Molecular evidence of the reservoir competence of water buffalo (Bubalus bubalis) for Anaplasma marginale in Cuba.

Water buffalo (Bubalus bubalis) is a potential reservoir for Anaplasma marginale in livestock ecosystems of tropical countries. However, their participation in the epidemiological process of bovine anaplasmosis in endemic areas remains unclear. In the present study, the reservoir competence of water buffalo for A. marginale was explored by focusing on the analysis of rickettsemia levels in carrier animals, and the genetic characterization of A. marginale strains from cattle and buffalo. Eight groups of cattle and water buffaloes were randomly selected from cohabiting herds in four livestock ecosystems of Cuba, together with two control groups from unrelated cattle and buffalo herds. A total of 180 adult animals (88 water buffalo and 92 cattle) were sampled. Rickettsemia in carrier animals was determined by quantitative real-time PCR. The rickettsemia (parasitemia) levels in cattle were higher than in buffaloes, however the rickettsemia in buffalo may be enough to infect R. microplus ticks. The genetic diversity of A. marginale was assessed by strain characterization and phylogenetic analysis of 27 msp1α gene sequences. The results showed genetic similarity among strains from cattle and water buffalo, suggesting the occurrence of cross-species transmission.

Molecular identification of Anaplasma marginale in two autochthonous South American wild species revealed an identical new genotype and its phylogenetic relationship with those of bovines.

BACKGROUND: Anaplasma marginale is a well-known cattle pathogen of tropical and subtropical world regions. Even though, this obligate intracellular bacterium has been reported in other host species different than bovine, it has never been documented in Myrmecophaga tridactyla (giant anteater) or Hippocamelus antisense (taruca), which are two native endangered species. METHODS: Samples from two sick wild animals: a Myrmecophaga tridactyla (blood) and a Hippocamelus antisense (blood and serum) were studied for the presence of A. marginale DNA through msp5 gene fragment amplification. Further characterization was done through MSP1a tandem repeats analysis and MLST scheme and the genetic relationship among previously characterized A. marginale sequences were studied by applying, eBURST algorithm and AMOVA analysis. RESULTS: Anaplasma marginale DNA was identified in the Myrmecophaga tridactyla and Hippocamelus antisense samples. Through molecular markers, we identified an identical genotype in both animals that was not previously reported in bovine host. The analysis through eBURST and AMOVA revealed no differentiation between the taruca/anteater isolate and the bovine group. CONCLUSIONS: In the present publication we report the identification of A. marginale DNA in a novel ruminant (Hippocamelus antisense) and non-ruminant (Myrmecophaga tridactyla) host species. Genotyping analysis of isolates demonstrated the close relatedness of the new isolate with the circulation population of A. marginale in livestock. Further analysis is needed to understand whether these two hosts contribute to the anaplasmosis epidemiology.