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....
The superorder Xenarthra consists of sloths, anteaters and armadillos, mammals originated from South America and currently distributed from the south of North America to the south of South America. The present study aimed to investigate the occurrence and genetic diversity of Bartonella spp. in blood and spleen samples from free‐living Xenarthra mammals in the states of São Paulo (SP), Mato Grosso do Sul (MS), Rondônia (RO) and Pará (PA). Based on a quantitative real‐time PCR (qPCR) assay, a Bartonella spp. nuoG gene fragment was detected in 1.51% (5/330) of the samples: 4 six‐banded armadillos (Euphractus sexcinctus) sampled in the MS and 1 southern tamandua (Tamandua tetradactyla) sampled in the PA. Eight sequences (5 ftsZ, 2 gltA and 1 rpoB) were obtained in the conventional PCR assays. In both phylogenetic analyses based on Bayesian and distance (SplitsTree) methods, the obtained ftsZ, gltA and rpoB sequences were positioned in a distinct clade, but related to B. washoensis. The analysis of SplitsTree and genotype networks based on B. washoensis sequences from several hosts from various localities of the world showed that the sequences of the present study were allocated in a group separated from the other sequences, indicating that they probably originated from median vectors and large numbers of mutational events. Additionally, the analyses performed by BLAST showed low percentages of identities of the sequences obtained in the present study when compared to those previously deposited in GenBank. Therefore, we propose a new Candidatus to Bartonella occurring in Xenarthra in Brazil. The present study was the first to report the occurrence of Bartonella sp. in mammals of the superorder Xenarthra in the world, and it was the first to describe a new Candidatus related to B. washoensis in Brazil.
Although mammals of the superorder Xenarthra are considered hosts of a wide range of zoonotic agents, works aiming at investigating the role of these animals as hosts for bacteria with zoonotic potential are rare. The present study aimed to investigate the occurrence and molecularly characterize Coxiella burnetii and haemoplasma (haemotropic mycoplasmas) DNA in blood and spleen samples from 397 free-living Xenarthra mammals (233 sloths, 107 anteaters and 57 armadillos) in five Brazilian states (Mato Grosso do Sul, São Paulo, Pará, Rondônia and Rio Grande do Sul). All biological samples from Xenarthra were negative in the qPCR for Coxiella burnetii based on the IS1111 gene. The absence of C. burnetii DNA in blood and spleen samples from Xenarthra suggests that these mammals may not act as possible hosts for this agent in the locations studied. When performed conventional PCR assays for the endogenous (gapdh) mammalian gene, 386 samples were positive. When screened by molecular assays based on the 16S rRNA gene of haemoplasmas, 81 samples were positive, of which 15.54% (60/386) were positive by conventional PCR and 5.44% (21/386) were positive by real-time PCR; three samples were positive in both assays.Of these, 39.74% (31/78) were also positive for the 23S rRNA gene and 7.69% (6/78) for the haemoplasma RNAse P gene. Among the samples positive for haemoplasmas, 25.64% (20/78) were obtained from anteaters (Tamandua tetradactyla and Myrmecophaga tridactyla), 39.74% (31/78) from sloths (Bradypus tridactylus, Bradypus sp. and Choloepus sp.) 34.61% (27/78) from armadillos (Priodontes maximus, Euphractus sexcinctus and Dasypus novemcinctus). A haemoplasma 16S rRNA sequence closely related and showing high identity (99.7%) to Mycoplasma wenyonii was detected, for the first time, in B. tridactylus. Based on the low identity and phylogenetic positioning of 16S rRNA and 23S rRNA sequences of haemoplasmas detected in anteaters and armadillos, the present study showed, for the first time, the occurrence of putative new
A magnaordem Xenarthra compreende um grupo de animais que divergiu há ~65 m.a. geograficamente isolado no hemisfério sul, atual América do Sul até o fechamento do Istmo do Panamá (~3 m.a), quando ocorreu um grande intercâmbio de animais entre as Américas do Norte e do Sul e uma variedade de Xenarthra migrou para o norte. Atualmente, Xenarthra das ordens Cingulata (tatus) e Pilosa (preguiças e tamanduás) são amplamente distribuídas na América do Sul e Central, enquanto na América do Norte, apenas tatus são encontrados. A história evolutiva desse grupo de animais, que é um dos mais antigos e endêmico da América do Sul, e a presença de diferentes espécies habitando uma diversidade de estratos e nichos ecológicos, fazem desse grupo um dos mais interessantes e menos explorados para estudos de diversidade, filogenia e aspectos biogeográficos de tripanossomatídeos. Esse estudo é de grande importância para o entendimento da diversidade e evolução dessa família. Estudos filogeográficos têm permitido inferir cenários que refletem as interações entre esses parasitas e seus hospedeiros vertebrados e vetores ao longo de suas histórias evolutivas. Nesse trabalho, foram analisadas 407 amostras de sangue e tecidos de Xenarthra da Amazônia brasileira (Rondônia, Pará e Mato Grosso), sendo 361 de preguiças, 32 de tamanduás e 14 de tatus. A fim de comparar o repertório de tripanossomatídeos encontrados nesses animais, com parasitas detectados em outras ordens de mamíferos, que compartilham o mesmo bioma, analisamos 57 amostras de porco-espinho (roedor do gênero Coendou). As amostras obtidas foram caracterizadas por DNA barcoding e análises filogenéticas foram realizadas baseadas nos genes SSU rRNA e gGAPDH, que permitem classificar os tripanossomatídeos em gêneros, subgêneros, espécies e genótipos conhecidos, assim como identificar novas espécies e genótipos. Os resultados obtidos revelaram uma grande diversidade de parasitas. Foram encontradas preguiças infectadas (34,35%) com os seguintes parasitas: Endotrypanum (N=54), duas novas espécies de Trypanosoma identificadas nesse estudo [T. sp1 (N=9) e T. sp2 (N=53)], T. rangeli das linhagens A (N=4), B (N=9) e E (N=3) e T. cruzi da DTU TcIII (N=2). As duas novas espécies de Trypanosoma foram detectadas em 86 animais: T. sp1, até o momento exclusivo de preguiças (9 animais), e T. sp2, detectado em todas as espécies de Xenarthra analisadas e também em espécies de Coendou. Nos tamanduás (65,63% infectados), foram detectados T. sp2 (N=18), T. rangeli das linhagens A (N=1), B (N=2) e E (N=1). Nos tatus (42,86% infectados), foram identificados T. sp2 (N=3) e T. cruzi III (N=4). Nossos resultados corroboraram estudos anteriores indicando uma forte associação do gênero Endotrypanum com preguiças, os únicos animais encontrados infectados por espécies desse gênero no nosso estudo. O encontro de T. cruzi TcIII em tatus também corrobora estudos prévios sugerindo uma forte associação desses animais com essa DTU. Além disso, pela primeira vez, TcIII foi identificado em preguiças, expandindo o número...
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