ABSTRACT
Clostridium perfringens possesses at least two functional quorum sensing (QS) systems, i.e. an Agr-like system and a LuxS-dependent AI-2 system. Both of those QS systems can reportedly control in vitro toxin production by C. perfringens but their importance for virulence has not been evaluated. Therefore, the current study assessed whether these QS systems might regulate the pathogenicity of CN3685, a C. perfringens type C strain. Since type C isolates cause both haemorrhagic necrotic enteritis and fatal enterotoxemias (where toxins produced in the intestines are absorbed into the circulation to target other internal organs), the ability of isogenic agrB or luxS mutants to cause necrotizing enteritis in rabbit small intestinal loops or enterotoxemic lethality in mice was evaluated. Results obtained strongly suggest that the Agr-like QS system, but not the LuxS-dependent AI-2 QS system, is required for CN3685 to cause haemorrhagic necrotizing enteritis, apparently because the Agr-like system regulates the production of beta toxin, which is essential for causing this pathology. The Agr-like system, but not the LuxS-mediated AI-2 system, was also important for CN3685 to cause fatal enterotoxemia. These results provide the first direct evidence supporting a role for any QS system in clostridial infections.
Subject(s)
Bacterial Toxins/biosynthesis , Clostridium Infections/microbiology , Clostridium perfringens/metabolism , Clostridium perfringens/pathogenicity , Quorum Sensing , Animals , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Bacterial Toxins/genetics , Cell Line , Clostridium perfringens/genetics , Clostridium perfringens/isolation & purification , Female , Gene Expression Regulation, Bacterial , Humans , Male , Mice , RabbitsABSTRACT
Clostridium perfringens type D isolates cause enterotoxemia in sheep, goats, and probably cattle. While the major disease signs and lesions of type D animal disease are usually attributed to epsilon toxin, a class B select agent, these bacteria typically produce several lethal toxins. Understanding of disease pathogenesis and development of improved vaccines are hindered by the lack of a small-animal model mimicking natural disease caused by type D isolates. Addressing this need, we developed an oral challenge mouse model of C. perfringens type D enterotoxemia. When BALB/c mice with a sealed anus were inoculated by intragastric gavage with type D isolates, 7 of 10 type D isolates were lethal, as defined by spontaneous death or severe clinical signs necessitating euthanasia. The lethalities of the seven type D isolates varied between 14 and 100%. Clinical signs in the lethally challenged mice included seizures, convulsions, hyperexcitability, and/or depression. Mild intestinal gas distention and brain edema were observed at necropsy in a few mice, while histology showed multifocal acute tubular necrosis of the kidney and edema in the lungs of most challenged mice that developed a clinical response. When the lethality of type D isolates in this model was compared with in vitro toxin production, only a limited correlation was observed. However, mice could be protected against lethality by intravenous passive immunization with an epsilon toxin antibody prior to oral challenge. This study provides an economical new model for studying the pathogenesis of C. perfringens type D infections.
Subject(s)
Clostridium Infections/microbiology , Clostridium perfringens/pathogenicity , Disease Models, Animal , Enterotoxemia/microbiology , Administration, Oral , Animals , Antibodies, Bacterial/administration & dosage , Antibodies, Bacterial/therapeutic use , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/therapeutic use , Bacterial Toxins/biosynthesis , Clostridium Infections/immunology , Clostridium Infections/mortality , Clostridium perfringens/immunology , Clostridium perfringens/isolation & purification , Duodenum/metabolism , Duodenum/microbiology , Enterotoxemia/metabolism , Enterotoxemia/mortality , Immunization, Passive , Intubation, Gastrointestinal , Mice , Mice, Inbred BALB CABSTRACT
A goat kid died after being depressed for several days. No significant gross abnormalities were observed at postmortem examination, while histopathological analysis revealed diffuse necrotizing enterocolitis. Isolation of Clostridium perfringens type A secreting enterotoxin (CPE) and presence of CPE in the small intestine suggest that CPE contributed to the death of this kid.