Quorum sensing: an emerging link between temperature and membrane biofouling in membrane bioreactors.

Lab-scale membrane bioreactors (MBRs) were investigated at 12, 18, and 25 °C to identify the correlation between quorum sensing (QS) and biofouling at different temperatures. The lower the reactor temperature, the more severe the membrane biofouling measured in terms of the transmembrane pressure (TMP) during filtration. More extracellular polymeric substances (EPSs) that cause biofouling were produced at 18 °C than at 25 °C, particularly polysaccharides, closely associated with QS via the production of N-acyl homoserine lactone (AHL). However, at 12 °C, AHL production decreased, but the release of EPSs due to deflocculation increased the soluble EPS concentration. To confirm the temperature effect related to QS, bacteria producing AHL were isolated from MBR sludge and identified as Aeromonas sp., Leclercia sp., and Enterobacter sp. through a 16S rDNA sequencing analysis. Batch assays at 18 and 25 °C showed that there was a positive correlation between QS through AHL and biofilm formation in that temperature range.

The attachment potential and N-acyl-homoserine lactone-based quorum sensing in aerobic granular sludge and algal-bacterial granular sludge.

Bacteria and algae often coexist in the aerobic granular sludge (AGS) system in a photo-bioreactor, forming algal-bacterial granular sludge. In this study, the physicochemical characteristics and microbial attachment potential of the AGS and algal-bacterial granular sludge were comparatively analyzed. Results clearly showed that the larger and denser algal-bacterial granular sludge had stronger attachment potential compared to the AGS (as the control). A bioassay with Agrobacterium tumefaciens KYC55 indicated that N-acyl-homoserine lactones (AHLs) existed in both sludge types, but further investigations revealed that the relative AHL content of the algal-bacterial granular sludge obviously increased and slightly decreased during phases II and III, respectively, but was consistently higher than the AGS. Based on the EPS measurements and 3D-excitation-emission matrix (3D-EEM) fluorescence spectra analysis, the enhancement of AHL-based QS favored the hydrophobic protein production of algal-bacterial granular sludge, contributing to a good development of the granular sludge. In addition, it was also found that inhibition of AHLs resulted in the reduction of the protein content and attachment potential in algal-bacterial granular sludge, which was unfavorable to the structural stability of the granules. High-throughput sequencing analysis showed that the microbial community of AGS was different from the algal-bacterial granular sludge; specifically, algal-bacterial granulation facilitated the abundance of AHLs and EPS producers, such as the genera Acinetobacter, Chryseobacterium, and Flavobacterium.

Characterization of N-Acyl Homoserine Lactones in Vibrio tasmaniensis LGP32 by a Biosensor-Based UHPLC-HRMS/MS Method.

Since the discovery of quorum sensing (QS) in the 1970s, many studies have demonstrated that Vibrio species coordinate activities such as biofilm formation, virulence, pathogenesis, and bioluminescence, through a large group of molecules called N-acyl homoserine lactones (AHLs). However, despite the extensive knowledge on the involved molecules and the biological processes controlled by QS in a few selected Vibrio strains, less is known about the overall diversity of AHLs produced by a broader range of environmental strains. To investigate the prevalence of QS capability of Vibrio environmental strains we analyzed 87 Vibrio spp. strains from the Banyuls Bacterial Culture Collection (WDCM911) for their ability to produce AHLs. This screening was based on three biosensors, which cover a large spectrum of AHLs, and revealed that only 9% of the screened isolates produced AHLs in the defined experimental conditions. Among these AHL-producing strains, Vibrio tasmaniensis LGP32 is a well-known pathogen of bivalves. We further analyzed the diversity of AHLs produced by this strain using a sensitive bioguided UHPLC-HRMS/MS approach (Ultra-High-Performance Liquid Chromatography followed by High-Resolution tandem Mass Spectrometry) and we identified C10-HSL, OH-C12-HSL, oxo-C12-HSL and C14:1-HSL as QS molecules. This is the first report that documents the production of AHL by Vibrio tasmaniensis LGP32.

Beneficial effects of bacteria-plant communication based on quorum sensing molecules of the N-acyl homoserine lactone group.

Bacterial quorum sensing (QS) mechanisms play a crucial role in the proper performance and ecological fitness of bacterial populations. Many key physiological processes are regulated in a QS-dependent manner by auto-inducers, like the N-acyl homoserine lactones (AHLs) in numerous Gram-negative bacteria. In addition, also the interaction between bacteria and eukaryotic hosts can be regulated by AHLs. Those mechanisms gained much attention, because of the positive effects of different AHL molecules on plants. This positive impact ranges from growth promotion to induced resistance and is quite contrasting to the rather negative effects observed in the interactions between bacterial AHL molecules and animals. Only very recently, we began to understand the molecular mechanisms underpinning plant responses to AHL molecules. In this review, we gathered the latest information in this research field. The first part gives an overview of the bacterial aspects of quorum sensing. Later we focus on the impact of AHLs on plant growth and AHL-priming, as one of the most understood phenomena in respect to the inter-kingdom interactions based on AHL-quorum sensing molecules. Finally, we discuss the potential benefits of the understanding of bacteria-plant interaction for the future agricultural applications.

Decoding Microcystis aeruginosa quorum sensing through AHL-mediated transcriptomic molecular regulation mechanisms.

Acyl-homoserine lactone (AHL) serves as a key signaling molecule for quorum sensing (QS) in bacteria. QS-related genes and physiological processes in Microcystis aeruginosa remain elusive. In this study, we elucidated the regulatory role of AHL-mediated QS in M. aeruginosa. Using AHL activity extract and transcriptomic analysis, we revealed significant effects of the AHL on growth and photosynthesis. AHL significantly increased chlorophyll a (Chl-a) content and accelerated photosynthetic rate thereby promoting growth. Transcriptome analysis revealed that AHL stimulated the up-regulation of photosynthesis-related genes (apcABF, petE, psaBFK, psbUV, etc.) as well as nitrogen metabolism and ribosomal metabolism. In addition, AHL-regulated pathways are associated with lipopolysaccharide and phenazine synthesis. Our findings deepen the understanding of the QS system in M. aeruginosa and are important for gaining insights into the role of QS in Microcystis bloom formation. It also provides new insights into the prevalence of M. aeruginosa in water blooms.

Quorum sensing regulation in Microcystis aeruginosa: Insights into AHL-mediated physiological processes and MC-LR production.

Quorum sensing (QS) is a widespread regulatory mechanism in Gram-negative bacteria, primarily involving the secretion of N-acyl homoserine lactone (AHL) to facilitate population density sensing. However, the existence of QS in blue-green algae, a subset of photoautotrophic Gram-negative bacteria forming high-density communities in water blooms, remains elusive. This study delves into the unexplored realm of QS in Microcystis aeruginosa (M. aeruginosa) by investigating AHL-related regulatory mechanisms and their impact on various physiological processes. Utilizing high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and biosensors, a hitherto unknown long-chain AHL exhibiting a mass-to-charge ratio of 318 was identified in sterile M. aeruginosa cultures. Our investigation focused on discerning correlations between AHL activity fluctuations and key parameters such as microcystin (MC-LR) production, algal density, photosynthesis, buoyancy, and aggregation. Furthermore, the AHL extract was introduced during the logarithmic stage of M. aeruginosa cultures to observe the response in physiological processes. The results revealed that AHL, functioning as an autoinducer (AI), positively influenced algal growth and photosynthesis, as evidenced by the upregulated photosynthetic conversion efficiency of PSI and chlorophyll synthesis gene (psbA). AI also played a crucial role in altering surface characteristics through the synthesis of polysaccharides and proteins in EPS, subsequently promoting cell aggregation. Concomitantly, AI upregulated mcyD, enhancing the synthesis of MC-LR. Notably, our investigation pinpointed the initiation of QS in Microcystis at a density of approximately 1.22 × 10^7 cells/mL. This groundbreaking evidence underscores the regulatory role of AI in governing the physiological processes of growth, aggregation, buoyancy, and MC-LR production by activating pertinent gene expressions. This study significantly expands the understanding of QS in AHL, providing crucial insights into the regulatory networks operating in blue-green algae.

- Invited Review - Chemical signalling within the rumen microbiome.

Ruminants possess a specialized four-compartment forestomach, consisting of the reticulum, rumen, omasum, and abomasum. The rumen, the primary fermentative chamber, harbours a dynamic ecosystem comprising bacteria, protozoa, fungi, archaea, and bacteriophages. These microorganisms engage in diverse ecological interactions within the rumen microbiome, primarily benefiting the host animal by deriving energy from plant material breakdown. These interactions encompass symbiosis, such as mutualism and commensalism, as well as parasitism, predation, and competition. These ecological interactions are dependent on many factors, including the production of diverse molecules, such as those involved in quorum sensing (QS). QS is a density-dependent signalling mechanism involving the release of autoinducer (AIs) compounds, when cell density increases AIs bind to receptors causing the altered expression of certain genes. These AIs are classified as mainly being N-acyl-homoserine lactones (AHL; commonly used by Gram-negative bacteria) or autoinducer-2 based systems (AI-2; used by Gram-positive and Gram-negative bacteria); although other less common AI systems exist. Most of our understanding of QS at a gene-level comes from pure culture in vitro studies using bacterial pathogens, with much being unknown on a commensal bacterial and ecosystem level, especially in the context of the rumen microbiome. A small number of studies have explored QS in the rumen using 'omic' technologies, revealing a prevalence of AI-2 QS systems among rumen bacteria. Nevertheless, the implications of these signalling systems on gene regulation, rumen ecology, and ruminant characteristics are largely uncharted territory. Metatranscriptome data tracking the colonization of perennial ryegrass by rumen microbes suggest that these chemicals may influence transitions in bacterial diversity during colonization. The likelihood of undiscovered chemicals within the rumen microbial arsenal is high, with the identified chemicals representing only the tip of the iceberg. A comprehensive grasp of rumen microbial chemical signalling is crucial for addressing the challenges of food security and climate targets.

A sustainable strategy for effective regulation of aerobic granulation: Augmentation of the signaling molecule content by cultivating AHL-producing strains.

The positive roles of N-acyl homoserine lactone (AHL)-mediated quorum sensing (QS) in aerobic granular sludge (AGS) have been widely acknowledged. However, it is not feasible to manipulate granulation via direct addition of AHL chemicals or AHL-producing strains. Here, several strains with high AHL-producing capacity were successfully isolated from AGS. These QS strains were cultivated, mixed as a consortium, and then divided into two groups: AHLs supernatant and bacterial cells encapsulated in sodium alginate (CEBs). The potential of QS regulation, via doses of AHLs supernatant and CEBs, in accelerating granulation was evaluated. Results clearly indicated that short-term (days 21-70) addition of AHLs supernatant led to a rapid specific growth rate (0.08 d-1), compact structure without filamentous bacteria overgrowth, excellent settlement performance (SVI10 37.2 mL/g), and a high integrity coefficient (4.4%) of the granules. Sustainable release of AHLs (mainly C6- and C8-HSL) was induced by exogenous AHLs, possibly attributed to the enrichment of the genera Aeromonas and Pseudomonas. Further, tryptophan and aromatic protein substances were produced to maintain structural stability, suggesting that short-term QS regulation had long-term positive effects on the characteristics of AGS. By comparison, the addition of CEBs posed negligible or negative impact on the granulation, as evidenced by the rupture of smaller aggregates and poor characteristics of AGS. Overall, augmentation of the signaling content via addition of AHLs supernatant from QS strains is an economical and feasible regulation strategy to accelerate granulation and sustain long-term structural stability.

[Identification and Influence of Quorum Sensing on Anaerobic Ammonium Oxidation Process].

Anaerobic ammonium oxidation (ANAMMOX), an innovative nitrogen removal technology, has good prospects for applications. However, ANAMMOX bacteria grow slowly and are hardly accumulated in bioreactors. In this study, a UASB reactor inoculated with sludge from landfill leachate treatment plant was used for the start-up of ANAMMOX process. Besides, exogenous quorum sensing signals (DSF and AHL) were added to improve the adhesion of ANAMMOX sludge. The results showed that the UASB successfully started the ANAMMOX process within 150 days of operation. The total nitrogen removal rate reached 80% and the proportion of ANAMMOX bacteria rose to 20%. There was a low concentration of AHLs signal molecules in the ANAMMOX sludge. If the ß-position substituent group of AHL added was a carbonyl group (including 3-oxo-C6-HSL, 3-oxo-C8-HSL, 3-oxo-C10-HSL and 3-oxo-C12-HSL), the adhesion growth ability of the ANAMMOX sludge could be improved. In the case of dosing with AHL molecules without ß-position substituent groups, only C6-HSL and C12-HSL could promote the adhesion of ANAMMOX sludge. The additions of C8-HSL, C10-HSL and DSF all had negative effects on the adhesion of ANAMMOX sludge.

Complete genome sequencing of Pandoraea pnomenusa RB38 and Molecular Characterization of Its N-acyl homoserine lactone synthase gene ppnI.

In this study, we sequenced the genome of Pandoraea pnomenusa RB38 using Pacific Biosciences RSII (PacBio) Single Molecule Real Time (SMRT) sequencing technology. A pair of cognate luxI/R homologs was identified where the luxI homolog, ppnI, was found adjacent to a luxR homolog, ppnR1. An additional orphan luxR homolog, ppnR2, was also discovered. Multiple sequence alignment and phylogenetic analysis revealed that ppnI is an N-acyl homoserine lactone (AHL) synthase gene that is distinct from those of the nearest phylogenetic neighbor viz. Burkholderia spp. High resolution tandem mass spectrometry (LC-MS/MS) analysis showed that Escherichia coli BL21 harboring ppnI produced a similar AHL profile (N-octanoylhomoserine lactone, C8-HSL) as P. pnomenusa RB38, the wild-type donor strain, confirming that PpnI directed the synthesis of AHL in P. pnomenusa RB38. To our knowledge, this is the first documentation of the luxI/R homologs of the genus Pandoraea.

Cell adhesion, ammonia removal and granulation of autotrophic nitrifying sludge facilitated by N-acyl-homoserine lactones.

In this study, six N-acyl-homoserine lactone (AHL) molecules (C6-HSL, C8-HSL, C10-HSL, 3-oxo-C6-HSL, 3-oxo-C8-HSL and 3-oxo-C10-HSL) were each dosed into a bioreactor and seeded using autotrophic nitrifying sludge (ANS). The effects of the AHLs on cell adhesion, nitrification and sludge granulation were investigated. The results indicated that the efficiencies of cell adhesion and ammonia removal both had a close correlation with the side chain length and ß position substituent group of the AHLs. The best-performing AHL in terms of accelerating bacterial attached-growth was 3-oxo-C6-HSL, whereas C6-HSL outperformed the others in terms of the ammonia degradation rate. The addition of 3-oxo-C6-HSL or C6-HSL increased the biomass growth rate, microbial activity, extracellular proteins and nitrifying bacteria, which can accelerate the formation of nitrifying granules. Consequently, selecting AHL molecules that could improve bacteria in attached-growth mode and nitrification efficiency simultaneously will most likely facilitate the rapid granulation of nitrifying sludge.

Diversity and N-acyl-homoserine lactone production by Gammaproteobacteria associated with Avicennia marina rhizosphere of South Indian mangroves.

The diversity of N-acyl-homoserine lactone (AHL)-producing rhizosphere bacterial community associated with Avicennia marina in the mangrove ecosystems of South India was investigated. Approximately 800 rhizobacteria were isolated from A. marina, and they were screened for the production of AHL using two biosensors, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens NTL4 (pZLR4). Among the total isolates screened, 7% of the rhizobacteria showed positive induction for AHL signals. The BOX-PCR profile of 56 positive isolates represented 11 distinct genotypic groups. Phylogenetic analyses of the 16S rRNA sequences of 16 representatives showed that the isolates belonged to the class Gammaproteobacteria, which represented six different genera: Pseudomonas, Aeromonas, Vibrio, Photobacterium, Serratia and Halomonas. The study also identified three AHL-producing species, namely, Photobacterium halotolerans MSSRF QS48, Vibrio xiamenensis MSSRF QS47 and Pseudomonas sp. MSSRF QS1 that had not been reported previously. AHL profiling by TLC detected short chains C4, C6 and C8-HSL, and long chains C10 and C12-HSL with both unsubstituted and substituted side chains among the 16 representative AHL positives. This is the first report concerning the diversity of AHL-producing Gammaproteobacteria from mangrove ecosystems exhibiting diverse AHL profiles.