ABSTRACT
Swarming motility is a typical synergistic motion, in which bacteria use flagella and Type Ⅳ Pili together to move collectively on semi-solid surfaces. Swarming motility is a hot topic of research in the field of microbiology because of its close relationship with biofilm formation, fruiting bodies formation, pathogen invasion and microbial dispersal and symbiosis. A large number of studies have been conducted on bacterial swarming motility, including changes in the expression of key proteins, changes in chemical communications between bacteria as well as mechanical changes. The expression of flagellin and the level of intracellular c-di-GMP complicatedly regulates the collective behavior of bacteria in colonies, which consequently impacts the swarming motility. The unique physical properties of swarmer cells are conducive to the expansion of the whole colony. Factors such as nutrient and water content in the surrounding growth environment of bacteria also affect the ability of bacteria to swarm to different degrees. It is challenging to construct a universal model of swarming motility based on the molecular mechanisms of swarming in the future.
Subject(s)
Bacteria , Flagellin , Symbiosis , WaterABSTRACT
BACKGROUND AND OBJECTIVE@#Many opportunistic and nosocomial pathogens like Pseudomonas aeruginosa are very reliant on a bacterium-to-bacterium communication system called quorum sensing (QS). Without the aforementioned process, gene expressions associated with virulence factors will not be produced. In this study, the sub-inhibitory concentrations (sub-MICs) of methanolic leaf extract and obtained fractions from Averrhoa bilimbi (kamias) were screened for ability to inhibit quorum sensing-controlled phenotypes of P. aeruginosa ATCC 27853.@*METHODOLOGY@#A. bilimbi crude extract was fractionated through liquid-liquid extraction, producing four (4) fractions: hexane fraction, dichloromethane (DCM) fraction, ethyl acetate (EtOAc) fraction, and water (H2O) fraction. Among the sub-MICs obtained from resazurin-based fluorimetric microtiter assay, only 50 μg/mL was utilized in evaluating the anti-QS properties of crude extract and fr@*RESULTS@# In the swarming motility assay, hexane fraction (9.39 mm ± 0.67) and DCM fraction (10.82 mm ± 0.95) displayed restriction in the treated P. aeruginosa ATCC 27853 swarms against the control (16.20 mm ± 2.55). In the anti-pyocyanin production assay, hexane fraction exhibited an inhibition of 42.66 % ± 12.94. TLC analysis and phytochemical screening revealed that hexane fraction contains steroids, terpenes, triterpenes, and glycolipids; and DCM fraction contains cardiac glycosides, triterpenoids, terpenes, triterpenes, steroids, alkaloids, and glycolipids.@*CONCLUSION@#Hexane and DCM fractions obtained from A. bilimbi significantly inhibited swarming of P. aeruginosa ATCC 27853 while none of the extracts were able to significantly inhibit pyocyanin formation of P. aeruginosa ATCC 27853.
Subject(s)
Pseudomonas aeruginosa , Averrhoa , Quorum Sensing , PyocyanineABSTRACT
@#<p style="text-align: justify;"><strong>Introduction:</strong> Nosocomial contaminants such as Pseudomonas aeruginosa and Serratia marcescens are increasingly developing resistance to many antibiotics. One of the promising alternatives that may complement, if not substitute, the use of antibiotics is quorum quenching, the process of interfering with chemical signals that mediate communication between microorganisms. Eleusine indica, a ubiquitous grass used traditionally to treat infections, has been shown to contain metabolites, such as fatty acid derivatives and p-coumaric acid, capable of quorum quenching. To date, there has been no study on the quorum quenching activity of E. indica.</p><p style="text-align: justify;"><strong>Objectives:</strong> This study aimed to determine the in vitro activity of crude ethanolic extract of E. indica leaves against selected quorum-sensing regulated virulence factors of P. aeruginosa and S. marcescens.</p><p style="text-align: justify;"><strong>Methodology:</strong> E. indica leaves were collected, washed, air-dried, and homogenized. Following ethanolic extraction and rotary evaporation, the extract was screened for antimicrobial activity through disk diffusion test and broth microdilution assay. The quorum quenching activity of the extract against P. aeruginosa was measured through swarming motility assay, while the activity against S. marcescens was measured through swarming motility and pigment inhibition assays. The quorum quenching assays were conducted in triplicates, and analysis of variance (ANOVA) was performed to identify differences among the treatment groups.</p><p style="text-align: justify;"><strong>Results:</strong> Disk diffusion test revealed that no zones of inhibition formed against both P. aeruginosa and S. marcescens for varying concentrations of up to 200 mg/mL of the crude extract. Likewise, the MIC of the extract against both P. aeruginosa and S. marcescens was determined to be >200 mg/mL. However, it was shown that the extract, at 50 mg/mL, has statistically significant activity (p<0.05) against the swarming motility of P. aeruginosa, and it is 71.6% as effective in reducing the swarming area of the bacteria compared to cinnamaldehyde. This was not observed when the extract was tested against the swarming motility of and pigment production by S. marcescens.</p><p style="text-align: justify;"><strong>Conclusion:</strong> In this study, the quorum quenching activity of the crude ethanolic extract of E. indica leaves was found to be effective against P. aeruginosa but not against S. marcescens. The compounds that will be identified by further studies may conceivably be used as an adjunct therapy in P. aeruginosa infections and as coating agents in medical devices.</p>
Subject(s)
Eleusine , Pseudomonas aeruginosa , Quorum Sensing , Serratia marcescens , ProdigiosinABSTRACT
Objective To investigate the mechanism and epidemiological characteristics of carbap-enem-resistant Proteus mirabilis ( PM) strains deficient in swarming motility. Methods PM strains were isolated from Hangzhou General Hospital of CAPF ( Chinese People′s Armed Police Forces) during January 2013 to December 2014. Bacterial motility and flagella of the PM strains were observed through semi-solid agar culture and flagella staining. Pulsed-field gel electrophoresis ( PFGE) was performed for homology anal-ysis. Antimicrobial susceptibility test and phenotypic confirmatory test were also carried out. PCR analysis and DNA sequencing were performed to confirm the genotype of resistant genes. Plasmid electroporation and S1-PFGE in combination with Southern blot hybridization were used to determine the location of the carbap-enem-resistant genes. Genetic structure of the blaKPC-2 gene was obtained by PCR mapping. Results A total of 42 PM isolates deficient in swarming motility were screened out and the resistance rates to imipenem and meropenem were 57. 1% and 52. 4%, respectively. PCR analysis and DNA sequencing confirmed that 24 carbapenem-resistant PM isolates deficient in swarming motility carried blaKPC-2 gene and belonged to three clones as indicated by the results of PFGE. Southern blot hybridization indicated that the blaKPC-2 gene was located on plasmids varying in size (26 kb, 55 kb and 139 kb). In addition, some of the strains harbored several resistant genes, such as blaTEM-1 , blaCTX-M-65 and rmtB. The genetic structures of strains carrying blaKPC-2 gene were ISKpn8, blaKPC-2 and ISKpn6-like from upstream to downstream. Conclusion Compared with the PM strains with swarming motility, the carbapenem-resistance rate was significantly higher in these PM strains deficient in swarming motility. Carbapenemases KPC-2 played an important role in the carbapen-em-resistant PM strains deficient in swarming motility. There was a cloning spread trend for carbapenem-re-sistant PM strains in our hospital. Clinicians should pay more attention to the risk of spreading.
ABSTRACT
Objective To investigate whether Pseudomonas aeruginosa pvdQ( PA2385 ) gene reveals altered antibiotic susceptibility under swarming conditions. Methods The plasmid pME6032 with pvdQ gene was constructed and identified, then transformed into Pseudomonas aeruginosa PAO1 by the electroporation, building pvdQ overexpression strain. Using the same method building pME6032-PAO1 strain.Bacteria were inoculated in LB overnight , measuring the colony diameter of the swarming zone . Results Strains of pvdQ overexpression was successfully constructed by real-time PCR. Comparison of two strains of the swarming motility of change in diameter: The result showed that PAO1 and pvdQ overexpression strains can both improve the antibiotics resistance. Swarmer cells of pvdQ overexpression strain exhibited a 2- to 4-fold increase in antibiotic resistance toward ceftazidime,ciprofloxacin, meropenem and polymyxin B compared to PAO1 on BM2-swarming agar plates. Conclusion pvdQ gene played an important role in elevating the antibiotics resistance, which through prarticipated in the swarmer cell differentiation.
ABSTRACT
Discovery of quorum sensing (QS) system to coordinate virulence and biofilm formation in bacterial pathogens has triggered search for safe, stable and non-toxic anti-QS compounds from natural products. Ethanolic extracts of 24 Indian medicinal plants were tested by agar well and disc diffusion assay for anti-QS activity using Chromobacterium violaceum (CV12472 and CVO26) reporter strains. AHL from C. violaceum CV31532 was isolated and partially purified for its use in CVO26 based bioassay. Effect on swarming-motility of Pseudomonas aeruginosa (PAO1) was also recorded at sub-MIC concentrations of extracts. Of the 24 medicinal plants screened Hemidesmus indicus (L.) Schult (root), Holarrhena antidysenterica (Roth)A.DC. (bark), Mangifera indica L. (seed) Punica granatum L. (pericarp) and Psoralea corylifolia L. (seed) demonstrated varying level of inhibition of violacein production in the reporter strains. Moreover, a significant reduction in swarms was recorded over control. The inhibition of violacein production and swarming motility may be due to direct or indirect interference on QS by active constituents or the interactive effect of different phytocompounds present in the extracts. These plant extracts may be selected for activity guided fractionation to identify and characterize the active principle
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Objective To investigate the periodic variety at the germ form, quantity, flagellar expression and activity of breath enzyme etc during the swarming motility of Proteus mirabilis (PM). Methods The 2, 3, 5-tetraphenyltetrazolium chloride (TTC), as indictor of breath enzyme, was added to the culture medium. Electronic microscope was adopted to observe the germ form and flagellar. Quantity of PM was measured by turbidimetry. Results During the PM growth and migration at LB medium, the germ quantity reduced and form changed from short and small to long and thin, flagellar got more and breath enzyme turned into inactive. Conclusion At the LB medium, PM in growth process appeared varieties in length of germ body, flagellar quantity and germs density, etc. PM in solid medium appeared wave motility, resulting from many factors including the environment, the signal transduction in cells and the density of germ.
ABSTRACT
Objective To study the molecular mechanism of swarming motility in Citrobacter freundii.Methods Mini-Tn5 transposon was used to induce swarming mutants in Citrobacter freundii.The mutant genes were amplified by inverse-PCR and were then sequenced.Mutants were observed under light microscope and electron microscope to determine the characters of motility and bacterial shape.Results Seven swarming mutants were found to be three gene mutants(wzxB,waaW,waaL) involving LPS synthesis.Moreover,flagellar production of these mutants was abnormal,that is,only small portion of bacterial cells produced flagella normally.The majority of cells tended to aggregate and produced little flagella observed under electron microscope.Conclusion Absence of O-antigen affects the production of flagella and swarming motility of Citrobacter(freundii).