Bacterial community structure, compartmentalization and activity in a microbial fuel cell

AIMS: To characterize bacterial populations and their activities within a microbial fuel cell (MFC), using cultivation-independent and cultivation approaches. METHODS AND RESULTS: Electron microscopic observations showed that the fuel cell electrode had a microbial biofilm attached to its surface with loosely associated microbial clumps. Bacterial 16S rRNA gene libraries were constructed and analysed from each of four compartments within the fuel cell: the planktonic community; the membrane biofilm; bacterial clumps (BC) and the anode biofilm. Results showed that the bacterial community structure varied significantly between these compartments. It was observed that Gammaproteobacteria phylotypes were present at higher numbers within libraries from the BC and electrode biofilm compared with other parts of the fuel cell. Community structure of the MFC determined by analyses of bacterial 16S rRNA gene libraries and anaerobic cultivation showed excellent agreement with community profiles from denaturing gradient gel electrophoresis (DGGE) analysis. CONCLUSIONS: Members of the family Enterobacteriaceae, such as Klebsiella sp. and Enterobacter sp. and other Gammaproteobacteria with Fe(III)-reducing and electrochemical activity had a significant potential for energy generation in this system. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has shown that electrochemically active bacteria can be enriched using an electrochemical fuel cell.     

A novel three-compartmented electrochemical bioreactor for enrichment of strict anaerobes based on metabolite production

In this study, a novel three-compartmented electrochemical bioreactor (3-CEB) was designed in an effort to overcome the disadvantages of the two-compartmented electrochemical bioreactor (2-CEB) separated with a cation-selective membrane for enrichment of strict anaerobes. The 3-CEB was comprised of an anode, outlet, and a cathode compartment. The outlet compartment was positioned between the anode and cathode compartment, and it was separated with the anode side by a rubber plate and with the cathode side by a porous glass membrane. A platinum wire bridging the anode and outlet compartment operated as a redox passage, however, through which no material could permeate. Butyrate fermentation bacteria were enriched on the basis of the metabolite production. Butyrate generated by strict anaerobes was significantly more abundant in the 3-CEB than in the 2-CEB. Acetic acid and lactic acid generated by facultative anaerobes was relatively higher in the 2-CEB than in the 3-CEB. Meanwhile, butyrate was not generated in the bioreactor utilized for the control test, to which the electrochemical potential was not charged. In a continuous culture using the 3-CEB, the majority of the glucose was fermented to butyrate, and the acetate additionally supplied to the bacterial culture was metabolically reduced to butyrate. More lactate than butyrate was generated from glucose in the 2-CEB.     

Molecular hydrogen from water radiolysis as an energy source for bacterial growth in a basin containing irradiating waste

Although being deionized, filtered and therefore normally deeply oligotrophic, the water from a basin containing irradiating waste presented relatively high bacterial concentrations (ca 10(5) cfu ml(-1)) and biofilm development at its surface and on the walls. This water was characterized by a high concentration of molecular H2 due to water radiolysis, while its electrochemical potential was around +400 mV due the presence of dissolved O2 and active oxygen compounds. This combination of H2 availability and of an oxidant environment is completely original and not described in nature. From surface and wall biofilms, we enumerated the autotrophic populations ( approximately 10(5) bacteria ml(-1)) able to grow in presence of H2 as energy source and CO2 as carbon source, and we isolated the most abundant ones among cultivable bacteria. They efficiently grew on a mineral medium, in the presence of H2, O2 and CO2, the presence of the three gases being indispensable. Two strains were selected and identified using their rrs gene sequence as Ralstonia sp. GGLH002 and Burkholderia sp. GGLH005. In pure culture and using isotope exchange between hydrogen and deuterium, we demonstrated that these strains are able to oxidize hydrogen as energy source, using oxygen as an electron acceptor, and to use carbon dioxide as carbon source. These chemoautotroph hydrogen-oxidizing bacteria probably represent the pioneer bacterial populations in this basin and could be primary producers in the bacterial community.     

Electrochemical and microbiological characterization of paper mill biofilms

Biofilm samples collected from inside and outside the press and former sections of paper machines in a Northwestern Ontario paper mill for a period of 2 years were characterized microbiologically and electrochemically. Bacterial community profiling was done using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and selected bacterial isolates were identified using 16S rDNA analysis. The bacterial community showed the presence of Proteobacteria, Firmicutes, and Actinobacteria. Sphingomonas sp. was found to be the most common bacterial species, which showed the highest production of extracellular polymeric substances. Bacteria isolated from biofilms showed better adhesion properties than those from water samples. Cyclic voltammetry and electrochemical impedance spectroscopy studies showed that bacteria isolated from biofilms and feed water collected from inside the machine were more easily oxidized than those from outside, suggesting the need for a more rigorous biofilm abatement strategy for inside paper machines.     

A microbial consortium isolated from a crude oil sample that uses asphaltenes as a carbon and energy source

A microbial consortium capable of mineralizing asphaltenes was obtained from the Maya crude oil. The enrichment system was built with a glass column reactor containing mineral medium supplied with asphaltenes as energy and carbon source. The consortium growth was evaluated in Casoy agar during 40 weeks. The steady-state phase of the enriched bacterial community was observed after 10 weeks when the culture reach 10(5) to 10(6) CFU ml(-1). The isolates belong to bacterial genus reported for degradation of other hydrocarbons and they were identified as Corynebacterium sp., Bacillus sp., Brevibacillus sp. and Staphylococcus sp. The bacterial consortium growth was evaluated by a viable counts during 14 days exposed to different aeration, temperature, salinity, and pH conditions. The ability of the consortium to mineralize asphaltenes was evaluated using the method of ISO 9439 in glass column reactors of 20 x 3.2 cm during 13 days. Temperatures of 55 degrees C and salinity of 1.8% were growth limiting. The respiration of the microbial consortium using asphaltenes as a sole carbon source (800 micromoles CO2 in 13 days) was significantly higher than those of the samples containing only the microbial consortium (200 micromoles CO2) or only asphaltenes (300 micromoles CO2). These results indicated the existence of asphaltenes-degradating microbes in the crude oil and confirmed that the consortium could mineralize asphaltenes in conditions of room temperature, salinity of 100 ppm, aeration of 1 l min(-1) and pH of 7.4.     

Microbial utilization of electrically reduced neutral red as the sole electron donor for growth and metabolite production.

Electrically reduced neutral red (NR) served as the sole source of reducing power for growth and metabolism of pure and mixed cultures of H2-consuming bacteria in a novel electrochemical bioreactor system. NR was continuously reduced by the cathodic potential (-1.5 V) generated from an electric current (0.3 to 1.0 mA), and it was subsequently oxidized by Actinobacillus succinogenes or by mixed methanogenic cultures. The A. succinogenes mutant strain FZ-6 did not grow on fumarate alone unless electrically reduced NR or hydrogen was present as the electron donor for succinate production. The mutant strain, unlike the wild type, lacked pyruvate formate lyase and formate dehydrogenase. Electrically reduced NR also replaced hydrogen as the sole electron donor source for growth and production of methane from CO2. These results show that both pure and mixed cultures can function as electrochemical devices when electrically generated reducing power can be used to drive metabolism. The potential utility of utilizing electrical reducing power in enhancing industrial fermentations or biotransformation processes is discussed.     

Temperature and biotic factors influence bacterial communities associated with the cyanobacterium Microcystis sp

Cyanobacterial blooms represent a nutritious niche for associated bacteria including potential pathogens for humans as well as livestock. We investigated bacterial community composition associated with Microcystis sp. using different approaches: batch experiments on Microcystis sp. or its enriched exudates, field enclosures (dialysis bags) and field sampling during natural blooms in freshwaters. Bacterial community composition associated with Microcystis sp. differed significantly with temperature, bacterial source community and number of incubated cyanobacterial strains. Interestingly, Actinobacteria of the AcI cluster were only present in the 20°C treatments and disappeared at higher incubation temperatures. Moreover, Archaea were present in all field samples but did not show any regional patterns, which is consistent with bacteria. Absence of Archaea in the experimental treatments indicates reduced growth under experimental conditions. In contrast, members of the genus Sphingomonas (Alphaproteobacteria), which includes species known as human pathogens, occurred in almost all samples. Thus Sphingomonadales seem to be an integral element of Microcystis sp. blooms - even affecting concentrations of microcystins as a result of their breakdown of the toxins. Depending on environmental conditions such as temperature, light, currents and nutrients, the role of heterotrophic Bacteria associated with Cyanobacteria can greatly vary by either increasing (pathogens) or decreasing (breakdown of toxins) health risks caused by mass developments of potentially toxic Cyanobacteria.     

Microbial structure and community of RBC biofilm removing nitrate and phosphorus from domestic wastewater

Using a rotating biological contactor modified with a sequencing bath reactor system (SBRBC) designed and operated to remove phosphate and nitrogen [58], the microbial community structure of the biofilm from the SBRBC system was characterized based on the extracellular polymeric substance (EPS) constituents, electron microscopy, and molecular techniques. Protein and carbohydrate were identified as the major EPS constituents at three different biofilm thicknesses, where the amount of EPS and bacterial cell number were highest in the initial thickness of 0-100 mum. However, the percent of carbohydrate in the total amount of EPS decreased by about 11.23%, whereas the percent of protein increased by about 11.15% as the biofilm grew. Thus, an abundant quantity of EPS and cell mass, as well as a specific quality of EPS were apparently needed to attach to the substratum in the first step of the biofilm growth. A FISH analysis revealed that the dominant phylogenetic group was beta- and gamma-Proteobacteria, where a significant subclass of Proteobacteria for removing phosphate and/or nitrate was found within a biofilm thickness of 0-250 mum. In addition, 16S rDNA clone libraries revealed that Klebsiella sp. and Citrobacter sp. were most dominant within the initial biofilm thickness of 0-250 mum, whereas sulfur-oxidizing bacteria, such as Beggiatoa sp. and Thiothrix sp., were detected in a biofilm thickness over 250 mum. The results of the bacterial community structure analysis using molecular techniques agreed with the results of the morphological structure based on scanning electron microscopy. Therefore, the overall results indicated that coliform bacteria participated in the nitrate and phosphorus removal when using the SBRBC system. Moreover, the structure of the biofilm was also found to be related to the EPS constituents, as well as the nitrogen and phosphate removal efficiency. Consequently, since this is the first identification of the bacterial community and structure of the biofilm from an RBC simultaneously removing nitrogen and phosphate from domestic wastewater, and it is hoped that the present results may provide a foundation for understanding nitrate and phosphate.     

Different Marine Heterotrophic Nanoflagellates Affect Differentially the Composition of Enriched Bacterial Communities

We studied the effects of predation on the cytometric and phylogenetic features of two enriched bacterial communities obtained from two cultures of marine heterotrophic nanoflagellates: Jakoba libera and a mixed culture of Cafeteria sp. and Monosiga sp. Protists were harvested by flow cytometric cell sorting and eight different treatments were prepared. Each bacterial community was incubated with and without protists, and we added two treatments with protists and the bacteria present after the sorting procedure (cosorted bacteria). The bacterial community derived from the culture of Jakoba libera had higher green fluorescence per cell (FL1) than that derived from the mixed culture of Cafeteria sp. and Monosiga sp. When the experiment began all treatments presented bacterial communities that increase in fluorescence per bacterium (FL1); after that the FL1 decreased when bacteria attained maximal concentrations; and, finally, there was a new increase in FL1 toward the end of the experiment. Cosorted bacteria of Jakoba libera had the same fluorescence as the bacterial community derived from this protist, while the bacteria derived from the mixed culture of Cafeteria sp. and Monosiga sp. was nearly twice as fluorescent than that of the parental community. All treatments presented a general decline of SSC along the incubation. Therefore, there was a small influence of protists on the cytometric signature of each bacterial community. However, each bacterial community preyed by Jakoba libera or the mixed culture of Cafeteria sp. and Monosiga sp. led to four different phylogenetic fingerprint. Besides, the final Communities were different from the fingerprint of controls without protists, and most of them diverge from the fingerprint of cosorted bacteria. Our results confirm that changes in the phylogenetic composition of marine bacterial communities may depend on the initial communities of both bacteria and protists.     

Elucidating the key member of a linuron-mineralizing bacterial community by PCR and reverse transcription-PCR denaturing gradient gel electrophoresis 16S rRNA gene fingerprinting and cultivation

A bacterial community from Danish agricultural soil was enriched with linuron [N-(3,4-dichlorophenyl)-N'-methoxy-N'-methylurea] as the sole carbon and nitrogen source. The community mineralized [ring-U-14C]linuron completely to 14CO2 and 14C-biomass. Denaturing gradient gel electrophoresis analysis and cultivation revealed that a Variovorax sp. was responsible for the mineralization activity.     

Removal of chromium and pentachlorophenol from tannery effluents

Three bacterial strains, including one Acinetobacter sp. PCP3, grown in the presence of minimal salt medium and pentachlorophenol (PCP) as sole carbon source in the chemostat showed higher utilization of PCP and adsorption of chromium. In sequential bioreactor, tannery effluents treated initially by bacterial consortium followed by fungus removed 90% and 67% chromium and PCP respectively, whereas in another set of bioreactor in which effluents was treated initially by fungi followed by bacteria could remove 64.7% and 58% chromium and PCP, respectively.     

Ectomycorrhizal exudates and pre-exposure to elevated CO2 affects soil bacterial growth and community structure

Ectomycorrhizal fungi produce low molecular weight organic compounds, supporting diverse microbial communities. To link mycorrhizal root exudation directly to bacterial responses, we used Scots pine exudates with (Suillus variegatus and Piloderma fallax) and without mycorrhiza as substrata for forest soil bacteria. Bacterial growth and vitality was monitored, and community composition determined using T-RFLP, cloning and sequencing. We investigated if the amount of organic acids in exudates explained bacterial growth, and whether bacterial communities were influenced by pre-exposure to elevated atmospheric CO₂. We demonstrated functional differences in bacterial growth rates related to CO₂. There was a shift in the bacterial community (e.g. Burkholderia sp. and gamma-proteobacteria) toward organisms better able to rapidly utilize exudates when pine microcosms were pre-exposed to elevated CO₂. Soil bacteria from all treatments tended to grow more abundantly and rapidly in exudates from Piloderma-colonized seedlings, suggesting that the organic acids and/or unidentified compounds present supported greater growth.     

Compositions of microbial communities in sulfide nickel ore waste piles

Bacterial communities of moderately acidic waste piles of sulfide nickel ore and of the nickel-leaching enrichments obtained from them are analyzed. The structure of bacterial communities was determined by molecular biological techniques. The PCR profiles of bacterial communities were obtained with the primers to a variable 433 bp site of the eubacterial 16S rRNA gene. The differences in community compositions were determined by comparison of their DGGE profiles. Sequencing of the DNA fragments was then carried out and the results were compared with the GenBank gene sequences. Analysis of the 16S rRNA gene sequences revealed few bacterial genera in the moderately acidic waste piles of sulfide nickel ore, with predomination of Acidithiobacillus sp. and Leptospirillum sp. A number of the bacteria revealed belonged to the species never obtained in pure culture. Molecular biological analysis showed the presence of the same groups of bacteria in enriched cultures obtained by inoculating the liquid medium containing ground ore with the waste pile samples (5 : 1). The geochemical activity of these bacteria was confirmed by their capacity for leaching nickel from the sulfide ore in enriched cultures, resulting in nickel solubilization. Thus, new information was obtained concerning the structure composition of the bacterial communities of sulfide ore waste piles: the dominant forms were determined, their leaching activity was confirmed, and the activity of thiobacilli from the waste, which have not been isolated in pure cultures, was confirmed in liquid medium in the presence of ore.     

Dynamics of bacterial community in up-flow anaerobic packed bed system for acid mine drainage treatment using wine wastes as carbon source

The dynamics of the bacterial populations in an up-flow anaerobic packed bed system (UAPB), applied in acid mine drainage treatment using wine wastes as carbon and nutrients source was elucidated by temperature gradient gel electrophoresis (TGGE) analysis. Moreover, TGGE fingerprints of the bacterial communities developed in a UAPB fed with wine wastes and a UAPB fed with pure ethanol were compared. TGGE fingerprinting and phylogenetic analysis showed that the composition of the community in the UAPB fed with wine wastes remained stable during whole time of operation and its bacterial diversity was higher. The bacterial community of the UAPB fed with wine wastes was composed by bacteria affiliated with Desulfovibrio, Clostridium, Citrobacter and Cronobacter genera and with Bacteroidales order, sp. The dominant community developed in the UAPB fed with ethanol was composed by bacteria affiliated with Desulfovibrio sp. The presence of several bacterial groups in the bioreactor fed with wine wastes suggests a synergistic interaction between the different populations. Syntrophic interaction may be the key factor for the utilization of wine wastes, a complex organic substrate, as carbon and electron source for sulphate reduction.     

Effects of ocean acidification on microbial community composition of, and oxygen fluxes through, biofilms from the Great Barrier Reef

Rising anthropogenic CO(2) emissions acidify the oceans, and cause changes to seawater carbon chemistry. Bacterial biofilm communities reflect environmental disturbances and may rapidly respond to ocean acidification. This study investigates community composition and activity responses to experimental ocean acidification in biofilms from the Australian Great Barrier Reef. Natural biofilms grown on glass slides were exposed for 11 d to four controlled pCO(2) concentrations representing the following scenarios: A) pre-industrial (～300 ppm), B) present-day (～400 ppm), C) mid century (～560 ppm) and D) late century (～1140 ppm). Terminal restriction fragment length polymorphism and clone library analyses of 16S rRNA genes revealed CO(2) -correlated bacterial community shifts between treatments A, B and D. Observed bacterial community shifts were driven by decreases in the relative abundance of Alphaproteobacteria and increases of Flavobacteriales (Bacteroidetes) at increased CO(2) concentrations, indicating pH sensitivity of specific bacterial groups. Elevated pCO(2) (C + D) shifted biofilm algal communities and significantly increased C and N contents, yet O(2) fluxes, measured using in light and dark incubations, remained unchanged. Our findings suggest that bacterial biofilm communities rapidly adapt and reorganize in response to high pCO(2) to maintain activity such as oxygen production.     

Development of bacterial community during spontaneous succession on spoil heaps after brown coal mining

Changes in the abundance of bacteria and fungi and in the composition of bacterial communities during primary succession were investigated in a brown coal mine deposit area near Sokolov, the Czech Republic, using phospholipid fatty acids analysis, microarray and 16S rRNA gene sequencing. The study considered a chronosequence of sites undergoing spontaneous succession: 6-, 12-, 21- and 45-year-old and a 21-year-old site revegetated with Alnus glutinosa. During succession, organic carbon and the total nitrogen content increased while the pH and the C/N ratio decreased. Microbial biomass and bacterial diversity increased until 21 years and decreased later; bacteria dominated over fungi in the initial and late phases of succession. Bacterial community composition of the 6-year-old site with no vegetation cover largely differed from the older sites, especially by a higher content of Gammaproteobacteria, Cyanobacteria and some Alphaproteobacteria. Bacteria belonging to the genera Acidithiobacillus, Thiobacillus and related taxa, the CO(2) and N(2) fixers, dominated the community at this site. In the later phases, bacterial community development seemed to reflect more the changes in soil nutrient content and pH than vegetation with a decrease of Actinobacteria and an increase of Acidobacteria. The site revegetated with A. glutinosa resembled the 45-year-old primary succession site and exhibited an even lower pH and C/N ratio, indicating that recultivation is able to accelerate soil development.     

Power generation from cellulose using mixed and pure cultures of cellulose-degrading bacteria in a microbial fuel cell

Microbial fuel cells (MFCs) have been used to generate electricity from various organic compounds such as acetate, glucose, and lactate. We demonstrate here that electricity can be produced in an MFC using cellulose as the electron donor source. Tests were conducted using two-chambered MFCs, the anode medium was inoculated with mixed or pure culture of cellulose-degrading bacteria Nocardiopsis sp. KNU (S strain) or Streptomyces enissocaesilis KNU (K strain), and the catholyte in the cathode compartment was 50mM ferricyanide as catholyte. The power density for the mixed culture was 0.188mW (188mW/m(2)) at a current of 0.5mA when 1g/L cellulose was used. However, the power density decreased as the cellulose concentration in the anode compartment decreased. The columbic efficiencies (CEs) ranged from 41.5 to 33.4%, corresponding to an initial cellulose concentration of 0.1-1.0g/L. For the pure culture, cellobioase enzyme was added to increase the conversion of cellulose to simple sugars, since electricity production is very low. The power densities for S and K strain pure cultures with cellobioase were 162mW/m(2) and 145mW/m(2), respectively. Cyclic voltammetry (CV) experiments showed the presence of peaks at 380, 500, and 720mV vs. Ag/AgCl for the mixed bacterial culture, indicating its electrochemical activity without an external mediator. Furthermore, this MFC system employs a unique microbial ecology in which both the electron donor (cellulose) and the electron acceptor (carbon paper) are insoluble.     

微生物电解池阳极生物膜功能菌群构建及群落特征分析

【目的】微生物电解电池(MEC)是近几年快速发展的利用电极呼吸微生物快速降解有机质,通过较小的辅助外加电压直接生成氢气的新工艺。MEC能够有效地富集高效率电子传递功能菌群,是未来工艺放大和快速启动的关键。【方法】采用不同驯化方法构建MEC电极微生物菌群,通过单链构象多肽性技术(Single-strand conformation poly-morphism,SSCP)快速检测分析启动后电子传递功能菌群特征。【结果】阳极生物膜接种MEC可以实现2 d的快速启动,库仑效率达到20%以上,7 d获得稳定产氢,氢气转化率达到30%,能量回收效率达到90%以上。通过SSCP群落分析发现,采用微生物燃料电池阳极生物膜构建的MEC主要电子传递功能相关的菌群包括Pseudomonas sp.、Flavobacterium sp.、Ochrobactrum sp.,而直接由产氢MEC阳极生物膜新启动的MEC功能菌群组成丰度更大,包括电子传递效能更高的Desulfovibrio、Pseudomonas和Shewanella成为主要优势电子传递菌群。通过稳定产氢运行,MEC阳极生物膜优势菌群中存在的较大比例的厌氧菌与电子传递辅助菌对体系的快速稳定运行十分重要。【结论】与MFC阳极生物膜相比,MEC生物膜作为启动菌源能够获得多样性更丰富的电极功能菌群,其库仑效率和产氢效率更具优势。     

生姜根系不同生态位细菌群落多样性特征、组成及结构差异

生姜作为常见的调味品和传统中药材,是我国重要的经济作物之一。作为取食部分的生姜块茎与根系直接相连,其产量、品质与根相关细菌群落密切相关。然而,关于生姜根系微环境中细菌群落的特点仍鲜有报道,土壤环境能否衍生出宿主特异性内生菌群落尚不清楚。以生姜根系不同生态位细菌群落为研究对象,采用高通量测序技术,对非根际、根际及根内细菌进行16S rRNA基因测序。结果表明,不同生态位细菌群落多样性存在显著差异,其中非根际及根际细菌群落多样性(Shannon index, Observed species, Faith′s PD)显著高于内生菌群落。同时,各生态位共现网络稳定性和复杂度表现为非根际>根际>根内细菌群落。而在组成上,细菌群落在不同生态位差异显著(R²=0.57,P=0.001)。其中变形菌门(Proteobacteria)是根内的优势门,该门类下假单胞菌属(Pseudomonas)、短波单胞菌属(Brevundimonas)、寡养单胞菌属(Stenotrophomonas)及泛菌属(Pantoea)在根内显著富集。在根际细菌中,拟杆菌门(Bacteroid...     

Characterization of alkalotolerant bacterial community by 16S rDNA-based denaturing gradient gel electrophoresis method for degradation of dibenzofuran in soil microcosm

An alkalotolerant bacterial community was developed by continuous enrichment in the chemostat in presence of dibenzofuran (DF) as sole carbon source. Six different types of bacterial isolates were cultured on nutrient broth agar plates together with six operational taxonomic units (OTUs) at pH 7.0 and pH 8.0 by 16S rDNA-DGGE method. However, isolates of microbial community was declined from three OTUs (pH 9.0) to two at pH 10.0 after enrichment in alkaline condition. Among the six isolates tested for degradation of DF, Pseudomonas sp. and Bacillus sp. the members of alkalotolerant bacterial community had better potency to degrade dibenzofuran. Alkalotolerant bacterial community introduced in soil microcosm for evaluation of survival of most suitable isolates and degradation of dioxin-like compound indicated more than 90% degradation of dibenzofuran after 45 days by the bacterial community enriched for 180 days in the chemostat at pH 10, however, microbial community was not competent to utilize even 50% DF after day 30, not enriched in the chemostat. The survival of competent bacteria monitored by DGGE method in soil microcosm indicated presence of two major alkalotolerant isolates for utilization of dibenzofuran, substantiated the results and significance of alkalotolerant bacteria for in situ bioremediation of dioxin-like compounds in the environment.