Evaluation of the OECD activated sludge,respiration inhibition test

The OECD Activated Sludge, Respiration Inhibition Test was evaluated as a method for assessing the potential impact of chemicals on wastewater treatment systems. Reproducibility of the test method was examined with respect to variation in oxygen consumption rates measured for a series of reactions set under identical conditions, and the variability in IC50 values estimated for three reference compounds. Several statistical procedures were also evaluated for analyzing activated sludge inhibition data. The test method was subsequently used to examine the effects of a wide variety of inorganic and organic chemicals on activated sludge respiration rates.     

Evaluation of the toxicity of substances to be assessed for biodegradability

The ability to distinguish between true non-biodegradability, and inhibition of biodegradability due to the toxic effects of the compound tested present problems. It results in possible false identification of substances as non-biodegradable.Six chemicals were evaluated in a variety of tests to assess their toxicity to aerobic sludge organisms. The tests employed were: BOD₅ and Closed Bottle inhibition tests, inhibition of respiration of activated sludge, growth inhibition of activated sludge, light emission from (MICROTOX test) and the repetitive die-away test (RDA). Results from these tests were compared with results obtained from a number of ready biodegradability tests using the compounds listed above at anticipated non-toxic and toxic concentrations.No test method evaluated consistently forecast toxicity due to the chemicals tested. The MICROTOX and nitrification inhibition tests were too sensitive. A combination of sludge respiration rate and/or growth tests seem most appropriate. To avoid toxicity to the sludge, biodegradability testing should be made at 10% of the EC₅₀ value. Compounds with an EC₅₀ value greater than 300 mg/l are unlikely to be toxic in ready biodegradability tests. Compounds with an EC₅₀ value of less than 20 mg/l may pose problems necessitating the use of the stringent closed bottle test, or the use of C¹⁴ labelled test materials. Compounds with an intermediate EC₅₀ that is between 20–300 mg/l, need to be evaluated at a range of concentrations in biodegradability tests, or may need to be evaluated carefully to define the precise no-effect level. Procedure recommendations to distinguish between the inhibition of biodegradability and inertness of the test substance are made.     

Effects of nano-copper(II) oxide and nanomagnesium oxide particles on activated sludge

Effects of nano-copper(II) oxide (nano-CuO) and nanomagnesium oxide (nano-MgO) particles on activated sludge endogenous respiration (aerobic digestion), biochemical oxygen demand (BOD) biodegradation, and nitrification were investigated through respiration rate measurement. For comparison, the effects of Cu(II) and Mg(II) ions on activated sludge were also studied. Results indicated that soluble Cu(II) has half maximum inhibitory concentration (IC50) values of 19, 5.5, 53, and 117 mg Cu/L for endogenous respiration, BOD biodegradation, ammonium oxidation, and nitrite oxidation, respectively. However, nano-CuO only inhibited BOD biodegradation at 240 mg Cu/L or more, and its associated toxicity was primarily caused by soluble Cu(II). In contrast, soluble Mg(II) was not toxic to activated sludge in the experimental concentration range, but nano-MgO inhibited BOD biodegradation and nitrification with IC50 values of 70 and 143 mg Mg/L, respectively. Further study indicated that the toxicity of nano-MgO resulted primarily from increased pH following MgO hydrolysis.     

Effect of arsenic and mercury speciation on inhibition of respiration rate in activated sludge systems

The inhibition effect of arsenite, As(III), arsenate, As(V), inorganic mercury, Hg(II) and methylmercury, MeHg, on the respiration rate of activated heterotrophic sludge microorganisms was evaluated. As(III) and MeHg were much more toxic to activated sludge than As(V) and Hg(II) respectively. The effect of various experimental parameters on the toxicity, such as sludge age, concentration of suspended solids and exposure time, was investigated. An increase of sludge age or the concentration of suspended solids reduces the observed inhibition. Longer exposure seems to dramatically increase the inhibition of As(III), MeHg and Hg(II) during the first hours of exposure, while the later inhibition increases at a slower rate. On the contrary, in the presence of As(V), 24 hours after exposure, the respiration rate was similar to that of the control biomass.     

Toxicity of metals and organic chemicals evaluated with bioluminescence assays

The development of a bioluminescent sensor organism (Shk1) that was created for assessing wastewater toxicity was reported several years ago. In order to establish a test battery to better characterize wastewater toxicity, additional luminescent sensor organisms were later created. The present study focused on one promising candidate (PM6), a Pseudomonas spp. strain, because of its high level of luminescence compared to that of other newly created organisms. Using a batch toxicity testing protocol, the toxicity of 7 metals and 25 organic compounds was evaluated with the PM6 and Shk1 assays. Results indicated that the toxicity data of the PM6 and the Shk1 assays were correlated, and no assay appeared to be particularly more sensitive to a group of toxicants than the other assay. The results of the PM6 and Shk1 assays were further evaluated by comparing with the results of the Vibrio fischeri luminescence inhibition assay and activated sludge inhibition assays. Data suggested that PM6 and Shk1 more closely represented activated sludge organisms than V. fischeri. The suitability of using PM6 and Shk1 for assessing wastewater toxicity on activated sludge, both individually and in a test battery, was discussed.     

废水可生物降解COD组分2种表征方法的比较

利用自行开发的混合呼吸速率测量仪在接种污泥的条件下对重庆某城市污水处理厂污水进行了呼吸速率测试（短期BOD测试，以呼吸速率测量重新进入内源呼吸阶段为结束）；同时应用美国产BI-2000电解质呼吸仪在不接种污泥的条件下对该污水进行了BOD测试（长期BOD测试，理论上以污水中所有有机物矿化为结束）。对2种测试方法及其结果进行了比较，结果表明，2种方法得到的BCOD存在很大差异，短期BOD测试方法得到的结果仅为长期BOD测试方法得到的结果的40％～60％。通过批式呼吸测量方法测定了原废水中的活性异养微生物浓度XH(0)，结果表明，XH(0)与BCOD_st之和与BCOD_lt比较接近，两者之比在0.88～1.02之间，平均值为0.94。     

抗生素类污染物对活性污泥生物毒性测定方法的筛选与评价

尽管衡量化学物质生物毒性的标准方法很多,但关于环境中抗生素类污染物的生物毒性及其对污泥活性影响的科学数据较少。以磺胺和四环素两类作用机理和作用谱带不同的抗生素为研究对象,分别用发光细菌法、脱氢酶活性法、生长抑制法及呼吸速率法进行了抗生素类污染物生物毒性测定方法的筛选与评价。结果表明,发光细菌标准方法中30min的作用时间太短,延长作用时间不仅导致各种抗生素的EC50值大幅度降低,同时毒性排列顺序也发生了改变;以活性污泥为对象的脱氢酶活性和呼吸速率抑制实验的EC50值与抗生素类物质对敏感致病菌的MICs相比异常高,不适宜于作为单独方法准确评估抗生素类污染物的生物毒性。生长抑制实验中,活性污泥混合菌种增殖生长对磺胺类抗生素敏感,而假单胞杆菌对四环素类抗生素敏感。不同方法测定抗生素毒性的灵敏度顺序是发光细菌(24 h)>生长抑制(7 h)>呼吸速率(24 h)>脱氢酶活性(24 h)。用标准方法评价抗生素类污染物的生物毒性,可能导致对抗生素排放到水环境中所带来的风险估计不足。     

Activated sludge inhibition by chemical stressors--a comprehensive study.

The effects of shock loads of 1-chloro-2,4-dinitrobenzene (CDNB); cadmium; 1-octanol; 2,4-dinitrophenol (DNP); weakly complexed cyanide; pH 5, 9, and 11; and high ammonia levels on activated sludge biomass growth, respiration rate, flocculation, chemical oxygen demand removal, dewaterability, and settleability were studied. For all chemical shocks, except ammonia and pH, concentrations that caused 15, 25, and 50% respiration inhibition were used to provide a single pulse shock to sequencing batch reactor systems containing a nitrifying or non-nitrifying biomass. Cadmium and pH 11 shocks were most detrimental to all processes, followed by CDNB. The DNP and cyanide primarily affected respiration, while pH 5, pH 9, octanol, and ammonia did not affect the treatment process to a significant extent. A chemical source-process effect matrix is provided, which we believe will aid in the development of methods that prevent and/or attenuate the effects of toxic shock loads on activated sludge systems.     

A comparison of five rapid direct toxicity assessment methods to determine toxicity of pollutants to activated sludge

Five rapid direct toxicity assessment methods were used in three European partner countries to determine the toxicity of single toxicants, mixed toxicants and real industrial wastes. The final aim was to protect microbial degradation of organic wastes in biological treatment processes and hence enhance the quality of treated effluents to be discharged to the environment. Nitrification inhibition, Respirometry, Adenosine triphosphate luminescence and Enzyme inhibition were tested utilising activated sludge as the testing matrix. The Vibrio fischeri toxicity test was used as a surrogate to compare the various microbial bioassays. The IC50 (toxicant concentration eliciting a 50% inhibitory effect) was determined for a number of pollutants including single toxicants Cd, Cr, Cu, Zn, 3,5-dichlorophenol, toluene and linear alkylbenzenesulphonate (LAS); a standard mixture of metals and LAS; a standard mixture of organics and LAS, and 16 industrial effluents. The V. fischeri bioassay was also chosen in order to assess quality control of toxicant preparation during testing in the different laboratories of the partner countries. Comparisons of sensitivity, cost of implementation, cost per test, relevance, and ease of use were made. The most sensitive bioassays were V. fischeri and Nitrification inhibition, however, this depended in the main on the pollutant and mixtures tested. It is recommended that during assessment of wastewater toxicity a suite of tests be used rather than reliance on one particular test.     

Evaluation of a respirometric test method to determine the heterotrophic yield coefficient of activated sludge bacteria

The heterotrophic yield coefficient of activated sludge bacteria is an important parameter in the field of wastewater degradation kinetics and the determination of degradation kinetics of defined chemicals. With the help of easily biodegradable organic compounds like glucose and acetate the respiration kinetics of degrading activated sludge bacteria was studied. The results of the respiration analysis were transformed to examine the degradation and respiration kinetics of municipal wastewaters at different food to microorganism ratios (F/M ratios). It was proved that the heterotrophic yield coefficient of aerobically degrading activated sludge bacteria could reliably be determined and that the heterotrophic yield coefficient was independent of the F/M ratio over a wide range.     

Evaluation and further development of the activated sludge respiration inhibition test

The activated sludge respiration inhibition test is an important bacterial test system for the determination of bacterial toxicity of chemical compounds. The exposure period recommended by OECD 209 and ISO 8192 protocols is 30 and 180 min. A modified version of the test was developed which allowed a prolonged incubation period of 27 h to enhance the possibilities of the test system. The test system with the prolonged incubation time was evaluated by the recommended reference compound 3.5-dichlorophenol and showed an EC50 of 6.3 mgl(-1) with a coefficient of variation of 12.7%. Furthermore, the use of an open test system was evaluated showing a comparable toxicity but a higher coefficient of variation than the closed test system. A storage of activated sludge for several days accompanied by a daily feed with OECD nutrient solution should be avoided, for it can cause a decreased sensitivity of the inoculum. Different statistical fit procedures were tested indicating that Weibull fit procedures were superior for extended data sets covering a wide range of concentrations whereas Gamma and Probit fits were appropriate for smaller data sets mainly restricted to the linear part of the dose response curve.     

Determination of detoxification to Daphnia magna of four pharmaceuticals and seven surfactants by activated sludge

Pharmaceuticals are bioactive compounds generally resistant to biodegradation, which can make them problematic when they are released into nature. The use pattern for pharmaceuticals means that they are discharged into water via sewage treatment plants. Also surfactants are discharged through sewage treatment plants, primarily due to their use in detergents and shampoos and other cleaners. In this study the acute toxicity to Daphnia magna of four pharmaceuticals (ciprofloxacin, ibuprofen, paracetamol and zinc pyrithione) and seven surfactants (C8 alkyl glucoside, C6 alkyl glucoside, sodium caprylimidiopropionate, tallow-trimethyl-ammonium chloride, potassium decylphosphate, propylheptanol ethoxylate and alkylmonoethanolamide ethoxylate) was determined. Abiotic (without activated sludge bacteria) and biotic (with activated sludge bacteria) detoxification was also determined. The 24-h EC50s ranged from 2 μg L(-1) for the most toxic substance (zinc pyrithione) to 2 g L(-1) for the least toxic compound (C6 alkyl glucoside). Detoxification rates determined as the ratio between initial EC50 and EC50 after 1 week in water with activated sludge bacteria ranged from 0.4 (paracetamol) to 13 (zinc pyrithione). For most of these chemicals detoxification rate decreased after 1 week, but for one (alkylmonoethanolamide ethoxylate) it increased from about 2 to 30 times after 2 weeks. Many of these chemicals were "detoxified" also abiotically at about the same rate as biotically. Further studies are needed to determine the degradation products that were precipitated (aggregated) for some of the tested chemicals. Altogether, this study has shown that there are large differences in toxicity among chemicals entering sewage treatment plants, but also that the detoxification of them can differ. Therefore, the detoxification should receive more attention in the hazard and risk assessment of chemicals entering sewage treatment plants.     

Enhancement of batch waste activated sludge digestion by microwave pretreatment.

Batch anaerobic digesters were used to stabilize microwave (MW)-irradiated waste activated sludge (WAS). A low temperature range (50-96 degrees C) MW irradiation was applied. Effects of pretreatment temperature (T) and intensity (I), concentration (C) and percentage of sludge pretreated (PT) were investigated in a multilevel factorial statistical design containing 54 mesophilic batch reactors by monitoring cumulative biogas production (CBP). Variance analysis (ANOVA) determined that the most important factors affecting WAS solubilization were temperature, intensity, and sludge concentration. Improvements in CBP from WAS were significantly affected by sludge percentage pretreated, temperature, and concentration. Pretreatment resulted in 3.6 +/- 0.6 and 3.2 +/- 0.1 fold increases in soluble to total chemical oxygen demand (SCOD/TCOD) at high and low sludge concentrations, respectively. WAS, microwaved to 96 degrees C, produced the greatest improvement in CBP with 15 +/- 0.5 and 20 +/- 0.3% increases over controls after 19 d of digestion at low and high WAS concentrations. Dewaterability of microwaved sludge was enhanced after anaerobic digestion.     

ECOSUNIDE工艺活性污泥中自养菌浓度的测定与分析

设计并利用在差压仪中密闭投加基质的实验方法,根据ASM模型中自养菌和硝化过程耗氧速率的关系,对比研究了ECOSUNIDE工艺和其他4个污水处理厂曝气池混合液中自养菌的浓度。研究表明,在不考虑自养菌增长和衰减情况下,ECOSUNIDE工艺混合液自养菌生物浓度为48.15 mg COD/L,该结果显著高于其他受试污水处理厂的混合液自养菌生物浓度(17.2～37.5 mg COD/L),表明ECOSUNIDE工艺在提高活性污泥中自养菌生物浓度上有较强的优势。设计的实验方法可以简便、有效地测定用于ASM模型中的自养菌浓度。     

Biodegradation in laboratory activated sludge plants and aquatic toxicity of herbicides

The biodegradation and the aquatic toxicity of four herbicides (isoproturon, terbuthylazine, mecoprop, metamitron) were investigated. Laboratory activated sludge plants were used for biodegradation experiments. The biodegradation of mecoprop reached nearly 100%, the other herbicides were not eliminated by biodegradation. The acute Daphnia magna 24-h assay, the algal 72-h inhibition test, and the recently developed lemna growth inhibition 7-d test were applied to evaluate the biological effects of herbicides as original substances. EC 50 and EC 10 values were determined. Algal and lemna test show that isoproturon and terbuthylazine are both much more toxic than mecoprop and metamitron. Daphnids are generally less sensitive against herbicides than plants. Biodegradation and toxicity test were coupled for mecoprop to assess biological long-term effects of possible biodegradation products of this herbicide. The effluents of the laboratory activated sludge units were used in toxicity tests (Daphnia magna 21-d reproduction test, lemna growth inhibition 7-d test). No inhibiting effect on the tested organisms was observed.     

Hydraulic residence time effects on performance of an activated sludge unit treating wastewater containing dichlorophenol.

Wastewaters containing chlorophenol compounds are difficult to treat by biological means because of the toxic effects of those compounds on microorganisms. To investigate the adverse effects of chlorophenols on microorganisms, synthetic wastewater containing 2,4 dichlorophenol (DCP) was biologically treated in an activated sludge unit at different hydraulic residence times (HRTs) between 5 and 40 hours, whereas the feed chemical oxygen demand (COD), DCP concentrations, and sludge age were kept constant at 2500 +/- 50 mg/L, 150 mg/L, and 20 days, respectively. The resazurin method based on dehydrogenase activity was used for assessment of the feed and effluent wastewater toxicity. Percent COD, DCP, and toxicity removals increased, and the effluent COD, DCP, and toxicity levels decreased with increasing HRT. Biomass concentration in the aeration tank increased with increasing HRT because of low levels of DCP at high HRT levels, resulting in high COD, DCP, and toxicity removals. The sludge volume index decreased with increasing HRT, yielding well-settling organisms as a result of low levels of toxicity and high concentrations of active cells. Percent DCP and COD removals decreased with increasing specific DCP loading rate. The rates of DCP and COD removals showed a maximum at a low DCP concentration of 6 mg/L in the aeration tank, corresponding to a 25-hour HRT.     

Diffusion of oxygen through activated sludge flocs: experimental measurement, modeling, and implications for simultaneous nitrification and denitrification.

Diffusion of dissolved oxygen through activated sludge flocs was studied, as it represents a potential mechanism for simultaneous nitrification and denitrification in activated sludge systems. Dissolved oxygen profiles through six floc particles collected at different times from a full-scale activated sludge plant demonstrated that that the dissolved oxygen concentration declines through all floc particles. For larger floc particles (2-mm diameter and greater), the dissolved oxygen concentration reached near-zero values at depths depending on process operating conditions. A mathematical model based on diffusion of dissolved oxygen, organic substrate (methanol), ammonia, nitrite, and nitrate through a spherical floc and consumption of dissolved oxygen by heterotrophs and autotrophs accurately predicted the dissolved oxygen profile and required adjustment of only one model parameter--the concentration of heterotrophs. A different dissolved oxygen decline pattern was exhibited for the smaller floc particles characterized, with the dissolved oxygen reaching a non-zero plateau toward the center of the floc. This pattern was not reproduced with the mathematical model developed and suggests that additional mechanisms are responsible for the transport of dissolved oxygen into the center of these flocs. Implications of these results regarding the occurrence of simultaneous nitrification and denitrification include consideration of the factors that affect floc size and distribution (simultaneous nitrification and denitrification is maximized with larger floc particles), coupling of the International Water Association (London) activated models to predict activated sludge composition with diffusion models to consider intrafloc effects, and the effects of substrate diffusion on the apparent half-saturation constant for various substrates in activated sludge systems.     

投加沸石粉活性污泥搅拌参数优化及其对离子交换速率的影响研究

在0.3 m×0.3 m×0.36 m方形搅拌槽中初步探讨了不同的搅拌器类型、安装高度以及曝气强度对投加了沸石粉的高浓度活性污泥悬浮状态的影响,并根据固液悬浮理论确定了最佳的搅拌参数.同时,进行了不同搅拌速率下沸石对污水中氨氮的离子交换动力学研究.结果表明在无曝气的情况下,选用三叶推进式搅拌器,搅拌器离底高度为1/3搅拌器直径时所需要的临界搅拌速率最低.曝气有助于反应池中污泥的悬浮,对于30 g/L的污泥,当曝气强度由10 m3/(m2·h)增大1倍时,临界搅拌速率降低了50%.搅拌速率为120 r/min时,沸石对污水中氨氮的离子交换速率受膜扩散过程控制,当其增大为180 r/min时,其控制步骤为粒内扩散,沸石粉活性污泥达到完全均匀悬浮的临界搅拌速率不足以满足传质的要求.需增大一定程度.     

Applications of the deep-shaft activated sludge process in wastewater treatment

The deep-shaft activated sludge process is a unique modification of the activated sludge system. The main objective of this system is to increase the amount of dissolved oxygen available for biological activity. This can be achieved by increasing the rate of oxygen transfer from the gas phase to the liquid phase. The deep-shaft configuration increases the partial pressure of oxygen, thereby causing a high saturation concentration in the reactor. In the deep-shaft process, owing to high oxygen availability, a higher organic loading can be accommodated with a comparatively low air supply. This reduces the energy and area requirements and lowers the overall cost of treatment. This technology has been successfully applied for the high-rate treatment of strongly polluted wastewater, as well as for the treatment of wastewater containing toxic or slowly biodegradable pollutants. This paper presents different applications of the deep-shaft activated sludge process, along with their relative performances.     

重金属对活性污泥微生物毒性的比较研究

采用发光细菌毒性、活性污泥脱氢酶毒性、硝化抑制毒性3种方法测定Hg、Cd、Zn、Pb 4种重金属对活性污泥微生物的毒性,并对测定结果进行了比较.结果表明,发光细菌毒性测定方法的灵敏度最高,测得的重金属半数有效浓度(ECSO)最低,4种重金属对发光细菌发光强度的抑制程度由大到小顺序依次为Hg>Cd>Zn>Pb;活性污泥脱氢酶毒性和硝化抑制毒性的测定结果与发光细菌毒性测定结果相比,灵敏度相对较低,测得的重金属ECSO相对较高,测得的活性污泥脱氢酶活性的抑制程度由大到小顺序依次为Cd>Hg>Zn>Pb,与测得的活性污泥硝化速率抑制程度大小顺序一致.但2者测得的EC50有所差别.为了更准确的判定重金属对活性污泥微生物的毒性影响.至少应取不同的重金属毒性终点指示指标做一组毒性实验,而不能以发光细菌毒性测定结果作为唯一的判定依据,这可能会过分夸大重金属对污水处理工艺的冲击能力,导致污水处理成本无谓增加.