Co-metabolic and biochar-promoted biodegradation of mixed PAHs by highly efficient microbial consortium QY1

Polycyclic aromatic hydrocarbons (PAHs), typical representatives of the persistent organic pollutants (POPs), have become ubiquitous in the environment. In this study, a novel microbial consortium QY1 that performed outstanding PAHs-degrading capacity has been enriched. The degradation characteristics of single and mixed PAHs treated with QY1 were studied, and the effect of biochar on biodegradation of mixed PAHs and the potential of biochar in PAHs-heavy metal combined pollution bioremediation were also investigated. Results showed that, in single substrate system, QY1 degraded 94.5% of 500 mg/L phenanthrene (PHE) and 17.8% of 10 mg/L pyrene (PYR) after 7 days, while in PHE-PYR mixture system, the biodegradation efficiencies of PHE (500 mg/L) and PYR (10 mg/L) reached 94.0% and 96.2%, respectively, since PHE served as co-metabolic substrate to have significantly improved PYR biodegradation. Notably, with the cooperation of biochar, the biodegradations of PHE and PYR were greatly accelerated. Further, biochar could reduce the adverse impact of heavy metals (Cd²⁺, Cu²⁺, Cr₂O₇^2?) on PYR biodegradation remarkably. The sequencing analysis revealed that Methylobacterium, Burkholderia and Stenotrophomonas were the dominant genera of QY1 in almost all treatments, indicating that these genera might play key roles in PAHs biodegradation. Overall, this study provided new insights into the efficient bioremediation of PAHs-contaminated site.     

嗜盐菌群对菲的降解及萘双加氧酶基因的表达规律

从胜利油田富集了一个能够降解多环芳烃的嗜盐菌群,通过克隆文库技术解析了菌群萘双加氧酶(ndo)基因的多样性.结果表明,该嗜盐菌群有6种ndo基因型,其中3种主要基因型(占总克隆子92.7%)与经典nah-like基因的相似度为89%.采用real-time RT-PCR等技术分析了不同盐度下菲降解过程中ndo基因的表达量、降解速率、生长曲线以及菲的生物可利用度,探索了菌群对菲的降解及ndo基因的表达规律.结果表明,当盐度由10%升高到20%,菌群完全降解100mg/L菲的时间从6d延长到9d,菌群生长的迟滞期由1d延长为3d.溶解菲的浓度在菌群生长过程中呈增加的趋势.Ndo基因的表达量在降解过程中呈先降低后随溶解菲浓度升高而升高的趋势,表明高浓度菲在初始阶段可能抑制ndo基因的表达.单因素方差分析(n=3, P>0.05)表明溶解菲浓度在10%和20%盐度下没有显著差异,表明一定范围的盐度不会影响菌群降解过程中菲的溶解度.菌群ndo基因的相对表达量在10%盐度下较在20%盐度下高,说明盐度对嗜盐菌群功能基因的表达具有抑制作用.     

高效木薯渣分解复合菌群RXS的构建及其发酵特性研究

从富含腐烂纤维质的环境中取样,通过以木薯渣及滤纸为碳源的蛋白胨纤维素培养基不断地富集培养,构建了一组高效稳定的纤维质分解复合菌群.考察了该复合菌群对不同纤维质底物的分解性能及其在木薯渣水解过程中主要参数的变化.研究发现该复合菌群对滤纸、脱脂棉、微晶纤维素、麦秸秆和木薯渣等原料均能够进行有效的降解.在该复合菌群应用于木薯渣的水解过程中,监测发现纤维素酶、半纤维素酶、果胶酶等关键酶的酶活力分别在第2～3 d达到最大值34.4、90.5和15.8U;经过10 d的发酵后,木薯渣中的纤维素、半纤维素及木质素分别降解了79.8%、85.9%和19.4%,且木薯渣的失重高达61.5%;此外,代谢产物主要是乙酸、丁酸、己酸和甘油;而溶解性COD、总糖和总挥发酸的变化表明第2 d时木薯渣的水解率最高.上述结果表明,该复合菌群能够有效地水解木薯燃料酒精生产过程中的废弃物木薯渣,并有望用于木薯渣高效沼气发酵的前处理中.     

Screening and degrading characteristics and community structure of a high molecular weight polycyclic aromatic hydrocarbon-degrading bacterial consortium from contaminated soil

Inoculation with effcient microbes had been proved to be the most important way for the bioremediation of polluted environments. For the treatment of abandoned site of Beijing Coking Chemical Plant contaminated with high level of high-molecular-weight polycyclic aromatic hydrocarbons(HMW-PAHs) ,a bacterial consortium capable of degrading HMW-PAHs,designated 1-18-1,was enriched and screened from HMW-PAHs contaminated soil.Its degrading ability was analyzed by high performance liquid chromatography(HPLC) ,and the community structure was investigated by construction and analyses of the 16S rRNA gene clone libraries(A,B and F) at different transfers.The results indicated that 1-18-1 was able to utilize pyrene,fluoranthene and benzoapyrene as sole carbon and energy source for growth.The degradation rate of pyrene and fluoranthene reached 82.8%and 96.2%after incubation for 8 days at 30°C,respectively;while the degradation rate of benzoapyrene was only 65.1%after incubation for 28 days at 30°C. Totally,108,100 and 100 valid clones were randomly selected and sequenced from the libraries A,B,and F.Phylogenetic analyses showed that all the clones could be divided into 5 groups,Bacteroidetes,α-Proteobacteria,Actinobacteria,β-Proteobacteria andγ-Proteobacteria.Sequence similarity analyses showed total 39 operational taxonomic units(OTUs) in the libraries.The predominant bacterial groups wereα-Proteobacteria(19 OTUs,48.7%) ,γ-Proteobacteria(9 OTUs,23.1%) andβ-Proteobacteria(8 OTUs,20.5%) . During the transfer process,the proportions ofα-Proteobacteria andβ-Proteobacteria increased greatly(from 47%to 93%) ,while γ-Proteobacteria decreased from 32%(library A) to 6%(library F) ;and Bacteroidetes group disappeared in libraries B and F.     

Phosphate removal using aerobic bacterial consortium and pure cultures isolated from activated sludge

In the present study, an attempt is made to evaluate the kinetics of biological phosphate removal using a bacterial consortium of activated sludge, as well as screening for dominant polyphosphates accumulating bacteria. The results showed an efficient phosphate uptake (P < 0.001) of the consortium, with rates related to the initial concentration of both phosphate and carbon sources. Short chain volatile fatty acids presented the suitable substrates for enhanced biological phosphorus removal, of which maximum yield reached 99.23% and 78.51% in basal salt medium supplemented with 0.5% of sodium acetate and lactate respectively. Fifteen phosphate-accumulating bacteria were isolated from the activated sludge and only four isolates were selected and characterized as Pseudomonas aeruginosa AS1, Moraxella lacunata AS2, Acinetobater junii AS3 and Alcaligenes denitrificans AS4. The highest efficiency of phosphate uptake using pure culture was achieved with Ac. junii AS3 (83.36) followed by P. aeruginosa AS1 (81.78%), Al. denitrificans AS4 (76.72%), and M. lacunata AS2 with 50.6%.     

Isolation and characterization of gasoline-degrading bacteria from gas station leaking-contaminated soils

The effects of culture conditions in vitro and biosurfactant detection were studied on bacterial strains capable of degrading gasoline from contaminated soils near gas station. The main results were summarized as follows. Three bacteria （strains Q10, Q14 and Q18） that were considered as efficiently degrading strains were isolated and identified as Pseudomonas sp., Flavobaeterium sp. and Rhodococcus sp., respectively. The optimal growth conditions of three bacteria including pH, temperature and the concentration of gasoline were similar. The reduction in surface tension was observed with all the three bacteria, indicating the production of biosurfactant compounds. The value of surface tension reduced by the three strains Q10, Q14 and Q18 was 32.6 mN.m, 12.4 mN. m and 21.9 mN.m, respectively. Strain Q10 could be considered as a potential biosurfactant producer. Gasoline, diesel oil, benzene, toluene, ethylbenzene and xylene （BTEX） could easily be degraded by the three isolates. The consortium was more effective than the individual cultures in degrading added gasoline, diesel oil, and BTEX. These results indicate that these strains have great potential for in situ remediation of soils contaminated by gas station leaking.     

Enrichment of thermophilic and mesophilic microbial consortia for efficient degradation of corn stalk

Six different environmental samples were applied to enrich microbial consortia for efficient degradation of corn stalk,under the thermophilic and mesophilic conditions.The consortium obtained from anaerobic digested sludge under thermophilic condition(TC-Y)had the highest lignocellulose-degrading activity.The CO_2yield was 246.73 m L/g VS in23 days,meanwhile,the maximum CO_2production rate was 15.48 mL/(CO_2·d),which was28.75%and 52.27%higher than that under mesophilic condition,respectively.The peak value of cellulase activity reached 0.105 U/mL,which was at least 34.61%higher than the other groups.In addition,49.5%of corn stalk was degraded in 20 days,moreover,the degradation ratio of cellulose,hemicellulose and lignin can reach 52.76%,62.45%and42.23%,respectively.Microbial consortium structure analysis indicated that the TC-Y contained the phylum of Gemmatimonadetes,Acidobacteria,Chloroflexi,Planctomycetes,Firmicutes,and Proteobacteria.Furthermore,the Pseudoxanthomonas belonging to GammaProteobacteria might be the key bacterial group for the lignocellulose degradation.These results indicated the capability of degrading un-pretreated corn stalk and the potential for further investigation and application of TC-Y.     

Effcient degradation of lube oil by a mixed bacterial consortium

A laboratory study was performed to assess the biodegradation of lube oil in bio-reactor with 304# stainless steel as a biofilm carrier. Among 164 oil degrading bacterial cultures isolated from oil contaminated soil samples, Commaonas acidovorans Px1, Bacillus sp. Px2, Pseudomonas sp. Px3 were selected to prepare a mixed consortium for the study based on the e ciency of lube oil utilization. The percentage of oil degraded by the mixed bacterial consortium decreased slightly from 99% to 97.2% as the concentration of lube oil was increased from 2000 to 10,000 mg/L. The degradation of TDOC (total dissolved organic carbon) showed a similar tendency compared with lube oil removal, which indicated that the intermediates in degradation process hardly accumulated. Selected mixed bacterial consortium showed their edge compared to activated sludge. Scanning electron microscopy (SEM) photos showed that biofilms on stainless steel were robust and with a dimensional framework constructed by EPS (extracellular polymeric substances), which could promote the biodegradation of hydrocarbons. The increase of biofilm followed first-order kinetics with rate of 0.216 g glucose/(cm2 day) in logarithm phase. With analysis of Fourier transform infrared spectroscopy (FT-IR) and gas chromatography-mass spectrometry (GC-MS) combined with removal of lube oil and TDOC, mixed bacterial consortium could degrade benzene and its derivatives, aromatic ring organic matters with a percentage over 97%.     

两种N源对石油烃降解菌群降解柴油能力的影响

分别以酵母膏作为有机N源,(NH4)2SO4作为无机N源,在含N量相同(70 mmol/L)的条件下,按接种量2%,柴油1%,25℃,150 r/min摇床恒温培养,对海洋石油烃降解菌群进行了为期30 d 的液体摇瓶降解实验,通过GC-MS测定和分析,对柴油中正烷烃的降解情况进行了对比研究.结果表明,对柴油含量为1%的培养液,应用这两种不同N源,降解菌群对正烷烃的最终降解效果相同,均能完全降解,但有机N源只需3 d 时间就能完全降解正烷烃,比无机N源的降解时间短10倍多.因此,酵母膏对柴油的微生物降解具有极大的促进作用.     

Differential fate of metabolism of a disperse dye by microorganisms Galactomyces geotrichum and Brevibacillus laterosporus and their consortium GG-BL

The present work aims to evaluate Brown 3 REL degrading potential of developed microbial consortium GG-BL using two microbial cultures, Galactomyces geotrichum MTCC 1360 (GG) and Brevibacillus laterosporus MTCC 2298 (BL). Microbial consortium GG-BL showed 100% decolorization of a dye Brown 3 REL, while individually G. geotrichum MTCC 1360 and B. laterosporus MTCC 2298 showed 26% and 86% decolorization under aerobic condition (shaking) respectively. Measurements of biochemical oxygen demand (BOD) (76%) and chemical oxygen demand (COD) (68%) were done after decolorization by consortium GG-BL. No induction in activities of oxidoreductive enzymes found in G. geotrichum while B. laterosporus showed induction of veratryl alcohol oxidase, Nicotineamide adenine dinucleotide-dichlorophenol indophenol (NADH-DCIP) reductase and riboflavin reductase indicating their role in dye metabolism. Consortium GG-BL showed induction in the activities of laccase, veratryl alcohol oxidase, tyrosinase, NADH-DCIP reductase and riboflavin reductase. Two different sets of induced enzymes from G. geotrichum and B. laterosporus work together in consortium GG-BL resulting in faster degradation of dye. The degradation of Brown 3 REL was analyzed using high performance thin layer chromatography (HPTLC), high performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FT-IR) and gas chromatography mass spectroscopy (GC-MS). Phytotoxicity study revealed that metabolites formed after degradation was significantly less toxic in nature.