BackgroundTick-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.MethodsMolecular 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.ResultsAll 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 × 100 up to 2.00 × 105msp1β 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.ConclusionsOur 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.

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The present study reports the genetic diversity of Anaplasma marginale during anaplasmosis outbreaks in rural properties of the states of Goiás and São Paulo, Brazil. Mortality rates of 3.5% (37/1,050) in calves, 4.7% (45/954) in heifers and 1.1% (25/2,200) in lactating cows were observed in a cattle herd of the municipality of Mambaí, state of Goiás, central-western Brazil. In a cattle herd from the municipality of Lins, state of São Paulo, in southeastern Brazil, none of the animals died, despite presenting clinical signs suggestive of bovine anaplasmosis and exhibiting a drastic decrease in milk production. Thus, blood samples were collected from 100 animals with clinical signs suggestive of bovine anaplasmosis in the municipalities of Mambaí and Lins. Based on the microsatellite structure of the MSP1a of A. marginale, the genotypes E and H were observed in Lins, and the C, D and E genotypes were found in Mambaí. The analysis of the tandem repeat structures of the MSP1a showed nine different strains (τ-10 -15, α-β 2 , α-β 3 -13, α-β 2 192, τ-β-100, α-β 2 -Γ, 193-β-100, 191-13-Γ and 191-13-18) in Lins and two (α-β 3 -Γ and E-F-φ 2 -F 2 ) in Mambaí. Three new tandem repeats of MSP1a (191, 192 and 193) were described. The τ-10-15 and α-β 3 -Γ strains were predominantly associated with the occurrence of clinical anaplasmosis and mortality in calves, heifers and lactating cows.

Tuberculosis is a disease with a great zoonotic potential. It is considered a major obstacle to cattle production and is responsible for severe losses in several production systems. A comparative cervical test (CCT) was performed in 1140 buffaloes from different mesoregions of the state of Pará, Brazil, with the aim of comparing the sensitivity and specificity of CCT with histopathological examination and bacterial culture. Of the animals tested using CCT, 4.65% (53/1140) were positive, 2.98% (34/1140) were inconclusive, and 92.36% (1053/1140) were negative. Among the 168 sacrificed animals, 33 were positive, 18 were inconclusive, and 117 were negative by CCT, and samples from the sacrificed animals were collected for histopathological examination and bacterial culture. A qualitative evaluation of the tuberculin test was performed by comparing the test results with the histopathological and bacteriological results. The latter two tests yielded a prevalence of 4.16%, a sensitivity of 71.43%, and a specificity of 82.61%. Based on these results, we concluded that CCT yielded satisfactory results and can be applied in diagnostic studies in buffaloes. The prevalence rate obtained using three distinct diagnostic methods suggests that Mycobacterium bovis was present in a few animals in the population evaluated.