• Korean Journal of Veterinary Research
• /
• v.57 no.1
• /
• pp.9-15
• /
• 2017

To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.

• Proceedings of the Microbiological Society of Korea Conference
• /
• 2004.05a
• /
• pp.147-149
• /
• 2004

• Asian-Australasian Journal of Animal Sciences
• /
• v.9 no.6
• /
• pp.671-677
• /
• 1996

Erythrocytes from three different animal species were used to determine mannose-sensitive hemagglutination (MSHA) and mannose-resistant hemagglutination (MRHA) of 755 isolates obtained from rectal swabs of healthy pig. In addition, colony hybridization using digoxigenin-dUTP labeled polynucleotide probes was performed for the detection of heat-stable and heat-labile enterotoxin genes carried by MRHA positive isolates. Of 755 strains, 9, 4 and 28 strains gave a positive MRHA with bovine, equine and pig erythrocytes, respectively. Of these isolates, 28 (3.7%) were characterized for positive MRHA by at least one blood. Seven isolates gave a positive MRHA with two kinds of blood. Three gave a positive MRHA with three kinds of blood. Twenty-eight strains, while positive in MRHA, yielded negative signals in the colony hybridization assay for the detection of heat-stable (STaI and STaII) and heat-labile (LT) enterotoxin genes in E. coli.

• The Journal of the Korean Society for Microbiology
• /
• v.22 no.4
• /
• pp.377-392
• /
• 1987

Monoclonal antibody to the Escherichia coli heat-stable enterotoxin(ST) was produced to develop a rapid and convenient diagnostic method to the ST. The toxin was purified from culture supernatant of enterotoxigenic E. coli O148H28($ST^+/LT^+$) and conjugated to bovine serum albumin(BSA). The ST-BSA conjugate was used to immunize BALB/c mice and the immune spleen cells from these mice were fused with $P3{\times}63$ Ag8.V653 plasmacytoma cells. Hybridomas were screened by ELISA and positive hybridomas were cloned by limiting dilution. Finally, one stable clone (AS36) specific to ST was selected for further growth and characterization. Antibody titers of culture supernatant and ascitic fluid from BALB/c mice were 1:1,024 and 1:20,480 respectively in ELISA. The isotype and subclass of monoclonal antibody was IgG1 in sandwich ELISA. To test the neutralizing effect of monoclonal antibody on toxin activity of ST, mixture of ascitic fluid and ST was assayed by infant mouse assay and this monoclonel antibody was proved to be a neutralizing antibody. The titer of ascitic fluid which completely neutralized biological activity of 4 units of ST was 1:4. Purified ST was quantitatively measured by competitive ELISA and minimum amount of ST detectable by this assay was 250pg, which was an amount six-fold smaller than that detectable by infant mouse assay. Four reference strains of enterotoxigenic E. coli from WHO were detected by competitive ELISA and highly specific, sensitive and reproducible result was obtained.