QS-related Gene Research Articles

Isolation of a quorum quenching bacterium effective to various acyl-homoserine lactones: Its quorum quenching mechanism and application to a membrane bioreactor

Biofouling, caused by microbial biofilm formation on the membrane surface and in pores, is a major operational problem in membrane bioreactors (MBR). Many quorum quenching (QQ) bacteria have been isolated and applied to MBR to reduce biofouling. However, for more effective MBR biofouling control, novel approaches for isolating QQ bacteria and applying them in MBR are needed. Therefore, Listeria grayi (HEMM-2) was isolated using a mixture of different N-acyl homoserine lactones (AHLs). HEMM-2 degraded various AHLs, regardless of the length and oxo group in the carbon chain, with quorum sensing (QS) inhibition ratios of 47–61%. This QQ activity was attributed to extracellular substances in HEMM-2 cell-free supernatant (CFS). Furthermore, the HEMM-2 CFS negatively regulated QS-related gene expression, inhibiting Pseudomonas aeruginosa and activated sludge-biofilm formation by 53–75%. Surprisingly, when the HEMM-2 CFS was directly injected into a laboratory-scale MBR system, biofouling was not significantly affected. Biofouling was only controlled by cell suspension (CS) of HEMM-2, indicating the importance of QQ bacteria in MBR. The HEMM-2 CS increased operation time to reach 0.4 bar, a threshold transmembrane pressure for complete biofouling, from 315 h to 371 h. Taken together, HEMM-2, which is effective in the degradation of various AHLs, and its applicable method to MBR may be considered a potent approach for controlling biofouling and understanding the behavior of QQ bacteria in MBR systems.

Rapid start-up of partial nitrification reactor by exogenous AHLs and Vanillin combined with intermittent aeration

Quorum sensing (QS) and quorum quenching (QQ) are common phenomena in microbial systems and play an important role in the nitrification process. However, rapidly start up partial nitrification regulated by N-acyl-homoserine lactones (AHLs)-mediated QS or QQ has not been reported. Hence, we chose N-butyryl homoserine lactone (C4-HSL) and N-hexanoyl homoserine lactone (C6-HSL) as the representative AHLs, and Vanillin as the representative quorum sensing inhibitor (QSI) combined intermittent aeration to investigate their effects on the start-up process of partial nitrification. The start-up speed in the group with C4-HSL or C6-HSL addition was 1.42 or 1.26 times faster than that without addition, respectively. Meanwhile, the ammonium removal efficiency with C4-HSL or C6-HSL addition was increased by 13.87 % and 17.30 % than that of the control group, respectively. And, partial nitrification could maintain for a certain period without AHLs further addition. The increase of Nitrosomonas abundance and ammonia monooxygenase (AMO) activity, and the decrease of Nitrobacter abundance and nitrite oxidoreductase (NXR) activity were the reasons for the rapid start-up of partial nitrification in the AHLs groups. Vanillin addition reduced AMO and hydroxylamine oxidoreductase (HAO) activity, and increased Nitrobacter abundance and NXR activity, thus these were not conducive to achieving partial nitrification. Denitrifying bacteria (Hydrogenophaga, Thauera and Aquimonas) abundance increased in the Vanillin group. QS-related bacteria and gene abundance were elevated in the AHLs group, and reduced in the Vanillin group. Function prediction demonstrated that AHLs promoted the nitrogen cycle while Vanillin enhanced the carbon cycle. This exploration might provide a new technical insight into the rapid start-up of partial nitrification based on QS control.

Quorum Quenching-Mediated Biofilm Mitigation on Functionalized Ultrafiltration Membranes via Atom Transfer Radical Polymerization

Quorum sensing (QS) critically affects biofilm formation during membrane filtration, which can be manipulated for membrane biofouling mitigation. This study demonstrates a quorum quenching strategy for combating membrane biofouling by functionalizing poly(vinylidene fluoride) (PVDF) membranes via atom transfer radical polymerization of a QS inhibitor, vanillin. Poly(vanillin) brushes are successfully grafted on the membrane surface, which is verified by extensive membrane characterization. The effective biofilm inhibition ability and mitigation mechanism of poly(vanillin) brushes are evaluated by analyzing the QS-related gene expression and acyl-homoserine lactone (AHL) signal molecule concentrations. We show that poly(vanillin) brushes can inhibit the bacterial QS system and decrease the AHL concentration to mitigate membrane biofouling. In dynamic filtration experiments, the vanillin-grafted PVDF membrane exhibits a 50% increase in stable flux compared to that of the pristine PVDF membrane. Our results provide an alternate strategy for mitigating membrane biofouling by regulating the bacterial QS system via inhibitor-functionalized membranes.

Antibacterial, anti-biofilm and anti-quorum sensing activities of Artemisia dracunculus essential oil (EO): a study against Salmonella enterica serovar Typhimurium and Staphylococcus aureus.

The study evaluates the effect of Artemisia dracunculus essential oil (EO) on two pathogenic bacteria Salmonella enterica serovar Typhimurium and Staphylococcus aureus and Vero cell line. To evaluating the anti-biofilm potential of the EO, a microtiter-plate test (MtP) and scanning electron microscopy (SEM) were performed. The quorum-sensing inhibitory properties were examined by QS-related gene expression at sub-MIC concentrations of Artemisia dracunculus EO. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole) test was used to determine the cytotoxicity potential of the EO against the Vero cell line and finally, the major components of the EOs were determined using Gas chromatography-mass spectrometry (GC-MS) analysis. The minimum inhibitory concentration (MIC) of the tested EO against S. Typhimurium and S. aureus were 2.5 and 1.25μl/ml, respectively. In addition, the minimum bactericidal concentration was 5 and 2.5μl/ml for S. Typhimurium and S. aureus, respectively. Both MtP and SEM showed an acceptable inhibitory and disruption effect of the EO on the biofilm formation at Sub-MIC concentrations. Significant downregulation of luxS, pfs, and hld genes by treatment with MIC/2 concentration of A. dracunculus EO was observed. The IC50 value of A. dracunculus EO against Vero cells was 20μl/ml. The main detected compound using GC-MS was estragole (methyl chavicol or tarragon) (64.94%). Anti-biofilm, QSI activity, and non-toxicity of A. dracunculus EO reported for the first time in this study propose the use of these plant compounds as alternatives to antibiotics and chemical additives.

1-(4-Amino-2-hydroxyphenyl)ethanone from Phomopsis liquidambari showed quorum sensing inhibitory activity against Pseudomonas aeruginosa.

Phomopsis liquidambari S47 is an endophytic fungus isolated from the leaves of Punica granatum. Here, we are the first to report a quorum sensing (QS) inhibitor 1-(4-amino-2-hydroxyphenyl)ethanone (AHE) isolated and identified from the metabolites of P. liquidambari S47. Exposure to AHE at sub-MIC concentrations notably suppressed the secretion of acyl-homoserine lactones and virulence factors in Pseudomonas aeruginosa PAO1. To investigate the metabolic variations of P. aeruginosa PAO1 exposed to AHE, magnetic resonance imaging-based metabolomic analysis was performed. AHE treatment created a disturbance in the QS system by suppressing the expressions of QS-related genes. The disturbed QS system resulted in the inhibited activity of antioxidant enzymes and thus enhanced oxidative stress. The vegetable infection assay showed that the virulence of P. aeroginosa PAO1 was attenuated which could be due to the impacts to the amino acid and nucleotide metabolism by enhanced oxidative stress. These findings suggest that AHE has a potential to become an antivirulence "agent" to tackle P. aeruginosa infection. KEY POINTS: • AHE treatment inhibited AHL secretion and virulence factors production. • AHE treatment aggravated oxidative stress and disturbed metabolism. • AHE suppressed QS-related gene expressions and reduced virulence of P. aeruginosa.

Metagenomic insights into quorum sensing in membrane-aerated biofilm reactors for phenolic wastewater treatment

ABSTRACT Quorum sensing (QS) is of crucial importance for the formation and performance of biofilms adhered to aerated membranes. In this study, the QS-related genes in membrane-aerated biofilm reactors (MABR) for phenolic wastewater treatment were investigated through high-throughput metagenomic sequencing. Results showed that numerous regulatory QS-related genes were associated with the production of signals including acyl-homoserine lactones (AHLs) and diguanylate monophosphate (c-di-GMP), indicating that the biofilms were potentially regulated by the AHL-mediated QS and c-di-GMP-mediated QS systems. Species and functional contribution analysis demonstrated that Pseudomonas, Achromobacter, Rhodococcus, Granulicoccus and Thauera were the key QS-related gene carriers. Redundancy analysis and Spearman correlation analysis showed that high influent phenolic loading gave rise to a high relative abundance of QS bacteria within the biofilm community. Thus, QS-related genes likely play an important role in strengthening biofilm resistance to phenolics, as well as the removal of phenolic contaminants.

Inhibitory Effect of Two Traditional Chinese Medicine Monomers, Berberine and Matrine, on the Quorum Sensing System of Antimicrobial-Resistant Escherichia coli.

The quorum sensing (QS) system controls bacterial biofilm formation, which is highly related to the virulence and resistance of pathogens. In the present study, the effect of two traditional Chinese medicine (TCM) monomers, berberine and matrine, on biofilm formation and QS-related gene expression of antimicrobial-resistant (AMR) Escherichia coli strains was investigated by laser scanning confocal microscopy (LSCM) observation and real-time PCR. The results indicated a roughly positive relationship between biofilm formation ability and antimicrobial resistance. LSCM observation showed that berberine and matrine inhibited biofilm formation of AMR E. coli strains at 1/2 minimal inhibitory concentration (MIC) (1/2 MIC berberine at OD630: 0.1020; 1/2 MIC matrine: OD630: 0.1045); furthermore, abnormal cell morphology such as rounded and elongated cells was also observed. This finding was consistent with the downregulation of QS-related genes: luxS, pfS, sdiA, hflX, motA, and fliA. At 1/2 MIC and 1/4 MIC concentrations of berberine, a significant downregulation of luxS, pfS, hflX, ftsQ, and ftsE was observed. The results indicate that berberine and matrine can inhibit biofilm formation by inhibiting the QS system and that berberine is more effective than matrine.

Ibuprofen-mediated potential inhibition of biofilm development and quorum sensing in Pseudomonas aeruginosa

AimsPseudomonas aeruginosa is one of the leading causes of opportunistic and hospital-acquired infections worldwide, which is frequently linked with clinical treatment difficulties. Ibuprofen, a widely used non-steroidal anti-inflammatory drug, has been previously reported to exert antimicrobial activity with the specific mechanism. We hypothesized that inhibition of P. aeruginosa with ibuprofen is involved in the quorum sensing (QS) systems. Main methodsCFU was utilized to assessed the growth condition of P. aeruginosa. Crystal violent staining and acridine orange staining was used to evaluate the biofilm formation and adherence activity. The detection of QS virulence factors such as pyocyanin, elastase, protease, and rhamnolipids were applied to investigation the anti-QS activity of ibuprofen against P. aeruginosa. The production of 3-oxo-C12-HSL and C4-HSL was confirmed by liquid chromatography/mass spectrometry analysis. qRT-PCR was used to identify the QS-related gene expression. Furthermore, we explored the binding effects between ibuprofen and QS-associated proteins with molecular docking. Key findingsIbuprofen inhibits P. aeruginosa biofilm formation and adherence activity. And the inhibitory effects of ibuprofen on C4-HSL levels were concentration-dependent (p < 0.05), while it has no effect on 3-oxo-C12-HSL. Moreover, ibuprofen attenuates the production of virulence factors in P. aeruginosa (p < 0.05). In addition, the genes of QS system were decreased after the ibuprofen treatment (p < 0.05). Of note, ibuprofen was binding with LuxR, LasR, LasI, and RhlR at high binding scores. SignificanceThe antibiofilm and anti-QS activity of ibuprofen suggest that it can be a candidate drug for the treatment of clinical infections with P. aeruginosa.

Quorum sensing signals enhance the electrochemical activity and energy recovery of mixed-culture electroactive biofilms

The impacts of exogenous or endogenous quorum sensing (QS) signaling molecules on mixed-culture electroactive biofilms (EABs), especially extracellular polymeric substances (EPS) and exoelectrogens using direct electron transfer mechanism inside EABs are poorly understood. This research focuses on the influence of acylhomoserine lactones (AHLs), the most common QS signaling molecules for gram negative bacteria, on mixed-culture EABs. Results indicated that both exogenous and endogenous AHLs played the role as regulators to improve the electrochemical activities of EABs. The energy recovery of MFCs increased from 20.5% ± 3.9% to 28.3% ± 4.1% with endogenous AHLs and further rose to 36.2% ± 5.1% with exogenous AHLs, and the start-up period of MFCs shortened from 13 days to 10 days with endogenous AHLs and further reduced to 4 days in the presence of exogenous AHLs. The influences of exogenous and endogenous AHLs were non-instantaneous. They improved some intrinsic properties, i.e. the electrode-associated biomass, the biofilm compactness and the ratio of live/dead cells to obtain superior EABs. Meanwhile, both endogenous and exogenous AHLs increased the concentration and redox activities of EPS. Besides, endogenous AHLs enhanced the diversity of EPS components. Noteworthily, the relative abundance of Geoboacter sp. which is the typical microbe using direct electron transfer mechanism is raised by exogenous AHLs, though so far neither known chemical QS-related gene nor protein has been reported in this genus. These findings will increase the current understanding of QS in EABs and open up an opportunity for regulating mixed-culture MFCs via QS.

Role of the luxS Gene in Initial Biofilm Formation by Streptococcus mutans

Quorum sensing (QS) is a process by which bacteria communicate with each other by secreting chemical signals called autoinducers (AIs). Among Gram-negative and Gram-positive bacteria, AI-2 synthesized by the LuxS enzyme is widespread. The aim of this study was to evaluate the effect of QS luxS gene on initial biofilm formation by Streptococcus mutans. The bacterial cell surface properties, including cell hydrophobicity (bacterial adherence to hydrocarbons) and aggregation, which are important for initial adherence during biofilm development, were investigated. The biofilm adhesion assay was evaluated by the MTT method. The structures of the 5-hour biofilms were observed by using confocal laser scanning microscopy, and QS-related gene expressions were investigated by real-time PCR. The luxS mutant strain exhibited higher biofilm adherence and aggregation, but lower hydrophobicity than the wild-type strain. The confocal laser scanning microscopy images revealed that the wild-type strain tended to form smaller aggregates with uniform distribution, whereas the luxS mutant strain aggregated into distinct clusters easily discernible in the generated biofilm. Most of the genes examined were downregulated in the biofilms formed by the luxS mutant strain, except the gtfB gene. QS luxS gene can affect the initial biofilm formation by S. mutans.