Impact of Bioaugmentation with a Consortium of Bacteria on the Remediation of Wastewater-Containing Hydrocarbons (5 pp)

Goals, Scope and Background It has been observed that hydrocarbon treated wastewaters still contain high COD and a number of intermediates. This suggests that the required catabolic gene pool for further degradation might be absent in the system or, that its titer value is not significant enough. By providing the desired catabolic potential, the overall efficiency of the treatment system can be improved. This study aims to demonstrate this concept by bioaugmentation of a lab-scale reactor treating refinery wastewater with a consortium having the capacity to complement the alkB genotype to the available microbial population. Methods Two reactors were set up using activated biomass collected from a refinery treatment plant and operated at a continuous mode for a period of 8 weeks. The feed to both reactors was kept constant. Crude oil was spiked regularly. One reactor was bioaugmented with a consortium previously described for crude oil spill remediation. The efficiency of the bioaugmented reactor was demonstrated by reduced COD. The changes in the microbial population over a period of time were analyzed by RAPD. Catabolic activity of the biomass in both reactors was monitored by PCR. The presence of the catabolic loci was confirmed by Southern Hybridization. Results and Discussion 52.2% removal of COD was observed in the bioaugmented reactor while only 15.1% reduction of COD was observed in the reactor without bioaugmentation. The change in microbial population can be seen from the 4th week, which also corresponds to improved catabolic activity. The presence of the bedA locus was seen in all samples, which indicates the presence of aromatic degraders, but the appearance of the alkB locus, from the 6th week onwards, which was observed only in the samples from the bioaugmented reactor. The results suggest that the gene pool of the bioaugmented reactor has catabolic loci that can degrade accumulated intermediates, thus improving the efficiency of the system. Conclusions In this study, improvement of efficiency of bioremediation was demonstrated by addition of catabolic loci that are responsible for degradation. Bioaugmentation was carried out in biomass that was collected from an ETP (effluent treatment plant) treating hydrocarbon containing wastewater to study the strategies for improvement of the treatment system. Biostimulation, only marginally improved the efficiency, when compared to bioaugmentation. The improved efficiency was demonstrated by COD removal. The presence of the alkB locus suggests the importance of a catabolic gene pool that acts on accumulated intermediates. It is well documented that straight chain aliphatics and intermediates of aromatic compounds after ring cleavage, accumulate in refinery wastewater systems, thereby hindering further degradation of the wastewater. Supplementation of a catabolic gene pool that treats the lower pathway compounds and alkanes will improve the overall efficiency. In this study, results suggest that the alkB locus can also be used to monitor the degradative mode of the activated biomass. Recommendations and Perspective . Pollution from petroleum and petroleum products around the globe are known to have grave consequences on the environment. Bioremediation, using activated sludge, is one option for the treatment of such wastes. Effluent treatment plants are usually unable to completely degrade the wastewater being treated in the biological unit (the aerator chambers). The efficiency of degradation can be improved by biostimulation and bioaugmentation. This study demonstrates the improved efficiency of a treatment system for wastewater containing hydrocarbons by bioaugmentation of a consortium that supports degradation. Further experiments on a pilot scale are recommended to assess the use of bioaugmentation on a large scale. The use of molecular tools, like DNA probes for alkB, to monitor the system also needs to be explored.     

水解酸化-厌氧-好氧法处理NF合成制药废水研究

研究了序批式水解酸化 厌氧 好氧生物处理工艺对NF合成制药废水的处理。由于NF制药废水中含有大量有毒有机化合物 ,在生物处理过程中这些有毒物质会抑制活性污泥的活性 ,因此需经过适当稀释原水以达到处理单元可接受的毒性范围。采用BODTrack快速测定了不同原水稀释条件下活性污泥呼吸曲线第一段斜率的变化 ,结果表明 ,当原水稀释 2 0倍以上后 ,对活性污泥的活性没有明显的抑制。通过批量实验 ,优化了工艺的运行条件 ,并进行了小试的连续运行。采用本工艺可以达到NF制药废水COD的稳定高效去除 ,结果显示 ,COD的去除率可达 76 %。     

Naphthalene metabolism in Nocardia otitidiscaviarum strain TSH1, a moderately thermophilic microorganism

The thermophilic bacterium Nocardia otitidiscaviarum strain TSH1, originally isolated in our laboratory from a petroindustrial wastewater contaminated soil in Iran, grows at 50 degrees C on a broad range of hydrocarbons. Transformation of naphthalene by strain TSH1 which is able to use this two ring-polycyclic aromatic hydrocarbon (PAH) as a sole source of carbon and energy was investigated. The metabolic pathway was elucidated by identifying metabolites, biotransformation studies and monitoring enzyme activities in cell-free extracts. The identification of metabolites suggests that strain TSH1 initiates its attack on naphthalene by dioxygenation at its C-1 and C-2 positions to give 1,2-dihydro-1,2-dihydroxynaphthalene. The intermediate 2-hydroxycinnamic acid, characteristic of the meta-cleavage of the resulting diol was identified in the acidic extract. Apart from typical metabolites of naphthalene degradation known from mesophiles, benzoic acid was identified as an intermediate for the naphthalene pathway of this Nocardia strain. Neither phthalic acid nor salicylic acid metabolites were detected in culture extracts. Enzymatic experiments with cell extract showed the catechol 1,2-dioxygenase activity while transformation of phthalic acid and protocatechuic acid was not observed. The results of enzyme activity assays and identification of benzoic acid in culture extract provide strong indications that further degradation goes through benzoate and beta-ketoadipate pathway. Our results indicate that naphthalene degradation by thermophilic N. otitidiscaviarum strain TSH1 differs from the known pathways found for the thermophilic Bacillus thermoleovorans Hamburg 2 and mesophilic bacteria.     

MFC-MBR耦合系统运行效果

膜生物反应器(MBR)是一种高效的污水处理工艺,而微生物燃料电池(MFC)能有效降解污泥中的胞外生物有机质(EBOM)并回收电能.将MFC与MBR联用,建立了一套能够有效抑制膜污染同时回收电能的新系统——MFC-MBR耦合系统,MBR的剩余污泥经MFC处理后回流.以传统MBR为对照,对耦合系统中污水处理效果、膜污染情况和污泥混合液的性质进行研究.研究表明,耦合系统的污水处理效果没有明显恶化,COD去除率为94％,NH4+-N的去除率为92％.耦合系统能够有效减缓膜污染的发生,清洗周期延长了28％.污泥混合液的MLVSS/MLSS稳定在80％ ～ 88％,系统内几乎没有无机颗粒积累.松散结合态胞外聚合物(LB-EPS)降低了48％,使污泥混合液性质得到改善.较低的污泥比阻(2.69×1012m/kg)和标准化毛细吸水时间(1.67 s·L/g MLSS),证明耦合系统污泥混合液脱水性能提高了.     

Molecular perspectives and recent advances in microbial remediation of persistent organic pollutants

Nutrition and pollution stress stimulate genetic adaptation in microorganisms and assist in evolution of diverse metabolic pathways for their survival on several complex organic compounds. Persistent organic pollutants (POPs) are highly lipophilic in nature and cause adverse effects to the environment and human health by biomagnification through the food chain. Diverse microorganisms, harboring numerous plasmids and catabolic genes, acclimatize to these environmentally unfavorable conditions by gene duplication, mutational drift, hypermutation, and recombination. Genetic aspects of some major POP catabolic genes such as biphenyl dioxygenase (bph), DDT 2,3-dioxygenase, and angular dioxygenase assist in degradation of biphenyl, organochlorine pesticides, and dioxins/furans, respectively. Microbial metagenome constitutes the largest genetic reservoir with miscellaneous enzymatic activities implicated in degradation. To tap the metabolic potential of microorganisms, recent techniques like sequence and function-based screening and substrate-induced gene expression are proficient in tracing out novel catabolic genes from the entire metagenome for utilization in enhanced biodegradation. The major endeavor of today’s scientific world is to characterize the exact genetic mechanisms of microbes for bioremediation of these toxic compounds by excavating into the uncultured plethora. This review entails the effect of POPs on the environment and involvement of microbial catabolic genes for their removal with the advanced techniques of bioremediation.     

活性污泥数学模型中异养菌产率系数的测定

活性污泥数学模型常被用于预测污水处理厂中的生物过程。模型中异养菌产率系数YH对其他组分的测定值有显著的影响。目前．测定YH的主要方法有序批式活性污泥法和序批式呼吸计量法。在序批式呼吸计量法中．起始F／M值的选择十分关键。以醋酸钠为底物．对这两种方法进行了分析比较,序批式呼吸计量法为文章的推荐方法。     

Effect of alkyl hydroxybenzenes on the properties of dioxygenases

The aim of the present work was to investigate the influence of alkylhydroxybenzenes (AHBs) and tyrosol, which belong to cell differentiation factors d(1) group of autoregulators on properties of biodegradation enzymes, catechol 1,2-dioxygenase (Cat 1,2-DO) and methylcatechol 1,2-dioxygenase (MCat 1,2-DO) of Rhodococcus opacus 6a. AHBs were found to have a greater effect on MCat 1,2-DO than on Cat 1,2-DO. It was expressed by more pronounced changes in the activity of MCat 1,2-DO with unsubstituted catechol at different AHB concentrations and by increasing thermostability of MCat 1,2-DO compared to Cat 1,2-DO under the protective action of AHBs. The compound C(7)-AHB shifted the maximum of dioxygenase activities towards higher temperatures and increased their operation optimum. AHBs changed the specificity constant of dioxygenases by decreasing/increasing the K(m)/V(max) value. For example, the increase in the V(max) value of 3,6-dichlorocatechol oxidation by Cat 1,2-DO in the presence of C(7)-AHB was 300-fold higher compared to the same reaction without AHB. The influence of cell differentiation factors on the properties of dimeric enzymes has been shown for the first time. It gives an idea of how the specificity of enzymes can be changed in vivo when strains contact new substrates. The work has shown the possibility of modification of the properties of dimeric enzymes towards the extension of enzyme activity with difficulty converted substrates or in more extreme conditions, which may be important for biotechnological processes.     

Benzoate degradation by Rhodococcus opacus 1CP after dormancy: Characterization of dioxygenases involved in the process

The process of benzoate degradation by strain Rhodococcus opacus 1CP after a five-year dormancy was investigated and its peculiarities were revealed. The strain was shown to be capable of growth on benzoate at a concentration of up to 10 g L^?1. The substrate specificity of benzoate dioxygenase (BDO) during the culture growth on a medium with a low (200–250 mg L^?1) and high (4 g L^?1) concentration of benzoate was assessed. BDO of R. opacus 1CP was shown to be an extremely narrow specificity enzyme. Out of 31 substituted benzoates, only with one, 3-chlorobenzoate, its activity was higher than 9% of that of benzoate. Two dioxygenases, catechol 1,2-dioxygenase (Cat 1,2-DO) and protocatechuate 3,4-dioxygenase (PCA 3,4-DO), were identified in a cell-free extract, purified and characterized. The substrate specificity of Cat 1,2-DO isolated from cells of strain 1CP after the dormancy was found to differ significantly from that of Cat 1,2-DO isolated earlier from cells of this strain grown on benzoate. By its substrate specificity, the described Cat 1,2-DO was close to the Cat 1,2-DO from strain 1CP grown on 4-methylbenzoate. Neither activity nor inhibition by protocatechuate was observed during the reaction of Cat 1,2-DO with catechol, and catechol had no inhibitory effect on the reaction of PCA 3,4-DO with protocatechuate.     

Estimation of biodegradation parameters of phenolic compounds on activated sludge by respirometry

The bio-oxidation of phenol, catechol, resorcinol, m-cresol and 5-methylresorcinol on activated sludge was investigated by oxygen uptake measurements. In addition, the degradation of acetate with the same microbial population was studied. The substrate-dependent oxygen uptake data were analysed on the basis of the Michaelis-Menten kinetics. The extant kinetic parameters, the maximum rates of oxygen consumption and half-saturation constants for the processes with different substrates were determined. The simple respirometric approach also made it possible to determine the short-term oxygen demands of the substrates which formed 23-38% of the theoretical oxygen demand of the studied compounds.     

污泥淤砂分离器再分离底流污泥

研究了污泥淤砂分离器对分离原污泥所得的底流污泥进行再分离时,分离分流污泥的性质。实验结果表明,原污泥MLVSS/MLSS为0.372,1次分离后得到的底流污泥MLVSS/MLSS为0.222,2次分离后得到的底流污泥MLVSS/MLSS为0.126,表明通过底流污泥再分离,能够进一步降低底流污泥中生物有机质的含量,提高底流污泥中无机淤砂的含量。同时,原污泥CST为2.85(s·L)/g SS,1次分离后得到的底流污泥CST为0.98(s·L)/g SS,2次分离后得到的底流污泥CST为0.12(s·L)/g SS,表明底流污泥再分离进一步提高了底流污泥的脱水性能,最终得到的底流污泥脱水性能良好。     

膜-生物反应器处理超市废水中污泥性质与膜污染关系分析

在以处理超市废水实际工程的基础上,通过对污泥性质中的胞外聚合物(EPS)、溶解性微生物产物(SMPs)、溶解性COD(SCOD)和污泥浓度(MLSS)变化的分析,得出EPS积累的原因是多方面的,而EPS的过度积累对沉降性能有一定的恶化作用;此外,EPS与SMPs有很好的相关性.作者采用SPSS软件对污泥性质与膜污染之间...     

Identifying different types of bulking in an activated sludge system through quantitative image analysis

The present study proposes an image analysis methodology for the identification of different types of disturbances in wastewater treatment activated sludge systems. Up to date, most reported image analysis methodologies have been used in activated sludge processes with the aim of filamentous bulking detection, however, other disturbances could be foreseen in wastewater treatment plants. Such disturbances can lead to fluctuations in the biomass contents, affecting the mixed liquor suspended solids (MLSS), and in the sludge settling ability, affecting the sludge volume index (SVI). Therefore, this work focuses on predicting the MLSS and SVI parameters for different types of disturbances affecting an activated sludge system. Four experiments were conducted simulating filamentous bulking, zoogleal or viscous bulking, pinpoint floc formation, and normal operating conditions. Alongside the MLSS and SVI determination, the aggregated and filamentous biomass contents and morphology were studied as well as the biomass Gram and viability status, by means of image analysis.     

污泥淤砂分离器再分离溢流污泥

研究了污泥淤砂分离器对分离原污泥所得的溢流污泥进行再分离时,分离器的分离效果和分离分流污泥的性质。实验结果表明,污泥淤砂分离器对溢流污泥进行再分离时,分离器的除砂效率为34.5%,分离度为1.60,与分离原污泥时相比,分离器的分离性能降低;分离得到的溢流污泥MLVSS/MLSS比值为0.406,仅比进料污泥增加了2.2%,未能进一步提高溢流污泥MLVSS/MLSS比值;分离得到的底流污泥浓度MLSS为16.81 g/L,仅为原污泥浓度的1.4倍,底流污泥SVI、CST分别为45.7 mL/g和1.37 s·L/g SS,与原污泥相比仅分别减小了9.3%和2.7%,溢流污泥再分离,会降低底流污泥的浓缩效果,降低底流污泥沉降性能、脱水性能的提高幅度,不利于底流污泥的处理处置。     

地下水浅埋区某加油站特征污染物空间分布简

加油站渗漏污染地下水已经是一个世界性的问题。由于浅埋区加油站储罐与地下水密切接触,更加剧储罐的腐蚀。为揭示加油站渗漏的典型污染物石油烃（TPH）、苯系物（BTEX）、萘和甲基叔丁基醚（MTBE）在该水文地质条件下的迁移变化,在浅埋区某加油站开展了平、枯、丰水期的地下水监测工作。在水平分布上,TPH、BTEX、萘基本相似,均在加油岛附近形成高浓度区,而MTBE则更易随地下水流动而迁移,呈现出不同的污染晕。在垂直分布上,地下水的水位变动是污染物浓度分布的主要影响因素。     

红斑体虫的污泥减量效果

为了实现红斑顠体虫捕食污泥减量化,对不同条件下红斑顠体虫的污泥减量效果进行实验研究。实验结果表明:红斑顠体虫的污泥减量速率随初始MLSS及温度的增大而增大,初始MLSS越高,污泥减量速率越大。污泥减量速率随红斑顠体虫密度变化率的增大而逐渐增大,当红斑顠体虫的密度增长率出现下降时,污泥减量速率也呈下降趋势。采用间歇曝气(12 h曝气,12 h停曝)方式,红斑顠体虫的污泥减量速率会显著下降。在污泥好氧消化时,红斑顠体虫能捕食污泥中的有机碎片和细菌,达到污泥稳定化的指标要求。     

红斑顠体虫的污泥减量效果

为了实现红斑顠体虫捕食污泥减量化,对不同条件下红斑顠体虫的污泥减量效果进行实验研究。实验结果表明：红斑顠体虫的污泥减量速率随初始MLSS及温度的增大而增大,初始MLSS越高,污泥减量速率越大。污泥减量速率随红斑顠体虫密度变化率的增大而逐渐增大,当红斑顠体虫的密度增长率出现下降时,污泥减量速率也呈下降趋势。采用间歇曝气（12 h曝气,12 h停曝）方式,红斑顠体虫的污泥减量速率会显著下降。在污泥好氧消化时,红斑顠体虫能捕食污泥中的有机碎片和细菌,达到污泥稳定化的指标要求。     

Aerobic degradation of di- and trichlorobenzenes by two bacteria isolated from polluted tropical soils

Two polychlorinated biphenyl (PCBs)-degrading bacteria were isolated by traditional enrichment technique from electrical transformer fluid (Askarel)-contaminated soils in Lagos, Nigeria. They were classified and identified as Enterobacter sp. SA-2 and Pseudomonas sp. SA-6 on the basis of 16S rRNA gene analysis, in addition to standard cultural and biochemical techniques. The strains were able to grow extensively on dichloro- and trichlorobenzenes. Although they failed to grow on tetrachlorobenzenes, monochloro- and dichlorobenzoic acids, they were able to utilize all monochlorobiphenyls, and some dichlorobiphenyls as sole sources of carbon and energy. The effect of incubation with axenic cultures on the degradation of 0.9 mM 1,4-dichlorobenzene, 0.44 mM 1,2,3- and 0.43 mM 1,3,5-trichlorobenzene in mineral salts medium was studied. Approximately, 80-90% of these xenobiotics were degraded in 200 h, concomitant with cell increase of up to three orders of magnitude, while generation times ranged significantly (P<0.05) from 17-32 h. Catechol 1,2-dioxygenase and catechol 2,3-dioxygenase activities were detected in crude cell-free extracts of cultures pre-grown with benzoate, with the latter enzyme exhibiting a slightly higher activity (0.15-0.17 micromolmin(-1) mg of protein(-1)) with catechol, suggesting that the meta-cleavage pathway is the most readily available catabolic route in the SA strains. The wider substrate specificity of these tropical isolates may help in assessing natural detoxification processes and in designing bioremediation and bioaugmentation methods.     

活性污泥法处理高钙废水中污泥特性的变化

通过单级SBR法处理模拟高钙废水,研究了活性污泥法处理高钙废水的过程中钙离子对COD,MLVSS,MLSS,SVI,污泥增长速率,污泥形态结构及生物相的影响,揭示活性污泥法处理高钙废水的过程中污泥量巨大的原因。采用逐步增加钙离子浓度的方法,检测到在污泥培养期([Ca2+]=0 mg/L),COD去除率为98.1%,MLVSS和MLSS稳定在4 900~5 500mg/L,污泥增长速率为67 mg/(L·d),SVI为55~60 mL/g;在驯化处理期([Ca2+]=120~2 400 mg/L),COD去除率降至87.37%,MLVSS降至2 500 mg/L,MLSS增加至19 300 mg/L,污泥增长速率为212.31 mg/(L·d),SVI降至25 mL/g;在冲击期([Ca2+]=4 000 mg/L),COD去除率降至69.23%,MLVSS降至1 600 mg/L,MLSS迅速增加至24 200 mg/L,污泥增长速率为816.67 mg/(L·d),SVI降至14 mL/g。经显微镜观察发现,污泥絮体由松散变得密实,生物相由钟虫等指示性微生物变为不适应环境的胞囊结构。结果表明,随Ca2+浓度的增加,COD去除率下降,MLSS迅速增加,MLVSS和SVI急剧缩小,说明活性污泥中的活性微生物逐渐减少,而无机物组分逐渐增多;钙离子的加入促使系统碳酸平衡向右移动,使离子状态的钙大部分转化为难降解的碳酸盐,并附着于污泥絮体上,污泥绒粒被压缩,使污泥颗粒密实度及MLSS迅速增加,导致污泥排放量巨大。     

生物法降解养殖场臭气中H_2S的反应器启动

畜禽养殖场臭气成分复杂,完全去除较为困难。生物法是目前应用较广泛的脱臭方法,其中能否将生物膜附着在填料上是影响生物法去除恶臭气体效率的重要因素。本实验采用定时定量投加Na2S的方式驯化活性污泥,并选用MLSS浓度和SO42-浓度增量变化2个指标作为污泥驯化成熟的指标,比传统的以MLSS作为污泥驯化成熟的指标更准确。采用循环污泥的挂膜方式,运行2 d后,通入新鲜的空气和H2S气体,2周后反应器启动成功。     

污泥脱水性能的关键影响因素研究

针对目前污泥脱水性能影响因素不明的问题,研究通过对SBR在7种运行工况条件下活性污泥胞外聚合物(extracellular polymeric substances,EPS)的提取和测试,并采用SPSS软件分析,探讨MLVSS/MLSS、胞外蛋白质/胞外多糖、胞外蛋白质含量、胞外多糖含量、EPS总含量对污泥脱水性能的影响。研究结果表明,MLVSS/MLSS、胞外蛋白质/胞外多糖是影响污泥脱水性能的关键因素,并建立了脱水污泥含水率和污泥结合水含量的影响因素模型:脱水污泥含水率=0.229×(MLVSS/MLSS)+0.023×(胞外蛋白质/胞外多糖)+0.539、污泥结合水含量=2.876×(MLVSS/MLSS)+0.335×(胞外蛋白质/胞外多糖)-0.012,通过对7个城市污水厂污泥脱水性能测试比较表明,模型对脱水污泥含水率和污泥结合水含量预测值相对误差分别在±1%和±5%以内,对污泥脱水性能预测效果良好。