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Abstract: Mycotoxins are a group of compounds produced by various fungi and excreted into the matrices on which they grow: often food intended for human consumption or animal feed. High toxicity and carcinogenicity of these compounds and their ability to cause various pathological conditions has led to widespread screening of foods and feeds potentially polluted with them. Maximum permissible levels in different matrices have also been established. As these are quite low, analytical methods for determination of mycotoxins have to be both sensitive and specific. In addition, an appropriate sample preparation and preconcentration method is needed to isolate analytes from rather complicated samples. In this article, an overview of analytical and sample preparation methods is given for the most often encountered mycotoxins in different samples, mainly in food. Special emphasis is on liquid chromatography with fluorescence and mass spectrometric detection, while in the field of sample preparation various solid-phase extraction approaches are discussed. However, an overview of other analytical and sample preparation methods less often used is also given. Finally, different matrices where mycotoxins have to be determined are discussed with the emphasis on their specific characteristics important for the analysis (human food and beverages, animal feed, biological samples, environmental samples). Various issues important for accurate qualitative and quantitative analyses are critically discussed: sampling and choice of representative sample, sample preparation and possible bias associated with it, specificity of the analytical method and critical evaluation of results.
 
Manuscript ID: IJMS-35-02
Type of Paper: Full Research Paper
Title: Pathogenesis and prevеntive measures in Mycotoxic nephropathies in farm animals and humans
Authors: Stoycho D. Stoev
Affiliations: Department of General and clinical pathology, Faculty of Veterinary Medicine, Trakia University, Students campus, 6000 Stara Zagora, Bulgaria, E-mail: stoev@uni-sz.bg
Abstract: Various aetiological factors contributing to the development of mycotoxic nephropathy in farm animals and humans are reviewed. The metabolism, elimination and mechanism of action of nephrotoxic mycotoxin ochratoxin A (OTA) are briefly described. The possible synergistic effects between OTA and other mycotoxins are also considered. The most convenient ways of prophylaxis and various preventive measures are reviewed. A reference was made concerning the most successful methods of veterinary hygiene control in the slaughterhouses in order to prevent the entering of OTA in commercial channels with a view to human health. The economic efficacy of these prophylactic procedures is also considered
  
Manuscript ID: IJMS-35-04
Type of Paper: Review
Title: Health hazards and Management of mycotoxicosis: A critical review
Authors: Hussein A Sheashaa, Khaled M Mahmoud and Mohamed A Sobh
Affiliations: Nephrology Unit, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt
Abstract: Background. Mycotoxins are toxic metabolites produced by fungi, Serveral mycotoxins in agricultural products cause health hazards to people and animals and lead to many economical problems. Dangerous mycotoxins are naturally present in foods and in the environment. They are pathologically classified as hepatotoxins, nephrotoxins, vomitoxin and neuro-musculotoxin, some of which are potentially carcinogenic and mutagenic. Prevention and Treatment. Several effective ways for prevention and control of fungi and their dangerous mycotoxins have been presented. The methods include biological control and physical and chemical treatments. Selection of fungal resistant hybrids of crops are recommended and further experimented. Pre-harvesting preparation of the field and environments should be aware of. Drying of commodities after post harvest is the most economical and effective means for farmers or layment, but sometimes is not suitable during rainy season or wet condition. Thermal treatment or gamma irradiation is not effective or practically used by villagers. Chemical treatments such as alkalinization and ammoniation are well-recognized and industrially used. Some modifications of the application of effective chemicals to the detoxification of mycotoxins should be developed. International cooperations through authorized organizations should be promoted and supported aiming the benefits for the economics and health of people of all the nations.
  
Manuscript ID: IJMS-35-05
Type of Paper: Full Research Paper
Title: Fumonisins and Trichothecenes in Cereals, Invisited
Authors: Selma Yazar and Gülden Omurtag
Affiliations: Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Marmara University, 34668, Haydarpasa - Istanbul, Turkey
Abstract: Fumonisins are phytotoxic mycotoxins which are synthesized by various species of the fungal genus Fusarium species such as Fusarium verticillioides (Sacc.) Nirenberg (ex F. moniliforme Sheldon) and Fusarium proliferatum. The trichothecene (TC) mycotoxins are secondary metabolites produce by several fungal genera, especially Fusarium, Stachybotrys, Trichoderma, Memnoniella and Myrothecium. Fusarium mycotoxins are widely dispersed in cereals and their products. Fumonisins and the trichothecenes (TCs) are hazardous for human and animal health. Contamination with TCs causes a number of illnesses in human and animals such as decrease in food consumption (anorexia), depression or inhibition on immun system function and haematoxicity. The purpose of this paper is to give a review of the papers published on the field of fumonisin and trichothecene mycotoxins in cereals consumed in the world.
 
Manuscript ID: IJMS-35-06
Type of Paper: Full Research Paper
Title: Resveratrol Prevents Citrinin-Induced Apoptotic Biochemical Signaling in Human Hepatoma G2 Cells
Authors: Ching-Yu Lai and Wen-Hsiung Chan
Affiliations: Department of Bioscience Technology and Center for Nanotechnology, Chung Yuan Christian University, Chung Li, Taiwan 32023
Abstract: The mycotoxin citrinin (CTN), a natural contaminant in foodstuffs and animal feeds, has cytotoxic and genotoxic effects on various mammalian cells. CTN is known to cause cell injury, including apoptosis, but the precise regulatory mechanisms of CTN action are currently unclear. Resveratrol, a member of the phytoalexin family found in grapes and other dietary plants, is known to have antioxidant and anti-tumor properties. In the present study, we examined the effect of resveratrol on apoptotic biochemical events caused by incubation of Hep G2 cells with CTN. Resveratrol inhibited the CTN-induced ROS generation, activation of JNK, loss of mitochondrial membrane potential (MMP), activation of caspase-9, caspase-3 and PAK2, and apoptosis. Study also found that both resveratrol and ROS scavengers (i.e., NAC and α-tocopherol) abolished CTN-stimulated intracellular oxidative stress and apoptosis. In addition, experiments further found that activation of JNK is required for CTN-induced mitochondria-dependent apoptotic biochemical changes such as loss of MMP, activation of caspases and PAK2. Moreover, we also found that PAK2 activity is required for CTN-induced apoptosis. These results demonstrated that CTN triggers apoptosis in Hep G2 cells via ROS generation and JNK activation for activation of mitochondria-dependent cell death signaling and these apoptotic biochemical events were blocked by pretreatment with resveratrol through its antioxidant property.
 
Manuscript ID: IJMS-35-07
Type of Paper: Full Research Paper
Title: Influence of the trichothecenes DON and T-2 toxin on physico-chemical characteristics in a fermented product with Saccharomyces cerevisiae
Authors: Jaqueline Garda-Buffon and Eliana Badiale-Furlong
Affiliations: Mycotoxins Laboratory, Chemistry Department, Federal University Foundation of Rio Grande (FURG), Rio Grande do Sul – Brazil,
Abstract: The aim of this work was to evaluate the influence of these toxins during the fermentation process on the characteristics of a fermented product with Saccharomyces cerevisiae. Alcoholic fermentation on a laboratory scale was carried out using malt contaminated with DON and T-2 toxin in a range of 0 to 1000ng/g. The process lasted 120 hours and was carried out at 14ºC. The levels of DON and T-2 toxin in the fermented liquid phase were around 41% of the initial contamination. T-2 toxin had some significant effects, increasing the consumption of reducing sugars and leading to a decrease in pH when compared to the values observed when it was absent. The alcohol profile was altered in the presence of the studied mycotoxins, being this variation best explained by T-2 toxin.

Type of Paper: Full Research Paper
Title: Production and Comparison of Monoclonal Antibodies against Fumonisin B₁ and Recombinant Single Chain Variable Fragment
Authors: Young-Jin Cho †, Jun-Bock Park †, Won-Ki Min, Yong-Cheol Park and Jin-Ho Seo *
Affiliations: Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Republic of Korea; † These authors equally contributed to the work. * Corresponding author: E-mail: jhseo94@snu.ac.kr; Tel: +82-31-290-2583; Fax: +82-31-293-4789
Abstract: A monoclonal antibody (mAb) against fumonisin B1 (FMB1) was generated from the hybridoma cell line D8A54, which was established by the fusion of Sp2/0-Ag14 myeloma cells with spleen cells isolated from a BALB/c mouse immunized with a FMB1 conjugated to bovine serum albumin (FMB1-BSA). An isotype of this mAb was identified as IgG1 class in the heavy chain and kappa class in the light chain. The detection limit of 14nM was determined and 50 % binding inhibition was reached at 305 nM free FMB1 by a competitive direct enzyme-linked immunosorbent assay (cdELISA). No cross-reactivities to other mycotoxins, such as ochratoxin A (OTA), aflatoxin B1 (AFB1) and deoxynivalenol (DON) were observed, suggesting a highly specific FMB1 antibody. A recombinant single chain variable fragment antibody (scFv) has been cloned from the hybridoma cell and was expressed as inclusion bodies in recombinant Escherichia coli (E. coli). The insoluble scFv antibody against FMB1 was refolded and used to determine the FMB1 binding activity. The refolded scFv had about 10-fold lower binding affinity compared with the FMB1-specific  monoclonal antibody.
 
Type of Paper: Full Research Paper
Title: Isolation and Characterization of Ochratoxin A and Aflatoxin B1 producing Fungi infecting Grapevines cultivated in Tunisia
Authors: S. Melki Ben Fredj 1,*, F. Mathieu2 , A. Lebrihi2, S. Chebil1, A. Mliki1
Affiliations: 1 Technopark borj-Cédria (ex INRST). Centre de Biotechnologie, Laboratoire de Physiologie Moléculaire de la Vigne (LPMV), B.P. 901, Hammam-Lif 2050. Tunisie; 2 Ecole Nationale Supérieure Agronomique de Toulouse (ENSAT), Laboratoire de Génie Chimique UMR 5503 (CNRS/INPT/UPS), A. de l’Agrobiopôle BP 107, Auzeville-Tolosane 31450- France.
Abstract: Toxigenic potential of OTA and AFB1 have been evaluated for fungal strains harvested from different vineyards in Tunisia. Among 100 strains of Aspergillus isolated and identified. The most frequent strains were the Aspergillus niger aggregate (72%). 5% were confirmed as OTA producers. The high OTA potential producers were Aspergillus carbonarius (80% of them). Among 100 Penicillium isolates, we revealed P. chrysogenum (58%), P. brevicompactum (13%), P. verruculosum (22%), P. glabrum (5%), P. minioluteum (2%). Only 89 % of P. chrysogenum and 77% of P. verruculosum were able to produce OTA. 23% of Aspergillus flavus were identified with significant amounts of aflatoxin B1 production (39%). The quantification of this mycotoxin (AFB1) shows concentrations between 0.021 and 0.054 mg/g. The study shows that Tunisian grapes can be contaminated by OTA and AFB1.

Type of Paper: Full Research Paper
Title: Synergistic and/or antagonistic actions of Beauvericin and T-2 mycotoxins on antioxidant systems in tomato seedlings
Authors: Costantino Paciolla and Antonella Florio
Affiliations: Department of Plant Biology and Pathology, University of Bari, Italy
Abstract: During their life cycle plants can undergo simultaneous attack by different pathogens. It is well known that in various plant-pathogen interactions mycotoxins are produced which have an important role in pathogenesis. In addition it has been shown that many phytotoxins are responsible for ROS production. Plants counteract the overaccumulation of ROS by reinforcing defence systems such as the ascorbate-glutathione and other antioxidant systems. In our study we have used T-2 and Beauvericin, two mycotoxins with different action mechanisms produced by several Fusarium species. Since the comprehension of the mechanisms of mycotoxin actions on host plants at the cellular and biochemical level is very important to prevent the development of the diseases, we have studied in tomato plants both single and combined effects of the two mycotoxins on some defence systems. So, we analyse the ascorbate-glutathione cycle components and the POD enzymes. Moreover, we determine the H2O2 content, as an index of oxidative stress. The possible synergistic and/or antagonistic actions of Beauvericin and T-2 were discussed.

Type of Paper: Review
Title: Effects of milling and cooking process on the deoxynivalenol content in wheat
Authors: Masayo Kushiro, Hiroyuki Nakagawa, Hitoshi Nagasima
Affiliations: National Food Research Institute, 2-1-12 Kannondai, Tsukuba 305-8642, Japan
Abstract: Deoxynivalenol (DON) is a natural-occuring mycotoxin mainly produced by Fusarium graminearum, a food-borne fungi widely distributed in crops and occur as one of the most important mycotoxins in wheat and wheat-based foods and feeds. DON belongs to a group of trichothecene mycotoxin. DON and other trichothecenes affect animal and human health causing diarrhea, vomiting, gastro-intestinal inflammation and immunomodulation. The toxic mode of action of DON is considered to be a noncompetitive inhibition of the biosynthesis of proteins in eukaryotic cells. Since the rate of the occurrence of DON in wheat is high, effective procedures to remove or eliminate DON from food products is essential to minimize exposures in those who live on wheat. This paper reviews recent researches on the effects of milling and cooking on the concentration of DON as well as the changes on the toxicity.

Type of Paper: Review
Title: Mycotoxin detection as cops and robbers play: is it suitable to play?
Authors: Chiara Dall’Asta, Gianluigi Ingletto and Pietro Cozzini
Abstract: As in a cops and robbers play we discover new mycotoxins, new food pollutants, new xenoestrogens everyday and we are forced to develop quicly new molecules as chemo- or biosensors or to modify existing  molecules able to recognize these new pollutants. Sometime new techniques and new protocols are required. For this reason, there is a real need for rapid, sensitive and inexpensive sensors for the detection of toxic pollutants along the food processing chain to obtain real-time monitoring data on contamination, which may be use for  food safety assessment. This will result in an enormous cost saving to agro-food industry through the prevention and reduction of product recalls and reduced treatment costs.
Many technologies for detecting low molecular weight pollutants exist. Those that have recently been applied to xenobiotics detection include: evanescent wave technologies, lateral flow and dipstick devices, fluorescence polarization, microbead assays, flow injection liposome immunoassays, capillary electrophoretic immunoassays, and flow injection lipid bilayer assays. All these techniques require an immunochemical approach based on an antigen-antibody interaction. Although their high specificity against the analyte, these devices show several drawbacks. Immunoassay such as dipsticks or ELISA kits should be stored and used under controlled environmental conditions and may give false positive/negative results due to matrix effects.
Thus, there is a need of “smart molecules” showing biomimetic properties towards food pollutants and xenobiotics. These compounds could exhibit a substrate-selective recognition mechanism similar to that of antibodies or enzymes, being at the same time more stable and affordable. The development of such biomimetic compounds requires a strong synthetic effort which should be driven by a sound molecular design to allow a target-tailored approach. Molecular models are mostly used for two reasons, the comprehension of some experimental data in lack of structural informations or the design and the optimization of new compounds to drive new synthesis. Here we present a brief review of the methods used to detect mycotoxins and food pollutants, considering goodness and limits. Then we proposed a new methodology, fast, efficient and cheap, based on a combination of computer chemistry aided design and fast fluorescence, that can help to drive synthesis in a more efficient way.

Title: Trichothecenes: transformation reactions and their applications in detoxification
Authors: Jianwei He1, Ting Zhou ^1,*,  J. Christopher Young ¹ and Greg J. Boland ²
Affiliations: (1) Guelph Food Research Centre, Agriculture and Agri-Food Canada, Guelph, ON, N1G 5C9, Canada  zhout@agr.gc.ca; (2) Department of Environmental Biology, University of Guelph, Guelph, ON, N1G 2W1, Canada
Abstract: Trichothecenes are a group of structure related mycotoxins that commonly contaminate cereals worldwide.  Trichothecenes can cause acute and chronic toxicity to a wide range of organisms, including human beings and livestock.  They bind to eukaryotic ribosomes, block translation and inhibit the elongation of peptide chains, and thus, inhibit protein synthesis.  Physical decontamination techniques included cleaning, sorting, separation, thermal inactivation, irradiation, ultrasound, and adsorption have been developed to reduce the concentration of trichothecenes in affected grains.  However, these methods cannot degrade or completely remove the toxins, and usually cause losses of nutrients.  Approaches based on structural modification of trichothecenes to less or non-toxic chemicals may provide an improved means to treating trichothecenes-contaminated grains because they potentially can overcome some of these limitations and be more efficient.  This paper describes the transformation reactions and toxicity of transformation products of trichothecenes in chemical and biological systems.  Efforts will focus on discussing the current and potential applications in detoxification of trichothecenes using chemicals, enzymes, microorganisms and organisms related to these reactions.