First molecular detection of Borrelia sp. in tapirs (Tapirus terrestris).

Borrelia theileri is a tick-borne spirochete causative agent of fever, apathy and reduced food consumption in cattle. Molecular diagnosis has expanded the understanding of Borrelia theileri with new hosts and geographical locations being described. The present study aimed to describe the first molecular detection of B. theileri in wild tapirs (Tapirus terrestris) from South America. Blood DNA samples obtained from 99 tapirs sampled in Pantanal (n = 61) and Cerrado (n = 38) biomes were screened using a qPCR assay based on the 16 S rRNA gene of Borrelia sp. Positive samples in the qPCR assay were subjected to PCR assays to allow characterization of fragments from 16 S rRNA and flaB genes. Two (2/99; 2.0%) animals from Pantanal biome were positive in the qPCR and one sample presented bands of expected size for the flaB protocol. Amplicons from this sample were successfully cloned and sequenced. In the phylogenetic analysis, Borrelia sp. from T. terrestris grouped together with B. theileri sequences previously detected in Rhipicephalus microplus ticks and cattle from Minas Gerais State in Brazil, Rhipicephalus geigyi from Mali, and R. microplus and Haemaphysalis sulcata from Pakistan. This finding contributes to our knowledge regarding susceptible hosts species for B. theileri. More studies are necessary to understand the potential effects of B. theileri on tapir's health.

Molecular evidence of Bartonella spp. in wild lowland tapirs (Tapirus terrestris), the largest land mammals in Brazil.

The genus Bartonella (Hyphomicrobiales: Bartonellaceae) encompasses facultative intracellular α-proteobacteria that parasite erythrocytes and endothelial cells from a wide range of vertebrate hosts and can cause disease in animals and humans. Considering the large diversity of vertebrate species that may act as reservoirs and arthropod species that may be associated with Bartonella transmission, the exposure of animals and humans to these microorganisms is likely underestimated. The present study aimed to investigate the occurrence of Bartonella sp. in wild tapirs (Tapirus terrestris; Perissodactyla: Tapiridae) from two biomes in Brazil: Pantanal and Cerrado. Ninety-nine GPS-monitored wild tapirs were sampled in Pantanal (n = 61/99) and Cerrado (n = 38/99). A qPCR (quantitative real-time polymerase chain reaction) assay targeting the nuoG gene was used for the screening for Bartonella spp. DNA. Positive samples were additionally subjected to conventional PCR assays targeting five molecular markers (ribC, gltA, rpoB, groEL, ITS). Eight (8/99; 08,08%) animals were positive in the qPCR assay for Bartonella spp.: 7 from Cerrado (7/8; 87.5%) and 1 from Pantanal (1/8; 12.5%). The 5 Bartonella ribC sequences obtained from tapirs' blood samples grouped together with Bartonella henselae obtained from cats, humans, wild felids and Ctenocephalides felis (Siphonaptera: Pulicidae) fleas. To the best of author's knowledge, this is the first report of Bartonella sp. in Tapirus terrestris. This finding contributes to the understanding of the occurrence of B henselae in wild mammals from Brazil as well as expands the knowledge regarding the potential vector-borne pathogens that may affect wild tapis from Cerrado and Pantanal biomes.

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.

Evaluating expert-based habitat suitability information of terrestrial mammals with GPS-tracking data.

Aim: Macroecological studies that require habitat suitability data for many species often derive this information from expert opinion. However, expert-based information is inherently subjective and thus prone to errors. The increasing availability of GPS tracking data offers opportunities to evaluate and supplement expert-based information with detailed empirical evidence. Here, we compared expert-based habitat suitability information from the International Union for Conservation of Nature (IUCN) with habitat suitability information derived from GPS-tracking data of 1,498 individuals from 49 mammal species. Location: Worldwide. Time period: 1998-2021. Major taxa studied: Forty-nine terrestrial mammal species. Methods: Using GPS data, we estimated two measures of habitat suitability for each individual animal: proportional habitat use (proportion of GPS locations within a habitat type), and selection ratio (habitat use relative to its availability). For each individual we then evaluated whether the GPS-based habitat suitability measures were in agreement with the IUCN data. To that end, we calculated the probability that the ranking of empirical habitat suitability measures was in agreement with IUCN's classification into suitable, marginal and unsuitable habitat types. Results: IUCN habitat suitability data were in accordance with the GPS data (> 95% probability of agreement) for 33 out of 49 species based on proportional habitat use estimates and for 25 out of 49 species based on selection ratios. In addition, 37 and 34 species had a > 50% probability of agreement based on proportional habitat use and selection ratios, respectively. Main conclusions: We show how GPS-tracking data can be used to evaluate IUCN habitat suitability data. Our findings indicate that for the majority of species included in this study, it is appropriate to use IUCN habitat suitability data in macroecological studies. Furthermore, we show that GPS-tracking data can be used to identify and prioritize species and habitat types for re-evaluation of IUCN habitat suitability data.

Expanding the Universe of Hemoplasmas: Multi-Locus Sequencing Reveals Putative Novel Hemoplasmas in Lowland Tapirs (Tapirus terrestris), the Largest Land Mammals in Brazil.

The lowland tapir (Tapirus terrestris) is the largest land mammal in Brazil and classified as a vulnerable species, according to the assessment of the risk of extinction. The present study aimed at investigating the occurrence and genetic diversity of hemoplasmas in free-ranging T. terrestris from the Brazilian Pantanal and Cerrado biomes. Blood samples were collected from 94 living and eight road-killed tapirs, totalizing 125 samples Conventional PCR targeting four different genes (16S rRNA, 23S rRNA, RNAse P, and dnaK) were performed, and the obtained sequences were submitted for phylogenetic, genotype diversity, and distance analyses. The association between hemoplasma positivity and possible risk variables (age, gender, and origin) was assessed. Out of 122 analyzed samples, 41 (41/122; 33.61% CI: 25.84-42.38%) were positive in the 16S rRNA-based PCR assay for hemoplasmas. Positivity for hemoplasmas did not differ between tapirs' gender and age. Tapirs from Pantanal were 5.64 times more likely to present positive results for hemoplasmas when compared to tapirs sampled in Cerrado. BLASTn, phylogenetic, genotype diversity, and distance analyses performed herein showed that the sampled lowland tapirs might be infected by two genetically distinct hemoplasmas, namely 'Candidatus Mycoplasma haematoterrestris' and 'Candidatus Mycoplasma haematotapirus'. While the former was positioned into "Mycoplasma haemofelis group" and closely related to 'Candidatus Mycoplasma haematoparvum, the latter was positioned into "Mycoplasma suis group" and closely related to 'Candidatus Mycoplasma haematobos'. The impact of both putative novel species on tapir health status should be investigated.

Rabies Virus Exposure in Wild Lowland Tapirs (Tapirus terrestris) from Three Brazilian Biomes.

We evaluated the presence of antibodies for rabies virus in 177 serum samples from 125 wild lowland tapirs (Tapirus terrestris) from three different Brazilian biomes. The rapid fluorescent focus inhibition test was performed. No antibody titers suggesting the circulation of the rabies virus in tapir habitat were detected.

Ticks and rickettsial exposure in lowland tapirs (Tapirus terrestris) of three Brazilian biomes.

This study evaluated ticks and rickettsial exposure in 220 free-ranging lowland tapirs, Tapirus terrestris, from 2006 to 2018 in selected areas of three major biomes of Brazil - Atlantic Forest, Pantanal, and Cerrado. Overall, a total of 5970 tick specimens representing the following nine species were collected from tapirs: Amblyomma brasiliense, Amblyomma coelebs, Amblyomma dubitatum, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma triste, Haemaphysalis juxtakochi, and Rhipicephalus microplus. Amblyomma sculptum was the most prevalent and abundant tick species in all three biomes; however, mean intensity values for A. sculptum were significantly lower in Atlantic Forest than in the Pantanal or Cerrado, and at the same time, statistically similar among tapirs from Pantanal and Cerrado. Contrastingly, mean intensity values for A. coelebs were significantly higher in the Atlantic Forest than in the other biomes. The remaining tick species were collected in lower numbers, or were exclusive for one biome, e.g., A. brasiliense and H. juxtakochi only in the Atlantic Forest. A total of 177 blood sera (123 individuals plus 54 recaptures) were collected from tapirs and tested for the presence of reactive antibodies to six Rickettsia species by immunofluorescence assay. Overall, 69% (9/13), 49% (62/126), and 66% (25/38) tapir sera from Atlantic Forest, Pantanal and Cerrado, respectively, were seroreactive to at least one Rickettsia species, with no significant difference between the three areas. Although many tapir sera reacted simultaneously to ≥2 Rickettsia species, Rickettsia parkeri elicited highest % seroprevalence and endpoint titers, and was incriminated as the possible agent involved in a homologous reaction in tapirs from the three biomes, where A. ovale was previously found infected by R. parkeri. In fact, seroconversion to R. parkeri was demonstrated in five tapirs that were captured at least twice during the study. This study demonstrated that tapirs were found to be constantly infested by several tick species in the Atlantic Forest, Pantanal and Cerrado biomes; however, the richness of tick infestations was concordant to the tick species known to be established in each biome. Under natural conditions, lowland tapirs were shown to be exposed to tick-borne spotted fever group rickettsiae.

HEALTH ASSESSMENT OF WILD LOWLAND TAPIRS (TAPIRUS TERRESTRIS) IN THE HIGHLY THREATENED CERRADO BIOME, BRAZIL.

Over 2 yr, we assessed the health of 35 lowland tapirs (Tapirus terrestris) in the Brazilian Cerrado (CE) biome, an area that is highly affected by human activities. This involved physical examinations, hematology and blood biochemistry, urinalysis, fecal parasitologic evaluation, microbial profiling of anatomic cavities and lesions, and serologic surveys for evidence of infectious agents. Research methods closely resembled those used in previous tapir health assessments in the Atlantic Forest (AF) and Pantanal (PA) biomes, allowing for a comparison among the three populations. Although not reaching statistical significance (P>0.05), tapirs from the CE exhibited poorer body and skin condition as compared to animals from the AF and PA. Furthermore, there were higher prevalences of dental problems and traumatic lesions as compared to those from the AF and PA. Eight of the 12 hematologic parameters evaluated and 17 of the 30 biochemical parameters differed significantly (P<0.05) between the tapirs from CE and those from the AF and PA. We isolated 24 different microbiologic strains from swabs of anatomic cavities and dermal lesions, of which five taxa had not previously been found in the AF or PA. We detected serum antibodies to Leptospira interrogans, bluetongue virus, and porcine parvovirus. Overall, our results suggested that tapirs from the CE exhibited more health abnormalities than tapirs in the AF and PA, possibly due to a greater exposure to environmental disturbances in the area.

Phylogenetics, patterns of genetic variation and population dynamics of Trypanosoma terrestris support both coevolution and ecological host-fitting as processes driving trypanosome evolution.

BACKGROUND: A considerable amount of evidence has favored ecological host-fitting, rather than coevolution, as the main mechanism responsible for trypanosome divergence. Nevertheless, beyond the study of human pathogenic trypanosomes, the genetic basis of host specificity among trypanosomes isolated from forest-inhabiting hosts remains largely unknown. METHODS: To test possible scenarios on ecological host-fitting and coevolution, we combined a host capture recapture strategy with parasite genetic data and studied the genetic variation, population dynamics and phylogenetic relationships of Trypanosoma terrestris, a recently described trypanosome species isolated from lowland tapirs in the Brazilian Pantanal and Atlantic Forest biomes. RESULTS: We made inferences of T. terrestris population structure at three possible sources of genetic variation: geography, tapir hosts and 'putative' vectors. We found evidence of a bottleneck affecting the contemporary patterns of parasite genetic structure, resulting in little genetic diversity and no evidence of genetic structure among hosts or biomes. Despite this, a strongly divergent haplotype was recorded at a microgeographical scale in the landscape of Nhecolândia in the Pantanal. However, although tapirs are promoting the dispersion of the parasites through the landscape, neither geographical barriers nor tapir hosts were involved in the isolation of this haplotype. Taken together, these findings suggest that either host-switching promoted by putative vectors or declining tapir population densities are influencing the current parasite population dynamics and genetic structure. Similarly, phylogenetic analyses revealed that T. terrestris is strongly linked to the evolutionary history of its perissodactyl hosts, suggesting a coevolving scenario between Perissodactyla and their trypanosomes. Additionally, T. terrestris and T. grayi are closely related, further indicating that host-switching is a common feature promoting trypanosome evolution. CONCLUSIONS: This study provides two lines of evidence, both micro- and macroevolutionary, suggesting that both host-switching by ecological fitting and coevolution are two important and non-mutually-exclusive processes driving the evolution of trypanosomes. In line with other parasite systems, our results support that even in the face of host specialization and coevolution, host-switching may be common and is an important determinant of parasite diversification.

Predicting spatiotemporal patterns of road mortality for medium-large mammals.

We modelled the spatiotemporal patterns of road mortality for seven medium-large mammals, using a roadkill dataset from Mato Grosso do Sul, Brazil (800 km of roads surveyed every two weeks, for two years). We related roadkill presence-absence along the road sections (1000 m) and across the survey dates with a collection of environmental variables, including land cover, forest cover, distance to rivers, temperature, precipitation and vegetation productivity. We further included two variables aiming to reflect the intrinsic spatial and temporal roadkill risk. Environmental variables were obtained through remote sensing and weather stations, allowing the estimate of the roadkill risk for the entire surveyed roads and survey periods. Overall, the models could explain a small fraction of the spatiotemporal patterns of roadkills (<0.23), probably due to species being habitat generalists, but still had reasonable discrimination power (AUC averaging 0.70 ±â€¯0.07). The intrinsic spatial and temporal roadkill risk were the most important variables, followed by land cover, climate and NDVI. We show that identifying spatiotemporal roadkill patterns may provide valuable information to define specific management actions focused on road sections and time periods, in complement to permanent road mitigation measures. Our approach thus offers a new insight into the understanding of road effects and how to plan and strategize monitoring and mitigation.

Health assessment of wild lowland tapir (Tapirus terrestris) populations in the Atlantic Forest and Pantanal biomes, Brazil (1996-2012).

Abstract The lowland tapir (Tapirus terrestris) is found in South America and is listed as Vulnerable to Extinction by the International Union for Conservation of Nature, Red List of Threatened Species. Health issues, particularly infectious diseases, are potential threats for the species. Health information from 65 wild tapirs from two Brazilian biomes, Atlantic Forest (AF) and Pantanal (PA), were collected during a long-term study (1996-2012). The study included physic, hematologic and biochemical evaluations, microbiologic cultures, urinalysis, and serologic analyses for antibodies against 13 infectious agents (viral and bacterial). The AF and PA tapirs were significantly different for several hematologic and biochemical parameters. Ten bacteria taxa were identified in the AF and 26 in the PA. Antibodies against five viruses were detected: Bluetongue virus, eastern equine encephalitis virus, western equine encephalitis virus, infectious bovine rhinotracheitis virus, and porcine parvovirus. A high prevalence of exposure to Leptospira interrogans (10 serovars: Autumnalis, Bratislava, Canicola, Copenhageni, Grippotyphosa, Hardjo, Hebdomadis, Icterohaemorrhagiae, Pomona, and Pyrogenes) was detected in both the AF and PA sites. A greater diversity of serovars and higher antibody titers were found in the PA. Statistically significant differences between sites were found for L. interrogans, equine encephalitis virus, and porcine parvovirus. Based on physical evaluations, both AF and PA populations were healthy. The differences in the overall health profile of the AF and PA tapir populations appear to be associated with environmental factors and infectious diseases ecology. The extensive datasets on hematology, biochemistry, urinalysis, and microbiology results from this paper can be used as reference values for wild tapirs.

Tapir health and conservation medicine.

Tapirs have unique nutritional needs, as well as anatomical, physiological, behavioral and ecological adaptations that must be considered when managing their health, both in the wild and in captivity. Information about how tapirs live in their natural habitats can provide crucial knowledge to prevent many of the health problems found in captivity such as infectious and parasitic diseases, reproductive issues and nutritional and behavioral disorders. Likewise, proper management in captivity can significantly contribute to in situ conservation programs. Conservation medicine is a science created to address the global health crisis that jeopardizes biodiversity causing imbalances among ecosystem, human, animal and vegetal health. In this context, common threats to tapir health and conservation, such as isolated and small populations surrounded by human activity, chemical pollution, domestic animals and their pathogenic agents, need to be better understood. This manuscript provides information about the health of tapirs both in captivity and in the wild and aims to encourage tapir conservationists worldwide to gather information about pathogen and disease dynamics and manifestation, as well as implications for tapir conservation.

Distribution, habitat and adaptability of the genus Tapirus.

In this manuscript, as a starting point, the ancient and current distribution of the genus Tapirus are summarized, from its origins, apparently in Europe, to current ranges. Subsequently, original and current tapir habitats are described, as well as changes in ancient habitats. As the manuscript goes on, we examine the ways in which tapir species interact with their habitats and the main aspects of habitat use, spatial ecology and adaptability. Having reviewed the historic and current distribution of tapirs, as well as their use and selection of habitats, we introduce the concept of adaptability, considering that some of the tapir physiological characteristics and behavioral strategies can reduce the negative impact of habitat alteration and climate change. Finally, we provide recommendations for future research priorities. The conservation community is still missing important pieces of information for the effective conservation of tapirs and their remaining habitats in Central and South America and Southeast Asia. Reconstructing how tapir species reached their current distribution ranges, interpreting how they interact with their habitats and gathering information regarding the strategies they use to cope with habitat changes will increase our understanding about these animals and contribute to the development of conservation strategies.

Population viability analysis: using a modeling tool to assess the viability of tapir populations in fragmented landscapes.

A population viability analysis (PVA) was conducted of the lowland tapir populations in the Atlantic Forest of the Pontal do Paranapanema region, Brazil, including Morro do Diabo State Park (MDSP) and surrounding forest fragments. Results from the model projected that the population of 126 tapirs in MDSP is likely to persist over the next 100 years; however, 200 tapirs would be required to maintain a viable population. Sensitivity analysis showed that sub-adult mortality and adult mortality have the strongest influence on the dynamics of lowland tapir populations. High road-kill has a major impact on the MDSP tapir population and can lead to population extinction. Metapopulation modeling showed that dispersal of tapirs from MDSP to the surrounding fragments can be detrimental to the overall metapopulation, as fragments act as sinks. Nevertheless, the model showed that under certain conditions the maintenance of the metapopulation dynamics might be determinant for the persistence of tapirs in the region, particularly in the smaller fragments. The establishment of corridors connecting MDSP to the forest fragments models resulted in an increase in the stochastic growth rate, making tapirs more resilient to threats and catastrophes, but only if rates of mortality were not increased when using corridors. The PVA showed that the conservation of tapirs in the Pontal region depends on: the effective protection of MDSP; maintenance and, whenever possible, enhancement of the functional connectivity of the landscape, reducing mortality during dispersal and threats in the unprotected forest fragments; and neutralization of all threats affecting tapirs in the smaller forest fragments.