title: Use of a stainless steel washer platform to study Acinetobacter baumannii adhesion and biofilm formation on abiotic surfaces.
abstract: Acinetobacter baumannii is a frequent cause of hospital-acquired pneumonia, and has recently increased in incidence as the causative agent of severe disease in troops wounded in Afghanistan and Iraq. Clinical approaches are limited since A. baumannii strains isolated from patients are extremely resistant to current antimicrobials. A. baumannii can survive desiccation and during outbreaks has been recovered from various sites in the patients' environment. To better understand its prevalence in hospital settings, we used a stainless steel washer (SSW) platform to investigate A. baumannii biofilm formation on abiotic surfaces. Scanning electron microscopy demonstrated that A. baumannii forms strong biofilms on stainless steel surfaces. This platform was combined with a colorimetric 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay to observe the metabolic activity of bacterial cells, and to facilitate the manipulation and comparison of multiple A. baumannii clinical strains. A strong correlation between XTT and c.f.u. assays was demonstrated. To complement the cell viability assays, A. baumannii biofilm mass was measured by crystal violet staining. Furthermore, the effect of commonly used disinfectants and environmental stressors on A. baumannii biofilms and planktonic cells was compared and characterized. Biofilms on SSWs were significantly more resistant than their planktonic counterparts, providing additional evidence that may allow us to understand the high prevalence of this microbe in hospital settings. Our results validate that SSWs are a simple, versatile and innovative method to study A. baumannii biofilms in vitro.
authors.affiliation: Department of Biomedical Sciences, Long Island University-Post, Brookville, NY, USA.
authors.forename: Samantha J
authors.initials: SJ
authors.lastname: Orsinger-Jacobsen
authors.affiliation: Department of Biomedical Sciences, Long Island University-Post, Brookville, NY, USA.
authors.forename: Shenan S
authors.initials: SS
authors.lastname: Patel
authors.affiliation: Department of Biomedical Sciences, Long Island University-Post, Brookville, NY, USA.
authors.forename: Ernestine M
authors.initials: EM
authors.lastname: Vellozzi
authors.affiliation: Microbiology Laboratory, Montefiore Medical Center, Bronx, NY, USA.
authors.forename: Phillip
authors.initials: P
authors.lastname: Gialanella
authors.affiliation: Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Nimrichter
authors.affiliation: Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Kildare
authors.initials: K
authors.lastname: Miranda
authors.affiliation: Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, USA.|Departments of Medicine, Division of Infectious Diseases, Albert Einstein College of Medicine, Bronx, NY, USA.|Department of Biomedical Sciences, Long Island University-Post, Brookville, NY, USA.
authors.forename: Luis R
authors.initials: LR
authors.lastname: Martinez
mesh_terms: D040981:Acinetobacter baumannii / Q000502:physiology*; D000346:Afghanistan; D001422:Bacterial Adhesion; D018441:Biofilms / Q000254:growth & development*; D015169:Colony Count, Microbial; D003124:Colorimetry; D004783:Environmental Microbiology; D005840:Gentian Violet / Q000378:metabolism; D006801:Humans; D007493:Iraq; D050296:Microbial Viability; D008855:Microscopy, Electron, Scanning; D010084:Oxidation-Reduction; D013194:Staining and Labeling; D013193:Stainless Steel; D013778:Tetrazolium Salts / Q000378:metabolism; D014481:United States; D014946:Wound Infection / Q000382:microbiology
chemical_list: D013778:Tetrazolium Salts; C059087:2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide; D013193:Stainless Steel; D005840:Gentian Violet
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