Microthrix Parvicella Research Articles

Control of Filamentous Bacteria overgrowth by Changing the Hydraulic Flow Distribution of a Step-feed Biological Nitrogen Removal (SFBNR) Process-based WWTP: A Full-scale Study

To facilitate the wastewater recycling process while adhering to environmental and industrial regulations, it becomes imperative to meet stringent standards. This necessitates the effective elimination of organic substances, with particular emphasis on nitrogen-based compounds. To this end, activated sludge methods, particularly the step-feed biological nitrogen removal (SFBNR) process, are employed. Although various operational measures are employed in SFBNR systems, including the elimination of complex substrates, modifications in aeration conditions, the utilization of piston hydraulic flow, and the appropriate distribution of substrates, these systems remain susceptible to the excessive growth of filamentous bacteria. The current study aimed to examine the change in the distribution of the hydraulic flow entering this system on a real scale over a period of five years. It was observed that the quality of sludge settling improved and accordingly the overgrowth of filamentous Microthrix parvicella declined, thereby enhancing the quality of the effluent.

Microbial assessment of foaming dynamics in full-scale WWTP anaerobic digesters

Research on foaming processes in wastewater treatment plants (WWTPs) anaerobic digesters is increasing since it is a common phenomenon that can seriously affect digesters performance and treatment cost. This paper presents data from a full-scale experience of foam monitoring and antifoam reagents dosing as well as microbial community structure. The association of foaming episodes with microbial population structure and operational parameters has improved understanding of foaming episodes and their causes. During this long-term study (273 days), two foaming episodes occurred in the anaerobic digesters under study. Microbial community analysis showed that the first foaming episode is correlated with the proliferation of ST-12K33 (midas_s_22), which could be related to variations in the hydraulic retention time, a decrease in inflow solids concentration, and a decrease in digesters total alkalinity. The second foaming episode coincide with a clear migration of Ca. Microthrix parvicella from the activated sludge (bulking episodes) to the anaerobic digesters and its capacity to metabolize substrate under anaerobic conditions. Laboratory experiments showed that an increase in anaerobic digestion temperature and sludge pre-treatments have a positive impact on foam biodegradability and, therefore, they could be used as foam mitigation strategies. Finally, a techno-economic analysis showed that foaming episodes had an over cost of about 250 € d−1 and dropped the WWTP energy self-sufficiency from 67 to 63 %.

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.

Taxonomic Structure Evolution, Chemical Composition and Anaerobic Digestibility of Microalgae-Bacterial Granular Sludge (M-BGS) Grown during Treatment of Digestate

The liquid fraction from the dewatering of digested sewage sludge (LF-DSS) represents a major processing complication for wastewater treatment facilities, thus necessitating new and effective methods of LF-DSS neutralization. This pilot-scale study examined the evolution of a Chlorella sp. monoculture into microalgal-bacterial granular sludge (M-BGS) during treatment of LF-DSS in a hybrid photo-bioreactor (H-PBR). The M-BGS reached a stable taxonomic and morphological structure after 60 days of H-PBR operation. The biomass was primarily composed of Chlorella sp., Microthrix parvicella, and type 1851 and 1701 filamentous bacteria. A greater abundance of bacteria led to a faster-growing M-BGS biomass (to a level of 4800 ± 503 mgTS/dm3), as well as improved TOC and COD removal from the LF-DSS (88.2 ± 7.2% and 84.1 ± 5.1%). The efficiency of N/P removal was comparable, since regardless of the composition and concentration of biomass, it ranged from 68.9 ± 3.1% to 71.3 ± 3.1% for N and from 54.2 ± 4.1% to 56, 2 ± 4.6% for P. As the M-BGS taxonomic structure evolved and the C/N ratio improved, so did the anaerobic digestion (AD) performance. Biogas yield from the M-BGS peaked at 531 ± 38 cm3/gVS (methane fraction = 66.2 ± 2.7%). It was found that final effects of AD were also strongly correlated with the N and TOC content in the substrate and pH value. A mature M-BGS significantly improved settleability and separability through filtration.

Operation of a pilot-scale lipid accumulation technology employing parameters to select Microthrix parvicella for biodiesel production from wastewater

Wastewater treatment plants (WWTPs) may play a crucial role in shifting to a zero-emission future by becoming more sustainable and contributing to the circular economy (CE). Recovered lipids from urban sewage can serve as a raw material for biofuel production contributing to a waste reduction, mitigation of natural resources depletion and reinforcing security and energy independence. A novel, pilot-scale lipid accumulation technology (LAT) employing parameters to select M. parvicella for the biofuel/biodiesel production was implemented on a side stream of an urban WWTP. The LAT proved its concept as the average amount of extracted lipids accumulated in the bioreactors was three-fold higher when compared to the lipids existing in activated sludge. The average transesterification of extracted lipids to biodiesel resulted in a 1.6 % yield, meaning that from 1 kg of dried sludge, 16 g of biodiesel could be formed. The biodiesel produced complies with European standard specifications (EN14214).

Mobilome-driven segregation of the resistome in biological wastewater treatment.

Biological wastewater treatment plants (BWWTP) are considered to be hotspots for the evolution and subsequent spread of antimicrobial resistance (AMR). Mobile genetic elements (MGEs) promote the mobilization and dissemination of antimicrobial resistance genes (ARGs) and are thereby critical mediators of AMR within the BWWTP microbial community. At present, it is unclear whether specific AMR categories are differentially disseminated via bacteriophages (phages) or plasmids. To understand the segregation of AMR in relation to MGEs, we analyzed meta-omic (metagenomic, metatranscriptomic and metaproteomic) data systematically collected over 1.5 years from a BWWTP. Our results showed a core group of 15 AMR categories which were found across all timepoints. Some of these AMR categories were disseminated exclusively (bacitracin) or primarily (aminoglycoside, MLS and sulfonamide) via plasmids or phages (fosfomycin and peptide), whereas others were disseminated equally by both. Combined and timepoint-specific analyses of gene, transcript and protein abundances further demonstrated that aminoglycoside, bacitracin and sulfonamide resistance genes were expressed more by plasmids, in contrast to fosfomycin and peptide AMR expression by phages, thereby validating our genomic findings. In the analyzed communities, the dominant taxon Candidatus Microthrix parvicella was a major contributor to several AMR categories whereby its plasmids primarily mediated aminoglycoside resistance. Importantly, we also found AMR associated with ESKAPEE pathogens within the BWWTP, and here MGEs also contributed differentially to the dissemination of the corresponding ARGs. Collectively our findings pave the way toward understanding the segmentation of AMR within MGEs, thereby shedding new light on resistome populations and their mediators, essential elements that are of immediate relevance to human health.

Microbiological aspects of the aeration tanks of an activated sludge treatment plant in dysfunction: Consequences on its treatment performance

The present study was conducted in a sewage treatment plant, called "El Kouwaer", located in the department of Mascara, North-West of Algeria. Mainly, the objective of this investigation is to evaluate the relevant settling and pollution parameters and to assess the proliferation level of filamentous microbial communities during the dysfunction period to further the effectiveness of the operative treatment. Sludge parameter values were estimated such us Settling Test, Diluted Sludge Volume Index (DSVI), and Total Suspended Solids (TSS) content to set up the sludge classification, the performance analysis of treatment was conducted by computing pollution indicators and removal efficiencies of organic pollution, as well as the Principal Component Analysis (PCA) of dysfunction parameters. The results showed that in aeration tanks 65% of samples represented a maximum Filament Index (FI) of 6. Diluted Sludge Volume Index (DSVI) was greater than 300 mg/L in 23% of samples. A high level of filamentous bacteria biodiversity was also noted during the assessment of the (FI) including Microthrix parvicella, Bogiotae, and Sphaerotilus natans. The effluent in the biological unit was characterized by BOD5/COD ratio of 0.17 and COD removal efficiency of 53.32%. Those results demonstrated that the sewage treatment plant is frequently affected by bulking problems. This study had confirmed the importance of continuous monitoring of different process parameters like Wastewater Biodegradability Index, Dissolved Oxygen, filamentous flora, and Dilute Sludge Volume Index (DSVI) to avoid irreversible damage at the level of a wastewater treatment plant.

Seasonal microbial community dynamics complicates the evaluation of filamentous bulking mitigation strategies in full-scale WRRFs

The activated sludge wastewater treatment process has been thoroughly researched in more than 100 years, yet there are still operational challenges that have not been fully resolved. Such a challenge is the control of filamentous bulking caused by the overgrowth of certain filamentous bacteria. In this study, we tested different mitigation strategies to reduce filamentous bulking, caused by two common filamentous genera found in full-scale water resource recovery facilities (WRRF), Candidatus Microthrix and Candidatus Amarolinea. PAX dosing, ozone addition, hydrocyclone implementation, and the addition of nano-coagulants were tested as mitigation strategies in four parallel treatment lines in a full-scale WRRF over three consecutive years. Unexpectedly, the activated sludge settleability was not affected by any of the mitigation strategies. Some of the strategies appeared to have a strong mitigating effect on the two filamentous species. However, detailed analyses of the microbial communities revealed strong recurrent seasonal variations in all four lines, including the control line which masked the real effect. After removing the effect of the seasonal variation by using a time-series decomposition approach, it was clear that the filamentous bacteria were mostly unaffected by the mitigation strategies. Only PAX dosing had some effect on Ca. Microthrix, but only on one species, Ca. Microthrix subdominans, and not on the most common Ca. Microthrix parvicella. Overall, our study shows the importance of long-term monitoring of microbial communities at species level to understand the normal seasonal pattern to effectively plan and execute full-scale experiments. Moreover, the results highlight the importance of using parallel reference treatment lines when evaluating the effect of mitigation strategies in full-scale treatment plants.

Effects of silica fume powder modified by oleic acid on the settleability of bulking sludge

ABSTRACT Modified silica fume powder with oleic acid through coupling agent was prepared based on the in situ utilizing long-chain fatty acids (LCFA) properties of Microthrix parvicella (M. parvicella) in the activated sludge system. The modification was confirmed by XRD and infrared spectrum. The contact angle analysis showed that the modification gave the silica fume powder a hydrophobic surface. The modified silica fume powder had a good combination with M. parvicella from the SEM and Gram staining measurements. The addition of modified silica powder has a certain effect on the settling capacity of sludge, but has little effect on the sludge treatment capacity, while the SVI dropped from 400.1 to 100.0 mL/g. These suggested that the modified silica fume powder could be used as an excellent weight-increasing agent to inhibit sludge bulking.

Biophysical Strategies as an Effective Means to Eliminate Microthrix Parvicella from Activated Sludge System

Biophysical Strategies as an Effective Means to Eliminate Microthrix Parvicella from Activated Sludge System

Pilot Scale Operation Applying Optimum Growth Conditions for Microthrix Parvicella to Enhance Lipid Accumulation from Wastewater for Biodiesel Production

Pilot Scale Operation Applying Optimum Growth Conditions for Microthrix Parvicella to Enhance Lipid Accumulation from Wastewater for Biodiesel Production

Structural and functional changes in bacterial communities during sludge foaming in the sub-plateau MSTPs

Sludge foaming has been a persistent ailment in worldwide municipal sewage treatment plants (MSTPs). In particular, this problem frequently occurs and lasts throughout the winter and spring season in the sub-plateau MSTPs. Aims to explore the causes of sludge foaming under sub-plateau circumstance, the correlation between the structural and functional changes in bacterial communities and sewage properties was conducted using Illumina high-throughput sequencing technology. The results from 16S rRNA gene sequencing shown that the relative abundances of Actinobacteria (39.5 % ± 4.8 %) and Firmicutes (10.5 % ± 1.0 %) in the foaming sludge were remarkably higher than that in the non-foaming sludge. The abundances of Candidatus Microthrix parvicella (29.8 % ± 6.9 %), Trichococcus (6.5 % ± 1.0 %), and Ornithinibacter (3.5 % ± 0.6 %) were increased after sludge foaming, and which are considered as the dominated foaming-causative bacterial assemblages by the linear discriminant analysis (LDA). Spearman’s correlation analysis and redundancy analysis (RDA) also agreed that lower temperature and dissolved oxygen (DO) levels, higher lipid and protein contents were significantly correlated with sludge foaming. In addition, the predictive functional profiling indicated that the enzymes and their coding genes of the bacterial communities are mainly involved in the fast proliferation and lipid metabolism, and which abundance are highly upregulated during foaming period. These results suggested that the sludge foaming was strongly linked to the Actinobacteria and Firmicutes communities and higher lipid and protein content in the sewage under sub-plateau circumstance. The findings might be important for developing operating strategies to maintain the desired microbial community pattern and improve the performance of MSTPs.

Simulation and experimental study of fluorescence labeled polyphosphate in microthrix parvicella

To study the binding mechanism of 4′,6-diamidino-2-phenylindole (DAPI) and polyphosphate (Poly-P) and find fluorescent dyes which can dye Poly-P better, the interaction model of DAPI and Poly-P was calculated by the self-consistent-charge, density functional tight-binding (SCC-DFTB-D) method, and the binding sites of DAPI and Poly-P were analyzed. Further, Cy3, Rhodamine 6G and Fluorescein, which are structurally similar to DAPI, were selected to analyze their interactions with Poly-P. The binding energies and frontier orbital properties of the complexes were analyzed. These four fluorescent dyes were further used to dye the activated sludge smear and observe the fluorescence property. The Simulation results show that the N-containing indole ring structures in DAPI plays an important role in the interaction with Poly-P; the binding energies for DAPI, Cy3, Rhodamine 6G and Fluorescein with Poly-P are −42.6, −165.4, −34.7 and −28.9 kcal/mol, respectively. The frontier orbital properties for the complexes were studied, which further indicates that the interactions between Cy3, Rhodamine 6G and Poly-P are stronger than that of Fluorescein and Poly-P. The experimental results showed that Cy3 had excellent dyeing effect on Poly-P and could recognize them, while Fluorescein could not dye Poly-P. The experimental results were in good agreement with those predicted by simulation, which verified the correctness of our calculation method and provided a new strategy for finding more reliable, more sensitive and more economical fluorescent dyes capable of dyeing Poly-P.

Roles of bacteriophages, plasmids and CRISPR immunity in microbial community dynamics revealed using time-series integrated meta-omics

Viruses and plasmids (invasive mobile genetic elements (iMGEs)) have important roles in shaping microbial communities, but their dynamic interactions with CRISPR-based immunity remain unresolved. We analysed generation-resolved iMGE–host dynamics spanning one and a half years in a microbial consortium from a biological wastewater treatment plant using integrated meta-omics. We identified 31 bacterial metagenome-assembled genomes encoding complete CRISPR–Cas systems and their corresponding iMGEs. CRISPR-targeted plasmids outnumbered their bacteriophage counterparts by at least fivefold, highlighting the importance of CRISPR-mediated defence against plasmids. Linear modelling of our time-series data revealed that the variation in plasmid abundance over time explained more of the observed community dynamics than phages. Community-scale CRISPR-based plasmid–host and phage–host interaction networks revealed an increase in CRISPR-mediated interactions coinciding with a decrease in the dominant ‘Candidatus Microthrix parvicella’ population. Protospacers were enriched in sequences targeting genes involved in the transmission of iMGEs. Understanding the factors shaping the fitness of specific populations is necessary to devise control strategies for undesirable species and to predict or explain community-wide phenotypes.

Correlating sludge constituents with digester foaming risk using sludge foam potential and rheology.

Foam potential and viscometer ramp tests (VRTs) were conducted for three municipal wastewater treatment plants to determine if these methods can relate to mechanisms of foaming to physical and biological constituents in sludge. At all plants, digester volatile solids (VS) concentration correlated (R2 > 0.41) with increases in plastic viscosity, a VRT parameter corresponding to foaming risk. Plastic viscosity also correlated with foam-causing bacteria Gordonia (R2 = 0.38). Foam potential test values increased with Microthrix parvicella (R2> 0.28). For one plant, suspected foam-causing bacteria Mycobacterium negatively correlated with parameters representing foam risk. Microscopic filament counting correlated (R2 = 0.97) with quantitative polymerase chain reaction (qPCR) for Gordonia, suggesting that the more accessible counting method can reliably quantify foam-causing bacteria. Foam potential tests and VRTs resulted in plant-specific correlations with foam-related constituents. Therefore, these tests may provide useful evidence when investigating causes of digester foam events.

Factors influencing Candidatus Microthrix parvicella growth and specific filamentous bulking control: A review.

Candidatus Microthrix parvicella has been frequently detected as the dominant filamentous bacteria in bulking sludge and thus seriously affects the stable operation of activated sludge processes. The extremely low growth rate of Ca. M. parvicella and its sensitivity to environmental variations greatly limit the development of effective techniques to control filamentous bulking. Based on previous investigations, a variety of restrictive substrates, operating and culture conditions, environmental factors and other potential inhibitors have varying degrees of impact on the growth of this microorganism. This review systematically summarizes the key factors affecting Ca. M. parvicella growth with a focus on the influencing mechanism. Recent filamentous bulking control strategies are also critically reviewed and discussed. Additionally, research needs for the next few years are proposed with the aim of establishing effective and specific control strategies for filamentous sludge bulking.

Insights into the effects of carbon source on sequencing batch reactors: Performance, quorum sensing and microbial community.

Effects of carbon source on the performance, quorum sensing (QS) and microbial communities in the sequencing batch reactors were investigated in this work. Among the chosen carbon source, sodium acetate (R1), glucose (R2), starch (R3) and Tween 80 (R4), sodium acetate was the best carbon source for nutrient removal, while starch was favorable for inducing the sludge bulking, and Tween 80 was beneficial to the production of extracellular polymeric substances (EPS) and proliferation of Microthrix parvicella. Additionally, the R2 value of linear correlation between sludge settleability and particle size in four reactors followed an order of R1 > R2 > R3 > R4. Moreover, Person correlation analysis showed that various significant correlations were observed in reactors fed with different carbon sources and the QS mainly mediated the production and component of EPS. High-throughput sequencing analysis revealed that the carbon source affected microbial communities and the Canonical correspondence analysis results indicated that QS related to microbial communities. It was inferred that the interactions between microbial communities and QS affected system performance.

Factors affecting SVI in small scale WWTPs

This paper analyses factors associated with bulking in 195 small scale wastewater treatment plants (WWTPs) in Estonia. Operational data from each plant were collected and analysed statistically. The key factors associated with bulking were infiltration into sewage pipes, the type and purpose of process reactor, operational practices and influent characteristics. Both anaerobic fraction and volumetric fraction of the anaerobic reactor compared to the aerobic reactor resulted in a positive correlation with sludge volume index (SVI) <150 ml/g values. Good operation and maintenance practice as well as an operator's competence play a crucial role in bulking prevention. Using the 30 minute settling test (V30) as the single process control parameter can mislead an operator's judgement in process control strategies and cause effluent violations. Misjudgements in process control decisions can lead to unwanted conditions in small WWTPs (e.g. excessive chemical addition favoured bulking). Use of instrumentation, control and automation helped to keep the process conditions more stable and reduce the probability of bulking. Analyses of variance showed that the factors associated with Microthrix parvicella growth were long solids retention time (SRT), low food-to-microorganism ratio (F/M) and lack of carbon content compared against nitrogen and phosphorus contents.

Monitoring, isolation and characterization of Microthrix parvicella strains from a Chinese wastewater treatment plant.

Microthrix parvicella is a filamentous bacterium that frequently causes severe bulking events in wastewater treatment plants (WWTPs) worldwide. In this study, sludge properties and dynamics of filamentous bacteria in a Beijing WWTP seasonally suffering from M. parvicella bulking were continuously monitored over a duration of 15 months, and the correlations between M. parvicella and operating parameters were evaluated. The predominance of M. parvicella was observed at low temperatures (14-18.8 °C) with the relative abundance of around 30% (estimated by both qPCR and FISH analysis). Using micromanipulation technology, 545 filaments of M. parvicella were micromanipulated from bulking sludge (SVI > 180 mL g-1) on six different media. After 3-month purification and enrichment, six strains, phylogenetically closely related to Candidatus Microthrix parvicella, were successfully acquired on R2A medium (20 °C) in pure cultures. Considering the limitation and extremely slow growth rate of M. parvicella filaments, newly isolated strains represent valuable sources for further investigations on the physiology and behavior of this filamentous bacterium, with the focus on the establishment of bulking control strategy.

Decision tree for identification and prediction of filamentous bulking at full-scale activated sludge wastewater treatment plant

This study attempted to model sludge bulking in a full-scale wastewater treatment plant operated as a 3-stage Phoredox process. Principal component analysis and a regression tree model were employed to describe the correlations between influent wastewater characteristics and operational conditions (as inputs) and sludge volume index (SVI) as an output. A classification tree model was used to determine the environmental factors that affected the proliferation of filamentous microorganisms. Fluorescent in situ hybridisation analysis identified filamentous species of Microthrix parvicella, Thiothrix I & II, and Eikelboom Types 0041, 0092, and 021 N. It was found that SVI increased with an increment in sludge retention time, but it negatively correlated with soluble chemical oxygen demand (sCOD) and ammonium-nitrogen. The dominance of Microthrix parvicella was observed with a decline in temperature below 15.5 °C, causing an increase in SVI during the winter and spring seasons. The overgrowth of Thiothrix could be linked to the unbalanced ratio between readily biodegradable COD and nutrient species. The filament Type 0092 contributed to high SVI, and it prevailed with a decrease in food-to-microorganisms ratio below 0.08 1/d. Based on the satisfactory training, validation, and generalization procedures, the proposed models could be applied for the prediction of sludge bulking episodes.