Serodiagnosis of caseous lymphadenitis (pseudotuberculosis) in sheep

: Random postmortal isolation of Corynebacterium pseudotuberculosis from the sheep used in a research project and the consequently determined serological positivity of these animals indicated the serological examination of the flock from which the sheep came. No symptoms of caseous lymphadenitis were ever observed in the flock in question and no data on the occurrence of this disease were available in the flock history. The sera of 228 head of sheep were used for examinations, while this number of sheep represented half the flock head. Agar-gel immunodiffusion (AID) and neutralization of the toxin C. pseudotuberculosis (NTX) were applied as serological assays. The antigen in both tests was an isolated form of toxin i.e. phospholipase D (PLD). To determine the effect of PLD, the intensity of its synergic hemolytic effect with the equi factor Rhodococcus equi was found out and it was expressed in activity units (AU). Undiluted serum with PLD containing 5,000 AU/ml was examined in AID. Positive precipitation was observed in 79 examined sera, out of which 71 were also positive in NTX. NTX demonstrated the inhibition of synergic hemolytic activity of PLD with the equi factor, which was used to sensibilize sheep erythrocytes in agar medium. PLD with the terminal content of 10 AU/ml was used, as well as twofold dilution series of sera. The first evaluated dilution series was 1:10. In this dilution series, the highest number of sera, 22, had the positive reaction. The highest positivity at a dilution series 1:5,120 was found out in one serum. A total of 71 sera had positive reactions in NTX and those sera were also positive in AID. The determined 34.7% positivity substantiated the importance of serodiagnosis for the intravital diagnosis of caseous lymphadenitis of sheep, particularly of its chronic form. AID and NTX are usable tests for serodiagnosis of this disease.