Clinical and diagnostic evaluation of a wild ocelot (Leopardus pardalis) specimen.

Background: Monitoring the health of wild animals under the principles of one health contributes to the prevention of diseases and the preservation of human and animal health, thus contributing to the conservation of species. Aim: The current study describes the clinical and paraclinical status of an ocelot (Leopardus pardalis) captured in Buriticá, Antioquia, Colombia, for research purposes with the aim of contributing to the construction of animal health reference values in the wild, considering the scarcity of published data for the country on capture, management, and paraclinical parameters related to this species. Methods: For this, hematological parameters, blood chemistry, urine cytochemical analyses, and coprological examinations were carried out. Results: The hematological values of the captured individual do not show relevant differences concerning those reported in the literature in both captive and free specimens. However, differences were identified between the reference blood chemistry and urine cytochemical values between reports of animals in captivity and the wild, revealing the need to develop reference standards for animals in the wild that guarantee adequate management of these species and favor their conservation. A possible picture of renal failure and multiple parasitic infections of epidemiological importance was found. Conclusion: This study reports for the first time a urine infection by Capillaria sp. and an infection by Dicrocoelium spp. in fecal matter for the species.

Exposing illegal hunting and wildlife depletion in the world's largest tropical country through social media data.

Globally, illegal sport hunting can threaten prey populations when unregulated. Due to its covert nature, illegal sport hunting poses challenges for data collection, hindering efforts to understand the full extent of its impacts. We gathered social media data to analyze patterns of illegal sport hunting and wildlife depletion across Brazil. We collected data for 2 years (2018-2020) across 5 Facebook groups containing posts depicting pictures of illegal sport hunting events of native fauna. We described and mapped these hunting events by detailing the number of hunters involved, the number of species, the mean body mass of individuals, and the number and biomass of individuals hunted per unit area, stratified by Brazilian biome. We also examined the effects of defaunation on hunting yield and composition via regression models, rank-abundance curves, and spatial interpolation. We detected 2046 illegal sport hunting posts portraying the hunting of 4658 animals (∼29 t of undressed meat) across all 27 states and 6 natural biomes of Brazil. Of 157 native species targeted by hunters, 19 are currently threatened with extinction. We estimated that 1414 hunters extracted 3251 kg/million km2. Some areas exhibited more pronounced wildlife depletion, in particular the Atlantic Forest and Caatinga biomes. In these areas, there was a shift from large mammals and reptiles to small birds as the main targeted taxa, and biomass extracted per hunting event and mean body mass across all taxonomic groups were lower than in other areas. Our results highlight that illegal sport hunting adds to the pressures of subsistence hunting and the wild meat trade on Brazil's wildlife populations. Enhanced surveillance efforts are needed to reduce illegal sport hunting levels and to develop well-managed sustainable sport hunting programs. These can support wildlife conservation and offer incentives for local communities to oversee designated sport hunting areas.

Exposición de la caza ilegal y la reducción de fauna en el país tropical más grande del mundo por medio de datos de las redes sociales Resumen En todo el mundo, la caza recreativa ilegal puede amenazar a las poblaciones de presas cuando no está regulada. Debido a su naturaleza encubierta, la caza recreativa ilegal plantea dificultades para la recopilación de datos, lo que dificulta la comprensión de su impacto. Recopilamos datos de redes sociales para analizar los patrones de caza recreativa ilegal y agotamiento de la vida silvestre en todo Brasil. Recopilamos datos durante 2 años (2018­2020) a través de cinco grupos de Facebook que contenían publicaciones que mostraban imágenes de eventos de caza recreativa ilegal de fauna nativa. Describimos y mapeamos estos eventos de caza detallando el número de cazadores involucrados, el número de especies, la masa corporal media de los individuos y el número y la biomasa de los individuos cazados por unidad de área, estratificados por bioma brasileño. También examinamos los efectos de la deforestación en el rendimiento y la composición de la caza mediante modelos de regresión, curvas de abundancia e interpolación espacial. Detectamos 2,046 puestos de caza recreativa ilegal que mostraban la caza de 4,658 animales (∼29 t de carne sin desollar) en los 27 estados y 6 biomas naturales de Brasil. De las 157 especies autóctonas objetivo de los cazadores, 18 están actualmente en peligro de extinción. Se calcula que 1,414 cazadores extrajeron 3,251 kg/millón de km2. Algunas zonas mostraron una defaunación más pronunciada, en particular los biomas de la Mata Atlántica y la Caatinga. En estas áreas, se produjo un cambio de grandes mamíferos y reptiles a pequeñas aves como principales taxones objetivo, y la biomasa extraída por evento de caza y la masa corporal media en todos los grupos taxonómicos fueron menores que en otras áreas. Nuestros resultados ponen de manifiesto que la caza recreativa ilegal se suma a las presiones de la caza de subsistencia y el comercio de carne salvaje sobre las poblaciones de fauna de Brasil. Es necesario intensificar los esfuerzos de vigilancia para reducir los niveles de caza recreativa ilegal y desarrollar programas de caza recreativa sostenibles y bien gestionados. Estos programas pueden contribuir a la conservación de la fauna y ofrecer incentivos a las comunidades locales para que supervisen las zonas designadas para la caza recreativa.

Detection of Brucella S19 Vaccine Strain DNA in Domestic and Wild Ungulates from Brazilian Pantanal.

The Pantanal region, the largest floodplain in the world, has a huge biodiversity and is an important livestock center. Bovine brucellosis has been reported in the region over the last three decades, posing implications for cattle industry as well as for the maintenance of biodiversity. We aimed to investigate the presence of B. abortus S19 vaccine strain DNA in unvaccinated domestic and wild ungulates from the Brazilian Pantanal. Fifty-two heifers, 63 ovine, 24 domestic pigs, 28 feral pigs, and three Pampas deer were sampled. Brucella spp. was detected through bcsp31 PCR of blood samples in 45.3% (77/170) of the sampled animals, of which 36.4% (28/77) showed positivity in ery PCR corresponding to B. abortus S19 strain. Feral pigs presented the highest occurrence of positive samples in bcsp31 PCR (75%), followed by ovine (47.6%), domestic pigs (41.7%), and unvaccinated heifers (30.8%). We did not observe positivity in Pampas deer. Our results strongly suggest that vaccination against bovine brucellosis may promote spill-over of B. abortus S19 strain in the Pantanal region. Moreover, our data indicate that wild strains of Brucella circulates in the Pantanal Biome.

Tick-borne Apicomplexa in wildlife and ticks of French Guiana.

Tick-borne Apicomplexa encompass a group of parasites responsible for significant medical and veterinary diseases, including babesiosis, theileriosis, and hepatozoonosis. In this study, we investigated the presence and diversity of tick-borne Apicomplexa in wildlife and ticks inhabiting the Amazon rainforests of French Guiana. To this end, we conducted molecular screening and typing using 18S rRNA sequences on a collection of 1161 specimens belonging to 71 species, including 44 species of wild mammals, five species of passerines, and 22 species of ticks. We characterized eight genovariants of Babesia, Theileria, Hemolivia, and Hepatozoon parasites, some matching known species, while others suggested potential novel species. These parasites were detected in wild mammals, including opossums, sloths, armadillos, porcupines, margays, greater grisons, and ticks, but not in passerines. Finally, similarities with surveys conducted in Brazil highlight the specific sylvatic transmission cycles of South American tick-borne Apicomplexa.

Title: Apicomplexes transmis par les tiques chez la faune sauvage et les tiques de Guyane française. Abstract: Les Apicomplexes transmis par les tiques englobent un groupe de parasites responsables de maladies médicales et vétérinaires importantes, notamment la babésiose, la theilériose et l'hépatozoonose. Dans cette étude, nous avons étudié la présence et la diversité des Apicomplexes transmis par les tiques dans la faune sauvage et les tiques habitant les forêts tropicales amazoniennes de Guyane française. À cette fin, nous avons effectué un criblage moléculaire et un typage à l'aide de séquences d'ARNr 18S sur une collection de 1 161 spécimens appartenant à 71 espèces, dont 44 espèces de mammifères sauvages, cinq espèces de passereaux et 22 espèces de tiques. Nous avons caractérisé huit génovariants des parasites Babesia, Theileria, Hemolivia et Hepatozoon, certains correspondant à des espèces connues tandis que d'autres suggéraient de nouvelles espèces potentielles. Ces parasites ont été détectés chez des mammifères sauvages, dont des opossums, des paresseux, des tatous, des porcs-épics, des margays, des grisons et des tiques, mais pas chez des passereaux. Enfin, des similitudes avec des enquêtes menées au Brésil mettent en évidence les cycles de transmission sylvatiques spécifiques des Apicomplexa transmis par les tiques d'Amérique du Sud.

Infection of South American coatis (Nasua nasua) with highly pathogenic avian influenza H5N1 virus displaying mammalian adaptive mutations.

Deadly outbreaks among poultry, wild birds, and carnivorous mammals by the highly pathogenic H5N1 virus of the clade 2.3.4.4b have been reported in South America. The increasing virus incidence in various mammal species poses a severe zoonotic and pandemic threat. In Uruguay, the clade 2.3.4.4b viruses were first detected in February 2023, affecting wild birds and backyard poultry. Three months after the first reported case in Uruguay, the disease affected a population of 23 coatis (Nasua) in an ecological park. Most animals became infected, likely directly or indirectly from wild birds in the park, and experienced sudden death. Five animals from the colony survived, and four of them developed antibodies. The genomes of the H5N1 strains infecting coatis belonged to the B3.2 genotype of the clade 2.3.4.4b. Genomes from coatis were closely associated with those infecting backyard poultry, but transmission likely occurred through wild birds. Notable, two genomes have a 627K substitution in the RNA polymerase PB2 subunit, a hallmark amino acid linked to mammalian adaptation. Our findings support the ability of the avian influenza virus of the 2.3.4.4b clade to infect and transmit among terrestrial mammals with high pathogenicity and undergo rapid adaptive changes. It also highlights the coatis' ability to develop immunity and naturally clear the infection.

Serologic Survey of Brucella spp. in Culled Invasive Alien Mammals from El Palmar National Park, Argentina, and in Exposed Consumers.

Brucellosis is a worldwide zoonotic disease caused by Brucella spp. and transmitted from domestic and wild animals to humans. Brucellosis causes important economic losses in livestock, has a significant impact on public health, and may affect the health of wildlife. Hunting and consumption of meat from culled wildlife constitute a risk for Brucella spp. infection in humans and hunting dogs. In El Palmar National Park (EPNP), Argentina, the invasive alien mammals wild boar (Sus scrofa) and axis deer (Axis axis) are controlled, slaughtered in situ, and consumed by hunters, with meat trimmings and offal often fed to dogs. In this study, we evaluated but did not detect anti-Brucella antibodies in wild boar (n=95) and axis deer (n=238) from EPNP or in game consumers, dogs (n=39) and humans (n=61). These results suggest a lack of exposure to Brucella spp. at this site during the study period. Despite negative findings in the sampled location, One Health surveillance across multiple species contributes to our understanding of pathogen dynamics and enables targeted interventions to minimize health risks.

Rabbit Hemorrhagic Disease Virus in Mexico in 2020-2021: Risk Areas and Climatic Distribution.

Mexico is home to 14 species of lagomorphs, 6 of which are endemic. Studies on diseases affecting native lagomorphs are scarce, and in most cases, the impact on their populations remains largely unknown. Rabbit hemorrhagic disease virus (RHDV), especially the RHDV2 variant, causes a serious and extremely contagious disease, resulting in high mortality rates and major declines in wild lagomorph populations. The objectives of this study were to identify disease hotspots and critical biodiversity regions in Mexico through the combined use of disease information and lagomorph distribution maps and to determine the areas of greatest concern. In total, 19 states of Mexico recorded RHDV2 from April 2020 to August 2021, and 12 of them reported the wild species Sylvilagus audubonii, Lepus californicus, and unidentified Leporidae species. The distribution of RHDV2 in Mexico can be closely predicted from climatic variables. RHDV2 hotspots are located in the central-southern area of the Mexican Highlands and the Trans-Mexican Volcanic Belt, where the virus affects multiple species. This knowledge is essential for proposing specific actions to manage and preserve lagomorph populations at risk and address these issues as soon as possible.

The use of the micronucleus test and comet assay in wild rodents: a historical review and future perspectives.

Rodents are considered good models for investigating genotoxic damage and mutagenic alterations caused by xenobiotic agents, due to their occupation of a wide variety of habitats. However, relatively few in situ studies have focused on DNA damage in wild rodents associated with environmental exposure. In this review, we investigate trends in the application of the micronucleus test and comet assay in in situ studies of wild rodents. A total of 33 papers were identified, distributed across 14 different countries. Brazil and Spain had the most published studies (six each), followed by Bulgaria (n = 5), Mexico (n = 4) and Italy (n = 3). Only 24 of the 2,652 recognized rodent species have been the subject of in situ studies, which have most frequently focus on species of the genus Mus. The protocols used for the micronucleus test and comet assay varied widely, although blood and bone marrow were the primary types of tissue used. Given the paucity of studies on wild rodents, we recommend further research, particularly focusing on the use of this group as bioindicators of environmental quality and the standardization of protocols.

Hemoplasmas in wild rodents and marsupials from the Caatinga Biome, Brazil.

A total of 231 blood samples from wild mammals belonging to the orders Rodentia (n = 142) and Didelphimorphia (n = 89) were screened by real-time PCR assay (qPCR), being six Rhipidomys sp., 118 Thrichomys laurentius, nine Rattus rattus, four Kerodon rupestris, five Necromys lasiurus, 42 Didelphis albiventris and 47 Monodelphis domestica. Results using qPCR showed that 32 of the total 231 (13.85 %) samples were positive for hemoplasma sequences of the 16S rRNA gene. Sequences from two D. albiventris showed 99.77-99.89 % identity with 'Candidatus Mycoplasma haemoalbiventris' and 99.09 % with 'Candidatus Mycoplasma haemodidelphidis', respectively. Furthermore, one M. domestica and five T. laurentius showed 99.72-99.77 % identity with Mycoplasma sp., and one K. rupestris showed 98.13 % identity with 'Candidatus Mycoplasma haematohydrochaerus'; and from two Rattus rattus showed 99.65-99.89 % identity with Mycoplasma sp. and 'Candidatus Mycoplasma haemomuris'. The 23S rRNA gene sequences obtained from the two D. albiventris showed 100 % identity with 'Ca. M. haemoalbiventris' whereas the sequences from the R. rattus showed only 85.31 % identity with 'Candidatus Mycoplasma haematohydrochaerus'. Two T. laurentius and one K. rupestris showed 84.66-92.97 % identity with 'Candidatus Mycoplasma haemosphiggurus'. Based on phylogenetic and Neighbor-Net network analyses of the 16S and 23S rRNA genes, potential novel species are described. In addition, 'Ca. M. haemoalbiventris' was detected in Didelphis albiventris, and Mycoplasma sp. was detected in Rattus sp. rodents from the Caatinga biome, Brazil.

Zoonotic Mansonella ozzardi in Raccoons, Costa Rica, 2019-2022.

Mansonella ozzardi, a filarioid parasite, causes human mansonellosis in the Americas. We identified raccoons (Procyon lotor) as wildlife reservoirs of M. ozzardi in Costa Rica. Noting the sympatry of free-ranging raccoons and humans, we conclude that mansonellosis is a considerable public health risk in the region.

Records of the Invasive Cattle Tick Rhipicephalus microplus on Wild Hosts and of a Neotropical Tick, Amblyomma dubitatum, on Invasive Alien Mammals, Southern Brazil.

Invasive species threaten wildlife. We detected invasive ticks on wildlife and indigenous ticks on invasive mammals in southern Brazil. The invasive cattle tick Rhipicephalus microplus was found on 12 native mammals and one invasive exotic (Axis axis). The Neotropical tick Amblyomma dubitatum was found on three invasive alien mammal species.

Investigation of Rabies virus in wild mammals of the atlantic forest in Rio de Janeiro, Brazil.

With the successful control of rabies transmitted by dogs in Brazil, wild animals have played a relevant epidemiological role in the transmission of rabies virus (RABV). Bats, non-human primates and wild canines are the main wild animals that transmit RABV in the country. It is worth highlighting the possibility of synanthropic action of these species, when they become adapted to urban areas, causing infections in domestic animals and eventually in humans. This work aimed to evaluate the circulation of RABV in the Pedra Branca Forest, an Atlantic Forest area, located in the state of Rio de Janeiro, Southeast Brazil. Saliva and blood samples were obtained from 60 individuals of eight species of bats, captured with mist nets, and 13 individuals of callitrichid primates, captured with tomahawk traps. Saliva samples were subjected to Reverse Transcription Polymerase Chain Reaction (RT-PCR), targeting the RABV N gene, with all samples being negative. Blood samples of all animals were submitted to the Rapid Fluorescent Focus Inhibition Test (RFFIT) to detect neutralizing antibodies (Ab) for RABV. Six bat samples (8%) were seropositive for RABV with antibody titers greater than or equal to 0.1 IU/mL. The detection of Ab but not viral RNA indicates exposure rather than current RABV transmission in the analyzed populations. The results presented here reinforce the importance of serological studies in wildlife to access RABV circulation in a region.

Evaluating climate-dependent distribution of orthohantaviruses with monitoring wild rodents: One Health Perspective†.

Orthohantaviruses, cause hemorrhagic fever with renal syndrome, nephropathia epidemica, and hantavirus pulmonary syndrome, are major public health problems all over the world. Wild rodent surveillance for orthohantaviruses is of great importance for the preparedness against these human infections and the prediction of possible outbreak regions. Thus, we aimed to screen orthohantaviruses in wild rodents in Southern Anatolia, where the area has some of the glacial period refugia in the Mediterranean Basin, and interpret their current epidemiology with climatic biovariables in comparison with previously positive regions.We trapped muroid rodents between 2015 and 2017, and screened for orthohantaviruses. Then, we evaluated the relationship between orthohantavirus infections and bioclimatic variables. In spite of the long-term and seasonal sampling, we found no evidence for Orthohantavirus infections. The probable absence of orthohantaviruses in the sampling area was further evaluated from the climatic perspective, and results led us suggest that Orthohantavirus epidemiology might be relatively dependent on precipitation levels in driest and warmest quarters, and temperature fluctuations.These initial data might provide necessary perspective on wild rodent surveillance for orthohantaviruses in other regions, and help to collect lacking data for a such habitat suitability study in a bigger scale in the future.

Re-domestication of feral pigs in Martinique: first results of growing-finishing trials to test meat consumption and consumers' perceptions.

In Martinique, pig production satisfies less than 20% of demand for pork, with traditional pig farming and wild pig hunting covering only a small proportion. This study has three parts: (i) for the first time, it analyses the performance of domesticated descendants of Creole feral pigs of Martinique based on a 29-day finishing test on two farms (A vs. B) with the same fibre-rich diet on a total of 40 pigs; (ii) it evaluates consumers' tastes based on a sensorial test by an untrained panel of 61 consumers who tasted pork prepared as a fricassee from either farm A or farm B; and (iii) evaluates willingness to pay (WTP) for Creole pork products and of the design of a future niche market. The average daily gain (ADG) of the finishing pigs was higher on farm A than on farm B (256 vs. 100 g/d, P < 0.001) resulting in a higher hot carcass weight on farm A than farm B (41.3 vs. 33.5 kg, P < 0.01), and better hot carcass yield on farm B than farm A (74.0 vs 68.8 %, P < 0.01). In the sensorial test, the consumers gave a higher score to the meat originating from the youngest pigs (Farm B), especially scores for tenderness and juiciness (+0.94 and +0.55 points, P < 0.05 and P=0.10, respectively). The 61 respondents were willing to pay more for processed products (sausage, pâté, ham) than for fresh meat. They considered that Creole pork has a better taste and is of better quality than industrial meats originating from mainstream genotypes. According to the majority of respondents, Creole pork should be rich in intra-muscular fat (100% of respondents) and low in inter-muscular fat (60% of respondents). Based on the survey, the main desirable future for Martinique's Creole pig production correspond to a low-carbon system with feed based on local resources, with on-farm slaughtering and short-distance sales (direct-to-consumer farm gate sale, sale at the butcher's or at the pig cooperative).

Detection of rabies virus in Callithrix penicillata (Geoffroy, 1812) in Montes Claros, Minas Gerais State, Brazil.

This report describes the occurrence of the rabies virus in two species of wild animals in the urban area of Montes Claros (MOC), Minas Gerais State, Brazil, in May 2023. The virus has been detected in frugivorous chiropterans (Artibeus sp) and marmosets (Callithrix penicillata). This is the first notified case of the rabies virus in the species C. penicillata in the urban area of MOC. Our findings show that the rabies virus is circulating in the urban area of MOC; therefore, permanent preventive measures must be adopted to avoid infection of other animals and humans.

Risk for Waterborne Transmission and Environmental Persistence of Avian Influenza Virus in a Wildlife/Domestic Interface in Mexico.

Aquatic habitats provide a bridge for influenza transmission among wild and domestic species. However, water sources pose highly variable physicochemical and ecological characteristics that affect avian influenza virus (AIV) stability. Therefore, the risk of survival or transmissibility of AIV in the environment is quite variable and has been understudied. In this study, we determine the risk of waterborne transmission and environmental persistence of AIV in a wild/domestic bird interface in the Central Mexico plateau (North America) during the winter season using a multi-criteria decision analysis (MCDA). A total of 13 eco-epidemiological factors were selected from public-access databases to develop the risk assessment. The MCDA showed that the Atarasquillo wetland presents a higher persistence risk in January. Likewise, most of the backyard poultry farms at this wild-domestic interface present a high persistence risk (50%). Our results suggest that drinking water may represent a more enabling environment for AIV persistence in contrast with wastewater. Moreover, almost all backyard poultry farms evidence a moderate or high risk of waterborne transmission especially farms close to water bodies. The wildlife/domestic bird interface on the Atarasquillo wetland holds eco-epidemiological factors such as the presence of farms in flood-prone areas, the poultry access to outdoor water, and the use of drinking-water troughs among multiple animal species that may enhance waterborne transmission of AIV. These findings highlight the relevance of understanding the influence of multiple factors on AIV ecology for early intervention and long-term control strategies.

Safety and Immunogenicity of Morbillivirus canis Vaccines for Domestic and Wild Animals: A Scoping Review.

Morbillivirus canis (canine distemper virus (CDV)) is recognized as a multihost pathogen responsible for a transmissible disease affecting both domestic and wild animals. A considerable portion of wildlife populations remain unvaccinated due to a lack of safety and immunogenicity data on existing vaccines for the prevention of CDV infection in these species. This review aimed to assess the current state of CDV vaccination research for both domestic and wild animals and to explore novel vaccine candidates through in vivo studies. It also sought to synthesize the scattered information from the extensive scientific literature on CDV vaccine research, identify key researchers in the field, and highlight areas where research on CDV vaccination is lacking. A scoping review was conducted across four databases following the PRISMA-ScR protocol, with information analyzed using absolute and relative frequencies and 95% confidence intervals (CIs) for study number proportions. Among the 2321 articles retrieved, 68 met the inclusion criteria and focused on CDV vaccines in various animal species, such as dogs, ferrets, minks, and mice. Most of the scientific community involved in this research was in the USA, Canada, France, and Denmark. Various vaccine types, including MLV CDV, recombinant virus, DNA plasmids, inactivated CDV, and MLV measles virus (MeV), were identified, along with diverse immunization routes and schedules employed in experimental and commercial vaccines. Safety and efficacy data were summarized. Notably, 37 studies reported postimmunization CDV challenge, primarily in dogs, revealing the survival rates of vaccinated animals. In summary, CDV vaccines generally demonstrate an acceptable safety profile in dogs and show promise as a means of controlling CDV. However, significant gaps in vaccine research persist, particularly concerning wildlife reservoirs, indicating the need for further investigation.

Guidelines and considerations for capturing and collaring wild primates: a case study with capuchin monkeys (Sapajus nigritus cucullatus).

Many aspects of the ecology, evolution and social behavior of wild-living primates remain un-explored and require further investigation. While long-term field studies are crucial for addressing conservation concerns for many primates' species, acquiring the necessary data is often challenging, often due to difficulties in locating study groups. Radio-telemetry has significantly facilitated the study of primates and other animals living in tropical forests. However, there are important practical challenges in the process of capturing and releasing animals after placement of telemetry collars. In this study, we report guidelines and considerations for capturing and collaring wild capuchin monkeys, Sapajus nigritus cucullatus, in the Atlantic Forest of Argentina. Our ultimate goal is to contribute to making captures safer, preventing harm and stress to animals when using radio-telemetry in monitoring strategies for conservation of this primate species. These methods can be useful for researchers using field capture and radio-telemetry for monitoring groups or populations of wild primates, specifically wild Sapajus.

Absence of Leishmania spp. DNA in road-killed wild mammals in Southern Brazil.

Leishmaniasis are neglected diseases transmitted by vectors that affect domestic and wild animals, including humans. Due to its incidence and lethality, this zoonosis is a worrying public health problem, making it essential to identify all links in the transmission chain. Infection of wild mammals by Leishmania spp. remains poorly understood, especially in southern Brazil. Therefore, the objective was to research, using the PCR technique, the presence of Leishmania spp. DNA in road-killed wild mammals in Southern Brazil. Carcasses of 96 animals were collected from highways in the Pelotas microregion, Rio Grande do Sul, southern Brazil and subjected to necropsies. Tissue fragments (spleen, skin, liver, kidney, heart, lung, lymph nodes, bone marrow and blood) were collected and genomic DNA was extracted. PCR protocols targeting the ITS1, kDNA and 18S genes were tested. We found no evidence of Leishmania spp. circulation in the studied population. However, epidemiological studies like this one are of great relevance, as they allow monitoring of the occurrence of pathogens and help identify possible risk areas. As these animals act as epidemiological markers for the presence of the microorganism, studies must be carried out continuously to understand whether there are sources of infection in the region.