Preliminary Study On Toxicity Reduction Method Of Tetrodotoxin

China's puffer fish farming industry was originally launched in 1990.After nearly30 years of development,China has made remarkable achievements in puffer fish farming.However,puffer fish contains a deadly neurotoxin called tetrodotoxin(TTX),in which the TTX content of the puffer fish liver and ovary is relatively high.TTX is a highly toxic neurotoxin that is a typical Na~+channel inhibitor,and 0.5 mg can kill people.Since the Ministry of Agriculture's General Office(Nongjian Fishing[2016]No.53)has conditionally liberalized the processing and management of farmed puffer fish,the market for puffer fish products has gradually increased.However,according to the regulations,only the puffer muscle tissue processed by qualified production enterprises or restaurants can be sold as edible raw materials and products,and the visceral tissues are not edible.The puffer fish liver is rich in unsaturated fatty acids and eight essential amino acids and other nutrients.Therefore,it is necessary to carry out research on resource recycling of puffer liver tissue.In this study,we aimed to reduce the toxicity of TTX in puffer fish viscera with the aim of recycling and reusing puffer fish viscera,in order to reduce the toxicity of TTX in puffer fish visceral tissues and the development of unsaturated fatty acids such as EPA,DHA and oleic acid.Use to provide scientific evidence.At present,there are few reports on the toxicity reduction of TTX at home and abroad.The research shows that the viscera viscera can reach the non-toxic level after treatment with NaOH for 15 min,but the treatment method has a great damage to the nutritional quality of fatty acids and proteins in the viscera.Japanese nationals discovered that after two years of pickling of puffer fish porpoise,the ovaries were non-toxic and edible.In view of this research background,this study used alkaline chemicals and microbial methods to study the toxicity reduction method of TTX in puffer liver tissue.Firstly,the alkaline chemicals and strains with the ability to reduce TTX toxicity were screened,and then the consumption of baking soda and Lactobacillus rhamnosus with better depletion effect was optimized to optimize the TTX toxicity reduction condition by competitive ELISA and magnetic immunochromatographic test.Paper strips and mouse bioassay were used to analyze the changes in the content and toxicity of TTX before and after subtraction.The content of unsaturated fatty acids in the liver of puffer fish after fermentation was analyzed by gas chromatography-mass spectrometer.The toxicity reduction method of TTX was preliminarily discussed.The main findings of this paper are as follows:1.Screening substances that can reduce TTX toxicity from commercially available alkaline chemicals,using competitive ELISA and classical mouse biological methods to analyze changes in TTX content and toxicity,and analyzing the porpoise tissue after toxicity reduction by gas chromatography-mass spectrometer Changes in the content of unsaturated fatty acids(EPA,DHA and oleic acid).Competitive ELISA results showed that when 10.0%(w/v)of edible baking soda was added,the TTX content was reduced by 59.64%after soaking for 72 h at 25?.Mouse bioassay results show that consumption of baking soda has the ability to reduce TTX toxicity.The results of gas chromatography-mass spectrometer showed that the total amount of DHA and EPA and oleic acid in the tissues of wild puffer fish treated with baking soda for 72 h increased by 6.67%and 28.27%,respectively,indicating that the consumption of baking soda can not destroy the puffer fish.Under the premise of unsaturated fatty acids in tissues,the toxicity in the liver is reduced.2.The common probiotics and their products were selected to reduce the toxicity of puffer fish tissue.The magnetic immunochromatographic test strips and mouse biological methods were used to analyze the TTX content and toxicity changes before and after subtraction.The qualitative results of magnetic immunochromatographic test strips showed that Lactobacillus rhamnosus,Bifidobacterium and Lactobacillus could reduce the toxicity of TTX.The quantitative results of test strips showed that the Lactobacillus rhamnosus had a toxicity reduction rate of 92.46%for puffer fish tissue.The reduction effect is the best.The results of the mouse bioassay showed that Lactobacillus rhamnosus reduced TTX toxicity to a non-toxic level.It indicated that the reduction of TTX toxicity by Lactobacillus rhamnosus was stronger than that of baking soda.3.To study the TTX toxicity of Lactobacillus rhamnosus in the liver of wild puffer fish,and to analyze the content and toxicity of TTX before and after the use of competitive ELISA,magnetic immunochromatographic test strip and mouse biological method.The change of unsaturated fatty acid content in the liver of puffer fish at different fermentation time was analyzed by gas chromatography mass spectrometer serial instrument,in order to establish a toxicity reduction method of TTX.The competitive ELISA results showed that when the amount of Lactobacillus rhamnosus was 8%(v/v)and the shaking speed was 180 rpm/min,the content of TTX was decreased by 92%after 1 h of fermentation.The mouse bioassay showed that the wild puffer fish liver after 7 days of fermentation by Lactobacillus rhamnosus could be reduced to a non-toxic level.In addition,gas chromatography-mass spectrometry showed that the EPA and DHA in the liver decreased by 11.93 and 22.5%,respectively,and the oleic acid content increased by 34.66%.The above results indicate that Lactobacillus rhamnosus can reduce the toxicity of TTX to a non-toxic level,although this treatment will reduce the content of EPA and DHA,but it is within an acceptable range.In summary,this study found that both alkaline chemicals and probiotics can reduce the toxicity of TTX.The toxicity reduction effect of baking soda on TTX is lower than that of Lactobacillus rhamnosus,but this treatment helps the dissolution of EPA,DHA and oleic acid in the liver;the fermentation treatment of puffer liver by Lactobacillus rhamnosus can make it The toxicity is reduced to a non-toxic level.Although the treatment will decrease the content of EPA and DHA,the oleic acid content in the tissue increases.Considering the content of unsaturated fatty acid,the content of unsaturated fatty acid is basically unchanged and can be used as a feed additive for regeneration.Resources have good market potential and application value.