Optimized extraction of polycyclic aromatic hydrocarbons from contaminated soil samples

A comparison of Soxhlet extraction and a new extraction technique, fluidized-bed extraction, has been conducted. The extraction of polycyclic aromatic hydrocarbons (PAHs) by this new technique has been optimized considering as experimental variables the variation of the number of extraction cycles and the holding time after reaching the heating temperature by means of a surface response design. The significance of the operational parameters of the fluidized-bed extraction onto the performance characteristics has been investigated. For the determination of the analytes, a cleanup of the extracts followed by gas chromatography with mass spectrometric detection was used. The accuracy of the method was established by extraction and analysis of a reference material, supplied from the European Commission's Joint Research Centre.     

Biodegradation of lindane by a native bacterial consortium isolated from contaminated river sediment

The aerobic biodegradation of lindane (γ-hexachlorocyclohexane) by a consortium of acclimated bacteria from sediment at a polluted site on the Suquia River, Cordoba, Argentina, is reported. The bacteria were acclimated for 30 days under aerobic conditions, using a minimal culture medium containing lindane (0.034 mM) as sole carbon source. Growth of the bacterial consortium decreased at a lindane concentration of 1.03 mM and was totally inhibited at 2.41 mM. The consortium showed initial lindane degradation rates of 4.92×10⁻³, 11.0×10⁻³ and 34.8×10⁻³ mM h⁻¹ when exposed to lindane concentrations of 0.069, 0.137 and 0.412 mM, respectively. Chloride concentration increased during aerobic biodegradation, indicating lindane mineralization. A metabolite identified as γ-2,3,4,5,6-pentachlorocyclohexene appeared during the first 24 h of biodegradation. Four different bacteria, identified as Sphingobacterium spiritivorum, Ochrobactrum anthropi, Bosea thiooxidans and Sphingomonas paucimobilis, were isolated. Pure strains of B. thiooxidans and S. paucimobilis degraded lindane after 3 days of aerobic incubation. This is the first report of lindane biodegradation by B. thiooxidans.     

Eukaryotic diversity in historical soil samples

The eukaryotic biodiversity in historical air-dried samples of Dutch agricultural soil has been assessed by random sequencing of an 18S rRNA gene library and by denaturing gradient gel electrophoresis. Representatives of nearly all taxa of eukaryotic soil microbes could be identified, demonstrating that it is possible to study eukaryotic microbiota in samples from soil archives that have been stored for more than 30 years at room temperature. In a pilot study, 41 sequences were retrieved that could be assigned to fungi and a variety of aerobic and anaerobic protists such as cercozoans, ciliates, xanthophytes (stramenopiles), heteroloboseans, and amoebozoans. A PCR-denaturing gradient gel electrophoresis analysis of samples collected between 1950 and 1975 revealed significant changes in the composition of the eukaryotic microbiota.     

Addition of allochthonous fungi to a historically contaminated soil affects both remediation efficiency and bacterial diversity

Botryosphaeria rhodina DABAC P82 and Pleurotus pulmonarius CBS 664.97 were tested for their ability to grow and to degrade aromatic hydrocarbons in an aged contaminated soil. To evaluate the impact of indigenous microflora on the overall process, incubations were performed on both fumigated and nonfumigated soils. Fungal colonization by B. rhodina was unexpectedly lower in the fumigated than in the nonfumigated soil while the growth of P. pulmonarius showed an opposite response. Degradation performances and detoxification by both fungi in the nonfumigated soil were markedly higher than those observed in the fumigated one. Heterotrophic bacterial counts in nonfumigated soil augmented with either B. rhodina or P. pulmonarius were significantly higher than those of the corresponding incubation control (6.7 ± 0.3 × 10⁸ and 8.35 ± 0.6 × 10⁸, respectively, vs 9.2 ± 0.3 × 10⁷). Bacterial communities of both incubation controls and fungal-augmented soil were compared by numerical analysis of denaturing gradient gel electrophoresis profiles of polymerase chain reaction (PCR)-amplified 16S ribosomal RNA (rRNA) genes and cloning and sequencing of PCR-amplified 16S rRNA genes. Besides increasing overall diversity, fungal augmentation led to considerable qualitative differences with respect to the pristine soil.     

Assessing the bacterial diversity and functional profiles of the River Yamuna using Illumina MiSeq sequencing

Archives of Microbiology - A small percentage of the total freshwater on Earth is represented by river water. Microbes have an essential role to play in the biogeochemical cycles, mineralization of...     

Ultraviolet irradiation of soil samples as a model of the effect of stress factors on bacterial diversity in soil ecosystem

UV irradiation is proposed for use in studying the effect of radioactive irradiation, since radioresistant bacteria are, as a rule, resistant to UV, and the mechanisms of repair of cell damage induced by UV and ionizing radiation are similar. It was found that the total number of bacteria and the number of dominant species in soil samples exposed to UV radiation decreased, indicating the unfavorable effect of UV radiation on bacterial diversity in soil ecosystems. The percentage of cells of bacteria belonging to dominant species varied significantly depending on the intensity of UV irradiation. It can be inferred that long-term irradiation of soils must impair the stability of soil ecosystems, a phenomenon that was indeed observed in the zone around the Chernobyl Nuclear Power Plant. At the same time, the UV irradiation of soil samples made it possible to reveal minor species, primarily UV-resistant pigmented bacteria. UV irradiation can probably be used as a selective factor for the isolation of radioresistant species.     

Antagonistic and antimicrobial activities of some bacterial isolates collected from soil samples

Thirty seven bacterial cultures isolated from soil samples obtained from different locations were tested for their antagonistic activity against some fungal pathogens, viz., Sclerotium rolfsii, Fusarium oxysporum and Rhizoctonia solani, causal agents of collar rot of sunflower, wilts and root rots, respectively. Among them, 5 bacterial strains, viz., A1 6 (Bacillus sphaericus), K1 24 (Pseudomonas fluorescens), M1 42 (Bacillus circulans), M1 66 (Bacillus brevis) and T1 22 (Bacillus brevis) showed positive antagonistic activity. M1 66 was the most effective in inhibiting mycelial growth of S. rolfsii in vitro followed by M1 42, T1 22, K1 24 and A1 6. Only one bacterial strain i.e. M1 42 exhibited antagonistic activity against F. oxysporum, and none of the bacterial strains gave positive activity against R. solani. Furthermore, antimicrobial activities of all the 5 strains were checked against different test organisms. These strains showed their extensive inhibition effect particularly against gram-positive test bacteria (Staphylococcus aureus and Bacillus subtilis) and the test fungal strain (Candida albicans). On the other hand, B. brevis M1 66 and B. brevis T1 22 strains had an inhibitory effect against gram positive and gram-negative test bacteria (Escherichia coli and Proteus vulgaris) as well as the test fungal strain.     

Modeling soil bacterial diversity: challenges and opportunities

Understanding the mechanisms structuring soil bacterial diversity has critical implications to advance the parametrization of species distribution models. This forum article discusses recent advances in the use of the metabolic theory of ecology applicable to soil microbiology, and highlights challenges and opportunities to inform future empirical and theoretical studies.     

砷还原菌群对砷的还原作用及菌群的多样性分析

从砷污染土壤中富集砷抗性细菌,在厌氧环境中进行培养,观察其对砷的还原能力。结果表明：在21 h之内,As(V)就被完全还原为As(Ⅲ);培养72 h后,培养基中出现黄色沉淀,采用X射线衍射分析(XRD)和扫描电镜-能谱分析(SEM-EDS)技术对沉淀进行分析表明,沉淀主要是以3种晶型存在的硫化砷(AsS);培养150 h后,大约有65%的As以上述沉淀的方式从溶液中移除。此外,本文还采用了构建16S rDNA文库的方式对该体系中的微生物种群进行分析,利用RFLP技术对16S rDNA片段进行分型,共得到72个操作单元类型(OTU),其中6个OTU占了库容的51%,从这6个OTU中各选取1个克隆进行测序,结果表明,富集到的砷还原细菌属于喜热菌属(Caloramator)、梭菌属(Clostridium)和杆菌属(Bacillus)。     

Analysis of the bacterial strains using Biolog plates in the contaminated soil from Riyadh community

Routine manufacture, detonation and disposal of explosives in land and groundwater have resulted in complete pollution. Explosives are xenobiotic compounds, being toxic to biological systems, and their recalcitrance leads to persistence in the environment. The methods currently used for the remediation of explosive contaminated sites are expensive and can result in the formation of toxic products. The present study aimed to investigate the bacterial strains using the Biolog plates in the soil from the Riyadh community. The microbial strains were isolated using the spread plate technique and were identified using the Biolog method. In this study we have analyzed from bacterial families of soil samples, obtained from the different sites in 5 regions at Explosive Institute. Our results conclude that Biolog MicroPlates were developed for the rapid identification of bacterial isolates by sole-carbon source utilization and can be used for the identification of bacteria. Out of five communities, only four families of bacteria indicate that the microbial community lacks significant diversity in region one from the Riyadh community in Saudi Arabia. More studies are needed to be carried out in different regions to validate our results.     

一株高效降解芘的细菌分离、鉴定及其降解效果

摘要：【目的】获得高效降解高分子量多环芳烃的细菌,并研究其对多环芳烃的降解能力。【方法】利用富集培养和芘升华平板方法,从焦化厂污染土壤中分离多环芳烃降解细菌,对分离菌株通过形态特征、16S rRNA基因和gyrb基因序列相似性分析进行鉴定,并研究该菌对高分子量多环芳烃（HMW-PAHs）的降解效果。【结果】筛选到一株能以芘、苯并蒽、屈、苯并芘、茚并芘、苯并苝、荧恩为碳源和能源生长并降解这些底物的菌株HBS1,该菌株的16S rRNA基因和gyrb基因序列与Gordonia amicalis的相应基因的相似     

Diversity and PAH growth abilities of bacterial strains isolated from a contaminated soil in Slovakia

Different abandoned industrial areas contaminated by polycyclic aromatic hydrocarbons (PAHs) are present in Slovakia. These environmental burdens are very dangerous to the health of human and environment. The bioremediation, based on the use of hydrocarbons degrading microorganisms, is a promising strategy to sanitize these polluted sites. The aim of this investigation was to assess the bacterial diversity of a PAHs-contaminated soil and to select the potential hydrocarbonoclastic bacteria which can be used for different bioremediation approaches. The bacterial strains were isolated on minimal medium agar supplemented with a mixture of PAHs. Seventy-three isolated strains were grouped by ribosomal interspacer analysis in 15 different clusters and representatives of each cluster were identified by 16S rRNA sequencing. The PAHs degradation abilities of all bacterial isolates were estimated by the 2,6-dichlorophenol indophenol assay and by their growth on minimal broth amended with a mixture of PAHs. Different kinds of strains, members of the genus Pseudomonas, Enterobacter, Bacillus, Arthrobacter, Acinetobacter and Sphingomonas, were isolated from the contaminated soil. Four isolates (Pseudomonas putida, Arthrobacter oxydans, Sphingomonas sp. and S. paucimobilis) showed promising PAHs-degrading abilities and therefore their possible employing in bioremediation strategies.     

马里亚纳海沟共培养细菌多样性动态变化及潜在微生物互作关系

马里亚纳海沟是世界已知最深的海沟,其寡营养、高压、低温、低氧等极端的深海环境孕育出独特的细菌群落结构及多样性特征。选取寡营养培养基对马里亚纳海沟海水及表层沉积物分别进行液体共培养,并在不同培养阶段取样进行高通量测序,分析细菌群落结构组成及其多样性的动态变化,探讨微生物之间可能的互作关系。研究结果表明：液体共培养样品中一共检测到19个门、34个纲、76个目、131个科、227个属的细菌,其中变形菌门（Proteobacteria）和拟杆菌门（Bacteroidetes）为优势菌群,其次为厚壁菌门（Firmicutes）;与其他样品相比,1000米海水样品中细菌群落的多样性最高,并且蓝细菌门（Cyanobacteria）具有更高的相对丰度。共培养样品中细菌丰富度、多样性、群落结构均随培养时间而改变,其中共培养中期样品的细菌多样性较高;表层沉积物样本中,盐单胞菌属（Halomonas）可能由于较强的竞争能力在共培养后期占据优势地位。基因功能预测与代谢通路富集结果显示,随着共培养时间的增加,微生物生长相关的代谢通路丰度明显下降,而与互作相关的代谢通路丰度明显增加。共培养样品检测到的细菌多样性远高于单独分离培养的多样性,仅有少量菌属为单独分离培养与共培养样品均检测到的共有属。综上所述,马里亚纳海沟细菌群落中存在竞争、互利共生的相互作用,共培养法有利于揭示细菌间的互作关系。研究为深渊及深海等极端环境下微生物生态系统组成及维持奠定了理论基础,也为进一步研究极端微生物的生存策略提供了科学指导。     

Bacterial diversity and reductive dehalogenase redundancy in a 1,2-dichloroethane-degrading bacterial consortium enriched from a contaminated aquifer

Background  

Bacteria possess a reservoir of metabolic functionalities ready to be exploited for multiple purposes. The use of microorganisms to clean up xenobiotics from polluted ecosystems (e.g. soil and water) represents an eco-sustainable and powerful alternative to traditional remediation processes. Recent developments in molecular-biology-based techniques have led to rapid and accurate strategies for monitoring and identification of bacteria and catabolic genes involved in the degradation of xenobiotics, key processes to follow up the activities in situ.     

Phenotypic diversity and antibiotic resistance in soil bacterial communities

The community structure in two different agricultural soils has been investigated. Phenotypic diversity was assessed by applying BIOLOG-profiles on a total of 208 bacterial isolates. Diversity indices were calculated from cluster analysis of the BIOLOG data. The bacterial isolates were also evaluated for resistance towards six different antibiotics, mercury resistance and the presence of plasmids. The presence of tetracycline-resistant determinants class A to E among Gram-negative bacteria was analysed with DNA probes. The distribution of tetracycline resistance markers among colonies growing on non-selective and tetracycline-selective plates were compared. The phenotypic approach demonstrated some difference in the diversity within the two soils. The frequency of antibiotic resistance isolates was high in both soils, whereas the frequency of mercury resistance differed significantly. We found no correlation between plasmid profiles and antibiotic resistance patterns. We found all the tetracycline resistance determinants except class B, indicating that the diversity of the tetracycline resistance determinants was complex in populations of resident soil bacteria under no apparent selective pressure for the genes in question.     

紫茎泽兰入侵对土壤细菌的群落组成和多样性的影响

外来生物入侵可能对生物群落结构和生态系统功能产生多种影响, 但入侵植物与土壤微生物群落组成和多样性的关系尚不清楚。为了揭示外来植物紫茎泽兰(Eupatorium adenophorum)入侵对土壤化学性质和细菌群落组成及多样性的影响, 本研究利用第二代高通量测序技术, 比较了紫茎泽兰不同入侵程度的生境(本地植物群落、紫茎泽兰与本地植物混生群落、紫茎泽兰单优群落)土壤中细菌群落的差异。土壤化学性质分析表明, 土壤pH值、有机质、全N和全K随着紫茎泽兰的入侵而逐渐降低, 而土壤全P则在入侵程度最高的生境土壤中最高。通过测序共获得7,755个细菌OUT (operational taxonomic unit)。结果表明, 紫茎泽兰入侵对土壤的细菌多样性影响较小, ACE和Chao指数在3种不同生境间的差异不显著。细菌在紫茎泽兰与本地植物混生群落中的Shannon指数最低, 即细菌的多样性在中等入侵程度的生境最低。此外, 紫茎泽兰入侵改变了土壤细菌组成和结构, 酸杆菌门(Acidobacteria)和疣微菌门(Verrucomicrobia)的相对丰度, 从本地植物群落、混合群落到紫茎泽兰单优群落, 呈现出先增加后减少的趋势。可见, 紫茎泽兰入侵一定程度上改变了土壤微生物的多样性和群落结构, 并改变了土壤的化学性质。     

Characterization of bacterial strains capable of desulphurisation in soil and sediment samples from Antarctica

The presence of sulphur in fossil fuels and the natural environment justifies the study of sulphur-utilising bacterial species and genes involved in the biodesulphurisation process. Technology has been developed based on the natural ability of microorganisms to remove sulphur from polycyclic aromatic hydrocarbon chains. This biotechnology aims to minimise the emission of sulphur oxides into the atmosphere during combustion and prevent the formation of acid rain. In this study, the isolation and characterization of desulphurising microorganisms in rhizosphere and bulk soil samples from Antarctica that were either contaminated with oil or uncontaminated was described. The growth of selected isolates and their capacity to utilise sulphur based on the formation of the terminal product of desulphurisation via the 4S pathway, 2-hydroxybiphenyl, was analysed. DNA was extracted from the isolates and BOX-PCR and DNA sequencing were performed to obtain a genomic diversity profile of cultivable desulphurising bacterial species. Fifty isolates were obtained showing the ability of utilising dibenzothiophene as a substrate and sulphur source for maintenance and growth when plated on selective media. However, only seven genetically diverse isolates tested positive for sulphur removal using the Gibbs assay. DNA sequencing revealed that these isolates were related to the genera Acinetobacter and Pseudomonas.