Characterization and Significance of Sub-Visible Particles and Colloids in a Submerged Anaerobic Membrane Bioreactor (SAnMBR).

The distribution, composition and morphological structure of subvisible particles and colloids (0.01-10 µm) in the supernatant of a lab-scale submerged anaerobic membrane bioreactor (SAnMBR), and their role in membrane fouling, was investigated. Photometric analysis showed that the supernatant and membrane foulants were dominated by particles and colloids (0.45-10 µm), which accounted for over 90% of the total organics (proteins and polysaccharides). Excitation-emission matrix (EEM) fluorescence spectra and monosaccharide analysis showed that these particles and colloids were rich in fluorescent proteins, rhamnose, ribose and arabinose, all of which could be related to cellular and extracellular substances. Fluorescence and scanning electron microscopy confirmed the presence of bacterial cells in/on the subvisible particles and colloids. The microparticles (5-10 µm) were primarily composed of Streptobacilli and/or filamentous bacteria in the form of microcolonies, while the submicrometer particles and colloids (1-5 µm and 100 kDa-1 µm) had more free/single cocci and bacilli. The ratio of live/dead cells varied in different size-fractions, and the particles (1-10 µm) contained more live cells compared with the colloids (100 kDa-1 µm). Our findings suggest that bacterial cells in/on the particles and colloids could have an important effect on fouling in SAnMBRs as they represent pioneering species attaching to membranes to form fouling layers/biofilm. Such insights reveal that previous foulant-characterization studies in MBRs tended to overestimate organic fouling, while the biofouling induced by these bacteria in/on the particles and colloids was overlooked.

Biofouling control potential of quorum quenching anaerobes in lab-scale anaerobic membrane bioreactors: Foulants profile and microbial dynamics.

Indigenously isolated anaerobes encoding four quorum quenching (QQ) enzymes were applied in immobilized- and bioaugmented forms for their implications on membrane foulants, microbial taxa, and biofouling control. Two identical anaerobic membrane bioreactors (AnMBRs) with different immobilizing media, i.e. silica-alginate (AnMBR-Si) and hollow fiber-alginate (AnMBR-Hf), were sequentially operated for two conventional and three QQ based phases. The synergistic addition of QQ anaerobes in free cells and the immobilized form prolonged the membrane filtration operation by 172 ± 29% and 284 ± 12% in AnMBR-Si and AnMBR-Hf, respectively. Biocake with low surface coverage was prominent during QQ application compared to conventional phases. Despite the better control of AHLs (3OC6-, C6-, 3OC8, C8, and C10-HSL) and AI-2 at various points of QQ phases, the QQ consortium could not maintain a low concentration of signals for longer period. Therefrom, quenching of targeted signal molecules instigate the dominance of microbial species bearing non-targeted quorum sensing mechanism. The QQ significantly altered the biofilm-forming community in mixed liquor, while the members with robust signal transduction systems became dominant to counteract the QQ mechanism and were the ultimate cause of biofouling. The improved methane content in biogas and increased methanogens composition during QQ phases demonstrated the synergism of exogenous and immobilized QQ as the most viable option for long-term AnMBR operation.

The selective pressure of quorum quenching on microbial communities in membrane bioreactors.

In conventional membrane bioreactor (MBR) treatment systems, Gram-negative bacterial population appears to be always outnumbered Gram-positive community. Thereby, acyl homoserine lactones (AHLs), major signaling molecules utilized by Gram-negative bacteria, have been targeted for biofouling control in quorum quenching (QQ) based studies. This study investigated the impact of AHL and autoinducer-2 (AI-2)-degrading QQ consortium on the selective accumulation of microbial communities in a QQ MBR (MBR-QQb). The results show that addition of the QQ consortium (in the form of beads) increased the filtration time of MBR-QQb by 3.5 times. The distribution of mixed liquor extracellular polymeric substances (EPS), especially the tightly bound (TB) proteinous EPS and the floc size were strongly affected by the QQ activity, and the endless 'battle' between QQ and quorum sensing (QS). More importantly, QQ induced the significant suppression of Gram-negative bacterial community. The average abundance of Gram-positive bacteria at the genus level in the biocake of MBR-QQb (51%) was significantly higher than that of the control MBR (11%) and the MBR with vacant beads (28%). These findings suggest that an unintended condition is created to favor the growth of Gram-positive bacteria in QQ MBRs, resulting in a distinct microbial social network in both bulk sludge and biocake.

Size-dependent microbial diversity of sub-visible particles in a submerged anaerobic membrane bioreactor (SAnMBR): Implications for membrane fouling.

Sub-visible particles, an often-overlooked fine particle (0.45-10 µm) with a size between sludge solids and soluble microbial products (SMP), have recently been identified as a critical foulant in anaerobic membrane bioreactors (AnMBRs), and our recent new insights into the size-fractionation and composition of sub-visible particles in AnMBRs have enabled fouling to be understood in more depth. Here, we investigated the microbial diversity of the sub-visible particles in three size fractions (i.e., 5-10, 1-5, and 0.45-1 µm) from bulk and cake solutions in a lab-scale AnMBR, and their fouling potential was further explored based on their filtration behavior and biofilm formation. Results show that with decreasing particle size, a significant shift in microbial communities was observed for the sub-visible particles in both bulk and cake solutions; (a) with notable decreases in filamentous microbes in the order SJA-15, GCA004, and Anaerolineales of phylum Chloroflexi, and, (b) with substantial increases in sulfate-reducing bacteria (i.e., the family Syntrophobacteraceae, genus DCE29 of family Thermodesulfovibrionaceae, Desulfovibrio, and Geobacter). More importantly, the filamentous microbes associated with micro-particles (5-10 µm) led to higher cake fouling resistances while free living cells in the form of colloidal particles (0.45-1 µm) induced severer pore blocking. Moreover, the micro-particles had an enhanced capacity to favor biofilm formation (OD595 = 1.0-2.5, categorized as highly positive), thus potentially aggravating biofouling. This work advances our knowledge on the effect of particle size on communities and underlying fouling behavior of microbes associated with fine particles in AnMBRs.

Insights into quorum quenching mechanisms to control membrane biofouling under changing organic loading rates.

A quorum quenching (QQ) consortium comprised of both acyl homoserine lactones (AHLs)- and autoinducer-2 (AI-2)-degrading bacteria, either immobilized in polymer-coated alginate beads or in liquid suspension, was examined for fouling control in lab-scale MBRs under both steady and changing organic loading rates (OLRs). Under steady conditions the QQ consortium retarded biofouling by a factor of 3. However, a continuous increase in OLR vastly reduced the effectiveness of QQ bacteria; the biofouling was retarded only by factors of 1.4-1.8. A significant increase in extracellular polymeric substance (EPS), especially loosely-bound EPS in mixed liquor together with an increase in polysaccharide content up to 4 times in EPS resulted from the increase in OLR, was attributed to the impaired QQ efficacy. In control MBRs, cake layer resistance was the major factor (>60%) contributing to the increased trans-membrane pressure, as compared with pore blockage resistance and intrinsic membrane resistance. In contrast, the pore blockage resistance became dominant in QQ MBRs (>40%).