Open AccessVibrios adhere to epithelial cells in the intestinal tract of red sea bream, P agrus major, utilizing GM 4 as an attachment site
Author(s) -
Chisada Shinichi,
Shimizu Kohei,
Kamada Haruna,
Matsunaga Naoyuki,
Okino Nozomu,
Ito Makoto
Publication year - 2013
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12082
Subject(s) - pagrus major , microbiology and biotechnology , vibrio harveyi , vibrio , biology , antibody , vibrionaceae , chemistry , bacteria , fish <actinopterygii> , fishery , immunology , genetics
Vibrios, distributed in marine and brackish environments, can cause vibriosis in fish and shellfish under appropriate conditions. Previously, we clarified by thin‐layer chromatography ( TLC ) overlay assay that 35 S‐labeled Vibrio trachuri adhered to GM 4 isolated from red sea bream intestine. However, whether GM 4 actually functions on epithelial cells as an attachment site for vibrios still remains to be uncovered. We found that six isolates, classified as V. harveyi , V. campbellii , and V. splendidus, from intestinal microflora of red sea bream adhered to GM 4 but not galactosylceramide (GalCer) by TLC ‐overlay assay. Tissue‐overlay assays revealed that V. harveyi labeled with green fluorescent protein ( GFP ) adhered to epithelial cells of red sea bream intestine where GM 4 and GalCer were found to be distributed on the top layer of actin filaments by immunohistochemical analysis using corresponding antibodies. The number of adhering vibrios was diminished by pretreatment with anti‐ GM 4 antibody, but not anti‐GalCer antibody. These results clearly indicate that vibrios adhere to epithelial cells of red sea bream intestine utilizing GM 4 as an attachment site.