The effect of Clostridium perfringens type C strain CN3685 and its isogenic beta toxin null mutant in goats

Abstract

Clostridium perfringens type C is an important cause of enteritis and/or enterocolitis in several animal species, including pigs, sheep, goats, horses and humans. The disease is a classic enterotoxemia and the enteric lesions and associated systemic effects are thought to be caused primarily by beta toxin (CPB), one of two typing toxins produced by C. perfringens type C. This has been demonstrated recently by fulfilling molecular Koch's postulates in rabbits and mice. We present here an experimental study to fulfill these postulates in goats, a natural host of C. perfringens type C disease. Nine healthy male or female Anglo Nubian goat kids were inoculated with the virulent C. perfringens type C wild-type strain CN3685, an isogenic CPB null mutant or a strain where the cpb null mutation had been reversed. Three goats inoculated with the wild-type strain presented abdominal pain, hemorrhagic diarrhea, necrotizing enterocolitis, pulmonary edema, hydropericardium and death within 24h of inoculation. Two goats inoculated with the CPB null mutant and two goats inoculated with sterile culture media (negative controls) remained clinically healthy during 24h after inoculation and no gross or histological abnormalities were observed in the tissues of any of them. Reversal of the null mutation to partially restore CPB production also increased virulence; 2 goats inoculated with this reversed mutant presented clinical and pathological changes similar to those observed in goats inoculated with the wild-type strain, except that spontaneous death was not observed. These results indicate that CPB is required for C. perfringens type C to induce disease in goats, supporting a key role for this toxin in natural C. perfringens type C disease pathogenesis.

Fig. 1

Jejunum of a goat inoculated with C. perfringens type C wild-type. Observe the presence of a pseudomembrane and blood in the intestinal lumen.

Fig. 2

Jejunum of a goat inoculated with C. perfringens type C CPB null mutant. No gross lesions are observed.

Fig. 3

Jejunum of a goat inoculated with C. perfringens type C CPB reverse mutant. The mucosa is hemorrhagic and there is fibrin present in the intestinal lumen.

Fig. 4

Jejunum of a goat inoculated with sterile, non-toxic TGY. No gross lesions are observed.

Fig. 5

Jejunum of a goat inoculated with C. perfringens type C wild type. The mucosa and submucosa are diffusely necrotic. Bar: 100µm

Fig. 6

Jejunum of a goat inoculated with C. perfringens type C CPB null mutant. No significant histological lesions are observed. Bar: 100µm

Fig. 7

Jejunum of a goat inoculated with C. perfringens type C CPB reverse mutant. There is diffuse mucosal necrosis. Bar: 100µm

Fig. 8

Jejunum of a goat inoculated with TGY. No significant histological lesions are observed. Bar: 100µm

Fig. 9

Higher magnification of the goat’s jejunum showed in Figure 5. There is complete loss of villus and crypt epithelium and necrosis of the lamina propria. Bar: 40 µm

Fig. 10

Higher magnification of the goat’s jejunum showed in Figure 6. No significant histological abnormalities are observed. Bar: 40 µm

Fig. 11

Higher magnification of the goat’s jejunum showed in Figure 7. There is partial loss of villus epithelium. Bar: 40 µm

Fig. 12

Higher magnification of the goat’s jejunum showed in Figure 8. No significant histological abnormalities are observed. Bar: 40 µm