Acclimation of activated sludge to degrade toxic levels of 2,4-dinitrophenol.

Biodegradation of 75 and 100 mg/l of 2,4-dinitrophenol (DNP) by activated sludge acclimated in a Sequencing Batch Reactor (SBR) consistently required less than 6 hours although a lag at the beginning of every 48-hour SBR cycle was observed. Other investigators have reported that DNP levels of 100 mg/l and higher are significantly toxic even to acclimated bacteria. The activated sludge acclimated to 75 mg/l initial DNP had over 100 times the DNP-degrading bacteria than an SBR acclimated to 10 mg/l DNP, although the MLSS concentration in both reactors was similar. Results suggest that two mechanisms are responsible for activated sludge acclimation to toxic levels of DNP: maintenance of DNP-degrading biomass sufficiently large to reduce initial DNP to non-toxic levels, allowing for subsequent rapid degradation; and extension of the aeration period well beyond the time required for degradation to prevent gradual accumulation of any by-product which might also be toxic.     

The use of chemical uncouplers for reducing biomass production during biodegradation

Reducing the energy available for anabolism of cell mass was identified as a method to minimise disposal requirements of excess biomass produced in the activated sludge process. A model system consisting of Pseudomonas putida, maintained in a chemostat, was employed to investigate biomass production in the presence of the energy dissipating protonphore, p-nitrophenol (pNP). The efficiency of biomass production was reduced by up to 62% when the feed was supplemented with 100 mg pNP.1⁻¹ with a simultaneous increase in the specific substrate uptake rate. The data obtained have been analysed to reveal maintenance energy requirements and true growth yields. Cells were found to satisfy their maintenance energy requirements prior to utilising energy in anabolism. Decreases in pH alone had no effect on biomass production, but caused additional protonphore induced reduction of biomass production. A pH 6.2 the efficiency of biomass production was reduced by up to 77% when the feed was supplemented with 100 mg pNP.1⁻¹.     

Substrate utilization characteristics of the predominant filamentous and floc-forming bacteria isolated from a chemical fiber factory wastewater treatment plant

To supply theoretical verification of the function of a selector to control aerobic activated sludge bulking in the wastewater treatment plant for a chemical fiber factory in Taiwan, the filamentous and floc-forming bacteria in the aeration tank in the full-scale plant were examined microscopically and isolated. The kinetic characteristics of filamentous and floc-forming bacteria were also investigated.The predominant filamentous organism was Sphaerotilus natans. In addition to this organism, 21 strains of non-filamentous bacteria were isolated using the plate count method. These included Aeromonas jandaei DNA group 9, Acinetobacter johnsonii/genospecies 7, Bacillus pasteurii and Bacillus sp. (using the Biolog identification system). Nine strains showed the ability to form flocs when cultivated in glucose mineral salts medium.With glucose and acetate as sole substrates, the 4 floc-forming bacteria tested showed different substrate utilization characteristics. The 4 strains could be divided into 3 groups. The first group was the substrate degrading bacteria, the second group was the acid degrading bacteria and the other strains were those that had the highest substrate degradation rates at low substrate concentrations (below 800 mg/l). None of the floc-formers could utilize ethylene glycol, which is the major wastewater component. The kinetic characteristics of filamentous bacterium S. natans (Km = 4.0 mg glucose/l, Vm = 0.43 μl O₂/l) and the flocforming bacterium Aeromonas jandaei DNA group 9 (Km = 34.8 mg glucose/l, Vm = 0.59 μl O₂/l) provided information for selector design.     

Influence of temperature and sludge loading on activated sludge settling,especially on Microthrix parvicella

During recent years modern full scale wastewater treatment plants with biological nitrification, denitrification and phosphorus removal have had increasing problems with foam formation on the surfaces of aerobic tanks and with bulking activated sludge. The results of a survey in 1995 (Kunst and Knoop, 1996) showed that most often the filamentous bacterium Microthrix parvicella is responsible for these problems. Up to today there is only little knowledge about its selection criteria in activated sludge. Therefore several expenments were done in full scale activated sludge plants and in laboratory systems under defined conditions to investigate the influence of low (< 0.1 kg/(kg·d)) and high (≤ 0.2 kg/(kg·d)) BOD₅-sludge loading rates on the growth and morphology of M. parvicella and the settlement of activated sludge. Furthermore the influence of temperatures of 5°C, 12°C and 20°C on the growth of M. parvicella was investigated. It was shown that M. parvicella grows at low BOD₅-sludge loading rate and low temperature and is the main causative organism of bulking and foaming sludge in nutrient removal plants. On the basis of this investigation it was concluded that the growth of M. parvicella and the settling problems of the activated sludge resulting from excessive growth of this filament will always appear in modern municipal wastewater treatment plants with BOD₅-sludge loading rate ≤ 0.1 kg/(kg·d) especially under low temperature conditions.     

Detection of filamentous and nitrifying bacteria in activated sludge with 16S rRNA probes

Activated sludge samples from wastewater treatment plants from potato starch and starch derivatives factory and from a municipal sewage treatment plant were analyzed with DNA probes specific for several filamentous bacteria. It was found that Haliscomenobacter hydrossis, Sphaerotilus natans, Thiothrix sp. and Eikelboom Type 021N were common in the activated sludges. Fluorescent in situ Hybridization (FISH) analysis could detect more types of sheathed bacteria and yielded a more accurate quantification of bacteria than conventional microscopy.In a pilot and a full scale wastewater treatment plant (WWTP) clear correlations were found between the SVI and growth of a Sphaerotilus natans and a Thiothrix sp. Addition of chlorine to the bulking sludge resulted in an improved SVI of the sludge but only damaged filamentous cells outside the floc. Nitrification was measured with substrate depletion and FISH analysis. Signal interpretation of FISH analysis was demonstrated both manually and with automated image analysis.     

Multi-component kinetics of activated sludge treatment of bleached kraft mill effluent.

This study investigated the discrepancies between the BOD removal rates measured during short term assays and those measured during continuous activated sludge treatment of bleached kraft mill effluent (BKME). A combination of batch tests and fed batch tests with oxygen uptake rate (OUR), chemical oxygen demand (COD), biochemical oxygen demand (BOD), and mixed liquor volatile suspended solids (MLVSS) measurements were used to characterize the degradation rates for the activated sludge treatment of BKME and to divide the soluble readily biodegradable substrate into two to five separate fractions based on biodegradation rates. The removal rates varied by over an order of magnitude between the most readily degradable substrates (1 x 10(-3) mg COD/mg MLVSS minute), and the more slowly degradable substrates (2 x 10(-5) mg COD/mg MLVSS minute). If the readily biodegradable fraction of BKME was modeled as one substrate, initial rate kinetic measurements from batch tests were heavily influenced by the fractions with the greatest degradation rates, while any remaining BOD in the treated effluent was predominantly from the slowly degradable fraction, giving inconsistent results. Taking the multi-component nature of the wastewater into account, batch test results can be used to predict fed-batch and continuous activated sludge reactor performance.     

Comparison of hydrolytic enzyme systems in pure culture and activated sludge under different electron acceptor conditions

Enzymatic hydrolysis under different electron acceptor conditions in nutrient removal activated sludge treatment processes is a weak link in the Activated Sludge Model no. 2 (Henze et al., 1995). An experimental study was undertaken to gain insight into the hydrolysis process with specific focus on hydrolysis kinetics and rates under different electron acceptor conditions. Two pure cultures, Bacillus amyloliquefaciens (Gram positive) and Pseudomonas saccharophila (Gram negative) were chosen for the study. In addition, activated sludge grown in an anaerobic-aerobic system was tested for enzymatic activity using starch as the model substrate. The hydrolytic enzymes were found to be released into the bulk in pure cultures whereas the enzyme activity was found to be mainly associated with the cell surfaces in activated sludge. Further, it was observed that the development of the hydrolytic enzyme system in Bacillus amyloliquefaciens and P. saccharophila is strongly suppressed under anoxic and anaerobic conditions. However, the effect of anaerobic and aerobic incubation on hydrolytic enzyme activity in activated sludge was found to be small. Starch hydrolysis kinetic data from batch experiments with activated sludge followed substrate saturation kinetics that were linear with biomass concentration. Finally, the similar hydrolytic enzyme activities observed under anaerobic and aerobic phases of a sequencing batch reactor are explained by considering the aspects of enzyme location and enzyme system development under aerobic and anaerobic phases. It is proposed that the floc bound enzymes are recycled in a single sludge system so that an equilibrium exists between enzyme loss and synthesis at steady state.     

A national survey of activated sludge separation problems in The Czech republic: Filaments,floc characteristics and activated sludge metabolic properties

The extent of activated sludge separation problems associated with excessive growth of filamentous microorganisms has increased recently in the Czech Republic. Most of the activated sludge plants in the country were contacted by mail with a questionnaire, the aim of which was to obtain the basic technological data on the plants and on the nature and intensity of activated sludge separation problems. Selected activated sludge plants were visited at different periods of the year and samples of activated sludge mixed liquor and foam were collected for microscopic examination and kinetic batch tests. It was found that good settling properties were associated more with good quality activated sludge flocs than with the absence of filaments. On the other hand, a very high abundance of filaments always resulted in bulking or foaming. The most common tilamentous microorganisms in activated sludge mixed liquors were M. parvicella, N. limicola and Types 0092,0041 and 0803 while the foams were mostly dominated by M. parvicella, NALOs (GALOs) and N. limicola. Seasonal shifts in filamentous population diversity were also observed. M. parvicella, Type 0092, N. limicola, Type 0803 and Type 0041 were dominant in both conventional and biological nutrient removal activated sludge plants whereas the other Eikelboom's types of filaments dominated mostly in conventional activated sludge plants. The ability of foams to denitrify depended on the dominating tilamentous microorganism and type of substrate.     

Effectiveness of oligonucleotide probes targeted against Thiothrix nivea and type 021n 16s rRNA for in situ identification and population monitoring in activated sludges

In situ hybridization (FISH) of two fluorescent oligonucleotide probes, TNI and 21N, directed against 16S rRNA fragments of Thiothrix nivea and type 021N filamentous bacteria, was used to study activated sludge samples of various origins. Comparison of the results obtained by this technique with morphological data enabled us to evaluate the sensitivity and specificity of the probes, as well as to propose practical classification criteria, in an effort to correlate the identification and ecology of these two microorganisms. We applied the two oligonucleotide probes to the study of Thiothrix sp. in pilot and laboratory batch reactors fed a substrate that was moderately deficient in rapidly available phosphorus. We monitored the growth dynamics of Thiothrix sp. in the pilot reactor, using various hydraulic configurations and after certain perturbations, such as transient substrate overloads. The results provided data which led us to question the importance of the length of time oxygen-deficient sludges spend in clarifier tanks and we discuss the effects of both moderate reductions in oxygen and of very intermittent feedings on the occurrence of Thiothrix sp. growth peaks. The identification and quantification of these filamentous bacteria by size class, made possible by in situ hybridization, are shown to be powerful tools for the early detection and evaluation of episodes of proliferation of filamentous bacteria, revealing them to be much more sensitive indicators than the sludge volume index (SVI).     

活性炭促进UASB处理高浓度啤酒废水效果研究

高浓度啤酒废水在厌氧消化过程中容易酸化,抑制甲烷转化。为提高UASB反应器的稳定性和处理效果,本研究通过序批式试验的方法,以模拟啤酒废水为底物,考察了两种不同粒径的活性炭对加速UASB污泥颗粒化进程和提高甲烷产率的影响。试验运行95 d,进水有机负荷从2.9 kg/(m3·d)增加到12.0 kg/(m3·d),监测反应器内产气量、出水VFA、出水TOC及污泥特性等参数的变化趋势。结果表明:投加活性炭能有效缩短厌氧污泥颗粒化的时间,增强产甲烷菌活性,大幅提升甲烷产量,出水TOC和VFA都维持在较低水平;并且较小粒径的粉末活性炭对UASB反应器的促进作用优于较大粒径的颗粒活性炭,能有效促进底物向甲烷气体转化。     

Evaluation of filamentous microorganism growth factors in an industrial wastewater activated sludge system

An investigation comprising four studies was undertaken to determine possible factors affecting the growth of several different types of filamentous microorganisms present in a bulking industrial wastewater activated sludge. Results from laboratory-scale continuous-flow and full-scale studies suggested that DO concentration and F:M ratio were the likely key factors affecting filamentous growth in the activated sludge. From the results of two laboratory studies isolating the effects of DO concentration and F:M ratio on filamentous growth, favorable growth ranges of DO concentration or F:M ratio were estimated for the following filaments: Microthrix parvicella, Nocardia spp., Nostocoida limicola II, and Types 0041, 1851, and 1863. Most of the bacteria causing filamentous bulking of the activated sludge were found to be filaments typically associated with low F:M, and increasing the F:M ratio appeared to cause N. limicola II to lose its competitive advantage in the activated sludge system. Type 1863, on the other hand, was found to be a low DO filament, as DO concentrations of 0.1 mg O₂/l or less appeared to be a necessary condition for its filamentous growth. Though Nocardia was found to be a low F:M filament, its growth also seemed to be affected by DO concentration, as its growth was stimulated by concentrations of 1.0 mg O₂/l or greater, with a near linear relationship up to at least 5 mg O₂/l.     

Anomalies in the transformation of 3-chlorobenzoate in percolation columns with pseudomonas sp. strain B13

The biotransformation of 3-chlorobenzoate (3CB) by attached cells of Pseudomonas sp. strain B13 was studied in percolation columns operated at various flow rates and biomass contents. Steady-state residual effluent concentrations were compared with predictions from spreadsheet models describing the combined action of microbial and mass transfer kinetics. The effluent concentrations were successfully predicted above a critical ratio of flow rate and biomass. Below this critical point the steady-state residual effluent concentrations were higher than predicted and this deviation increased with decreasing flow rate:biomass ratios. The results are discussed in the light of convection-diffusion processes limiting the transport of both substrate and oxygen to the attached cells.     

In situ characterization of substrate uptake by Microthrix parvicella using microautoradiography

Microthrix parvicella is a filamentous microorganism responsible for bulking and foaming problems in many activated sludge treatment plants. The problems have increased with the introduction of nutrient removal in many countries, and presently, there is no reliable control strategy for M. parvicella. Little is known about the physiology of M. parvicella, and conflicting data exist about its preferred organic substrates, and whether it is able to be physiologically active under anaerobic and anoxic conditions. In this study, the ability of M. parvicella to take up various radioactively labeled organic compounds was investigated in situ at three nutrient removal plants using a microautoradiographic technique. Of 12 compounds tested under aerobic conditions only the long chain fatty acids (LCFA), oleic acid and palmitic acid, and to some extent a lipid, trioleic acid, were assimilated. None of the simple substrates such as acetate, propionate, butyrate, glucose, ethanol, glycine and leucine were taken up. Furthermore, the uptake of oleic acid was compared under anaerobic, anoxic and aerobic conditions, and it was demonstrated that in addition to aerobic conditions M. parvicella was also able to take up oleic acid under anaerobic and anoxic conditions. No difference in substrate uptake pattern for M. parvicella was found among the tested activated sludge plants. The results strongly indicate that a better control strategy against M. parvicella must rely on a better understanding of presence and availability of triglycerides and LCFA, and an improved knowledge of the physiology of M. parvicella under anaerobic and anoxic conditions.     

The use of immersed membranes for upgrading wastewater treatment plants

Membranes can be installed in the clarifier (or aeration tank) of an existing activated sludge plant to enhance the biomass separation function of the system, thereby effectively overcoming any operating constraints associated with sludge settleability. The resulting upgraded plant can be operated at high biomass concentrations (10–20 gMLSS/L), leading to an increase in its treatment capacity. The membranes also ensure a treated water consistently free of suspended solids and a superior disinfection performance. The system offers an enhanced operating flexibility, and allows to operate at high sludge ages leading to a low excess sludge production.Such an immersed membrane activated sludge process (BIOSEP^®) has been developed and applied to the treatment of raw sewage. When treating screened raw sewage with this process, with a sludge concentration of 15 gMLSS/L and a volumetric loading of 1.2 kgCOD/m³/d, a 96% COD reduction and a 95% Total Kjeldahl Nitrogen (TKN) reduction have been obtained. The disinfection performance of the system was over 6 Log removal for fecal coliforms. The resulting production of sludge was 0.20 kgMLSS/kgCOD.Two desk case studies are given for 900 m³/day upgraded plants. In one case, the primary objective was to increase the treatment efficiency and develop nutrient removal for the original plant, while in the other case the primary objective was to increase the capacity of the original 460 m³/day plant.     

Anaerobic processes in activated sludge

We found anoxic zones in aerated activated sludge flocs, and demonstrated denitrification under normal operating conditions. Sulfate reduction was not found. Micro-environments and microbial conversions in flocs from bulking and non-bulking activated sludge were determined with microsensors for H₂S, O₂, NO₂₋ and NO₃₋. Denitriftcation and sulfate reduction rates were measured with ¹⁵N- and ³⁵S-tracer techniques. We showed that under normal reactor conditions (ca. 20% air saturation) anoxic zones develop within flocs allowing denitrification. The denitrtftcation rates amounted to 40% of the rates under anoxic conditions. At 100% air saturation no anoxic zones were found and no denitrification occurred. However, in flocs from bulking sludge (at 20% air saturation) anoxic zones were absent and denitrification did not occur. In bulking sludge only at total anoxia was denitrification found. Confocal microscopy showed that flocs from bulking sludge were much looser than those from non-bulking sludge. The absence of anoxic zones and of denitrification was attributed to the open floc structure, allowing advective oxygen transport.Sulfate reduction was not detected in any of the sludges tested by microsensors or by tracer techniques even under anoxic conditions. This indicates that the sulfur cycle (sulfate reduction and sulfide oxidation) does not play a role in mineralization processes and bulking in activated sludge. Preliminary molecular work (in situ hybridization with the 16S-rRNA probe SRB385) indicated the presence of small amounts of sulfate reducing bacteria in all sludges. Either the probe is not specific or the sulfate reducers present are not active under reactor conditions.     

Variability of type 021n in activated sludge as determined by in situ substrate uptake pattern and in situ hybridization with fluorescent rRNA targeted probes

Bulking sludges were investigated in seven industrial or municipal activated sludge treatment plants from Denmark, Germany and Australia. The dominating filaments were all identified as type 021N according to the Eikelboom key. The extent of variability in the filament taxonomy was assessed using fluorescence in situ hybridization (FISH) with rRNA-targeted nucleic acid probes specific for type 021N, Thiothrix and Leucothrix. Not all of the filaments morphologically identified as type 021N hybridized with the 021N probe. In one treatment plant the predominant filament hybridized with the probe for Thiothrix and in one treatment plant the predominant filament did not hybridize with any of these probes. In none of the plants did filaments hybridize with the probe for Leucothrix. A study of the in situ uptake of different organic substrates by the various filaments was also conducted using microautogradiography. The uptake of 6 different organic substrates under aerobic conditions was studied by providing C-14 or H-3 labeled substrates (acetate, glucose, ethanol, glycine, leucme and oleic acid) in incubations of a period of 3 hours. No filaments took up all the tested substrates, and type 021N from the various treatment plants varied in their uptake abilities. The study demonstrated that strain differences with regard to substrate utilization are likely to occur among bacteria within the same genera and designated types which are indistinguishable on the basis of morphological observations alone and by the molecular probes used in this study for identification. Whether there is a clear correlation between type of wastewater and the capability of taking up the various organic substrates for the filaments remains to be elucidated.     

Characterization of filamentous foaming in activated sludge systems using oligonucleotide hybridization probes and antibody probes

A quantitative method was developed for estimating Gordona mass in activated sludge foam and mixed liquor samples. The technique involves in situ hybridization with a genus-specific fluorescently labeled oligonucleotide probe calibrated on pure cultures of Gordona. The immunofluorescent technique of Hernandez et al. was modified to allow staining with fluorescently labeled antibody and hybridization probes. The results of this technique were compared to those from membrane hybridization studies using radioactively-labeled oligonuelcotide probes. Quantitative membrane hybridizations, in situ hybridizations, and antibody staining resulted in significantly different levels of Gordona in activated sludge foam, activated sludge mixed liquor, return activated sludge, and anaerobic digester sludge. Simultaneous staining with labeled antibodies and oligonucleolide probes provide a definitive identification for Gordona, and represents a new approach for in situ studies of this organism's role to foaming.     

Causes of,and control strategies for,Microthrix parvicella bulking and foaming in nutrient removal activated sludge systems

Bench and pilot scale nutrient removal activated sludge units were used to examine the effect of factors such as temperature, substrate type (easily biodegradable in the form of acetate and slowly biodegradable in the form of oleic acid) on Microthrix parvicella growth. The configurations examined include complete mix with and without selectors (anoxic and anaerobic) and plug flow reactors. The results indicate that low temperatures and substrates in the form of long chain fatty acids favour the growth of M. parvicella. With respect to reactor configuration, a plug flow configuration was shown to be quite effective in controlling the growth of M. parvicella and producing a sludge with good settling characteristics, while the presence of a selector, either anoxic or anaerobic, had no significant effect on the growth of M. parvicella.     

IAL-CHS (internal airlift loop--ceramic honeycomb supports) reactor used for biodegradation of 2,4-dichlorophenol and phenol.

The internal airlift loop reactor with ceramic honeycomb supports (IAL-CHS) was applied for biodegradation of 2,4-dichlorophenol (2,4-DCP) and phenol. A strain of DCP-degrading bacteria isolated from activated sludge, Achromobacter sp., was rapidly immobilized onto the ceramic honeycomb supports. The immobilized cells effectively biodegraded 2,4-DCP alone and together with phenol in batch and continuous-flow experiments. For example, 2,4-DCP was biodegraded from an influent concentration of 50 mg/L to less than 1 mg/L with a 6-h hydraulic retention time (HRT) in continuous flow tests. The immobilized biomass grew and accumulated through 2,4-DCP biodegradation, and the rate of degradation increased accordingly.     

载锰污泥活性炭催化臭氧氧化草酸废水的研究

为研究在催化臭氧化过程中,载锰污泥活性炭对草酸废水降解的催化效果,以及为污泥的资源化探索一条新途径,本实验采用连续流臭氧氧化实验,通过单因素实验方法考察了载锰污泥活性炭催化臭氧化反应的主要影响因素及最佳反应条件,同时对催化臭氧化反应机理进行了探讨。结果表明:在臭氧浓度为5.0 mg/L,载锰污泥活性炭投加量为100 mg/L,pH值为3.5的最佳反应条件下,60 min内草酸的去除率最高达91.2%。叔丁醇对草酸降解具有抑制作用,催化臭氧化反应符合羟基自由基的反应机制。该方法的处理效果较为明显,对评价催化臭氧法的效果具有较明显的参考价值。