本文選題：溫和氣單胞菌 + 強(qiáng)弱毒株��； 參考：《吉林農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：溫和氣單胞菌在自然界中分布廣泛,是人獸共患的機(jī)會性致病菌。當(dāng)機(jī)體免疫力低下時,溫和氣單胞菌易從潛伏菌變?yōu)橹虏【?并且使用大量的抗菌素易使菌株變?yōu)槟退幘�。本研究從病死魚中分離出2株溫和氣單胞菌,通過動物回歸試驗(yàn)檢測其毒株強(qiáng)弱;通過Illumina HiSeq 2500測序儀將提取DNA構(gòu)建的文庫進(jìn)行全基因組重測序,并利用COG、GO數(shù)據(jù)庫對比分析2株菌的基因差異;通過全基因組重測序?qū)ふ褻RISPR位點(diǎn)并進(jìn)行耐藥機(jī)制檢測;通過5種常用水產(chǎn)藥物檢測2株溫和氣單胞菌體外抑菌濃度并確定最佳抑菌時間。為溫和氣單胞菌的致病機(jī)制及耐藥機(jī)制提供科學(xué)依據(jù)。其實(shí)驗(yàn)結(jié)果如下:1動物回歸試驗(yàn)檢測結(jié)果為,A1菌半致死濃度為2.7×105 cfu/0.5 mL;B2菌半致死濃度為1.6×105 cfu/0.5 mL。2 A1、B2蛋白編碼區(qū)域的SNP中,77%是同義編碼突變,23%非同義編碼突變。3 2株菌株中有24293個核苷酸為非同義突變。A1發(fā)生的非同義突變基因數(shù)目為2492,B2發(fā)生的非同義突變基因數(shù)目為2488。4 A1、B2所產(chǎn)生的差異基因突變個數(shù)分別為2564、2565。5 B2突變基因位置存在于gene rna1865位,這也使得在統(tǒng)計(jì)COG數(shù)據(jù)時,碳水化合物轉(zhuǎn)運(yùn)與代謝[G],A1為144個,B2為145個。6編碼rna 1865位基因的堿基一共有135個,一共有107個SNPs基因位點(diǎn)發(fā)生突變,有1個基因發(fā)生非同義編碼突變。7與參考基因組比較,在參考序列位點(diǎn)坐標(biāo)第27407,T堿基突變?yōu)锳堿基。8 2株溫和氣單胞菌中可信CRISPR個數(shù)為1,總長度為2805bp;42條間隔序列;可疑CRISPR個數(shù)為3,總長度為384bp。對可信CRISPR的重復(fù)序列二級結(jié)構(gòu)進(jìn)行預(yù)測表明,頭環(huán)由15個堿基構(gòu)成;莖環(huán)含有5個堿基,總體形成發(fā)夾環(huán)。在進(jìn)行BLAST比對時顯示,除其他同屬或同源種屬外,DR與嗜熱螺旋體DSM 6192、暗網(wǎng)菌屬等遠(yuǎn)緣菌種均有較高的同源性,且置信值為1。9 5種常用水產(chǎn)藥物對2株溫和氣單胞菌均能產(chǎn)生PAE,8 x MIC的作用時間最短、16 x MIC和32 x MIC作用的時間最長。這均與已研究的結(jié)果相符。并且低濃度的服樂興和恩諾殺星均能對產(chǎn)生較長的PAE。
[Abstract]:Aeromonas mildii, which is widely distributed in nature, is the opportunistic pathogen of zoonosis. When immunity is low, Aeromonas mildii can easily change from latent bacteria to pathogenic bacteria, and the use of a large number of antibiotics can easily turn the strains into drug-resistant bacteria. In this study, two strains of Aeromonas mildifolia were isolated from diseased and dead fish, and their virulence was detected by animal regression test, and the library constructed by Illumina HiSeq 2500 was sequenced by Illumina HiSeq 2500 sequencer. The gene differences of the two strains were compared with COGG go database, and the CRISPR loci were found by genome re-sequencing and the drug resistance mechanism was detected. In vitro inhibitory concentration of 2 strains of Aeromonas mildii was detected by 5 kinds of commonly used aquatic drugs and the optimum bacteriostatic time was determined. It provides scientific basis for pathogenic mechanism and drug resistance mechanism of Aeromonas mildii. The results of the experiment are as follows: the result of the animal regression test is as follows: the semi-lethal concentration of A1A1 is 2.7 脳 105mLLB2, and the semi-lethal concentration of SNPs is 1.6 脳 105mL.2 A1OB2 protein coding region. 77% of the SNPs are synonymous coding mutations and 23.3% non-synonymous coding mutations. 2 strains of SNPs. There were 24293 nonsynonymous mutation genes with 2 4293 nucleotide nonsynonymous mutations. The number of non-synonymous mutations was 2492B 2. The number of non-synonymous mutation genes was 2488.4 A1nb 2. The number of differential gene mutations was 2564% 2565.552 B 2 mutation in the gene rna1865 locus, respectively. This also resulted in a total of 135 bases encoding 145 rna 1865 position genes in carbohydrate transport and metabolism [G] A 1 = 144 B 2 and 145 rna 1865 position genes, and 107 SNPs gene loci were mutated. Compared with the reference genome, the number of credible CRISPR was 1 and the total length of CRISPR was 2 805bp2 in the reference sequence coordinate 27407 T base mutation to A base 8.82 strain. The number of suspected CRISPR was 3 and the total length was 384bp. The secondary structure of the repeat sequence of trusted CRISPR was predicted. The head ring was composed of 15 bases, and the stem ring contained 5 bases, forming a hairpin ring as a whole. The comparison of blast showed that there was a high homology between Dr and DSM61922, except for other genus or homologous species, and the other distant species, such as the genus DSM61922, had a high homology with the genus DSM61922, and there was a high homology between Dr and DSM61922. The confidence value was that 1.95 common aquatic drugs could produce PAE 8 x MIC with the shortest time of 16 x MIC and 32 x MIC. These results are in good agreement with the results obtained. And low concentration of Lexing and enroxide can both produce a longer PAE.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S941.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 巴翠玉;張雅斌;張培軍;張蕾;冷東澤;李月紅;;大麻哈魚溫和氣單胞菌的分離鑒定及藥敏試驗(yàn)[J];中國獸醫(yī)科學(xué);2016年06期

2 商寶娣;張效平;;氣單胞菌性魚病研究進(jìn)展[J];農(nóng)業(yè)與技術(shù);2015年24期

3 耿昕穎;葛錚;孔令聰;朱世馨;單曉楓;馬紅霞;高云航;;52株不同淡水魚類維氏氣單胞菌耐藥表型的分析[J];中國獸藥雜志;2015年06期

4 王海娟;王利;;溫和氣單胞菌毒力基因的檢測及其對鯽魚致病性試驗(yàn)[J];動物醫(yī)學(xué)進(jìn)展;2015年03期

5 李莉;曹延超;謝旭陽;陳穎;戴文婷;孔祥會;;乳酸恩諾沙星對溫和氣單胞菌的體外抗菌后效應(yīng)[J];動物醫(yī)學(xué)進(jìn)展;2015年01期

6 黃鈞;彭民毅;羅華平;胡大勝;黃艷華;溫華成;施金谷;彭亞;;黃顙魚源溫和氣單胞菌對氟苯尼考耐藥性獲得與消失速率研究[J];西南農(nóng)業(yè)學(xué)報(bào);2013年03期

7 張利;李玉平;黎曉敏;尼瑪倉木拉;;牛支原體藥物敏感性試驗(yàn)[J];動物醫(yī)學(xué)進(jìn)展;2012年02期

8 黃冠軍;劉天強(qiáng);劉衍鵬;饒朝龍;肖丹;;溫和氣單胞菌PCR快速診斷方法的建立[J];淡水漁業(yè);2012年01期

9 張曉峰;楊晶;孫效文;;基于EST序列的鯉魚生長相關(guān)SNP發(fā)掘[J];水產(chǎn)學(xué)雜志;2009年04期

10 劉福平;白俊杰;葉星;李勝杰;李小慧;于凌云;;羅非魚MC4R基因克隆及與其生長相關(guān)的SNPs位點(diǎn)[J];中國水產(chǎn)科學(xué);2009年06期

相關(guān)碩士學(xué)位論文 前1條

1 胡攀;豬鏈球菌全基因組測序與比較基因組學(xué)研究[D];華中農(nóng)業(yè)大學(xué);2011年

，

本文編號：2042387