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1.
The effects of naphthalene concentrations and soil constituents (sand, clay, organic matter) on biodegrading naphthalene-contaminated soils using an acclimated Flavobacterium sp. were examined in continuously stirred slurry bioreactors. Soils with and without organic matter (1%) and kaolinite clay (20%) were used. Studies showed that sorption of naphthalene can be represented by the Freundlich isotherm. Among the soils investigated, clayey soil retarded the biodegradation process the most due to its desorption characteristics. Increasing the naphthalene contamination level from 500 mg∕kg to 25,000 mg∕kg doubled the biodegradation time in the slurry reactor with a soil loading of 10 g∕L. A pseudo first-order kinetic was observed for naphthalene biodegradation, and the biodegradation constant was determined to be 0.20 mg∕L∕h. A kinetic model was developed to simulate the biodegradation of naphthalene in a continuously stirred batch slurry reactor and to understand the role of solubilization (solid phase naphthalene), desorption, and biodegradation in the removal of naphthalene. Predictions using the numerical model agreed with the experimental data.  相似文献   

2.
The biodegradability of two surfactants, Triton X-100 and Rhamnolipid, was tested under aerobic, nitrate reducing, sulfate reducing, and anaerobic conditions in a respirometer. The results indicated that from a biodegradation standpoint, Rhamnolipid is superior to Triton X-100 since it is biodegradable under all conditions, whereas the Triton X-100 is partially biodegradable under aerobic conditions and nonbiodegradable under anaerobic, nitrate reducing, and sulfate reducing conditions.  相似文献   

3.
Two-dimensional separation of erythrocyte membrane proteins   总被引:1,自引:0,他引:1  
1). Erythrocyte membrane proteins eluted with Triton X-100 or dilute EDTA have been separated two-dimensionally by isoelectric focusing in polyacrylamide gels containing 1 percent Triton X-100 plus 8 M urea, followed by electrophoresis using sodium dodecyl sulfate. Characteristic patterns, consistent among 40 healthy donors, were obtained. 2. The resulting patterns contain at least 30 components. The "spectrin" components (sodium dodecyl sulfate Bands 1 and 2) focus in the same pH range. Other membrane components giving single bands in sodium dodecyl sulfate electrophoresis appear to be heterogeneous. 3. Triton X-100, but not EDTA, extracts the principal membrane glycoproteins and the major "intrinsic" protein. Otherwise, proteins preferentially eluted by EDTA extract poorly with Triton X-100 and vice versa. 4. Membrane glycoproteins migrate anodally during electrofocusing and can be purified in a simple, one-step procedure.  相似文献   

4.
The application of surfactants for the bioremediation of sites contaminated with polycyclic aromatic hydrocarbons has been widely reported, because they are known to increase PAH solubility and desorption, thereby enhancing their bioavailability to biofilm microorganisms. The effects of a nonionic surfactant on the biodegradation of PAHs in porous media, as well as the fate of the surfactant, were investigated in this study. Column experiments in the presence of the surfactant showed that the degradation of the two-ring PAH alone was not significantly affected, but that there was a small enhancement of three- and four-ring PAH degradation when they were present as sole substrates and when using Triton X-100. This was due to the higher solubility of the PAHs in the presence of the surfactant. Biofilm seemed to respond well to binary mixtures of phenanthrene–naphthalene and pyrene–naphthalene, with removals of 45.5 and 24.1%, respectively, in the presence of the surfactant; however, higher biodegradations were always achieved by having just PAH mixtures without the surfactant, indicating the importance of cometabolic mechanisms over improved solubilization of PAHs. Optical sections taken using a confocal laser scanning microscope allowed observation of a heterogeneous web-like matrix of biofilm, with diverse biological aggregate structures.  相似文献   

5.
Activated sludge sequencing batch reactors were used to study BTX biodegradation under anoxic (denitrifying), microaerobic, and aerobic conditions. Toluene and m-xylene were biodegraded under denitrifying conditions, and the loss of these compounds correlated with the activity of reducing enzymes that were capable of oxidizing methyl viologen. Although benzene, p-, and o-xylene were recalcitrant under anoxic treatment, all three were biodegraded under microaerobic [<0.2 mg∕L dissolved oxygen (DO)] and nitrate or nitrite (NOx)-supplemented microaerobic conditions. Methyl viologen oxidation potential decreased under all microaerobic conditions while catechol 1,2-dioxygenase (C12O) and catechol 2,3-dioxygenase (C23O) were induced, indicating that the aromatic hydrocarbons were metabolized by aerobic pathways, even in the presence of NOx and in the absence of measurable DO. The degree of C12O and C23O expression under microaerobic conditions was comparable to levels found under aerobic (DO > 4 mg∕L) conditions. Benzene, p-, and o-xylene were biodegraded twice as fast under NOx-supplemented compared to NOx-free microaerobic conditions, and specific biodegradation rates under aerobic and NOx-supplemented microaerobic conditions were comparable. Oxidation reduction potential successfully differentiated between the various electron acceptor conditions and proved to be a sensitive indicator.  相似文献   

6.
A microbial surfactant (biosurfactant) was investigated for its potential to enhance bioavailability and, hence, the biodegradation of octadecane. The rhamnolipid biosurfactant used in this study was extracted from culture supernatants after growth of Pseudomonas aeruginosa ATCC 9027 in phosphate-limited proteose peptone-glucose-ammonium salts medium. Dispersion of octadecane in aqueous solutions was dramatically enhanced by 300 mg of the rhamnolipid biosurfactant per liter, increasing by a factor of more than 4 orders of magnitude, from 0.009 to > 250 mg/liter. The relative enhancement of octadecane dispersion was much greater at low rhamnolipid concentrations than at high concentrations. Rhamnolipid-enhanced octadecane dispersion was found to be dependent on pH and shaking speed. Biodegradation experiments done with an initial octadecane concentration of 1,500 mg/liter showed that 20% of the octadecane was mineralized in 84 h in the presence of 300 mg of rhamnolipid per liter, compared with only 5% octadecane mineralization when no surfactant was present. These results indicate that rhamnolipids may have potential for facilitating the bioremediation of sites contaminated with hydrocarbons having limited water solubility.  相似文献   

7.
Solid waste produced from construction and demolition (C&D) activities is typically disposed of in unlined landfills. Knowledge of C&D debris landfill leachate is limited in comparison to other types of wastes. A laboratory study was performed to examine leachate resulting from simulated rainfall infiltrating a mixed C&D waste stream consisting of common construction materials (e.g., concrete, wood, drywall). Lysimeters (leaching columns) filled with the mixed C&D waste were operated under flooded and unsaturated conditions. Leachate constituent concentrations in the leachate from specific waste components were also examined. Leachate samples were collected and analyzed for a number of conventional water quality parameters including pH, alkalinity, total organic carbon, total dissolved solids, and sulfate. In experiments with the mixed C&D waste, high concentrations of total dissolved solids (TDS) and sulfate were detected in the leachate. C&D leachates produced as a result of unsaturated conditions exhibited TDS concentrations in the range of 570–2,200 mg∕L. The major contributor to the TDS was sulfate, which ranged in concentration between 280 and 930 mg∕L. The concentrations of sulfate in the leachate exceeded the sulfate secondary drinking water standard of 250 mg∕L. The leachate produced from lysimeters exposed to conditions of constant flooding possessed a greater concentration of dissolved constituents than leachate from the unsaturated lysimeters. The sulfate concentration ranged from 950 to 1100 mg∕L. In both scenarios, the primary ions contributing to the dissolved solids were sulfate and calcium, both a result of gypsum drywall. The flooded lysimeters remained at a constant pH of 11 throughout the experiment, whereas the pH dropped to neutral conditions (pH 6–7) in the unsaturated columns after 1 month of the leaching experiment. The high pH of leachate from the flooded columns was attributed to concrete. Based on leaching tests on individual waste components, wood and cardboard were the primary materials contributing to dissolved organic carbon. The organic carbon concentrations in the leachate were generally lower than typical municipal waste leachate. The biological conversion of sulfate to sulfide was evident in many columns, and was most pronounced in the unsaturated, mixed-waste columns.  相似文献   

8.
Pseudomonas sp. D8 strain, which has the potential to utilize toluene as a sole carbon source, was isolated. At a concentration of 100 mg/l, this strain was found to efficiently degrade toluene and benzene (both individually and in mixture) in culture medium at 30 degrees C and pH7. Following a two-hour lag phase, complete biodegradation of 100 mg/l toluene or benzene occurred within 6 to 8 hours. The addition of nitrate, phosphate, or sulfate at various concentrations were found to have significant influence on both toluene and benzene degradation. In addition, results show that the D8 strain has the ability to degrade monochlorophenols, nitrophenols, and phenol, but not aliphatic compounds. Inoculation of groundwater samples containing 100 mg/l toluene or benzene with Pseudmonas sp. D8 resulted in rapid degradation within 24-33 hours.  相似文献   

9.
The effects of exposure of Culex pipiens larvae to sublethel concentrations of larvicides on uptake, development of Wuchereria bancrofti, survival rate and reproduction of filaria-infected mosquitoes were investigated. Fourth instar larvae of Cx. pipiens were exposed to LC40 of the surfactant Triton X-100, the insect growth regulator DPX alone or combined with LC10 of the surfactant and permethrin alone or combined with LC10 of the surfactant. Adults that survived insecticide treatments and controls were infected by allowing them to feed on microfilaremic volunteers. Significant reduction in the uptake of microfilaria was observed in groups treated with Triton X-100 alone or combined either with permethrin or DPX when compared to control. The overall infection and infective rates were significantly reduced in mosquitoes treated with Triton X-100 either alone or combined with permethrin. Treatment with Triton X-100 and DPX prolonged the extrinsic incubation period (EIP) and retarded the development of filarial larvae, while permethrin either alone or combined with Triton X-100 and DPX combined with Triton X-100 shortened the EIP. All larvicides reduced the number of infective larvae (L3)/mosquito and induced deformities among he different parasite stages, especially in mosquitoes treated with combination of permethrin and Triton X-100 or mixture of DPX and Triton X-100 where 36% and 54.9% respectively of L3S were deformed. In treated mosquitoes, a low percentage of L3S was detected in the head and proboscis region while the majority was trapped in the thoracic region. The survival rates of mosquitoes were reduced in cases treated with permethrin, DPX and Triton X-100 while treatment with mixture of DPX and Triton X-100 induced higher rate of mortalities when compared to control. Egg production of filaria- infected Cx. pipiens was significantly reduced in mosquitoes treated with DPX and Triton X-100. It was observed that the addition of Triton X-100 to DPX or to permethrin significantly reduced egg production. The results suggest that sublethal concentrations of larvicides especially Triton X-100 applied to 4th instar larvae of Cx. pipiens could effectively interfere with the development of W. bancrofti in Cx. pipiens and reduced the survival rate and fecundity of the vector.  相似文献   

10.
A novel beta-glucosidase, which is inducible and capable of catalyzing the hydrolysis of sennosides, was purified from Bifidobacterium sp. strain SEN with Triton X-100 solubilization and DEAE-cellulose column chromatography, by which hydrolytic activities toward sennoside B, 4-methylumbelliferyl beta-glucoside (MUG), and p-nitrophenyl beta-glucoside (pNPG) were obtained together in the same eluted fractions. The activity was stable against detergents such as sodium dodecyl sulfate (SDS) and Triton X-100, but was denatured by SDS and beta-mercaptoethanal when heated. The final preparation was shown to be nearly homogeneous on SDS-polyacrylamide gel electrophoresis (PAGE) either after the enzyme was denatured or when it was not denatured. In the non-denaturing SDS-PAGE, a single protein band hydrolyzed MUG on the gel. In the denaturing SDS-PAGE, the subunit mass of the enzyme was estimated to be 110 kDa. The enzyme was optimally active at pH 6.0 for hydrolysis of sennoside B and MUG. Km values for sennoside B and MUG are 0.94 and 0.53 mM, respectively. The enzyme also catalyzed the hydrolysis of pNPG, amygdalin, geniposide and salicin. It was less active against methyl beta-glucoside and incapable of hydrolyzing cellobiose. The beta-glucosidase activity was inhibited by deoxynojirimycin and p-chloromercuribenzenesulfonic acid, but was less susceptible to several metals (FeSO4, ZnCl2, and CuSO4), and 5,5'-dithio-bis(2-nitrobenzoic acid).  相似文献   

11.
Phospholipase D [phosphatidylcholine cholinehydrolase, EC 3.1.4.4] excreted from Streptomyces chromofuscus was purified from the culture supernatant by precipitation with acetone and column chromatographies on palmitoylated gauze (Pal-G), DEAE-cellulose, and Sephadex G-150 with an overall recovery of 46% and 1000-fold increase in specific activity. The purified enzyme preparation showed a single band on sodium dodecyl sulfate (SDS) polyacrylamide disc gel electrophoresis. The enzyme had a molecular weight of about 50,000 by gel filtration on Sephadex G-150 or about 57,000 by SDS-polyacrylamide disc gel electrophoresis and an isoelectric point (pI) of pH 5.1 on isoelectric focusing. The enzyme hydrolyses lecithin, lysolecithin, sphingomyelin, and cephalin; the relative reaction velocities and Km's for choline-phospholipids were 87% and 1.43 mM for lecithin, 100% and 1.67 mM for lysolecithin, and 22% and 0.56 mM for sphingomyelin. The enzymatic reaction was optimal at pH 8, and its velocity was appreciably increased by either detergent (Triton X-100, deoxycholate), Ca2+ or both detergent and Ca2+. Diethyl ether stimulated the enzymatic activity by 30%; SDS and EDTA inhibited the activity. Bovine serum albumin, Triton X-100, and lipids (lecithin, lysolecithin, phosphatidic acid, lysophosphatidic acid, palmitic acid, and oleic acid) inhibited adsorption of the purified enzyme onto palmitoyl cellulose (Pal-C) and affected both the enzyme activity and stability: albumin and Triton X-100 increased the activity and enhanced the heat-stability; lysophospholipids decreased the activity but other lipids increased the activity; all the lipids lowered the heat-stability. The enzyme adsorbed on Pal-C was active, although its activity was about one-ninth of that of free enzyme, and was protected from heat-inactivation. Thus this enzyme appears to possess a hydrophobic site distinct from its catalytic site and to be adsorbed onto Pal-C through the hydrophobic site. Albumin, Triton X-100, and lipids seem to bind to the hydrophobic site and to have an appreciable effect on the enzyme activity and stability.  相似文献   

12.
Immobilized-Cell Membrane Bioreactor for High-Strength Phenol Wastewater   总被引:1,自引:0,他引:1  
An immobilized-cell membrane bioreactor was fabricated to investigate degradation of phenol at high concentrations using Pseudomonas putida American Type Culture Collection 49451. In the case of suspension cultures, P. putida utilized phenol at concentrations below 1,000 mg∕L, but experienced substrate inhibition at higher concentrations. On the other hand, cells immobilized in 25% by weight polysulfone fibers degraded phenol at concentrations above 1,000 mg∕L. At an initial phenol concentration of 1,200 mg∕L, phenol was fully degraded within 95 h in the immobilized system, whereas no cell growth and phenol degradation were observed in the free suspension system at 1,000 mg∕L phenol. In the immobilized system, it was observed that cells diffused from the membranes when phenol concentrations reached noninhibitory levels in a few experiments. In such cases, the time taken for complete degradation was shorter with cell diffusion because suspension cells were responsible for the rapid phenol degradation. Further biodegradation studies at phenol concentrations of 2,000 and 3,500 mg∕L were also performed to evaluate the effectiveness of cell immobilization for delaying the effects of substrate inhibition. Phenol could be completely degraded at both high concentrations.  相似文献   

13.
Entry of Listeria monocytogenes into nonphagocytic cells requires the inlAB gene products. InlA and InlB are bacterial cell wall-associated polypeptides that can be released by sodium dodecyl sulfate treatment. By applying more gentle extraction methods, we have purified InlB in its native form. Treatment of bacteria with various nondenaturating agents including mutanolysin, thiol reagents, sodium chloride, and detergents like Triton X-100 or 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate did not release substantial amounts of InlB from the bacterial cell wall. Instead, InlB was nearly quantitatively extracted in a solubilized form by treatment of bacteria with 1 M Tris-Cl or other protonated amines at pH 7.5. However, the reduced solubility of the extracted InlB in low-salt buffers hampered further biochemical purification. A panel of monoclonal antibodies against listerial Tris-Cl extracts containing InlB was therefore produced to generate reagents for use in affinity chromatography. One of the monoclonal antibodies enabled purification of the InlB protein to homogeneity with relatively high yields. When added externally, purified InlB associated with the surface of noninvasive bacteria such as Listeria innocua or an L. monocytogenes inlB2 mutant, where it promoted entry of these strains into Vero cells >300- and 17-fold, respectively. This effect was even more dramatic for HeLa cells, where the observed invasion was increased about 9,000- and 4,000-fold, respectively. The availability of purified native, invasion-competent InlB will allow analysis of the molecular basis of InlB-mediated entry into tissue culture cell lines in greater detail.  相似文献   

14.
The micellar properties of mixtures of GM1 ganglioside and the non-ionic amphiphile Triton X-100 in 25 MM Na phosphate-5 mM di Na EDTA buffer (pH = 7.0) were investigated by quasielastic light scattering in a wide range of Triton/GM1 molar ratios and in the temperature range 15-37 degrees C. These measurements: (a) provided evidence for the formation of mixed micelles; (b) allowed the determination of such parameters as the molecular weight and the hydrodynamic radius of the mixed micelles; (c) showed the occurrence of statistical aggregates of micelles with increasing temperature and micelle concentration. Galactose oxidase was chosen for studying the relation between enzyme activity and micellar properties. The action of the enzyme on GM1 was found to be strongly dependent on the micellar structure. In particular: (a) galactose oxidase acted very poorly on homogeneous GM1 micelles, while affecting mixed GM1/Triton X-100 micelles; (b) at fixed GM1 concentration the oxidation rate increased by enhancing Triton X-100 concentration and followed a biphasic kinetics with a break at a certain Triton X-100 concentration; (c) the formation of statistical micelle aggregates was followed by inhibition of the enzyme activity.  相似文献   

15.
Competitive Substrate Biodegradation during Surfactant-Enhanced Remediation   总被引:2,自引:0,他引:2  
The impact of synthetic surfactants on the aqueous phase biodegradation of benzene, toluene, and p-xylene (BTpX) was studied using two anionic surfactants, sodium dodecyl sulfate (SDS) and sodium dodecyl benzene sulfonate (SDBS), and two nonionic surfactants, POE(20) sorbitan monooleate (T-maz-80) and octylphenolpoly(ethyleneoxy) ethanol (CA-620). Batch biodegradation experiments were performed to evaluate surfactant biodegradability using two different microbial cultures. Of the four surfactants used in this study, SDS and T-maz-80 were readily degraded by a microbial consortium obtained from an activated sludge treatment system, whereas only SDS was degraded by a microbial culture that was acclimated to BTpX. Biodegradation kinetic parameters associated with SDS and T-maz-80 degradation by the activated sludge consortium were estimated using respirometric data in conjunction with a nonlinear parameter estimation technique as μmax = 0.93 h?1, Ks = 96.18 mg∕L and μmax = 0.41 h?1, Ks = 31.92 mg∕L, respectively. When both BTpX and surfactant were present in the reactor along with BTpX-acclimated microorganisms, two distinct biodegradation patterns were seen. SDS was preferentially utilized inhibiting hydrocarbon biodegradation, whereas the other three surfactants had no impact on BTpX biodegradation. None of the four surfactants were toxic to the microbial cultures used in this study. Readily biodegradable surfactants are not very effective for subsurface remediation applications as they are rapidly consumed, and also because of their potential inhibitory effects on intrinsic hydrocarbon biodegradation. This greatly increases treatment costs as surfactant recovery and reuse are adversely affected.  相似文献   

16.
Removal of Lead from Contaminated Water and Clay Soil Using a Biosurfactant   总被引:2,自引:0,他引:2  
Lead removal from water and contaminated soils was investigated using biosurfactant, anionic, and nonionic surfactants in continuously stirred batch reactors. Lead-contaminated water up to 100?mg/L and clay soil up to 3,000?mg/kg were used in this investigation. The surfactant concentration up to 10 critical micelle concentration was used. The speciation of lead into the micelles was quantified and the lead removal efficiency depended on the level of contamination, surfactant type, and concentration. Of the surfactants used, biosurfactant (produced from used vegetable oil) had the best removal efficiency (75%) at a lead contamination of 100?mg/L in water at pH of over 12. The Fourier-transformed infrared spectroscopy study showed that the carboxyl group in the biosurfactant was effective in removing the lead from the solution. Langmuir and Freundlich relationships were used to represent the micelle partitioning of lead in the surfactant solutions. Desorption of lead from contaminated kaolinite clay was represented using linear isotherms. The biosurfactant solution had a higher micelle partitioning for the lead from contaminated water and desorbing the lead from the contaminated soil compared to the other chemical surfactants.  相似文献   

17.
Triton X-100 inhibits the NADH oxidase and rotenone-sensitive NADH-Q1 reductase activities of bovine heart submitochondrial particles (SMP) with an apparent Ki of 1x10-5 M (pH 8.0, 25 degrees C). The NADH-hexammineruthenium reductase, succinate oxidase, and the respiratory control ratio with succinate as the substrate in tightly coupled SMP are not affected at the inhibitor concentrations below 0.15 mM. The succinate-supported aerobic reverse electron transfer is less sensitive to the inhibitor (Ki=5x10-5 M) than NADH oxidase. Similar to rotenone, limited concentrations of Triton X-100 increase the steady-state level of NAD+ reduction when the nucleotide is added to tightly coupled SMP oxidizing succinate aerobically. Also similar to rotenone, Triton X-100 partially protects Complex I against the thermally induced deactivation and partially activates the thermally deactivated enzyme. The rate of the NADH oxidase inhibition by rotenone is drastically decreased in the presence of Triton X-100 which indicates a competition between these two inhibitors for a common specific binding site. In contrast to rotenone, the inhibitory effect of Triton X-100 is instantly reversed upon dilution of the reaction mixture. The NADH-Q1 reductase activity of SMP is inhibited non-competitively by added Q1 whereas a simple competition between Q1 and the inhibitor is seen for isolated Complex I. The results obtained show that Triton X-100 is a specific inhibitor of the ubiquinone reduction by Complex I and are in accord with our previous findings which suggest that different reaction pathways operate in the forward and reverse electron transfer at this segment of the mammalian respiratory chain.  相似文献   

18.
To determine if membrane association is an intrinsic property of the influenza virus matrix protein (M1) it was expressed from cDNA in living cells in the absence of other influenza virus proteins. By using a membrane fractionation scheme the M1 protein was found to associate with membranes in a time-dependent manner (0 time = 45% total; after a 3-hr chase period = 68% total M1 protein). Coexpression of the integral membrane proteins HA+NA+M2 did not significantly increase the association of the M1 protein with cellular membranes, indicating that putative interactions of the M1 protein and the cytoplasmic tails of the integral membranes cannot be detected by this assay. Biochemical treatments of the M1 protein associated with membranes with alkali, high salt conditions, or Triton X-114 yielded data that challenge the normal criteria for integral membrane proteins or peripheral membrane proteins. Examination of the solubility of the M1 protein in influenza virus-infected cells to Triton X-100 extraction indicated it became increasingly insoluble with time, but the M1 protein could be solubilized in Triton X-100 containing 1 M NaCl, suggesting an association of the M1 protein with the cytoskeleton. However, when the M1 protein was expressed from cDNA, it did not become insoluble to Triton X-100 extraction, suggesting an interaction of the M1 protein unique to the influenza virus-infected cell.  相似文献   

19.
The numbers of Triton X-100 and sodium dodecyl sulfate molecules required to form respective pores were estimated from the relationship between the detergent concentrations and the rates of fast and slow hemolysis components. It has been found that the slow hemolysis component evoked by Triton X-100 is related to the existence of two different pores. It is shown that the fast hemolysis component induced by sodium dodecyl sulfate is associated with the modification of phosphatidylcholine which determines the break in the Arrhenius plots of the hemolysis rate within the range of 20 degrees C. The shape of hemolysis kinetic curves and the dependence of hemolytic parameters on the detergent concentration and temperature are discussed based on the concept of hemolysis caused by the formation of pores in various membrane lipid regions and by releasing vesicles from erythrocytes.  相似文献   

20.
在实验室保证厌氧条件下,通过恒温摇床试验,研究焦化废水中芳烃类物质苯胺、萘和茚的降解去除规律,同时对这三种芳烃类有机物在水相和泥相中的含量进行了测定。研究结果表明:苯胺的厌氧去除主要通过生物降解,但微生物需要一个适应过程;萘和茚的厌氧去除途径是吸附在污泥上进行降解,茶经过48h可降解完全,茚的吸附属物理吸附,是可逆的。  相似文献   

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