Biodegradation of high molecular weight PAHs using isolated yeast mixtures: application of meta-genomic methods for community structure analyses

Bioaugmentation for the removal of polyaromatic hydrocarbons (PAHs) from wastewater using bacteria and yeasts is considered environment-friendly and a cost-effective technique. The effectiveness of this biodegradation system depends on the stability of inoculated microorganisms and the availability of nutrients. This study is aimed to investigate the removal of high molecular weight (HMW)-PAHs from biologically treated produced water using different biological systems. Three systems, inoculated with activated sludge (AS), the mixture of five yeast strains (MY), and the mixture of AS and the five yeast strains (SY), respectively, were constructed, and their performance for the removal of HMW-PAHs was compared over 10 weeks. The effluent of the biologically treated produced water from an oilfield was used as the influent after chrysene and benzo(a)pyrene were spiked as HMW-PAHs. Polymerase chain reaction-based denaturing gradient gel electrophoresis (PCR-DGGE) and fluorescent in situ hybridization (FISH) techniques were used to examine the changes in the structures and abundances of the bacterial and yeast communities in these three systems. Only SY and MY systems were capable to remove chrysene (90.7 % and 98.5 %, respectively) and benzo(a)pyrene (80.7 % and 95.2 %, respectively). PCR-DGGE analysis confirmed that all of the five yeast strains inoculated remained in the SY and MY systems, while FISH results showed that the relative abundance of yeast in the SY and MY systems (10.6 % to 21.9 %, respectively) were significantly higher than AS system (2.3 % to 7.8 %, respectively). The relative abundances of the catechol 2,3-dioxygenase (C23O) indicated that the copy number ratios of benzene ring cleavage gene C23O in the yeast amended systems were much higher than that in the AS system. In this study, all of the three systems were effective in removing the low molecular weight (LMW)-PAHs, while HMW-PAHs including chrysene and benzo(a)pyrene were efficiently removed by MY and SY systems, not by AS system. The high HMW-PAHs removal in the MY and SY bioaugmentation systems possibly attributed to the inoculation of the mixed yeast culture. By combining the PCR-DGGE results with the FISH analyses, it was found that yeast probably consisting mainly of the five inoculated strains inhabited in the two bioaugmentation systems as a dominant population. The relatively higher performance of the SY system might be attributed to the suspended growth type which permitted a more efficient contact between microbial cells and contaminants. The bioaugmentation systems (SY and MY) were successfully established by inoculating with five nonindigenous yeast strains and demonstrated high performance in removal of HMW-PAHs.     

Harvesting of Chlorella sp. using hollow fiber ultrafiltration

Introduction

The suitability of the application of ultrafiltration (UF) to harvest Chlorella sp. from the culture medium was examined. We investigated the effects of two improved UF system, forward air?Cwater flushing and backwash with permeate, on the concentration process.

Materials and methods

Backwash with permeate was selected as an optimization of the improved UF system, which was more effective for permeate flux recovery. Moreover, the hollow fiber UF system by adding periodical backwash with permeate was examined for Chlorella sp. harvesting.

Results and discussion

It was found that Chlorella sp. could be concentrated with high recovery in a lab-scale experiment. An overall algal biomass recovery of above 90% was achieved when the volume concentration factor was 10. For an original biomass of 1.3?±?0.05?g/L, 1?min backwash followed by 20?min forward concentrating was more effective, which resulted in a recovery of 94% and a high average flux of 30.3?L/m²/h. In addition, the algal recovery was highly correlated to the volume concentration factor and the initial biomass. A high concentration factor or a high initial biomass resulted in a low biomass recovery.     

纤维素降解细菌筛选及降解特性分析

基于获得高效纤维素降解细菌的目的,通过LB培养基的培养以及刚果红培养基的筛选,从牛粪堆肥中筛选获得2株高效纤维素降解细菌。经鉴定,分别为枯草芽胞杆菌(Bacillus subtilis)和地衣芽胞杆菌(Bacillus licheniformis)。所筛选得到的菌种具有很高的滤纸降解能力,可在6d内使滤纸剧烈崩溃,振摇成均匀糊状;其中,地衣芽胞杆菌的羧甲基纤维素钠酶活峰值在发酵第4天达到峰值(237U/g)。     

垃圾渗滤液发酵产氢和产甲烷特性研究

以实际垃圾渗滤液作为厌氧发酵基质,研究了初始pH为7.0、中温(37℃)条件下的发酵产氢、产甲烷特性。结果表明,利用垃圾渗滤液作为基质发酵产氢或甲烷时,氢气的最大累积产量为24.33mL(以每克COD计,下同),甲烷的最大累积产量为91.59mL,产氢发酵在初期存在明显的迟滞期,但是产甲烷发酵不存在明显迟滞期;产氢发酵的液相末端产物中含有大量的挥发性有机酸和乙醇,乙醇、乙酸、丁酸质量浓度分别为487.23、1 175.21、1 225.78mg/L,相比产氢发酵,产甲烷发酵的液相末端产物中乙醇、乙酸、丁酸质量浓度均较低,分别为256.38、106.73、107.42mg/L;产甲烷发酵的最终pH是6.32,接近中性,而产氢发酵的最终pH为4.21,呈明显酸性;产甲烷发酵对COD的去除率(41.78%)高于产氢发酵对COD的去除率(32.14%),可能是产氢发酵液相末端产物中的乙酸能被产甲烷菌利用,而被进一步降解。     

改性多壁碳纳米管对水中Cd~（2＋）的去除

为了增加多壁碳纳米管（multiwall carbon nanotubers,MWNTs）对水中Cd2＋的吸附量,使用混酸对多壁碳纳米管进行氧化处理,采用红外光谱进行结果表征,并探讨了吸附时间、pH值和MWNTs的使用量、Cd2＋的浓度及干扰离子对镉离子吸附的影响。结果表明,吸附时间为1.5 h、pH为5.3、吸附效果最佳,随MWNTs量的增加Cd2＋去除量增加,共存的阳离子会降低对Cd2＋的吸附效果,对Cd2＋的吸附符合Longmuir吸附定律。研究同时表明,pH小于2时Cd2＋能容易从碳纳米管上解吸。初步探讨了Cd2＋吸附机制。     

芘高效降解菌ZQ_5的培养条件及无机元素对其降解效率的影响

以从我国最大的石油污水灌区之一——沈抚灌区污染土壤分离到的以芘为惟一碳源、能源生长的高效降解菌株ZQ5为实验材料,通过对菌株ZQ5培养条件的优化,以及采用摇瓶振荡培养方法测定菌株ZQ5对不同浓度芘的降解率,表明：菌株ZQ5在30℃振荡培养16 d后对150 mg/L芘的降解率为90.31%。通过模拟稻田施用N、P和K肥等的土壤环境,探索了无机营养元素对降解菌ZQ5降解能力的影响,发现土壤中混合加入N、P和K无机营养元素的降解率能达到82%以上,比单加某种营养元素对降解菌ZQ5的降解效果好。本研究结果可以指导稻田PAHs的原位生物修复。     

Second-order chlorine decay and trihalomethanes formation in a pilot-scale water distribution systems

It is well known that model-building of chlorine decay in real water distribution systems is difficult because chlorine decay is influenced by many factors (e.g., bulk water demand, pipe-wall demand, piping material, flow velocity, and residence time). In this paper, experiments were run to investigate the kinetic model of chlorine decay and the formation model of trihalomethanes (THMs) in pilot-scale water distribution systems. Experimental results show that the rate constants of chlorine decay, including wall decay and bulk decay, increasing with temperature. Moreover, the kinetic model of chlorine decay and the formation model of THMs describe experiment data of pilot-scale water distribution systems. The effect of different piping material on chlorine decay and THMs formation were also investigated. The rate constants of chlorine decay are ranked in order: stainless steel pipe, ductile iron pipe, and last, polyethelene pipe because wall decay is the largest in stainless steel pipe than that in other piping material. Correspondingly, the rate of THMs formation follows the order of stainless steel pipe, ductile iron pipe, and last, polyethelene pipe because of less chlorine in bulk water reacting with the trihalomethane formation potential (THMFP).     

阴极液及底物浓度对AFB-MFC同步废水处理及产电性能影响

为了提高厌氧流化床微生物燃料电池（AFB-MFC）的性能,并为双室MFC寻找价廉、易得、无污染的阴极液,在曝气量16～24 L/h、温度（35±2）℃、回流量10.2 L/h、阴极底边距阴极室内底部17.3 cm、外电阻250 Ω、水力停留时间（HRT）14.0～14.9 h以及进水pH 7.81～8.37下,研究了阴极液及底物浓度对系统产电及废水处理性能的影响。结果表明,使用缓冲溶液、阳极室出水和自来水作阴极液时,自来水的产电性能最佳,阴极液种类不影响系统有机基质的去除。以自来水为阴极液时,阴极液pH及电导率随运行时间增加而增加,COD去除率为80.11%～89.29%,输出电压及功率密度开始随运行时间增加而增加,之后稳定在440～452 mV和48.40～51.08 mW/m²之间。增加底物浓度对COD去除率影响不大,而输出电压及功率密度随底物浓度增加而下降;底物COD浓度由3 307.09 mg/L增至9 520 mg/L时,COD去除率在85.77%～94.44%之间,输出电压及功率密度则分别由449 mV和50.40 mW/m²下降至406 mV和41.21 mW/m²。自来水作阴极液可避免二次污染及阴极液对阳极室微生物的影响,并得到高的产电能力。     

生物通风技术修复柴油污染土壤的土柱模拟实验

生物通风技术是将土壤气相抽提和生物降解结合起来的原位强迫氧化降解方法,对于修复因地下储油罐泄漏引起的土壤污染具有广阔的应用前景。通过室内土柱模拟柴油泄漏污染土壤,分析了不同历时残余总石油烃(total pe-troleum hydrocarbon,TPH)的平衡分布规律以及土壤中不同深度柴油量、总柴油量的变化。结果表明:(1)各柱残余TPH剖面分布差异的原因受土柱的初始装填情况的影响较大;(2)残余TPH平衡分布曲线呈双峰型的土柱,柴油的去除主要以挥发作用及生物降解作用为主;(3)挥发作用主要是由通风孔隙体积数及土壤含水率来影响的;重力作用则主要是由初始油浓度、土壤含水率、C∶N∶P影响的;除通风方式外,其余4个因素都对生物降解作用有影响;(4)初始油浓度较大,土壤含水率较小的柱8和柱11,生物降解作用最明显,柴油去除效果最好。该成果可为生物通风过程的强化提供理论依据。