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      Effects of norspermidine on Pseudomonas aeruginosa biofilm formation and eradication

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          Abstract

          Biofilms are defined as aggregation of single cell microorganisms and associated with over 80% of all the microbial infections. Pseudomonas aeruginosa is a Gram‐negative opportunistic pathogen capable of leading to various infections in immunocompromised people. Recent studies showed that norspermidine, a kind of polyamine, prevented and disrupted biofilm formation by some Gram‐negative bacterium. In this study, the effects of norspermidine on P. aeruginosa biofilm formation and eradication were tested. Microtiter plate combined with crystal violet staining was used to study the effects of norspermidine on P. aeruginosa initial attachment, then we employed SEM (scanning electron microscope), qRT‐PCR, and QS‐related virulence factor assays to investigate how norspermidine prevent biofilm formation by P. aeruginosa. We reported that high‐dose norspermidine had bactericide effect on P. aeruginosa, and norspermidine began to inhibit biofilm formation and eradicate 24‐h mature biofilm at concentration of 0.1 and 1 mmol/L, respectively, probably by preventing cell‐surface attachment, inhibiting swimming motility, and downregulating QS‐related genes expression. To investigate the potential utility of norspermidine in preventing device‐related infections, we found that catheters immersed with norspermidine were effective in eradicating mature biofilm. These results suggest that norspermidine could be a potent antibiofilm agent for formulating strategies against P. aeruginosa biofilm.

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          Most cited references41

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          Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development.

          The formation of complex bacterial communities known as biofilms begins with the interaction of planktonic cells with a surface in response to appropriate environmental signals. We report the isolation and characterization of mutants of Pseudomonas aeruginosa PA14 defective in the initiation of biofilm formation on an abiotic surface, polyvinylchloride (PVC) plastic. These mutants are designated surface attachment defective (sad ). Two classes of sad mutants were analysed: (i) mutants defective in flagellar-mediated motility and (ii) mutants defective in biogenesis of the polar-localized type IV pili. We followed the development of the biofilm formed by the wild type over 8 h using phase-contrast microscopy. The wild-type strain first formed a monolayer of cells on the abiotic surface, followed by the appearance of microcolonies that were dispersed throughout the monolayer of cells. Using time-lapse microscopy, we present evidence that microcolonies form by aggregation of cells present in the monolayer. As observed with the wild type, strains with mutations in genes required for the synthesis of type IV pili formed a monolayer of cells on the PVC plastic. However, in contrast to the wild-type strain, the type IV pili mutants did not develop microcolonies over the course of the experiments, suggesting that these structures play an important role in microcolony formation. Very few cells of a non-motile strain (carrying a mutation in flgK) attached to PVC even after 8 h of incubation, suggesting a role for flagella and/or motility in the initial cell-to-surface interactions. The phenotype of these mutants thus allows us to initiate the dissection of the developmental pathway leading to biofilm formation.
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            Common virulence factors for bacterial pathogenicity in plants and animals.

            A Pseudomonas aeruginosa strain (UCBPP-PA14) is infectious both in an Arabidopsis thaliana leaf infiltration model and in a mouse full-thickness skin burn model. UCBPP-PA14 exhibits ecotype specificity for Arabidopsis, causing a range of symptoms from none to severe in four different ecotypes. In the mouse model, UCBPP-PA14 is as lethal as other well-studied P. aeruginosa strains. Mutations in the UCBPP-PA14 toxA, plcS, and gacA genes resulted in a significant reduction in pathogenicity in both hosts, indicating that these genes encode virulence factors required for the full expression of pathogenicity in both plants and animals.
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              Type IV pili and twitching motility.

              Twitching motility is a flagella-independent form of bacterial translocation over moist surfaces. It occurs by the extension, tethering, and then retraction of polar type IV pili, which operate in a manner similar to a grappling hook. Twitching motility is equivalent to social gliding motility in Myxococcus xanthus and is important in host colonization by a wide range of plant and animal pathogens, as well as in the formation of biofilms and fruiting bodies. The biogenesis and function of type IV pili is controlled by a large number of genes, almost 40 of which have been identified in Pseudomonas aeruginosa. A number of genes required for pili assembly are homologous to genes involved in type II protein secretion and competence for DNA uptake, suggesting that these systems share a common architecture. Twitching motility is also controlled by a range of signal transduction systems, including two-component sensor-regulators and a complex chemosensory system.
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                Author and article information

                Journal
                Microbiologyopen
                Microbiologyopen
                10.1002/(ISSN)2045-8827
                MBO3
                MicrobiologyOpen
                John Wiley and Sons Inc. (Hoboken )
                2045-8827
                27 January 2016
                June 2016
                : 5
                : 3 ( doiID: 10.1002/mbo3.2016.5.issue-3 )
                : 402-412
                Affiliations
                [ 1 ] Department of Medicine Clinical LaboratoryThe Third Xiangya Hospital of Central South University Changsha 410013China
                [ 2 ] Xiangya School of MedicineCentral South University Changsha 410013China
                Author notes
                [*] [* ] Correspondence

                Yong Wu, Department of Medicine Clinical Laboratory, The Third Xiangya Hospital of Central South University, Changsha 410013, China. Tel: +86 731 8861 8441; Fax: +86 731 8861 8441; E‐mail: wuyong_xy@ 123456126.com

                [†]

                These authors contributed equally to this work.

                Article
                MBO3338
                10.1002/mbo3.338
                4905993
                26817804
                4739df47-acc3-4085-abb5-27574c28095a
                © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 29 September 2015
                : 15 December 2015
                : 27 December 2015
                Page count
                Pages: 11
                Funding
                Funded by: Natural Science Foundation of Hunan Province
                Award ID: 12JJ3092
                Award ID: 2015JJ2188
                Award ID: 2015JJ3165
                Funded by: Graduate Student Innovation Project of the Central South University
                Award ID: 502200714
                Categories
                Original Research
                Original Research
                Custom metadata
                2.0
                mbo3338
                June 2016
                Converter:WILEY_ML3GV2_TO_NLMPMC version:4.9.1 mode:remove_FC converted:13.06.2016

                Microbiology & Virology
                bacterial adhesion,biofilm,norspermidine,pseudomonas aeruginosa,quorum sensing

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