發(fā)布時間：2018-05-06 12:38

  本文選題：腸道微生物 + 微囊藻毒素　； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：隨著水產(chǎn)養(yǎng)殖集約化的發(fā)展,水體的富營養(yǎng)化越來越嚴(yán)重,藍(lán)藻水華頻發(fā)。藍(lán)藻水華的發(fā)生通常伴隨著藻毒素的產(chǎn)生,對魚類的生存和人類的安全造成了嚴(yán)重威脅。生物浮床因其環(huán)保實(shí)用,被廣泛應(yīng)用于降低水體富營養(yǎng)化水平,控制藍(lán)藻水華的發(fā)生。而上述過程中魚類腸道微生物是如何變化的,對機(jī)體起到了什么作用目前還不清楚。本研究以3月齡的性成熟斑馬魚為實(shí)驗(yàn)材料,對其進(jìn)行為期30 d的MC-LR的體外暴露,處理濃度分別為0、1、5、20μg/L,通過對暴露后斑馬魚腸道微生物16S rRNA基因測序,并測定其對MC-LR的吸收富集,超氧化物歧化酶(SOD),過氧化氫酶(CAT)和乳酸脫氫酶(LDH)活性、有機(jī)陰離子轉(zhuǎn)運(yùn)多肽Oatp2b1的表達(dá),而后作相關(guān)性分析,評價了腸道微生物在其中發(fā)揮的作用。同時,通過PCR-DGGE技術(shù)評價了三種植物浮床作對草魚腸道微生物和水體微生物的影響。具體結(jié)果如下:在用0、1、5、20μg/L的MC-LR處理30天后,斑馬腸道微生物α和β多樣性沒有產(chǎn)生顯著性差異,表明存在一個核心菌群,其由Fusobacteria,Actinobacteria,Alphaproteobacteria,Betaproteobacteria,Gammaproteobacteria等組成。然而,db-RDA分析顯示高劑量的MC-LR改變了高豐度微生物群的特征。腸道吸收MC-LR具有劑量依賴效應(yīng)。Oatp2b1表達(dá)隨著MC-LR劑量的增加而增加,SOD、CAT、LDH的活性在低劑量下顯著增加,而在較高劑量下則下降。使用線性判別分析效應(yīng)(LEfSe)和斯皮爾曼相關(guān)性檢驗(yàn)進(jìn)一步分析這些參數(shù)。鑒定的表型包括放線菌,乳桿菌和一些條件性致病菌。MC-LR暴露增加了致病菌的致病風(fēng)險,但腸道中的益生菌會通過競爭來抑制這種趨勢。浮床實(shí)驗(yàn)表明,不同植物根系細(xì)菌組成中沒有顯著差異,生物浮床對池塘水的微生物群落的豐度和優(yōu)勢菌群都沒有顯著影響。草魚腸道中的優(yōu)勢微生物從氣單胞菌Aeromonas jandaei變?yōu)轭愌挎邨U菌Paenibacillus sp.,證明生物浮床能夠調(diào)節(jié)腸道微生物群落結(jié)構(gòu)。
[Abstract]:With the development of intensive aquaculture, eutrophication of water is becoming more and more serious, and cyanobacteria Shui Hua frequently occurs. The occurrence of cyanobacteria Shui Hua is usually accompanied by the production of algal toxins, which poses a serious threat to the survival of fish and the safety of human beings. Biological floating bed is widely used to reduce eutrophication level and control the occurrence of cyanobacteria Shui Hua because of its environmental protection and practicality. It is not clear how the intestinal microbes change during the process, and what role they play in the organism. In this study, adult zebrafish (3 months old) were exposed to MC-LR for 30 days in vitro. The concentration of rRNA was 20 渭 g / L, respectively. The 16s rRNA gene of intestinal microorganism of zebrafish was sequenced, and the absorption and enrichment of MC-LR were determined. The activities of superoxide dismutase (SOD), catalase (catalase) and lactate dehydrogenase (LDH) and the expression of organic anion transport polypeptide (Oatp2b1) were studied. At the same time, the effects of three kinds of plant floating bed on intestinal microorganism and microorganism in water of grass carp were evaluated by PCR-DGGE technique. The results were as follows: after 30 days of treatment with 20 渭 g / L MC-LR, there was no significant difference between 偽 and 尾 diversity of intestinal microbes in zebra, indicating the existence of a core flora composed of Fusobacteria actinobacteriae Alphaproteobacteria Alpha-proteobacteriae Beta proteobacteria Gammaobacteria and so on. However, Db-RDA analysis showed that high dose of MC-LR changed the characteristics of high abundance microbial community. Intestinal absorption of MC-LR was dose-dependent. The expression of Oatp2b1 increased with the increase of MC-LR dose, and increased significantly at low dose, but decreased at higher dose. Linear discriminant analysis (LDA) and Spelman correlation test were used to further analyze these parameters. The identified phenotypes include actinomycetes lactobacillus and some conditioned pathogens. MC-LR exposure increases the risk of pathogenicity but the intestinal probiotics inhibit this trend through competition. The floating bed experiment showed that there was no significant difference in the composition of root bacteria among different plants, but the biological floating bed had no significant effect on the abundance and dominant flora of microbial community in pond water. The dominant microorganism in the intestine of grass carp changed from Aeromonas jandaei to Paenibacillus sp., which proved that the floating bed could regulate the structure of intestinal microbial community.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：X503.225;S917.4

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