Strain Chromobacterium Violaceum Research Articles

Characterization of N-Acylhomoserine Lactones Produced by Bacteria Isolated from Industrial Cooling Water Systems.

The cooling water systems are used to remove heat generated in the various industries. Biofouling of the cooling water systems causes blocking of condenser pipes and the heat exchanger tubes. In many Gram-negative bacteria, N-acylhomoserine lactone (AHL) are used as quorum-sensing signal molecule and associated with biofilm formation. To investigate the relationship between quorum sensing and biofouling in the cooling water system, we isolated a total of 192 bacterial strains from the five cooling water systems, and screened for AHL production. Seven isolates stimulated AHL-mediated purple pigment production in AHL reporter strain Chromobacterium violaceum CV026 or VIR07. Based on their 16S rRNA gene sequences, AHL-producing isolates were assigned to Aeromonas hydrophila, Lysobacter sp., Methylobacterium oryzae, and Bosea massiliensis. To the best of our knowledge, B. massiliensis and Lysobacter sp. have not been reported as AHL-producing species in the previous researches. AHLs extracted from the culture supernatants of B. massiliensis and Lysobacter sp. were identified by liquid chromatography-mass spectrometry. AHLs produced by B. massiliensis were assigned as N-hexanoyl-l-homoserine lactone (C6-HSL), N-(3-oxohexanoyl)-l-homoserine lactone (3-oxo-C6-HSL), and N-(3-oxooctanoyl)-l-homoserine lactone (3-oxo-C8-HSL). AHLs produced by Lysobacter sp. were assigned as N-decanoyl-l-homoserine lactone (C10-HSL) and N-(3-oxodecanoyl)-l-homoserine lactone (3-oxo-C10-HSL). This is the first report of identification of AHLs produced by B. massiliensis and Lysobacter sp. isolated from the cooling water system.

Tannin-Rich Fraction from Pomegranate Rind Inhibits Quorum Sensing in Chromobacterium violaceum and Biofilm Formation in Escherichia coli.

Pomegranate rind has been found to inhibit numerous pathogens, mostly attributed to its tannin fraction. The present study was conducted to investigate the quorum sensing (QS) inhibition effect of tannin-rich fraction from pomegranate rind (TFPR) by using an indicator strain Chromobacterium violaceum. Meanwhile, its effect on biofilm formation and motility of Escherichia coli was evaluated. It was shown that TFPR inhibited QS-regulated violacein pigment production. Biofilm formation and motility of E. coli were also hindered by TFPR. Transcriptional analysis further showed that TFPR repressed expressions of curli genes (csgB and csgD) and various motility genes (fimA, fimH, flhD, motB, qseB, and qseC). Our findings indicated that TFPR has potential application for controlling E. coli contaminations or biofilms in the food industry.

Anti-quorum sensing activity of spice oil nanoemulsions against food borne pathogens

Quorum sensing (QS) mechanism plays a vital role in many bacterial species, which is found to be implicated in various factors including bacterial pathogenicity and food spoilage. Therefore interrupting the QS mechanism may be an attractive strategy to develop novel QS-based anti-bacterial drugs. Here, we investigated the quorum sensing inhibitory activity of spice oil nanoemulsions (NE) and its antibiofilm property against selected food pathogens using the biosensor strain Chromobacterium violaceum CV026. Essential oil nanoemulsions were formulated by ultrasonic emulsification of Tween80 and water. The emulsification method for nanoformulations was standardized with Tween80 concentration and sonication time to attain high physical solidity and minimum droplet diameter. Stable cumin (Cuminum cyminum), pepper (Piper nigrum), and fennel (Foeniculum vulgare) oil nanoemulsions with respective droplet diameters of 52.89 nm, 82.08 nm and 59.52 nm were formulated by ultrasonic emulsification. Nanoemulsions at all tested concentrations inhibited the production of violacein pigment in C. violaceum CV026 below their minimum inhibitory concentration (MIC) level. In addition, the upshot of the current investigation reveals that all three nanoemulsions strongly interferes with quorum regulated phenotypes like biofilm formation and exopolysaccharide (EPS) production.Above findings lay a solid foundation for the utilization of nanoemulsions as QS-based antibacterial/anti-biofilm agents to manage food borne pathogens and biofilm formation in food industries.

A redox mediated UME biosensor using immobilized Chromobacterium violaceum strain R1 for rapid biochemical oxygen demand measurement

An effective ferricyanide-mediated microbial biochemical oxygen demand (BOD) biosensor was constructed and used for BOD determination in a water system. This BOD sensor uses ultramicroelectrode (UME) technology in which the tip of the sensor consists of a two-electrode system (10-μm Pt working electrode and Pt counter electrode). Because of their small size, UMEs have relatively large diffusion layers and small overall currents enabling rapid achievement of useful steady-state conditions with very high scan rates. Living Chromobacterium violaceum R1 cells (isolated from pineapple industry wastewater) were immobilized on the surface of the UME working electrode using a calcium alginate gel and further enclosure by a layer of polyamide membrane. Glucose-glutamic acid (GGA) solution was used as the standard solution. The amperometric measurement was optimized at +450mV operating potential and 30mM ferricyanide in a 0.1M phosphate buffer (pH 7.0) at 26°C. The sensor exhibited a linear response ranging from 20 to 225mg O2 L−1 BOD5 for standard GGA solution and 25 to 230mg O2 L−1 BOD5 for OECD synthetic wastewater with a response time of 30min. Repeatability and reproducibility of the biosensor were within the limits set by the APHA; i.e., less than 15.4%. The rapid BOD estimation of the biosensor is applicable for measuring samples with a high content of fast and easily assimilated compounds. When used to estimate the BOD of various wastewaters, the developed biosensor provided values comparable to those obtained using the conventional BOD5 method.

Inhibitory effect of marine cyanobacterial extract on biofilm formation and virulence factor production of bacterial pathogens causing vibriosis in aquaculture

Emergence of extended antibiotic resistance among bacterial pathogens often leads to the failure of existing antibiotics to treat bacterial infections; therefore, there is an urgent need to look for novel alternative treatment measures. The aim of this study was to evaluate the anti-quorum sensing (QS) potential of Synechococcus sp., to prevent the onset of bacterial infections as an alternate to antibiotics. A total of 110 marine cyanobacterial strains were screened for their anti-QS activity against biomarker strain Chromobacterium violaceum (ATCC 12472) and aquatic bacterial pathogens Vibrio harveyi (MTCC 3438) and Vibrio vulnificus (MTCC 1145). Of the 110 strains tested, the extract of unicellular algae Synechococcus sp. (Q-25) exhibited the efficient reduction in the production of violacein pigment of C. violaceum to the level of 82 %, bioluminescence of V. harveyi to 91 % and protease in V. vulnificus to 63 %. In V. harveyi and V. vulnificus, it exhibited a significant reduction of 71 and 84 % in biofilm formation and 66 and 68 % in EPS production, respectively, without any antibacterial activity. Confocal laser scanning microscopic and light microscopic analyses further confirmed that the Q-25 extract effectively prevented initial attachment as well as disrupting the architecture of mature biofilm, when compared to their untreated controls. In addition, the characterization of active principle by gas chromatography–mass spectrometry analysis confirmed the presence of stable bioactive compound hexadecanoic acid in the extract. Hence, this study clearly revealed the antibiofilm and QS inhibitory potential of the cyanobacterium, Synechococcus sp.

Effect of salinity and incubation time of planktonic cells on biofilm formation, motility, exoprotease production, and quorum sensing of Aeromonas hydrophila

The aim of this study was to determine the effect of salinity and age of cultures on quorum sensing, exoprotease production, and biofilm formation by Aeromonas hydrophila on stainless steel (SS) and crab shell as substrates. Biofilm formation was assessed at various salinities, from fresh (0%) to saline water (3.0%). For young and old cultures, planktonic cells were grown at 30 °C for 24 h and 96 h, respectively. Biofilm formation was assessed on SS, glass, and crab shell; viable counts were determined in R2A agar for SS and glass, but Aeromonas-selective media was used for crab shell samples to eliminate bacterial contamination. Exoprotease activity was assessed using a Fluoro™ protease assay kit. Quantification of acyl-homoserine lactone (AHL) was performed using the bioreporter strain Chromobacterium violaceum CV026 and the concentration was confirmed using high-performance liquid chromatography (HPLC). The concentration of autoinducer-2 (AI-2) was determined with Vibrio harveyi BB170. The biofilm structure at various salinities (0–3 %) was assessed using field emission electron microscopy (FESEM). Young cultures of A. hydrophila grown at 0–0.25% salinity showed gradual increasing of biofilm formation on SS, glass and crab shell; swarming and swimming motility; exoproteases production, AHL and AI-2 quorum sensing; while all these phenotypic characters reduced from 0.5 to 3.0% salinity. The FESEM images also showed that from 0 to 0.25% salinity stimulated formation of three-dimensional biofilm structures that also broke through the surface by utilizing the chitin surfaces of crab, while 3% salinity stimulated attachment only for young cultures. However, in marked contrast, salinity (0.1–3%) had no effect on the stimulation of biofilm formation or on phenotypic characters for old cultures. However, all concentrations reduced biofilm formation, motility, protease production and quorum sensing for old culture. Overall, 0–0.25% salinity enhanced biofilm formation and expression of quorum sensing regulatory genes in young cultures, whereas these responses were reduced when salinity was >0.25%. In old cultures, salinity at any concentrations (0.1–3%) induced stress in A. hydrophila. The present study provides insight into the ecology of A. hydrophila growing on fish and crustaceans such as shrimp and crabs in estuarine and seawater.

Inhibition of quorum sensing mediated biofilm development and virulence in uropathogens by Hyptis suaveolens

Bacterial urinary tract infections (UTIs) are the most common nosocomial infections, accounting for about 40% of all hospital-acquired infections. The bacterial spectrum of nosocomial UTIs is broad and the treatment of UTIs is becoming difficult owing to the emergence of drug resistance. Therefore, it is reasonable to investigate novel and alternative therapeutic strategies to treat UTIs. Since UTIs are caused by uropathogens with quorum sensing (QS)-dependent biofilm forming abilities, interruption of QS systems may be a novel approach to combat drug resistance. In the present study, a methanol extract (and hexane extract derived from it) of the medicinal plant Hyptis suaveolens (L.) were shown to have anti-QS activity against the biosensor strain Chromobacterium violaceum (ATCC 12472). Furthermore, the hexane extract of H. suaveolens (HEHS) inhibited biofilm formation by uropathogens such as Escherichia coli, Proteus vulgaris, Proteus mirabilis, Klebsiella pneumoniae and Serratia marcescens. HEHS promotes the loosening of biofilm architecture and strongly inhibits in vitro biofilm formation by uropathogens, which was more apparent from microscopic images. In addition to this, HEHS reduces the production of QS-dependent virulence factors like protease and hemolysin, along with motility. The partial purification and GC-MS analysis of the active fraction revealed the presence of several therapeutically important compounds which may synergistically act on the uropathogens and possibly reduce the QS-dependent phenotypes. These findings suggest HEHS as potential phytotherapeutic agent which can be employed to formulate protective strategies against biofilm linked infections caused by uropathogens.

Quorum quenching activity of Syzygium cumini (L.) Skeels and its anthocyanin malvidin against Klebsiella pneumoniae

Many bacterial species use their intercellular signaling mechanism called quorum sensing (QS), which is found to be implicated in various factors including bacterial pathogenicity and food spoilage. Interrupting the bacterial communication is an attractive strategy to develop novel QS-based antibacterial drugs. Present study is aimed to investigate the quorum sensing inhibitory activity of Syzygium cumini and its anti-biofilm property against opportunistic pathogen using a biosensor strain Chromobacterium violaceum CV026. Ethanol extract of S. cumini was investigated for its anti-QS activity, and the possible active component was identified by docking with LasR receptor protein. Based on docking analysis, methanol extract was enriched for its total anthocyanin (STA) and its effect on QS regulated phenotypes was assessed. STA specifically inhibited the violacein production in C. violaceum; biofilm formation and EPS production in Klebsiella pneumoniae up to 82, 79.94 and 64.29% respectively. Synergistic activity of conventional antibiotics with STA enhanced the susceptibility of K. pneumoniae up to 58.45%. Molecular docking analysis of active components attributes the QSI activity of S. cumini to malvidin. Malvidin exhibited highest ligand binding with LasR receptor protein with docking score more than −7. Effect of malvidin to interrupt the QS regulated phenotypes was also assessed, and it was found to reduce the violacein production, biofilm formation and EPS production of K. pneumoniae in a concentration-dependent manner. These findings suggest that S. cumini can be used as novel QS-based antibacterial/anti-biofilm agent to manage food-borne pathogens and to increase food safety.

Enzyme multilayer coatings inhibit Pseudomonas aeruginosa biofilm formation on urinary catheters.

Bacteria use a signaling mechanism called quorum sensing (QS) to form complex communities of surface-attached cells known as biofilms. This protective mode of growth allows them to resist antibiotic treatment and originates the majority of hospital-acquired infections. Emerging alternatives to control biofilm-associated infections and multidrug resistance development interfere with bacterial QS pathways, exerting less selective pressure on bacterial population. In this study, biologically stable coatings comprising the QS disrupting enzyme acylase were built on silicone urinary catheters using a layer-by-layer technique. This was achieved by the alternate deposition of negatively charged enzyme and positively charged polyethylenimine. The acylase-coated catheters efficiently quenched the QS in the biosensor strain Chromobacterium violaceum CECT 5999, demonstrated by approximately 50% inhibition of violacein production. These enzyme multilayer coatings significantly reduced the Pseudomonas aeruginosa ATCC 10145 biofilm formation under static and dynamic conditions in an in vitro catheterized bladder model. The quorum quenching enzyme coatings did not affect the viability of the human fibroblasts (BJ-5ta) over 7 days, corresponding to the extended useful life of urinary catheters. Such enzyme-based approach could be an alternative to the conventional antibiotic treatment for prevention of biofilm-associated urinary tract infections.

Marine-derived quorum-sensing inhibitory activities enhance the antibacterial efficacy of tobramycin against Pseudomonas aeruginosa.

Bacterial epiphytes isolated from marine eukaryotes were screened for the production of quorum sensing inhibitory compounds (QSIs). Marine isolate KS8, identified as a Pseudoalteromonas sp., was found to display strong quorum sensing inhibitory (QSI) activity against acyl homoserine lactone (AHL)-based reporter strains Chromobacterium violaceum ATCC 12472 and CV026. KS8 supernatant significantly reduced biofilm biomass during biofilm formation (−63%) and in pre-established, mature P. aeruginosa PAO1 biofilms (−33%). KS8 supernatant also caused a 0.97-log reduction (−89%) and a 2-log reduction (−99%) in PAO1 biofilm viable counts in the biofilm formation assay and the biofilm eradication assay respectively. The crude organic extract of KS8 had a minimum inhibitory concentration (MIC) of 2 mg/mL against PAO1 but no minimum bactericidal concentration (MBC) was observed over the concentration range tested (MBC > 16 mg/mL). Sub-MIC concentrations (1 mg/mL) of KS8 crude organic extract significantly reduced the quorum sensing (QS)-dependent production of both pyoverdin and pyocyanin in P. aeruginosa PAO1 without affecting growth. A combinatorial approach using tobramycin and the crude organic extract at 1 mg/mL against planktonic P. aeruginosa PAO1 was found to increase the efficacy of tobramycin ten-fold, decreasing the MIC from 0.75 to 0.075 µg/mL. These data support the validity of approaches combining conventional antibiotic therapy with non-antibiotic compounds to improve the efficacy of current treatments.

Identification and analysis of the salt tolerant property of AHL lactonase (AiiATSAWB) of Bacillus species

Bacterial biofilms communicate by a process called Quorum Sensing. Gram negative bacterial pathogens specifically talk through the production, detection, and response to the signal or autoinducer called Acyl Homoserine Lactones. Bacterial lactonases are important AHL hydrolysing or quorum quenching enzymes. The present study deals with ten endospore forming gram positive isolates of the saltern soil. Preliminary screening for Quorum Quenching activity with the QS Inhibition indicator strain Chromobacterium violaceum ATCC 12472, showed positive activity in four isolates namely TS2, TS16, TSAWB, and TS53B. AHL lactonase (AiiA) specific primers amplified Acyl Homoserine Lactone lactonase gene in the TSAWB genome alone. Phylogenetic relationship of the identified AiiATSAWB confirmed its evolutionary relationship with bacterial AiiA like AHL lactonase of the metallo-beta-lactamase super family. Our in vitro AHL hydrolysis assay under wide percentage (0-5) of salt solutions with TSAWB isolate and also its intracellular soluble protein fraction showed halotolerant AHL hydrolysis ability of the AiiATSAWB enzyme. In silico determination of putative tertiary structure, the ESBRI derived conserved salt bridges, aminoacid residue characterization with high mole percent of acidic and hydrophobic residues reaffirmed the halotolerant ability of the enzyme. So we propound the future use of purified AiiATSAWB , as hypertonic suspension for inhalation to substitute the action of inactivated host's paraoxonase in treating Pseudomonas aeruginosa infection in cystic fibrosis patients.

Characterization of Chromobacterium violaceum pigment through a hyperspectral imaging system

In this paper, a comprehensive spatio-spectral and temporal analysis for Chromobacterium violaceum colonies is reported. A hyperspectral imaging (HSI) system is used to recover the spectral signatures of pigment production in a non-homogeneous media with high spectral resolution and high sensitivity in vivo, without destructing the sample. This non-contact sensing technique opens avenues to study the temporal growing of a specific section in the bacterial colony. Further, from a 580 [nm] and 764 [nm] spatio-spectral time series, a wild-type and mutant Chromobacterium violaceum strains are characterized. Such study provides quantitative information about kinetic parameters of pigment production and bacterial growing.

Production and Degradation ofN-Acylhomoserine Lactone Quorum Sensing Signal Molecules in Bacteria Isolated from Activated Sludge

N-Acylhomoserine lactones (AHLs) function as quorum-sensing signaling molecules in many Gram-negative bacteria. We isolated a total of 672 bacterial strains from activated sludge obtained from seven sewage treatment plants in Tochigi Prefecture, Japan, and screened for AHL-producing and degrading strains. Isolates (n = 107) stimulated AHL-mediated purple pigment production in AHL reporter strains Chromobacterium violaceum CV026 and VIR07. Based on their 16S rRNA gene sequences, most of these AHL-producing isolates were assigned to the genus Aeromonas, and they were divided into six groups. Isolates (n = 46) degraded N-decanoyl-L-homoserine lactone (C10-HSL) within 24 h. Based on their 16S rRNA gene sequences, the most dominant AHL-degrading isolates were assigned to the genus Acinetobacter and divided into six groups. Strains Ooi24, Omo91, and Uzu81, which showed higher C10-HSL-degrading activity, showed putative AHL-acylase activity.

Vestigialization of arsenic resistance phenotypes/genotypes in Chromobacterium violaceum strains thriving in pristine Brazilian sites

The performance of the arsenic resistance arsRCB operon of the ubiquitous bacterium Chromobacterium violaceum has been employed as a descriptor of the genomic persistence of traits for long-ceased environmental conditions. To this end, a reliable reference for arsenic resistance/sensitivity was created in the C. violaceum ATCC 12472-type strain by deleting its genomic ars operon. This allowed tracing all of the arsenic resistance of this bacterium to the ars gene cluster. A sample of 15 environmental C. violaceum strains isolated from pristine Amazonian sites (both rain forest and savannah) were then examined for the presence of arsRCB amplicons, for arsenic resistance, and for indications of decay of ars genes into pseudogenes. While the sites of origin of such strains have no record of industrial or geological exposure to arsenic, all of them bore the standard arsRCB cluster in their genome, but they did differ in their resistance to the metalloid. This ranged from complete sensitivity to tolerance levels above the reference C. violaceum ATCC12472. Inspection of relevant DNA sequences, phenotypic analyses and monitoring expression profiles of the ars genes in each strain exposed a distinct genetic drift of the system toward vestigialization, that is, accumulation of mutations leading to the loss of a non-used character. In particular, quantitative PCR revealed inducibility by arsenic to be the first trait to be lost in the route toward decaying and eventual nonfunctionalization. The data presented fit evolutionary predictions on elimination of non-used characters through a mere genetic drift accumulating neutral mutations.

Enhancing gold recovery from electronic waste via lixiviant metabolic engineering in Chromobacterium violaceum

Conventional leaching (extraction) methods for gold recovery from electronic waste involve the use of strong acids and pose considerable threat to the environment. The alternative use of bioleaching microbes for gold recovery is non-pollutive and relies on the secretion of a lixiviant or (bio)chemical such as cyanide for extraction of gold from electronic waste. However, widespread industrial use of bioleaching microbes has been constrained by the limited cyanogenic capabilities of lixiviant-producing microorganisms such as Chromobacterium violaceum. Here we show the construction of a metabolically-engineered strain of Chromobacterium violaceum that produces more (70%) cyanide lixiviant and recovers more than twice as much gold from electronic waste compared to wild-type bacteria. Comparative proteome analyses suggested the possibility of further enhancement in cyanogenesis through subsequent metabolic engineering. Our results demonstrated the utility of lixiviant metabolic engineering in the construction of enhanced bioleaching microbes for the bioleaching of precious metals from electronic waste.

Matrix-assisted laser desorption ionisation–time of flight mass spectrometry for rapid identification of Laribacter hongkongensis

Laribacter hongkongensis is a Gram-negative, facultative anaerobic, motile, S-shaped, urease-positive bacillus associated with invasive infections in liver cirrhosis patients and community-acquired gastroenteritis. Most cases of L hongkongensis infections occur in...

F18ab Escherichia coli flagella expression is regulated by acyl-homoserine lactone and contributes to bacterial virulence

To investigate the effect of the Quorum Sensing (QS)-I system on the expression of virulence factors in Shiga toxin producing and verotoxin-producing Escherichia coli (STEC and VTEC), the yenI gene from Yersinia enterocolitica was cloned into E. coli F18ab 107/86. Recombinant E. coli transformed with yenI produced acyl-homoserine lactone synthase (AHL), as measured using cross-streaking assays with the reporter biosensor strain Chromobacterium violaceum CV026. The AI-1 positive recombinant F18ab E. coli exhibited impaired expression of flagella, decreased motility, reduced biofilm formation and AI-2 production, as well as attenuated adherence and invasion on IPEC-J2 cells. This study provides new insights to the crucial function of AI-1 in regulating STEC virulence.

Rose, Clove, Chamomile Essential Oils and Pine Turpentine Inhibit Quorum Sensing in Chromobacterium violaceum and Pseudomonas aeruginosa

Forty-two natural essential and edible oils were tested for their ability to inhibit bacterial QS to create a comprehensive database of candidate inhibitor compounds which may provide significant option for treatment of bacterial infections. QS inhibitory activities of oils were screened using biomonitor organisms; Quorum Sensing Inhibitor Selector strain1 (QSIS1) and Chromobacterium violaceum strain CV026, C. violaceum strain VIR07, C. violaceum strain ATCC 12472 and opportunistic pathogen Pseudomonas aeruginosa PAO1, clinical isolate of P. aeruginosa (PCI). While eleven of tested oils were found to be positive for QSIS1; Rose, clove, chamomile essential oils and pine turpentine exhibited significant decrease in violacein production for Chromobacterium strains tested at very low concentrations and showed anti-biofilm activity in clinical isolate of P. aeruginosa (PCI). The chemical compositions of rose, clove, chamomile essential oils and pine turpentine were determined by GC-MS. Results of this study showed that there are noteworthy QSI compounds in the rose, clove, chamomile essential oils and pine turpentine and this potential could be important treatment strategy for Pseudomonal infections.

Crystalline Xylitol Production by a Novel Yeast, Pichia caribbica (HQ222812), and Its Application for Quorum Sensing Inhibition in Gram-Negative Marker Strain Chromobacterium violaceum CV026

Xylitol, a sugar alcohol, is fast gaining ground over other artificial sugar substitutes owing to its advantageous properties. Xylitol is a safer alternative for diabetics because of insulin-independent metabolism. It has beneficial properties suitable to form an important part of odontological formulations. Conventional commercial production of xylitol involves harsh chemical method operating at high temperature and pressure. Thus, microbial production of xylitol is preferred over chemical method, and yeasts have been extensively exploited for this purpose. In the present manuscript, quantitative production of xylitol from D-xylose with the yield of 0.852gm/gm and volumetric productivity of 1.83gm/l/h in crystalline form, using novel yeast Pichia caribbica is reported. Also, a mild, safe procedure for product extraction is described. The ability of xylitol to act as a quorum sensing antagonist in gram-negative marker strain Chromobacterium violaceum CV026 has been demonstrated for the first time.

Attenuation of Pseudomonas aeruginosa virulence by marine invertebrate-derived Streptomyces sp.

The study aimed to discover quorum sensing (QS) inhibitors from marine sponge-derived actinomycetes and analyse its inhibitory activities against QS-mediated virulence factors in Pseudomonas aeruginosa. Seventy-two actinomycetes isolated from marine invertebrates collected from the western coast of India were screened against the QS indicator strain Chromobacterium violaceum CV12472. Methanol extracts of 12 actinomycetes showing inhibition of violacein production were accessed for downregulation of QS-mediated virulence factors like swarming, biofilm formation, pyocyanin, rhamnolipid and LasA production in Ps. aeruginosa ATCC 27853. The isolates NIO 10068, NIO 10058 and NIO 10090 exhibited very good anti-QS activity, with NIO 10068 being the most promising one. Mass spectrometric analysis of NIO 10068 methanol extract revealed the presence of cinnamic acid and linear dipeptides proline-glycine and N-amido-α-proline in the active extract. Detailed investigation suggested that although linear dipeptide Pro-Gly is to some extent responsible for the observed biological activity, cinnamic acid seems to be the main compound responsible for it. Marine-derived actinomycetes are a potential storehouse for QS inhibitors. This is the first report not only on marine sponge-associated Streptomyces for anti-QS in Ps. aeruginosa but also on cinnamic acid and proline-derived linear dipeptides proline-glycine as QS inhibitors. The results reveal that marine-derived actinomycetes may not only play a role in the defensive mechanism of their host but also lead to new molecules useful in the development of novel antivirulence drugs.