Impact of septic compounds and operational conditions on the microbiology of an activated sludge system.

In activated sludge (AS) biotreatment, septic compounds such as volatile organic acids and reduced sulphur compounds have been frequently cited as a major cause of Thiothrix and Type 021N filamentous bulking. These filaments are common in Canadian pulp and paper biotreatment systems, where they cause settling problems in secondary clarifiers. We conducted a 14-week study of a TMP/newsprint mill effluent to characterize the septic compounds entering the biotreatment, and to determine correlations with AS biomass characteristics and biotreatment operating parameters. A significant correlation was found between the sludge volume index, the abundance of Type 021N, and the propionic acid (PA) concentration in the primary clarified effluent. PA also induced a significant change in the flocculating bacteria size distribution determined by digital imaging. Consequently, the correlation observed between PA and Type 021N bulking is an indirect effect of inhibition of floc-forming microorganisms, giving a competitive advantage to filaments.     

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.     

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.     

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.     

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).     

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.     

Nitrogen removal and sludge reduction in a symbiotic activated sludge system between anaerobic archaea and bacteria.

The possible symbiosis between bacteria and anaerobic archaea was investigated in intermittent aeration (I/A) systems. Archaea solution added to I/A reactor might play an important role in biological activities as well as in improvement of mineralization of organic matter. I/A reactor with archaea solution (I/A-arch) could increase both nitrification and denitrification rate and also reduce the sludge yield remarkably. These results indicate the possibility of the symbiotic activated sludge system with anaerobic archaea by controlling the DO level in the aeration tank. In this study, DO was controlled by intermittent aeration schemes and a successful symbiotic activated sludge system was achieved to reach the following conclusions. 1) SOUR of I/A-arch system was 2.9 mg-O2/g-VSS x min. SOUR and nitrification rate of the sludge from I/A-arch was higher than those from the I/A and A/S reactors. 2) Removal efficiencies of organic matter (TCOD(Cr)) in I/A-arch, I/A and conventional activated sludge (A/S) reactors were 93, 90 and 87%, respectively. 3) Nitrification occurred successfully in each reactor, while denitrification rate was much higher in the I/A-arch reactor. Efficiencies of TN removal in A/I-arch, I/A and A/S reactors were 75, 63 and 33%, respectively. 4) Observed yield coefficients of I/A-arch, I/A and A/S reactors were 0.28, 0.41 and 0.37 g-VSS/g-COD.     

Control of bulking in a full scale plant by addition of talc (PE 8418)

A very severe bulking problem (Type 021 N) in the water treatment plant of Bodegraven in The Netherlands was solved by the addition of a specially shaped talqueous powder, PE 8418. After addition of this mineral product, the SVI dropped almost instantaneously from 850 ml/g to 250 ml/g and over the next 2 weeks decreased further to 100 to 125 ml/g. A ratio PE 8418/biomass of 0.6:1 was sufficient to keep the SVI at this level. The settling velocity of the sludge increased from <1.0m/h to 1.5 m/h. These settling properties allowed normal plant operation. During the first 2 weeks of the experiment, scum on the final clarifier and the aeration tank caused some problems. It is assumed that the complete incorporation of PE 8418 particles into the flocs takes some time, if a large quantity of this product is added at once. Meanwhile, free particles may contribute to stabilization of gas bubbles, viz. flotation of activated sludge.     

Interactions between filamentous sulfur bacteria,sulfate reducing bacteria and poly-P accumulating bacteria in anaerobic-oxic activated sludge from a municipal plant

The interactions between filamentous sulfur bacteria (FSB), sulfate reducing bacteria (SRB) and poly-P accumulating bacteria (PAB) in the activated sludge of a municipal plant operated under anaerobic-oxic conditions were examined in batch experiments using return sludge (RAS) and settled sewage. Phosphate release and sulfate reduction occurred simultaneously under anaerobic conditions. SRB were more sensitive to temperature changes than PAB. SRB played an important role in the decomposition of propionate to acetate. When the sulfate reduction rates were high, there was a tendency for the maximum release of phosphate also to be high. This was explained by the fact that PAB utilized the acetate produced by SRB. Sulfur oxidizing bacteria were sensitive to temperature change. When the sulfate reduction rate was high, the sulfide oxidizing rate was also high and filamentous bulking occurred. The results showed that sulfate reduction was a cause of filamentous bulking due to Type 021N that could utilize reduced sulfur.     

Monitoring activated sludge settling properties using image analysis.

The goal of this study is to develop a monitoring system for activated sludge properties, as this is an essential tool in the battle against filamentous bulking. A fully automatic image analysis procedure for recognising and characterising flocs and filaments in activated sludge images has been optimised and subsequently used to monitor activated sludge properties in a lab-scale installation. The results of two experiments indicate that the image information correlates well with the Sludge Volume Index. It is shown that, at the onset of filamentous bulking, there is an increase in total filament length on the one hand, and a change in floc shape on the other hand.     

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.     

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.     

Aeration tank odour by dimethyl sulphoxide (DMSO) waste in sewage.

Sewage plants can experience dimethyl sulphide (DMS) odour problems by at least one mg/L dimethylsulphoxide (DMSO) waste residue in plant influent, through a DMSO/DMS reduction mechanism. This bench-scale batch study simulates in bottles the role of poor aeration in wastewater treatment on the DMSO/DMS and sulphate/H2S reduction. The study compares headspace concentrations of sulphide odorants developed by activated sludge (closed bottles, half full) after six hours under anoxic versus anaerobic conditions, with 0 versus 2 mg/L DMSO addition. Anoxic sludge (0.1 - 2 mg/L dissolved oxygen, DO) with DMSO resulted in about 50 ppmv DMS and no other sulphide, while DMSO-free sludge was free of detectable sulphides. Anaerobic sludge (no measurable DO to the point of sulphate reduction) with DMSO resulted in 22/4/37 ppmv of H2S/methanethiol (MT)/DMS, while DMSO-free sludge resulted in 44/8/2 ppmv of H2S/MT/DMS. It is concluded that common "anoxic" aeration tank zones with measurable DO in bulk water but immeasurable DO inside sludge flocs (nitrate reducing) experience DMSO reduction to DMS that is oxidation resistant and becomes the most important odorant. Under anaerobic conditions, H2S from sulphate reduction becomes an additional important odorant. A strategy is developed that allows operators to determine from the quantity of different sulphides whether the DMSO/DMS mechanism is important at their wastewater plant.     

Treatment of municipal wastewater in a hybrid process using a new suspended carrier with large surface area.

An activated sludge/biofilm hybrid process treating municipal wastewater was studied in pilot plant trials. A new type of suspended carrier, with large effective surface area, was employed in the process with the aim of enhancing nitrification. The pilot plant was operated for 1.5 years in five different configurations including pre-denitrification in all five and enhanced biological phosphorus removal in the final two. The wastewater temperature ranged between 11 degrees C and 20 degrees C, and the nominal dissolved oxygen (DO) level was 5-6 mg/L. The nitrification rate obtained on the new carrier within the hybrid stage was in the range of 0.9-1.2 g NH4-N/m2/d corresponding to a volumetric rate of 19-23 g NH4-N/m3/h (total nitrification including nitrification in the suspended solids). More than 80% of the total nitrification took place on the carrier (and the remainder in the suspended solids). The nitrification rate was shown to correlate with DO, decreasing when the DO was decreased. The results supported the idea of using the new carrier as a tool to upgrade plants not having nitrification today or improve nitrification in activated sludge processes not reaching necessary discharge levels. The large surface area present for nitrification makes it possible to obtain high nitrification rates within limited volumes. The possibility to keep the total suspended solid content low (< 3 g/L) and avoiding problems with the filament Microthrix parvicella, are other beneficial properties of the hybrid process.     

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。     

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.     

Membrane bioreactor (MBR) sludge inoculation in a hybrid process scheme concept to assist overloaded conventional activated sludge (CAS) process operations

This study analyzes the effect of inoculating membrane bioreactor (MBR) sludge in a parallel-operated overloaded conventional activated sludge (CAS) system. Modelling studies that showed the beneficial effect of this inoculation were confirmed though full scale tests. Total nitrogen (TN) removal in the CAS increased and higher nitrate formation rates were achieved. During MBR sludge inoculation, the TN removal in the CAS was proven to be dependent on MBR sludge loading. Special attention was given to the effect of inoculation on sludge quality. The MBR flocs, grown without selection pressure, were clearly distinct from the more compact flocs in the CAS system and also contained more filamentous bacteria. After inoculation the MBR flocs did not evolve into good-settling compact flocs, resulting in a decreasing sludge quality. During high flow conditions the effluent CAS contained more suspended solids. Sludge volume index, however, did not increase. Laboratory tests were held to determine the threshold volume of MBR sludge to be seeded into the CAS reactor. Above 16-30%, supernatant turbidity and scum formation increased markedly.     

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.