Anti-quorum sensing activity of some medicinal plants

Background: Quorum sensing is the key regulator of virulence factors of Pseudomonas aeruginosa such as biofilm formation, motility, productions of proteases, hemolysin, pyocyanin, and toxins. The aim of this study was to explore the effect of the extracts from some medicinal plants on quorum sensing and related virulence factors of P. aeruginosa.Material and Methods: Quorum sensing inhibitory (OSI) effect of the alcohol extracts of 20 medicinal plants was evaluated by Chromobacterium violaceum reporter using agar cup diffusion method. The efficient QSI extracts were tested for their activity against biofilm synthesis, motility, and synthesis of pyocyanin from P. aeruginosa PA14Results: The extracts of Citrus sinensis, Laurus nobilis, Elettaria cardamomum, Allium cepa, and Coriandrum sativum exhibited potent quorum quenching effect. On the other hand, Psidium guajava and Mentha longifolia extracts showed lower QSI activity. These extracts exhibited significant elimination of pyocyanin formation and biofilm development of Pseudomonas aeruginosa PA14. In addition, they significantly inhibited twitching and swimming motilities of P. aeruginosa PA14.Conclusion: This study illustrated, for the first time, the importance of C. sinensis, L. nobilis, E. cardamomum, A. cepa, and C. sativum as quorum sensing inhibitors and virulence suppressors of P. aeruginosa. Thus, these plants could provide a natural source for the elimination of Pseudomonas pathogenesis.Keywords: Quorum sensing inhibitory activity, P. aeruginosa, Chromobacterium violaceum, virulence factor

Quorum sensing inhibitory activity of sub-inhibitory concentrations of β-lactams

Introduction: The virulence factors of Pseudomonas aeruginosa are under the control of quorum sensing (QS) signals. Hence, interference with QS prevents its pathogenesis.Objective: The aim of the present research is to assess the influence of some β-lactam antibiotics on cell communication and the release of different virulence factors.Methods: The minimal inhibitory concentrations of ceftazidime, cefepime and imipenem were evaluated by microbroth dilution method. The effect of sub-inhibitory concentration of the tested antibiotics on QS signals was investigated using reporter strain assay. In addition, different virulence factors (elastase, protease, pyocyanin and hemolysin) were estimated in the presence of their sub-inhibitory concentrations.Results: Low concentrations of ceftazidime, cefepime and imipenem caused significant elimination of the QS signals 3OHC12- HSL and C4-HSL up to 1/20 MIC. Furthermore, low concentrations of the tested antimicrobials suppressed virulence factors elastase and hemolysin. Moreover, 1/20 of their MICs reduced elastase, protease, pyocyanin and hemolysin.Conclusion: Utilization of β-lactam antibiotics at low concentrations could be an effective approach for prevention and treatment of P. aeruginosa infection.Keywords: Quorum sensing inhibition, β-lactams, Pseudomonas aeruginos

Quorum sensing inhibitory activity of sub-inhibitory concentrations of \u3b2-lactams

Introduction: The virulence factors of Pseudomonas aeruginosa are under the control of quorum sensing (QS) signals. Hence, interference with QS prevents its pathogenesis. Objective: The aim of the present research is to assess the influence of some \u3b2-lactam antibiotics on cell communication and the release of different virulence factors. Methods: The minimal inhibitory concentrations of ceftazidime, cefepime and imipenem were evaluated by microbroth dilution method. The effect of sub-inhibitory concentration of the tested antibiotics on QS signals was investigated using reporter strain assay. In addition, different virulence factors (elastase, protease, pyocyanin and hemolysin) were estimated in the presence of their sub-inhibitory concentrations. Results: Low concentrations of ceftazidime, cefepime and imipenem caused significant elimination of the QS signals 3OHC12-HSL and C4-HSL up to 1/20 MIC. Furthermore, low concentrations of the tested antimicrobials suppressed virulence factors elastase and hemolysin. Moreover, 1/20 of their MICs reduced elastase, protease, pyocyanin and hemolysin. Conclusion: Utilization of \u3b2-lactam antibiotics at low concentrations could be an effective approach for prevention and treatment of P. aeruginosa infection

Antibiofilm activity of biosynthesized silver and copper nanoparticles using Streptomyces S29

Abstract Microbial resistance and biofilm formation have been considered as the main problems associated with microbial resistance. Several antimicrobial agents cannot penetrate biofilm layers and cannot eradicate microbial infection. Therefore, the aim of this study is the biological synthesis of silver and copper nanoparticles to assess their activities on bacterial attachment and on the viability of dormant cells within the biofilm matrix. Ag-NPs and Cu-NPs were biosynthesized using Streptomyces isolate S29. The biologically synthesized Ag-NPs and Cu-NPs exhibited brown and blue colors and were detected by UV/Vis spectrophotometry at 476 and 594 nm, respectively. The Ag-NPs showed an average size of 10–20 nm as indicated by TEM, and 25–35 nm for Cu-NPs. Both Ag-NPs and Cu-NPs were monodispersed with a polydispersity index of 0.1–0.546 and zeta potential were − 29.7, and − 33.7 mv, respectively. The biologically synthesized Ag-NPs and Cu-NPs significantly eliminated bacterial attachment and decreased the viable cells in the biofilm matrix as detected by using crystal violet and tri-phenyl tetrazolium chloride assays. Furthermore, Ag-NPs and Cu-NPs significantly eradicated mature biofilms developed by various Gram-negative pathogens, including A. baumannii, K. pneumoniae and P. aeruginosa standard strains and clinical isolates. Data were also confirmed at the molecular level with prominent elimination of biofilm gene expression carO, bssS and pelA in A. baumannii, K. pneumoniae and P. aeruginosa, respectively compared to untreated cells under the same conditions. As indicated, Ag-NPs and Cu-NPs could be used as adjuvant therapy in eradication of antibiotic resistance and biofilm matrix associated with Gram-negative bacterial infection. Graphical Abstrac

Inhibition of Quorum Sensing and Virulence Factors of Pseudomonas aeruginosa by Biologically Synthesized Gold and Selenium Nanoparticles

The development of microbial resistance requires a novel approach to control microbial infection. This study implies the microbial synthesis of nanometals and assessment of their antivirulent activity against Pseudomonas aeruginosa. Streptomyces isolate S91 was isolated from soil with substantial ability for growth at high salts concentrations. The cell-free supernatant of S91was utilized for the synthesis of Au-NPs and Se-NPs. The 16S rRNA sequence analysis of Streptomyces S91 revealed that S91 had a high similarity (98.82%) to Streptomyces olivaceous. The biosynthesized Au-NPs and Se-NPs were characterized using a UV-Vis spectrophotometer, dynamic light scattering, transmission electron microscopy, energy dispersive X-ray diffraction and Fourier-transform infrared spectroscopy. The quorum sensing inhibitory (QSI) potential of Au-NPs and Se-NPs and the antivirulence activity was examined against P. aeruginosa. The QSI potential was confirmed using RT-PCR. The synthesized Au-NPs and Se-NPs were monodispersed spherical shapes with particle size of 12.2 and 67.98 nm, respectively. Au-NPs and Se-NPs eliminated QS in P. aeruginosa at a concentration range of 2.3–18.5 µg/mL for Au-NPs and 2.3–592 µg/mL for Se-NPs. In addition, Au-NPs and Se-NPs significantly inhibited QS-related virulence factors, such as pyocyanin, protease and, elastase in P. aeruginosa. At the molecular level, Au-NPs and Se-NPs significantly suppressed the relative expression of QS genes and toxins. Hence, the biosynthesized Au-NPS and Se-NPS could be substantial inhibitors of QS and virulence traits of P. aeruginosa

Synthesis of silver nanoparticles with antimicrobial and anti-adherence activities against multidrug-resistant isolates from Acinetobacter baumannii

أهداف البحث: يمثل انتشار الجراثيم المقاومة للعقاقير المتعددة تهديدا صحيا كبيرا. وتعتبر جسيمات الفضة النانوية فئة حديثة من العوامل المضادة للجراثيم. وتهدف هذه الدراسة إلى التحضير الميكروبي لجسيمات الفضة النانوية، وتقييم أنشطتها المضادة للجراثيم والأغشية البكتيرية السطحية. طرق البحث: تم تصنيع جسيمات الفضة النانوية باستخدام محلول نمو “الاسينيتوباكتر بوماني” الخالي من الخلايا. وتم تمييز جسيمات الفضة النانوية عن طريق تحليل حجم الجسيمات وانتقال المجهر الإلكتروني، وتم تقييم الأنشطة المضادة للجراثيم، والأغشية البكتيرية السطحية لجسيمات الفضة النانوية المصنعة. النتائج: رصدت جسيمات الفضة النانوية أولا عن طريق تحويل اللون الأصفر الشاحب للمحلول الطافي الخالي من البكتيريا إلى اللون البني الداكن. وكان لجسيمات الفضة النانوية شكل كروي موحد وتراوح حجم الجسيمات من ٣.٧ إلى ١٦٨ نانومتر، وزيتا المحتملة- ١١.٧ مل فولت. كانت جسيمات الفضة النانوية المحضرة من الاسينيتوباكتر فعالة ضد البكتيريا المقاومة للأدوية المتعددة مثل البكتيريا القولونية، والزائفة الزنجارية، والكلبسيلة الرئوية والحد الأدنى من التركيزات المثبطة للنمو٣.١، ١.٥٦، ٣.١ ميكروغرام/ مل على التوالي. وعلاوة على ذلك، قضت جسيمات الفضة النانوية المحضرة من الاسينيتوباكتر بشكل كبير على أنشطة الربط للبكتيريا القولونية، والزائفة الزنجارية والكلبسيلة الرئوية بنسبة ٦٦.٦٪، ٨٦.٥ ٪ و٧٥٪ على التوالي. الاستنتاجات: منعت جسيمات الفضة النانوية، المحضرة من الاسينيتوباكتر بومانى نمو الجراثيم وقضت على تجمع الأغشية البكتيرية السطحية بواسطة عزلات المجرى البولي المقاومة للعقاقير المتعددة. وتشير هذه النتائج إلى إمكانية استخدام جسيمات الفضة النانوية كمضاد للجراثيم والأغشية البكتيرية السطحية ضد العدوى الناجمة عن العزلات المقاومة

Imipenem/cilastatin encapsulated polymeric nanoparticles for destroying carbapenem-resistant bacterial isolates

Abstract Background Carbapenem-resistance is an extremely growing medical threat in antibacterial therapy as the incurable resistant strains easily develop a multi-resistance action to other potent antimicrobial agents. Nonetheless, the protective delivery of current antibiotics using nano-carriers opens a tremendous approach in the antimicrobial therapy, allowing the nano-formulated antibiotics to beat these health threat pathogens. Herein, we encapsulated imipenem into biodegradable polymeric nanoparticles to destroy the imipenem-resistant bacteria and overcome the microbial adhesion and dissemination. Imipenem loaded poly Ɛ-caprolactone (PCL) and polylactide-co-glycolide (PLGA) nanocapsules were formulated using double emulsion evaporation method. The obtained nanocapsules were characterized for mean particle diameter, morphology, loading efficiency, and in vitro release. The in vitro antimicrobial and anti adhesion activities were evaluated against selected imipenem-resistant Klebsiella pneumoniae and Pseudomonas aeruginosa clinical isolates. Results The obtained results reveal that imipenem loaded PCL nano-formulation enhances the microbial susceptibility and antimicrobial activity of imipenem. The imipenem loaded PCL nanoparticles caused faster microbial killing within 2–3 h compared to the imipenem loaded PLGA and free drug. Successfully, PCL nanocapsules were able to protect imipenem from enzymatic degradation by resistant isolates and prevent the emergence of the resistant colonies, as it lowered the mutation prevention concentration of free imipenem by twofolds. Moreover, the imipenem loaded PCL eliminated bacterial attachment and the biofilm assembly of P. aeruginosa and K. pneumoniae planktonic bacteria by 74 and 78.4%, respectively. Conclusions These promising results indicate that polymeric nanoparticles recover the efficacy of imipenem and can be considered as a new paradigm shift against multidrug-resistant isolates in treating severe bacterial infections

Ciprofloxacin- and levofloxacin-loaded nanoparticles efficiently suppressed fluoroquinolone resistance and biofilm formation in Acinetobacter baumannii

Abstract The spread of fluoroquinolone (FQ) resistance in Acinetobacter baumannii represents a critical health threat. This study aims to overcome FQ resistance in A. baumannii via the formulation of polymeric nanoFQs. Herein, 80 A. baumannii isolates were obtained from diverse clinical sources. All A. baumannii isolates showed high resistance to most of the investigated antimicrobials, including ciprofloxacin (CIP) and levofloxacin (LEV) (97.5%). FQ resistance-determining regions of the gyrA and parC genes were the most predominant resistant mechanism, harbored by 69 (86.3%) and 75 (93.8%) of the isolates, respectively. Additionally, plasmid-mediated quinolone resistance genes aac(6′)-Ib and qnrS were detected in 61 (76.3%) and 2 (2.5%) of the 80 isolates, respectively. The CIP- and LEV-loaded poly ε-caprolactone (PCL) nanoparticles, FCIP and FLEV, respectively, showed a 1.5–6- and 6–12-fold decrease in the MIC, respectively, against the tested isolates. Interestingly, the time kill assay demonstrated that MICs of FCIP and FLEV completely killed A. baumannii isolates after 5–6 h of treatment. Furthermore, FCIP and FLEV were found to be efficient in overcoming the FQ resistance mediated by the efflux pumps in A. baumannii isolates as revealed by decreasing the MIC four-fold lower than that of free CIP and LEV, respectively. Moreover, FCIP and FLEV at 1/2 and 1/4 MIC significantly decreased biofilm formation by 47–93% and 69–91%, respectively. These findings suggest that polymeric nanoparticles can restore the effectiveness of FQs and represent a paradigm shift in the fight against A. baumannii isolates

Quorum sensing inhibiting activity of Streptomyces coelicoflavus isolated from soil

Quorum sensing (QS) systems communicate bacterial population and stimulate microbial pathogenesis through signaling molecules. Inhibition of QS signals potentially suppresses microbial infections. Antimicrobial properties of Streptomyces have been extensively studied, however, less is known about quorum sensing inhibitory (QSI) activities of Streptomyces. This study explored the QSI potential of Streptomyces isolated from soil. Sixty-five bacterial isolates were purified from soil samples with morphological characteristics of Streptomyces. The three isolates: S6, S12, and S17, exhibited QSI effect by screening with the reporter, Chromobacterium violaceum. Isolate S17 was identified as Streptomyces coelicoflavus by sequencing of the hypervariable regions (V1-V6) of 16S rRNA and was assigned gene bank number KJ855087. The QSI effect of the cell-free supernatant of isolate S17 was not abolished by proteinase K indicating the non-enzymatic activity of QSI components of S17. Three major compounds were isolated and identified, using spectroscopic techniques (1D, 2D NMR and Mass spectrometry), as behenic acid (docosanoic acid), borrelidin and 1H-pyrrole-2-carboxylic acid. 1H-pyrrole-2-carboxylic acid inhibited QS and related virulence factors of Pseudomonas aeruginosa PAO1 including; elastase, protease and pyocyanin without affecting Pseudomonas viability. At the molecular level, 1H-pyrrole-2-carboxylic acid suppressed the expression of QS genes (lasI, lasR, lasA, lasB, rhlI, rhlR, pqsA and pqsR). Moreover, QSI activity of S17 was assessed under different growth conditions and ISP2 medium supplemented with glucose 0.4% w/v and adjusted at pH 7, showed the highest QSI action. In conclusion, 1H-pyrrole-2-carboxylic acid, one of the major metabolites of Streptomyces isolate S17, inhibited QS and virulence determinants of P. aeruginosa PAO1. The findings of the study open the scope to exploit the in vivo efficacy of this active molecule as anti-pathogenic and anti-virulence of P. aeruginosa

Immunization with outer membrane proteins (OprF and OprI) and flagellin B protects mice from pulmonary infection with mucoid and nonmucoid Pseudomonas aeruginosa

Background: Pseudomonas aeruginosa is a Gram-negative opportunistic bacterium, which considered as a common cause of nosocomial infection and life-threatening complications in immunocompromized and cystic fibrosis patients. Here, we evaluate the protective effect of recombinant vaccines composed of outer membrane proteins OprF and OprI alone or in combination with flagellin B against mucoid and nonmucoid pseudomonas infection. Methods: BALB/C mice were immunized subcutaneous using OprF and OprI with or without flagellin B and antibody titers were determined. Serum bactericidal and opsonophagocytosis activities of immunized and control sera were estimated against mucoid and nonmucoid pseudomonas strains. Lung tissue sections from immunized and nonimmunized mice were analyzed and the levels of peripheral neutrophils infiltration into the lung and tissue inflammation were scored. Results: Subcutaneous immunization using OprF and OprI with or without flagellin B elicited higher antibody titers against OprF, OprI, and flagellin B. The produced antibodies successfully opsonized both mucoid and nonmucoid strains with subsequent activation of the terminal pathway of complement that enhances killing of nonmucoid strains via complement-mediated lysis. Furthermore, opsonized mucoid and nonmucoid strains showed enhanced opsonophagocytosis via human peripheral neutrophils, a mechanism that kills P. aeruginosa when complement mediated lysis is not effective especially with mucoid strains. Immunized mice also showed a significant prolonged survival time, lower bacteremia, and reduced lung damage when compared with control nonimmunized mice. Conclusion: Our data showed that mice immunized with OprF/OprI or OprF/OprI and flagellin B are significantly protected from infection caused by mucoid and nonmucoid strains of P. aeruginosa