Investigation of Biofilm Formation in Yersinia pestis
The formation of a biofilm, which often grows as an attached extracellular matrix and blocks the flea proventriculus, enhances flea-borne transmission of Yersinia pestis during flea biting. Quantitative investigation of biofilm formation can be used to evaluate Y. pestis transmission in vitro. This chapter reviews methods used to investigate Y. pestis biofilm formation. These methods can be divided into three types based on (i) direct observation of biofilm formation including biofilm colony, crystal violet staining, and Caenorhabditis elegans biofilm assay, (ii) quantitative evaluation of the expression of genes related to biofilm formation by real-time polymerase chain reaction, and (iii) quantification of intracellular cyclic diguanylic acid (c-di-GMP) concentration, which positively correlates with Yersinia biofilm formation.
Key wordsCrystal violet staining Biofilm formation c-di-GMP Caenorhabditis elegans