Use of Genetically Manipulated Strains of Clostridium perfringens Reveals that Both Alpha-Toxin and Theta-Toxin Are Required for Vascular Leukostasis To Occur in Experimental Gas Gangrene

ABSTRACT A hallmark of gas gangrene (clostridial myonecrosis) pathology is a paucity of leukocytes infiltrating the necrotic tissue. The cause of this paucity most likely relates to the observation of leukocyte aggregates at the border of the area of tissue necrosis, often within the microvasculature itself. Infecting mice with genetically manipulated strains of Clostridium perfringens type A (deficient in either alpha-toxin or theta-toxin production) resulted in significantly reduced leukocyte aggregation when alpha-toxin was absent and complete abrogation of leukocyte aggregation when theta-toxin was absent. Thus, both alpha-toxin and theta-toxin are necessary for the characteristic vascular leukostasis observed in clostridial myonecrosis.

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Perfringolysin O Expression in Clostridium perfringens Is Independent of the Upstream pfoR Gene

Journal of Bacteriology ◽

10.1128/jb.184.7.2034-2038.2002 ◽

2002 ◽

Vol 184 (7) ◽

pp. 2034-2038 ◽

Cited By ~ 5

Author(s):

Milena M. Awad ◽

Julian I. Rood

Keyword(s):

Escherichia Coli ◽

Homologous Recombination ◽

Gene Product ◽

Structural Gene ◽

Clostridium Perfringens ◽

Deletion Mutant ◽

Gas Gangrene ◽

Alpha Toxin ◽

Clostridial Myonecrosis ◽

Perfringolysin O

ABSTRACT The pathogenesis of Clostridium perfringens-mediated gas gangrene or clostridial myonecrosis involves the extracellular toxins alpha-toxin and perfringolysin O. Previous studies (T. Shimizu, A. Okabe, J. Minami, and H. Hayashi, Infect. Immun. 59:137-142, 1991) carried out with Escherichia coli suggested that the perfringolysin O structural gene, pfoA, was positively regulated by the product of the upstream pfoR gene. In an attempt to confirm this hypothesis in C. perfringens, a pfoR-pfoA deletion mutant was complemented with isogenic pfoA+ shuttle plasmids that varied only in their ability to encode an intact pfoR gene. No difference in the ability to produce perfringolysin O was observed for C. perfringens strains carrying these plasmids. In addition, chromosomal pfoR mutants were constructed by homologous recombination in C. perfringens. Again no difference in perfringolysin O activity was observed. Since it was not possible to alter perfringolysin O expression by mutation of pfoR, it was concluded that the pfoR gene product is unlikely to have a role in the regulation of pfoA expression in C. perfringens.

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Synergistic Effects of Alpha-Toxin and Perfringolysin O in Clostridium perfringens-Mediated Gas Gangrene

Infection and Immunity ◽

10.1128/iai.69.12.7904-7910.2001 ◽

2001 ◽

Vol 69 (12) ◽

pp. 7904-7910 ◽

Cited By ~ 119

Author(s):

Milena M. Awad ◽

Darren M. Ellemor ◽

Richard L. Boyd ◽

John J. Emmins ◽

Julian I. Rood

Keyword(s):

Structural Gene ◽

Clostridium Perfringens ◽

Synergistic Effects ◽

Gas Gangrene ◽

Alpha Toxin ◽

Coagulative Necrosis ◽

Clostridial Myonecrosis ◽

Perfringolysin O ◽

Vascular Disruption ◽

Rapid Spread

ABSTRACT To examine the synergistic effects of alpha-toxin and perfringolysin O in clostridial myonecrosis, homologous recombination was used to construct an alpha-toxin deficient derivative of a perfringolysin O mutant of Clostridium perfringens. The subsequent strain was complemented with separate plasmids that carried the alpha-toxin structural gene (plc), the perfringolysin O gene (pfoA), or both toxin genes, and the resultant isogenic strains were examined in a mouse myonecrosis model. Synergistic effects were clearly observed in these experiments. Infection with the control strain, which did not produce either toxin, resulted in very minimal gross pathological changes, whereas the isogenic strain that was reconstituted for both toxins produced a pathology that was clearly more severe than when alpha-toxin alone was reconstituted. These changes were most apparent in the rapid spread of the disease, the gross pathology of the footpad and in the rate at which the mice had to be euthanatized for ethical reasons. Elimination of both alpha-toxin and perfringolysin O production removed most of the histopathological features typical of clostridial myonecrosis. These effects were restored when the mutant was complemented with the alpha-toxin structural gene, but reconstituting only perfringolysin O activity produced vastly different results, with regions of coagulative necrosis, apparently enhanced by vascular disruption, being observed. Reconstitution of both alpha-toxin and perfringolysin O activity produced histopathology most similar to that observed with the alpha-toxin reconstituted strain. The spreading of myonecrosis was very rapid in these tissues, and coagulative necrosis appeared to be restricted to the lumen of the blood vessels. The results of these virulence experiments clearly support the hypothesis that alpha-toxin and perfringolysin O have a synergistic effect in the pathology of gas gangrene.

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Experimental gas gangrene with food-poisoning Clostridium perfringens type A

Canadian Journal of Microbiology ◽

10.1139/m68-117 ◽

1968 ◽

Vol 14 (6) ◽

pp. 705-709 ◽

Cited By ~ 7

Author(s):

A. H. W. Hauschild ◽

F. S. Thatcher

Keyword(s):

Clostridium Perfringens ◽

Food Poisoning ◽

Type A ◽

Gas Gangrene ◽

Sheep Blood Agar ◽

Alpha Toxin ◽

Cell Numbers ◽

Fatal Infection ◽

Virulent Strains ◽

Resistant Strains

Classical and food-poisoning strains of Clostridium perfringens type A were tested for their capacity to produce gas gangrene in guinea pigs.The virulence of food-poisoning strains producing heat-sensitive spores and showing beta hemolysis on sheep-blood agar was comparable to that of the classical strains. The most virulent strains of both groups produced fatal infection with only three to five vegetative cells. Of 13 food-poisoning, heat-sensitive strains showing no beta hemolysis, only three were lethal when a minimum of 4 × 104 to 4 × 108 cells was injected. None of the food-poisoning, heat-resistant strains produced fatal infection with cell numbers up to 4 × 108. The groups of strains showed a correlation between virulence and formation of alpha toxin in liquid culture.It is concluded that a number of heat-sensitive, beta-hemolytic strains of C. perfringens may cause gas gangrene as well as food poisoning, and that the current subdivision of C. perfringens type A strains into classical and food-poisoning groups is no longer tenable.

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A retrospective study on the diagnosis of clostridial myonecrosis in ruminants in Brazil

Ciência Rural ◽

10.1590/0103-8478cr20160492 ◽

2017 ◽

Vol 47 (1) ◽

Cited By ~ 2

Author(s):

Prhiscylla Sadanã Pires ◽

Roselene Ecco ◽

Rodrigo Otávio Silveira Silva ◽

Marina Rios de Araújo ◽

Felipe Masiero Salvarani ◽

...

Keyword(s):

Retrospective Study ◽

Clostridium Perfringens ◽

Type A ◽

Gas Gangrene ◽

The Other ◽

Post Mortem ◽

Tissue Samples ◽

Clostridial Myonecrosis ◽

Endemic Areas ◽

Clostridium Chauvoei

ABSTRACT: A standardized immunochemistry method for the diagnosis of clostridial myonecrosis was applied to 38 formalized tissue samples from ruminants with clinical and post mortem history suggestive of blackleg or gas gangrene. The diagnosis of clostridial myonecrosis was confirmed in 37 out of 38 (97.4%) samples tested. Clostridium chauvoei and Clostridium perfringens type A were the most common agents found alone, being detected in ten (26.3%) and six (15.8%) samples, respectively. The other cases showed an association of two or three clostridia, with C. perfringens type A detected in 11 (29%) cases. Based on the findings of the present study, polyvalent vaccines against clostridial infections of animals incorporating C. perfringens would be more adequate for preventative purposes in the endemic areas.

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Amentoflavone Attenuates Clostridium perfringens Gas Gangrene by Targeting Alpha-Toxin and Perfringolysin O

Frontiers in Pharmacology ◽

10.3389/fphar.2020.00179 ◽

2020 ◽

Vol 11 ◽

Author(s):

Shui Liu ◽

Xiaofeng Yang ◽

Hong Zhang ◽

Jian Zhang ◽

Yonglin Zhou ◽

...

Keyword(s):

Clostridium Perfringens ◽

Gas Gangrene ◽

Alpha Toxin ◽

Perfringolysin O

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Chemotherapy of Experimental (Murine) Clostridium perfringens Type A Gas Gangrene

Chemotherapy ◽

10.1159/000238611 ◽

1988 ◽

Vol 34 (6) ◽

pp. 472-477 ◽

Cited By ~ 3

Author(s):

Walter H. Traub

Keyword(s):

Clostridium Perfringens ◽

Type A ◽

Gas Gangrene

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A genetically engineered vaccine against the alpha-toxin of Clostridium perfringens protects mice against experimental gas gangrene

Vaccine ◽

10.1016/0264-410x(93)90051-x ◽

1993 ◽

Vol 11 (12) ◽

pp. 1253-1258 ◽

Cited By ~ 111

Author(s):

E.D. Williamson ◽

R.W. Titball

Keyword(s):

Clostridium Perfringens ◽

Gas Gangrene ◽

Genetically Engineered ◽

Alpha Toxin

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Complication of Invasive Molar Pregnancy withClostridium perfringensSepsis

Case Reports in Obstetrics and Gynecology ◽

10.1155/2014/282141 ◽

2014 ◽

Vol 2014 ◽

pp. 1-4

Author(s):

Sanmeet Singh ◽

Kunal Angra ◽

Bonnie Davis ◽

Babak Shokrani

Keyword(s):

Clostridium Perfringens ◽

Complete Resolution ◽

Past Research ◽

Gas Gangrene ◽

Molar Pregnancy ◽

Malignant Diseases ◽

Necrotic Tissue ◽

Associated Symptoms ◽

History Of ◽

Near Term

Clostridium perfringens(CP) is an anaerobic, Gram-positive bacillus associated with malignant diseases and near-term pregnancies. The necrotic tissue that results from these disease processes fuels the proliferation of CP, leading to gas gangrene and subsequently sepsis. Herein, we report a case of a 41-year-old female patient with a history of invasive molar pregnancy that was further complicated with a CP infection. Although past research has shown a link betweenClostridiuminfection and choriocarcinoma (Chern-Horng and Hsieh, 1999), no previous cases of CP infection have been associated with invasive molar pregnancy. We also report complete resolution of the CP sepsis and its associated symptoms following the hysterectomy.

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