Detection of Vibrio (Listonella) anguillarum Porin Homologue Proteins and Their Source Bacteria from Coastal Seawater

Abstract

The molecular distribution of dissolved proteins in seawater from coastal marine environments in Uranouchi Bay, Kochi Prefecture, is first reported in this article. Occurrence of bacteria-derived dissolved proteins and their source bacteria were examined using a probe of the antibody (anti-Omp35La) against a porin outer membrane protein (Omp35La) of the fish pathogenic bacterium Vibrio (Listonella) anguillarum. The electrophoretograms of dissolved proteins from coastal seawater showed a large number of discrete and individual proteins overlapped each other over a wide range of molecular masses indicating active processes in coastal environments in transferring proteins from organisms to the inanimate dissolved protein pool. Among the dissolved proteins, 37 kDa- and 18 kDa-proteins reacted with the Omp35La. In order to isolate the source bacteria of such dissolved proteins, bacteria from seawater and diseased fish were screened by colony Western blotting with anti-Omp35La. The reactive strains were further examined in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blotting to verify the presence of Omp35La homologues among the outer membrane proteins of such strains. Outer membrane proteins reacting with anti-Omp35La were detected in only 4 strains of the 129 strains that were positive in the colony Western blotting. The level of possible source bacteria of 37 kDa- and 18 kDa-dissolved proteins was suggested to be 5–6 orders of magnitude lower than the total bacterial count. The present study leads us to hypothesize that a minor portion of the bacterial assemblage is responsible for the dissolved proteins in the coastal waters.