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Anaplasmataceae agents are obligatory intracellular Gram-negative α-proteobacteria that are transmitted mostly by arthropod vectors. Although mammals of the Superorder Xenarthra (sloths, anteaters, and armadillos) have been implicated as reservoirs for several zoonotic agents, only few studies have sought to detect Anaplasmataceae agents in this group of mammals. This study aimed to investigate the occurrence and genetic diversity of  Anaplasma  spp. and  Ehrlichia  spp. in blood and spleen samples of free-living Xenarthra from four different states in Brazil (São Paulo, Mato Grosso do Sul, Rondônia, and Pará). Nested and conventional PCR screening assays were performed to detect the  rrs  and  dsb  genes of  Anaplasma  spp. and  Ehrlichia  spp., respectively. The assays were positive in 27.57% (91/330) of the  Anaplasma  spp. and 24.54% (81/330) of the  Ehrlichia  spp. Of the 91 positive  Anaplasma  spp. samples, 56.04% were positive in a conventional PCR assay targeting the 23S–5S intergenic region. Phylogenetic and distance analyses based on the  rrs  gene allocated  Anaplasma  sequences from sloths captured in Rondônia and Pará states in a single clade, which was closely related to the  A. marginale ,  A. ovis,  and  A. capra  clades. The sequences detected in southern anteaters from São Paulo were allocated in a clade closely related to sequences of  Anaplasma  spp. detected in  Nasua nasua ,  Leopardus pardalis , and  Cerdocyon thous  in Brazil. These sequences were positioned close to  A. odocoilei  sequences. Genotype analysis corroborated previous findings and demonstrated the circulation of two distinct  Anaplasma  genotypes in animals from north and southeast Brazil. The first genotype was new. The second was previously detected in  N. nasua  in Mato Grosso do Sul state. The intergenic region analyses also demonstrated two distinct genotypes of  Anaplasma . The sequences detected in Xenarthra from Pará and Rondônia states were closely related to those in  A. marginale ,  A. ovis,  and  A. capra .  Anaplasma  spp. sequences detected in Xenarthra from São Paulo and were allocated close to those in  A. phagocytophilum . The analyses based on the  dsb  gene grouped the  Ehrlichia  spp. sequences with sequences of  E. canis  (São Paulo, Mato Grosso do Sul, and Pará) and  E. minasensis  (Rondônia and Pará). The data indicate the occurrence of  E. canis  and  E. minasensis  and two possible new  Candidatus  species of  Anaplasma  spp. in free-living mammals of the Superorder Xenarthra in Brazil.

Ehrlichia spp. and Anaplasma spp. in Xenarthra mammals from Brazil, with evidence of novel ‘Candidatus Anaplasma spp.’