Plasmid-mediated bioaugmentation of sequencing batch reactors for enhancement of 2,4-dichlorophenoxyacetic acid removal in wastewater using plasmid pJP4.

Plasmid-mediated bioaugmentation was demonstrated using sequencing batch reactors (SBRs) for enhancing 2,4-dichlorophenoxyacetic acid (2,4-D) removal by introducing Cupriavidus necator JMP134 and Escherichia coli HB101 harboring 2,4-D-degrading plasmid pJP4. C. necator JMP134(pJP4) can mineralize and grow on 2,4-D, while E. coli HB101(pJP4) cannot assimilate 2,4-D because it lacks the chromosomal genes to degrade the intermediates. The SBR with C. necator JMP134(pJP4) showed 100 % removal against 200 mg/l of 2,4-D just after its introduction, after which 2,4-D removal dropped to 0 % on day 7 with the decline in viability of the introduced strain. The SBR with E. coli HB101(pJP4) showed low 2,4-D removal, i.e., below 10 %, until day 7. Transconjugant strains of Pseudomonas and Achromobacter isolated on day 7 could not grow on 2,4-D. Both SBRs started removing 2,4-D at 100 % after day 16 with the appearance of 2,4-D-degrading transconjugants belonging to Achromobacter, Burkholderia, Cupriavidus, and Pandoraea. After the influent 2,4-D concentration was increased to 500 mg/l on day 65, the SBR with E. coli HB101(pJP4) maintained stable 2,4-D removal of more than 95 %. Although the SBR with C. necator JMP134(pJP4) showed a temporal depression of 2,4-D removal of 65 % on day 76, almost 100 % removal was achieved thereafter. During this period, transconjugants isolated from both SBRs were mainly Achromobacter with high 2,4-D-degrading capability. In conclusion, plasmid-mediated bioaugmentation can enhance the degradation capability of activated sludge regardless of the survival of introduced strains and their 2,4-D degradation capacity.

Effect of fine bubbles for washing of monolith type porous ceramic membranes treating oil-in-water emulsions.

Produced water generated in the recovery of crude oil contains oil and high concentrations of salts, organic matter, and suspended solids and must therefore be treated appropriately prior to disposal. Monolithic ceramic membranes have high oil removal rates and have the advantage of being compact, having a long life, and withstanding chemicals, heat, and high cleaning pressures. Membrane fouling, however, is a significant drawback to membrane filtration. Scrubbing using air bubbles generated by a diffuser is generally used to physically clean membranes. However, monolithic ceramic membranes cannot be scrubbed using air bubbles because their fluid channels are only a few millimeters wide. Membrane washing efficiency was therefore evaluated using fine bubbles smaller than the diameter of the channels. In dead-end filtration, flushing the membrane surface with air-microbubble water or air-ultra-fine bubble (UFB) water after backwashing and air-blowing (conventional cleaning) of the channels was more efficient than conventional cleaning. Flushing with UFB water was not influenced by changes in pH that changed the zeta potential of the UFB. Membrane fouling was suppressed in crossflow filtration by mixing UFB water with feed water. There was no significant change in the diameter of the oil droplets in the feed water before and after UFB mixing. The ZP of the oil droplets peaked at around -20 mV before UFB mixing. However, the peak shifted to around -25 to -29 mV after UFB mixing.

A quantitative analysis method to determine the amount of cellulose fibre in waste sludge.

A novel quantitative analysis method for cellulose fibre was developed to understand its behaviour in biological wastewater treatment and waste sludge processes. The method developed in this study was designed using Pseudomonas aeruginosa to remove it by dissolving all the organic components except cellulose from the sludge due to needing the solubilisation of bacteria occupied almost of sludge matrix and quantifying the amount of remaining cellulose. The results of this study indicated that a combined treatment process that employed 2,000 U/L protease, 2 M hydrogen peroxide, and 2 mM potassium hydroxide after pre-treatment for floc dispersion with an ultrasonic treatment at 26 W for 1 min resulted in a solubilisation of 96% of P. aeruginosa without losing the cellulose fibre. When it was applied to the cellulose fibre added in the sludge from a municipal wastewater treatment facility, 99.5% of the cellulose fibre was recovered by using the high-speed centrifuge.

Reduction of ozone dosage by using ozone in ultrafine bubbles to reduce sludge volume.

Sludge ozonation, which promotes sludge disintegration and solubilization, is a promising technology for reducing waste sludge volume from biological wastewater treatment process. However, if this technology is to be widely adopted, reducing the energy consumption associated with ozone generation will be necessary. We used ultra-fine bubbles (UFBs) as ozone carriers to determine if their use could improve the efficiency of ozone treatment and reduce the ozone dose required. We used a spiral, liquid-type UFB generator, which can introduce UFBs directly into a sludge suspension. The death ratio of bacteria in sludge was used as an indicator of sludge reduction. The ozone requirement was reduced by UFBs. The ozone consumption required to achieve a death ratio of 80% was 15 mg-O3/g-MLSS in the sludge treated with ozone supplied by UFBs versus 25 and 45 mg-O3/g-MLSS in sludges treated with ozone supplied as a spiral, liquid-type microbubbles and by a diffuser, respectively. When mixing water ozonated with UFBs with sludge, the depth of the dead cell layer from the surface to the interior of the sludge floc was larger than that of ozonated water lacking UFBs at the same rate of ozone consumption. Ozone in UFBs kills bacteria inside the flocs. However, the fragmentation of sludge flocs by shear forces in the UFB generator made a larger contribution to the acceleration of bacterial death in sludge treated with ozone supplied by UFBs.

Changes in bacterial community structure in a full-scale membrane bioreactor for municipal wastewater treatment.

This study investigated changes in the structure and metabolic capabilities of the bacterial community in a full-scale membrane bioreactor (MBR) treating municipal wastewater. Microbial monitoring was also conducted for a parallel-running conventional activated sludge (CAS) process treating the same influent. The mixed-liquor suspended solid concentration in the MBR reached a steady-state on day 73 after the start-up. Then the MBR maintained higher rates of removal of organic compounds and nitrogen than the CAS process did. Terminal restriction fragment length polymorphism analysis revealed that the bacterial community structure in the MBR was similar to that in the CAS process at the start-up, but it became very different from that in the CAS process in the steady state. The bacterial community structure of the MBR continued to change dynamically even after 20 months of the steady-state operation, while that of the CAS process was maintained in a stable condition. By contrast, Biolog assay revealed that the carbon source utilization potential of the MBR resembled that of the CAS process as a whole, although it declined transiently. Overall, the results indicate that the bacterial community of the MBR has flexibility in terms of its phylogenetic structure and metabolic activity to maintain the high wastewater treatment capability.

Bacterial community dynamics in a full-scale municipal wastewater treatment plant employing conventional activated sludge process.

To elucidate the bacterial community dynamics in a full-scale wastewater treatment plant (WWTP) and the relatedness among bacterial communities in the influent, effluent and sludge, the structure and metabolic ability of the bacterial community throughout a full-scale WWTP employing a conventional activated sludge process was investigated during a period of 10 months. The bacterial community structure was analyzed by terminal-restriction fragment length polymorphism targeting eubacterial 16S rRNA genes, while a Biolog assay was applied to assess the metabolic ability of the activated sludge. Influent bacterial community structure was generally stable. In contrast, the bacterial community structure in the effluent was similar to that in the influent in some cases, while in other cases it was unique and differed greatly from that in the influent and sludge. These results suggest that temporal variations of the effluent bacterial community may be useful to predict the wastewater treatment performance and settleability of activated sludge. The bacterial community structure in the sludge was relatively stable and was rarely impacted by the influent populations. Biolog assay also revealed that activated sludge maintained a remarkably similar metabolic potential of organic compounds over time due to functional redundancy, in which the minor populations played a significant role.