本文選題：石油污染 + 生物修復(fù)�。� 參考：《青島理工大學(xué)》2014年碩士論文

【摘要】：石油是當(dāng)今社會(huì)生產(chǎn)發(fā)展的重要能源。隨著人類社會(huì)的快速進(jìn)步與發(fā)展，石油需求量大大增加，這就導(dǎo)致每年都有大量的石油由于突發(fā)事件進(jìn)入海洋等水域，造成極其嚴(yán)重的后果，對海洋生態(tài)平衡構(gòu)成了嚴(yán)重破壞與威脅。在石油發(fā)生泄漏后，往往在第一時(shí)間向海洋中投加分散劑，盡可能的降低石油污染對海洋的影響，并且在后續(xù)生物修復(fù)過程中會(huì)投加生物柴油促進(jìn)石油的生物降解，分散劑和生物柴油的投加都可能會(huì)對石油降解微生物造成影響。目前，對分散劑以及生物柴油處理后的石油污染海水中微生物種群多樣性的變化尚未開展系統(tǒng)的研究。 因此，本課題將基于分子生物學(xué)的克隆文庫技術(shù)和PCR-RFLP技術(shù)相結(jié)合的方法，研究并分析分散劑以及生物柴油的投加對石油污染海水中微生物群落結(jié)構(gòu)多樣性的影響，并在研究中得到以下結(jié)論： （1）隨著分散劑投加量的增加，石油污染海水中的優(yōu)勢菌群逐漸從食堿菌科Alcanivorax、海洋螺菌科Marinomonas、黃桿菌科Flavobacterium以及產(chǎn)堿菌科Achromobacter向紅菌科Parvibaculum、紅螺菌科Thalassospira過渡。 （2）隨著生物柴油投加量的增大，其污染海水中的優(yōu)勢菌群逐漸由食堿菌科Alcanivorax、紅菌科Parvibaculum和海洋螺菌科Marinomonas變?yōu)榧t螺菌科Thalassospira，鞘脂單胞菌科Novosphingobium。 （3）生物柴油的投加與分散劑相比更能促進(jìn)海水中微生物的生長，提高生物降解效率。分散劑以及生物柴油在相同的投加量下，投加生物柴油的污染海水中微生物的含量明顯高于投加分散劑的污染海水且石油降解率也是投加生物柴油的海水中較高。 （4）克隆子測序結(jié)果通過NCBI比對后顯示，16S rDNA克隆文庫中均只有50%-70%的克隆子測序結(jié)果可以在NCBI中找到相似度大于97%的菌株。因此表明，在各個(gè)條件下海水中石油降解微生物中均存在部分未被人類分類培養(yǎng)和鑒定的細(xì)菌種類。 （5）石油污染海水中，，海水細(xì)菌主要由變形菌綱以及黃桿菌綱和放線菌綱的細(xì)菌組成，其中α-變形菌綱以及γ-變形菌綱占絕對優(yōu)勢，分散劑的投加有利于γ-變形菌綱以及黃桿菌綱微生物的生長與繁殖，而生物柴油的投加更有利于α-變形菌綱微生物的生長繁殖。 （6）生物柴油以及分散劑的投加能促進(jìn)芳香烴降解菌大量繁殖，并逐步成為優(yōu)勢菌群，且隨著分散劑以及生物柴油投加量的增加，污染海水中芳香烴降解菌的比重也逐步增大，這表明生物柴油以及分散劑的投加能促進(jìn)石油中芳香烴類物質(zhì)的降解。
[Abstract]:Petroleum is an important energy source for the production and development of our society.With the rapid progress and development of human society, the demand for oil has increased greatly. This has led to a large amount of oil entering the sea and other waters every year due to unexpected events, resulting in extremely serious consequences.It is a serious damage and threat to the marine ecological balance.After oil spills, dispersants are often added to the ocean for the first time to minimize the impact of oil pollution on the ocean, and biodiesel is added to the subsequent bioremediation process to promote the biodegradation of oil.The addition of dispersants and biodiesel may have an impact on petroleum degradation microbes.At present, no systematic study has been carried out on the variation of microbial population diversity in seawater contaminated by petroleum and biodiesel treated with dispersants and biodiesel.Therefore, based on the methods of molecular biology and PCR-RFLP, the effects of dispersant and biodiesel on the diversity of microbial community in petroleum polluted seawater were studied and analyzed.The conclusions are as follows:1) with the increase of dispersant dosage, the dominant flora of petroleum polluted seawater gradually transitioned from Alcanivorax, Marinomonas, Flavobacterium and Achromobacter to Parvibaculum and Thalassospira.2) with the increase of biodiesel dosage, the dominant flora in polluted seawater changed from Alcanivorax, Parvibaculum and Marinomonas to Thalassospira and Novosphingobium.3) compared with dispersant, the addition of biodiesel can promote the growth of microorganisms in seawater and improve the biodegradation efficiency.Under the same dosage of dispersant and biodiesel, the microbial content of polluted seawater added with biodiesel was significantly higher than that of polluted seawater with dispersant, and the oil degradation rate was also higher in the seawater fed with biodiesel.The results of clone sequencing showed that only 50% -70% clones of 16s rDNA clones could be found in NCBI with a similarity of more than 97%.It is concluded that there are some species of petroleum degrading microorganisms in seawater which have not been cultured and identified by human taxonomy.In petroleum polluted seawater, seawater bacteria are mainly composed of Proteus, xanthobacterium and actinomycetes, among which 偽 -Proteus and 緯 -Proteus are dominant.The addition of dispersant is beneficial to the growth and reproduction of 緯 -Proteus and Xanthobacterium, while the addition of biodiesel is more favorable to the growth and reproduction of 偽 -Proteus.(6) the addition of biodiesel and dispersant can promote the proliferation of aromatic hydrocarbon degrading bacteria and gradually become the dominant flora, and with the increase of dispersant and biodiesel dosage, the proportion of aromatic hydrocarbon degrading bacteria in polluted sea water increases gradually.This indicates that the addition of biodiesel and dispersant can promote the degradation of aromatic hydrocarbons in petroleum.
【學(xué)位授予單位】：青島理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：X172;X55

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