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This study discovered a strong correlation between the autoinducer-2 (AI-2)-mediated quorum sensing (QS) with the performance of a submerged anaerobic membrane bioreactor during its recovery from a pentachlorophenol (PCP) shock: a decrease in AI-2 levels coincided with a reduction in volatile fatty acid concentrations, and corresponded significantly to a decrease in the relative abundance of Firmicutes, and to an increase in the relative abundance of Bacteroidetes and Synergistetes. Further batch experiments with the addition of an AI-2-regulating Escherichia coli mutant culture showed that a reduction in AI-2 levels resulted in the highest biogas production rate during a PCP shock. In contrast, an increase in AI-2 levels via addition of the E. coli wild type strain or an AI-2 precursor showed no obvious effects on biogas production. These results suggest that the AI-2 level in anaerobic sludge was governed primarily by Firmicutes, and the AI-2-mediated QS partially regulates the toxic shock response of anaerobic sludge via tuning the activities of Firmicutes and Synergistetes. A decrease in the AI-2 level might reduce acetogenesis and favor hydrogenotrophic methanogenesis, thus resulting in less VFA accumulation and higher methane production during the PCP shock. This study is the first of this type that exploits the role of quorum sensing in the toxic shock response of anaerobic sludge; it demonstrates a novel approach to shortening the recovery period of anaerobic processes via manipulating the AI-2-mediated QS. National Research Foundation (NRF) The project was supported and administrated by the Environment & Water Industry Programme Office (EWI) for and on behalf of the National Research Foundation Board (NRF), Singapore (EWI-IRIS-0807). The project was also supported by the Shantou University Scientific Research Foundation for Talents (No.: NTF16015) and the National Natural Science Foundation of China (No.: 51750110514).

Biological wastewater treatment processes (WWTPs), by nature of their reliance on biological entities to degrade organics and sometimes remove nutrients, are vulnerable to toxicants present in their influent. Various toxicity measurement methods have been adopted for biological WWTPs, but most are performed off-line, and cannot be adapted to on-line monitoring tools to provide an early warning for WWTP operators. However, the past decade has seen a rapid expansion in the research and development of biosensors that can be used for toxicity assessment of aquatic environments. Some of these biosensors have also been shown to be effective for use in biological WWTPs. Nevertheless, more research is needed to: examine the sensitivity of assays and sensors based on single organisms to various toxicants and develop a matrix of biosensors or a biosensor incorporating multiple organisms that can protect WWTPs; test the micro fuel cell (MFC)-based biosensors with real wastewaters and correlate the results with the well-established oxygen uptake rate (OUR)-based or CH4-based toxicity assay; and, develop advanced data processing methods for interpreting the results of on-line toxicity sensors in real WWTPs to reduce the noise due to the normal fluctuation in influent quality and quantity.

Pseudomonas-derived regulators DmpR and XylR are structurally and mechanistically related σ54-dependent activators that control transcription of genes involved in catabolism of aromatic compounds. The binding of distinct sets of aromatic effectors to these regulatory proteins results in release of a repressive interdomain interaction and consequently allows the activators to promote transcription from their cognate target promoters. The DmpR-controlled Po promoter region and the XylR-controlled Pu promoter region are also similar, although homology is limited to three discrete DNA signatures for binding σ54RNA polymerase, the integration host factor, and the regulator. These common properties allow cross-regulation of Pu and Po by DmpR and XylR in response to appropriate aromatic effectors. In vivo, transcription of both the DmpR/Po and XylR/Pu regulatory circuits is subject to dominant global regulation, which results in repression of transcription during growth in rich media. Here, we comparatively assess the contribution of (p)ppGpp, the FtsH protease, and a component of an alternative phosphoenolpyruvate-sugar phosphotransferase system, which have been independently implicated in mediating this level of regulation. Further, by exploiting the cross-regulatory abilities of these two circuits, we identify the target component(s) that are intercepted in each case. The results show that (i) contrary to previous speculation, FtsH is not universally required for transcription of σ54-dependent systems; (ii) the two factors found to impact the XylR/Pu regulatory circuit do not intercept the DmpR/Po circuit; and (iii) (p)ppGpp impacts the DmpR/Po system to a greater extent than the XylR/Pu system in both the nativePseudomonas putidaand a heterologousEscherichia colihost. The data demonstrate that, despite the similarities of the specific regulatory circuits, the host global regulatory network latches onto and dominates over these specific circuits by exploiting their different properties. The mechanistic implications of how each of the host factors exerts its action are discussed.

This work aimed to investigate the promotive effect of Ag modification on C-TiO2 for visible light photocatalytic disinfection of microorganisms. Ag/C-TiO2 was prepared by a facile one-step synthesis approach through thermal oxidation of AgNO3-impregnated TiC precursor. The self-doped carbon exists as both interstitial and substitutional carbon to collectively enhance the visible light photocatalytic absorption. The loaded silver nanoparticles were uniformly distributed on the TiO2 surface in the Ag0 metallic state, further enhancing the visible light absorption due to surface plasmon resonance yet inhibiting the charge carrier recombination by conduction band electron trapping. The band structure and a tentative visible light photocatalytic disinfection mechanism of Ag/C-TiO2 with four pathways were thus proposed, and the enhanced visible light photocatalytic performance was demonstrated for disinfection of Escherichia coli (E. coli) and Enterococcus faecalis (E. faecalis) in comparison with C-TiO2. This improvement was more pronounced with increasing Ag loading in the range of 0.5–5.0 wt%, which can be attributed to the improved visible light absorption and good charge carrier separation due to Ag modification as well as to the inherent bactericidal effect of metallic Ag. ASTAR (Agency for Sci., Tech. and Research, S’pore)

When studying interspecies interactions in a bacterial consortium, it may be desirable to analyze one species' transcriptional response as influenced by the other species. We developed a dual fluorescence system of Escherichia coli for Fluorescence-Activated Cell Sorter (FACS)-based analysis for such a purpose. First, we generated E. coli SCC1 strain, which constitutively expresses green fluorescent protein (GFP), but otherwise showed no observable difference from the parent strain MG1655 with respect to morphology, growth, and FACS-analyzed side- and forward-scatter profiles. Next, to analyze transcriptional response, plasmids carrying promoters of interest fused to a red fluorescent protein (AsRed2) reporter, were introduced into strain SCC1. Quantification of promoter activities of araB, lacZ, fadB and rpoE via AsRed2 reporter verified that the induction levels are similar between MG1655 and SCC1 strains. In mixtures and co-cultures, GFP expression of E. coli SCC1 allowed it to be separated from non-E. coli species by FACS to purity levels of 96.7-100.0%. When a mixture of E. coli SCC1 carrying promoter-AsRed2 fusion and a non-E. coli strain was analyzed by FACS, it enabled (i) distinction of E. coli SCC1 from the non-E. coli strain, (ii) analysis of the E. coli promoter activity via AsRed2 expression and (iii) identification of transcriptional heterogeneity within the E. coli population. Co-cultures of E. coli SCC1 with Klebsiella pneumoniae and/or Enterococcus faecalis analyzed by FACS showed that E. coli fadB and rpoE transcription were differentially influenced by partner species.

Transcription from Pseudomonas-derived -24, -12 Po promoter of the pVI150-encoded dmp operon is mediated by the sigma 54-dependent DmpR activator in response to the presence of aromatic pathway substrates in the medium. However, global regulatory mechanisms are superimposed on this regulatory system so that the specific response to aromatic effectors is absent in cultures until the stationary phase is reached. Here we genetically dissect the system to show that the growth phase response is faithfully mimicked by a minimal system composed of the dmpR regulatory gene and the Po promoter regulatory region and can be reproduced in heterologous Escherichia coli. Using this system, we show that the growth phase-dependent DmpR-mediated response to aromatic compounds is limited to fast-growing cultures. Thus, during exponential growth of cultures in minimal media containing different carbon sources, the response to aromatics is immediate, while the response is suppressed in cultures grown on rich media until the exponential-to-stationary phase transition. Elements known to be involved in the DmpR-mediated transcription from Po were analyzed for the ability to influence the growth phase response. Most dramatically, overexpression of DmpR was shown to completely abolish the growth phase response, suggesting that a negatively acting factor may mediate this level of regulation. The possible mechanism of action and integration (of the specific regulation of the dmp operon-encoded catabolic enzymes with the physiological status of the bacteria are discussed.