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Activity of Norspermidine on Bacterial Biofilms of Multidrug-Resistant Clinical Isolates Associated with Persistent Extremity Wound Infections

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Book cover Advances in Microbiology, Infectious Diseases and Public Health

Part of the book series: Advances in Experimental Medicine and Biology ((AMIDPH,volume 973))

Abstract

Biofilm formation is a major virulence factor for numerous pathogenic bacteria and is cited as a central event in the pathogenesis of chronic human infections, which is in large part due to excessive extracellular matrix secretion and metabolic changes that occur within the biofilm rendering them highly tolerant to antimicrobial treatments. Polyamines, including norspermidine, play central roles in bacterial biofilm development, but have also recently been shown to inhibit biofilm formation in select strains of various pathogenic bacteria. The aim of this study was to evaluate in vitro the biofilm dispersive and inhibitory activities of norspermidine against multidrug-resistant clinical isolates of Acinetobacter baumannii(n = 4), Klebsiella pneumoniae (n = 3), Pseudomonas aeruginosa (n = 5) and Staphylococcus aureus (n = 4) associated with chronic extremity wound infections using the semi-quantitative 96-well plate method and confocal laser microscopy. In addition to the antibiofilm activity, biocompatibility of norspermidine was also evaluated by measuring toxicity in vitro to human cell lines and whole porcine tissue explants using MTT viability assay and histological analysis. Norspermidine (5–20 mM) had variable dispersive and inhibitory activity on biofilms which was dependent on both the strain and species. Of the clinical bacterial species evaluated herein, A. baumannii isolates were the most sensitive to the effect of norspermidine, which was in part due to the inhibitory effects of norspermidine on bacterial motility and expression of genes involved in the production of homoserine lactones and quorum sensing molecules both essential for biofilm formation. Importantly, exposure of cell lines and whole tissues to norspermidine for prolonged periods of time (≥24 h) was observed to reduce viability and alter tissue histology in a time and concentration dependent manner, with 20 mM exposure having the greatest negative effects on both tissues and individual cell lines. Collectively our findings demonstrate that, similar to other polyamines, norspermidine displays both inhibitory and dispersive activities on biofilms of clinical multidrug-resistant bacterial isolates, in particular for strains of A. baumannii. Additionally our findings suggest that direct application may be considered on tissues, albeit for limited exposure times.

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Acknowledgments

This work was supported by intramural funding from the Combat Casualty Research Program, Medical Research and Materiel Command. Additional support for this work (IDCRP-024) was provided by in part by the Infectious Disease Clinical Research Program (IDCRP), a Department of Defense program executed through the Uniformed Services University of the Health Sciences. This project has been funded by the Department of Defense Global Emerging Infections Surveillance and Response System (GEIS), a division of the Armed Forces Health Surveillance Center, National Institute of Allergy and Infectious Diseases, National Institute of Health, under Inter-Agency Agreement Y1-AI-5072, and the Department of the Navy under the Wounded, Ill, and Injured Program. We are indebted to the Infectious Disease Clinical Research Program TIDOS study team. Lastly we would also like to thank Kinton Armmer and Douglas Cortez for their assistance with isolation and preparation of the porcine tissues used in this study.

Competing Interests

The authors declare that they have no competing interests.

Disclaimer

The opinions or assertions contained herein are the private views of the authors are not to be construed as official or as reflecting the views of the Uniformed Services University of the Health Sciences, the National Institute of Health or the Department of Health and Human Services, the Department of Defense, the Departments of the Army, Navy or Air Force, or the United States Army Institute of Surgical Research. Mention of trade names, commercial products, or organization does not imply endorsement by the U.S. Government.

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Authors and Affiliations

  1. Infectious Disease Service, Department of Medicine, San Antonio Military Medical Center, JBSA Fort Sam Houston, San Antonio, TX, USA

    Anthony P. Cardile, Katrin Mende & Kevin S. Akers

  2. Extremity Trauma and Regenerative Medicine Task Area, United States Army Institute of Surgical Research, JBSA Fort Sam Houston, San Antonio, TX, USA

    Ronald L. Woodbury, Carlos J. Sanchez Jr, Sandra C. Becerra, Rebecca A. Garcia, Joseph C. Wenke & Kevin S. Akers

  3. Infectious Disease Clinical Research Program, Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, USA

    Katrin Mende

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  1. Anthony P. Cardile
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Correspondence to Kevin S. Akers .

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  1. Microbial Biofilm Laboratory, Fondazione Santa Lucia IRCCS, Rome, Italy

    Gianfranco Donelli

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Authors’ Contributions

APC, RLW and CJS conceived and designed the experiments. RLW, SCB, RAG and CJS performed the experiments. APC and CJS analyzed the data. APC, KM, KSA, JCW and CJS contributed to preparation and review of the final manuscript. All authors read and approved the final manuscript.

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Cardile, A.P. et al. (2016). Activity of Norspermidine on Bacterial Biofilms of Multidrug-Resistant Clinical Isolates Associated with Persistent Extremity Wound Infections. In: Donelli, G. (eds) Advances in Microbiology, Infectious Diseases and Public Health. Advances in Experimental Medicine and Biology(), vol 973. Springer, Cham. https://doi.org/10.1007/5584_2016_93

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