Mass transfer and bioremediation of naphthalene particles in a roller bioreactor

Naphthalene particles in a water slurry have been bioremediated in a sealed, roller bioreactor using a pure strain of Pseudomonas putida. High stripping losses of particles due to both splashing and aeration made the use of the traditional CSTR bioreactor unsuitable for bioremediation of naphthalene particles. The overall dissolution mass transfer coefficient of naphthalene particles in the roller bioreactor was low, 0.055 h(-1) at 50 RPM. The dissolution mass transfer rate was the limiting step for bioremediation. Although mass transfer was identified as the rate limiting step, the addition of hydroxypropyl-beta-cyclodextrin (a solubility enhancer) failed to improve naphthalene slurry bioremediation. In order to successfully bioremediate naphthalene particles at concentrations over 300 mg/L, intermittent aeration was applied in the sealed roller bioreactor on a daily basis. By operating in sequential batch mode with intermittent aeration, the roller bioreactor was successfully used to continuously bioremediate naphthalene particles at concentrations up to 1000 mg/L and at rates up to 10 mg/Lh.     

Bench-scale and field-scale evaluation of catechol 2,3-dioxygenase specific primers for monitoring BTX bioremediation

The objective of this work was to test a molecular genetic method for in situ monitoring of aerobic benzene, toluene, and xylene (BTX) biodegrading microorganisms. Catechol 2,3-dioxygenase (C23DO) genes occur in bacteria that biodegrade benzene, toluene, xylenes, phenol, biphenyl, and naphthalene. A competitive quantitative polymerase chain reaction (QC-PCR) technique using a single set of primers specific for an entire subfamily of C23DO genes was recently developed. To determine whether bacteria containing these C23DO genes actually exist in environments contaminated by BTX, aerobic microcosms containing previously uncontaminated soil were amended with different aromatic hydrocarbons and DNA extracts were analyzed by QC-PCR for C23DO genes. Anaerobic microcosms were established to confirm that oxygen was also necessary for the enrichment of C23DO genes. Field testing was done at two sites undergoing monitored natural attenuation. In microcosm experiments naphthalene, m-xylene, and p-xylene strongly enriched for C23DO genes while benzene, toluene, and o-xylene produced only transient, weakly detectable genes. In the field study, C23DO genes were detected in groundwater samples contaminated with either xylenes or naphthalene. The results of this study demonstrated that molecular genetic techniques can provide an accurate and rapid method to detect microorganisms capable of aromatic hydrocarbon biodegradation. Such a technique would be useful for monitoring the effectiveness of aeration technologies and for documenting microbial processes for monitored natural attenuation.     

纺锤芽孢杆菌降解水中萘的特性研究

从处理石油废水的曝气池污泥中筛选、分离到一株能有效降解萘的菌株,经鉴定为纺锤芽孢杆菌(BFN),研究了其对水中萘的降解特性。结果表明:在温度为30℃、自然pH(6.68～6.76)、接种量为0.2%、(NH4)2SO4浓度为0.15g/L的最适降解条件下,BFN对萘(初始浓度为50mg/L)的降解率在96h内达到99.8%;BFN还具有较好的耐盐度,对高浓度的萘也有较好的耐受性。BFN对萘的降解过程符合一级反应动力学。通过检测不同底物水样的吸光度、pH和底物浓度的变化,发现BFN还能降解苯甲酸、水杨酸、邻苯二甲酸、甲苯、苯酚以及1-萘酚。     

A study of external mass transfer effect on biodegradation of phenol using low‐density polyethylene immobilized Bacillus flexus GS1 IIT (BHU) in a packed bed bioreactor

The impact of external mass transport on the biodegradation rate of phenol in a packed bed bioreactor (PBBR) was studied. A potential bacterial species, Bacillus flexus GS1 IIT (BHU), was isolated from the petroleum‐contaminated soil. Low‐density polyethylene (LDPE) immobilized with the B. flexus GS1 IIT (BHU) was used as packing material in the PBBR. The PBBR was operated by varying the inlet feed flow rate from 4 to 10 mL/min, and the corresponding degradation rate coefficients were found to be in the range of 0.119–0.157 L/g h. In addition, the highest removal rate of phenol was obtained to be 1.305 mg/g h at an inlet feed rate of 10 mL/min. The external mass transfer was studied using the model . A new empirical correlation for the biodegradation of phenol in the PBBR was developed after the evaluation at various values of K and n.     

Rhizoremediation of hydrocarbon contaminated soil using Australian native grasses

Rhizoremediation involves the breakdown of contaminants in soil resulting from microbial activity that is enhanced in the plant root zone. The objective of this study was to assess Australian native grasses for their ability to stimulate removal of aliphatic hydrocarbons from a mine site soil. Time-course pot experiments were conducted in a greenhouse with three grass species (Cymbopogon ambiguus, Brachiaria decumbens, and Microlaena stipoides) in a mine site soil experimentally contaminated with a 60:40 diesel:oil mixture at 1% (w/w) concentration. Plants were cultivated for 100 days with periodic evaluation of changes in soil total petroleum hydrocarbon (TPH) concentration, soil lipase activity, and abundance of hydrocarbon-degrading microorganisms. Results were compared to unplanted control treatments. Significantly lower endpoint TPH concentrations were recorded in planted soil compared to unplanted soil (p = 0.01). Final TPH concentrations and rates of TPH removal varied between grass species, with total TPH removal of between 50% and 88% achieved in planted treatments. The presence of grasses significantly increased the abundance of hydrocarbon-degrading microorganisms and soil lipase activity relative to unplanted soil (p < 0.05). Residual TPH concentration was found to be closely (negatively) correlated with abundance of hydrocarbon-degrading microorganisms and to a lesser extent with soil lipase activity. Australian native grass species were identified that effectively enhance the remediation of diesel/oil contaminated soil, without any requirement for nutrient supplementation. Results may have extensive application to the nationwide problems associated with hydrocarbon contaminated sites.     

Biodegradation of agricultural herbicides in sequencing batch reactors under aerobic or anaerobic conditions

This study investigated the biodegradability of the herbicides isoproturon and 2,4-dichlorophenoxyacetic acid (2,4-D) in sequencing batch reactors (SBRs). Two laboratory-scale (2L liquid volume) SBRs were employed: one reactor performing under aerobic and the other under anaerobic conditions. The aerobic SBR was operated at an ambient temperature (22+/-2 degrees C), while the anaerobic SBR was run in the lower mesophilic range (30+/-2 degrees C). Each bioreactor was seeded with a 3:1 mixture (by weight) of fresh sludge and biomass that had been previously exposed to both herbicides. The effect of herbicide concentration on either treatment process was explored at a hydraulic retention time (HRT) of 48 h, using glucose as a supplemental carbon substrate. Although no isoproturon degradation was observed in either system during the study, complete 2,4-D removal occurred after an acclimation period of approximately 30 d (aerobic SBR) and 70 d (anaerobic SBR). The aerobic reactor achieved complete 2,4-D utilization at feed concentrations up to 500 mg/L. A further increase to 700 mg/L, however, proved to be inhibitory since 2,4-D biodegradation was negligible. On the other hand, the anaerobic SBR was able to degrade 120 mg/L of 2,4-D, which corresponds to 40% of the maximum feed concentration applied. Moreover, glucose was consumed first throughout the experiment in a sequential utilization pattern relating to 2,4-D, with biodegradation of both substrates following closely first-order kinetics.     

Biodegradation of 1,4-dioxane in planted and unplanted soil: effect of bioaugmentation with Amycolata sp. CB1190

1,4-Dioxane is one of the most recalcitrant and toxic contaminants in the subsurface. This study investigated the potential to enhance dioxane biodegradation in both planted and unplanted soil, by adding the dioxane-degrading actinomycete, Amycolata sp. CB1190. Dioxane was not removed within 120 days in sterile controls or in viable microcosms not amended with CB 1190. Poplar root extract (40 mg/L as COD) stimulated dioxane degradation in bioaugmented soil, and 100 mg/L dioxane were removed within 45 days. Other co-substrates that enhanced dioxane degradation by CB1190 include tetrahydrofuran (THF) and 1-butanol, while glucose and soil extract did not affect dioxane degradation. The stimulatory effect of THF was partly due to enhanced enzyme induction, while that of root extract and 1-butanol was attributed to additional growth of CB1190. In another experiment with dioxane added at 10 mg/kg-soil. reactors planted with hybrid poplar trees removed (by evapotranspiration and biodegradation in the root zone) more dioxane within 26 days than unplanted reactors, regardless of whether CB1190 was added. Nevertheless, CB1190 enhanced mineralization of [14C]-dioxane in all experiments. This enhancement was more pronounced in unplanted soil because plant uptake reduced the availability of dioxane for microbial degradation. These results suggest that bioaugmented phytoremediation is an attractive alternative to remove dioxane from shallow contaminated sites.     

Effects of heat-activated persulfate oxidation on soil microorganisms

The effects of heat-activated persulfate on indigenous microorganisms and microcosms augmented with Pseudomonas putida KT2440 were studied in laboratory batch reactors with aquifer material. Microscopic enumeration was used to measure the changes in cell density, and acetate consumption was used to evaluate metabolic activity after exposure to activated persulfate. The cell enumerations showed that persulfate concentrations up to 10 g/L did not affect the indigenous microorganisms but were detrimental to P. putida survival. Acetate consumption was inhibited at the highest persulfate dose (10 g/L). The results emphasize the necessity of using multiple toxicity assays and indigenous cultures in order to realistically assess the potential effects of in situ chemical oxidation on soil microorganisms. A comparison to other studies suggests that the effects of activated persulfate on soil microorganisms are less damaging than those of Fenton's reagent and hydrogen peroxide.     

SPME-GCMS study of the natural attenuation of aviation diesel spilled on the perennial ice cover of Lake Fryxell, Antarctica

In January 2003, a helicopter crashed on the 5 m thick perennial ice cover of Lake Fryxell (McMurdo Dry Valleys, East Antarctica), spilling approximately 730 l of aviation diesel fuel (JP5-AN8 mixture). The molecular composition of the initial fuel was analyzed by solid phase microextraction (SPME) gas chromatography-mass spectrometry (GC-MS), then compared to the composition of the contaminated ice, water, and sediments collected a year after the spill. Evaporation is the major agent of diesel weathering in meltpool waters and in the ice. This process is facilitated by the light non-aqueous phase liquid properties of the aviation diesel and by the net upward movement of the ice as a result of ablation. In contrast, in sediment-bearing ice, biodegradation by both alkane- and aromatic-degraders was the prominent attenuation mechanism. The composition of the diesel contaminant in the ice was also affected by the differential solubility of its constituents, some ice containing water-washed diesel and some ice containing exclusively relatively soluble low molecular weight aromatic hydrocarbons such as alkylbenzene and naphthalene homologues. The extent of evaporation, water washing and biodegradation between sites and at different depths in the ice are evaluated on the basis of molecular ratios and the results of JP5-AN8 diesel evaporation experiment at 4 degrees C. Immediate spread of the aviation diesel was enhanced where the presence of aeolian sediments induced formations of meltpools. However, in absence of melt pools, slow spreading of the diesel is possible through the porous ice and the ice cover aquifer.     

胞外表面活性物质在石油烃生物降解过程中的形成及其作用

采用SBR法处理含葡萄糖和石油烃的人工废水,实验结果表明：在生物处理过程中,微生物首先快速吸附、降解葡萄糖,石油烃类随后也能得到有效降解;在烃类与葡萄糖共存的情况下,微生物在胞外产生更多的蛋白类物质：含石油烃和不合石油烃反应器中污泥胞外蛋白类物质平均含量分别为82．9和58．1mg／gVSS,EPS的乳化指数分别为50％和12．5％,说明胞外的蛋白类物质具有一定的表面活性,可以将烃类在一定程度上乳化,从而有利于微生物对烃类的接触和摄取。     

碳源对好氧颗粒污泥物理性状及除磷性能的影响

采用气提升内循环序批式反应器，分别以葡萄糖、乙酸钠、乙醇为碳源培养好氧颗粒污泥，考察了3种颗粒污泥的物理性状、除磷性能及除磷机理。结果表明：3种颗粒污泥在结构和处理效果上均能长时间保持稳定，其中以乙酸钠和葡萄糖为碳源培养出的颗粒污泥具有较密实的内部结构和较好的沉降性能；在进水COD为600mg／L、TP为15mg／L时，以葡萄糖、乙酸钠、乙醇为碳源培养出的好氧颗粒污泥对TP的去除率分别为82％、88％、52％，其中对TP去除效果较好的两种好氧颗粒污泥（以乙酸钠、葡萄糖为碳源培养的）在反应初期均有较明显的释磷现象发生；以乙酸钠为碳源培养出的颗粒污泥在进水COD浓度为800mg／L时对TP的去除率达到最高（为92％）．进一步加大COD负荷会使好氧颗粒污泥的除磷能力迅速下降。     

阴离子交换膜生物反应器反硝化性能的研究

考察了进水硝酸盐浓度对阴离子交换膜生物反应器反硝化性能的影响。试验结果表明,当进水NO3-为47.01～168.55mg/L时,流动池出水中的NO3-N浓度满足我国生活饮用水水质标准中小于10mg/L的要求(NO3-≤44.29mg/L)。厌氧生物反应器对硝酸盐具有较高的反硝化性能。流动池出水中的Cl-浓度随着进水NO3-浓度的增加而升高,但出水pH值稳定在6.8左右。出水总有机碳浓度与进水的保持一致,表明其未受到"二次污染"和"微生物污染"。     

固定化氨氧化菌短程硝化的启动研究

采用亲水性玻璃态单体,应用辐射技术制备生物相容性高分子共聚物载体,使用固定化细胞增殖技术对氨氧化菌进行固定化,并以流化床为生物反应器,采用SBR运行方式对人工配水进行短程硝化的启动研究.结果表明:当进水氨氮浓度为100、75、50和25mg/L时,对氨氮的的去除率分别为98.6%、99.1%、98.8%和99.8% ,亚硝化率分别为 98.6%、94,5%、95.2%和94.7%:对氨氮的去除速率由开始时的10.6mg/(L·d)提高到25.7mg/(L·d),耗氧速率(OUR)则由0.37mg/(L·d)提高到1.12mg/(L·d).可见,该方法具有启动速度快、亚硝化程度高、容易控制等优点.     

Operational strategies for an activated sludge process in conjunction with ozone oxidation for zero excess sludge production during winter season

A pilot-scale activated sludge system coupled with sludge ozonation process was operated for 112 days of a winter season without excess sludge wasting. The concept of this process is that the excess sludge produced is first disintegrated by ozone oxidation and then recirculated to a bioreactor in order to mineralize the particulate and soluble organic compounds. The basis of operation was to determine either the optimal amount of sludge in kg SS ozonated each day (SO) or the optimal ozonation frequency under the variable influent chemical oxygen demand (COD) loading and temperature conditions, since the ozone supply consumes costly energy. The optimal SO was obtained using the theoretically estimated sludge production rate (SP) and experimentally obtained ozonation frequency (n). While the SP was mainly subject to the COD loadings, sludge concentration was affected by the temperature changes in winter season. The optimal n was observed between 2.5 and 2.7 at around 15 degrees C, but it was doubled at 10 degrees C. Mixed liquor suspended solids (MLSS) concentration was leveled off at around 5000 mg/L in bioreactor at 15 degrees C, but the volatile fraction of MLSS was fixed around 0.7 indicating that there was no significant inorganic accumulation. Suspended solids (SS) and soluble COD in effluents kept always a satisfactory level of 10 and 15 mg/L with sufficient biodegradation. It was recommended to apply a dynamic SO under variable influent COD loadings and temperature conditions to the activated sludge system without excess sludge production for saving energy as well as system stabilization.     

Application of nonionic surfactant-enhanced in situ flushing to a diesel contaminated site

Surfactant-enhanced in situ flushing was performed to remediate soil and groundwater at a diesel contaminated area, which had been used as a military vehicle repair area in Korea for 45 years. A pilot-scale site (4 m x 4 m x 4 m) was selected within the contaminated area for in situ flushing; the selected site was composed of heterogeneous sandy and silt-sandy soils, with an average hydraulic conductivity (K) of 2.0 x 10(-4)cm/s. Two percent of sorbitan monooleate (POE 20) was mixed with uncontaminated groundwater and five pore volumes of solution (three pore volumes of surfactant solution and two pore volumes of groundwater alone) were flushed to remove diesel from the site. The effluent TPH (total petroleum hydrocarbon) concentration with surfactant solution flushing increased to 1761 mg/L, which was over 200 times higher than the average concentration with only groundwater flushing. A total of 48 kg of TPH (about 88% of the initial TPH) was removed from the pilot site with five pore volumes of 2% sorbitan monooleate solution flushing; this total was more than 75 times the amount that was removed when flushing with water alone (less than 640 g). All of the extracted solution was treated by means of a chemical treatment process, which included the use of a dissolved air flotation system to lower the concentration of solution below 5mg/L and the treated solution was then disposed of in a nearby sewage drain.     

BTEX biodegradation by bacteria from effluents of petroleum refinery

Groundwater contamination with benzene, toluene, ethylbenzene and xylene (BTEX) has been increasing, thus requiring an urgent development of methodologies that are able to remove or minimize the damages these compounds can cause to the environment. The biodegradation process using microorganisms has been regarded as an efficient technology to treat places contaminated with hydrocarbons, since they are able to biotransform and/or biodegrade target pollutants. To prove the efficiency of this process, besides chemical analysis, the use of biological assessments has been indicated. This work identified and selected BTEX-biodegrading microorganisms present in effluents from petroleum refinery, and evaluated the efficiency of microorganism biodegradation process for reducing genotoxic and mutagenic BTEX damage through two test-systems: Allium cepa and hepatoma tissue culture (HTC) cells. Five different non-biodegraded BTEX concentrations were evaluated in relation to biodegraded concentrations. The biodegradation process was performed in a BOD Trak Apparatus (HACH) for 20 days, using microorganisms pre-selected through enrichment. Although the biodegradation usually occurs by a consortium of different microorganisms, the consortium in this study was composed exclusively of five bacteria species and the bacteria Pseudomonas putida was held responsible for the BTEX biodegradation. The chemical analyses showed that BTEX was reduced in the biodegraded concentrations. The results obtained with genotoxicity assays, carried out with both A. cepa and HTC cells, showed that the biodegradation process was able to decrease the genotoxic damages of BTEX. By mutagenic tests, we observed a decrease in damage only to the A. cepa organism. Although no decrease in mutagenicity was observed for HTC cells, no increase of this effect after the biodegradation process was observed either. The application of pre-selected bacteria in biodegradation processes can represent a reliable and effective tool in the treatment of water contaminated with BTEX mixture. Therefore, the raw petroleum refinery effluent might be a source of hydrocarbon-biodegrading microorganisms.     

Sulfolane biodegradation potential in aquifer sediments at sour natural gas plant sites

Sulfolane is used in the treatment of sour natural gas. It is a highly water soluble compound that has been introduced into soils and groundwaters at a number of sour gas processing plant sites. Aquifer sediments from contaminated locations at three sites in western Canada were assessed for microbial activity and their ability to degrade sulfolane under aerobic and five anaerobic (nitrate-, Mn(IV)-, Fe(III)-, sulfate- and CO₂-reducing) conditions. The microcosms were supplemented with 200 mg/L sulfolane and adequate supplies of N, P, and the appropriate terminal electron acceptor. Microcosms containing contaminated aquifer sediments from each of the three sites were able to degrade sulfolane aerobically at 8°C and 28°C, and the biodegradation followed zero-order kinetics. The lag times before the onset of sulfolane biodegradation were shorter when sulfolane-contaminated sediments were used as inocula than when uncontaminated soils were used. No anaerobic sulfolane biodegradation was observed at 28°C, nor was sulfolane biodegradation observed at 8°C under Fe(III)-, sulfate- and CO₂-reducing conditions. At 8°C, anaerobic degradation of sulfolane coupled to Mn(IV) reduction was observed in microcosms from two sites, and degradation coupled to nitrate reduction was seen in a microcosm from one of the contaminated sites.     

Plant tolerance to diesel minimizes its impact on soil microbial characteristics during rhizoremediation of diesel-contaminated soils

Soil contamination due to petroleum-derived products is an important environmental problem. We assessed the impacts of diesel oil on plants (Trifolium repens and Lolium perenne) and soil microbial community characteristics within the context of the rhizoremediation of contaminated soils. For this purpose, a diesel fuel spill on a grassland soil was simulated under pot conditions at a dose of 12,000 mg diesel kg^− 1 DW soil. Thirty days after diesel addition, T. repens (white clover) and L. perenne (perennial ryegrass) were sown in the pots and grown under greenhouse conditions (temperature 25/18 °C day/night, relative humidity 60/80% day/night and a photosynthetic photon flux density of 400 μmol photon m^− 2 s^− 1) for 5 months. A parallel set of unplanted pots was also included. Concentrations of n-alkanes in soil were determined as an indicator of diesel degradation. Seedling germination, plant growth, maximal photochemical efficiency of photosystem II (F_v/F_m), pigment composition and lipophylic antioxidant content were determined to assess the impacts of diesel on the studied plants. Soil microbial community characteristics, such as enzyme and community-level physiological profiles, were also determined and used to calculate the soil quality index (SQI). The presence of plants had a stimulatory effect on soil microbial activity. L. perenne was far more tolerant to diesel contamination than T. repens. Diesel contamination affected soil microbial characteristics, although its impact was less pronounced in the rhizosphere of L. perenne. Rhizoremediation with T. repens and L. perenne resulted in a similar reduction of total n-alkanes concentration. However, values of the soil microbial parameters and the SQI showed that the more tolerant species (L. perenne) was able to better maintain its rhizosphere characteristics when growing in diesel-contaminated soil, suggesting a better soil health. We concluded that plant tolerance is of crucial importance for the recovery of soil health during rhizoremediation of contaminated soils.     

Chemolithotrophic denitrification with elemental sulfur for groundwater treatment

Denitrification for the treatment of nitrates in wastewater typically relies on organic electron donating substrates. However, for groundwater treatment, inorganic compounds such as elemental sulfur (S0) are being considered as alternative electron donors in order to overcome concerns that residual organics can cause biofouling. In this study, a packed-bed bioreactor supplied with S0:limestone granules (1:1, v/v) was started up utilizing a chemolithotrophic denitrifying enrichment culture in the form of biofilm granules that was pre-cultivated on thiosulfate. The granular enrichment culture enabled a rapid start-up of the bioreactor. A nearly complete removal of nitrate (7.3 mM) was NO3- attained by the bioreactor at nitrate loading rates of up to 21.6 mmol/(L(reactor)d). With lower influent concentrations (1.3 mM nitrate) comparable to those found in contaminated groundwater, high nitrate loads of 18.1 mmol/(L(reactor)d) were achieved with an average nitrate removal efficiency of 95.9%. The recovery of nitrogen as benign N2 gas was nearly stoichiometric. The concentration of undesirable products from S0-based denitrification such as nitrite and sulfide were low. Comparison of bioreactor results with batch kinetic studies revealed that denitrification rates were dependent on the surface area of the added S0. The surface area normalized denitrification rate was determined to be 26.4 mmol /(m2 S0 d).     

超滤膜/混凝生物反应器去除饮用水中有机物的效能

采用超滤膜/混凝生物反应器(UF-MCBR)处理模拟微污染源水,考察了对有机污染物的去除效能与机理.结果表明,当聚合氯化铝投加量为10mg/L时,UF-MCBR对DOC、UV254、TOC、CODMn、BDOC和AOC的去除率分别为44.0%、54.5%、49.0%、58.5%、72.8%和58.3%.UF-MCBR通过超滤、生物降解以及混凝三者之间的协同作用去除溶解性有机物,就DOC的去除而言,三种作用的贡献率分别为11.1%、6.2%和26.7%.UF-MCBR系统中的UF膜表面为污泥层所覆盖,该污泥层能有效强化UF膜对分子质量为300-6000u 有机物的截留,UF膜(连同污泥层)对僧水碱、憎水中性物、憎水酸、弱憎水酸和亲水性物质的截留率分别达到了37.0%、42.8%、52.7%、39.8%和19.0%.