Anaplasma marginale initial bodies of the Norton Zimbabwe strain were disrupted and separated into two membrane fractions banding at 1.15 and 1.22 g/cm3 by sucrose density centrifugation. The membrane fractions

Heartwater, a major constraint to improved livestock production in Zimbabwe, threatens to invade areas which have been previously unaffected. To monitor its spread in Zimbabwe, an immunoblotting diagnostic assay based on the responses of animals to the immunodominant, conserved 32-kDa protein of Cowdria ruminantium was evaluated. In this assay, no false reactions were detected with sera known to be positive and negative, but sera from some cattle, sheep, and goats from heartwater-free areas of Zimbabwe reacted strongly with the 32-kDa protein, suggesting that either these animals had previous exposure to heartwater or they were false positives. To investigate the possibility of previous exposure to heartwater, 11 immunoblot-positive and 6 immunoblot-negative sheep from heartwater-free areas of Zimbabwe were compared regarding their susceptibilities to challenge with C. ruminantium. Prior to challenge, C. ruminantium could not be detected in any sheep by transmission to Amblyomma hebraeum ticks or by the polymerase chain reaction (PCR) conducted with plasma samples. All sheep were equally susceptible to the challenge, and infection was confirmed by brain biopsy, necropsy, PCR, and transmission of C. ruminantium to ticks. Our data suggest that the immunoblotpositive reactions of sera from heartwater-free areas were due not to previous C. ruminantium infection but rather to antigenic cross-reactivity between C. ruminantium and another agent(s), such as Ehrlichia species. In conclusion, the immunodominant 32-kDa protein is not antigenically specific to C. ruminantium and its use in serological diagnosis of heartwater requires reevaluation.

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Cattle immunised by infection with the Florida stock of Anaplasma marginale were protected against subsequent homologous challenge, as demonstrated by complete prevention of microscopically detectable parasitaemia. Identically immunised cattle were partially protected against challenge with the Norton, Zimbabwe stock of A. marginale as determined by the significant prolongation of the prepatent period, a significantly lower peak parasitaemia, and a significantly lower percentage drop in haemoglobin concentration when compared to non-immunised calves challenged identically. The partial protection induced by live Florida stock immunisation demonstrates that while protection-inducing epitopes are shared between the two stocks, induction of complete immunity against a Zimbabwe stock may require alternative presentation of Florida stock epitopes or inclusion of additional Zimbabwe stock epitopes in the immunogen.

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