In order to investigate the main factor of pathogenicity in V. mimicus, we have studied the toxic effects of j3-hemolysin produced by V. mimicus. The purified hemolysin of V. mimicus was active erythrocytes from three animal species including mouse, rabbit and rat, but the hemolysin was most active against rabbit erythrocyte. The hemolysin lysed cultured cell and killed mouse. Rapid death of mouse was observed with rather small doses of the toxin. Intravenous injection of 20 mg of the purified toxin killed mice within 25 sec. The hemolysin also had a lethal effect on intraperitoneal injection into mice although less than on intravenous injection. Purified hemolysin injected rabbits had large morphological change in jejunum. In electron micrograph of thin sections of the human erythrocytes, cells were threated with the hemolysin at 37 C for 5 min., significant changes were not observed. But after 10 min., hemolysis was observed and after 60 min., complete degradation of human erythrocyte was observed.
Animals ; Cells, Cultured ; Erythrocytes ; Hemolysis ; Humans ; Injections, Intraperitoneal ; Injections, Intravenous ; Jejunum ; Mice ; Rabbits ; Rats ; Vibrio mimicus* ; Vibrio* ; Virulence

Growth under conditions of iron-restriction and the production of siderophore was examined in Vibrio mimicus ATCC 33653. This strain grew and multiplied in the presence of the high-affinity iron chelators ethylenediamine-di (o-hydroxyphenylacetic acid). Chrorne azurol S (CAS) agar and solution were used to detect the production of siderophore under these condition. Siderophore could be detected in the iron-rcstricted culture supernatants. The siderophore was extracted from iron-restricted culture supernatants by phenol-chloroform-ether method and purified by Dowex ion-exchange and Sephadex G-25 gel filtracton chromatography. The purified siderophore was confirmed by paper chromatography and HPLC. The Purified siderophore enhanced the growth of V. mimicus when the bacterium was grown in iron limited medium. Injection of both the siderohore and the bacteria to mice resulted in more rapid death than that of the only bacteria. However, the siderophore did not show lethality to mice and any toxicity to cell line like HeLa and U937.
Agar ; Animals ; Bacteria ; Cell Line ; Chelating Agents ; Chromatography ; Chromatography, High Pressure Liquid ; Chromatography, Paper ; Iron ; Mice ; Vibrio mimicus* ; Vibrio*

Vibrio mimicus, marine bacteria pathogenic for fish, can causes acute gastroenteritis in human. Iron limmited condition like in human body, may change the surface structure of V. mimicus. In this study we obse'rved the effect of iron limmited condition on outer membrane protein of V. mimicus. Ethylenediamine-di (O-hydroxy-phenylacetic) acid (EDDA), an iron chelator, delayed the time to reach expotential growth of V. mimicus in brain heart infusion medium from 3 hours to 20 hours. Outer membrane protein of V. mimicus-CON (cultured in BHI) and V. mimicus-EDDA (cultured in BHI contain EDDA) were seperated by 1% sarcosine from total cell envelop. SDS-PAGE of V. mimicus-EDDA and V. mimicus-CON showed similar protein profiles contain 37 kDa major protein but 86 and 90 kDa protein were induced differently. Immunological properties of above protein were determined by ELISA and western blotting. 86 kDa EDDA- specific OMP was induced in V. mimicus (isolate 96-1), V. parahaemolyticus (serotype 09), V. alginolyticus (isolate 95-1), E. coli (human isolate) and V. vulnificus ATCC 27562 in iron limmited condition.
Bacteria ; Blotting, Western ; Brain ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gastroenteritis ; Heart ; Human Body ; Humans ; Iron* ; Membrane Proteins ; Membranes* ; Sarcosine ; Vibrio mimicus* ; Vibrio*