本文選題：微生物傳感器　切入點(diǎn)：微生物膜　出處：《青島理工大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：生化需氧量(BOD)是表征水體中有機(jī)物等需氧污染物質(zhì)含量的重要指標(biāo)之一,現(xiàn)在普遍用于測(cè)定生物化學(xué)需氧量的標(biāo)準(zhǔn)方法是五日生化需氧量法,即樣品在20℃±1℃的溫度下培養(yǎng)5d,分別測(cè)定樣品培養(yǎng)前后的溶解氧,二者之差即為5d的生化需氧量。此方法是十分復(fù)雜的生物化學(xué)過(guò)程,受多種因素的影響,測(cè)試周期過(guò)于長(zhǎng),效率低,測(cè)試結(jié)果不穩(wěn)定,重現(xiàn)性差,所以方法本身存在很多的局限性。微生物傳感器快速測(cè)定法克服了上述測(cè)定法的缺點(diǎn),使整個(gè)測(cè)定周期大大縮短,測(cè)試結(jié)果直接打印出來(lái),方便快捷,并且可以實(shí)現(xiàn)在線監(jiān)測(cè),從而日益得到廣泛的應(yīng)用。本文闡述了微生物傳感器的產(chǎn)生、微生物傳感器的原理及組成、微生物傳感器的研究進(jìn)展包括微生物傳感器的類型、微生物的類型及固定化方法,對(duì)微生物傳感器的應(yīng)用BOD快速檢測(cè)儀進(jìn)行了簡(jiǎn)單介紹及運(yùn)用。重點(diǎn)對(duì)核心部分敏感材料做了研究,包括敏感材料微生物膜的研究現(xiàn)狀、制備過(guò)程中遇到的問(wèn)題及敏感材料微生物小球性能分析。本研究的重點(diǎn)為菌種的選擇、菌種培養(yǎng)溫度的選擇及固定化方法的選擇。1.本文敏感材料的制備首先選用傳統(tǒng)的生物膜法。選擇四個(gè)影響因素,分別是菌種、包埋劑、培養(yǎng)溫度、包埋劑與菌種配比,每個(gè)因素分為三個(gè)水平,即菌種:1#、2#、3#;溫度:25℃、30℃、35℃;包埋劑:聚乙烯醇、聚丙烯酰胺、海藻酸鈉;包埋劑與菌種配比:1:2、2:1、1:1。使用正交設(shè)計(jì)對(duì)這四因素三水平進(jìn)行實(shí)驗(yàn)安排,根據(jù)實(shí)驗(yàn)安排進(jìn)行微生物膜的制作,使用BOD快速檢測(cè)儀對(duì)標(biāo)準(zhǔn)溶液(BOD=2000±160mg/L)BOD值進(jìn)行測(cè)試。然后進(jìn)行正交分析,包括極差分析和方差分析。得出各因素對(duì)實(shí)驗(yàn)指標(biāo)的影響程度,按影響大小排序?yàn)?菌種、培養(yǎng)溫度、包埋劑、包埋劑與菌體配比。其中菌種的選擇和培養(yǎng)溫度對(duì)實(shí)驗(yàn)結(jié)果影響明顯,其余兩個(gè)次之。進(jìn)一步分析出來(lái)的最優(yōu)方案即選用菌種3#、培養(yǎng)溫度35℃、包埋劑聚乙烯醇、固定化材料與菌體配比1:2。最后對(duì)其進(jìn)行實(shí)驗(yàn)驗(yàn)證,分別對(duì)BOD值為2000±160mg/L的標(biāo)準(zhǔn)溶液及實(shí)際水樣進(jìn)行測(cè)定,證明最優(yōu)方案是符合要求的并對(duì)于不同來(lái)源水樣測(cè)定均具有較好的準(zhǔn)確性。在最優(yōu)方案所對(duì)應(yīng)的實(shí)驗(yàn)條件下,使用課題組的專利技術(shù),用微生物小球作為敏感材料,使用BOD快速檢測(cè)儀分別對(duì)BOD值為2000±160mg/L的標(biāo)準(zhǔn)溶液進(jìn)行測(cè)試并進(jìn)行數(shù)據(jù)分析,結(jié)果表明:相同的實(shí)驗(yàn)條件、實(shí)驗(yàn)材料下,使用微生物小球作為敏感材料的效果要好于使用微生物膜。數(shù)值上下波動(dòng)幅度更小,測(cè)試結(jié)果很穩(wěn)定,重現(xiàn)性好。用其測(cè)定的水樣的BOD值與5日培養(yǎng)法無(wú)顯著差異,這表明該生物小球?qū)τ诓煌瑏?lái)源水樣測(cè)定均具有較好的準(zhǔn)確性。篩選出最優(yōu)方案中對(duì)有機(jī)物降解能力強(qiáng)的3#菌種進(jìn)行菌種鑒定,首先革蘭氏染色,然后利用16S r DNA序列同源性分析技術(shù),進(jìn)行DNA提取、PCR擴(kuò)增后,送往上海桑尼生物科技有限公司完成測(cè)序反應(yīng)。菌種最終鑒定結(jié)果:細(xì)菌(Bacteria)厚壁菌門(mén)(Firmicutes)、芽孢桿菌綱(Bacilli)、芽孢桿菌目(Bacillales)、芽孢桿菌科(Bacillaceae)、芽孢桿菌屬(Bacillus.sp)。
[Abstract]:Biochemical oxygen demand (BOD) is one of the important indexes in water organic pollutants such as aerobic content, now commonly used standard method for the determination of BOD is Bod five, which were cultured in 5D 20 C + 1 DEG C temperature, dissolved oxygen samples were measured before and after training two, the difference is the biochemical oxygen demand 5D. This method is the biochemical process is very complex, affected by many factors, the test cycle is too long, low efficiency, unstable test results, poor reproducibility, so the method itself has many limitations. The rapid determination to overcome the above determination the shortcomings of microbial sensor, measurement period is greatly shortened, convenient test results directly printed, and can realize online monitoring, which has been widely used. This paper expounds the microbial sensor, micro The principle and composition of biosensor, research progress of microbial sensor types including microbial sensor, microorganism types and immobilization methods, the application of BOD rapid detection of microbial sensor are introduced and used. Focuses on the research on the core part of sensitive materials, including the research status of sensitive material of biofilm, analysis the preparation process and the problems in the sensitive materials for microbial ball performance. The focus of this study is the choice of culture, this paper select the.1. temperature sensitive materials and the selection of immobilized bacteria culture method in the preparation of the first selection of biofilm in the traditional method. Four factors are bacteria, entrapment agent, culture temperature, embedding agent and strain ratio, each factor is divided into three levels, namely: 1#, 2#, 3# strains; temperature: 25 C, 30 C, 35 C; embedding agent: polyvinyl alcohol, polyacrylamide, alginate Sodium; embedding agent and strain ratio: 1:2,2:1,1:1. using orthogonal design of the four factors and three levels of experimental arrangement, according to the experimental design and production of microbial membrane, the standard solution using BOD fast detector (BOD=2000 + 160mg/L) BOD values were tested. Then the orthogonal analysis, including the analysis of range and variance of. Various factors on the extent of the index, according to the influence of the order of strain, culture temperature, embedding agent, embedding agent and cell ratio. One species selection and cultivation of the temperature effect on the experimental results, the remaining two times. The optimal solution is selected out for further analysis of strain 3#, culture temperature 35 polyvinyl alcohol, embedding agent, immobilized cell material and the ratio of 1:2. finally carries out experiments on it, respectively. The value of BOD as the standard solution of 2000 + 160mg/L and the actual water samples were measured, it is proved that the optimal solution is To meet the requirements for different sources of water samples were measured and has good accuracy. The experimental conditions corresponding to the optimal scheme, using the research group of the patented technology, using microorganism balls as a sensitive material, the use of BOD rapid detection instrument respectively. The value of BOD was 2000 + 160mg/L Standard Test and data analysis, the quasi solution results show that the same experimental conditions, experimental materials, the use of microbial balls as better sensitive materials in the use of microbial membrane. The numerical fluctuation amplitude is smaller, the test result is stable, with good reproducibility. And the 5 day culture method had no significant difference with the determination of the BOD value of water sample, which indicates that the bio beads were has good accuracy for different sources of water samples were determined. Screened strains were identified on 3# strains of organic matter degradation ability of the optimal scheme, firstly, Gram staining, and then use 16S R Sequence analysis technology DNA, DNA extraction, PCR amplification, sent to Shanghai Sonny biological technology Limited completed the sequencing reaction. The final identification results: bacterial strains (Bacteria) Firmicutes bacilli (Firmicutes), (Bacilli), bacillales (Bacillales), Bacillaceae (Bacillaceae). The genus Bacillus (Bacillus.sp).

【學(xué)位授予單位】：青島理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：X853;TP212.3

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