Bile Salt-induced Biofilm Formation in Enteric Pathogens: Techniques for Identification and Quantification

Journal of Visualized Experiments : JoVE
Kourtney P Nickerson, Christina S Faherty

Abstract

Biofilm formation is a dynamic, multistage process that occurs in bacteria under harsh environmental conditions or times of stress. For enteric pathogens, a significant stress response is induced during gastrointestinal transit and upon bile exposure, a normal component of human digestion. To overcome the bactericidal effects of bile, many enteric pathogens form a biofilm hypothesized to permit survival when transiting through the small intestine. Here we present methodologies to define biofilm formation through solid-phase adherence assays as well as extracellular polymeric substance (EPS) matrix detection and visualization. Furthermore, biofilm dispersion assessment is presented to mimic the analysis of events triggering release of bacteria during the infection process. Crystal violet staining is used to detect adherent bacteria in a high-throughput 96-well plate adherence assay. EPS production assessment is determined by two assays, namely microscopy staining of the EPS matrix and semi-quantitative analysis with a fluorescently-conjugated polysaccharide binding lectin. Finally, biofilm dispersion is measured through colony counts and plating. Positive data from multiple assays support the characterization of biofilms and can...Continue Reading

Citations

Nov 15, 2019·MSphere·Rachael B ChaninChristina S Faherty
Jul 15, 2020·Infection and Immunity·Kevin S GipsonChristina S Faherty
Sep 21, 2021·Journal of Bacteriology·Ruchi OjhaBenjamin J Koestler

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biofilms

Biofilms are adherent bacterial communities embedded in a polymer matrix and can cause persistent human infections that are highly resistant to antibiotics. Discover the latest research on Biofilms here.

Biofilm & Infectious Disease

Biofilm formation is a key virulence factor for a wide range of microorganisms that cause chronic infections.Here is the latest research on biofilm and infectious diseases.