Isolation and characterization of bacteria from soil contaminated with diesel oil and the possible use of these in autochthonous bioaugmentation

Two bacterial species (isolates N and O) were isolated from a paddy soil microcosm that had been artificially contaminated with diesel oil to which extrinsic Pseudomonas aeruginosa strain WatG, had been added exogenously. One bacterial species (isolate J) was isolated from a similar soil microcosm that had been biostimulated with Luria–Bertani (LB) medium. Isolates N and O, which were tentatively identified as Stenotrophomonas sp. and Ochromonas sp., respectively, by sequencing of their 16 S rRNA genes had no ability to degrade diesel oil on their own in any liquid medium. When each strain was cocultivated with P. aeruginosa strain WatG in liquid mineral salts medium (MSM) containing 1% diesel oil, isolate N enhanced the degradation of diesel oil by P. aeruginosa strain WatG, but isolate O inhibited it. In contrast, isolate J, which was tentatively identified as a Rhodococcus sp., degraded diesel oil contained not only in liquid LB and MSM, but also in paddy soil microcosms supplemented with LB medium. The bioaugmentation capacity of isolate J in soil microcosms contaminated with diesel oil was much higher than that of P. aeruginosa strain WatG. The possibility of using isolate J for autochthonous bioaugmentation is discussed.     

从两类不同培养基上分离的土壤细菌的特征比较

通过对两类不同培养基上分离的土壤细菌的生理特征的比较,发现两组细菌的种群有所不同.芽孢杆菌主要生长在宫营养的培养基上,而很难在低营养的培养基上生长.假单孢杆菌和棒杆类型的细菌则在两类培养基上均可生长.这与两类细菌对环境的适应能力有关.     

A novel method for sampling bacteria on plant root and soil surfaces at the microhabitat scale

This study reports the first method for sampling bacteria at a spatial scale approximating a microhabitat. At the core of this method is the use of tungsten rods with laser-cut tips of known surface area (0.013 mm(2)). Exposed plant root or soil surfaces were viewed with a dissecting microscope and micro-sampling rods were guided to sample sites using a micro-manipulator. Bacteria that adhered to the sampling tips were then recovered for microbiological analyses. The efficiency of this method for removing bacteria from root surfaces was similar to that with which bacteria are recovered from dissected root segments using the conventional technique of washing. However, as the surface area of the micro-sampling tips was known, the new method has the advantage of eliminating inaccuracy in estimates of bacterial densities due to inaccurate estimation of the root or soil surface sampled. When used to investigate spatial distributions of rhizoplane bacteria, the new technique revealed trends that were consistent with those reported with existing methods, while providing access to additional information about community structure at a much smaller spatial scale. The spatial scale of this new method is ca. 1000-times smaller than other sampling methods involving swabbing. This novel technique represents an important methodological step facilitating microbial ecological investigations at a microhabitat scale.     

Antibacterial activity of soil bacteria isolated from Kochi,India and their molecular identification

The present study, deal about the antibiosis activity of soil bacteria, isolated from 10 different locations of rhizosphere and diverse cultivation at Kochi, Kerala, India. The bacteria were isolated by standard serial dilution plate techniques. Morphological characterization of the isolate was done by Gram’s staining and found that all of them gram positive. Isolated bacteria were tested against 6 human pathogens viz., Escherichia coli, Enterococcus sp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus and Acinetobacter sp. Primary screening was carried out by perpendicular streaking and seed overlay method. Based on the result of primary screening most potential isolates of S1A1 and S7A3 were selected for secondary screening. Both the isolates showed positive results against Enterococcus sp. and S.aureus. The maximum antagonistic activity of 20.98 and 27.08?mm zone of inhibition was recorded at S1A1 against Enterococcus sp. and S. aureus respectively, at 180?µl concentration. Molecular identification was carried out by 16S rRNA sequence. The 16S rRNA was amplified from the DNA samples by using PCR. The amplified 16S rRNA PCR products were purified and sequenced. The sequences were subjected to NCBI BLAST. The isolates S1A1 and S7A3 BLAST results showed 99% and 95% respectively, similarity with the available database sequence of Bacillus amyloliquefaciens. The sequences were deposited in GenBank and the accession numbers KY864390 (S1A1) and KY880975 (S7A3) were obtained.     

海南岛不同地形上某些土壤化学性质的多样性分析

多样性指数长时间以来在生态学领域被用于分析不同生态体的发生规律．将类似手段用于某些土壤化学性质(土壤pH值、表层阳离子代换量和表层有机质含量)多样性的分析中,讨论了基于海南岛土壤一地体数字化数据库(SOTER),通过对不同地形上土壤pH值、表层阳离子代换量和表层有机质含量类别SOTER单元的丰富度指数(S)、多样性指数(Shannon指数H’)和均匀度指数(Pielou E)的计算,建立了类别多度分布模型,并在地理信息系统环境中实现了海南岛不同地形上土壤化学性质多样性分析的数字化表达．研究表明,上述研究路线具有可操作性。几种多样性指数和多度分布模型的建立可以较好地反映不同地形上土壤化学性质类别的空间变异特点．因此,SOTER单元作为土壤性质多样性的测度单元是可行的．     

艾比湖湿地氨氧化细菌数量空间分布及其与土壤环境相关性分析

【目的】对新疆艾比湖湿地不同植被类型(柽柳群落、盐节木群落、芦苇群落)和土壤深度(0-5 cm、5 cm-15 cm、15 cm-25 cm、25 cm-35 cm)中氨氧化细菌数量空间分布进行研究,并对其与土壤环境因子的相互关系进行分析。【方法】采用MPN-Griess和Pearson相关分析法。【结果】艾比湖湿地不同植被类型氨氧化细菌的数量存在明显的差异,分布趋势为柽柳群落最高,盐节木群落次之,芦苇群落最低;不同土层中氨氧化细菌的数量也存在明显的差异,分布趋势为15 cm-25 cm>0-5 cm>5 cm-15 cm>25 cm-35 cm;氨氧化细菌数量分布与土壤有机质含量呈显著相关,与土壤pH、含水量、盐度以及氨氮含量等因子之间均无相关性。【结论】艾比湖湿地不同植被类型和不同土层中氨氧化细菌数量的分布均存在显著差异;氨氧化细菌数量的空间分布除与土壤有机质含量呈显著相关外,与其他土壤环境因子均无相关性。     

Isolation and characterization of indigenous soil bacteria for bioaugmentation of PAH contaminated soil of semiarid Patagonia, Argentina

The aim of this work was to isolate PAH degrading-bacteria from contaminated Patagonia soil with the ability to tolerate the usual environmental stresses (oligotrophic and dryness conditions). Two approaches were utilized to obtain PAH-degrading bacteria from the Patagonian soil. With a traditional enrichment approach only the PAH- degrading strain 36 was isolated. Using a direct isolation approach three PAH-degrading strains (1A, 22A and 22B) were isolated. The phylogenetic analysis revealed that all isolates belonged to Sphingomonas genus. The PAH degrading activity and the resistance to stress conditions of the strains were determined and compared with those of the exogenous PAH-degrading Sphingomonas paucimobilis 20006FA. The strains 1A, 22A and 36 were phylogenetically closely related between them and with the strain 20006FA. The strain 22B, that showed a different phylogenetic position, was more resistant to C-starvation and drying conditions than other Patagonian strains. The effect of the inoculation of these strains on phenanthrene-induced mineralization and elimination was studied in Patagonian soil artificially contaminated, at different environmental conditions. The results suggest that strain 22B is the most suitable strain for bioaugmentation in PAH-contaminated soils of Central Patagonia, due to its adaptation to the usual environmental conditions. Our results show the importance of a detailed physiological characterization of isolates for autochthonous bioaugmentation strategies success.