Comparison of soluble microbial products released from activated sludge and aerobic granular sludge systems in the presence of toxic 2,4-dichlorophenol

Bioprocess and Biosystems Engineering - The objective of this study was to compare the release of soluble microbial products (SMP) from activated sludge (AS) and aerobic granular sludge (AGS) in...     

Effect of sludge age on performance of an activated sludge unit treating 2,4 dichlorophenol-containing synthetic wastewater

Wastewaters containing chlorophenol compounds are difficult to treat by biological means because of toxic effects of chlorophenols on microorganisms. Synthetic wastewater containing 2,4 dichlorophenol (DCP) was biologically treated in an activated sludge unit at different sludge ages varying between 5 and 30 days while the feed COD, DCP contents and hydraulic residence time (HRT) were constant. Effects of sludge age on COD, DCP and toxicity removals were investigated. Increases in sludge age caused significant increases in biomass concentration in the aeration tank, which resulted in increases in percent COD, DCP and toxicity removals. COD removal increased from 58 to 90%, while DCP and toxicity removals increased from 15 to 100% and from 38 to 100%, respectively, when the sludge age was raised from 5 to 30 days. Resazurin method based on dehydrogenase activity was used for assessment of the feed and effluent wastewater toxicity. Sludge volume index (SVI) decreased with increasing sludge age indicating improved settling characteristics of the sludge at high sludge ages. Operation at a sludge age of 25 days resulted in more than 90% COD and nearly 100% DCP and toxicity removal with an SVI value of 108 ml g⁻¹ under the experimental conditions tested.     

Influence of 2,4-dinitrophenol on the characteristics of activated sludge in batch reactors

The response of activated sludge characteristics to the presence of 2,4-dinitrophenol (dNP) in batch cultures was investigated in this study. The sludge yield slightly decreased with an increase in dNP concentration. At 10 mg l(-1), or lower, dNP significantly reduced sludge yield and relative specific growth rates (mu/mu0), but didn't substantially affect its relative specific chemical oxygen demand removal rate (q/q0). Presence of dNP at 1-20 mg l(-1) increased the specific oxygen uptake rate of activated sludge, and slightly changed its hydrophobicity. An analysis on inhibition indicated that the reduction in sludge yield in the presence of dNP was mainly attributed to the significant decreased sludge growth, rather than the reduced substrate degradation.     

Effects of 2,4-D and its metabolite 2,4-dichlorophenol on antioxidant enzymes and level of glutathione in human erythrocytes

The effects of in vitro exposure of human erythrocytes to different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and its metabolite 2,4-dichlorophenol (2,4-DCP) were studied. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the level of reduced glutathione (GSH) were determined. The activity of erythrocyte superoxide dismutase SOD decreased with increasing dose of 2,4-D and 2,4-DCP, while glutathione peroxidase activity increased. 2,4-D (500 ppm) decreased the level of reduced glutathione in erythrocytes by 18% and 2,4-DCP (250 ppm) by 32%, respectively, in comparison with the controls. These results lead to the conclusion that in vitro administration of herbicide-2,4-D and its metabolite 2,4-DCP causes a decrease in the level of reduced glutathione in erythrocytes and significant changes in antioxidant enzyme activities. Comparison of the toxicity of 2,4-D and 2,4-DCP revealed that the most prominent changes occurred in human erythrocytes incubated with 2,4-DCP.     

Modeling the yield of activated sludge in the presence of 2,4-dinitrophenol

In this work, a model was developed to evaluate the yield of activated sludge in the presence of 2,4-dinitrophenol (dNP). Experimental results show that the presence of dNP substantially reduced the sludge yield at all solid retention times tested, but that it had no negative effect on the bioactivity and settleability of activated sludge. With the established model, the maximum observed sludge yield (Y^′_max) and specific endogenous respiration rate (k_d) for the sludge without dNP dose were estimated as 0.79 g-MLSS/g-COD and 0.042 d⁻¹, respectively, whereas for the sludge dosed with dNP the corresponding values were calculated as 0.56 g-MLSS/g-COD and 0.056 d⁻¹, respectively. In the presence of dNP at 5 mg/l, the energy uncoupling coefficient was calculated as 0.29.     

Preparation of anti-2,4-dichlorophenol and 2,4-dichlorophenoxyacetic acid monoclonal antibodies

The ratios of hapten and bovine serum albumin (BSA) in an antigen conjugate were determined by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Hybridomas secreting monoclonal antibodies against 2,4-dichlorophenoxyacetic acid (2,4-D) were produced by fusing 2,4-D-BSA conjugate-immunized splenocytes with a HAT-sensitive mouse myeloma cell line, P3-X63-Ag8-653. A substantial cross-reaction was observed for 2,4-dichlorophenol (2,4-DP) when compared with that observed for 2,4-D. The full measurement range for this assay is 0.2–3 μg ml⁻¹ for 2,4-DP. On the other hand, the range for 2,4-D is between 1 and 20 μg ml⁻¹. This revised version was published online in July 2006 with corrections to the Cover Date.     

Behavior of 2,4-dichlorophenoxyacetic acid degradation and nitrogen conversion by an activated sludge

Summary Degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) was examined together with nitrogen conversion by using an activated sludge acclimated to artificial sewage containing 2,4-D and urea-N, which were the sole carbon and nitrogen sources, respevtively. Ammonification of urea and nitrification of ammonia proceeded concurrently with 2,4-D degradation by the acclimated activated sludge.     

Purification and properties of a plasmid-encoded 2,4-dichlorophenol hydroxylase

2,4-Dichlorophenol hydroxylase, an enzyme involved in the bacterial degradation of the herbicide 2,4-dichlorophenoxyacetate (2,4-D) was purified from two bacterial strains that harbored the same 2,4-D plasmid, pJP4. The purified enzymes (Mr 224 000) from the two transconjugants were indistinguishable; they contained FAD and were composed of non-identical subunits, Mr 67 000 and 45 000, respectively. Various substituted phenols were hydroxylated, using either NADH or NADPH. The amino acid composition of the native enzyme was determined.     

Sequential anaerobic degradation of 2,4-dichlorophenol in freshwater sediments

2,4-Dichlorophenol (2,4-DCP) was anaerobically degraded in freshwater lake sediments. From observed intermediates in incubated sediment samples and from enrichment cultures, the following sequence of transformations was postulated. 2,4-DCP is dechlorinated to 4-chlorophenol (4-CP), 4-CP is dechlorinated to phenol, phenol is carboxylated to benzoate, and benzoate is degraded via acetate to methane and CO2; at least five different organisms are involved sequentially. The rate-limiting step was the transformation of 4-CP to phenol. Sediment-free enrichment cultures were obtained which catalyzed only the dechlorination of 2,4-DCP, the carboxylation of phenol, and the degradation of benzoate, respectively. Whereas the dechlorination of 2,4-DCP was not inhibited by H2, the dechlorination of 4-CP, and the transformation of phenol and benzoate were. Low concentrations of 4-CP inhibited phenol and benzoate degradation. Transformation rates and maximum concentrations allowing degradation were determined in both freshly collected sediments and in adapted samples: at 31 degrees C, which was the optimal temperature for the dechlorination, the average adaptation time for 2,4-DCP, 4-CP, phenol, and benzoate transformations were 7, 37, 11 and 2 days, respectively. The maximal observed transformation rates for these compounds in acclimated sediments were 300, 78, 2, 130, and 2,080 micromol/liter(-1)/day(-1), respectively. The highest concentrations which still allowed the transformation of the compound in acclimated sediments were 3.1 m/M 2,4-DCP, 3.1 mM 4-CP, 13 mM phenol, and greater than 52 mM benzoate. The corresponding values were lower for sediments which had not been adapted for the transformation steps.     

Biodegradation kinetics of 2,4-dichlorophenol by acclimated mixed cultures

The biodegradation kinetics of 2,4-dichlorophenol (2,4-DCP) by culture (Culture M) acclimated to mixture of 4-chlorophenol (4-CP) and 2,4-DCP and the culture (Culture 4) acclimated to 4-CP only were investigated in aerobic batch reactors. Also, pure strains isolated from mixed cultures were searched for their ability towards the biodegradation of 2,4-DCP. Culture 4 was able to completely degrade 2,4-DCP up to 80 mg/L within 30 h and removal efficiency dropped to 21% upon increasing initial concentration to 108.8 mg/L. When the Culture M was used, complete degradation of 2,4-DCP in the range of 12.5-104.4 mg/L was attained. A linear relationship between time required for complete degradation and initial 2,4-DCP concentrations was observed for both mixed cultures. It was observed that the Haldane equation can be used to predict specific degradation rate (SDR) (R(2)>0.99) as a function of initial 2,4-DCP concentrations and it adequately describes 2,4-DCP concentration profiles. Both of the mixed cultures settled well, which is important to maintain good removal efficiency for longer periods of time for real full-scale applications. Although the pure strains isolated from mixed cultures were found to have higher SDR of 2,4-DCP compared to mixed cultures, they did not settle well under quiescent conditions.     

Effects of cationic surfactants on settling properties of sewage activated sludge

Various alkyltrimethylammonium bromides ATABs, R = C₁ to C₆ were added to a mixed liquor of sewage activated sludge and the effects on settling properties were investigated. The 30-min settled sludge volume decreased from 28% to 24, 20, 17, and 15% on addition of 1 mM C₁₀, C₁₂, C₁₄, and C₁₆ compounds to mixed liquor (sludge concentration, 1.28 g/l), respectively. With increasing alkyl chain length, the amounts of ATABs adsorbed by the sludge, and metal ions released from the sludge, increased. However, a quantitative correlation was not observed between the two. In addition, the adsorption of ATABs resulted in a small decrease in sludge wet weight. However, such a change was not observed on addition of ATABs bearing short alkyl chains. A good correlation was observed between the amounts of ATABs adsorbed and their organic solvent/water partition coefficients. These results indicated that the effectiveness of ATABs as sludge conditioners depends on their hydrophobicity.     

Effects of synthetic polymer on the filamentous bacteria in activated sludge

Filamentous bulking is one of the solid-liquid separation problems always seen in activated sludge process. The addition of synthetic polymer is always one of the popular ways for the treatment plant operator to immediately solve the poor sludge settling problem. Therefore, it may be interesting to understand the effects of synthetic polymer on the filamentous bacteria in activated sludge. In this study, synthetic polymer was applied to a lab-scale wastewater treatment system with the filamentous bulking problem. The population structure of filamentous bacteria and sludge characteristics were investigated under different conditions. When synthetic polymer was added into the system, it was found that poor sludge settleability caused by filamentous bulking was temporarily solved and filamentous branches growing outside the flocs were damaged or inhibited. However, filamentous growth was still observed inside the flocs. After the addition of polymer was halted, filamentous branches extended out of the flocs immediately. Very serious filamentous bulking occurred and sludge settleability became much worse than that occurring before the addition of polymer. And, it took several weeks for the system to return to normal operation.     

Studies on organic removal of 2,4-dichlorophenol wastewaters using a modified RBC

Phenolic compound wastes from a large number of industries big and small which are highly toxic and pose a direct threat to human and aquatic life are generally let into the rivers and coastal waters. 2,4-dichlorophenol is used in the manufacture of industrial and agricultural products such as pesticides, germicides, soil sterilants, seed disinfectants and antiseptics. A modified Rotating Biological Contactor (RBC) was used for the treatability studies of synthetic 2,4-di-chlorophenolic (2,4 CP) wastewaters. The RBC used was a four stage laboratory model and the discs were modified by attaching porous netlon sheets to enhance biofilm area and volume. Synthetic wastewaters were prepared with influent concentrations from 40 to 200?mg/l of 2,4 CP. Four hydraulic loads were used in the range of 0.024 to 0.065 m³.m^-2.d^-1 and the organic loads used were in the range of 2 to 13?g 2,4 CP.m^-2.d^-1. The RBC was operated at a speed of 12?rpm. Effect of hydraulic loadings and influent 2,4-dichlorophenol concentration on 2,4-dichlorophenol removal were discussed and showed maximum organic removal at hydraulic loads of 0.024 and 0.046?m³.m^-2.d^-1. Also, a correlation plot between 2,4 CP applied and 2,4 CP removed was presented. A mathematical model was proposed using regression analysis.     

Effects and model of alkaline waste activated sludge treatment

Due to lack of information about alkaline sludge treatment with high dose at ambient temperature, alkaline sludge treatment was investigated to know the effects of different doses (0-0.5 mol/L) of sodium hydroxide (NaOH) and calcium hydroxide [Ca(OH)2] at 0-40 degrees C. Results showed that NaOH was more suitable than Ca(OH)2 for sludge disintegration. For NaOH treatment, most efficient dose was about 0.05 mol/L (0.16 g/g dry solid), and 60-71% solubilization of organic matters was achieved in first 30 min during its treatment of 24h. The process is described by a transmutative power function model, and relation between reaction rate constant and temperature are according with Arrhenius equation. Low dose NaOH treatment (<0.2 mol/L) deteriorated sludge dewatering ability obviously, while the ability was recovered gradually with the increase of NaOH dose. For Ca(OH)2 treatment, the disintegrated floc fragments and soluble organic polymers can be re-flocculated with the help of calcium cations. Consequently sludge disintegration effect was counteracted and dewatering ability improved.     

Genotoxic effect of 2,4-dichlorophenoxy acetic acid and its metabolite 2,4-dichlorophenol in mouse.

The cytogenetic effect of 2,4-dichlorophenoxy acetic acid (2,4-D) and its metabolite 2,4-dichlorophenol (2,4-DCP) was studied in bone-marrow, germ cells and sperm head abnormalities in the treated mice. Swiss mice were treated orally by gavage with 2,4-D at 1.7, 3.3 and 33 mg kg(-1)BW (1/200, 1/100 and 1/10 of LD(50)). 2,4-DCP was intraperitoneally (i.p.) injected at 36, 72 and 180 mg kg(-1)BW (1/10, 1/5, 1/2 of LD(50)). A significant increase in the percentage of chromosome aberrations in bone-marrow and spermatocyte cells was observed after oral administration of 2,4-D at 3.3 mg kg(-1)BW for three and five consecutive days. This percentage increased and reached 10.8+/-0.87 (P<0.01) in bone-marrow and 9.8+/-0.45 (P<0.01) in spermatocyte cells after oral administration of 2,4-D at 33 mg kg(-1)BW for 24 h. This percentage was, however, lower than that induced in bone-marrow and spermatocyte cells by mitomycin C (positive control). 2,4-D induced a dose-dependent increase in the percentage of sperm head abnormalities. The genotoxic effect of 2,4-DCP is weaker than that of 2,4-D, as indicated by the lower percentage of the induced chromosome aberrations (in bone-marrow and spermatocyte cells) and sperm head abnormalities. Only the highest tested concentration of 2,4-DCP (180 mg kg(-1)BW, 1/2 LD(50)) induced a significant percentage of chromosome aberrations and sperm head abnormalities after i.p. injection. The obtained results indicate that 2,4-D is genotoxic in mice in vivo under the conditions tested. Hence, more care should be given to the application of 2,4-D on edible crops since repeated uses may underlie a health hazard.     

Sequential anaerobic degradation of 2,4-dichlorophenol in freshwater sediments.

2,4-Dichlorophenol (2,4-DCP) was anaerobically degraded in freshwater lake sediments. From observed intermediates in incubated sediment samples and from enrichment cultures, the following sequence of transformations was postulated. 2,4-DCP is dechlorinated to 4-chlorophenol (4-CP), 4-CP is dechlorinated to phenol, phenol is carboxylated to benzoate, and benzoate is degraded via acetate to methane and CO2; at least five different organisms are involved sequentially. The rate-limiting step was the transformation of 4-CP to phenol. Sediment-free enrichment cultures were obtained which catalyzed only the dechlorination of 2,4-DCP, the carboxylation of phenol, and the degradation of benzoate, respectively. Whereas the dechlorination of 2,4-DCP was not inhibited by H2, the dechlorination of 4-CP, and the transformation of phenol and benzoate were. Low concentrations of 4-CP inhibited phenol and benzoate degradation. Transformation rates and maximum concentrations allowing degradation were determined in both freshly collected sediments and in adapted samples: at 31 degrees C, which was the optimal temperature for the dechlorination, the average adaptation time for 2,4-DCP, 4-CP, phenol, and benzoate transformations were 7, 37, 11 and 2 days, respectively. The maximal observed transformation rates for these compounds in acclimated sediments were 300, 78, 2, 130, and 2,080 micromol/liter(-1)/day(-1), respectively. The highest concentrations which still allowed the transformation of the compound in acclimated sediments were 3.1 m/M 2,4-DCP, 3.1 mM 4-CP, 13 mM phenol, and greater than 52 mM benzoate. The corresponding values were lower for sediments which had not been adapted for the transformation steps.     

2,4-二氯苯酚在土壤与河流底泥中降解动力学

以南京化学工业园内四柳河沿岸土壤与河流底泥为研究对象,通过土壤灭菌、温度与污染物初始浓度调控,研究了2,4-二氯苯酚在土壤与河流底泥中降解动力学及其影响因子。结果表明:微生物对2,4-二氯苯酚降解起主导作用,在45d内,非灭菌土壤和河流底泥的降解率分别是灭菌条件下的1.5～3倍、1.4～2.8倍,土壤和河流底泥中的2,4-二氯苯酚微生物降解量分别为0.128～0.599和0.113～0.718mg·kg-1,非灭菌处理半衰期时间短于灭菌处理;(10±1)℃～(30±1)℃范围内,随着温度的增高,2,4-二氯苯酚降解加快,在(30±1)℃土壤与河流底泥中残留量最小,分别为0.305和0.203mg·kg-1,半衰期也最短;土壤与河流底泥中的2,4-二氯苯酚均在其浓度为0.5mg·kg-1时降解最快,随着初始浓度的增加,2,4-二氯苯酚降解速度呈现递减趋势,半衰期增长。