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Background  Tick-borne diseases (TBDs) are very important in relation to domestic ruminants, but their occurrence among wild ruminants, mainly in the African buffalo  Syncerus caffer , remains little known.    Methods  Molecular diagnostic methods were applied to detect  Anaplasma marginale ,  Anaplasma centrale ,  Anaplasma phagocytophilum, Ehrlichia ruminantium  and  Ehrlichia chaffeensis  in 97 blood samples of African buffalo captured at the Marromeu Reserve in Mozambique. Molecular detection of agents belonging to the family Anaplasmataceae were based on conventional and qPCR assays based on  msp5 ,  groEL ,  16S  rRNA,  msp2 ,  pCS20  and  vlpt  genes. Phylogenetic reconstruction of new  Anaplasma  isolates detected in African buffalo was evaluated based on  msp5 ,  groEL  and  16S  rRNA genes.    Results  All the animals evaluated were negative for specific PCR assays for  A. phagocytophilum ,  E. ruminantium  and  E. chaffeensis,  but 70 animals were positive for  A. marginale,  showing 2.69 × 10 0  up to 2.00 × 10 5  msp1β  copies/μl. This result overcomes the conventional PCR for  A. marginale  based on  msp5  gene that detected only 65 positive samples. Sequencing and phylogenetic analyses were performed for selected positive samples based on the genes  msp5 ,  groEL  and  16S  rRNA. Trees inferred using different methods separated the 29  msp5  sequences from buffalo in two distinct groups, assigned to  A. centrale  and  A. marginale . The  groEL  sequences determined for African buffalo samples revealed to be more heterogeneous and inferred trees could not assign them to any species of  Anaplasma  despite being more related to  A. marginale  and  A. centrale . The highly conserved  16S  rRNA gene sequences suggested a close relationship of the new 16 sequences with  A. centrale / A. marginale ,  A. platys  and  A. phagocytophilum .    Conclusions  Our analysis suggests that different species of  Anaplasma  are simultaneously present in the African buffalo. To the best of our knowledge, this is the first study that diagnosed  Anaplasma  spp. in the African buffalo and inferred the taxonomic status of new isolates with different gene sequences. The small fragment of  msp5  sequences revealed to be a good target for phylogenetic positioning of new  Anaplasma  spp. isolates.

parasites and vectors

Molecular diagnosis and genetic diversity of tick-borne Anaplasmataceae agents infecting the African buffalo Syncerus caffer from Marromeu Reserve in Mozambique