利用16S rDNA测序测定尼日利亚伊贝诺偏远航空燃料污染生态系统的原核生物多样性

I. Udotong, M. Uko, J. Udotong
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摘要

Ibeno是埃克森美孚的子公司Mobil production Nigeria Unlimited的运营基地,是尼日利亚尼日尔三角洲地区受油气活动影响最大的地区之一。由于石油泄漏和油类废物的排放以及加油活动,该地区居民所依赖的油气系统几乎每天都要接收石油烃的输入。本研究采用宏基因组方法确定了一个偏远的航空燃料污染的生物生态系统经过16年污染后的原核生物多样性。按照制造商的说明,使用MoBio DNA提取试剂盒从水样中提取DNA。提取的DNA片段用picogreen定量,用NanoDrop分光光度计记录其紫外吸收光谱。16S rDNA测序在Miseq Illumina测序平台和QIIME生物信息学管道上进行。分析表明,在污染和未污染的水样中,细菌和古细菌群落都占主导地位。污染样品细菌总数为93.83%,古菌总数为3.43%;对照样品细菌总数为58.05%,古菌总数为39.69%。污染样品的优势菌门为变形菌门、厚壁菌门、放线菌门、蓝藻门和氯霉素门,未污染样品的优势菌门为变形菌门、厚壁菌门和放线菌门。污染水体和未污染水体的优势古菌门类均为Euryarchaeota和Crenarchaeota。利用16S rDNA宏基因组方法揭示了污染点和对照点细菌和古细菌的多样性,从而揭示了污染点和对照点的真实生态状况。
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Use of 16S rDNA Sequencing to Determine Procaryotic Diversity of a Remote Aviation Fuel-Polluted Lentic Ecosystem in Ibeno, Nigeria
Ibeno, the operational base of Mobil Producing Nigeria Unlimited, a subsidiary of ExxonMobil, Nigeria remains one of the most impacted communities by oil and gas activities in the Niger Delta region of Nigeria. Lotic and lentic systems in the region which residents rely on, receive petroleum hydrocarbon inputs almost daily due to oil spills and oily wastes discharges from operators and bunkering activities. This research was carried out to determine the prokaryotic diversity in a remote aviation fuel-polluted lentic ecosystem after 16 years of pollution using metagenomic approaches. DNA extraction from the water samples was carried out using MoBio DNA extraction Kits following the manufacturer’s instructions. Extracted DNA fragments were quantified using picogreen and by recording their UV absorption spectra using NanoDrop spectrophotometer. 16S rDNA sequencing was carried out on a Miseq Illumina sequencing platform and Quantitative Insight Into Microbial Ecology (QIIME) bioinformatics pipeline. Analyses revealed the dominance of bacterial and archaeal communities in both polluted and unpolluted water samples. The polluted sample had 93.83% bacterial and 3.43% archaeal population while the control sample revealed 58.05% bacterial and 39.69% archaeal population. Dominant bacterial phyla from the polluted samples were Proteobacteria, Firmicutes, Actinobacteria, Cyanobacteria, and Chloroflexi while dominant phyla in the unpolluted samples were Proteobacteria, Firmicutes and Actinobacteria. Dominant archaeal phyla from both polluted and unpolluted waters were Euryarchaeota and Crenarchaeota. The use of 16S rDNA metagenomic approach revealed a wide variety of bacterial and archaeal diversity from both polluted and control sites, thus revealing the true ecological status of both sites.
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