Clostridium perfringens TpeL belongs to a family of large clostridial cytotoxins that encompasses Clostridium difficile toxin A (TcdA) and B (TcdB) and Clostridium sordellii lethal toxin (TcsL). We report here the identification of the TpeL-catalyzed modification of small GTPases. A recombinant protein (TpeL1-525) derived from the TpeL N-terminal catalytic domain in the presence of streptolysin O (SLO) induced the rounding of Vero cells and the glycosylation of cellular Rac1.
View Article and Find Full Text PDFFour pasture-fed Japanese Black cows showed the main clinical symptoms of severe hemoglobinuria at different periods between 2003 and 2007. Hematological analyses at the first consultation revealed severe anemia, and biochemical analyses indicated both severe hemolysis and disruption of hepatic function. Although the first 2 patients died, the hemoglobinuria and general condition of the remaining 2 cows, who were immediately initiated on large doses of antibiotics, improved within 3 days.
View Article and Find Full Text PDFThe objectives of this study were to identify haemagglutinin (HA) epitopes of Avibacterium paragallinarum serovar C that are capable of eliciting haemagglutination inhibition (HI) antibody, and to investigate their immunogenic role. Three conformational epitopes were detected on HA by blocking ELISA and immuno-dot blot analysis using a panel of five monoclonal antibodies (MAbs) with HI activity, designated 8C1C, 4G8B, 24E4D, 11E11B, and 10D1A. The minimum DNA regions coding these three epitopes were 3195, 2862, and 807bp in size, and mapped within a gene with 6117bp.
View Article and Find Full Text PDFThe aims of this study were the identification, cloning, and expression of a genetic region encoding an epitope that induces hemagglutination inhibition (HI) antibody against Avibacterium paragallinarum serovar A and an evaluation of the recombinant protein for immunogenicity in chickens. Although two monoclonal antibodies (MAbs) with HI activity, designated S24-951 and S7-1716-5C, were generated in this study, no reactive proteins with both MAbs were identified by Western blot analysis. A gene fragment of 5157 bp, designated hpa5.
View Article and Find Full Text PDFAn unknown cytotoxin was identified in the culture supernatant of Clostridium perfringens type C. The cytotoxin, named TpeL, which was purified using mAb-based affinity chromatography, had a lethal activity of 62 minimum lethal dose (MLD) mg(-1) in mice and a cytotoxic activity of 6.2x10(5) cytotoxic units (CU) mg(-1) in Vero cells.
View Article and Find Full Text PDFClostridium septicum alpha-toxin genes were sequenced with the polymerase chain reaction (PCR) products amplified from DNAs of 25 C. septicum strains, and were classified into 10 patterns. Alpha-toxins were purified from the culture supernatant of four C.
View Article and Find Full Text PDFPasteurella multocida, serovar A: 3, selected by pathogenicity in mice from among 10 strains isolated from pneumonic lesions of calves, was adjusted to 10(7), 10(8) and 10(10) colony-forming units (CFU), and inoculated intratracheally into four calves. All calves showed pyrexia and had lungs with congestion and hepatization. Inoculation with 10(10) CFU of bacteria produced respiratory symptoms and abscesses in lungs.
View Article and Find Full Text PDFThe protective effect of an alpha-toxoid vaccine of Clostridium septicum purified alpha-toxin was investigated in guinea pigs. Purified alpha-toxin was treated with formalin to make toxoid, and alpha-toxoid vaccine was prepared by mixing alpha-toxoid (4 to 64 microg/dose) with an aluminum phosphate gel as adjuvant. Guinea pigs were immunized twice with different doses of alpha-toxoid vaccine, and challenged with spores of C.
View Article and Find Full Text PDF