Biodegradation of the major color containing compounds in distillery wastewater by an aerobic bacterial culture and characterization of their metabolites

This study deals the biodegradation of the major color containing compounds extracted from distillery wastewater (DWW) by an aerobic bacterial consortium comprising Bacillus licheniformis (DQ79010), Bacillus sp. (DQ779011) and Alcaligenes sp. (DQ779012) and characterization of metabolic products. The degradation of color containing compounds by bacteria was studied by using the different carbon and nitrogen sources at different environmental conditions. Results revealed that the bacterial consortium was efficient for 70% color removal in presence of glucose (1.0%) and peptone (0.1%) at pH 7.0 and temperature 37°C. The HPLC analysis of control and bacterial degraded samples has shown the reduction in peak area as well as shifting of peaks compared to control indicating the bacterial degradation as well as transformation of color containing compounds from DWW. The comparative LC–MS–MS and other spectrophotometric analysis has shown the presence of dihydroxyconiferyl alcohol, 2, 2′-bifuran-5-carboxylic acid, 2-nitroacetophenone, p-chloroanisol, 2, 3-dimethyl-pyrazine, 2-methylhexane, methylbenzene, 2, 3-dihydro-5-methylfuran, 3-pyrroline, and acetic acid in control samples that were biodegraded and biotransformed into 2-nitroacetophenone, p-chloroanisol, 2, 2′-bifuran, indole, 2-methylhexane, and 2, 3-dihydro-5-methylfuran by bacterial consortium. In this study, it was observed that most of the compounds detected in control samples were diminished from the bacterial degraded samples and compounds 2, 2′-bifuran and indole with molecular weight 134 and 117 were produced as new metabolites during the bacterial degradation of color containing compounds from DWW.     

Phylogenetic characterization of the blue filamentous bacterial community from an Icelandic geothermal spring

Molecular phylogenetic analysis of a blue filamentous community from an alkaline thermal spring (79-83 degrees C) in Iceland revealed that the blue filaments were affiliated with the Aquificales. The dominant sequence type, pIce1, was most closely related to a sequence (SRI-48) found in a white filamentous community from a separate Icelandic thermal spring and the pink filaments (EM17) from Yellowstone National Park. Fluorescent in situ hybridization with clone-specific oligonucleotide probes showed that the sample analyzed was essentially a monoculture of a single phylotype.     

内蒙古退化荒漠草原土壤细菌群落结构特征

用PCR-DGGE技术对不同退化程度荒漠草地土壤细菌群落结构的分析表明:不同退化草地土壤细菌群落结构发生了明显的变化。2006年夏季,不同退化草地土壤细菌的Shannon-Weaver指数随退化程度的加剧呈降低趋势,由高到低依次是:轻度退化、中度退化和重度退化草地。2007年夏季多样性指数随退化程度的加剧呈先降低后增加的趋势,由高到低依次为:轻度退化、重度退化和中度退化。对17个克隆进行了序列测定所得到的16SrDNA在数据库中的登录号为:EU327142-EU327157和EU327164。将其与Genbank数据库中的序列进行比对,结果表明这些序列与已知序列的相似性均在95%-99%之间。系统发育分析结果表明,内蒙古荒漠草地土壤主要细菌隶属于以下几个分支:拟杆菌纲(Bacteroidetes),酸杆菌纲(Acidobacteria),变形菌纲(Proteobacteria)的γ、δ类群和厚壁菌纲(Firmicutes)。其中,拟杆菌纲所占的比例为47%左右,为优势种。     

Solubilization of Leonardite by an Extracellular Fraction from Coriolus versicolor

Coriolus versicolor has previously been shown to degrade leonardite, an oxidized form of lignite. An extracellular fraction containing protein purified from a C. versicolor culture solubilized leonardite in vitro. Expression of the activity did not require the presence of leonardite and appeared during idiophase. During ion-exchange and gel filtration column chromatography, leonardite-biosolubilizing activity eluted with syringaldazine oxidase activity and with protein, as measured by A₂₈₀ and the biuret protein assay. Syringaldazine is a substrate of the polyphenol oxidase formed by C. versicolor. Comparison of leonardite-biosolubilizing activity with the effects of chelators and surface-active agents on leonardite showed that biosolubilization was not due to either surfactant or chelating ability. Heat treatment of the preparation at 60°C for 30 min significantly reduced both syringaldazine oxidase and leonardite-biosolubilizing activities. Cyanide, azide, and thioglycolate, which are known inhibitors of syringaldazine oxidase activity of C. versicolor, also inhibited leonardite biosolubilization. From these data, we conclude that the purified protein fraction from C. versicolor contains a syringaldazine oxidase activity that participates in leonardite biosolubilization by enzymatic action.     

Biodegradation of bisphenol A by bacterial consortia

The effect of light on BPA degradation by an adapted bacterial consortium was investigated. BPA was completely degraded up to 50 mg l⁻¹, and the degradation followed first-order reaction kinetics both in the light and in the dark. The degradation half-life of BPA when the consortium was grown in presence of light was 21.9, 17.2, and 12.6 h for concentrations of 10, 20, and 50 mg l⁻¹, respectively; the degradation half-life of BPA in the dark was 13.1, 10.8, and 10.2 h for concentrations of 10, 20, and 50 mg l⁻¹, respectively. Therefore, light inhibited BPA biodegradation. However, under both conditions, BPA was completely depleted. The bacterial consortium effectively utilised BPA as a growth substrate to sustain a cell yield of 0.95 g g⁻¹ and 0.97 g g⁻¹ in the light and dark, respectively. A total of ten and nine biodegradation intermediates were detected in the light and dark, respectively. Three bacterial metabolic pathways and one photodegradation pathway were proposed to explain their occurrence. This study demonstrated that bacterial consortia may assemble a wide range of catabolic pathways to allow for efficient degradation of BPA, converting BPA to principally bacterial biomass and metabolites exhibiting low or no oestrogenic activity.     

Biodegradation of olive washing wastewater pollutants by highly efficient phenol-degrading strains selected from adapted bacterial community

The bacterial community of an olive washing water (OWW) storage basin was characterized, by both cultivation and cultivation-independent methods. PCR-TGGE fingerprints analysis of different samples, taken along the olive harvesting season, revealed important variations of the bacterial community structure showing rapid establishment of prevalent bacterial populations. Several bacteria, isolated from OWW, were cultivated, in media containing increasing amounts of polyphenols, in order to select high phenol-degrading strains for the effluent pollutants reduction. Strains PM3 and PM15, affiliated to Raoultella terrigena and Pantoea agglomerans by 16S rRNA gene sequencing, were selected and used for OWW biological treatment under batch conditions in shake flasks cultures. The OWW content of phenols, BOD₅, COD and colour, was reduced by 93, 91, 89 and 62%, respectively, permitting effluent disposal and/or reuse with no additional treatments.     

Detoxification of 2,4,6-trichlorophenol by an indigenous bacterial community

An indigenous bacterial community capable of degrading 2,4,6-trichlorophenol (TCP) as the sole carbon source was isolated from the Riachuelo, a polluted river in Buenos Aires. The community consists of three gram-negative bacterial, non-fermentative strains, two of them was identified as belonging to genus Pseudomonas and the other to genus Stenotrophomonas. None of the individual strains were capable of degrading TCP as the sole carbon source. Aerobic biodegradation assays were performed using a 2-l microfermentor at 28 °C with agitation. Biodegradation was evaluated by spectrophotometry, chloride release, gas chromatography and microbial growth. Detoxification was evaluated by using Vibrio fischeri, Pseudokirchneriella subcapitata and Daphnia magna as test organisms. The indigenous bacterial community degrades 100 mg l^?1 of TCP in 27 h. The absence of metabolites and toxicity was proved at the end of the process. The influence of the initial concentration of compound, pH, cell inoculum, presence of other substrates and toxic related compounds in the biodegradation process was assayed. Also the application for polluted water was studied. The promising behavior of the bacterial community under the different conditions assayed allows us to suggest its possible use in remediation processes.     

喀斯特原生土壤与退化生态系统土壤细菌群落结构

运用PCR-RFLP技术,对桂西北喀斯特原生土壤和退化生态系统土壤细菌16S rDNA基因多样性及系统发育关系进行了研究.结果表明：原生土壤比退化生态系统土壤具有更丰富的16S rDNA基因型和更高的多样性指数,两样地共有的基因型仅有2个.从每种基因型中随机选择一个克隆子作为代表进行测序分析,所有序列与GenBank 数据库中序列的同源性为87%～100%,且两样地中均有超过一半的基因型序列与数据库中已知序列同源性低于97%,属于分类在“种”地位上的新发现细菌;通过系统发育研究将两样地的细菌分为10大类群,两样地共同拥有5大类群,但两样地的细菌优势类群明显不同,原生土壤为Proteobacteria,含39种基因型,占总克隆子数的58.0%,退化生态系统土壤为Acidobacteria和Proteobacteria,分别含19种和15种基因型,占总克隆子数的32.5%和30.5%;与原生土壤细菌类群相比,退化生态系统土壤Proteobacteria类群明显减少,Acidobacteria类群明显增加.土壤理化性质及土壤环境因素的差异是引起两类型土壤细菌多样性差异的原因.     

Biodegradation of gasoline by gellan gum-encapsulated bacterial cells

Encapsulated cell bioaugmentation is a novel alternative solution to in situ bioremediation of contaminated aquifers. This study was conducted to evaluate the feasibility of such a remediation strategy based on the performance of encapsulated cells in the biodegradation of gasoline, a major groundwater contaminant. An enriched bacterial consortium, isolated from a gasoline-polluted site, was encapsulated in gellan gum microbeads (16-53 microm diameter). The capacity of the encapsulated cells to degrade gasoline under aerobic conditions was evaluated in comparison with free (non-encapsulated) cells. Encapsulated cells (2.6 mg(cells) x g(-1) bead) degraded over 90% gasoline hydrocarbons (initial concentration 50-600 mg x L(-1)) within 5-10 days at 10 degrees C. Equivalent levels of free cells removed comparable amounts of gasoline (initial concentration 50-400 mg x L(-1)) within the same period but required up to 30 days to degrade the highest level of gasoline tested (600 mg x L(-1)). Free cells exhibited a lag phase in biodegradation, which increased from 1 to 5 days with an increase in gasoline concentration (200-600 x mg L(-1)). Encapsulation provided cells with a protective barrier against toxic hydrocarbons, eliminating the adaptation period required by free cells. The reduction of encapsulated cell mass loading from 2.6 to 1.0 mg(cells) x g(-1) bead caused a substantial decrease in the extent of biodegradation within a 30-day incubation period. Encapsulated cells dispersed within the porous soil matrix of saturated soil microcosms demonstrated a reduced performance in the removal of gasoline (initial concentrations of 400 and 600 mg x L(-1)), removing 30-50% gasoline hydrocarbons compared to 40-60% by free cells within 21 days of incubation. The results of this study suggest that gellan gum-encapsulated bacterial cells have the potential to be used for biodegradation of gasoline hydrocarbons in aqueous systems.     

Biodegradation of hydrocarbon contamination by immobilized bacterial cells

This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.     

一株霍氏肠杆菌对四环素的降解作用及其降解产物的毒性评估

【目的】抗生素作为新兴污染物,已经引起社会的极大关注。针对四环素有效降解菌株缺乏这一现状,本研究旨在筛选和鉴定具有降解四环素功能的菌株,分析其降解特性和降解作用类型、初步探讨其降解活性物质定位并评估其降解产物的生理毒性。【方法】以四环素为唯一碳源,从受四环素污染的猪场污泥中筛选四环素降解菌株;结合菌落形态学特征、生理生化特征、扫描电镜观察和16S rRNA基因测序鉴定菌株,通过不同外源碳、pH及去除动力学阐明菌株对四环素的降解特性,提取菌株不同成分探讨其去除四环素的作用类型,并进一步从细胞内液和细胞外液开展生物降解的活性物质定位,最后评估降解产物的生理毒性。【结果】筛选鉴定得到一株霍氏肠杆菌(Enterobacter hormaechei) MEH2305,pH为7.0和添加10 g/L外源碳胰蛋白胨是其发挥降解作用的最适条件。MEH2305通过非生物降解和生物降解的共同作用,在培养第7天对四环素总去除率达到68% (对土霉素和盐酸强力霉素去除率分别为53%和56%),其分泌的细胞内液和细胞外液对四环素的去除率分别为40.77%和31.18%。同时,与未经MEH2305处理的四环素对照组比较,MEH2305降解四环素的产物对革兰氏阴性大肠杆菌(Escherichia coli) K88和革兰氏阳性枯草芽孢杆菌(Bacillus subtilis) 168的生理毒性作用显著降低。【结论】MEH2305可以作为一株潜在的有效且安全的四环素降解菌株,应用于抗生素的环境治理领域。     

Isolation and characterization of soybean waste-degrading microorganisms and analysis of fertilizer effects of the degraded products.

Two microorganisms which could degrade soybean lees efficiently were isolated and identified as Bacillus circulans and B. stearothermophilus. These two strains secreted thermostable proteases into the medium and could digest soybean lees rapidly and completely at 50 degrees C. Initially, the soybean lees were degraded to proteins in approximately 20 h by these two strains, after which time the concentrations of peptides in the medium gradually increased. The degraded products from soybean lees contained abundant nitrogen compounds, such as peptides, amino acids, and amides. Approximately 10 times more fresh plant weight was obtained (in the case of Brassica campestris) when these degraded products were applied than when water was applied for 42 days. These stimulatory effects of the soybean lees products were almost equal to those of a chemically synthesized fertilizer.     

Biodegradation of methyl t-butyl ether by pure bacterial cultures

Three pure bacterial cultures degrading methyl t-butyl ether (MTBE) were isolated from activated sludge and fruit of the Gingko tree. They have been classified as belonging to the genuses Methylobacterium, Rhodococcus, and Arthrobacter. These cultures degraded 60 ppm MTBE in 1–2 weeks of incubation at 23–25 °C. The growth of the isolates on MTBE as sole carbon source is very slow compared with growth on nutrient-rich medium. Uniformly-labeled [¹⁴C]MTBE was used to determine ¹⁴CO₂ evolution. Within 7 days of incubation, about 8% of the initial radioactivity was evolved as ¹⁴CO₂. These strains also grow on t-butanol, butyl formate, isopropanol, acetone and pyruvate as carbon sources. The presence of these compounds in combination with MTBE decreased the degradation of MTBE. The cultures pregrown on pyruvate resulted in a reduction in ¹⁴CO₂ evolution from [¹⁴C]MTBE. The availability of pure cultures will allow the determination of the pathway intermediates and the rate-limiting steps in the degradation of MTBE. Received: 8 December 1995 / Received last revision: 5 August 1996 / Accepted: 12 August 1996     

Biodegradation of chrysene by the bacterial strains isolated from oily sludge

The biodegradation studies were conducted to test the ability of the bacterial strains (Chry2 and Chry3) isolated from the oily sludge obtained from Gujarat refinery, India, for utilization of chrysene in the liquid medium. Biodegradation of the compound was confirmed using gas chromatography and the percent degradation was calculated to be 15.0 and 17% by Chry2 and Chry3, respectively. The biodegradation results were supported by increase in viable cell count and dry biomass, in the presence of chrysene as the sole carbon source. Both the cultures produced biosurfactant which was indicated by the reduction in surface tension of the growth medium. Presence of catechol 2, 3-dioxygenase gene in Chry3 indicated its potential for degradation of PAHs through meta cleavage degradation pathway. Both the strains were found to possess catechol 1,2-dioxygenase and catechol 2,3-dioxygenase enzyme activities. Based on morphological and biochemical tests, the cultures were tentatively identified as Bacillus sp. (Chry2) and Pseudomonas sp. (Chry3).     

Isolation and characterization of a bluegill-degrading microorganism, and analysis of the root hair-promoting effect of the degraded products

Bluegill-degrading bacteria were isolated from various environmental sources. Brevibacillus sp. BGM1 degraded bluegill efficiently at 50 degrees C, and its culture supernatant showed the highest peptide and amino acid concentrations as trichloroacetic acid (TCA) soluble fraction (ASF) (10.7 mg/ml) of all supernatants obtained with bluegill as a substrate. Strain BGM1 secreted a protease(s) into the medium, and the concentration of peptides and amino acids gradually increased. The fertile effect of the degraded bluegill products (DGP) on Brassica rapa was also investigated. The root hair density of B. rapa grown with DGP at a concentration of 30 mug peptides and amino acids/ml was about 1.7 times higher than when grown with the same concentration of undegraded bluegill. DGP was shown to increase root hair numbers and adventitious root formation. The results of this study suggest that a specific peptide(s) for promotion of root hair is produced from the order Perciformes with a protease(s) from BGM1.     

Biodegradation of a biocide (Cu-N-cyclohexyldiazenium dioxide) component of a wood preservative by a defined soil bacterial community

The wood protection industry has refined their products from chrome-, copper-, and arsenate-based wood preservatives toward solely copper-based preservatives in combination with organic biocides. One of these is Cu-HDO, containing the chelation product of copper and N-cyclohexyldiazenium dioxide (HDO). In this study, the fate of isotope-labeled ((13)C) and nonlabeled ((12)C) Cu-HDO incorporated in wood sawdust mixed with soil was investigated. HDO concentration was monitored by high-pressure liquid chromatography. The total carbon and the δ(13)C content of respired CO(2), as well as of the soil-wood-sawdust mixture, were determined with an elemental analyzer-isotopic ratio mass spectrometer. The concentration of HDO decreased significantly after 105 days of incubation, and after 24 days the (13)CO(2) concentration respired from soil increased steadily to a maximum after 64 days of incubation. Phospholipid fatty acid-stable isotope probing (PFA-SIP) analysis revealed that the dominant PFAs C(19:0)d8,9, C(18:0), C(18:1)ω7, C(18:2)ω6,9, C(17:1)d7,8, C(16:0), and C(16:1)ω7 were highly enriched in their δ(13)C content. Moreover, RNA-SIP identified members of the phylum Acidobacteria and the genera Phenylobacterium and Comamonas that were assimilating carbon from HDO exclusively. Cu-HDO as part of a wood preservative effectively decreased fungal wood decay and overall microbial respiration from soil. In turn, a defined bacterial community was stimulated that was able to metabolize HDO completely.     

东祁连山退化高寒草地土壤细菌群落与土壤环境因子间的相互关系

为探究东祁连山不同退化高寒草地细菌群落分布特征与土壤环境因子间的相互关系,采用高通量测序技术对轻度、中度和重度退化草地的土壤细菌群落结构变化及其多样性进行分析,运用CANOCO 4.5软件对土壤细菌群落与土壤环境因子间关系进行冗余分析(RDA).结果表明:不同退化高寒草地土壤理化性质间均差异显著,高通量测序共得到257125条有效序列,180826条优质序列,4790个OTUs.细菌群落Chao1指数依次为轻度>中度>重度;Shannon指数依次为轻度>重度>中度.系统发育分析表明,各样地土壤细菌类群分属于33个门,其中放线菌门、变形菌门和厚壁菌门是3种不同退化草地土壤中的优势类群.对不同退化草地土壤细菌各门所占比例分析发现,放线菌门、酸杆菌门和变形菌门随着退化程度加剧先减少后增加,厚壁菌门反之.RDA分析结果显示,细菌优势类群与蔗糖酶、纤维素酶和磷酸酶呈极显著相关,与pH、电导率、速效氮、速效钾呈显著相关.说明东祁连山不同退化高寒草地土壤细菌群落间差异明显,土壤环境因子是影响土壤细菌群落分布的重要因素.     

Biodegradation potential of oily sludge by pure and mixed bacterial cultures

The biodegradation capacity of aliphatic and aromatic hydrocarbons of petrochemical oily sludge in liquid medium by a bacterial consortium and five pure bacterial cultures was analyzed. Three bacteria isolated from petrochemical oily sludge, identified as Stenotrophomonas acidaminiphila, Bacillus megaterium and Bacillus cibi, and two bacteria isolated from a soil contaminated by petrochemical waste, identified as Pseudomonas aeruginosa and Bacillus cereus demonstrated efficiency in oily sludge degradation when cultivated during 40 days. The bacterial consortium demonstrated an excellent oily sludge degradation capacity, reducing 90.7% of the aliphatic fraction and 51.8% of the aromatic fraction, as well as biosurfactant production capacity, achieving 39.4% reduction of surface tension of the culture medium and an emulsifying activity of 55.1%. The results indicated that the bacterial consortium has potential to be applied in bioremediation of petrochemical oily sludge contaminated environments, favoring the reduction of environmental passives and increasing industrial productivity.     

Biodegradation of bilge waste from Patagonia with an indigenous microbial community

Oily residues that are generated in normal ship operation are considered hazardous wastes. A biodegradation assay with autochthonous microbiota of Bilge Waste Oily Phase (BWOP) was performed in a bioreactor under controlled conditions. Petroleum, diesel oil, and PAH degraders were isolated from bilge wastes. These bacteria belong to the genus Pseudomonas and are closely related to Pseudomonas stutzeri as shown by 16S rDNA phylogenetic analysis. The indigenous microbial community of the bilge waste was capable of biodegrading the BWOP (1% v/v) with biodegradation efficiencies of 70% for hexane extractable material (HEM), 68% for total hydrocarbons (TH) and 90% for total aromatics hydrocarbons (TA) in 14 days. Solid phase microextraction (SPME) was successfully applied to evaluate hydrocarbon evaporation in a control experiment and demonstrated a mass balance closure of 88%. The SPME and biodegradation results give useful information to improve and scale up the process for BWOP treatment.