Microbial community change of sulfate reduction and sulfur oxidation bacteria in the activated sludge cultivated with acetate and peptone.

The growth of sulfate reducing bacteria (SRB) and filamentous sulfur bacteria was monitored on a laboratory scale in activated sludge reactors using acetate and peptone as the artificial wastewater. When the artificial wastewater contained acetate and peptone, filamentous bacteria increased in the sludge and the SVI values increased. There was a good correlation between sulfate reducing activity and sulfur oxidation activity in the produced sludge. The microbial community change of filamentous sulfur bacteria and sulfate reducing bacteria was analyzed using the fluorescent in situ hybridization (FISH) method. The tendency for the growth of filamentous sulfur bacteria Thiothrix eikelboomii following the growth of SRB was observed. The percentage of SRB385- hybridized cells and DNMA657-hybridized cells found in the total cell area increased from 2-3% to 7-10% when the filamentous bulking occurred.     

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.     

Effect of chlorination bulking control on water quality and phosphate release/uptake in an anaerobic-oxic activated sludge system.

This study evaluates the effect of chlorination bulking control on water quality and phosphate release/uptake in an anaerobic-oxic activated sludge system. A series of batch experiments with different specific NaOCl mass dose were conducted to determine the sludge settling properties, supernatant water quality and phosphate metabolism behavior of filamentous bulking sludge. The harvested sludge was from a continuous-flow anaerobic-oxic (A/O) activated sludge pilot-plant, i.e., enhanced biological phosphorus removal (EBPR) system, operated with 15 days of sludge retention time. The filamentous bacteria in the A/O pilot plant were identified to be Thiothrix according to Eikelboom's classification techniques, which was in accordance with the high influent sulfate concentration of this study (50 mg/L sulfate). Increasing NaOCI concentration, as revealed by experimental results, obviously decreased the sludge settling properties (SVI values and zone settling velocities) and meanwhile significantly reduced supernatant water quality (COD, SS, TP) mainly due to higher suspended solids caused by floc disruption. Moreover, the nine-hour batch experiments indicated that high NaOCI dosage (40 mg/gMLSS) completely deteriorated phosphate metabolism of EBPR sludge. Such a high dosage of chlorination further confirmed overdosing through disappearance of intracellular PHB and death of protozoa by microscopic investigation. Still, phosphate release/uptake behavior of EBPR sludge properly functions at low NaOCl dosage (5 mg/g MLSS). Besides, phosphate metabolism worsens rapidly before the SVI value reaches its lowest level. These findings imply that determining NaOCI requirement with merely SVI values can readily result in chlorination overdosing. Proper NaOCI dosage requires a delicately balanced consideration between sludge settling improvement, water quality demand and phosphate metabolism. Batch test of phosphate release/uptake is apparently a prerequisite to conclude an appropriate NaOCl dosage for bulking control.     

基于不同碳源的硫酸盐还原菌处理硫酸盐酸性废水试验

为探究硫酸盐还原菌(sulfate reducing bacteria,SRB)在不同碳源条件下还原硫酸盐的最佳pH值条件及其还原动力学过程,分别以甘蔗渣和乳酸钠为碳源,负载SRB处理含硫酸盐酸性废水。结果表明:以甘蔗渣、乳酸钠为碳源,SRB均在pH为6时对硫酸盐的去除效果最佳,最大去除率分别为71. 82%、85. 31%,体系氧化还原电位分别为-242、-164 m V;而在pH为4条件下,SRB对硫酸盐的去除效果甘蔗渣优于以乳酸钠为碳源的体系,对硫酸盐最大去除率分别为49. 04%、36. 24%,体系氧化还原电位分别为-229、-57 mV;两种碳源条件下,SRB还原SO_4~(2-)的过程都符合一级动力学模型,在以甘蔗渣为碳源、pH为6的体系中,其最大还原速率分别为0. 20007/d、0. 12688/d。     

污泥膨胀的原因分析及预警监测研究

通过分析活性污泥法污水处理系统的实际运行工况,筛选出丝状细菌数量级、污泥沉降比、负荷比、溶解氧、泥龄、温度和营养物类型作为污泥膨胀指标,并根据研究得出污泥膨胀监控指示范围.     

Occurrence and control of filamentous bulking in aerated wastewater treatment plants of the French paper industry.

The occurrence of filamentous bacteria was investigated in 15 French pulp and paper activated sludge wastewater treatment plant (WWTP). Large filamentous populations were present in most of the plants. Identification carried out with conventional methods based on morphological features and staining techniques showed that the four main filamentous bacteria encountered in these industrial WWTP and responsible for bulking belong to the genera Thiothrix sp., Type 021 N, Haliscomenobacter hydrossis and Type 0092. During two years a specific survey was performed for three of these WWTP showing recurrent bulking phenomena. Data from WWTP performance, chemical data and filaments characterization were compared to correlate the presence of specific filaments with process operating conditions.     

污泥膨胀的原因分析及预警监测研究

通过分析活性污泥法污水处理系统的实际运行工况,筛选出丝状细菌数量级、污泥沉降比、负荷比、溶解氧、泥龄、温度和营养物类型作为污泥膨胀指标,并根据研究得出污泥膨胀监控指示范围。     

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.     

氧化沟工艺污泥膨胀及出水水质影响因素的研究

针对丝状菌污泥膨胀造成改良式氧化沟工艺处理城市生活污水超标的问题,通过分析进水水质、溶解氧、温度、污泥膨胀指数、出水水质变化的关系,探讨了导致丝状菌膨胀的主要限制因子以及出水水质的变化.研究结果表明,此工艺中进水BOD5、CODcr、TP浓度和pH值变化不是导致污泥膨胀的原因,进水TN和环境温度对污泥膨胀略有影响,DO、NH3-N变化与SVI有较强的相关性,DO和NH3-N越高,SVI越低.     

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.     

Identity, abundance and physiology of Aquaspirillum-related filamentous bacteria in activated sludge.

Microcolony-forming bacteria closely related to the genus Aquaspirillum in the Betaproteobacteria were recently observed to be abundant in many nutrient removal wastewater treatment plants. The developed oligonucleotide probe, Aqs997, however, occasionally also targeted some filamentous bacteria in activated sludge samples when fluorescence in situ hybridization was performed. In this study, the identity, abundance, and ecophysiology of these Aqs997-positive filamentous bacteria were studied in detail. Most of the Aqs997-positive filamentous bacteria could morphologically be identified as either Eikelboom Type 1701, Type 0041/0675 or possibly Type 1851, all characterized by epiphytic growth. They were found in almost all 21 wastewater treatment plants investigated. Two morphotypes were found. Type A filaments, which seemed to be the same genotype as the microcolony-forming bacteria targeted by probe Aqs997.Type B filaments also hybridized with probe GNS941, specific for the Chloroflexi phylum, so the true identity remains unclear. Aqs997-positive filaments usually stained Gram-negative, but Gram-positive filaments were also found, stressing the difficulties in identifying bacteria from morphology and simple staining results. Studies on the ecophysiology by microautoradiography showed that Aqs997-positive filamentous bacteria did not consume acetate and glucose, while some took up butyrate, mannose, and certain amino acids. Most likely, some Aqs997-positive filamentous bacteria were able to perform full denitrification such as the Aqs997-positive microcolony-forming bacteria, and some were able to store polyhydroxyalkanoates under anaerobic conditions, potentially being glycogen accumulating organisms.     

城市污水处理厂的污泥膨胀研究分析

通过对北京市大型城市污水处理厂优势丝状菌鉴定及温度、生物选择器和溶解氧对污泥膨胀影响的研究分析,发现脱氮除磷工艺普遍会在冬季发生由Microthrix parvicella茵引起的污泥膨胀.研究了温度与SVI的变化规律,分析了出水亚硝酸氮和SVI的相关性,并对生物选择器和不同DO浓度对污泥膨胀的影响进行了分析.     

阿哈湖和洱海沉积物硫酸盐还原菌研究

采用MPN法分析了贵州阿哈湖和云南洱海春秋两个季节沉积物硫酸盐还原菌含量,结果表明淡水湖泊沉积物硫酸盐还原菌含量低于海洋沉积物;洱海沉积物硫酸盐还原菌含量低于阿哈湖沉积物,秋季沉积物硫酸盐还原菌含量高于春季,且有该类微生物栖息的沉积物深度增加。分析造成硫酸盐还原菌含量发生改变的环境因素可能是沉积物中硫酸根浓度、温度以及有机质的变化。     

Effect of chemical and biological factors on the densification of filamentous sludge

Densification may be an alternative method for controlling sludge bulking; however, little information is available on the method. Various methods including different oxygen concentrations, surface velocities of aeration, calcium concentrations and extended starvation period were employed for triggering densification of filamentous sludge in this study. It was found that high surface velocity of aeration could effectively improve the settleability of filamentous sludge in the short term, which resulted in filamentous granulation, but could not avoid filamentous bulking even after granulation. Increase of calcium concentration could also improve the settleability by forming pigtail filaments colony in the short term, but could not trigger granulation. Extension of starvation period seems effective neither for further densification nor for granulation. Additionally, increased ovality of meshes formed by filaments was frequently associated with a dense structure of filamentous sludge. It was suggested that pigtail type of filamentous colony that thus increased the resistance to elevated shear force may encourage the densification of sludge, and two kind of densification growth patterns, i.e., granular or cluster growth pattern of filamentous sludge, were identified.     

The in situ physiology of "Nostocoida limicola" II, a filamentous bacterial morphotype in bulking activated sludge, using fluorescence in situ hybridization and microautoradiography.

The in situ physiology of the actinobacterial bulking and foaming filamentous bacterium "Nostocoida limicola" II was studied by fluorescence in situ hybridization/microautoradiography. Substrate assimilation patterns of pure cultures of this bacterium were different to those seen in activated sludge biomass samples. There was no evidence to suggest that "N. limicola" II preferred hydrophobic substrates, but evidence was produced to support the view that it is metabolically active under anaerobic conditions in activated sludge.     

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.     

硫酸盐还原菌及其在废水厌氧治理中的应用

硫酸盐还原菌(SRB)在废水处理方面有独特的优势,在厌氧环境中能以硫酸盐作为电子受体降解有机污染物。本文阐明了SRB处理废水中污染物的机理,综述了国内外利用SRB处理重金属离子废水、含硫酸盐有机废水和酸性矿山废水的研究进展。最后总结了目前在工程应用方面尚存在的问题。     

Development of single-stage nitrogen removal using anammox and partial nitritation (SNAP) and its treatment performances.

Single-stage Nitrogen removal using Anammox and Partial nitritation (SNAP) process was newly developed as an economical nitrogen removal process for ammonium rich wastewaters. The experimental studies for the evaluation of SNAP process were carried out using a novel biofilm reactor, in which hydrophilic net-type acryl fiber biomass carrier was applied. This SNAP reactor was operated under operational conditions of pH 7.5-7.7, 35 degrees C and DO 2-3 mg/L, and 60 to 80% of influent NH4-N was removed under loading rate of 0.48 kg-N/m3/d. Through the DNA analysis of the attached sludge, it was made clear that ammonium oxidizing bacteria (AOB) and anammox bacteria coexisted in the attach-immobilized sludge on the acryl fiber biomass carrier. Favorable conditions for the growth of anammox bacteria were created inside attach-immobilized nitrifying sludge. Two kinds of anammox bacteria and two kinds of AOB were detected in the SNAP sludge. Existence ratios of anammox and AOB were estimated to be 15% and 8.7%, respectively, based on the obtained clone numbers. This coexisting condition was confirmed by the FISH image of SNAP sludge and its confocal laser scanning microscope.     

Modification of ASM3 for the determination of biomass adsorption/storage capacity in bulking sludge control.

The selector activated sludge (SAS) systems are known to prevent excessive growth of filamentous microorganisms responsible for bulking sludge, but these systems were hardly ever modelled. This study aimed to develop a model capable of predicting rapid substrate removal in the SAS systems. For this purpose, the Activated Sludge Model No. 3 (ASM3) was extended with three processes (adsorption, direct growth on the adsorbed substrate under aerobic or anoxic conditions). The modified ASM3 was tested against the results of batch experiments with the biomass originating from two full-scale SAS systems in Germany. The endogenous biomass was mixed with various readily biodegradable substrates (acetate, peptone, glucose and wastewater) and the utilisation of substrate (expresses as COD) and oxygen uptake rates (OURs) were measured during the experiments. In general, model predictions fitted to the experimental data, but a considerable number of kinetic (5) and stoichiometric (2) parameters needed to be adjusted during model calibration. The simulation results revealed that storage was generally a dominating process compared to direct growth in terms of the adsorbed substrate utilisation. The contribution of storage ranged from 65-71% (Plant A) and 69-92% (Plant B).