43 resultados para anaerobic granular sludge

em Chinese Academy of Sciences Institutional Repositories Grid Portal


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本文从四川绵竹酒厂、成都市龙泉长安垃圾填埋场以及四川大学荷花池底的厌氧污泥中先后分离得到63株厌氧产氢菌,其中H-8、H-61、HC-10等16株产氢细菌产氢能力较高,HC-10的产氢能力最高,最大产氢量和最大产氢速率分别达到2840 ml H2/L培养基和25.39 mmol H2/g drycell·h,对HC-10进行生理生化鉴定和分子生物学鉴定,判定其为clostridium sp.,对HC-10的产氢条件进行了研究,结果表明,该菌的最适生长温度为35 ℃,最适生长初始pH为7,以葡萄糖为最佳碳源,以蛋白胨为最佳氮源,不利用无机氮源,其产氢发酵液相产物以乙醇和乙酸为主,其发酵类型属于乙醇型发酵。此外,以酒糟废液作为底物,进行了菌株HC-10的生物强化试验,研究表明,投加了HC-10的强化系统其产氢量比对照高出40.32%。 同时为了获得厌氧产氢菌的高效突变株,分别以产氢菌H-8和H-61为原始菌株进行微波诱变处理,对微波诱变参数进行了优化,考察了突变株的遗传稳定性、产氢特性及耐酸性。菌株H-8经过微波诱变得到5株高产氢突变株HW7、HW33、HW181、HW184、HW195,经多次传代表明HW195是稳定的高产突变株。突变株HW195具有较好的耐酸性,在pH值为2.8时仍能生长。通过间歇发酵实验,其最大产氢量和最大产氢速率分别达到2460 mL/L培养基和27.97 mmol H2/g drycell·h,比原始菌分别提高了50.75%和41.7%。菌株H-61经过微波诱变后选育得到的突变株HW-18,其最大产氢量和最大产氢速率分别达到2190 mL/L培养基和25.86 mmol H2/g drycell·h,比原始菌分别提高了23.03%和31.00%。 为了对比各种诱变方式对产氢菌产氢能力的影响,以厌氧产氢菌H-61为原始菌株,先后经亚硝基胍(NTG)、紫外(UV)诱变,选育得到1株高产突变株HCM-23。在葡萄糖浓度为10 g/L的条件下,其产氢量为3024 mL/L培养基,比原始菌株提高了69.89%;其最大产氢速率为33.19 mmol H2/g drycell·h,比原始菌株提高了68.14%。经过多次传代实验,稳定性良好。其发酵末端产物以乙醇和乙酸为主,属于典型乙醇型发酵。其最适产氢初始pH为6.5,最适生长温度为36 ℃,以蔗糖为最佳碳源。与原始菌株相比,突变株HCM-23的产氢特性发生了改变,如生长延滞期延长,可利用无机氮源等。 From anaerobic activated sludge, 16 strains of hydrogen producing bacteria were newly isolated. One of them named as HC-10 had the highest hydrogen producing capability, under the batch fermentative hydrogen production condition, the maximal hydrogen yield and hydrogen production rate was 2840 mL/L culture and 25.39 mmol H2/g drycell·h. It was identified as clostridium sp.HC-10 by 16S rDNA sequence analysis. Various parameters for hydrogen production, including substrates, initial pH and temperature, have been studied. The optimum condition for hydrogen producing of strain HC-10 were achieved as: initial pH 7.0, temperature 35 ℃, glucose as the favorite substrate, Moreover, using distiller's solubles wastewater as substrate, HC-10 strain was added in the biohydrogen producing system to research the bioaugmentation effection. The results showed that the hydrogen production of bioaugmentation system was 40.32% higher than the noaugmentation system. An anaerobic, hydrogen producing strain H-8 was irradiated by microwave to optimize the microwave mutagenesis condition, and to test the heredity, hydrogen-producing potential and aciduric of the mutants. An aciduric mutant named as HW195 with steady hydrogen-producing capability was obtained, which can grow at pH 2.8. Its capability of hydrogen production was tested in the batch culture experiments. The maximum hydrogen yield and hydrogen production rate was 2460 mL/L culture and 29.97 mmol H2/g drycell·h, which was 50.7% and 41.7% higher than those of the initial strain, respectively. When used the strain H-61 as original strain, a mutant named as HW18 was obtained. The maximum hydrogen yield and hydrogen production rate was 2190 mL/L culture and 25.86 mmol H2/g drycell·h, which was 23.03% and 31.00% higher than those of the initial strain, respectively. The results demonstrated that microwave mutagenesis could be used in the field of hydrogen producing microorganism. The hydrogen producing strain H-61 was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production capability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production capability. which was tested in the batch culture experiments. With the condition of 10 g/L glucose, its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L culture and 33.19 mmol H2/g drycell·h, 69.89%and 68.14% higher than that of the original strain, respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type, while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation were studied, including time, carbon source, nitrogen source, glucose concentration, glucose utilization, initial pH and incubation temperature had been studied, indicated the optimum condition of hydrogen production for the mutant HCM-23 as initial pH6.5, temperature 36 ℃, and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different, such as, the growth lag phase and the utilization of inorganic nitrogen source, etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

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本文从成都龙泉垃圾填埋场和宜宾造纸厂分离到耐酸性能优良的高温产甲烷菌RY3和中温产甲烷菌SH4,并将其与实验室现有的利用不同底物的产甲烷菌配伍组合成了复合菌剂。采用活性污泥作为固体附着物,研制出了固体产甲烷菌复合菌剂。 菌株RY3的pH耐受范围为5.5~10.5,最适生长pH 6.0~8.0。菌株RY3为革兰氏阳性,长杆状,多数单生,不运动;菌落浅黄色,形状近圆形;利用H2+CO2或甲酸盐作为唯一碳源生长,不利用乙酸盐,对氯霉素非常敏感。该菌最适生长温度为55℃~65℃,最适NaCl浓度为0~2%。根据形态和生理生化特性及16S rDNA序列分析将其初步定为热自养甲烷热杆菌(Methanothermobacter thermautotrophicus)。添加RY3菌液与仅添加厌氧污泥作为接种物相比一周内可使达到最大产甲烷速率所需时间缩短三分之二,甲烷总产量提高约1.8倍。菌株SH4的生长pH范围5.5~9.5,其对酸碱具有良好的适应性,培养3天后,在初始pH值为6.0~8.0的培养基中甲烷产量相差不大,且基本达到最大产量。SH4革兰氏染色阳性,短杆状,多数单生,不运动;菌落近圆形,微黄;利用H2+CO2或甲酸盐作为唯一碳源生长,不利用乙酸盐,对氯霉素非常敏感。SH4最适生长pH 为7.0,最适生长温度为35℃,最适NaCl浓度为0~1.5%。实验表明,添加SH4菌液与仅添加厌氧污泥作为接种物相比可使产甲烷启动时间缩短三分之一,甲烷总产量亦有大幅提高。从形态和生理生化特征以及16S rDNA序列分析表明SH4为嗜树木甲烷短杆菌(Methanobrevibacter arboriphilus)。 以活性污泥为附着物,与培养基和菌种经搅拌后厌氧发酵可得产甲烷菌固体复合菌剂。固体复合菌剂的pH耐受范围为5.5~9.5,温度耐受范围为15℃~65℃,表明其对环境的适应性较强。以猪粪为底物进行厌氧发酵,接种复合菌剂进行试验,以接种实验室长期富集的产甲烷厌氧污泥作为对照,在20℃时,发酵甲烷浓度与对照基本一致,但每日产气量优于对照,第15天时接种复合菌剂的发酵瓶每日产气量是对照的1.59倍;50℃时达到最大甲烷含量所需时间比对照缩短三分之二,三周内总产气量约为对照的2.7倍,甲烷总产量约为2.8倍。以不加接种物为对照,接种复合菌剂20℃时发酵甲烷含量达到50%约需2周,对照2周内甲烷含量最高仅为4.3%;50℃时接种复合菌剂发酵仅需约1周甲烷含量便可达50%,对照则至少需要2周。 In this paper, high-temperature Methanogen RY3 and middle-temperature SH4 were isolated from Chengdu Longquan refuse landfill and Yibin paper mill. They could be used to make compound inoculum that producing methane with the existing Methanogens utilized different substrate. With using anaerobic activated sludge be solid fixture, the process had been designed to produce solid compound inoculum. Strain RY3 possessed excellent capacity of acid and alkali-tolerant. The pH-tolerant scale of RY3 was 5.5~10.5 and its optimum pH value for growth was 6.0~8.0. RY3 was G+, long-rod shape, monothetic and nonmotile, the colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by RY3 as sole C-source, and it was very sensitive to chloramphenicol. Besides, strain RY3 grew fastest at 55℃~65 and 0℃~2% NaCl. Characteristics of modality and physiology with sequence analysis of the 16s rDNA gene of strain RY3 preliminarily showed that it was Methanothermobacter thermautotrophicus. The experiments indicated that the time which began to produce methane with the highest velocity could be shortened two third by adding RY3 in one week, and the total methane production also was 1.8 times than before. Strain SH4 possessed wide scale of growing pH(5.5~9.5)and excellent ability of acclimatizing itself to acid-alkali. The methane production had no apparent difference among those cultivated in different initial pH(6.0~8.0)after three days and equaled to the maximum production basically. Cells of SH4 were G+, short-rod sharp, monothetic and nonmotile. The colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by SH4 as sole C-source, and it was very sensitive to chloramphenicol. Besides, it grew fastest at pH 7.0,55 ℃~65 and 0℃~2% NaCl concentration. The experiment indicated the time that began to produce methane could be shortening one third by adding SH4. And the total methane production also rose apparently. Characteristic of modality and physiology with sequence analysis of the 16S rDNA gene of strain SH4 demonstrated it was Methanobrevibacter arboriphilus. The activated sludge was utilized as fixture, mixed with culture medium and inocolum, that the solid compound inoculum could be produced by anaerobic fermentation. The compound inoculum could grow between pH 5.5~9.5, 15℃~65. It demonstrated the compound inoculum ha℃ve great ability of adapting to circumstance. In the experiment that making pig manure be substrate and taking the anaerobic sludge producing methane that cultured in long term in laboratory to be comparison, the concentration of methane in fermentation added compound inoculum almost equal to the comparison at 20℃, but the volume of gas production could be a little higher. The gas production everyday inoculated compound inoculum was 1.59 times to comparison. The time that the concentration of methane to maximum could be shortening by two third by adding compound inoculum, and the total gas production was 2.7 times to comprison while the total methane production was 2.8 times. If take the no inoculum be the comprasion, anaerobic fermentation added compound inoculum made the concentration of methane to 50% in 2 weeks but the comparison only to 4.3% at 20℃. The time that the concentration of methane to 50% by adding compound inoculum only need 1 week, but the comparison need 2 weeks at 50℃.

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活性污泥法是目前世界上普遍应用的污水生物处理工艺,其在运行过程中产生大量的剩余污泥。由于剩余污泥处理费用巨大及污泥最终处置对环境具有潜在危害问题,污泥的处理和处置已经成为水处理领域关注的焦点。本文利用实验室筛选的溶胞菌群,在好氧消化的同时对污泥进行前处理,促进剩余污泥的破解与溶胞,再通过两相厌氧处理对污泥进行进一步消化,以研究投加溶胞菌对剩余污泥消化的影响。 本研究中溶胞菌污泥减量化技术分为两个部分,第一,污泥在溶胞菌作用下的好氧消化与污泥传统好氧消化的对比研究,利用取自成都三瓦窑污水处理厂剩余污泥,向好氧污泥消化反应器中投加溶胞菌,检测各项污泥指标,并通过同传统好氧污泥消化对比,以研究溶胞菌对污泥好氧消化的影响。第二, 经过溶胞菌处理后好氧消化的剩余污泥进行两相厌氧处理研究。通过建立好氧溶胞联合两相厌氧消化系统的来处理剩余污泥,并与相同条件运行的两相厌氧消化系统做对比,检测运行过程中系统中物质成分变化,研究了其处理能力和运行稳定性,探索了两相厌氧消化系统中的发酵类型差别,验证了好氧溶胞对剩余污泥的破解效果。 研究结果表明:污泥在溶胞菌作用下的好氧消化效果和消化效率均优于传统好氧消化。在溶胞菌群存在的情况下,剩余污泥的TSS和VSS去除率达到40%和53%,远高于传统好氧消化的12%和20%。污泥经过溶胞及好氧消化后,TCOD去除率达到54.4%。经过溶胞菌处理后的剩余污泥再进入两相厌氧处理系统,进入厌氧处理系统的剩余污泥的VSS/TSS比值约为0.62。在两相厌氧处理水力停留时间(HRT)为8d时,溶胞处理污泥厌氧消化后VSS去除率达到55.17%,对照组两相厌氧系统的VSS去除率平均值为18.53%。经过溶胞处理的两相厌氧系统的污泥减量了能力远高于对照组。两相厌氧系统的pH值和碱度说明系统运行较为稳定。产酸相的有机酸中乙酸含量高于丙酸和丁酸,说明发酵末端产物以乙酸为主。在20天的试验周期内,污泥溶胞处理后、两相厌氧系统产甲烷相产气量累积产气量为1.2L,对照组只有375ml。气体中甲烷含量都在55%左右。该研究结果表明,好氧溶胞对污泥有破解能力,溶胞处理对两相厌氧中产酸相水解污泥细胞有明显的促进作用,提高了产酸相的水解酸化能力和效率。该研究对于利用生物溶胞途径提高污泥消化效率具有重要意义。 The actived sludge process has been used more and more extensively, but the procedure will lead to a large quantity of excess sludge. The treatment of Excess activated sludge has becomes a focuses not only for it is a seriously negative effect on environment but also for the costly disposal comes subsequently. The cell lysing bacterium was keeped in our lab to joined in the digestion of the excess activated sludge which was carrying at the same time with pre-processing of sludge to investigated the influence of cell lysing bacterium on excess sludge. There are two part in the method of cell lysing bacterium digesting sludge technology, the first, comparison of excess sludge digestion between anaerobic Cell-lysing Pretreatment and Conventional Aerobic Process. The sludge which was collected from San Wanyao disposal plant in Chengdu was thrown into the aerobic process system with cell-lysing bacterium, then, the indexes were detected to compare the difference between the cell-lysing bacterium in aerobic process and the traditional method to determine the influence of cell-lysing bacterium on aerobic process ; The second, the research on the sludge which was pro-treated with cell-lysing and aerobic digestion in the diphase of anaerobic digestion system. The system of cell-lysing combined with diphase of anaerobic digesting was created to compare to the diphase of anaerobic digested system, the changes of mass constituent was detected to study the ability and steady of disposal. Moreover, the research explored the difference among the types of fermentation. The efficacious of aerobic process was been proved. The result shows that the digesting rate of aerobic process with cell-lysing bacterium was higher than the traditional process. The ratio of sludge is reach to 40%~53%, which was far more effectively than the traditional process rate of 12%~20%. The TCOD of sludge which was treated with cell lysing bacterium and Aerobic Process is reach to 54.4%. Then, the sludge was thrown into the diphase of anaerobic digesting system. VSS/TSS of sludge is 0.62, HRT is 6d, the reduction of VSS is reach to 40.8%. The pH and alkalinity indicate the steady running of the diphase anaerobic digest system. In the acerbity phasing, the content of acetic acid was more than butanoic acid and propanoic acid in organic acid, it is demonstrated that the main composition of final production of fermentation was Acetic Acid. During the 20d of experiment, methylhydride phasing of diphase anaerobic digest system produced 1.2L methylhydride, however, there is only 375ml in CK, the content of methylhydride in all gas phase was around the rate of 55%. The average ratio of VSS was 18.53% in CK diphase anaerobic digest system which was far more unavailable than the mass sludge rate of 55.17%. Results demonstrated that aerobic cell-lysing digested the sludge, the treat of cell-lysing could obviously promoted the hydrolyzeing of sludge cell in the acerbity phasing, which improved the ability and rate of hydrolization and acidification. This study is significant in inhenceing the rate of sludge digestion in the method of cell-lysing bacterium.

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The hydrogen production from the organic fraction of municipal solid waste (OFMSW) by anaerobic mixed culture fermentation was investigated using batch experiments at 37 degrees C. Seven varieties of typical individual components of OFMSW including rice, potato, lettuce, lean meat, oil, fat and banyan leaves were selected to estimate the hydrogen production potential. Experimental results showed that the boiling treated anaerobic sludge was effective mixed inoculum for fermentative hydrogen production from OFMSW. Mechanism of fermentative hydrogen production indicates that, among the OFMSW, carbohydrates is the most optimal substrate for fermentative hydrogen production compared with proteins, lipids and lignocelluloses. This conclusion was also substantiated by experimental results of this study. The hydrogen production potentials of rice, potato and lettuce were 134 mL/g-VS, 106 mL/g-VS, and 50 mL/g-VS respectively. The hydrogen percentages of the total gas produced from rice, potato and lettuce were 57-70%, 41-55% and 37-67%. 2008 International Association for Hydrogen Energy.

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Acid, alkali, heat-shock, KNO3 and control pretreatment methods applied to anaerobic sludge were evaluated for their ability to selectively enrich the marine hydrogen-producing mixed microflora. Seawater culture medium was used as the substrate. The hydrogen yield of pretreated microflora was higher than that of the un-pretreated control (P < 0.05). Among the pretreatment methods studied, heat-shock pretreatment yielded the greatest hydrogen production, which was 14.6 times that of the control. When the effect of initial pH on hydrogen production of heat-shock pretreated samples was studied, hydrogen was produced over the entire pH range (pH 4-10). The hydrogen yield peaked at initial pH 8 (79 mL/g sucrose) and then steadily decreased as the initial pH increased. Sucrose consumption was high at neutral initial pH. During the process of hydrogen production, pH decreased gradually, which indicated that the acquired microflora consisted of acidogenic bacteria.

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The effective refractive index of a kind of granular composite, which consists of granular metallic and magnetic inclusions with different radius embedded in a host medium, is theoretically investigated. Results show that for certain volume fractions of these two inclusions, the negative permittivity peak shifts to low frequency and the peak value increases with increasing radius ratio of the radius of magnetic granulae to that of metallic granulae. Simultaneously, peak value of permeability decreases with the radius ratio, and value peak shifts to high frequency with increasing volume fraction of magnetic inclusion. Therefore, the radius ratio can affect the effective refractive index considerably, and it is found that by adjusting the radius ratio, the refractive index may change between negative and positive values for certain volume fractions of the two inclusions. (c) 2005 Elsevier B.V. All rights reserved.

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Oxidation-reduction properties of surface sediments are tightly associated with the geochemistry of substances, and reducing organic substances (ROS) from hydrophytes residues may play an important role in these processes. In this study, composition, dynamics, and properties of ROS from anaerobic decomposition of Eichhornia crassipes (Mart.) Solms, Potamogenton crispus Linn, Vallisneria natans (Lour.) Hara, Lemna trisulca Linn and Microcystis flos-aquae (Wittr) Kirch were investigated using differential pulse voltammetry (DPV). The type of hydrophytes determined both the reducibility and composition of ROS. At the peak time of ROS production, the anaerobic decomposition of M. flos-aquae produced 6 types of ROS, among which 3 belonged to strongly reducing organic substance (SROS), whereas there were only 3-4 types of ROS from the other hydrophytes, 2 of them exhibiting strong reducibility. The order of potential of hydrophytes to produce ROS was estimated to be: M. flos-aquae > E. crassipes > L. trisulca > P. crispus approximate to V. natans, based on the summation of SROS and weakly reducing organic substances (WROS). The dynamic pattern of SROS production was greatly different from WROS. The total SROS appeared periodic fluctuation with reducibility gradually weakening with incubation time, whereas the total WROS increased with incubation time. Reducibility of ROS from hydrophytes was readily affected by acid, base and ligands, suggesting that their properties were related to these aspects. In addition to the reducibility, we believe that more attention should be paid to the other behaviors of ROS in surface sediments.

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Strongly reducing organic substances (SROS) and iron oxides exist widely in soils and sediments and have been implicated in many soil and sediment processes. In the present work, the sorptive interaction between goethite and SROS derived from anaerobic decomposition of green manures was investigated by differential pulse voltammetry (DPV). Both green manures, Astragaltus sinicus (Astragalus) and Vicia varia (Vicia) were chosen to be anaerobically decomposed by the mixed microorganisms isolated from paddy soils for 30 d to prepare different SROS. Goethite used in experiments was synthesized in laboratory. The anaerobic incubation solutions from green manures at different incubation time were arranged to react with goethite, in which SROS concentration and Fe(II) species were analyzed. The anaerobic decomposition of Astragalus generally produced SROS more in amount but weaker in reducibility than that of Vicia in the same incubation time. The available SROS from Astragalus that could interact with goethite was 0.69 +/- 0.04, 0.84 +/- 0.04 and 1.09 +/- 0.03 cmol kg(-1) as incubated for 10, 15 and 30 d, respectively, for Vicia, it was 0.12 +/- 0.03, 0.46 +/- 0.02 and 0.70 +/- 0.02 cmol kg(-1). One of the fates of SROS as they interacted with goethite was oxidation. The amounts of oxidizable SROS from Astragalus decreased over increasing incubation time from 0.51 +/- 0.05 cmol kg(-1) at day 10 to 0.39 +/- 0.04 cmol kg(-1) at day 30, but for Vicia, it increased with the highest reaching to 0.58 +/- 0.04 cmol kg(-1) at day 30. Another fate of these substances was sorption by goethite. The SROS from Astragalus were sorbed more readily than those from Vicia, and closely depended upon the incubation time, whereas for those from Vicia, the corresponding values were remarkably less and apparently unchangeable with incubation time. The extent of goethite dissolution induced by the anaerobic solution from Vicia was greater than that from Astragalus, showing its higher reactivity. (c) 2008 Published by Elsevier Ltd.

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The bacterial diversity of activated sludge from submerged membrane bioreactor (SMBR) was investigated. A 16S rDNA clone library was generated, and 150 clones were screened using restriction fragment length polymorphism (RFLP). Of the screened clones, almost full-length 16S rDNA sequences of 64 clones were sequenced. Phylogenetic tree was constructed with a database containing clone sequences from this study and bacterial rDNA sequences from NCB1 for identification purposes. The 90.6% of the clones were affiliated with the two phyla Bacteroidetes (50%) and Proteobacteria (40%), and beta-, -gamma-, and delta-Proteobacteria accounted for 7.8%, 28.1%, and 4.7%, respectively. Minor portions were affiliated with the Actinobacteria and Firmicutes (both 3.1%). Only 6 out of 64 16S rDNA sequences exhibited similarities of more than 97% to classified bacterial species, which indicated that a substantial fraction of the clone sequences were derived from unknown taxa. Rarefaction analysis of operational taxonomic units (orrUs) clusters demonstrated that 150 clones screened were still insufficient to describe the whole bacterial diversity. Measurement of water quality parameter demonstrated that performance of the SMBR maintained high level, and the SMBR system remained stable during this study.

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The graphite electrode sludge was sampled from a huge chloralkali plant in central China. The total level of PCDD/F was found as high as 378.85 mu g/kg sludge (dry weight). The patterns of PCDD/F in each homologue indicated the predominance of tetra- to octa-chlorinated PCDFs, Furthermore, the toxic 2,3,7,8-substituted PCDFs constituted over 80% of the total PCDFs in the sludge and the corresponding PCDDs were only at 15 mu g/kg level. The calculated value of the international toxic equivalence (I-TEQ) in sludge was 21.65 mu g/kg sludge (dry weight). This typical "dioxin chloralkali pattern" was apparently identified in the sediments near the effluent outlet of the chloralkali plant.