Evans Flavia F, Rosado Alexandre S, Sebastián Gina V, Casella Renata, Machado Pedro L O A, Holmström Carola, Kjelleberg Staffan, Elsas Jan D, Seldin Lucy
Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro (UFRJ), CCS, Bloco I, Ilha do Fundão, Rio de Janeiro, RJ, CEP: 21941-590, Brazil.
FEMS Microbiol Ecol. 2004 Aug 1;49(2):295-305. doi: 10.1016/j.femsec.2004.04.007.
The aim of this study was to analyse the effect of oil contamination and biostimulation (soil pH raise, and nitrogen, phosphate and sulphur addition) on the diversity of a bacterial community of an acidic Cambisol under Atlantic Forest. The experiment was based on the enumeration of bacterial populations and hydrocarbon degraders in microcosms through the use of conventional plating techniques and molecular fingerprinting of samples directly from the environment. PCR followed by denaturing gradient gel electrophoresis (DGGE) was used to generate microbial community fingerprints employing 16S rRNA gene as molecular marker. Biostimulation led to increases of soil pH (to 7.0) and of the levels of phosphorus and K, Ca, and Mg. Oil contamination caused an increase in soil organic carbon (170-190% higher than control soil). Total bacterial counts were stable throughout the experiment, while MPN counts of hydrocarbon degraders showed an increase in the biostimulated and oil-contaminated soil samples. Molecular fingerprinting performed with 16S rRNA gene PCR and DGGE analysis revealed stable patterns along the 360 days of experiment, showing little change in oil-contaminated microcosms after 90 days. The DGGE patterns of the biostimulated samples showed severe changes due to decreases in the number of bands as compared to the control samples as from 15 days after addition of nutrients to the soil. Results obtained in the present study indicate that the addition of inorganic compounds to soil in conjunction with oil contamination has a greater impact on the bacterial community than oil contamination only.
本研究的目的是分析油污污染和生物刺激（提高土壤pH值，添加氮、磷和硫）对大西洋森林下酸性始成土细菌群落多样性的影响。该实验基于通过使用传统平板技术对微宇宙中的细菌种群和烃降解菌进行计数，并对直接从环境中采集的样品进行分子指纹分析。采用聚合酶链反应（PCR）后进行变性梯度凝胶电泳（DGGE），以16S rRNA基因作为分子标记来生成微生物群落指纹。生物刺激导致土壤pH值升高至7.0，磷以及钾、钙和镁的含量增加。油污污染使土壤有机碳增加（比对照土壤高170 - 190%）。在整个实验过程中，细菌总数保持稳定，而在经过生物刺激和油污污染的土壤样品中，烃降解菌的最大可能数（MPN）计数有所增加。用16S rRNA基因PCR和DGGE分析进行的分子指纹分析显示，在360天的实验过程中模式稳定，在90天后油污污染的微宇宙中变化很小。与对照样品相比，从向土壤中添加养分后15天起，生物刺激样品的DGGE模式因条带数量减少而显示出严重变化。本研究获得的结果表明，向土壤中添加无机化合物并结合油污污染对细菌群落的影响比仅油污污染更大。
