Chromobacterium Violaceum Research Articles

A widespread response of Gram-negative bacterial acyl-homoserine lactone receptors to Gram-positive Streptomyces γ-butyrolactone signaling molecules.

Cell-cell communication is critical for bacterial survival in natural habitats, in which miscellaneous regulatory networks are encompassed. However, elucidating the interaction networks of a microbial community has been hindered by the population complexity. This study reveals that γ-butyrolactone (GBL) molecules from Streptomyces species, the major antibiotic producers, can directly bind to the acyl-homoserine lactone (AHL) receptor of Chromobacterium violaceum and influence violacein production controlled by the quorum sensing (QS) system. Subsequently, the widespread responses of more Gram-negative bacterial AHL receptors to Gram-positive Streptomyces signaling molecules are unveiled. Based on the cross-talk between GBL and AHL signaling systems, combinatorial regulatory circuits (CRC) are designed and proved to be workable in Escherichia coli (E. coli). It is significant that the QS systems of Gram-positive and Gram-negative bacteria can be bridged via native Streptomyces signaling molecules. These findings pave a new path for unlocking the comprehensive cell-cell communications in microbial communities and facilitate the exploitation of innovative regulatory elements for synthetic biology.

Could the analgesic drugs, paracetamol and indomethacin, function as quorum sensing inhibitors?

The current failure of antimicrobials in treating life-threatening diseases, the high rate of multidrug resistant pathogens and the slow progress in the development of new antibiotics directed scientists to develop antivirulence drugs that targets quorum sensing (QS). In many microbes, QS acts as a communication system which control pathogenicity of microbes. Analgesics can be beneficial in controlling virulence traits of microbes and hence they may augment the efficacy of antimicrobials.In this study, two analgesics were screened for the inhibition of QS in Chromobacterium violaceum CV026 and their effects on virulence production in Pseudomonas aeruginosa PAO1 strain and clinical isolates of Acinetobacter baumannii were evaluated. The traits investigated were biofilm formation, pyocyanin and rhamnolipid production, twitching, swarming or surface associated motilities, production of protease, phospholipase and gelatinase enzymes and sensitivity to oxidative stress. Relative expression of abaI gene was calculated by performing qRT-PCR. Docking analysis of paracetamol as QSI (quorum sensing inhibitor) of AbaI and AbaR proteins was performed.Paracetamol inhibited QS in CV026, but indomethacin devoids anti-QS activity. Paracetamol inhibited virulence factors of PAO1. It strongly inhibited biofilm formation, and swarming by 66.4% and 57.1%, respectively. While, it moderately to slightly inhibited rhamnolipid, pyocyanin, gelatinase, resistance to oxidative stress, protease and twitching motility by 33.3%, 33.1% 17.5%, 9.1%, 8.7% and 7.7%, respectively. For A. baumannii, paracetamol strongly inhibited biofilm by 39.7–93% and phospholipase enzyme by 8.7–100%, reduced twitching and surface motility by 6.7–82.5% and 7.7–29.4%, respectively, And slightly reduced sensitivity to oxidative stress by 3.3–36.4%. Paracetamol at sub-MIC suppressed the expression of abaI gene by 32% in A. baumannii. Docking studies suggested that paracetamol can bind to AbaR and AbaI proteins and bind more to AbaR, hence it may act by inhibiting AHL signal reception.As a conclusion, paracetamol, beside its analgesic activity, has anti-QS activity and could be used in the eradication of P. aeruginosa and A. baumannii infections in combination with antibiotics.

Interference of quorum sensing regulated bacterial virulence factors and biofilms by Plumbago zeylanica extract.

There has been tremendous spread of antimicrobial resistance globally, mainly due to the excessive and unnecessary use of antibiotics, making the situation alarming. This has created a need for the development of alternative strategies to selectively target the bacterial pathogenicity without exerting selection pressure for the development of antimicrobial resistance. Targeting quorum sensing (QS)-mediated virulence and biofilms by nontoxic natural products is gaining importance as new control strategy to combat the virulence and biofilms of pathogenic bacteria. In this study, the crude extract of Plumbago zeylanica was fractioned in different solvents using liquid-liquid partitioning to obtain the most bioactive fraction. The inhibitory effect of the bioactive extract of P. zeylanica on QS at sub-minimum inhibitory concentrations (MICs) was studied against Chromobacterium violaceum 12472, Pseudomonas aeruginosa PAO1, and Serratia marcescens MTCC 97. Biofilm inhibition was studied using microtiter plate assay, scanning electron microscopy, and confocal laser scanning microscopy. Major phytocompounds detected were cinnamaldehyde dimethyl acetal, plumbagin, asarone, 4-chromanol, phthalic acid, palmitic acid, ergost-5-en-3-ol, stigmasterol, and β-sitosterol. The violacein production in C. violaceum 12472 was reduced by >80% in the presence of P. zeylanica hexane fraction (PZHF; 200 μg/ml). The most active PZHF inhibited QS-mediated virulence factors of P. aeruginosa PAO1 such as pyocyanin, pyoverdin, rhamnolipid production, motility, etc., significantly at sub-MICs. Similarly, PZHF showed 59 to 76% inhibition of biofilm formation of above test pathogens. The findings revealed that active fraction of P. zeylanica was effective against the QS-regulated functions and biofilms development of Gram -ve pathogenic bacteria.

Coumarin Exhibits Broad-Spectrum Antibiofilm and Antiquorum Sensing Activity against Gram-Negative Bacteria: In Vitro and In Silico Investigation.

Quorum sensing (QS) and biofilm inhibition are recognized as the novel drug targets for the broad-spectrum anti-infective strategy to combat the infections caused by drug-resistant bacterial pathogens. Many compounds from medicinal plants have been found to demonstrate anti-infective activity. However, broad-spectrum anti-QS and antibiofilm efficacy and their mode of action are poorly studied. In this study, the efficacy of coumarin was tested against QS-regulated virulent traits of Gram-negative bacteria. Coumarin inhibited the production of violacein pigment in Chromobacterium violaceum 12472 by 64.21%. Similarly, there was 87.25, 70.05, 76.07, 58.64, 48.94, and 81.20% inhibition of pyocyanin, pyoverdin, and proteolytic activity, lasB elastase activity, swimming motility, and rhamnolipid production, respectively, in Pseudomonas aeruginosa PAO1. All tested virulence factors of Serratia marcescens MTCC 97 were also suppressed by more than 50% at the highest sub-minimum inhibitory concentration. Moreover, the biofilms of bacterial pathogens were also inhibited in a dose-dependent manner. Molecular docking and molecular dynamics (MD) simulation gave insights into the possible mode of action. The binding energy obtained by docking studies ranged from −5.7 to −8.1 kcal mol–1. Coumarin was found to be docked in the active site of acylhomoserine lactone (AHL) synthases and regulatory proteins of QS. MD simulations further supported the in vitro studies where coumarin formed a stable complex with the tested proteins. The secondary structure of all proteins showed a negligible change in the presence of coumarin. Computational studies showed that the possible mechanisms of anti-QS activity were the inhibition of AHL synthesis, antagonization of QS-regulatory proteins, and blocking of the receptor proteins. The findings of this study clearly highlight the potency of coumarin against the virulence factors of Gram-negative bacterial pathogens that may be developed as an effective inhibitor of QS and biofilms.

Physicochemical Composition and Characterization of Bioactive Compounds of Mulberry (Morus indica L.) Fruit During Ontogeny.

Mulberry fruit is well recognized as one of the richest sources of bioactive compounds. We investigated the physicochemical composition and characterized the bioactive compounds during different ripening stages of mulberry (Morus indica) fruit and evaluated their anti-quorum sensing activity on Chromobacterium violaceum. The proximate components such as carbohydrates, proteins and lipids were found to be high in the ripe fruit compared to unripe and mid-ripe fruit. The ripe fruit contained higher content of total phenolics and flavonoids (336.05 and 282.55mg/100g fresh weight (FW), respectively). Epicatechin and resveratrol were the major polyphenols detected in the fruit with the range 5.13-19.46 and 4.07-14.45mg/100g FW, respectively. Chlorogenic acid and myricetin were predominant in the unripe and mid-ripe fruit (7.14 and 1.84mg/100g FW, respectively). The fruit was found to be an excellent source of anti-diabetic compound 1-deoxynojirimycin. The highest content of 1-deoxynojirimycin was present in the mid-ripe fruit, with a content of 2.91mg/100g FW. Furthermore, fruit extracts exhibited anti-quorum sensing activity against Chromobacterium violaceum by effectively inhibiting violacein production. Ripe fruit extracts showed the highest activity of 76.30% at 1mg/mL and thus, could be used as a potent anti-quorum sensing agent. The results could be promising in the selection of appropriate developmental stages for M. indica fruit commercial exploitation inthe food formulations rich in potential health components.

Bioleaching of metals from waste printed circuit boards using bacterial isolates native to abandoned gold mine.

In the present study, native bacterial strains isolated from abandoned gold mine and Chromobacterium violaceum (MTCC-2656) were applied for bioleaching of metals from waste printed circuit boards (WPCBs). Toxicity assessment and dose-response analysis of WPCBs showed EC50 values of 128.9, 98.7, and 90.8g/L for Bacillus sp. SAG3, Bacillus megaterium SAG1 and Lysinibacillus sphaericus SAG2, respectively, whereas, for C. violaceum EC50 was 83.70g/L. This indicates the viable operation range and technological feasibility of metals bioleaching from WPCBs using mine isolates. The influencing factors such as pH, pulp density, temperature, and precursor molecule (glycine) were optimized by one-factor at a time method (OFAT). The maximum metal recovery occurred at an initial pH of 9.0, a pulp density of 10g/L, a temperature of 30°C and a glycine concentration of 5g/L, except for L. sphaericus which showed optimum activity at initial pH of 8.0. Under optimal conditions the metals recovery of Cu and Au from WPCBs were recorded as 87.5 ± 8% and 73.6 ± 3% for C. violaceum and 72.7 ± 5% and 66.6 ± 6% for B. megaterium, respectively. Kinetic modeling results showed that the data was best described by first order reaction kinetics, where the rate of metal solubilization from WPCBs depended upon microbial lixiviant production. This is the first report on bioleaching of metals from e-waste using bacterial isolates from the gold mine of Solan, HP. Our study demonstrated the potential of bioleaching for resource recovery from WPCBs dust, aimed to be disposed at landfills, and its effectiveness in extraction of elements those are at high supply risk and demand.

Chromobacterium violaceum: A Review of an Unexpected Scourge.

Chromobacterium violaceum is a common environmental bacterium that rarely causes disease in humans but has a high fatality rate if it does. Due to the rarity of the cases, clinicians are often unaware of the rapid progression of C. violaceum infection and its unexpected antibiotic resistance pattern, which contribute to the failure of patient management. Our review provides the clinical characteristics, possible sources of exposure, and comorbidities and determines factors associated with survival. We gathered information on 132 cases of C. violaceum causing disease in humans published between 1953 and 2020. Patients were predominantly male with a median age of 17.5, interquartile range (IQR) of 5.0–40.0 years, and a third of them were known to have immune deficiencies or comorbidities. Portals of entry were mainly through a wound in the leg and feet (28.0%), the torso (8.5%), or hands and arms (6.8%). It is not uncommon to acquire infection through unintended contact with contaminated water or dust through the mouth or inhalation. The median incubation period is 4.0 days (IQR 2.0–8.0 days) with a duration of clinical course of 17.5 days (IQR 8.0–30.8 days). The high rate of positive blood cultures (56.1%) and abscesses in internal organs (36.4%) shows the significant severity of this disease. Sepsis and Bacteremia were related to mortality with a risk ratio (RR) of 5.20 (95% CI, 0.831–32.58) and 2.14 (95% CI, 1.05–4.36), respectively. Appropriate antibiotic use prevented death at a RR 0.33 (95% CI, 0.21–0.52). Most patients who recovered and survived were treated with aminoglycosides, fluoroquinolones and carbapenems. This review shows the malignant nature of C. violaceum infection and the need for clinicians to be aware and provide prompt source management for patients. Appropriate empiric and targeted antibiotic regiment guided by susceptibility test results is of vital importance.

Plant growth promoting bacteria induce anti-quorum-sensing substances in chickpea legume seedling bioassay.

The online version contains supplementary material available at 10.1007/s12298-021-01034-x.

Assessment of antifungal potentials of violacein extract from Chromobacterium violaceum isolated from domestic and recreational water sources in Owerri, Imo State, Nigeria

Anti-Microbial Resistance (AMR) in superficial fungal infections are major worldwide public health problem that affects a large part of the human population globally. The antifungal potentials of violacein extracted from Chromobacterium violaceum isolated from domestic and recreational water sources in Owerri, Imo State, Nigeria, was assessed. Three water samples were collected from different locations of the Otamiri River, five from different swimming pools and three from different borehole locations in Owerri Metropolis. The samples were cultured on nutrient agar by pour plate method. The violet colonies of Chromobacterium violaceum were counted, characterized and identified. Water sample from Otamiri River station-1 had the highest bacteria count (20.00 × 101 CFU/Ml and 19.50 × 101 CFU/mL) respectively. Swimming pool 1 and 3 bacterial counts were (14.50 × 101 CFU/mL, 11.00 × 101 CFU/mL and 11.50 × 101 CFU/mL) respectively. For borehole 1, 2 and 3, swimming pool 2, 4 and 5 counts were (0.00 × 101 CFU/mL). The ethanolic extracts from the isolates (violacein) and the control drug (fluconazole 50µg/mL) both had inhibitory effects on the test organisms (Candida albicans and Aspergillus niger) at different concentrations. The MIC of Fluconazole on Candida albicans and A. niger were 25µg/ml and 50µg/Ml, respectively. Violacein from both swimming pools and Otamiri River isolates had MIC of 8.75mg/ml on A. niger and MIC of 4.375mg/ml on Candida albicans. Violacein which proved to have inhibitory effects on Candida albicans and Aspergillus niger can be harnessed for treatment of infections caused by these fungi.

Assessment of antifungal potentials of violacein extract from Chromobacterium violaceum isolated from domestic and recreational water sources in Owerri, Imo State, Nigeria

Assessment of antifungal potentials of violacein extract from Chromobacterium violaceum isolated from domestic and recreational water sources in Owerri, Imo State, Nigeria

Quorum sensing inhibitory effect of bergamot oil and aspidosperma extract against Chromobacterium violaceum and Pseudomonas aeruginosa.

Quorum sensing (QS) represents a major target for reducing bacterial pathogenicity and antibiotic resistance. This study identifies bergamot and aspidosperma as new potential sources of anti-QS agents. We investigated the anti-QS activity of plant materials on both Chromobacterium violaceum and Pseudomonas aeruginosa. Initially, we determined the minimum inhibitory concentrations (MICs) of plant materials using a broth microdilution method. Subsequently, we tested the effect of sub-MIC concentrations on QS-regulated traits and virulence factors production in test bacteria. Results revealed that bergamot and aspidosperma inhibited the ability of C. violaceum to produce violacein. Other QS-controlled phenotypes of C. violaceum, namely chitinolytic activity, motility, and biofilm formation, were also reduced by both plant materials. Moreover, QS-linked traits of P. aeruginosa were also reduced. Bergamot inhibited swarming but not swimming motility, while aspidosperma diminished both motility types in P. aeruginosa. Both plant materials also demonstrated antibiofilm activity and inhibited the production of protease and pyocyanin in P. aeruginosa. Furthermore, we tested the anti-QS effect of plant materials on the transcriptional level using RT-qPCR. Bergamot dramatically downregulated the C. violaceum autoinducer synthase gene cviI and the vioB gene involved in violacein biosynthesis, confirming the phenotypic observation on its anti-QS activity. Aspidosperma also reduced the expression of cviI and vioB but less drastically than bergamot. In P. aeruginosa, downregulation in the transcripts of the QS genes lasI, lasR, rhlI, and rhlR was also achieved by bergamot and aspidosperma. Therefore, data in the present study suggest the usefulness of bergamot and aspidosperma as sources of antivirulence agents.

Discovery of novel β-turn mimetic-based peptides as novel quorum sensing inhibitors of gram-negative bacteria

To date, a very limited number of peptides reported as quorum sensing inhibitors. Herein, we report the synthesis and evaluation of a series of β-turn mimetic-based peptides as potent quorum sensing inhibitors and antibiofilm formation. In this series, peptides P1, P4, and P5 showed very promising anti-quorum sensing activity on lasB-gfp reporter strain of Pseudomonas aeruginosa without affecting bacterial growth. Under our condition, these compounds also showed good anti-violacein production of Chromobacterium violaceum. In terms of antibiofilm formation, except P5, two β-turn mimetic-based peptides P1 and P4 showed maximum inhibition of 80% total biomass of Pseudomonas aeruginosa. This report provides the first β-turn mimetic-based scaffold for future drug development.

Vibrio alginolyticus influences quorum sensing-controlled phenotypes of acute hepatopancreatic necrosis disease-causing Vibrio parahaemolyticus.

BackgroundAcute hepatopancreatic necrosis syndrome (AHPND) caused by Vibrio parahaemolyticus strain (VPAHPND) impacts the shrimp industry worldwide. With the increasing problem of antibiotic abuse, studies on quorum sensing (QS) system and anti-QS compounds bring potential breakthroughs for disease prevention and treatment.MethodsIn this study, the cell-free culture supernatant (CFCS) and its extract of V. alginolyticus BC25 were investigated for anti-QS activity against a reporter bacteria, Chromobacterium violaceum DMST46846. The effects of CFCS and/ or extract on motility, biofilm formation and extracellular polymeric substances (EPSs) of VPAHPND PSU5591 were evaluated. Moreover, the effects of V. alginolyticus BC25 on virulence of VPAHPND PSU5591 were investigated by shrimp challenge test. The potentially active anti-QS compounds presented in the extract and effect on gene expression of VPAHPND PSU5591 were identified.ResultsThe CFCS of V. alginolyticus BC25 and its extract showed a significant anti-QS activity against the reporter bacteria as well as swimming and swarming motilities, biofilms, and EPSs production by VPAHPND PSU5591. Transcriptome analysis revealed that V. alginolyticus BC25 extract significantly reduced the flagella genes involved in biofilm formation and iron-controlled virulence regulatory gene of VPAHPND PSU5591. Whereas, the LuxR family transcriptional regulator gene, c-factor, a cell-cell signaling gene, and capsular polysaccharide were up-regulated. The potentially active anti-QS compounds identified in extract were Cyclo-(L-Leu-L-Pro), and Cyclo-(L-Phe-L-Pro). Furthermore, V. alginolyticus BC25 enhanced disease resistance against VPAHPND PSU5591 in tested shrimp larvae.ConclusionThese findings suggest that V. alginolyticus BC25 could provide natural anti-QS and anti-biofilms compounds and has great ability to be used as biocontrol agent against VPAHPND infection in shrimp aquaculture.

Anti-Quorum Sensing and Antioxidant Activity of Essential Oils Extracted From Juniperus Species, Growing Spontaneously in Tebessa Region (East of Algeria)

The chemical composition of essential oils (EOs) extracted from the aerial parts of 2 species of Juniperus was determined by Gas Chromatography-Mass Spectrometry (GC-MS). In total, 65 and 58 compounds accounting for 90.3% and 89.8% of the whole chemical composition of Juniperus oxycedrus (JO) and Juniperus phoenicea (JP) were identified, respectively, with α-pinene, α-amorphene, terpinen-4-ol, α-terpinene, and β-elemene, as major components. For the first time, the capacity to inhibit quorum-sensing for Chromobacterium violaceum CV026 and CV12472 by the investigated EOs was evaluated. Both oils exhibited good violacein inhibition on CV12472 with 100.0 ± 0.0% inhibition at minimal inhibition concentration (MIC) values. Besides, the quorum-sensing inhibition of CV026 was high at MIC for JO essential oil from fruits (JOF, 16.3 ± 2.0 mm), JO leaves (JOL, 12.5 ± 3.5 mm), JP fruits (JPF, 19.7 ± 2.5 mm), and JP leaves (JPL, 21.1 ± 5.0 mm). On both CV12472 and CV026, essential oil from J. phoenicea leaves was the most active inhibitor. All investigated EOs inhibited swarming motilities in flagellated Pseudomonas aeruginosa (PA01) in a concentration-dependent manner, and those from JP were more active than EOs from JO. Moreover, these EOs showed good antioxidant potential according to DPPH● and FRAP methods, especially the EO from JO leaves with an IC50 DPPH● inhibition value of 20.2 ± 1.0 mg/mL. Based on the obtained results, the investigated EOs are good candidates to combat microbial resistance be used as alternatives to conventional antibiotics, and equally find applications in food biosafety as preservatives.

Successful treatment of a patient with recurrent infection of Chromobacterium violaceum

BackgroundChromobacterium violaceum (C. violaceum) is a Gram-negative saprophytic bacterium that is widespread in tropical and subtropical environments, and belongs to conditional pathogenic bacteria. Human infection with C. violaceum is rare, and this can be fatal when the diagnosis and treatment are delayed, especially recurrent infection patients. Since clinicians lack the knowledge for C. violaceum, rapid diagnosis and early appropriate antimicrobial treatment remains challenging.Case presentationA 15-year-old male student was hospitalized for dark abscess, pustules, severe pain in both legs, and fever for 11 days. There were pustules with gray-white pus and red infiltrating plaques on the back, and the subcutaneous nodules could be touched in front of both tibias, with scab, rupture and necrotic tissue of the lower limb. The patient’s condition rapidly progressed. Therefore, next-generation sequencing (NGS), pustular secretion and blood culture were concurrently performed. The final diagnosis for this patient was C. violaceum infection by NGS. However, no bacterial or fungal growth was observed in the pustular secretion and blood culture. After 4 weeks of treatment, the patient was discharged from the hospital without any complications associated with C. violaceum infection.ConclusionRapid diagnosis and early appropriate antimicrobial treatment is the key to the successful treatment of C. violaceum infection, especially in patients with sepsis symptoms. This case highlights that NGS is a promising tool for the rapid diagnosis of C. violaceum infection, preventing the delayed diagnosis and misdiagnosis of C. violaceum infection in patients who tested negative for pustular secretion and blood culture.

Clinical and demographic pattern of chronic granulomatous disease (CGD) from a multicenter perspective: Malaysia\u2019s experience over 26 years

BackgroundA retrospective review of clinical manifestations and demographic pattern of patients diagnosed as chronic granulomatous disease (CGD) from 7 hospitals in Malaysia. An analysis of the available database would establish clinical characteristics, diagnoses and outcome including microbiologic pattern. Studying the demography allows us to document the occurrence of CGD amongst multiethnic groups and its geographical distribution for Malaysia.MethodsData from the Malaysia Primary Immunodeficiency Network (MyPIN) with cases of CGD diagnosed from 1991 until 2016 were collated and analysed.ResultsTwenty patients were diagnosed as CGD. Males (N = 13, 65%) outnumber females (N = 7, 35%). CGD is commonest amongst the Malays (65%) followed by the Chinese (15.0%), Indians (10.0%) and natives of Borneo (10.0%), reflecting the ethnic composition of the country. The mean age of diagnosis was 3.7 years. There was a positive family history in 40% of the cases. Abscess was the main presenting feature in 16 patients (80%) with one involving the brain. Pneumonia occurred in 10 (50%) and one with complicated bronchiectasis. Catalase-positive bacteria were the most commonly isolated pathogen with Chromobacterium violaceum predominating (N = 5, 25%) with consequent high mortality (N = 4, 80%). All CGD patients with C. violaceum infection displayed CD4 + (T helper cells) lymphopenia.ConclusionThis study has shown CGD occurs in the major ethnic groups of Malaysia. To the best of our knowledge, this is the first and the largest series of chronic granulomatous disease in South East Asia which may be reflective of similar clinical pattern in the region. C. violaceum infection is associated with a higher mortality in CGD patients in Malaysia. All the CGD patients with C. violaceum infection in this patient series displayed CD4 + (T helper) lymphopenia. We recorded rare clinical manifestation of CGD viz. brain abscess and bronchiectasis.

Clitoria ternatea ethanolic extract prevents dental caries via inhibiting Streptococcus mutans growth and quorum sensing

Our previous study demonstrated that the blue butterfly pea flower (Clitoria ternatea) had several pharmacological effects to treat inflammatory-related diseases, including edema and diabetes. However, its benefit for preventing dental caries and protecting the tooth has not been explored yet. Here, we investigated whether C. ternatea ethanolic extract (CTEE) prevented dental caries through antibacterial and antiquorum sensing activities toward oral pathogen Streptococcus mutans in vitro. CTEE was made by using kinetic maceration in ethanol. Antibacterial activity of CTEE against S. mutans was tested using disk diffusion agar and microdilution assays. Quorum sensing system employed Chromobacterium violaceum bacteria to produce violacein, and CTEE at various concentrations was tested for its antiquorum sensing activity to inhibit the violacein production. Our results demonstrated that CTEE at 1 mg/mL showed a significant inhibition >90% against S. mutans, indicating its MIC value. For the quorum sensing system, CTEE at the lowest concentration (0.25 mg/mL) significantly inhibit up to 68% of violacein produced by C. violaceum. These data indicate that CTEE may act as a natural oral functional food with antibacterial and antiquorum sensing activities for the prevention of dental caries.

Identification of New Potential Inhibitors of Quorum Sensing through a Specialized Multi-Level Computational Approach.

Biofilms are aggregates of microorganisms anchored to a surface and embedded in a self-produced matrix of extracellular polymeric substances and have been associated with 80% of all bacterial infections in humans. Because bacteria in biofilms are less amenable to antibiotic treatment, biofilms have been associated with developing antibiotic resistance, a problem that urges developing new therapeutic options and approaches. Interfering with quorum-sensing (QS), an important process of cell-to-cell communication by bacteria in biofilms is a promising strategy to inhibit biofilm formation and development. Here we describe and apply an in silico computational protocol for identifying novel potential inhibitors of quorum-sensing, using CviR—the quorum-sensing receptor from Chromobacterium violaceum—as a model target. This in silico approach combines protein-ligand docking (with 7 different docking programs/scoring functions), receptor-based virtual screening, molecular dynamic simulations, and free energy calculations. Particular emphasis was dedicated to optimizing the discrimination ability between active/inactive molecules in virtual screening tests using a target-specific training set. Overall, the optimized protocol was used to evaluate 66,461 molecules, including those on the ZINC/FDA-Approved database and to the Mu.Ta.Lig Virtual Chemotheca. Multiple promising compounds were identified, yielding good prospects for future experimental validation and for drug repurposing towards QS inhibition.

Disrupting quorum sensing alters social interactions in Chromobacterium violaceum

Quorum sensing (QS) is a communication system used by bacteria to coordinate a wide panel of biological functions in a cell density-dependent manner. The Gram-negative Chromobacterium violaceum has previously been shown to use an acyl-homoserine lactone (AHL)-based QS to regulate various behaviors, including the production of proteases, hydrogen cyanide, or antimicrobial compounds such as violacein. By using combined metabolomic and proteomic approaches, we demonstrated that QS modulates the production of antimicrobial and toxic compounds in C. violaceum ATCC 12472. We provided the first evidence of anisomycin antibiotic production by this strain as well as evidence of its regulation by QS and identified new AHLs produced by C. violaceum ATCC 12472. Furthermore, we demonstrated that targeting AHLs with lactonase leads to major QS disruption yielding significant molecular and phenotypic changes. These modifications resulted in drastic changes in social interactions between C. violaceum and a Gram-positive bacterium (Bacillus cereus), a yeast (Saccharomyces cerevisiae), immune cells (murine macrophages), and an animal model (planarian Schmidtea mediterranea). These results underscored that AHL-based QS plays a key role in the capacity of C. violaceum to interact with micro- and macroorganisms and that quorum quenching can affect microbial population dynamics beyond AHL-producing bacteria and Gram-negative bacteria.

Micro and nanoemulsions of Carissa spinarum fruit polyphenols, enhances anthocyanin stability and anti-quorum sensing activity: Comparison of degradation kinetics

The low stability of anthocyanins is a constraint in the food industry. The present work has been carried out to overcome this low stability by encapsulating fruit concentrate of underutilized plant Carissa spinarum (CS) with polyphenols in microemulsions (CSME) and nanoemulsions (CSNE). Increasing the amount of CS reduced the particle size from 1154 to 70–300 nm whereas addition of Tween 80 reduced it optimally to 5–25 nm. Degradation of anthocyanins in control and ME/NE proceeded with zero- and first-order reaction rates, respectively, at 28 °C (half-life 6, 25 and 40 days, respectively). The degradation kinetics of phenolics-flavonoids were also studied. CSNE exhibited higher anti-quorum sensing (QS) activity than CSME against Chromobacterium violaceum (73.7%); it inhibited biofilm formation by 70.1 and 64.4% in Pseudomonas aeruginosa, and Yersinia enterocolitica, respectively. This is the first report of using the more stable ME/NE to study anti-QS activity, an alternative to conventional antibiotics.