Type 021N Research Articles

Bulking of activated sludge in the biological treatment of municipal and industrial wastewater, due to the massive development of filamentous bacteria Type 021N

A research of the causes and consequences of bulking of activated sludge was conducted at existing biological wastewater treatment plants (municipal and local at an industrial enterprise). The filamentous bacterium that was caused the bulking of the activated sludge in the studied treatment plants was identified as Type 021N. It was established that the studied filamentous bulking causes a significant deterioration of the technological characteristics of activated sludge flocs and it is accompanied by suppression of the nitrification process. To suppress the bulking of activated sludge two methods of suppressing the inflow of hydrogen sulfide into biological treatment plants were successfully used: blowing from wastewater at municipal treatment plants and inhibiting its formation at local treatment plants in the settling and laminar flow zones.

Landfill leachate treatment with a full-scale membrane bioreactor: impact of leachate characteristics on filamentous bacteria.

Bulking and foaming are extreme filamentous bacterial growths that present serious challenges for the biological leachate treatment process. The current study evaluates the performance of long-term full-scale membrane bioreactor (MBR) treating landfill leachate, specifically focusing on filamentous bacteria overgrowth in the bioreactors. The influence of the variation in leachate structure and operational conditions on floc morphology and filamentous bacteria overgrowth were analyzed for 11months of operation of the full-scale MBR system. The average chemical oxygen demand (COD) and NH4-N removal efficiencies of the system were 87.8 ± 4% and 99.5 ± 0.7%. However, incomplete denitrification was observed when the F/M ratio was low. The high C/N ratio was observed to enhance the frequency of small flocs. Furthermore, a poor to medium diversity of the microbial community was observed. Haliscomenobacter hydrossis, Microthrix parvicella, and Type 021N were found as the most numerous filamentous organisms. Paramecium spp., Euplotes spp., and Aspidisca spp. were found in small quantities. The limited concentration of PO4-P in the leachate compared to high COD and NH4-N concentrations most probably caused phosphate deprivation and increased abundance of identified filamentous microorganisms. This work is the first study in Türkiye that investigates the bulking and foaming problem in full-scale MBR that treats landfill leachate. Hence, it may provide some pioneering perspectives into landfill leachate remediation by monitoring the hybrid biological system.

ОСОБЛИВОСТІ СПУХАННЯ АКТИВНОГО МУЛУ В МЕМБРАННИХ БІОРЕАКТОРАХ

Membrane bioreactor (MBR) technology, the practical application of which is growing steadily, is a combination of classical technology of biological treatment with activated sludge with a membrane system of micro- or ultrafiltration. MBR are an alternative to the widely used method of separating activated sludge from wastewater in secondary settling tanks used in traditional biological treatment systems. MBR exclude large-scale negative consequences, as in system with traditional aeration tanks and secondary settling tanks in the "swelling of activated sludge". But "swelling of activated sludge" can affect the nitrification process and minimize the efficiency of ammonium nitrogen removal. The purpose of the work is to study the swelling of activated sludge swelling on the existing membrane bioreactor. The research was carried out by hydrochemical analysis of wastewater in MBR and determination of technological properties of activated sludge in the bioreactor. The negative effect of activated sludge swelling in MBR on nitrification activity in the system was established, which caused a decrease in the efficiency of ammonium nitrogen removal. Filamentous bacteria were identified according to the classification of D. Eickelbum, which caused swelling of activated sludge in the studied MBR, Type 021N.

Microscopic Analysis of Activated Sludge in Industrial Textile Wastewater Treatment Plant

Abstract The relationship between a quality of activated sludge microbiota and wastewater treatment plant (WWTP) operational stability has been defined in the past few decades. However, this dependence is not so clear in the case of industrial wastewater treatment. In this article, a very specific example of industrial textile wastewater treatment plant (ITWTP) is analyzed. Textile effluents are well known as highly contaminated wastewater containing many biodegradable compounds. Microscopic analysis included flocs morphology examination, attempts to evaluate the Sludge Biotic Index (SBI), and identification of dominant filamentous microorganisms. Routine operational control of ITWTP covered pH, temperature, redox potential, dissolved oxygen and COD measurements. The average ecosystem existing in the described ITWTP differed significantly compared to municipal WWTPs. The flocs were smaller and irregular. Filamentous bacteria did not cause foaming although filaments index reached 4. Nostocoida limicola I dominated with significant amounts of type 0041 and type 021N. The evaluation of SBI was impossible as the most of protozoan was in the form of cysts. The overall microbiota diversity correlated with COD removal in activated sludge unit of ITWTP.

Filamentous organisms degrade oxygen transfer efficiency by increasing mixed liquor apparent viscosity: Mechanistic understanding and experimental verification

Recent findings have demonstrated that activated sludge morphology significantly impacts oxygen transfer efficiency (OTE) in the activated sludge process. In this study, we developed a mechanistic understanding of this impact. Mixed liquor samples collected from a domestic wastewater treatment plant (WWTP) were blended with a bulking activated sludge from a bench scale reactor (BSR) cultured on synthetic wastewater to manipulate various morphological parameters such as the settled sludge volume (SV), the sludge volume index (SVI), and the specific filament length (SFL). The filaments that were present in the blended sludges consisted largely of Type 0041 and Type 021N, which are commonly found in WWTPs that treat domestic wastewater. Variations in sludge morphology, as quantified by settled sludge volume after 30 min (SV30), SVI, and SFL, systematically affected the mixed liquor apparent viscosity (μapp), which consequently impacted OTE. An increase in the SFL from 9.61 × 106 μm g−1 to 6.88 × 107 μm g−1 resulted in a 41.4% increase in apparent viscosity and a 24.6% decrease in volumetric mass transfer coefficient (KLa). A new parameter, named the ultimate settleability (SVULT), was developed by curve fitting the SV versus time data and found to relate with μapp through an expanded form of the Einstein Equation for the viscosity. Therefore, SVULT is a corollary for the particle volume fraction that incorporates effects of both the sludge morphology and mass concentration on μapp. Theoretical derivation revealed that an increase in SVULT resulted in an increase in μapp, which reduced oxygen transfer by increasing the air bubble size and reducing refreshment of the liquid at the gas-liquid interface. The KLa was found to be inversely proportional to μapp0.75 through fitting the experimental data with the theoretical model. Using a variance-based global sensitivity analysis, three operating parameters that have the most impact on oxygen transfer were identified: the power input per unit volume, the superficial gas flowrate, and the μapp.

Filamentous bacteria in the nitrifying activated sludge.

Filamentous bacteria in addition to wastewater treatment are responsible for the shape of flocs and sedimentation properties of activated sludge. Their dynamics in activated sludge influences the performance of the whole sewage treatment plant. Therefore the composition of activated sludge biocenosis and its dynamics in the nitrification process were investigated. Four laboratory-scale activated sludge membrane bioreactors fed with wastewater highly concentrated with ammonium (synthetic wastewater imitating landfill leachate) were operated to obtain a high rate of nitrification. The sludge age was 8, 12, 24 and 32 days. An additional fifth reactor was conventionally ammonium loaded at 12-day sludge age and served as the reference. A shift in filamentous bacteria population was observed in all operated reactors. There was no influence of sludge age on composition or abundance of filamentous biocenosis. In high ammonium loaded activated sludge Nostocoida limicola, Haliscomenobacter hydrossis and also Type 021N were the most abundant filamentous bacteria. In the reference reactor Type 021N and Sphaerotilus natans dominated the activated sludge.

Dolomite Reactor, a Retrofitting Approach for Activated Sludge Against Bulking

This study aims to assess the application of natural dolomite for upgrading activated sludge systems that encounter sludge bulking. For this purpose, a pilot of conventional activated sludge with 4.8 cubic meters volume was operated for 10 months in real condition and inoculated intermittently with sludge having filamentous bulking problem. The experimental results showed that dolomite could retain TSS, BOD, and COD removal efficiencies on 90, 95 and 96 %, respectively. It could also reduce the variations of sludge volume and effluent quality. By increasing the dolomite concentrations from 25 g/m3 to the optimum level of 100 g/m3, the soluble calcium was developed 24 %. This could enhance the sludge settleability in which the counts of filamentous bacteria, colony size, and sludge volume index (SVI) reduced 44 %, 1 mm, and 150 mL/g, respectively. Here, Sphaerotilus natans and Type 021N were considerably removed in high dosages. Nevertheless, Microthrix Parvicella showed more resistance. Regarding artificial neural network simulation, it was realized that the optimum dosage of dolomite should be adjusted with MLSS concentrations to keep SVI at the proper range. Consequently, dolomite was verified as efficient retrofitting approach for activated sludge systems. However, this solution requires further studies in full-scale operation.

Evaluation of granular sludge for secondary treatment of saline municipal sewage

This study examined the impact of chemical oxygen demand (COD) loading and dissolved oxygen (DO) concentration on the stability and performance of granular sludge treating high saline municipal sewage. Under high DO concentrations of 4.0–7.0 mg/L, and COD loading rates of 0.98 and 1.55 kg/m3/d, rapid settling granules were established within four weeks of start-up. Under the highest COD load, a reduction in DO lead to the rapid deterioration of the sludge volume index (SVI) and washout of granules due to prolific growth of the filament Thiothrix Type 021N. Conversely, when operated under a lower COD load, a reduction in DO concentration had no adverse impact on the stability of SVI and granules. A decrease in DO also improved nitrogen removal performance, where simultaneous removal of ammonium (98%), total nitrogen (86%) and BOD5 (98%) were achieved when median DO concentrations were between 1.0 and 1.5 mg/L. Phosphate removal was lower than expected, however the level of biological phosphate removal activity observed appeared sufficient to maintain granule stability, even under low DO concentrations. Nitrous oxide emissions were also characterised, which ranged between 2.3 and 6.8% of the total nitrogen load. Our results confirmed that granular sludge is a viable option for the treatment of saline sewage.

Experimental attempt at using Lecane inermis rotifers to control filamentous bacteria Eikelboom Type 0092 in activated sludge.

In laboratory experiments, the authors tested the efficacy of Lecane inermis rotifers in reducing the abundance of filamentous bacteria in activated sludge samples dominated by one filamentous bacterium, Eikelboom Type 0092. The experiment was conducted in three replicates and repeated three times at 1-month intervals. In all repetitions, rotifers were able to significantly reduce the density of bacterial filaments through predation. It was also shown that the influence of rotifers on filament abundance depends on the duration of grazing pressure. The data show that L. inermis, already reported to be effective in reducing the density of Microthrix parvicella, Nostocoida limicola, and Type 021N in activated sludge, can be equally helpful in limiting Type 0092, another troublesome filamentous bacterium found in low-loaded water resource recovery facilities.

Filament identification and dominance of Eikelboom Type 0092 in activated sludge from wastewater treatment facilities in Cape Town, South Africa

Routine characterisation of activated sludge and identification of the filamentous population by microscopic and/or other non-culture dependent techniques can assist in diagnosing the aetiology of poor performance of wastewater treatment works (WWTWs). In South Africa, most facilities rely solely on physicochemical indicators, treating reactors as ‘black-boxes’, with the result that process adjustments are often delayed, to the detriment of the environment. This study was performed in order to gain insight into the filamentous population found in activated sludge in Cape Town WWTWs, to compare these with other global and local literature findings, and to build capacity in this science. Physicochemical and plant performance parameters, in terms of nutrient removal and settling, were obtained from routine operational data and assessed in conjunction with the microscopic analyses of activated sludge samples taken over a 6-month period. Hypotheses on the links between filament types and/or plant configurations and/or operational parameters were formulated using existing literature. In order of prevalence, the five most common dominant filament species in 96 activated sludge samples were: Eikelboom Type 0092, Eikelboom Type 1851, nocardioforms, Microthrix parvicella and Eikelboom Type 021N. In order to compile a statistically significant database, it is recommended that an extensive nationwide study is conducted to link filament types with plant configurations, operational parameters and geographical locations. Keywords : activated sludge, bulking, identification, filament, Type 0092

Effects of feeding pattern and dissolved oxygen concentration on microbial morphology and community structure: The competition between floc-forming bacteria and filamentous bacteria

Understanding of the competition between floc-formers and filaments is critical to prevent filamentous bulking in practice. This study aimed to investigate the effects of feeding pattern and dissolved oxygen (DO) concentration on their competition in four sequencing batch reactors (SBRs). Short feeding under anoxic condition (fill time<10min) resulted in a well-settling sludge (sludge volume index (SVI)<100mL/g), in spite of DO concentrations. Sludge settleability deteriorated (SVI>200mL/g) and filamentous bulking was observed when the substrate was added in a limiting rate by prolonging the anoxic fill time up to 90min. In contrast, sludge settleability in fully aerobic systems was quite poor (SVI>500mL/g) in spite of the feeding length. Compared to the systems with an anoxic fill phase, more types and abundant filamentous bacteria were identified in fully aerobic systems. Microscopic observation, staining reactions and fluorescence in situ hybridization analysis indicated that the extensive filaments, including Thiothrix nivea, Type 021N, Type 1851 and Microthrix parvicella, proliferated in fully aerobic systems. The results of this study indicated that substrate gradients played an important role on the competition between filaments and floc-formers. It is recommend the adoption of plug-flow selector configurations, with anoxic conditions in order to maintain good and robust sludge settleability.

Use of talc for filamentous bulking control in effluent treatment plant

One of the main existing operational problems in activated sludge effluent treatment processes is the poor sedimentation of the biological sludge in the secondary clarifiers. This poor settleability of the sludge is, generally, associated with excessive growth of filamentous bacteria in the bioreactor causing sludge filamentous bulking. In Brazil, many pulp and paper mills have faced problems related to sludge filamentous bulking in their effluent treatment plants experiencing a significant reduction in COD and BOD removal efficiency and a loss of suspended solids in the treated effluent. The addition of talc to increase floc density for the control of filamentous bulking has been studied. The objective of this study was to test the use of talc for the control of filamentous bulking. The bio-sludge used was obtained from a recycling paper mill and had an abundance of filamentous bacteria Type 021N. The experiment consisted of five bioreactors operating batch wise with sludge age of ten days. Different talc applications were used: 0%, 25%, 50%, 75% and 100% in relation to the total suspended solids in the aeration tank. The following analyses were carried out to monitor the system: suspended solids, sludge volumetric index (SVI) and chemical oxygen demand. Successive additions of talc contributed for the reduction of IVL. Dosages of 75% and 100% showed to be most efficient to improve settleability of the sludge. The presence of talc did not interfere with biological activity of the sludge. However, it was observed an increase of the total suspended solid concentration in the aeration tank. The addition of talc had a temporary effect on sludge settleability, so it was necessary to add repeatedly talc in the reactor in order to maintain a low sludge volumetric index.

Filamentous sludge bulking control by nano zero-valent iron in activated sludge treatment systems.

Sludge bulking causes loss of biomass in the effluent and deterioration of effluent water quality. This study explored the use of nano zero-valent iron (NZVI with an average particle size of 55 ± 11 nm) for sludge bulking control. In two Modified Ludzack-Ettinger (MLE) activated sludge treatment systems, a single dose of NZVI at the final concentration of 100 mg Fe per L in the mixed liquor reduced the number of filamentous bacteria Type 021N by 2-3 log units (a reduction of 99.9 and 96.7% in MLE tank #1 and #2, respectively). The side effect of the use of NZVI depended on sludge bulking conditions and biomass concentration. In the system with sludge bulking and significant sludge loss (average biomass concentration of 1022 ± 159 COD mg per L or at the ratio of 0.098 g Fe per g biomass COD), the use of NZVI increased effluent COD, NH4(+)-N and NO2(-)-N concentrations, as also evident with the loss of nitrifying populations and nitrifying activities resulting in more than 40 days to have the full recovery of the activated sludge system. In contrast, in the system with the early stages of bulking and the biomass concentration of 1799 ± 113 COD mg per L (at the ratio of 0.056 g Fe per g biomass COD), the effluent water quality and overall bioreactor performance were only slightly affected for a few days.

The use of rotifers for limiting filamentous bacteria Type 021N, a bacteria causing activated sludge bulking

The excessive growth of filamentous bacteria and the resultant bulking of activated sludge constitute a serious problem in numerous wastewater treatment plants. Lecane inermis rotifers were previously shown to be capable of reducing the abundance of Microthrix parvicella and Nostocoida limicola in activated sludge. In the present study, the effectiveness of four Lecane clones in reducing the abundance of Type 021N filamentous bacteria was investigated. Three independent experiments were carried out on activated sludge from three different treatment plants. We found that Lecane rotifers are efficient consumers of Type 021N filaments.

Control filamentous bulking caused by chlorine-resistant Type 021N bacteria through adding a biocide CTAB

Filamentous bulking sludge due to excessive growth of filamentous bacteria is a serious operational problem in activated sludge plants. The addition of chemicals is one of widespread ways to control filamentous bulking. In this study, filamentous bulking in a continuous activated sludge system was found to be mainly caused by Eikelboom Type 021N filamentous bacteria likely due to low substrate concentration gradients. These Type 021N bacteria were found to be resistant to chlorination, maintaining cell integrity at a dosage of up to 80 mg Cl/gSS. An alternative biocidal agent, cetyltrimethyl ammonium bromide (CTAB), exhibited a much stronger biocidal effect on these filaments, which significantly improved sludge settleability. Type 021N with filamentous index of 5 was selectively killed, but floc-formers recovery their activity after CTAB termination. The study implied that CTAB might have more penetration capacity to cell wall of chlorine-resistant Type 021N bacteria. We therefore suggest the penetration property of filament cell wall should be considered or tested before the selection of biocide type in practice.

Molecular phylogeny and taxonomy of colorless, filamentous sulfur bacteria of the genus Thiothrix

Comprehensive investigation combining molecular genetic techniques and comparative studies of morphological and physiological properties made it possible to resolve the disputed issue of the taxonomic status of the groups “T. nivea” and “Eikelboom type 021N” of the genus Thiothrix. The phylogenetic trees constructed on the basis of 16S rRNA and gyrB gene sequences demonstrated that members of the genus Thiothrix formed a cluster within the order Thiotrichales. According to the “ribosomal” tree, the cluster of the genus Thiothrix was divided into two main groups, I and II, corresponding to the groups “T. nivea” and “Eikelboom type 021N”. The levels of similarity between the 16S rRNA gene sequences of Thiothrix species reached 88.9–100%. On the contrary, in the “gyrase” tree, these species were not divided into “T. nivea” and “Eikelboom type 021N” groups. The levels of similarity between the amino acid sequences of the gyrB gene fragments of Thiothrix species varied from 74.5 to 99.2%. Importantly, members of the groups “T. nivea” and “Eikelboom type 021N” formed very similar 16S rRNA secondary structures in the variable region V3, where a 30-nucleotide deletion characteristic of all Thiothrix species was detected. Phenotypic analysis of the studied bacteria revealed some morphological and physiological properties shared by the groups “T. nivea” and “Eikelboom type 021N”. The data obtained indicate that members of the groups “T. nivea” and “Eikelboom type 021N” are phenotypically and genetically heterogeneous species within the single monophyletic genus Thiothrix..

Stable limited filamentous bulking through keeping the competition between floc-formers and filaments in balance

Limited filamentous bulking (LFB) was proposed to save aeration energy consumption and enhance the capacity of filaments to degrade substrates with low concentrations in activated sludge systems. Operational parameters favorable for maintaining the LFB state were investigated in an anoxic–oxic reactor treating domestic wastewater. The experiments showed that the LFB state would deteriorate with sharply decreasing temperature, reducing substrate gradients or removing anoxic zones. The balance between filaments and floc-formers could be achieved by controlling dissolved oxygen and sludge loading rates to be in optimal ranges. Eikelboom Type 0041 and CandidatusMicrothrix parvicella were the filamentous bacteria responsible for the LFB state. However, the excess growth of Eikelboom Type 021N and Sphaerotilus natans were observed when serious bulking occurred under low substrate gradients. It was demonstrated that stable maintenance of LFB for energy saving was feasible by process control and optimization.

Growth of filamentous bacteria in an enhanced biological phosphorus removal system

Activated sludge bulking caused by filamentous bacteria Eikelboom type 021N and Thiothrix spp. is reported for the first time in a multistage pilot-scale wastewater treatment plant for enhanced biological phosphorus removal (EBPR). Both types of microorganisms grew gradually reaching their maximum cumulative length within a period of 2 months. For identification of the strains, in addition to physiological observations under the microscope, Gram staining, Neisser staining and sulfur storage test took place. Results confirmed that biological nutrient removal systems may also suffer from sludge bulking. Plant configuration, wastewater composition, removal efficiency and filaments’ growth stages were examined in relation to the occurrence of the certain strains in an EBPR system.

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.

Relationships among filamentous microorganisms in rotating biological contactors

The existing relationships were studied among the different types of filamentous microorganisms that appear in the biofilm of a biological contactor system. Using the hierarchical cluster analysis it was observed that, in all the stages, Beggiatoa sp. and the Eikelboom's types 0803 and 1863 always appeared associated, while Sphaerotilus natans was always associated with the morphological type 021N. The remaining microorganisms were associated in variable forms in the plants. In addition, different association models were obtained according to the season of the year and the stage-season interaction. It has also been observed that a significant correlation exists among the filamentous microorganisms we have studied and the different physical-chemical parameters.