Inhibition Of Violacein Production Research Articles

Inhibition of biofilm and quorum sensing-regulated virulence factors in Pseudomonas aeruginosa by Cuphea carthagenensis (Jacq.) J. F. Macbr. Leaf extract: An in vitro study

Ethnopharmcological relevanceMicrobial biofilm formation, a quorum sensing (QS) regulated process, is one of the major causes of nosocomial and chronic infections, foodborne diseases, and associated deaths. Various approaches have been used to eradicate the menace of biofilm. Ethnomedicinal plants as potent antibiofilm agents are gaining a lot of interest in an era where the drug resistance is increasing and the availability of potent antibiotics is no longer promised. In this context, the methanol extract of Cuphea carthagenensis (CCMD), an ethno-medicinal and culinary herb, was evaluated as an antibiofilm and anti-QS agent against Pseudomonas aeruginosa.Aim of the study: The aim of the study is to evaluate the antibiofilm and anti-QS activity of an ethnomedicinal plant against a strong biofilm forming microorganism, P. aeruginosa. MethodsAntibiofilm activity of CCMD was demonstrated at different concentrations by Tissue Culture Plate, Test Tube method and other microscopic techniques. The effect of CCMD on QS and QS-related virulence factors viz. Pyocyanin, exopolymeric substance matrix (EPS), total protease, elastase, pyoverdin and swimming motility in P. aeruginosa were also evaluated. Antioxidant activity (DPPH & FRAP), total phenolic and flavonoid content were also checked. In order to determine the composition of the extract HPLC analysis was also performed. ResultsIn vitro study demonstrated a significant inhibition of biofilm formation (81.88 ± 2.57%) as well as production of QS-dependent virulence factors in P. aeruginosa. The extract also inhibited violacein production (83.31 ± 2.77%) in Chromobacterium violaceum which correlates with the reduction in QS-mediated virulence factors. The extract showed 64.79% ± 0.83% DPPH scavenging activity and reduction of ferricyanide complex (Fe3+) to the ferrous form (Fe2+) in DPPH and FRAP assay, respectively. Furthermore, the extract showed thermal stability and does not have any growth inhibitory effect on P. aeruginosa. The HPLC analysis demonstrated the presence of ellagic acid, ascorbic acid and hippuric acid in the extract. ConclusionThis work is the first to demonstrate that C. carthagenensis can attenuate biofilm formation and QS-mediated virulence factors of P. aeruginosa. Further investigation is required to use this ethnomedicinal plant (CCMD) as an important source of antibiofilm agents.

Ethanol Extract of Campsis grandiflora Flower and Its Organic Acid Components Have Inhibitory Effects on Autoinducer Type 1 Quorum Sensing.

Chinese herbs are a useful resource bank for natural drug development, and have attracted considerable attention to exploit quorum sensing inhibitors (QSIs). This study was designed to screen QSIs from raw Chinese herb materials. Of the 38 common herbs examined, the ethanol extract of Campsis grandiflora flower had the strongest QSI activity. The C. grandiflora flower ethanol extract (CFEE) was purified by HPD600, and the QSI activities were examined in further detail. CFEE inhibited violacein production of Chromobacterium violaceum 026 in a dose-dependent manner, and inhibit the swarming abilities of Escherichia coli K-12 and Pseudomonas aeruginosa PAO1. Furthermore, CFEE could inhibited biofilm formation and destroyed mature biofilms of E. coli K-12 and P. aeruginosa PAO1. The composition of CFEE was determined by UPLC-MS/MS to distinguish active QSI compounds, and 21 compounds were identified. In addition to gallic acid and caffeic acid, two organic acids, malic acid and succinic acid, were confirmed for the first time to have autoinducer type 1 QSI activities. Therefore, CFEE is a potential QSI that could be used as a novel antimicrobial agent and should be considered for medicinal development.

Pullulan Films Containing Rockrose Essential Oil for Potential Food Packaging Applications.

Active packaging is designed to control the development of decay- and disease-causing microorganisms and is emerging as a promising technology for extending shelf-life, maintaining food safety, reducing waste, and minimizing the risks for foodborne diseases. The goal of this work was to develop and characterize bioactive pullulan-based films, containing rockrose (Cistus ladanifer) essential oil. Among other abundant compounds (camphene, bornyl acetate and trans-pinocarveol), α-pinene was identified as the major compound of rockrose essential oil (39.25%). The essential oil presented stronger antibacterial activity against Gram-positive than against Gram-negative bacteria. The antioxidant results indicate the potential of the developed films to be used to package foods susceptible to oxidation and rancification, thus improving their shelf-life. Also, this study reflects the potential of rockrose essential oil, free or incorporated in pullulan, as a promising quorum sensing inhibitor, since it was able to interrupt intercellular communication, inhibiting violacein production. Electronic microscopy images showed the antibiofilm activity of the films with rockrose essential oil that were able to influence bacterial adhesion, which may be explained by the differences in the surface free energy of the films, as also determined.

Black cardamom essential oil prevents Escherichia coli O157:H7 and Salmonella Typhimurium JSG 1748 biofilm formation through inhibition of quorum sensing.

This study aimed to investigate the chemical composition, using GC-MS, and anti-biofilm potential of black cardamom essential oil (BCEO) against biofilms of Escherichia coli O157:H7 and Salmonella Typhimurium JSG 1748 through inhibition of bacterial quorum sensing. GC-MS quantification demonstrated that BCEO contains 1,8-cineole (44.24%), α-terpinyl acetate (12.25%), nerolidol (6.03%), and sabinene (5.96%) as the major bioactive compounds. Antioxidant assays for BCEO revealed the total phenolic and flavonoid mean values were 1325.03 ± 7.69mg GAE 100/g and 168.25 ± 5.26mg CE/g, respectively. In regards to antimicrobial potential, Candida albicans was the most sensitive species compared to Streptococcus mutans, Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, and Salmonella Typhimurium with the following zones of inhibition; 14.4 ± 0.52, 13.2 ± 0.42, 11.2 ± 0.28, 11.0 ± 0.52, 8.2 ± 0.24 and 6.6 ± 0.18mm in diameter, respectively. Biofilm inhibition by BCEO was concentration-dependent, when various concentrations of 0.03, 0.06, 0.12, 0.25 and 0.5% were applied, 33.67, 34.14, 38.66, 46.65 and 50.17% of Salmonella Typhimurium biofilm was inhibited, while 47.31, 54.15, 76.57, 83.36 and 84.63% of Escherichia coli biofilm formation was prevented. Chromobacterium violaceum ATCC 12,472 and its product violacein, was used as a microbial indicator for enhancement or inhibition of quorum sensing. Our data showed that 0.5% of BCEO inhibited violacein production without influencing the growth of Chromobacterium violaceum, while 1% of BCEO, caused 100% inhibtion of violacein production together with 30% inhibition of growth. This study shows that BCEO possesses promising antioxidant and antimicrobial potential, and found anti-biofilm activities linked to the quenching of the quorum sensing system of E. coli and S. Typhimurium.

Antimicrobial, anti-biofilm and anti-quorum sensing compounds from the bark of Ekebergia capensis

The main objective of this study was to isolate bioactive compounds from Ekebergia capensis for antimicrobial, anti-biofilm and anti-quorum sensing activity. The bark of E. capensis was extracted using hexane, ethyl acetate and methanol for 72 hours on maceration at room temperature. Antibacterial activity was evaluated against Staphylococcus aureus and antifungal activity against Candida albicans at 5 and 10 mg/mL concentrations. Anti-biofilm activity was tested using test tube and swarming motility test methods and anti-quorum sensing was evaluated using flask incubation test for violacein inhibition production methods. The results showed ethyl acetate extract of E. capensis and its two isolated compounds exhibited antimicrobial, anti-biofilm and anti-quorum sensing activity. Inhibition zones of antimicrobial activities of the extract against S. aureus and C. albicans ranged between 4-20 mm. 10 mg/mL extract showed inhibition of biofilm formation against S. aureus, anti-swarming activities against P. aeruginosa and S. aureus. Inhibition of violacein production against S. aureus, P. aeruginosa, and C. albicans were 72.6%, 67.7% and 69.3% respectively. The ethyl acetate extract, which showed antimicrobials, anti-biofilm, anti-quorum sensing activities, after silica gel column chromatography furnished two compounds namely; diisobutylphtalate and Sandropin B. The chemical structures of these compounds were done using spectroscopic methods including IR and NMR. The isolated compounds also showed strong antimicrobial, anti-biofilm, and anti-quorum sensing activity. This could be useful to manage infectious diseases caused by biofilm forming microorganisms.

Bioactive Properties of Syzygium cumini (L.) Skeels Pulp and Seed Phenolic Extracts.

The emergence of bacterial strains resistant to different antibiotics has prompted the search for new sources of antimicrobial compounds. Studies have shown that jambolan [Syzygium cumini (L.) Skeels], a tropical fruit from the Mirtaceae family, contains a great variety of phytochemical compounds with high antioxidant and antimicrobial activity. This study aimed to determine the centesimal composition and physicochemical characteristics of the pulp and seed of S. cumini (L.) Skeels, as well as the content of total phenolic compounds and the antioxidant, antibacterial, antibiofilm and anti-quorum sensing (QS) activities of the phenolic extracts obtained from the pulp and the seeds of this fruit. The in vitro antibacterial and anti-QS activities of active films incorporating phenolic extracts were also evaluated. Additionally, we performed molecular docking of phenolic compounds present in jambolan with the CviR QS regulator of Chromobacterium violaceum. The composition and physicochemical characteristics of the samples presented similar values to those found for the species. However, the seed phenolic extract had a higher content of phenolic compounds and antioxidant activity than the pulp. Both phenolic extracts presented antibacterial activity against Aeromonas hydrophila, C. violaceum, Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica serovar Typhimurium, Serratia marcescens, Listeria monocytogenes, and Staphylococcus aureus. The seed phenolic extract was particularly inhibitory against S. aureus. The pulp phenolic extract inhibited swarming motility and biofilm formation of A. hydrophila, E. coli, and S. marcescens in sub-MIC concentrations. The pulp and seed phenolic extracts inhibited violacein production in C. violaceum. Films incorporating both phenolic extracts inhibited the growth of bacteria, particularly Pseudomonas fluorescens, L. monocytogenes, and S. aureus, as well as QS in C. violaceum. Molecular docking showed that a variety of compounds found in pulp and seed extracts of jambolan, particularly chlorogenic acid and dihydroquercetin, potentially bind CviR protein and may interfere with QS. Our results indicate that pulp and seed of jambolan are good sources of antibacterial, antibiofilm, and anti-QS compounds that can be used in the development of natural preservatives and for application in antibacterial active films.

Thai herbal formulation ‘Ya-Pit-Samut-Noi’: Its antibacterial activities, effects on bacterial virulence factors and in vivo acute toxicity

Ethnopharmacological relevanceA Thai herbal formulation ‘Ya-Pit-Samut-Noi’ containing Nigella sativa (seed), Piper retrofractum (fruit), Punica granatum (pericarp), and Quercus infectoria (nutgall) has long been traditionally used to treat diarrhea or bloody mucous diarrhea. Scientific information is very important to support its therapeutic effects and traditional drug development. Aim of the studyThis study aimed to evaluate the antibacterial activities of Ya-Pit-Samut-Noi against diarrhea-causing bacteria and determine its effects on bacterial virulence factors and in vivo acute toxicity. Materials and methodsEthanol and water extracts of Ya-Pit-Samut-Noi and its plant components were prepared. The agar diffusion method was used for preliminary screening of antibacterial activity of the extracts against diarrhea-causing bacteria including Staphylococcus aureus, Vibrio cholerae, and Vibrio parahaemolyticus. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were assessed using broth microdilution method. The effects on bactericidal activity, bacterial cell wall, and cell membrane were examined by time-kill, lysis, and leakage assays, respectively. The effects on bacterial virulence factors including quorum-sensing system, biofilm production, and swarming motility were determined. Phytochemical screening was carried out to identify the group of chemical compounds present in the formulation extracts. Acute toxicity study was conducted by a single oral dose of 2000 mg/kg body weight in Wistar albino rats. ResultsEthanol and water extracts of Ya-Pit-Samut-Noi and Quercus infectoria demonstrated antibacterial efficacy against all bacterial strains as revealed by zones of inhibition ranging from 7.0 to 24.5 mm. The ethanol and water extracts of Ya-Pit-Samut-Noi and Quercus infectoria produced strong bacteriostatic activity against V. parahaemolyticus (n = 11) with an MIC range of 7.81–250 μg/ml. Only the ethanol extract of Ya-Pit-Samut-Noi produced MBC values less than or equal to 1000 μg/ml against all V. parahaemolyticus. Based on time-kill study, no surviving V. parahaemolyticus (ATCC 17802 and 5268) cells were detected within 6–12 h after treatment with the ethanol extract of Ya-Pit-Samut-Noi at MBC-4MBC concentrations. Vibrioparahaemolyticus ATCC 17802 cells treated with the ethanol extract of Ya-Pit-Samut-Noi demonstrated no lysis or leakage through the bacterial membrane was not observed. At low concentrations (0.125–0.25 μg/ml) the ethanol extract of Ya-Pit-Samut-Noi inhibited violacein production by Chromobacterium violaceum DMST 21761 without affecting the bacterial growth. The ethanol (31.25–62.5 μg/ml) and water (31.25–250 μg/ml) extracts of Ya-Pit-Samut-Noi inhibited biofilm production by S. aureus. The ethanol and water extracts of Ya-Pit-Samut-Noi at 1000 μg/ml reduced the swarming motility of Escherichia coli O157: H7 by 74.98% and 52.65%, respectively. Tannins and terpenoids were detected in both the ethanol and water extracts. Flavonoids were present only in the ethanol extract. Alkaloids and antraquinones were not noticed in either extract. In the acute toxicity study, there were no significant changes in hematological and biochemical parameters nor were adverse effects on mortality, general behaviors, body weight, or organ weights detected. ConclusionsThe scientific evidence from this study supported the therapeutic effects and safety of the traditional Thai herbal formulation ‘Ya-Pit-Samut-Noi’ which has been used as an alternative treatment for gastrointestinal infections in Thailand.

Green synthesis of silver nanoparticles using Carum copticum: Assessment of its quorum sensing and biofilm inhibitory potential against gram negative bacterial pathogens

Antimicrobial resistance among pathogenic bacteria has become a global threat to human health. Due to poor progress in development of new antimicrobial drugs, there is a need for the development of novel alternative strategies to combat the problem of multidrug resistance. Moreover, there is focus on ecofriendly approach for the synthesis nanoparticles having efficient medicinal properties including antivirulence properties to tackle the emergence of multi-drug resistance. Targeting quorum sensing controlled virulence factors and biofilms has come out to be a novel anti-infective drug target. The silver nanoparticles (Ag@CC-NPs) were synthesized from aqueous extract of Carum copticum and characterized using UV–vis absorption spectroscopy, fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Ag@CC-NPs were checked for its ability to inhibit quorum sensing-mediated virulence factors and biofilms against three test pathogens at sub-MIC values. There was ~75% inhibition of violacein production by Ag@CC-NPs against C. violaceum. The P. aeruginosa virulence factors such as pyocyanin production, pyoverdin production, exoprotease activity, elastase activity, swimming motility and rhamnolipid production were inhibited by 76.9, 49.0, 71.1, 53.3, 89.5, and 60.0% at sub-MIC. Moreover, virulence factors of S. marcescens viz. prodigiosin production, exoprotease activity, and swarming motility was reduced by 78.4, 67.8, and 90.7%. Ag@CC-NPs also exhibited broad-spectrum antibiofilm activity with 77.6, 86.3, and 75.1% inhibition of biofilms of P. aeruginosa, S. marcescens, and C. violaceum respectively. The biofilm formation on glass coverslip was reduced remarkably as evident from SEM and CLSM analysis. The findings revealed the in vitro efficacy of Ag@CC-NPs against bacterial pathogens and can be exploited in the development of alternative therapeutic agent in management of bacterial infections for topical application, mainly wound infection, or coating of surfaces to prevent bacterial adherence on medical devices.

Anti-quorum sensing and biofilm inhibitory activity of Apium graveolens L. oleoresin.

Apium graveolens L. (Apiaceae) is a dietary herb used as a spice, condiment and medicine. A. graveolens (Celery) has been studied for its antimicrobial property and for its application as flavours in food industry. The present study investigated the Apium graveolens oleoresin as an anti-quorum sensing and antibiofilm agent. The quorum sensing and biofilm inhibition study was carried out using biosensor strains Chromobacterium violaceum CV12472 and Pseudomonas aeruginosa PAO1. The MIC of celery oleoresin against C. violaceum CV12472 and P. aeruginosa PAO1 was 10 and 25% v/v, respectively. Inhibition of violacein and biofilm formation was tested at concentrations of oleoresins ranging from 1.56 and 50% v/v. The oleoresins showed a concentration dependent QS inhibitory activity and at sub-MIC of 6.25 and 12.5% v/v, the oleoresins significantly inhibited violacein production and biofilm formation (p < 0.05). Similarly, the celery oleoresin had significant QS modulatory effect on swimming, swarming and twitching motility in P. aeruginosa PAO1 at 12.5% v/v (p < 0.05). The major phytoconstituents present in celery oleoresin as analysed by GC-MS were eicosadiene, benzenemethanol and methyl ester which have not been previously reported. The findings suggest that celery has QS and biofilm inhibitory potential against gram negative pathogens and can find application as food intervention techniques.

Anti-quorum sensing and antimicrobial activities of South African medicinal plants against uropathogens

Urinary tract infections (UTIs) primarily affect women and have increasingly become a serious health problem globally. These infections are largely attributed to the quorum sensing (QS)-dependent ability of pathogens to form biofilms in the urinary tract. Microbial pathogenicity can be attenuated by disturbing the QS system of bacteria. The aim of the study was to document the antibacterial and anti-quorum sensing (AQS) potential of medicinal plants that are used as traditional medicine in South Africa to treat UTIs. Of the 70 plant extracts prepared from 14 medicinal plants using solvents of different polarities, only 6 displayed noteworthy activity (MIC < 1 mg/mL) against Pseudomonas aeruginosa, Staphylococcus aureus, Serratia marcescens, Escherichia coli and Proteus mirabilis. Extracts of Hydnora africana, Hypoxis hemerocallidea and Prunus africana displayed good antibacterial activity with MIC value of 0.5 mg/mL against S. aureus, but the methanol extracts of Rhoicissus tridentata and Cryptocarya latifolia were highly active against S. marcescens with MICs of 0.13 and 0.25 mg/mL, respectively. When plant extracts were screened for their ability to inhibit the QS-controlled violacein production by Chromobacterium violaceum, only two species (H. africana and C. latifolia) exhibited AQS activity in the qualitative agar well diffusion assay. However, eight extracts inhibited violacein production by 57–71% in the quantitative dilution assay. The ability of uropathogens to form biofilms upon exposure to the plant extracts was subsequently investigated using the crystal violet assay. It was found that the polar extracts of Cenchrus ciliaris and Eucomis autumnalis, Ranunculus multifidus, Vernonia adoensis, Cryptocarya latifolia, Hydnora africana, Rhoicissus tridentata and Hypoxis hemerocallidea, as well as non-polar extracts of Prunus africana and Hypoxis hemerocallidea were able to reduce initial cell attachment of S. aureus, P. mirabilis and S. marcescens by approximately 50%. However, the preformed biofilm was inhibited less than 30% by the extracts. The study revealed that several South African medicinal plants have antibacterial and AQS properties, validating their use in traditional medicines to treat UTIs to some degree, and indicating that they may be a suitable source of anti-pathogenic drugs to treat urinary infections.

Comparative Study of Inhibitory Potential of Dietary Phytochemicals Against Quorum Sensing Activity of and Biofilm Formation by Chromobacterium violaceum 12472, and Swimming and Swarming Behaviour of Pseudomonas aeruginosa PAO1.

SUMMARYQuorum sensing (QS) and biofilm formation are important mechanisms related to antibiotic resistance of many pathogens. Alternative treatments are needed to prevent recurrent or chronic infections caused by multi-resistant pathogens. Therefore, the aim of this study is to investigate and compare the inhibitory potential of the dietary phytochemicals: curcumin, quercetin, apigenin, pyrogallol, gallic acid and luteolin against QS of and biofilm formation by Chromobacterium violaceum ATCC 12472 and the swimming and swarming abilities of Pseudomonas aeruginosa PAO1. Anti-QS potential of the phytochemicals was evaluated qualitatively and quantitatively using C. violaceum via the disk diffusion assay based on violacein pigment inhibition at the subminimal inhibitory concentrations ranging from 46.87 to 750 µg/mL. The results of anti-QS and antibiofilm activities on C. violaceum demonstrated that all the phytochemicals except pyrogallol and gallic acid inhibited violacein production (from (11.0±0.1) to (88.2±0.1) %) in a concentration-dependent manner. In addition, the biofilm formation was also significantly inhibited (p<0.05) in the presence of all the phytochemicals ((1.38±0.08)–(84.2±0.2) %). In the present study, the results revealed that quercetin, curcumin, apigenin and luteolin could be promising QS and biofilm inhibitory agents against the C. violaceum 12472 biosensor system. Our findings also suggest that all the phytochemicals, especially curcumin, quercetin and pyrogallol, might be anti-pathogenic agents against P. aeruginosa PAO1 infections due to the ability to control QS. However, more comprehensive studies at the molecular level, explaining their anti-QS mechanisms, need to be conducted to confirm these results and identify the genes involved.

Anti-Quorum Sensing Activity of 12 Essential Oils on chromobacterium violaceum and Specific Action of cis-cis-p-Menthenolide from Corsican Mentha suaveolens ssp. Insularis.

Quorum sensing (QS) is a bacterial communication mechanism used to express various survival or virulence traits leading to enhanced resistance. Chromobacterium violaceum is a commonly used strain that highlights anti-QS action of bioactive substances. Here, we wanted to see if 12 selected essential oils (EO) could exert anti-QS activity. We measured the sublethal minimal QS inhibitory concentration (MQSIC) by assessing violacein production of C. violaceum along with bacterial growth. To confirm the QS disruption, we also proceed to surface bacterial observations using scanning electron microscopy (SEM). We showed that cis-cis-p-menthenolide extracted and isolated from a plant endemic to occidental Mediterranean Sea islands, Mentha suaveolens ssp. insularis, acts as an inhibitor of violacein production and biofilm formation. Measured MQSIC was much lower than the minimal inhibitory concentration (MIC): 0.10 mg·mL−1 vs. 3.00 mg·mL−1. Moreover, disturbance of QS-related traits was confirmed by the degradation of C. violaceum biofilm matrix. There is a clear structure–activity relationship between cis-cis-p-menthenolide and anti-QS activity. Indeed, its isomer molecule (mintlactone) exerts a poor anti-QS action. These results indicate that inhibition of violacein production and biofilm formation by cis-cis-p-menthenolide might be related to a disruption in the QS mechanism.

The Effects of Chinese Herbal Medicines on the Quorum Sensing-Regulated Virulence in Pseudomonas aeruginosa PAO1.

The quorum sensing (QS) system has been used by many opportunistic pathogenic bacteria to coordinate their virulence determinants in relation to cell-population density. As antibiotic-resistant bacteria are on the rise, interference with QS has been regarded as a novel way to control bacterial infections. As such, many plant-based natural products have been widely explored for their therapeutic roles. These natural products may contain anti-QS compounds that could block QS signals generation or transmission to combat QS pathogens. In this study, we report the anti-QS activities of four different Chinese herbal plant extracts: Poria cum Radix pini, Angelica dahurica, Rhizoma cibotii and Schizonepeta tenuifolia, on Pseudomonas aeruginosa PAO1. All the plants extracted using hexane, chloroform and methanol were tested and found to impair swarming motility and pyocyanin production in P. aeruginosa PAO1, particularly by Poria cum Radix pini. In addition, all the plant extracts also inhibited violacein production in C. violaceum CV026 up to 50% while bioluminescence activities were reduced in lux-based E. coli biosensors, pSB401 and pSB1075, up to about 57%. These anti-QS properties of the four medicinal plants are the first documentation that demonstrates a potential approach to attenuate pathogens’ virulence determinants.

Headspace analysis, antimicrobial and anti-quorum sensing activities of seven selected African Commiphora species

The genus Commiphora has been widely used in traditional medicinal systems to treat pain, skin infections, diarrhoea and periodontal diseases. Oleo-gum resins of seven Commiphora species (C. myrrha, C. confusa, C. baluensis, C. pseudopaolii, C. guidotti, C. holtziana and C. kataf) were collected from Kenya and Ethiopia. Headspace analysis was determined using GCxGC–ToF–MS. In vitro evaluation of antimicrobial activity of acetone extracts were evaluated against a range of micro-organisms using microdilution assay. In addition, anti-quorum sensing activity of the tested Commiphora species was assessed qualitatively and quantitatively using Chromobacterium violaceum strain. α-Pinene (5.6–8.1%), β-caryophyllene (nd-16.5%), cis-β-ocimene (nd-7.4%), α-santalene (nd-40.8%), α-bourbonene (6.9–37.4%), β-elemene (nd-26.2%), curzerene (4.5–8.2%), p-cymene (1.8–54.5%), curzerenone (1.6–25.5%), (Z)-α-bisabolene (nd-10.4%), β-selinene (nd-9.8%), germacrene D (nd-8.4%), cresol methyl ether (nd-8.5%), 7-epi-sesquithujene (nd-8.8%), dehydro-aromadendrene (nd-36.9%), δ-elemene (nd-31.1%) and spathulenol (0.2–10.4%) were the major volatile constituents. Principal component analysis showed three clusters corresponding to three chemotypes within the dataset. The extracts exhibited good to moderate antibacterial activity against a panel of selected micro-organisms (MIC values: 0.06–8.0mg/mL). Commiphora holtziana displayed the best activity against Candida tropicalis (0.06–4.0mg/mL), while C. holtziana and C. myrrha exhibited the best anti-quorum sensing activity by inhibiting violacein production.

Quorum sensing inhibitors from marine bacteria Oceanobacillus sp. XC22919

In this study, three active compounds isolated from Oceanobacillus sp. XC22919 were identified as 2-methyl-N-(2′-phenylethyl) butyramide (1), 3-methyl-N-(2′-phenylethyl)-butyramide (2) and benzyl benzoate (3), and were first reported to exhibit the apparent quorum sensing inhibitory activities against C. violaceum 026 and P. aeruginosa. Compounds 1–3 inhibited violacein production of C. violaceum 026 by 10.5–55.7, 11.2–55.7, and 27.2%–95.7%, respectively, and inhibited pyocyanin production of P. aeruginosa by 1.7–50.8, 39.1–90.7, and 57.2%–98.7%, respectively. The azocasein-degrading proteolytic rates of P. aeruginosa were observed by 13.4–31.5, 13.4–28.8, and 11.3%–21.1%, respectively. With respect to elastase, the range of inhibition of activity of compounds 1–3 was 2.1–30.3, 4.2–18.2, and 8.9%–15.7%, respectively. Compounds 1 and 3 also showed a concentration-dependent attenuation in biofilm formation, with the maximum of 50.6% inhibition, and 37.7% inhibition at 100 μg/mL, respectively.

Hexanal as a QS inhibitor of extracellular enzyme activity of Erwinia carotovora and Pseudomonas fluorescens and its application in vegetables

To prevent the postharvest disease of Chinese cabbage and lettuce, hexanal was used as a control measure to inhibit N-acyl homoserine lactone (AHL) production and extracellular enzymes regulated by quorum-sensing (QS) in their main spoilage strains of Erwinia carotovora and Pseudomonas fluorescens. Firstly, the QS inhibition of hexanal was verified by significantly inhibiting violacein production (p < 0.05) in Chromobacterium violaceum CV026 at sub-MICs. β-Galactosidase activities which reflected AHL production, were significantly inhibited by hexanal, its inhibitory effect was concentration-dependent under minimal inhibitory concentration (MIC) (p < 0.05). The detected extracellular enzymes activities decreased with the increase of hexanal concentration (p < 0.05), including cellulase, xylanase, pectate lyase, polygalacturonase, and protease. Chinese cabbage soft rot and lettuce leaf scorch could be significantly inhibited by hexanal (p < 0.05) without any phytotoxicity effect, the 1/2 MIC of hexanal showed the best inhibitory effect. And all the above effects showed a dose-dependent. A novel preservation technique in reducing the loss of vegetables due to spoilage based on the QS inhibitor was developed.

Quorum sensing inhibitory potential and in silico molecular docking of flavonoids and novel terpenoids from Senegalia nigrescens

Ethnopharmacological relevanceSenegalia nigrescens is used in traditional medicine for the treatment of dysentery and convulsions. Aims of the studyThis study was aimed at identifying bioactive compounds from S. nigrescens and carrying out in vitro and in silico anti-quorum sensing studies on the compounds. Materials and methodsExtracts of S. nigrescens were chromatographed repeatedly. The isolated compounds were characterised using NMR spectroscopy and mass spectrometry. The anti-quorum sensing potential of S. nigrescens crude extracts and selected phytochemicals was quantified using Chromobacterium violaceum quorum sensing-controlled violacein inhibition assays. Qualitative modulation of quorum sensing activity and signal synthesis was investigated using agar diffusion double ring assays and C. violaceum. Molecular docking was conducted to explore the binding conformations of ent-kaurene diterpenes and flavonoids into the binding sites of quorum sensing regulator proteins, CviR and CviR′. ResultsPhytochemical investigation of S. nigrescens resulted in the isolation of a new ent-kaurene diterpenoid (ent-kaur-15-en-18,20-diol) alongside ent-kaur-15-en-18-ol, being isolated for the first time from a plant species. Other compounds isolated included 30-hydroxylup-20(29)-en-3β-ol, 3β-hydroxy-20(29)-en-lupan-30-al, lupeol, stigmasterol, a long chain alcohol (tetracosan-1-ol) and three flavonoids (melanoxetin, quercetin and quercetin-3-O-methyl ether). Structures of isolated compounds were elucidated using different spectroscopic techniques including 1D and 2D NMR. Inhibition of violacein production was concentration-dependent, with 56.52% inhibition being obtained with 200 µg of quercetin-3-O-methyl ether, while 53.38% inhibition was obtained with 600 µg of quercetin. Agar diffusion double ring assays indicated CviI synthase/CviR receptor modulation by S. nigrescens phytochemicals, suggesting that quorum signal synthesis was down-regulated and/or targeting binding of signal to the receptor. The computed binding energy data suggested that the flavonoids had a stronger tendency to inhibit both CviR and CviR′ with varying binding affinities. ConclusionS. nigrescens crude extracts together with the novel ent-kaurenoids and flavonoids demonstrated potential anti-quorum sensing activity. S. nigrescens may thus represent a source of anti-quorum sensing therapeutic candidates for the control of existing and emerging infectious diseases.

Antioxidant, Antiquorum-Sensing and Antibiofilm Activities of &amp;lt;i&amp;gt;Balanites aegyptiaca&amp;lt;/i&amp;gt; (L.) Del. (Balanitaceae) and &amp;lt;i&amp;gt;Terminalia macroptera&amp;lt;/i&amp;gt; Guill. and Perr. (Combretaceae)

Pseudomonas aeruginosa like many pathogen bacteria produces various virulence factors and form biofilm that contribute to its pathogenicity and the growing resistance to antibiotics. The production of virulence factors in those multi-resistant bacteria is controlled by a mechanism of regulation termed quorum sensing. Interfering with this mechanism of bacterial communication constitute a strategy to attenuate bacterial pathogenicity. Our study aimed to assess the ability of Balanites aegyptiaca and Terminalia macroptera to interfere with the system of QS through inhibition of QS-controlled factors production and the formation of biofilm. Methanol extracts of galls, stem bark of B. aegyptiaca, and stem bark of T. macroptera were screened for anti-QS activity using Chromobacterium violaceum CV026 and P. aeruginosa PAO1. At a sub-inhibitory concentration of 100 µg/mL, galls and stem bark of B. aegyptiaca quenched the QS system by inhibiting violacein production on C. violaceum CV026 and pyocyanin production on P. aeruginosa PAO1. The bark of T. macroptera reduced significantly the production of violacein, pyocyanin and the formation of biofilm. Moreover, antioxidant activity of phenolic compounds contributes to reduce the oxidative stress induced by pyocyanin. Thus, T. macroptera is a potential candidate for the identification and isolation of news effective anti-QS compounds. This study introduces a possible validation for traditional use of B. aegyptiaca and T. macroptera and constitutes a new therapeutic approach for the treatment of infections caused by bacteria resistant to antibiotics.

In silico identification of albendazole as a quorum sensing inhibitor and its in vitro verification using CviR and LasB receptors based assay systems

Introduction:Quorum sensing inhibition (QSI) is one of the vital tools to overcome emerging virulence of pathogenic bacteria which aims at curbing bacterial resistance. Targeting QS (quorum sensing) as chemotherapy is less likely to generate resistance among pathogens as it targets only the adaptation and not the survival mechanism of the pathogen. Several QS inhibitors were developed in the recent past but none of them managed to have clinical application due to known toxic effects for human consumption. A rapid development of QS inhibitor drugs could be achieved by verification of the QSI activity of drugs which are already in clinical use with known pharmacology. Recently, a known FDA approved clinical drug niclosamide belonging to an anthelmintic class is found to exhibit QSI activity. Methods: We have focused our study on Albendazole, another FDA approved clinical drug belonging to the same class for its potential to act as QSI. The structure-based molecular docking is used for finding putative interactions made by this drug with the CviR and LasB receptor protein of Chromobacterium violaceum and Pseudomonas aeruginosa , respectively. Further, the in vitro activity of this drug has been evaluated by employing CviR and LasB receptor-based bioassay. The efficacy of this drug alone and in combination with antibiotic Tobramycin to inhibit P. aeruginosa based biofilms was also analyzed by developing the biofilms on chambered glass slides and performing anti-biofilm assay. Results: Further, this drug found to inhibit purple pigment violacein production in C. violaceum , which is under the control of C6-AHL-CviR mediated QS in this human pathogen. The in vivo bioassays results suggested that albendazole has great potential to act as a QS inhibitor as found inhibiting violacein production in C. violaceum and biofilm formation in P. aeruginosa , respectively. Conclusion: It is that structure-based molecular docking guided bioassay evaluation is an efficient tool for finding the new therapeutic use of old drugs which could have more chances to come easily in clinical application for their newly identified therapeutic uses.

Antiquorum Sensing Activity of Seed Oils from Oleaginous Plants and Protective Effect During Challenge with Chromobacterium violaceum.

Seed oils from oleaginous plants are rich in fatty acids (FAs) that play important roles in the health of the consumers. Recent studies indicate that FA also can play an important role in communication and regulation of virulence in bacteria. Nevertheless, evidence demonstrating protection against bacterial infections mediated by their quorum sensing inhibition (QSI) activity is scarce. In this study, sunflower, chia, and amaranth oils, were assayed for their QSI capacity by inhibiting violacein production and alkaline exoprotease activity of Chromobacterium violaceum. In vitro assays revealed that the oils exhibited QSI activities, whereas in vivo they delayed death of mice inoculated intraperitoneally with the bacterium. Gas chromatography coupled with mass spectrometry analysis of the oils indicated the presence of saturated FA (SAFA) and unsaturated FA as main components. Through a structure-activity relationship study of free FAs, bactericidal effect was identified mainly for polyunsaturated FAs, whereas QSI activity was restricted to SAFA of chains 12-18 carbon atoms in length. These data correlate with a possible interaction suggested by molecular docking analysis of lauric, myristic, and stearic acids with the CviR protein. Our study highlights the antiquorum sensing potential of SAFA, which may be future antivirulence therapeutic agents for the treatment of bacterial infections.