The Molecular Toxicity Mechanism Of T-2 Toxin And Its Residues In Litopenaeus Vannamei

T-2 toxin produced by bacteria cell unit of sesquiterpene compounds,and widespread pollution in our country's a prawn feed,shrimp eat the feed contaminated with T-2 toxin,toxins will accumulate in prawn body cause the potential danger of aquaculture,residual toxins to enter the body through the food chain,food quality and safety problem.The prophase research for the prawn difference series of T-2 toxin harm,such as production performance,digestive system damage,abnormal detoxifying enzymes,restrain the immune system and antioxidant system and muscle quality.But about T-2 toxin less studies of biological macromolecules,biological macromolecules including lipid,protein,DNA and RNA,including prophase research has found that T-2toxins have bigger influence on the ?-3 unsaturated fatty acid;Group differences in learning the study of protein found T 2 toxin of arginine kinase difference big impact,but the differences between protein omics obvious shortcomings,easy cause large error,so you need to by Western blotting and RT-PCR experiments on the identification.And there is no research about DNA and RNA.About T-2 toxin in the prawn residual hazards research,the early stage of the group with T-2 toxin,hidden state T-2 toxin and toxin inside the prawn residue treatment RAW264.7 cells in mice,explore its RAW364.7JAK/STAT signaling pathways in cells key cytokine mRNA expression quantity.Only the quantity of mRNA expression is not fully prove that T-2 toxin,hidden state T-2 toxin and toxin in prawn residues of JAK/STAT signal pathway in the body the influence of key cytokines,need further by Western blotting experimental proof.In this study according to the principle of design of toxicology,prawns as the research object,setting different T-2 toxin exposure dose(0,1.2,2.4,4.8,12.2mg/kg.feed-1),making poison bait feeding prawn,detection of prawn DNA,the indicators of enzymes in the changes in the body.First of all,this study to investigate the DNA damage effect on T-2 toxin,the toxic effect with calf thymus DNA and T-2,T-2 toxin on viscosity of DNA and the influence of the absorbance at 260 nm,and through the response surface analysis to optimize the bestexperimental conditions;Prawn DNA extraction blank group,according to the optimum experiment conditions,the toxic effect of in vitro and T-2,T-2 toxin on viscosity of prawn DNA,and the influence of the absorbance at 260 nm.Detection of prawn DNA,extract infected dose group after 20 d accumulation then,different dose groups of infected shrimp body DNA viscosity and the change of absorbance at 260 nm.The results showed that T-2 toxin effect on viscosity of DNA is: the best conditions of T-2 toxin concentration 2.70 ng/mL;DNA concentration 50 mg/mL;The reaction time T for 43min;T-2 toxin effect on DNA OD260 values for: the best conditions of thick 0.70 ng/mL T-2 toxin;DNA concentration 60 mg/m L;The reaction time T for 30 min;T 2 toxin effect on viscosity of prawn DNA and OD260 value with calf thymus DNA has the same change;In the dose group infected shrimp viscosity of DNA in vivo and in vitro experiments have similarities,overall increase viscosity,but viscosity of DNA in 2.4 dose group was obviously lower blank group,and OD260 value change is not consistent.Secondly,on the early stage of the team by differences of proteomics found protein arginine kinase,by Western blotting detecting technology in different dosage group infected shrimp body arginine kinase expression,the quantity of protein and the difference point identified;Checked by RT-PCR technique dose group of prawn arginine kinase mRNA expression levels of change,further to its point of difference between protein identification.Found that Western blotting and rt-pcr experiments results,within the low dose group of arginine kinase expression quantity and amount of mRNA expression were lower,and higher in the high dose group,the experimental results and the difference is not in conformity with the results of proteomics.Finally,the extracted 12.2 dose group of infected shrimp hepatopancreas and muscle in the body hidden state T-2(m T-2s),free T-2 and T-2-GluA RAW264.7 cells for 24 h,respectively,by Western blotting technology to detect the JAK/STAT signaling pathways in cell signal factor(JAK1-3 and STAT1-3)the expression of quantity changes,and phosphorylation(p-JAK1-3 and p-STAT1-3).The results showed that: the free T-2 toxin and hidden state equal toxin promote cell signal factor expression,also can promote the phosphorylation of signal factor,the promoting effect is better than hidden state of free T-2.