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.
This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine DiOC6(3). ...DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.
RESUMO: Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina DiOC6 (3) de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.
Xenarthra mammals can be found from southern North America to southern South America, including all Brazilian biomes. Although it has been shown that Xenarthra mammals can play a role as reservoirs ...for several zoonotic agents, few studies investigate the diversity of piroplasmids (Apicomplexa: Piroplasmida) in this group of mammals. Taking into account that piroplasmids can cause disease in animals and humans, understanding the prevalence and diversity of piroplasmids in Xenarthra mammals would contribute to conservation efforts for this group of animals as well as to infer risk areas for transmission of emergent zoonosis. The present study aimed to investigate the occurrence and molecular identity of piroplasmids in free-living mammals of the Superorder Xenarthra from four Brazilian states (Mato Grosso do Sul, São Paulo, Rondônia, and Pará). For this, DNA was extracted from blood or spleen samples from 455 animals. A nested PCR based on the 18S rRNA gene was used as screening for piroplasmids. Of the 455 samples analyzed, 25 (5.5%) were positive. Additionally, PCR assays based on 18S rRNA near-complete,
cox-1
,
cox-3
,
hsp70
,
cytB
,
β-tubulin
genes and the ITS-1 intergenic region were performed. Five out of 25 positive samples also tested positive for ITS-1-based PCR. The phylogenetic analysis positioned three 18S rRNA sequences detected in
Priodontes maximus
into the same clade of
Babesia
sp. detected in marsupials (
Didelphis albiventris
,
Didelphis marsupialis
, and
Monodelphis domestica
) and
Amblyomma dubitatum
collected from opossums and coatis in Brazil. On the other hand, the 18S rRNA sequence obtained from
Dasypus novemcinctus
was closely related to a
Theileria
sp. sequence previously detected in armadillos from Mato Grosso State, grouping in a subclade within the
Theileria sensu stricto
clade. In the phylogenetic analysis based on the ITS-1 region, the sequences obtained from
Myrmecophaga tridactyla
and
Tamandua tetradactyla
were placed into a single clade, apart from the other piroplasmid clades. The present study demonstrated the molecular occurrence of Piroplasmida in anteaters and
Babesia
sp. and
Theileria
sp. in armadillos from Brazil.
Digenetic eucotylids of the Paratanaisia genus are widely reported parasites of the avian renal system. The infection, commonly reported in free-living and zoo-housed birds, is described for the ...first time in a domiciled bird, a cockatiel kept as pet bird with free access to the yard outdoors. The bird was received at Veterinary Hospital, where clinical and radiographic evaluations suggested a case of heavy-metal poisoning. Although the bird received supportive care and a chelating agent, it died the next day. The necropsy showed friable kidneys and congestion of blood vessels in structures such as the skin, proventriculus, brain, and skullcap. The histopathological evaluation of the kidneys revealed multifocal hemorrhages, commonly found in cases of heavy-metal poisoning. Parasitic structures similar to those of the digenean trematodes Paratanaisia spp. were also observed inside dilated collecting ducts, which presented epithelial cell flattening and vacuolization. There was compression of adjacent tissue and discrete fibrotic areas. In the presence of intermediate hosts in the yard, the synanthropic nature of some wild free-living birds could make them a source of trematodes infection and dissemination for pet birds. Conversely, the infected cockatiel could also have served as a reservoir and carrier of trematodes to wild free-living birds.
Canine monocytic ehrlichiosis (CME) is one of the most important tick-borne diseases worldwide, with multisystemic presentations. Immune dysregulation has been proposed as the primary mechanism ...involved in its pathogenesis and in tissue injury in dogs with CME. Experimental infection of German Shepherd dogs in the present study demonstrated that CME caused marked pathological changes in their lymph nodes and spleen, and also gave rise to mononuclear infiltration in organs and tissues. Immunophenotyping of cells in lymph nodes, spleen and injured tissues highlighted differences in lymphocyte subsets, local expression of immunoglobulin subclasses and MHCII molecules between infected and control dogs. These findings suggest that the immunophenotypic and immunopathological changes in dogs with acute experimental CME are related to Th1 bias and compartmentalized immune response.
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 Candidatus haemotropic Mycoplasma spp. (“Candidatus Mycoplasma haematotetradactyla” and “Candidatus Mycoplasma haematomaximus”) in Xenarthra mammals from Brazil.
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.
Toxoplasma gondii and Neospora caninum are Apicomplexan intracellular protozoan parasites that affect numerous animal species, thus leading to severe diseases and economic losses, depending on the ...vertebrate species involved. The role of the avian species in maintaining and transmission of these coccidia has been studied for several years as they tend to serve as a potential source of infection for mammals and humans. The present study aimed to assess the serological exposure of Orinoco goose (Neochen jubata) to T. gondii and N. caninum. Between 2010 and 2013, 41 free-ranging Orinoco geese were captured in the Araguaia River, Brazil. The presence and titration of IgY antibodies to both coccidia were assayed via indirect immunofluorescent antibody test (IFAT). While IgY antibodies for N. caninum were present in 5 animals, with titers of 20, the antibodies for T. gondii were found in 35 animals, with titers ranging from 20 to 640. Considering that the Orinoco goose's meat is consumed by the local population in the studied area, it may represent an important source of T. gondii infection for humans. Due to its migratory behavior, this goose may play a pivotal role in the natural dispersion of both parasites. Furthermore, molecular studies are required for genotyping the isolates of T. gondii that occurs in this avian species.
This report presents a description of natural infection of an adult free-living guira cuckoo with Paratanaisia confusa. Histological and morphological evaluations of renal and parasite samples were ...performed. The morphological analysis of the parasites revealed spiny tegument, characteristic of P. confusa helminthes. Although macroscopic alterations were absent in the kidneys, the histopathology revealed parasites inside the collecting ducts causing dilatation and destruction of the lining epithelial cells in addition to small focal inflammatory infiltrates in the renal parenchyma. However, as the bird was free-living and naturally infected it was not possible to confirm if all these alterations were directly caused by the parasite presence or if they were related to causes other than the parasitism. Nonetheless, such findings indicate that these trematodes may have pathogenic potential in this host.
Over the past years, the number of neoplasm cases reported in nonhuman primates have considerably increased, with hepatocellular tumors reported in several species. Here a case report of extrahepatic ...cholangiocarcinoma in an adult male Black Howler Monkey (Alouatta caraya) is presented. The animal, kept as pet, was admitted at Veterinary Hospital presented clinical signs of cholestasis. After unsuccessful drug treatment, the animal was submitted to a cholecystoduodenostomy. During the surgical procedure, the hepatobiliary system was meticulously observed. The liver showed no macroscopic alterations, however, there was a solid mass throughout the common bile duct. Biopsy specimens were collected from the bile duct mass, liver and gallbladder for histological and immunohistochemical evaluations. Histological examination of the liver showed diffuse accentuated vacuolization and necrosis, besides hemosiderin accumulation inside hepatocytes. In the portal region, there was an accentuated proliferation of biliary ducts, which presented replete with bilirubin. Discrete neutrophilic inflammatory infiltrate and fibrosis were also observed. The bile duct mass showed a cellular proliferation of epithelial origin, confirmed by immunohistochemical analysis. The arrangement, characteristics and growth pattern of these cells revealed a malignant neoplasm. Despite the success of the procedure, the animal died hours later. Cholangiocarcinomas generally present high mortality rates, since its late clinical presentation make the treatments ineffective due to its large extensions at the time of diagnosis. The owner did not allow the necropsy, therefore the presence of other neoplasms and important organ alterations, which may be aggravated the patient's clinical condition, were not confirmed.
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