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Population analysis of microbial consortia during oil degradation process at the in situ experiment field situated on a beach in Japan

日本海滩原位实验场石油降解过程中微生物群落的种群分析

基本信息

批准号：
14560150
负责人：
YOSHINAGA Ikuo
金额：
$ 2.3万
依托单位：
Graduate School of Agriculture, Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560150/
关键词：
bioremediation PCR-DGGE bacterial consortia marine bacteria biological additive oil degradation beach

项目摘要

We constructed an open-environment experiment field on a sandy beach facing to Mazuru Bay in Japan, for the purpose of verifying the efficiency of newly developed fertilizer Z18 (Japan Patent Office P2001-87754A) on bioremediation of crude oil pollution. In the experiment field, periodical tidal movement supplied oxygen and several nutrients for heterotrophic bacteria from seawater, and we placed several small permeable bags with artificially oil-contaminated sand samples at the intertidal depth. On experimental days 7, 14, 28, 56 and 112, we analyzed the oil degradability, the number of oil-degrading bacteria (ODB) and bacterial population in each sand bag, and compared them between the fertilized and non-fertilized samples. The ODB numbers were determined by a MPN method and bacterial population was estimated by a culture-independent protocol, PCR-DGGE (denaturing gradient gel electrophoresis) targeting bacterial 16S rRNA genes. The nitrogen containing fertilizer Z18 excluded oil com … More ponents (saturated alkanes, aromatics, resins and asphaltenes) more effectively than non-fertilized samples. The growth of GOB and the biodegradability of oil (estimated from the survivability of normal alkanes relative to hopane) were also accelerated by Z18 addition. Contrarily, the bacterial population dynamics estimated by PCR-DGGE patterns during the oil-degrading process were not greatly different between fermented and non-fermented samples, suggesting that the fermentation of Z18 did not promote a particular ODB denitrifying bacteria belonging to alpha-proteobacteria subgroup. Unfortunately, dissolved inorganic nitrogen (DIN) had gradually accumulated in fertilized samples after 56 days of experiment, by which time almost normal alkanes were diminished, and there remained the problem of eutrophication of marine environments. However, we compared the effect of oil degradation and accumulation of DIN among the samples to which various forms, and concentrations of Z18 applied, and proposed an ingenious application of the new fertilizer for spilt oil treatment on seashore, without excess nitrogen accumulation. Less

为了验证新开发的肥料Z18（日本专利局P2001- 87754 A）对原油污染的生物修复的效率，我们在日本马祖湾（Mazuru Bay）对面的桑迪海滩上构建了开放环境实验场。在实验场，周期性的潮汐运动为海水中的异养细菌提供氧气和多种营养物质，我们在潮间深度放置了几个装有人工污染石油的砂样的小渗透袋。在试验的第7、14、28、56和112天，分析了每个砂袋中的石油降解率、石油降解菌（ODB）数量和细菌种群数量，并对施肥和未施肥样品进行了比较。通过MPN方法测定ODB数，并通过针对细菌16 S rRNA基因的不依赖于培养的方案PCR-DGGE（变性梯度凝胶电泳）估计细菌种群。Z18脱油专用氮肥 ...更多信息与未施肥的样品相比，更有效地去除有机质（饱和烷烃、芳香族化合物、树脂和沥青质）。GOB的增长和石油的生物降解性（估计从生存的正构烷烃相对于藿烷）也加快了Z18的加入。通过PCR-DGGE图谱分析发现，发酵和未发酵样品在油脂降解过程中的细菌种群动态没有显著差异，表明Z18的发酵并没有促进一种属于α-变形菌亚群的特定ODB降解细菌。不幸的是，溶解无机氮（DIN）逐渐积累在施肥样品后56天的实验，到那时几乎正构烷烃减少，仍然存在的问题，海洋环境的富营养化。然而，我们比较了不同形式和浓度的Z18施用的样品之间的油降解和DIN积累的效果，并提出了一种巧妙的新肥料的应用，用于在海滨处理溢油，而没有过量的氮积累。少