samples using the amount of aflatoxins B1, B2, G1 and G2 That in all samples of grain and grain mixtures was established the presence of aflatoxins B1, B2, G1 and G2. In 7 samples its content exceeds the maximum permissible concentrations. In 10 samples, the combined presence of aflatoxins B1, B2, G1 and G2 and T2-toxin was found, in 5 samples - exceeding the maximum levels. It is known that the combination of several mycotoxins can lead to their synergistic interaction, which will have a more pronounced toxic effect. Unfortunately, it was not possible to determine the maximum content of aflatoxins B1, B2, G1 and G2 and T2-toxin, so further test kits with different reading ranges should be used, since highly sensitive mycotoxin detection kits make it impossible to determine up to 1 mg / kg. In the following research, we studied the chemical properties of fish meat: reaction to peroxidase (benzidine sample), amount of amino-ammonia nitrogen, reaction with copper sulfate, reaction with Eber reagent, determination of hydrogen sulfide, pH and reaction with Nesler reagent. When muscle is damaged by mycotoxin, the products protein breakdown appears, which promotes rapid breakdown of tissue elements and leads to rapid deterioration of fish. Analyzing changes in veterinary-sanitary and physico-chemical parameters of fish affected by mycotoxicosis, we can classify the affected fish in the category of doubtful freshness. Conclusions and prospects for further research. During the investigate of grain and grain mix for feeding pond fish it was found that 19 samples were highly toxic and 36.8% were law toxoc. 2. Using the competitive enzyme immunoassay the combined presence of aflatoxins B1, B2, G1 and G2 and T2-toxin was found in all samples In 7 samples its content exceeds the maximum permissible concentrations. T2-toxin was found was found in 16 samples, and in 10 samples its content exceeds the maximum permissible concentrations. Fish affected by mycotoxins for organoleptic and physico-chemical characteristics can be classified to the category of doubtful freshness.

Plant biotransformation of T2 and HT2 toxin in cultured organs of Triticum durum Desf

The present study aimed at elucidating the uptake and biotransformation of T2 and HT2 toxins in five cultivars of durum wheat, by means of cultured plant organs. An almost complete absorption of T2 toxin (up to 100 µg) was noticed after 7 days, along with the contemporaneous formation of HT2 in planta, whereas HT2 showed a slower uptake by uninfected plant organs. Untargeted MS-analysis allowed to identify a large spectrum of phase I and phase II metabolites, resulting in 26 T2 and 23 HT2 metabolites plus tentative isomers. A novel masked mycotoxin, 3-acetyl-HT2-glucoside, was reported for the first time in wheat. The in vitro approach confirmed its potential to both investigate the contribution of plant metabolism in the biosynthesis of masked mycotoxins and to foresee the development of biocatalytic tools to develop nature-like mixtures to be used as reference materials.

Use of the Secreted Proteome of Trametes versicolor for Controlling the Cereal Pathogen Fusarium langsethiae

Fusarium langsethiae is amongst the most recently discovered pathogens of small grains cereals. F. langsethiae is the main producer, in Europe, of T2 and HT-toxins in small grain cereals, albeit often asymptomatic; this makes its control challenging. The European Union (EU) is pushing hard on the use of biocontrol agents to minimize the use of fungicides and pesticides, which are detrimental to the environment and responsible for serious pollution of the soil and superficial water. In line with EU directives (e.g., 128/2009), here we report the use of protein fractions, purified from the culture filtrate of the basidiomycete Trametes versicolor, for controlling F. langsethiae. T. versicolor, a so-called medicinal mushroom which is applied as a co-adjuvant in oncology and other pathologies as a producer of biological response modifiers. In this study, the exo-proteome of T. versicolor proved highly efficient in inhibiting the growth of F. langsethiae and the biosynthesis of the T2 toxin. Results are promising for its future use as a sustainable product to control F. langsethiae infection in cereals under field conditions.