Clostridium Septicum Research Articles

Sudden Death of a Bearded Vulture (Gypaetus barbatus) Possibly Caused by Newcastle Disease Virus

An adult female bearded vulture (Gypaetus barbatus) in the Tel Aviv University Research Zoo was found dead without previous clinical signs. The predominant pathologic changes were considerable bloody content in the intestines and enlargement of the liver, which had a rubbery consistency with color changes. Microscopic lesions consisted of multifocal histiocytic infiltration in the liver. Newcastle disease virus (NDV) was isolated from a cloacal swab and from the lungs and liver. Intracerebral pathogenicity index of the virus, as estimated in 1-day-old chicks, was repeated three times and had an average value of 1.68, indicating a velogenic strain. Numerous Clostridium septicum bacteria were found on the intestinal surface, but bioassays in which they were orally administered into chickens and mice revealed that, even though they were heavily multiplied in the intestines, they were nonpathogenic. It seems that NDV, documented for the first time in a bearded vulture in Israel, was the likely cause of sudden death.

Clinical features of clostridial bacteremia: a review from a rural area.

Blood samples, which were obtained from patients who lived in a rural area with approximately 500 acute-care hospital beds, were cultured from 1990 through 1997. We retrospectively reviewed the blood cultures that yielded Clostridium species (74 [0.12%] of 63,296 cultures). These were obtained from 46 different hospitalized patients (incidents per hospital, 0.03%). The source of the Clostridium species was a gastrointestinal site in 24 patients (52.2%). The most frequently identified Clostridium species was Clostridium perfringens (in 10 [21.7%] of patients), followed by Clostridium septicum (in 9 [19.6%]). Thirty-one patients (67.4%) were aged > or =65 years, 13 patients (28.3%) had diabetes mellitus, and underlying malignancy was present in 22 patients (47.8%). The mortality rate of patients whose condition had been managed surgically was 33%; for those patients whose conditions required medical management, the mortality rate was 58%. Clostridium bacteremia in these patients usually had a gastrointestinal source, it often occurred in patients with serious underlying medical conditions, and it rarely was the result of traumatic farm accidents.

Isolement de <em>Clostridium septicum</em> à l’occasion d’une épizootie de charbon symptomatique à Madagascar

Des foyers de charbon symptomatique ont fait leur apparition dans différentes localités de la région de Vakinankaratra (Madagascar) au cours des années 1996 et 1997 et ont causé la mort de 588 bovins en l’espace de sept mois. Même les animaux qui avaient été vaccinés entre avril et août 1996 n’ont pas été épargnés. Les analyses bactériologiques entreprises sur les prélèvements reçus ont permis de mettre en évidence en mars 1997 un autre agent du charbon symptomatique, Clostridium septicum, dont l’existence n’avait pas auparavant été rapportée dans le pays. Le diagnostic a été confirmé en mai par l’Institut pour la biotechnologie appliquée aux pays tropicaux de Goettingen (Allemagne) au moyen de la technique de la chromatographie en phase gazeuse. Un antigène vaccinal expérimental a été élaboré à partir du germe pour combler la valence manquante dans le produit conventionnel. L’isolement de cette bactérie survient 28 ans après les premières flambées de charbon symptomatique dues à Clostridium chauvoei. Son origine est discutée brièvement par les auteurs.

Rapid detection and identification of Clostridium chauvoei by PCR based on flagellin gene sequence

We developed a one-step polymerase chain reaction (PCR) system that specifically detects Clostridium chauvoei. Oligonucleotide primers were designed to amplify a 516-bp fragment of the structural flagellin gene. The specificity of the PCR was investigated by analyzing 59 strains of clostridia, and seven strain of other genera. A 516-bp fragment could be amplified from all the C. chauvoei strains tested, and no amplification was observed by using DNAs from the other strains tested, including Clostridium septicum. Similarly, this PCR-based method specifically detected C. chauvoei DNA sequences in samples of muscle and exudate of obtained from mice within 12 h of inoculation. In tests using samples of muscle or liver, the limit of detection was about 200 organisms per reaction. These results suggest that the one-step PCR system may be useful for direct detection and identification of C. chauvoei in clinical specimens.

SPONTANEOUS BIFOCAL CLOSTRIDIUM SEPTICUM GAS GANGRENE

Clostridium septicum gas gangrene (myonecrosis) is an acutely painful and rapidly fatal infection occurring in the absence of trauma. Urgent surgery is essential both to control pain and to ensure survival. Most patients who develop this infection have an underlying malignancy and clinicians should be aware of this association. We present a case of bifocal myonecrosis which to our knowledge has not been reported previously.

The Role of Clostridium septicum in Paraneoplastic Sepsis

Abstract Context.—Clostridium septicum infections are rare but often associated with serious if not fatal outcomes. Clostridium septicum infection does not appear to be associated with a single specific defect in cellular or humoral immunity. It has been associated with multiple medical problems, including but not limited to leukemia, malignancy of the bowel, other solid tumors, cyclic neutropenia with enterocolitis, diabetes mellitus, and severe arteriosclerosis. Most cases of C septicum are associated with malignancy, and mortality approaches 100% if care is not rendered within 12 to 24 hours. Objectives.—To evaluate outcomes of patients with C septicum bacteremia, whether treated medically or surgically or both, and to note associated conditions. Design.—Retrospective evaluation of patients found to have C septicum bacteremia in the past 6 years. Setting.—Two teaching hospitals, Brooke Army Medical Center (250 beds) and Wilford Hall Medical Center (292 beds), were the source of our patients. Patients.—...

Clostridium septicum Alpha Toxin Uses Glycosylphosphatidylinositol-anchored Protein Receptors

The alpha toxin produced by Clostridium septicum is a channel-forming protein that is an important contributor to the virulence of the organism. Chinese hamster ovary (CHO) cells are sensitive to low concentrations of the toxin, indicating that they contain toxin receptors. Using retroviral mutagenesis, a mutant CHO line (BAG15) was generated that is resistant to alpha toxin. FACS analysis showed that the mutant cells have lost the ability to bind the toxin, indicating that they lack an alpha toxin receptor. The mutant cells are also resistant to aerolysin, a channel-forming protein secreted by Aeromonas spp., which is structurally and functionally related to alpha toxin and which is known to bind to glycosylphosphatidylinositol (GPI)-anchored proteins, such as Thy-1. We obtained evidence that the BAG15 cells lack N-acetylglucosaminyl-phosphatidylinositol deacetylase-L, needed for the second step in GPI anchor biosynthesis. Several lymphocyte cell lines lacking GPI-anchored proteins were also shown to be less sensitive to alpha toxin. On the other hand, the sensitivity of CHO cells to alpha toxin was increased when the cells were transfected with the GPI-anchored folate receptor. We conclude that alpha toxin, like aerolysin, binds to GPI-anchored protein receptors. Evidence is also presented that the two toxins bind to different subsets of GPI-anchored proteins.

Detection of neutralizing antibodies against alpha-toxin of different Clostridium septicum strains in cell culture.

Clostridium septicum, a ubiquitious organism, is the pathogen which causes the classical malignant edema after injuries. Because of its strong cytotoxic alpha-toxin, infections are often lethal. To prevent losses in animals, vaccination with alpha-toxoid vaccines is carried out. Quality control of the vaccines is done by a neutralization test in mice. A cytotoxin test and as an alternative method to detect neutralizing antibodies, a cytotoxin inhibition test was standardized. In the studies, alpha-toxin of the C. septicum reference strain (NC 547) from the National Collection of Type Cultures was compared with alpha-toxin of a field strain from an outbreak in Germany. Sera from five heterologous polyvalent and three monovalent vaccines from eight rabbit groups were available. Each vaccination had been carried out according to the procedure of the German Pharmacopoeia. In three out of the five sera of the groups vaccinated with the heterologous polyvalent vaccine, cytotoxin neutralizing antibodies were detected. High antibody titers were observed in sera of rabbits vaccinated with a vaccine of strain NC 547, lower titers in the sera of rabbits vaccinated with a vaccine of the field strain. No cytotoxin neutralizing antibodies could be found in the sera of rabbits vaccinated with the monovalent C. chauvoei vaccine. The toxins of all strains showed the same ranking of the vaccines. Vaccines which caused high antibody titers in the animals were detected by all toxins as such, as well as vaccines which had medium or low antibody inducing capacity. The results were independent of the C. septicum strain used for the production of alpha-toxin.

Spontaneous Myonecrosis Secondary to Clostridium septicum

Spontaneous Myonecrosis Secondary to Clostridium septicum

Detection of neutralizing antibodies against α-toxin of different Clostridium septicum strains in cell culture

Clostridium septicum, a ubiquitious organism, is the pathogen which causes the classical malignant edema after injuries. Because of its strong cytotoxic α-toxin, infections are often lethal. To prevent losses in animals, vaccination with α-toxoid vaccines is carried out. Quality control of the vaccines is done by a neutralization test in mice. A cytotoxin test and as an alternative method to detect neutralizing antibodies, a cytotoxin inhibition test was standardized. In the studies, α-toxin of the C. septicum reference strain (NC 547) from the National Collection of Type Cultures was compared with α-toxin of a field strain from an outbreak in Germany. Sera from five heterologous polyvalent and three monovalent vaccines from eight rabbit groups were available. Each vaccination had been carried out according to the procedure of the German Pharmacopoeia. In three out of the five sera of the groups vaccinated with the heterologous polyvalent vaccine, cytotoxin neutralizing antibodies were detected. High antibody titers were observed in sera of rabbits vaccinated with a vaccine of strain NC 547, lower titers in the sera of rabbits vaccinated with a vaccine of the field strain. No cytotoxin neutralizing antibodies could be found in the sera of rabbits vaccinated with the monovalent C. chauvoei vaccine. The toxins of all strains showed the same ranking of the vaccines. Vaccines which caused high antibody titers in the animals were detected by all toxins as such, as well as vaccines which had medium or low antibody inducing capacity. The results were independent of the C. septicum strain used for the production of α-toxin.

Characterization of 1.8 kb Hind III Fragment of Clostridium septicum Genomic DNA

Narayanan, S., Singh, S.D. and Bakshi, C.S. 1999. Characterization of 1.8 kb Hind III fragment of Clostridium septicum genomic DNA. J. Appl. Anim. Res., 15: 185–190. A genomic library of Clostridium septicum (IVRI strain 51) was generated by cloning Hind III fragments of genomic DNA of C. septicum into the plasmid pUC18. Thirty-three recombinant clones were screened and a recombinant, plasmid (pSDS12) containing a 1.8 kb insert was selected for characterization by restriction mapping. The map was confirmed by probing the restriction digests of the insert with 1.8 kb insert labeled with Digoxigenin-d-UTP. This insert has a unique Nsi 1 site. This Nsi 1 site could facilitate further sub-cloning as preliminary studies indicate that this insert may show utility as a gene probe in the diagnosis of C. septicum infections.

Clostridium septicum osteomyelitis with intramedullary gas on CT and MR imaging.

Clostridium septicum osteomyelitis with intramedullary gas on CT and MR imaging.

Wound infection with Clostridium septicum after intramuscular injection in a pony

Wound infection with Clostridium septicum after intramuscular injection in a pony

Clostridium septicum Infection in a Patient with Felty??s Syndrome

From the Department of Internal Medicine, Salt Lake Citv VA Medical Center, University of Utah Medical Center, Salt Lake Citv, Utah

Early Clostridium septicum endocarditis and myocarditis after reconstruction of congenital heart disease

Early Clostridium septicum endocarditis and myocarditis after reconstruction of congenital heart disease

Effect of temperature and glucose concentration on DNase production by Clostridium septicum.

Effect of temperature and glucose concentration on DNase production by Clostridium septicum.

Another cause of necrotizing fasciitis?

Necrotizing fasciitis is a rare condition, but has a devastating clinical course if it is not diagnosed early and treated aggressively. There are usually a number of organisms implicated in its pathogenesis, but in this case report a patient is presented in whom the only organism isolated from both tissue and blood cultures was Clostridium septicum. This organism almost always causes a myonecrosis, but in this case only the superficial and deep fascial layers were affected, sparing the underlying muscle. Clostridium septicum resulting in a true necrotizing fasciitis with no myonecrosis and being the only isolate from both blood and tissue cultures has not been previously reported in literature.

Antibody responses to clostridial infection in humans.

Serum antibody responses to the major toxins produced by Clostridium difficile, Clostridium perfringens, Clostridium septicum, Clostridium tetani, and Clostridium botulinum have been documented following infection. Effective toxoid vaccines for tetanus and enteritis necroticans due to C. perfringens type C demonstrate the potential of antitoxin responses. Although individual serum and mucosal antibody responses to C. difficile enterotoxin (toxin A) vary, one-third of patients with C. difficile diarrhea develop neutralizing serum antibodies. IgA in the serum, not IgG, is typically responsible for this neutralization, suggesting a unique role for serum IgA in response to C. difficile infection, an infection that is usually limited to the intestinal mucosa. The relationship of naturally occurring antitoxin antibodies to the clinical course of C. difficile infection is controversial. However, patients with chronic relapsing C. difficile diarrhea and low levels of IgG to toxin A have shown clinical responses following intravenous therapy with immune globulin. Antibody responses to non-toxin C. difficile proteins also occur, but their significance is only partially known.

Identification of Clostridium chauvoei in cultures and clinical material from blackleg using PCR

An identification system for Clostridium chauvoei, using PCR amplification of the 16S rRNA gene ( rrs) with specific oligonucleotide primers and subsequent restriction digestion of the amplification product is described. The specific oligonucleotide primers were designed based on the rrs gene sequences of C. chauvoei by comparing it to the DNA sequences of the rrs genes of its most closely related species Clostridium septicum and Clostridium carnis. A subsequent restriction digestion of the 960 bp amplification product was used in order to unambiguously identify C. chauvoei. The developed identification system was evaluated on clinical material during a recent outbreak of blackleg in cattle. Thereby, C. chauvoei was identified as the etiologic agent of the outbreak either directly from clinical samples of muscle, liver, spleen and kidney or from primary cultures made with this material. A comparison of the newly developed method with standard diagnostic tools for C. chauvoei showed that it has advantages over the immunofluorescence and is, therefore, a useful option to it. Moreover, the assay is a valuable tool for the phylogenetic identification of C. chauvoei which can assist to substitute the fastidious traditional identification methods and replace laboratory animal testing currently used.

The propeptide of Clostridium septicum alpha toxin functions as an intramolecular chaperone and is a potent inhibitor of alpha toxin-dependent cytolysis.

Clostridium septicum alpha toxin is activated by a proteolytic cleavage at Arg-398 in its carboxy terminus, which yields a 41.3-kDa cytolytically active toxin and a 5.1-kDa propeptide. Studies were performed to determine when the propeptide dissociated from the toxin after proteolytic activation of the protoxin (AT(pro)) and to demonstrate the chaperone activity of the propeptide. The propeptide was found to remain associated with the toxin after activation with trypsin (AT(act)) when analysed by gel filtration or affinity chromatography of a polyhistidine-tagged derivative that contained the polyhistidine tag on the propeptide. The affinity of the propeptide for the toxin was decreased significantly when a mutation was introduced in which Val-400 was converted to a cysteine residue. This mutation destabilized the interaction of the propeptide with the toxin and the propeptide was found to dissociate from the toxin under the same gel-filtration conditions used for the wild-type toxin. The separation of the propeptide in the V400C mutant did not affect the cytolytic activity of the toxin and therefore the propeptide was not necessary for cytolytic activity. These data suggested that the propeptide did not dissociate from the main body of the toxin after proteolysis. Further analysis demonstrated that purified propeptide was a potent inhibitor of alpha toxin activity, which inhibited the oligomerization of alpha toxin into a functional pore. These data suggest that the propeptide does not participate in the final oligomerized complex and that oligomerization appears to displace the propeptide from AT(act). The importance of the propeptide to the solution stability of alpha toxin was also demonstrated. When AT(pro) was activated in solution with trypsin a significant level (approximately 50%) of inactive aggregate formed. This aggregate, which could be removed by centrifugation at 14,000 x g, was made up of both SDS-sensitive and -resistant aggregates, suggesting that a variety of inactive aggregates formed when the monomers interacted in solution. Significantly higher levels of haemolytic activity (approximately 16-fold) were observed when alpha toxin was proteolytically activated after membrane binding instead of in solution. These results support the role of the propeptide as an intramolecular chaperone that stabilizes the monomeric AT(pro) and shuttles it to the membrane where it is activated by protease, oligomerizes into a pre-pore complex and forms a pore. The data suggest that oligomerization of the toxin displaces the propeptide from the monomer form of alpha toxin and that the propeptide does not participate in, and is not necessary to, the final cytolytic complex.