The Isolation and Identification of the Indigenous Bacteria from Deep Subsurface Rocks of Talu Sandstone in the Miaoli Area

• Main Authors: Chung-Liang Hsieh, 謝崇良
• Other Authors: Jiin-Shuh Jean
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/98167387004558964225

碩士 === 國立成功大學 === 地球科學系碩博士班 === 91 === The study of deep subsurface bacteria is mainly from the deep subsurface rocks or geological formation water while drilling a well. The environments where the deep subsurface bacteria may be present include oil-bearing rocks, geological formation water, lacustrine deposits and sedimentary rock, hot spring, etc. However, only oil-bearing rocks in the Miaoli area were taken for this study. The samples of this study taken from the 3385-meter-deep subsurface rocks (i.e. Talu Sandstone) and drilling muds were collected in a certain drilling well in the Miaoli area. The indigenous bacteria from these rocks and muds are isolated and enriched with the MSA medium for the ferric-reducing bacteria and the TYG medium for the fermentative bacteria. The bacteria were identified followed by using the molecular biotechnology such as DNA extraction and purification, PCR (polymerase chain reaction), gel electrophoresis, gene sequencing and phylogenetic tree. Meanwhile, the laboratory tests for the ranges of temperature and salinity that the indigenous bacteria were adapted to the environment were performed. This is to understand the most optimal ranges for bacterial growth and to speculate the possible source of the indigenous bacteria. The electron microscopes were used to observe the morphology of bacteria. The experimental results of the study showed that the viable bacterial strains could obtain energy and grow by utilizing the reaction products of the fermentative and ferric-reducing bacteria. The sedimentation tests and the enrichment of bacterial culture from the ferric-reducing medium showed that there exist magnetic dark materials. The transformation of the reddish-brown Fe(Ⅲ)-oxyhydroxide in the ferric-reducing medium accounts for resulting from the bacterial reaction rather than from the chemical reaction. The isolated and identified strain 30H1L has 97﹪ similarities in nucleotide to Bacillus pumilus and Bacillus sp. PB10. It can be defined that strain 30H1L belongs to genus Bacillus.