Biotic and abiotic substrates for enhancing Acinetobacter baumannii biofilm formation: New approach using extracellular matrix and slanted coverslip technique

• Published in: Journal of general and applied microbiology Vol. 65; no. 2; pp. 64 - 71
• Main Authors: Ramalingam, Karthikeyan, Lee, Valerie
• Format: Journal Article
• Language: English
• Published: Tokyo Applied Microbiology, Molecular and Cellular Biosciences Research Foundation 01-01-2019; Japan Science and Technology Agency
• Subjects: Acinetobacter baumannii; Assaying; biofilm growth; biofilm substrates; Biofilms; Extracellular matrix; Glass; Inserts; Pili; Substrates
• ISSN: 0022-1260; 1349-8037
• DOI: 10.2323/jgam.2018.05.004

Acinetobacter baumannii has been well recognized as a problematic human pathogen and several reports has shown the incidence of multidrug and pandrug-resistant A. baumannii strains in infirmary infections. A. baumannii grows only on an air-liquid interface and does not form a contiguous biofilm. Extracellular matrices (ECM) and slanted glass coverslips are (SGC) used as biofilm substrates and biofilms have been investigated by SEM, confocal and crystal violet staining. ECM has shown enhanced biofilm formation under dynamic conditions rather than static conditions. SGC biofilm yield assay has shown higher levels of continuous layers and packed thicker biofilm formation with glass coverslip inserts, up to 1.7 to 3 times higher biofilm formation, than when compared with no glass coverslip inserts. A media immersed ECM study revealed that biofilm grown on extracellular matrixes formed thread-like pili structures, and that these structures had contact with the ECM and also showed excellent cell-to-cell interaction. In summary, A. baumannii showed higher biofilm formation capacities with ECM, while the prominent results were directly related with the biofilm formation capacity of A. baumannii. For the initial step of biofilm formation, adherence is an important factor and, consequently, strains with a comparatively high capability to adhere to extracellular matrices and slanted glass coverslips provide a new method of enhanced biofilm growth for in vitro assays. ECM can be used as a substrate for immersed biofilm formation studies and the SGC method for air-liquid interface exposed biofilm formation studies, and these substrates can provide better biofilm growth and easy handling for in vitro adherence and biofilm assays.