caa a22 4500 477105971 CHVBK 20180405111553.0 cr unu---uuuuu 170330e19960601xx s 000 0 eng 10.1007/BF01570111 doi (NATIONALLICENCE)springer-10.1007/BF01570111 Tracing the interaction of bacteriophage with bacterial biofilms using fluorescent and chromogenic probes [Elektronische Daten] [M Doolittle, J Cooney, D Caldwell] Phages T4 and E79 were fluorescently-labeled with rhodamine isothiocyanate (RITC), fluoroscein isothiccyanate (FITC), and by the addition of 4′6-diamidino-2-phenylindole (DAPI) to phage-infected host cells ofEscherichia coli andPseudomonas aeruginosa. Comparisons of electron micrographs with scanning confocal laser microscope (SCLM) images indicated that single RITC-labeled phage particles could be visualized. Biofilms of each bacterium were infected by labeled phage. SCLM and epifluorescence microscopy were used to observe adsorption of phage to single-layer surface-attached bacteria and thicker biofilms. The spread of the recombinant T4 phage, YZA1 (containing an rll-LacZ fusion), within alac E. coli biofilm could be detected in the presence of chromogenic and fluorogenic homologs of galactose. Infected cells exhibited blue pigmentation and fluorescence from the cleavage products produced by the phage-encoded β-galactosidase activity. Fluorescent antibodies were used to detect nonlabeled progeny phage. Phage T4 infected both surface-attached and surface-associatedE. coli while phage E79 adsorbed toP. aeruginosa cells on the surface of the biofilm, but access to cells deep in biofilms was somewhat restricted. Temperature and nutrient concentration did not affect susceptibility to phage infection, but lower temperature and low nutrients extended the time-to-lysis and slowed the spread of infection within the biofilm. Society for Industrial Microbiology, 1996 biofilms nationallicence bacteriophages T4 and E79 nationallicence Escherichia coli nationallicence Pseudomonas aeruginosa nationallicence SCLM nationallicence fluorescence nationallicence Doolittle M. Environmental Sciences Program, University of Massachusetts, 02125-3393, Boston, MA, USA aut Cooney J. Environmental Sciences Program, University of Massachusetts, 02125-3393, Boston, MA, USA aut Caldwell D. Department of Applied Microbiology and Food Science, University of Saskatchewan, S7N OWO, Saskatoon, Canada aut Journal of Industrial Microbiology Springer-Verlag 16/6(1996-06-01), 331-341 0169-4146 16:6<331 1996 16 10295 https://doi.org/10.1007/BF01570111 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01570111 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Doolittle M. Environmental Sciences Program, University of Massachusetts, 02125-3393, Boston, MA, USA aut NATIONALLICENCE 700 1- Cooney J. Environmental Sciences Program, University of Massachusetts, 02125-3393, Boston, MA, USA aut NATIONALLICENCE 700 1- Caldwell D. Department of Applied Microbiology and Food Science, University of Saskatchewan, S7N OWO, Saskatoon, Canada aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology Springer-Verlag 16/6(1996-06-01), 331-341 0169-4146 16:6<331 1996 16 10295 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer