【摘要】：T-2毒素被認(rèn)為是A族單端孢霉烯族毒素中毒性最強(qiáng)的,由于其高毒性特性,曾被用于戰(zhàn)爭(zhēng)作為生化武器。某些真菌感染谷物后在適宜的條件下很容易產(chǎn)生真菌毒素。由于植物性成分在水產(chǎn)飼料中所占的比例不斷增加,T-2毒素對(duì)水產(chǎn)動(dòng)物的危害也在不斷增加,這不但能造成水產(chǎn)品質(zhì)量品質(zhì)的下降,同時(shí)能帶來(lái)嚴(yán)重的食品安全問題。因此,研究水產(chǎn)動(dòng)物對(duì)T-2毒素的代謝顯得至關(guān)重要。之前T-2毒素的研究主要集中在哺乳動(dòng)物及家禽體內(nèi)的代謝研究,而在水產(chǎn)動(dòng)物中的研究較少。為此,深入研究對(duì)蝦對(duì)T-2毒素的清除能力及代謝產(chǎn)物顯得尤為重要。T-2毒素在動(dòng)物中的代謝研究主要分為體內(nèi)和體外,動(dòng)物體內(nèi)的研究主要是對(duì)T-2毒素在動(dòng)物體內(nèi)代謝后組織器官及排泄物中T-2毒素代謝產(chǎn)物的研究。而體外的研究主要運(yùn)用動(dòng)物的肝微粒體進(jìn)行代謝,由于肝臟是動(dòng)物的主要解毒器官,其中含有大量的解毒酶系,體外代謝的研究能為體內(nèi)研究提供一定的參考。本論文主要運(yùn)用對(duì)蝦和小鼠肝微粒體體外代謝T-2毒素。同時(shí)對(duì)對(duì)蝦和小鼠代謝T-2毒素的能力進(jìn)行比較,期望找到對(duì)蝦與小鼠代謝的差別,為進(jìn)一步研究T-2毒素在對(duì)蝦體內(nèi)的代謝提供指導(dǎo),也為水產(chǎn)飼料中T-2毒素限量標(biāo)準(zhǔn)的制定和風(fēng)險(xiǎn)評(píng)估提供參考。本論文分為四章內(nèi)容,第一章是通過文獻(xiàn)綜述來(lái)主要介紹真菌毒素及其在不同動(dòng)物體內(nèi)的代謝研究,主要是T-2毒素在哺乳動(dòng)物和家禽中的代謝。以及在植物中的代謝和動(dòng)物中的代謝差別。第二章主要是制備對(duì)蝦和小鼠肝微粒體,并檢測(cè)肝微粒體中的蛋白,用對(duì)硝基苯酚法檢測(cè)II相代謝酶UDPGT的活性。測(cè)得對(duì)蝦肝微粒體中的蛋白含量是14.9 mg/g,小鼠肝微粒體中的蛋白含量是10.1 mg/g。對(duì)蝦肝微粒體中的蛋白濃度高于小鼠。在對(duì)蝦肝微粒體中幾乎沒有檢測(cè)到UDPGT活性,而測(cè)得小鼠中的UDPGT酶活性為24.23 pmol/min-1mg-1。第三章是T-2-Glu A的制備及條件優(yōu)化。利用對(duì)蝦和小鼠肝微粒體制備T-2-GluA,但發(fā)現(xiàn)用對(duì)蝦肝微粒體未合成出產(chǎn)物,可能是由于對(duì)蝦和小鼠肝微粒體中所含的酶系的不同,可能是由于對(duì)蝦肝微粒體中不存在UDPGT酶。用小鼠肝微粒體體外酶法合成T-2-GluA,并對(duì)合成條件進(jìn)行了優(yōu)化,肝微粒體蛋白濃度越大合成量越大,孵育時(shí)間90 min,曲拉通濃度為0.2%(w/v)時(shí)合成量最大。第四章用對(duì)蝦和小鼠肝微粒體代謝T-2毒素,進(jìn)行體外研究,T-2毒素在不同動(dòng)物肝微粒體中的代謝存在明顯差異,小鼠肝微粒體對(duì)T-2毒素的清除力是對(duì)蝦肝微粒體的200多倍,說明對(duì)蝦感染T-2毒素后在體內(nèi)殘留的時(shí)間也越久,對(duì)對(duì)蝦造成的危害也越大。T-2毒素在對(duì)蝦和小鼠肝微粒體中的代謝產(chǎn)物也不同,這主要是肝微粒體中代謝酶系的不同,水產(chǎn)動(dòng)物與哺乳動(dòng)物的代謝的差異都需要進(jìn)一步研究。
[Abstract]:T-2 toxin is considered to be the most toxic in group A monotelene toxin, which has been used as a biological and chemical weapon in war because of its high toxicity. Some fungi can easily produce mycotoxins after they are infected with cereals under suitable conditions. Due to the increasing proportion of plant ingredients in aquatic feed, the harm of T-2 toxin to aquatic animals is also increasing, which can not only lead to the decline of quality of aquatic products, but also bring serious food safety problems. Therefore, it is very important to study the metabolism of T-2 toxin in aquatic animals. Previous studies of T-2 toxin were mainly focused on metabolism in mammals and poultry, but less in aquatic animals. Therefore, it is very important to study the scavenging ability and metabolites of T-2 toxin in prawn. The metabolism of T-2 toxin in animals is mainly divided into in vivo and in vitro. T-2 toxin metabolites in tissues, organs and excreta after metabolism of T-2 toxin in animals were studied in vivo. However, in vitro studies mainly use animal liver microsomes for metabolism. As the liver is the main detoxification organ of animals, which contains a large number of detoxification enzymes, the study of in vitro metabolism can provide a certain reference for in vivo research. In this paper, we used prawn and mouse liver microsome to metabolize T-2 toxin in vitro. At the same time, the ability of prawn and mice to metabolize T-2 toxin was compared, in order to find out the difference of metabolism between prawn and mouse, and to provide guidance for further study on metabolism of T-2 toxin in prawn. It also provides a reference for the formulation and risk assessment of the limit of T-2 toxin in aquatic feed. This paper is divided into four chapters. The first chapter is a review of the literature to introduce mycotoxins and their metabolism in different animals, mainly on the metabolism of T-2 toxin in mammals and poultry. And metabolic differences in plants and animals. In the second chapter, shrimp and mouse liver microsome were prepared, and the protein in liver microsome was detected. The activity of II metabolizing enzyme UDPGT was detected by p-nitrophenol method. The protein content in shrimp liver microsomes was determined to be 14. 9 mg/g, mouse liver microsomal protein content is 10. 1 mg/g.. The protein concentration in shrimp liver microsomes was higher than that in mice. Almost no UDPGT activity was detected in shrimp liver microsomes, while the activity of UDPGT enzyme in mice was 24.23 pmol/min-1mg-1.. The third chapter is about the preparation and optimization of T-2-Glu A. T-2-GluA was prepared from prawn and mouse liver microsomes, but it was found that the unsynthesized products from shrimp liver microsomes may be due to the different enzyme systems in shrimp and mouse liver microsomes. This may be due to the absence of UDPGT enzyme in shrimp liver microsomes. T-2-GluA was synthesized from mouse liver microsomes by enzymatic method in vitro, and the synthesis conditions were optimized. The higher the concentration of liver microsomal protein, the greater the amount of synthesis, and the maximum amount of synthesis was obtained when the incubation time was 90 min, when the concentration of Traitone was 0.2% (w / v). In chapter 4, the metabolism of T-2 toxin by prawn and mouse liver microsomes was studied in vitro. The metabolism of T-2 toxin in different animal liver microsomes was significantly different. The clearance of T-2 toxin by mouse liver microsomes was more than 200 times of that of prawn liver microsomes, which indicated that shrimp remained in vivo for a longer time after infection with T-2 toxin. The more harmful to prawns, the different metabolites of T-2 toxin in shrimp and mouse liver microsomes, which are mainly due to the different metabolic enzymes in liver microsomes. The differences in metabolism between aquatic animals and mammals require further study.
【學(xué)位授予單位】：廣東海洋大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S945.1;S859.8

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本文編號(hào)：2336140