Post-mortem detection of hemoplasmas (hemotropic Mycoplasma spp.) in South American fur seal (Arctocephalus australis) sampled in Rio Grande do Sul State, southern Brazil.

Hemotropic mycoplasmas are bacteria that attaches to erythrocytes surface, which some species presents zoonotic concerns. In the suborder Pinnipedia, genera Otaria and Arctocephalus are prominent in Brazil. This study investigated the occurrence of hemoplasmas in Arctocephalus sp. and Otaria flavescens found dead along the coast of a Southern Brazilian State. DNA from 135 spleen samples were extracted and subjected to conventional PCR protocols, targeting the 16 S rRNA and 23 S rRNA gene. Three (2.22 %) Arctocephalus australis were positive in the 16 S rRNA gene, and no samples amplified in the 23 S rRNA gene. Samples from this study clustered with Zalophus californianus and Arctocephalus tropicalis mycoplasmas on a Bayesian phylogenetic analysis. Genetic diversity analysis suggested distinct genotypes, indicating A. australis as a new host for hemoplasma, and also a potential putative novel hemoplasma genotype. These findings raises future awareness for pinnipeds conservation, and adds Mycoplasma spp. to be taken into consideration when clinically evaluating rescued animals.

Pathological, morphological, and molecular characterization of the infection by Brachycladium goliath in humpback whale Megaptera novaeangliae.

The humpback whale (Megaptera novaeangliae) is a cosmopolitan migratory, seasonal mysticete that frequents the Brazilian coast. Strands of specimens may occur during the migratory stay in the country. In 2021 and 2022, three live humpback whales stranded on the coast of Rio Grande do Sul and Santa Catarina states in southern Brazil. After euthanasia, specimens were necropsied, and organs were thoroughly examined for lesions. Grossly, in all three cases, the liver exhibited multifocal, irregular, firm, white areas on the hepatic capsule, which extended into the parenchyma. On the cut surface, the livers were yellow to pale brown with orangish to greenish areas, the bile ducts were prominent, thickened, and severely dilated, and leaf-shaped flukes were found inside of them. Additionally, one case showed moderate atrophy of the right hepatic lobe. The histological findings included dilation of bile ducts, hyperplasia of the bile duct epithelium, marked inflammatory infiltration of lymphocytes, plasma cells, and eosinophils, and portal fibrosis. The parasite Brachycladium goliath was both morphologically and molecularly identified based on diagnostic key for trematodes and the original description of the species, and the amplification and sequencing of the ITS-2 region, respectively. Even though hepatic injury was not the primary cause of stranding, it may have contributed to the debilitation of the whales. To the authors' knowledge, this is the first study that reports M. novaeangliae as a definitive host of B. goliath and that describes the lesions caused by the parasite in cetaceans.

Dirofilaria immitis and Onchocercidae spp. in wild felids from Brazil.

Among the species described within the Onchocercidae family, Dirofilaria immitis is regarded as the most common worldwide, causing severe and often fatal conditions in dogs, cats, and occasionally humans. Dirofilaria spp. are vectored by mosquitoes, simulids, and culicoids, with their epidemiology dependent on the geographical distribution of competent vectors. Eight species of Dirofilaria have been reported so far in Brazil, of which six parasitize non-human primates, deer, procyonids, and marsupials. Here, we investigated the occurrence of Onchocercidae in wild felids (i.e., Panthera onca, Puma concolor, Herpailurus yagouaroundi, Leopardus geoffroyi, Leopardus guttulus, Leopardus pardalis, Leopardus wiedii, Leopardus munoai) from different locations in Brazil. Overall, 82 samples (n = 63 blood; n = 19 tissues) were molecularly screened for cytochrome c oxidase subunit-1 (cox1) gene. Four (i.e., 4.8%) wild felid samples were positive, and at BLAST analysis, the obtained sequences showed varying percentage of nucleotide identity with the genera Brugia (i.e., 87-88%), Setaria (i.e., 89%), and D. immitis (i.e., 94.4%). Phylogenetic analyses clustered sequences obtained into three distinct clades, one with D. immitis and the remaining two with other Onchocercidae spp. Data herein obtained highlight the need for a more comprehensive understanding of the diversity and biology of Onchocercidae in South America in order to assess the potential impact that these species may have for domestic and wild animals, as well as humans.

Eyeworms of wild birds and new record of Thelazia (Thelaziella) aquilina (Nematoda: Spirurida).

The eyeworms of the subgenus Thelaziella infect orbital cavities, conjunctival sacs and lachrymal ducts of many wild birds, being able to cause conjunctivitis. In Brazil, at least 10 species of Thelaziella have been described. Here we present a brief review of cases of Thelazia (Thelaziella) aquilina and describe the infection of this parasite in Harpia harpyja, as a new host record. Two nematodes were collected from the surface of the cornea of an adult female H. harpyja in the Amazon region (Brazil) and were morphologically identified as two males of T. (T.) aquilina. This data demonstrated the need for further studies focused on elucidating the transmission, biological cycle, and phylogenetic positioning of Thelaziella as subgenus.

Psoralges libertus mite infestation in Tamandua tetradactyla: a 127-year gap in the literature due to scarcity of reports.

This study addresses the report of Psoralges libertus mite infestations in Southern Tamandua (Tamandua tetradactyla) after a 127-year gap in the literature, detailing clinical conditions, histopathological findings, and mite identification via morphology. The research broadens our understanding of mite habitats, highlighting challenges in managing parasitic health in free-ranging and captive animals. It underscores the importance of ongoing surveillance for conservation and public health, including potential zoonotic concerns. A P. libertus report in T. tetradactyla after over a century highlights the scarcity of publications on mite-induced mange in this mammalian species. Investigating potential P. libertus' impacts on tamanduas' well-being necessitates further research, particularly regarding pathogenicity, agent epidemiology, and host interaction. This study enhances our comprehension of parasite-host interactions and their relevance to wildlife conservation.

A long-term study on free-ranging jaguar-tick interactions, featuring a novel report of Amblyomma incisum adult infestation.

Over nearly 12 years, we collected ticks from free-ranging jaguars (Panthera onca) and performed statistical analyses to comprehend the vector-host relationship throughout the seasons. We evaluated the presence and number of ticks, as well as their association with weight, age, and gender of captured jaguars in the Pantanal and Amazon biomes. Out of 100 captured jaguars (comprising 72 initial captures and 28 recaptures, with 41 females and 31 males), 77 were found to be infested by different tick species. We gathered a total of 1,002 ticks, categorized by the following species in descending order of abundance: Amblyomma sculptum, Amblyomma ovale, Rhipicephalus microplus, Amblyomma triste, Amblyomma cajennense sensu stricto, Amblyomma incisum, and Amblyomma spp. larvae. Apart from weight, statistical analysis indicated that age, gender and seasonality does not significantly affect the presence of different tick species in free ranging jaguars. Notably, A. sculptum adults were more abundant in the first semester, while A. sculptum nymphs and Amblyomma spp. larvae were mainly found during dry months, aligning with their expected life cycle stages. This is the first long-term study in jaguars to correlate seasonality and host factors and also the first time an adult of A. incisum is reported infesting a jaguar.

Detection of Trypanosoma evansi in jaguars (Panthera onca): insights from the Brazilian Pantanal wetland.

Trypanosoma evansi is a widespread and neglected zoonotic parasite that affects domestic and wild animals, causing a disease commonly known as "surra." The Brazilian Pantanal wetland is recognized as an enzootic area for this protozoan, yet recognizing the importance of reservoir hosts also in order to prevent zoonotic outbreaks. This study aimed to assess the occurrence of T. evansi in jaguars (Panthera onca) from the Brazilian Pantanal wetland and explore associated clinical and hematological manifestations. A total of 42 animals were screened by PCR and sequenced for species identification when positive. Trypanosoma evansi was detected in six free-ranging jaguars (six positive animals of 42 captures and 16 recaptures), representing the first molecular evidence of such infection in this animal species. Our findings suggest that jaguars may act as reservoir hosts of T. evansi in the Brazilian Pantanal wetland. The better understanding of the role of wildlife in the epidemiology of T. evansi is also of importance to future reintroduction and translocation programs toward wildlife conservation efforts.

The jaguar (Panthera onca) as a potential new host of Dracunculus sp.

Nematode species of the genus Dracunculus (Spirurida: Dracunculoidea) infect tissues and body cavities of reptiles, domestic and wild carnivores, and humans. The definitive hosts acquire the infection by ingesting intermediate (i.e., cyclopoid copepod) or paratenic (i.e., amphibians and fishes) hosts. Here we report the jaguar (Panthera onca) as a potential new host for Dracunculus sp. The nematode was collected from an ulcerated cutaneous nodule on the left anterior limb of a female jaguar in the municipality of Miranda, Mato Grosso do Sul state, Brazil. Based on the morphology of first stage larvae collected from a small fragment of the uterus of the adult nematode, the species was identified as Dracunculus sp. Reichard, 1759. Additionally, the morphological identification was molecularly confirmed by sequencing the cox1 gene. This report advocates for further investigations into the transmission cycle of this parasite in the Brazilian Pantanal wetland, considering the role of wildlife hosts and the zoonotic potential of Dracunculus species in that area.

Pathological findings associated with Dipetalonema spp. (Spirurida, Onchocercidae) infection in two species of Neotropical monkeys from Brazil.

Among vector-borne helminths, filarioids of the genus Dipetalonema (Spirurida: Onchocercidae) localize in several tissues and body cavities of several animal species, causing mild to moderate lesions. The pathological findings associated with Dipetalonema spp. infection in Neotropical monkeys from southern Brazil are herein described, along with a fatal case due to filarial polyserositis and entrapment of an intestinal segment. At necropsy, nematodes were observed in abdominal and thoracic cavities, or in the pericardium of 37 (31.3%) out of the 118 individuals examined (i.e., 35 Alouatta guariba clamitans and two Sapajus nigritus). In addition, at histology, 27.0% of positive animals presented microfilarie (inside blood vessels of lung, spleen, liver, and brain) and 8.1% presented adult nematodes in the heart, lung, and liver. In two cases, cross-sections of filarioids were associated with areas of epicardial thickening with intense fibrosis and pyogranulomatous inflammation in the brain, heart, liver, lungs, or spleen. The DNA fragment was amplify using the cox1 gene, sequenced and analyzed to identify the nematode species collected; presence of Wolbachia was assessed in the filarioids using the 16S rRNA gene. At BLAST analysis of the cox1 gene, 10 sequences showed 91.7% nucleotide identity with Dipetalonema gracile, and two with D. gracile (98.5%) and Dipetalonema graciliformis (98.3%). Phylogenetic analyses clustered sequences of the cox1 obtained in this study in two clades corresponding with the host species. Wolbachia sp. endosymbiont was detected in four samples. Data herein reported provide a description of pathological lesions associated with the infection by Dipetalonema spp., suggesting that they may cause disease in Neotropical monkeys. In addition, a better understanding of diversity and biology of Dipetalonema spp. in South America is needed to assess the impact they may cause in native non-human primates from Brazil.

Viability of erythrocytes in canine packed red blood cells stored in CPDA-1 is related to the presence of Mycoplasma haemocanis.

Hemotropic mycoplasmas are associated with subclinical disease in dogs and should be identified in blood donors. The objective was to investigate the presence and effect of M. haemocanis in units of packed red blood cells (pRBC) during storage. Canine donors (n = 10) were screened for M. haemocanis by quantitative real-time PCR. pRBCs were obtained from 5 hemoplasma negative dogs and 5 hemoplasma positive dogs. Each pRBC was aliquoted into two 100 mL transfer bags and stored at 4 °C. M. haemocanis loads and biochemical variables (pH, bicarbonate, potassium, sodium, chlorite, glucose, lactate, ammonia, PCV, and % hemolysis) were evaluated on days 1, 7, 18, and 29. M. haemocanis loads increased in pRBC from day 1-29 of storage. Glucose decreased and lactate increase faster in pRBC with M. haemocanis. This study contributes to understand hemoplasma metabolism and reinforces that dog donors should be tested for hemoplasmas.

Molecular survey of hemotropic mycoplasmas in crab-eating raccoons (Procyon cancrivorus) in southern Brazil.

Hemoplasmas are non-cultivable bacterial parasites of erythrocytes that infect domestic and wild animals, as well as humans. Their means of transmission and pathogenesis remain contentious issues and difficult to evaluate in wild animals. Procyon cancrivorus is a South American carnivore and occurs in all Brazilian biomes. In this study, we aimed to investigate occurrences of hemoplasmas infecting P. cancrivorus and to identify their 16S rRNA gene, in southern Brazil. DNA was extracted from spleen and blood samples of P. cancrivorus (n = 9) from different locations. Hemoplasma DNA was detected in six samples, based on 16S rRNA gene amplification and phylogenetic analysis. Four of the six sequences belonged to the "Mycoplasma haemofelis group", which is closely related to genotypes detected in Procyon lotor from the USA; one was within the "Mycoplasma suis group", closely related to "Candidatus Mycoplasma haemominutum"; and one was within the intermediate group between these clusters. Thus, these sequences showed that the molecular identity of hemoplasmas in the population studied was very variable. In five positive animals, Amblyomma aureolatum ticks and a flea (Ctenocephalides felis felis) were collected. The present study describes the first molecular detection of mycoplasmas in P. cancrivorus.

Hematologic variables of free-living Leptodactylus luctator with and without hemoparasites and thrombidiform mites in southern Brazil.

It has been suggested that anuran amphibian parasites can cause clinical signs in situations of environmental imbalance. In the family Leptodactylidae, information about hematology is scarce, although these are well-known tools for the diagnosis and prognosis in clinical practice and potential bioindicators of environmental stress. The objective of this study is to describe Leptodactylus luctator hematology, to report the occurrence of hemoparasites and thrombidiform mites, and to compare the hematological variables under the presence and absence of these organisms. Ectoparasites and heparinized blood samples from 40 free-living specimens of L. luctator were collected for analysis. Hematologic variables and total plasma protein were compared between groups with and without hemoparasites and intradermal mites. As results, structures compatible with hemogregarines, Lankesterella sp., five morphotypes of Trypanosoma spp., microfilaria, Aegyptianella sp., an unidentified intraleukocytic hemoparasite, and frog erythrocytic virus (FEV) inclusion bodies were identified in the blood samples, besides Hannemania spp. intradermal mites. The hemoparasite occurrence was higher than previously reported in other anuran families and locations. Also, L. luctator has smaller red blood cells (RBCs) and white blood cells (WBCs), and a hyposegmentation of the neutrophil nucleus, when compared to many other amphibians. White blood cell, neutrophil, and monocyte counts were higher in animals parasitized by mites. There was no correlation between the number of parasitized RBCs and hematologic variables. This study provides anuran hematologic information, in addition to indicating a host reaction to infestation by Hannemania spp. mites, besides constituting the first record of the distribution of hemoparasites and intradermal mites in L. luctator of the study region.

Emergence of Mansonella sp. in free-ranging primates in southern Brazil.

Mansonellosis is a neglected and emerging tropical disease. Among all zoonotic filarial diseases, it is probably the most prevalent and least studied, with approximately 114 million people infected. The parasites of Mansonella spp. are among the most common blood parasitemias and are widely found in Africa and Latin America. Through molecular analysis of blood samples from free-ranging primates Sapajus nigritus (n 33) and Alouatta guariba clamitans (n 5) in the southern states of Brazil (Santa Catarina and Rio Grande do Sul), we identified samples positive for Mansonella perstans in two specimens of A. guariba clamitans. A fragment of 578 bp from the ITS intergenic region (5.8S-ITS2-28S) was targeted for an initial PCR screening. Subsequently, positive samples were subjected to other PCR assays targeting a fragment of the 12S and the 18S genes. This is the first record of molecular detection of the agent in this host in the Pampa Biome. With a wide distribution across Brazil and Argentina, these primates may represent a potential wild reservoir for the zoonotic agent of mansonellosis. Entomological and transmission studies are essential to avoid the urbanization of mansonellosis and to understand the cycles of agents in different environmental scenarios.

First record of melanism in the critically endangered Pampa cat (Leopardus munoai), an endemic species of the Pampa grasslands / Primeiro registro de melanismo no criticamente ameaçado gato-pampeano (Leopardus munoai), uma espécie endêmica das áreas abertas do Pampa

Abstract We report the first record of a melanistic individual of the critically endangered Pampa cat (Leopardus munoai), from July 8th, 2021, at 10:45 am (coordinates 30.096288° S; 54.941139° W) in the area of the Brazilian army, known as Campo de Instrução Barão de São Borja (CIBSB), popularly known as Saicã.

Resumo: Este trabalho reporta o primeiro registro de melanismo do criticamente ameaçado de extinção gato-pampeano (Leopardus munoai), realizado em 8 de julho de 2021, às 10:45 da manhã. O registro foi obtido nas coordenadas 30.096288° S; 54.941139° W na área do exército brasileiro Campo de Instrução Barão de São Borja (CIBSB), popularmente conhecida como Saicã.

Theileria terrestris nov. sp.: A Novel Theileria in Lowland Tapirs (Tapirus terrestris) from Two Different Biomes in Brazil.

The low-land tapir (Tapirus terrestris) is the largest wild terrestrial mammal found in Brazil. Although T. terrestris has been already reported as a host of hemoparasites, the occurrence and genetic identity of Piroplasmida agents in this species is still cloudy. Although it is reported that Theileria equi, an endemic equid-infective agent in Brazil, is occurring in lowland tapirs, these reports are probably misconceived diagnoses since they are solely based on small fragments of 18S rRNA that may not achieve accurate topologies on phylogenetic analyses. The present study aimed to detect and investigate the identity of Theileria spp. in tapirs from Pantanal and Cerrado biomes. Blood-DNA samples from tapirs were screened for a partial (~800 bp) 18S rRNA gene fragment from Piroplasmida and 64 (64/122; 52.46% CI: 43.66-61.11%) presented bands of expected size. Samples were submitted to different protocols for molecular characterization, including near-full length 18S rRNA gene (~1500 bp), and the ema-1 gene from T. equi. Eight sequences were obtained for extended fragments (1182-1473 bp) from the 18S rRNA gene. Moreover, three sequences from partial cox-1 and five from partial hsp70 gene were obtained. None of the samples presented amplifications for the ema-1 gene. Phylogenetic and distance analyses from the 18S rRNA sequences obtained demonstrated a clear separation from tapirs' Theileria spp. and T. equi. Phylogenetic analyses of cox-1 and hsp70 sequences obtained herein also showed a unique clade formed by tapir's Theileria spp. Theileria terrestris sp. nov. is positioned apart from all other Theileria species in 18S rRNA, cox-1, and hps70 phylogenetic analyses. This novel proposed species represents a new Piroplasmida clade, yet to be characterized regarding biological features, vectors involved in the transmission cycles, additional vertebrate hosts, and pathogenicity.

First report of canine rangeliosis in domestic dogs from different regions of Santa Catarina State, Brazil.

Rangeliosis is the disease caused by Rangelia vitalii, a parasite reported in dogs from southeastern and southern Brazil, Uruguay, Paraguay, and Argentina. This protozoan is transmitted by the ixodid Amblyomma aureolatum, and infects erythrocytes, leukocytes, and vascular endothelial cells of the host. The common clinical signs, such as prostration, fever, anemia, thrombocytopenia, anorexia, weight loss, and dehydration, are also found in other infections, like canine babesiosis and ehrlichiosis. The similar clinical presentation with other diseases, as well as the indistinguishable morphology with intraerythrocytic Babesia canis, complicates the disease diagnostic. In the present study, blood samples from dogs presenting clinical signs compatible with hemoparasitosis were investigated for rangeliosis. The dogs were treated at veterinary clinics in the cities of Blumenau and Lages, in the State of Santa Catarina, Brazil. Blood samples from 17 dogs were analyzed by PCR. The samples were screened by a conventional piroplasma-PCR and the positives confirmed by a specific R. vitalii-qPCR. Two animals (2/17; 11.8%) were positive for R. vitalii, one from Blumenau and the other from Lages. Both animals presented unspecific signs of hemoparasitosis, such as apathy, anemia, and anorexia. The results indicate the necessity of molecular assays for the proper identification of the hemoparasite, and to investigate the real prevalence of rangeliosis in the State of Santa Catarina.

Prevalence and molecular detection of Leishmania spp. in bats from Rio Grande do Sul state, Brazil.

This study aimed to detect the occurrence of infection by Leishmania spp.in bats from 34 municipalities of Rio Grande do Sul state (RS; southern Brazil) from 2016 to 2021. A total of 109 bats were provided by the Centro Estadual de Vigilância em Saúde of RS, including six species belonged to Molossidae family, six to Vespertilionidae family, and two to Phyllostomidae family. Leishmania spp. was identified using the nested-PCR method by amplifying the SSU rDNA ribosomal subunit gene into four organ pools: (1) the liver, spleen, and lymph node; (2) heart and lungs; (3) skin; and (4) bone marrow of each bat. Three (3/109, 2.7%) animals tested positive for Leishmania spp. The respective PCR-positive organs came from pools 1 and 3. Two bats (Tadarida brasiliensis) were from the municipality of Canoas, and sequences analysis confirms the species identification as Leishmania infantum. In the third bat (Molossus molossus), from Rio Grande, it was not possible to determine the protozoa species, being considered Leishmania spp. Our results indicate that bats can participate in the biological cycle of Leishmania spp. and perform as host, reservoir, and/or source of infection of the protozoa in different areas of RS. More studies will be needed to elucidate the role of these Chiropteras in the circulation of Leishmania spp. This is the first study reporting the occurrence of Leishmania spp. in bats in Rio Grande do Sul state, southern Brazil.

Epidemiological compatibility of Amblyomma sculptum as possible vector and Panthera onca as reservoir of Cytauxzoon spp. in Midwestern Brazil.

Cytauxzoonosis is an acute and highly lethal tick-borne disease of wild and domestic cats, and is widely distributed in Africa, Asia, Europe, the USA and Brazil. So far, only two tick species present on the USA are experimentally confirmed in Cytauxzoon transmission however, in Brazil and other continents, the epidemiology of the disease remains unknown. Evidences points to Panthera onca as a possible reservoir, but there is no evidence to point the vector. Therefore, this study evaluates the presence of Cytauxzoon spp. in wild felids from areas with and without records of Amblyomma sculptum this ixodid for comparison. Overall, 53 blood samples of P. onca, Puma concolor, and Leopardus pardalis from the Midwest region (MR; region with A. sculptum) and 143 blood and/or spleen samples from Leopardus geoffroyi, Leopardus wiedii, Leopardus munoai, Leopardus guttulus, Herpailurus yagouaroundi, L. pardalis, and P. concolor from Rio Grande do Sul State (RS; without A. sculptum). Only one feline sample was negative for Cytauxzoon sp. from MR; no samples from RS were positive. In total, 507 ticks were identified from MR felids, with predominance of A. sculptum (69.23%). In RS, there were 93 ixodids, of which 90.32% were Amblyomma aureolatum. The difference in the tick fauna of the two regions studied (presence/absence of A. sculptum) reflects the results found. This study highlighted A. sculptum as a possible vector since this hemoparasite was abundantly observed in areas where it occurs, also, there was no evidence of Cytauxzoon spp. where it was absent. Additionally, the study supported the suggestion that P. onca is the reservoir for the agent in MR.

Rickettsia amblyommatis-infected Amblyomma coelebs parasitizing a human traveler in Rio Grande do Sul, southern Brazil, after returning from the Amazon.

With increased national and international transportation, humans as well as domestic and wild animals are able to get from one end to the other within hours, accidently carrying transboundary vectors and pathogens. We report here, the introduction of Amblyomma coelebs in Rio Grande do Sul, the southernmost state of Brazil, via a man travelling to the Carajás National Forest in the state of Pará (2661 km away). Upon return to his residence in Porto Alegre, RS, he realized that he was parasitized by one tick. The tick was identified as a nymph of Amblyomma sp., placed in a Biological Oxygen Demand incubator, and after ecdysis was identified as a male of Amblyomma coelebs. The tick's DNA was screened for Rickettsia spp., where Rickettsia amblyommatis DNA was detected. The parasitized man was seropositive to spotted fever group rickettsiae, by immunofluorescence assay, in a single serum sample collected 45 days after the tick-bite. Amblyomma coelebs is found in some Brazilian states, but Parana is the southernmost state in Brazil where it occurs. This species has already been reported to carry some species of Rickettsia, including species of the spotted fever group. It may risk public health if established in Rio Grande do Sul, with the introduction of tick and pathogenic rickettsias.

Molecular investigation of haemotropic mycoplasmas and Coxiella burnetii in free-living Xenarthra mammals from Brazil, with evidence of new haemoplasma species.

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.