Bioaugmented Sequencing Batch Reactor Research Articles

Enhanced treatment of synthetic wastewater by bioaugmentation with a constructed consortium

Bioaugmentation by adding well-functioning mixed microorganism consortia represents a potentially useful approach to improve contaminant removal in wastewater treatment plants (WWTPs). However, unfavorable environmental conditions (i.e., low temperatures) can severely inhibit microbial activity, drawing our attention to constructing cold-tolerant microorganism preparations and investigating their availability in practical applications. Here we screened four in situ functional isolates from the activated sludge of secondary sedimentation tanks in WWTPs to construct a psychrophilic microbial consortium, which was used to perform bioaugmentation for enhanced removal of nitrogen and phosphorus under low temperatures. The consortium was established by cocultivation of four isolates, characterized by 16 S rRNA as the COD-degrading bacterium Aeromonas sp. Z3, aerobic denitrifying bacterium Acinetobacter sp. HF9, nitrifying bacterium Klebsiella sp. X8, and polyphosphate-accumulating bacterium Pseudomonas sp. PC5 respectively. The microorganism preparation was composed of Z3, HF9, X8, and PC5 under the ratio of 1: 1: 3: 1, which can exert optimal pollutant removal under the conditions of 12 °C, 6.0–9.0 pH, 120–200 r‧min−1, and a dosage of 5% (V/V). A 30-day continuous operation of the bioaugmented and control sequencing batch reactors (SBRs) was investigated, and the bioaugmented SBR showed a shorter start-up stage and a more stable operating situation. Compared to the control SBR, the COD, NH4+-N, TN, and TP removal efficiency of the bioaugmented SBR increased by an average of 7.95%, 9.05%, 9.54%, and 7.45% respectively. The analysis of the microbial community revealed that the introduced isolates were dominant in the activated sludge and that functional taxa such as Proteobacteria, Bacteroidota, and Actinobacteria were further enriched after a period of bioaugmentation. The study provides some basis and guidance for the practical application of how to strengthen the stable operation of WWTPs under low temperatures.

Changes in Wastewater Treatment Performance and the Microbial Community during the Bioaugmentation of a Denitrifying Pseudomonas Strain in the Low Carbon–Nitrogen Ratio Sequencing Batch Reactor

The low carbon–nitrogen ratio (C/N) of influent wastewater results in the insufficient carbon source for the process of denitrification in urban wastewater treatment plants (WWTPs). A denitrifying bacterial strain, Pseudomonas sp. JMSTP, was isolated and demonstrated effective denitrification ability under a low C/N ratio of 1.5–4 (w/w) in anoxic conditions. Sequencing batch reactor (SBR) studies were conducted to test the bioaugmentation of JMSTP on total nitrogen (TN) removal under the influent COD/N ratio of 3/1. After the second bioaugmentation, the TN of effluent in the bioaugmented SBR was significantly lower than that in the control SBR. Redundancy analysis results showed that there was a positive correlation between Pseudomonas sp. abundance and TN removal in the bioaugmented SBR. Microbial community analysis showed that, especially after the second bioaugmentation, the abundance of Pseudomonas sp. decreased rapidly, but it was still much higher than that in the control SBR. Correlation network analysis showed that after the addition, Pseudomonas sp. had no significant co-occurrence relationship with other native bacteria, owing to the quick increase and decrease. Our results suggest that JMSTP shows the potential to enhance TN removal through bioaugmentation. Since the effect of bioaugmentation gradually diminishes, further research is still needed to investigate its long-lasting applications.

Microbial community and function evaluation in the start-up period of bioaugmented SBR fed with aniline wastewater

An enhanced sequencing batch reactor (SBR) system was developed to treat synthetic wastewater rich in 600 mg/L aniline. The aniline degradation efficiency was almost 100%, and the total nitrogen (TN) removal rate was more than 50%. Metagenomics technology revealed the community structure, functional genes and metabolic mechanism during the start-up of the enhanced reactor. Sequencing results showed that Proteobacteria, Bacteroidetes, Chloroflexi and Actinobacteria were dominant phylum. The proportion of degradation of aromatic compounds function increased gradually, but the proportion of nitrogen metabolism function changed little. Functional genes involved in aniline degradation including benA-xylX and dmpB/xylE were detected. The functional genes of nitrogen metabolism were involved in complete nitrification, traditional denitrification, assimilation nitrate reduction and dissimilation nitrate reduction. The functional contribution analysis and network analysis showed that the cooperation and competition of Thauera, Delftia, Diaphorobacter, Micavibrio and Azoarcus ensured the effective removal of aniline and nitrogen.

Improvement in nitrogen removal and changes in community structure in a sequencing batch reactor bioaugmented with P. stutzeri strain XL-2

The aim of this work was to study the bioaugmentation of P. stutzeri strain XL-2 in activated sludge to improve nitrogn removal from wastewater with the guide of growth kinetics. When 4250 mg/L COD and 80 mg/L NH4+-N were applied, the TN removal efficiency in a bioaugmented sequencing batch reactor (SBRXL) achieved 95%, while that in the control reactor (SBRC) without strain XL-2 was only 84% (P < 0.05). The microbial community analysis demonstrated that strain XL-2 was successfully bioaugmented in SBRXL, and increasing influent COD concentration promoted its abundance. Influent COD concentration played a dominant role in affecting community structure, while the bioaugmentation of strain XL-2 had much less impact on the community structure. Combined with principal coordinates analysis, redundancy analysis and FAPROTAX, the improvement of TN removal was mainly achieved by the bioaugmentation of strain XL-2, which played a major role in promoting aerobic denitrification.

Effects of dissolved oxygen concentrations on a bioaugmented sequencing batch rector treating aniline-laden wastewater: Reactor performance, microbial dynamics and functional genes

This work compared the efficiencies and internal reasons for aniline removal in a bioaugmented sequencing batch reactor at elevated dissolved oxygen (DO) concentrations. Nearly complete aniline removal was achieved while medium DO of 2.65 mg/L was optimal for subsequent nitrogen removal via heterotrophic nitrification-aerobic denitrification. Apart from the highest bacterial diversity richness, favorable DO condition largely enriched putatively aniline degrader, nitrifiers and denitrifiers. Further evidence from qPCR confirmed that moderate DO significantly stimulated the functional genes including aniline degrading gene tad, nitrifying genes amoA, hao, and denitrifying genes nirS and napA, respectively. This study indicates that the significant enrichment of key microorganisms and effective functional genes under optimal DO is the inner mechanism for reliable aniline degradation and subsequent nitrogen removal in the activated sludge reactor.

Application of Bioaugmentation Strategy in Biodegradation of Di-n-octyl Phthalate (DOP) in Sequencing Batch Reactor (SBR)

In order to enhance the degradation of Di-n-octyl phthalate (DOP).One bacterial strain was inoculated in SBR. Bioaugmentation by inoculating DOP-degrading bacteria effectively shortened the start-up of SBRs and significantly enhanced DOP degradation in bioreactors (p<0.05), 92.3% DOP removal rate was achieved in consortium bioaugmented SBR, which was double of control SBR. The DOP removal in control reactor was only 37%. The results from this study suggest bioaugmentation is an effective and feasible approach for DOP waste water treatment in practical engineering.

Genetic Bioaugmentation of Activated Sludge with Dioxin-Catabolic Plasmids Harbored by Rhodococcus sp. Strain p52.

Horizontal transfer of catabolic plasmids is used in genetic bioaugmentation for environmental pollutant remediation. In this study, we examined the effectiveness of genetic bioaugmentation with dioxin-catabolic plasmids harbored by Rhodococcus sp. strain p52 in laboratory-scale sequencing batch reactors (SBRs). During 100 days of operation, bioaugmentation decreased the dibenzofuran content (120 mg L-1) in the synthetic wastewater by 32.6%-100% of that in the nonbioaugmented SBR. Additionally, dibenzofuran was removed to undetectable levels in the bioaugmented SBR, in contrast, 46.8 ± 4.1% of that in the influent remained in the nonbioaugmented SBR after 96 days. Moreover, transconjugants harboring pDF01 and pDF02 were isolated from the bioaugmented SBR after 2 days, and their abilities to degrade dibenzofuran were confirmed. After 80 days, the copy numbers of strain p52 decreased by 3 orders of magnitude and accounted for 0.05 ± 0.01% of the total bacteria, while transconjugants were present at around 106 copies mL-1 sludge and accounted for 8.2 ± 0.3% of the total bacteria. Evaluation of the bacterial community profile of sludge by high-throughput 16S rRNA gene sequencing revealed that genetic bioaugmentation led to a bacterial community with an even distribution of genera in the SBR. This study demonstrates the promise of genetic bioaugmentation with catabolic plasmids for dioxins remediation.

Identification and characterization of four bacterial strains isolated from a bioaugmented sequencing batch reactor

Identification and characterization of four bacterial strains isolated from a bioaugmented sequencing batch reactor

Bioaugmentation for treating transient or continuous p-nitrophenol shock loads in an aerobic sequencing batch reactor

Bioaugmentation with an enriched microbial population was applied in an aerobic sequencing batch reactor (SBR) receiving transient or continuous shock loads of p-nitrophenol (PNP). The effect of the amount of biomass added for bioaugmentation was assessed by using two different dosages (2% or 5% w/w of the total biomass in the seeded SBR). In both cases, total PNP removal was achieved during the transient PNP shock load occurring after bioaugmentation. However, after a long PNP starvation period the only experiment still showing total PNP removal during a second PNP shock load was the one where a dosage of 5% w/w was applied. The results suggested that the dosage is a key factor for the implementation of a successful bioaugmentation strategy. In addition, the performance of a bioaugmented SBR receiving a continuous PNP shock load was enhanced when compared to a non-bioaugmented SBR.

Bioaugmentation with Gordonia strain JW8 in treatment of pulp and paper wastewater

The Gordonia strain JW8 was successfully isolated, characterized, and tested for bioaugmentation of pulp and paper wastewater. With significant degradation of alkaline lignin, JW8 has the potential to render pulp and paper wastewater more biodegradable, which is of much interest to wastewater treatment. The sequencing batch reactor (SBR) inoculated with JW8 significantly enhanced the organic pollutants removal of pulp and paper wastewater compared to the control, achieving the best removal rates of 96.4% and 87.8% for biochemical oxygen demand (BOD) and chemical oxygen demand (COD), respectively. A considerable decrease of BOD/COD ratio in wastewater was also achieved in the bioaugmented SBR. However, a drawback caused by JW8 inoculation was the resultant high sludge volume index and biological foaming and bulking, which can be controlled by food/mass adjustment. The denaturing gradient gel electrophoresis analysis suggested that bioaugmentation with JW8 had a slight effect on the microbial dynamics, but more research is needed to understand the relationship between the microbial dynamics and pollutant removal.

Effects of solids retention time on the performance of bioreactors bioaugmented with a 17β-estradiol-utilizing bacterium, Sphingomonas strain KC8

This study investigated the performance of lab-scale sequencing batch reactors (SBRs) that were inoculated with nitrifying activated sludge and bioaugmented with a Sphingomonas strain KC8 (a 17β-estradiol-degrading bacterium). The bioaugmented SBRs were supplied with synthetic wastewater (average initial total organic carbon (TOC) = 175 mg L −1 and average initial ammonia–N = 25 mg L −1) and daily dose of 17β-estradiol (1 mg L −1) and operated under three solid retention times (SRTs) of 5, 10, and 20 d. After three times periods of the operating SRTs, the overall removal of TOC (>87%) and ammonia (>91%) was similar in all the SBRs. Higher 17β-estradiol removals (>99%) were observed for the SBRs. Neither estrogens nor estrogenic activity was detected in the treated water, except some samples from the SBR operating under 5 d of SRT. The ratios of known estrogen degraders ( Sphingomonas strain KC8 and ammonia-oxidizing bacteria) and amoA gene to the total bacterial population decreased as SRT increased, suggesting the presence of unknown estrogen-degraders in SBRs operating at SRT = 10 and 20 d. Real-time-terminal-restriction fragment length polymorphism analysis showed that the evenness of microbial community structures was not affected by the SRT; while, the diversity indices suggest that longer SRTs might lead to more diverse microbial community structure. Overall, the results suggested that bioaugmented bioreactors operating at long SRTs (10 and 20 d) were effective in removing 17β-estradiol to the non-estrogenic treatment endpoint.

Analysis of denitrifier community in a bioaugmented sequencing batch reactor for the treatment of coking wastewater containing pyridine and quinoline

The denitrifier community and associated nitrate and nitrite reduction in the bioaugmented and general sequencing batch reactors (SBRs) during the treatment of coking wastewater containing pyridine and quinoline were investigated. The efficiency and stability of nitrate and nitrite reduction in SBR was considerably improved after inoculation with four pyridine- or quinoline-degrading bacterial strains (including three denitrifying strains). Terminal restriction fragment length polymorphism (T-RFLP) based on the nosZ gene revealed that the structures of the denitrifier communities in bioaugmented and non-bioaugmented reactors were distinct and varied during the course of the experiment. Bioaugmentation protected indigenous denitrifiers from disruptions caused by pyridine and quinoline. Clone library analysis showed that one of the added denitrifiers comprised approximately 6% of the denitrifier population in the bioaugmented sludge.

Bioaugmentation treatment for coking wastewater containing pyridine and quinoline in a sequencing batch reactor

Two pyridine-degrading bacteria and two quinoline-degrading bacteria were introduced for bioaugmentation to treat the coking wastewater. Sequencing batch reactors (SBRs) were used for a comparative study on the treatment efficiency of pyridine, quinoline, and chemical oxygen demand. Results showed that the treatment efficiency with coking-activated sludge plus a mixture of the four degrading bacteria was much better than that ones with coking-activated sludge only or mixed degrading bacteria only. Moreover, a 52-day continuous operation of the bioaugmented and general SBRs was investigated. The bioaugmented SBR showed better treatment efficiency and stronger capacity to treat high pyridine and quinoline shock loading. The general SBR failed to cope with the shock loading, and the biomass of the activated sludge decreased significantly. In order to monitor the microbial ecological variation during the long-term treatment, the bacterial community in both reactors was monitored by the amplicon length heterogeneity polymerase chain reaction technique. The diversity of the bacterial community decreased in both reactors, but the introduced highly efficient bacteria were dominant in the bioaugmented SBR. Our experiment showed clearly that the use of highly efficient bacteria in SBR process could be a feasible method to treat wastewater containing pyridine or/and quinoline.

Bioaugmentation of aerobic sludge granules with a plasmid donor strain for enhanced degradation of 2,4-dichlorophenoxyacetic acid

Aerobic sludge granules pre-grown on glucose were bioaugmented with a plasmid pJP4 carrying strain Pseudomonas putida SM1443 in a fed-batch microcosm system and a lab-scale sequencing batch reactor (SBR) to enhance their degradation capacity to 2,4-dichlorophenoxyacetic acid (2,4-D). The fed-batch test results showed that the bioaugmented aerobic granule system gained 2,4-D degradation ability faster and maintained a more stable microbial community than the control in the presence of 2,4-D. 2,4-D at the initial concentration of about 160 mg/L was nearly completely removed by the bioaugmented granule system within 62 h, while the control system only removed 26% within 66 h. In the bioaugmented SBR which had been operated for 90 days, the seeded aerobic granules pre-grown on glucose successfully turned into 2,4-D degrading granules through bioaugmentation and stepwise increase of 2,4-D concentration from 8 to 385 mg/L. The granules showed a compact structure and good settling ability with the mean diameter of about 450 μm. The degradation kinetics of 2,4-D by the aerobic granules can be described with the Haldane kinetics model with V max = 31.1 mg 2,4-D/gVSS h, K i = 597.9 mg/L and K s = 257.3 mg/L, respectively. This study shows that plasmid mediated bioaugmentation is a feasible strategy to cultivate aerobic granules degrading recalcitrant pollutants.

Microbial community dynamics in bioaugmented sequencing batch reactors for bromoamine acid removal

Sphingomonas xenophaga QYY with the ability to degrade bromoamine acid (BAA) was previously isolated from sludge samples. The enhancement of BAA removal by strain QYY in sequencing batch reactors (SBRs) was investigated in this study. The results showed that augmented SBRs exhibited stronger abilities to degrade BAA than the non-augmented control one. In order to estimate the relationship between community dynamics and function of augmented SBRs, a combined method based on fingerprints (ribosomal intergenic spacer analysis, RISA) and 16S rRNA gene sequencing was used. The results indicated that the microbial community dynamics were substantially changed, and the introduced strain QYY was persistent in the augmented systems. This study suggests that it is feasible and potentially useful to enhance BAA removal using BAA-degrading bacteria, such as S. xenophaga QYY.

Biodegradation of 2,4-dichlorophenol in sequencing batch reactors augmented with immobilized mixed culture

2,4-Dichlorophenol (2,4-DCP) degrading mixed culture was immobilized in polyvinyl alcohol jel beads and supplemented to sequencing batch reactors (SBR) to treat 2,4-DCP containing wastewater. Impacts of bioaugmentation level on the performance of bioaugmented systems were studied. Results show that inoculum size affected the start-up time of the SBR systems. For the non-augmented SBR system, nine days was needed for the system to start-up, whereas it only took six, four, three and two days for the SBRs with 1.9%, 3.7%, 5.6% and 9.3% immobilized culture, respectively. In addition, bioaugmented SBR systems demonstrated stronger capacity to cope with high 2,4-DCP shock loading than the control system. The control SBR failed to treat 2,4-DCP at 166 mg/l in influent, while the SBR with 1.9% inoculation could successfully cope with 2,4-DCP at 166 mg/l, but failed at 250 mg/l, and the SBR with 3.7%, 5.6% and 9.3% immobilized culture could successfully degrade 250 mg/l 2,4-DCP in feed. Furthermore, the contributions to the removal of 2,4-DCP by the introduced and indigenous culture in an augmented SBR system at various operation stages were investigated. It was found that augmented culture played the primary role in degrading 2,4-DCP at the beginning of system start-up, but after one-month operation, both the indigenous and the introduced culture posed strong ability to degrade 2,4-DCP.

Application of the polymerase chain reaction (PCR) and reverse transcriptase / PCR for determining the fate of phenol-degrading Pseudomonas putida ATCC 11172 in a bioaugmented sequencing batch reactor

The impact of bioaugmentation on the efficacy of an existing microbial population to detoxify phenol in a laboratory-scale sequencing batch reactor was evaluated. Phenol degradation and the persistence and expression of the catabolic dmpN gene were studied for 44 days using a combination of conventional monitoring approaches and molecular techniques. Following addition of the phenol-degrading bacterium, Pseudomonas putida ATCC 11172, which converts phenol to catechol by the aerobic meta-cleavage pathway, phenol removal␣in the bioaugmented reactor increased and was maintained at 95 %–100 %. In the unaugmented control reactor, decreased phenol removal was observed. Correspondingly, dmpN DNA, characteristic of P. putida ATCC 11172, and its expression were detected in activated sludge biomass from the bioaugmented reactor for over 41 days. The results of this study show that (i) bioaugmentation provides stability in phenol degradation, and (ii) monitoring with molecular techniques such as the polymerase chain reaction (PCR) and reverse transcriptase/PCR can successfully assess the state of a bacterium used in bioaugmentation.