F and type 1 piliation of various Escherichia coli K12 Hfr and F + strains was re-examined by using a new visualization assay. In anaerobic cultures in rich media, bacteria were well F piliated throughout all growth phases. In aerobic cultures in rich media, F piliation reached a maximum in the mid-exponential phase. The yield of pili was up to 15 mg 1 -l . In aerated cultures, F pili disappeared in the late exponential phase. In rich media F pili were on average 10-20 pm long; in synthetic media they were an order of magnitude shorter, and less numerous. Addition of metabolic poisons (NaCN, NaN,, Na3As0,, phenylethyl alcohol) and starving the bacteria caused rapid disappearance of F pili under aerobic conditions, but had no influence on F piliation under anaerobic conditions. Type 1 piliation was not influenced by these drugs or by an alteration of growth conditions.
Methods for the direct visualization of F and type 1 pili of Escherichia coli in the light microscope are described. The method for visualizing F pili is based on the specific adsorption of fluorescent dye-labelled RNA phages to F pili. The best results were obtained with MS2 phages labelled with rhodamine B. Semi-quantitative determination of the amount of F pili is possible. Type 1 pili can be visualized rapidly and specifically by indirect immunofluorescence. Other structures on the cell surface are neither detected by, nor interfere with these assays. By using different fluorescent dyes the two methods can be combined and both F and type 1 pili can be determined in the same sample.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.