Enterotoxin formation by Clostridium perfringens type A studied by the use of fluorescent antibody

Fluorescein isothiocyanate-conjugated antibody to purified enterotoxin of Clostridium perfringens was used to study the intracellular formation of enterotoxin by this organism. Enterotoxin was detected at 4 h of growth at the end of the cell containing forespore. With the development of the spore, enterotoxin accumulation continued and involved the entire length of the cell until its lysis with the release of enterotoxin and mature spore. The spores did not contain demonstrable enterotoxin. Only a certain number of the sporulated cells of the enterotoxigenic strains studied produced this toxin. The amount of enterotoxin produced varied with sporulation percentage, and between strains and individual cells.

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Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2005000100002 ◽

2005 ◽

Vol 25 (1) ◽

pp. 4-8 ◽

Cited By ~ 14

Author(s):

Ronnie A. Assis ◽

Francisco C.F. Lobato ◽

Rogéria Serakides ◽

Renato L. Santos ◽

Guilherme R.C. Dias ◽

...

Keyword(s):

Guinea Pigs ◽

Fluorescent Antibody ◽

Deae Cellulose ◽

Fluorescein Isothiocyanate ◽

Type A ◽

Fluorescent Antibody Technique ◽

Antibody Technique ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Formalin Fixed

Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

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Experimental Food Poisoning with Heat-Susceptible Clostridium perfringens Type A

Journal of Food Science ◽

10.1111/j.1365-2621.1967.tb09713.x ◽

1967 ◽

Vol 32 (4) ◽

pp. 467-469 ◽

Cited By ~ 20

Author(s):

A. H. W. HAUSCHILD ◽

F. S. THATCHER

Keyword(s):

Clostridium Perfringens ◽

Food Poisoning ◽

Type A

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Ulcerative Enteritis-Like Disease Associated With Clostridium Perfringens Type A In Bobwhite Quail (Colinus virginianus)

Avian Diseases Digest ◽

10.1637/1933-5334-3.4.e23 ◽

2008 ◽

Vol 3 (4) ◽

pp. e23-e24

Author(s):

H. L. Shivaprasad ◽

Francisco Uzal ◽

Randy Kokka ◽

Derek J. Fisher ◽

Bruce A. Mcclane ◽

...

Keyword(s):

Clostridium Perfringens ◽

Colinus Virginianus ◽

Type A ◽

Bobwhite Quail

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Immunobiology of Clostridium perfringens Type A: Passive Protection of NMRI Mice

Chemotherapy ◽

10.1159/000238874 ◽

1991 ◽

Vol 37 (5) ◽

pp. 318-326

Author(s):

Walter H. Traub ◽

Dierk Bauer ◽

Ursula Wolf

Keyword(s):

Clostridium Perfringens ◽

Type A ◽

Passive Protection ◽

Nmri Mice

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Adhesive properties of an outer structure of Clostridium perfringens type A isolated from piglets with with catarrhal enteritis

Revista de Microbiologia ◽

10.1590/s0001-37141999000300010 ◽

1999 ◽

Vol 30 (3) ◽

pp. 242-248 ◽

Cited By ~ 2

Author(s):

Elizabeth Pelosi Teixeira ◽

Marlene Braide Serafim ◽

Maria Alice Cruz Höfling ◽

Aureo T. Yamada ◽

Antonio Fernando Pestana de Castro

Keyword(s):

Clostridium Perfringens ◽

Type A ◽

Negative Control ◽

Bacterial Cells ◽

Intestinal Epithelial ◽

Adhesive Properties ◽

Ileal Loop ◽

Fibrillar Material ◽

Gram Staining ◽

Transmission Electron

One strain (S32) of Clostridium perfringens type A was isolated from a case of catarrhal enteritis of piglets. This strain was able to adhere to HeLa cells showing an adherence index (AI) of 25.15 ± 1.26 (mean ± 1 standard error of the mean). Treatment of the bacterial cells with trypsin (0.25mg/ml) decreased in 70%-80% the AI and metaperiodate (10mg/ml) abolished completely the adherence, suggesting that the structure responsible for this phenomenon was probably a glycoprotein. Heating of bacterial suspensions (100ºC/5 min) before carrying out the adhesion test decreased the AI rendering it equal to the negative controls. Rabbit homologous S32 antiserum inhibited the adherence up to dilutions of 1: 640, at least. The piglet ileal loop assay, carried out with strains S32 and Jab-1 (negative control) demonstrated that the strain S32 was able to adhere to the intestinal epithelial cells when examined after Gram staining. Transmission electron microcopy (TEM) demonstrated that S32 strain displayed a loose fibrillar material not seen with Jab-1. Stabilization of the bacterial cells with homologous antiserum of strain S32, followed by staining with rhuteniun red, revealed loose long fibrillar material on the outer surface of the cells, that sometimes could be seen spreading out from the cells and linking bacterial cells. The question whether this structure might be an adhesin for this strain of Cl. perfringes type A, perhaps playing a role in the pathogenesis of the catarrhal enteritis of piglets, is dependent on further studies.

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