Rabies virus circulation in a highly diverse bat assemblage from a high-risk area for zoonoses outbreaks in the Brazilian Amazon.

Bats are the second most diverse order of mammals and play a central role in ecosystem dynamics. They are also important reservoirs of potentially zoonotic microorganisms, of which rabies virus is the most lethal among the bat-transmitted zoonotic pathogens. Importantly, recent outbreaks of human rabies have been reported from the Brazilian Amazon. Here we present a survey of bat species and rabies virus (RABV) circulation in a bat assemblage in the Marajó region, northern Brazil. Using data from mist-net captures and bioacoustic sampling, 56 bat species were recorded along the Jacundá River basin over a 10-day expedition in November 2022. For the investigation of RABV, we used the direct fluorescent antibody test (DFAT) and the rapid fluorescent focus inhibition test (RFFIT). In total, 159 bat individuals from 22 species were investigated for RABV. Five adults of the common vampire bat, Desmodus rotundus, showed RABV-specific antibodies in serum samples. Additionally, we report on local residents with injuries caused by D. rotundus bites and the occurrence of colonies of non-hematophagous bats from different species roosting inside human residences. This scenario raises concerns about the risks of new cases of human rabies and other zoonotic diseases associated with bats in the region and highlights the need for epidemiological surveillance and mitigation measures to prevent outbreaks of emerging infectious diseases.

Corrigendum to "Trypanosoma spp. Neobats: Insights about those poorly known trypanosomatids" [Int. J. Parasitol. Parasites Wildl. 16 (2021) 145-152].

[This corrects the article DOI: 10.1016/j.ijppaw.2021.09.003.].

Old Methods, New Insights: Reviewing Concepts on the Ecology of Trypanosomatids and Bodo sp. by Improving Conventional Diagnostic Tools.

Mixed infections by different Trypanosoma species or genotypes are a common and puzzling phenomenon. Therefore, it is critical to refine the diagnostic techniques and to understand to what extent these methods detect trypanosomes. We aimed to develop an accessible strategy to enhance the sensitivity of the hemoculture, as well as to understand the limitations of the hemoculture and the blood clot as a source of parasitic DNA. We investigated trypanosomatid infections in 472 bats by molecular characterization (18S rDNA gene) of the DNA obtained from the blood clot and, innovatively, from three hemoculture sample types: the amplified flagellates ("isolate"), the pellet of the culture harvested in its very initial growth stage ("first aliquot"), and the pellet of non-grown cultures with failure of amplification ("sediment"). We compared (a) the characterization of the flagellates obtained by first aliquots and isolates; and (b) the performance of the hemoculture and blood clot for trypanosomatid detection. We observed: (i) a putative new species of Bodo in Artibeus lituratus; (ii) the potential of Trypanosoma cruzi selection in the hemoculture; (iii) that the first aliquots and sediments overcome the selective pressure of the hemoculture; and (iv) that the blood clot technique performs better than the hemoculture. However, combining these methods enhances the detection of single and mixed infections.

Taxonomic re-evaluation of New World Eptesicus and Histiotus (Chiroptera: Vespertilionidae), with the description of a new genus

Eptesicus Rafinesque, 1820 is widely distributed in the Old and New World (26 species), and Histiotus Gervais, 1856 is a South American endemic (11 species). Molecular phylogenies have recovered Eptesicus (sensu lato) as polyphyletic, with New World Eptesicus and the sister genus Histiotus in a paraphyletic American clade sister to Old World Eptesicus. Based on these phylogenetic reconstructions, authors have treated Histiotus as either a subgenus of Eptesicus or restricted Eptesicus to the New World species, treating Histiotus as a full genus, and using the name Cnephaeus Kaup, 1829 at the generic rank to comprise Old World Eptesicus. Based on recently published molecular studies, and on novel qualitative and quantitative morphological comparisons of representatives of Histiotus and New and Old World Eptesicus, we provide evidence for restricting the name Eptesicus to the species E. fuscus (Palisot de Beauvois, 1796) and E. guadeloupensis Genoways & Baker, 1975, allocating the remaining New World species under a new genus, keeping Histiotus as a full genus, and raising Cnephaeus to generic rank to comprise all Old World taxa currently under Eptesicus. This arrangement resolves the paraphyly of New World Eptesicus, and promotes taxonomic stability for Histiotus, which is a well-established genus of easily recognizable Neotropical bats and treated separate from Eptesicus by most authorities.

Multiple colors in anteaters: review and description of chromatic disorders in Tamandua (Xenarthra: Pilosa) with reports of new and rare coat colorations

Tamandua Gray, 1825 contains two species of anteaters: Tamandua mexicana (Saussure, 1860), which occurs in southern Mexico, throughout Central America and south to the west of the Andes in South America, and Tamandua tetradactyla (Linnaeus, 1758), widely distributed in South America, from Venezuela to northern Argentina, as well as Trinidad and Tobago. Historically, different coloration patterns have been attributed to the coat of Tamandua, with T. mexicana typically showing a partial or complete dark "vest" on the dorsal portion of the body, whereas T. tetradactyla can show a complete, partial, or absent vest and a coat varying from golden, pale yellow and pale gray to brown or black. These different coat colors are sometimes treated as inherent characteristics of subspecies of Tamandua. Here, we review color variation in Tamandua and describe six chromatic disorders for the genus: albinism, leucism, "brown" variation, melanism (partial and total), xanthochromism (partial and total) and, for the first time for anteaters, erythrism. In addition, we detail the morphological expression and geographic distribution of these anomalies in populations of Tamandua from Central and South America.(AU)

Population genetic structure and morphological diversity of Cruzia tentaculata (Nematoda: Ascaridida), a parasite of marsupials (Didelphinae), along the Atlantic Forest on the eastern coast of South America.

Cruzia tentaculata is a helminth parasite of marsupials and has a wide geographic distribution from Mexico to Argentina. The aim of this study was to analyse the genetic population structure of this nematode along the Atlantic Forest biome. Cruzia tentaculata specimens were recovered from Didelphis aurita, Didelphis albiventris and Philander quica in 9 localities. Morphological and morphometric data were investigated for phenotypic diversity among localities and hosts using multivariate discriminant analysis of principal components. Phylogenetic relationships of C. tentaculata were determined using maximum likelihood and Bayesian inference. The population structure was analysed by fixation indices, molecular variance analysis, Tajima's D and Fu's Fs neutrality tests, Mantel tests and Bayesian clustering analysis. A higher significant morphometric difference for males was observed between localities. In the haplogroup networks, 2 groups were recovered, separating locations from the north and from the south/southeast. The morphometric variation in C. tentaculata between different localities was compatible with this north and southeast/south pattern, suggesting adaptation to different ecological conditions. Population genetic analyses suggested a pattern of evolutionary processes driven by Pleistocene glacial refugia in the northeast and southeast of the Atlantic Forest based on the distribution of genetic diversity.

Survey of medium- and large-sized mammals in Atlantic Forest remnants of Conceição dos Ouros, Minas Gerais, Brazil.

Conceição dos Ouros is located in the Mantiqueira mountain range (elevation: 831‒1443 m a.s.l.), State of Minas Gerais, southeast Brazil. The largest two Atlantic Forest remnants of the Municipality of Conceição dos Ouros total more than 2,000 ha and the main vegetation type is seasonal semi-deciduous forest, isolated by a matrix of agricultural fields and pasture. The Municipality does not have any protected areas and is located in a highly fragmented region, albeit considered of special importance for the conservation of terrestrial vertebrates in the State of Minas Gerais. Due to a cooperation with the Municipality of Conceição dos Ouros to carry out a survey of the local biodiversity, in this study we present the results of the survey of medium- and large-sized terrestrial mammals from the two forest remnants in the region. Sampling was performed from July 2019 to August 2021 and comprised a camera trap survey, active searching including direct (e.g. carcass sightings) and indirect (e.g. footprints and faeces) evidence of species presence and interviews with residents. Twenty-nine native and two non-native species were documented. Ten species are in some category of threat of extinction at regional, national or global levels. This is the first survey of the terrestrial mammal fauna in the area of Conceição dos Ouros and results could be helpful in designing conservation strategies at the local scale.

Systematic review of Myotis (Chiroptera, Vespertilionidae) from Chile based on molecular, morphological, and bioacoustic data.

Myotis is the most diverse genus of bats in the world, with more than 30 species recognized in the Neotropics. However, many of these species represent cryptic complexes and are evidence of the existence of hidden diversity in several regions. Using an integrative approach based on molecular, morphological, and bioacoustic data, we performed a systematic review of Myotis species from Chile. Phylogenetic inference using cytochrome-b indicated the existence of three monophyletic lineages, and qualitative and quantitative morphological analyses supported these lineages as distinct and morphologically diagnosable taxa. Analysis of discriminant functions using parameters of echolocation calls also indicates the existence of three distinct bioacoustic clusters. Thus, all lines of evidence congruently indicate the existence of three distinct taxa. As a result, we recognize Myotis arescens as a valid and distinct species and define its taxonomic limits from the other species from Chile, Myotis atacamensis and Myotis chiloensis.

Medium- and large-sized mammals from Estação Biológica Fiocruz Mata Atlântica, Rio de Janeiro, south-eastern Brazil.

The Pedra Branca Forest is in a highly urbanised region of the central portion of Rio de Janeiro City and comprises the largest urban forest in the world (> 12,000 ha). The local flora and fauna are protected by three conservation units and the Estação Biológica Fiocruz Mata Atlântica (EFMA), which comprises 462 hectares on the east side of the remnant. The local biodiversity is still little known compared to other Atlantic Forest remnants from the Rio de Janeiro State. Here, we provide results of a survey of medium- and large-sized terrestrial mammals from the EFMA. In addition, we analysed the distribution of this fauna along three habitat types defined as Peridomicile, Transitional Forest and Forest Core. Sampling was performed from 2017 to 2020 and comprised a camera-trap survey, interviews with residents and local workers and occasional records. Results include occurrence records for 16 autochthonous and one allochthonous (Callithrix sp.) wild mammals, which are distributed into 14 families and seven orders, in addition to the presence of free-ranging domestic dogs and cats. Four species are in some category of threat of extinction at national or global levels. Amongst them, Leontopithecusrosalia (first record for the Rio de Janeiro City in more than a century) and Leopardusguttulus are classified as Vulnerable by IUCN. Most wild native species were registered in the three habitat types, but with differences in the frequency of records. Our results indicate that the presence of domestic dogs and cats influenced the species composition in each area, with Nasuanasua, Dasyproctaleporina and Didelphisaurita less frequent in places where domestic dogs and cats are more frequent. This is the first systematic effort to understand the occurrence and distribution of mid- and large-sized mammals in the Pedra Branca Forest.

Bat species diversity from Reserva Ecológica de Guapiaçu, Rio de Janeiro, Brazil: a compilation of two decades of sampling

Reserva Ecológica de Guapiaçu (REGUA) is a private, protected area inside a remnant of the Atlantic Forest with high biodiversity. Although the bats of the reserve have been sampled for more than two decades, few studies have been published about them. Based on a compilation of data from several surveys, we present an updated list of the bat species there and compare it with surveys from other locations in the Atlantic Forest. From August 1998 to September 2021, at least 194 sampling nights were carried out at REGUA by different research groups from various institutions, totaling 448,092 m2.h of sampling. A total of 4,069 individuals were captured, belonging to 47 species and six families. Additionally, our results indicate that it is possible that some species that occur at REGUA have not been recorded yet. REGUA has the greatest number of bat species known for the Atlantic Forest. This most likely results from the fact that the reserve includes large areas of mature, continuous forest connected with other protected areas in the State of Rio de Janeiro. Another factor contributing to the high diversity of bat species at REGUA is that the area has been intensely sampled for many years. Given that the bat assemblage there appears to be a good proxy to ascertain the ecological patterns of biodiversity in well-preserved forests, we consider REGUA to be an important area for long-term ecological research. The basic knowledge about the ecological interactions of bats with different food resources and zoonotic microorganisms offers a unique opportunity to carry out research in several areas of knowledge, making it possible to address questions about bat assemblage structure, bat-parasite ecology, competition, niche partitioning, and other related studies.

Redescription of Myotis atacamensis (Chiroptera: Vespertilionidae) with neotype designation

Myotis atacamensis (Lataste, 1892) was described based on three syntypes from San Pedro de Atacama, Chile. The type series is lost. The original description was based on few external and cranial characters, and the diagnosis became obsolete and useless considering the current diversity of South American Myotis. Based on 12 specimens of M. atacamensis from southern Peru and northern Chile, we provide a morphological comparison with its South American congeners, designate a neotype, and provide a new diagnosis.(AU)

Trypanosoma spp. Neobats: Insights about those poorly known trypanosomatids.

Bats are infected with several trypanosomatid species; however, assessing the diversity of this interaction remains challenging since there are species apparently unable to grow in conventional culture media. Accordingly, the ecology and biology of the Molecular Operational Taxonomic Units (MOTUs) Trypanosoma spp. Neobats are unknown. Therefore, we performed the molecular characterization targeting the 18S small subunit rDNA from the blood clot of 280 bats of three Brazilian regions (Paraíba, Rio de Janeiro and Acre states), bypassing the selective pressure of hemoculture. From 68 (24%) positive blood clot samples, we obtained 49 satisfactory sequences. Of these successfully sequenced results, T. spp. Neobats (1, 3 and 4) represented 67%, with the most abundant T. sp. Neobat 4 (53%). Our results show: (1) high abundance and wide geographic range of T. sp. Neobat 4, restricted to Carollia bats; (2) high infection rate of T. sp. Neobat 4 in Carollia perspicillata populations (mean 26%); (3) infection with the monoxenous Crithidia mellificae; and (4) a new MOTU (T. sp. Neobat 5) in Artibeus cinereus, positioning in the Trypanosoma wauwau clade. These data corroborate the importance of bats as hosts of many Trypanosoma species and C. mellificae. They also show that the diversity of the T. wauwau clade is underestimated and warn about the high magnitude of trypanosomes we overpass with the hemoculture. Our findings combined with previous data show that T. spp. Neobats include host-specific and host-generalist species, probably playing different ecological roles: T. sp. Neobat 1 shows broad host range; T. spp. Neobat 3 and 4 are restricted to Artibeus and Carollia, respectively. Finally, T. Neobat 4 seems to be a well-succeeded parasite, especially within C. perspicillata metapopulations across a wide geographical distribution. This work is a step forward to understand the biology and life history of T. spp. Neobats.

Molecular eco-epidemiology of Paracoccidioides brasiliensis in road-killed mammals reveals Cerdocyon thous and Cuniculus paca as new hosts harboring this fungal pathogen.

Wild animals infected with Paracoccidioides brasiliensis represent important indicators of this fungal agent presence in the environment. The detection of this pathogen in road-killed wild animals has shown to be a key strategy for eco-epidemiological surveillance of paracoccidioidomycosis (PCM), helping to map hot spots for human infection. Molecular detection of P. brasiliensis in wild animals from PCM outbreak areas has not been performed so far. The authors investigated the presence of P. brasiliensis through nested-PCR in tissue samples obtained from road-killed animals collected nearby a human PCM outbreak spot, Rio de Janeiro state, Brazil and border areas. Eighteen species of mammals were analyzed: Dasypus novemcinctus (nine-banded armadillo, n = 6), Cerdocyon thous (crab-eating fox, n = 4), Coendou spinosus (hairy dwarf porcupine, n = 2), Lontra longicaudis (Neotropical river otter, n = 1), Procyon cancrivorus (crab-eating raccoon, n = 1), Galactis cuja (lesser grison, n = 1), Tamandua tetradactyla (collared anteater, n = 1), Cuniculus paca (paca, n = 1), and Bradypus variegatus (brown-throated three-toed sloth, n = 1). Specific P. brasiliensis sequences were detected in the liver, spleen, and lymph node samples from 4/6 (66.7%) D. novemcinctus, reinforcing the importance of these animals on Paracoccidioides ecology. Moreover, lymph nodes samples from two C. thous, as well as lung samples from the C. paca were also positive. A literature review of Paracoccidioides spp. in vertebrates in Brazil indicates C. thous and C. paca as new hosts for the fungal pathogen P. brasiliensis.

Crithidia mellificae infection in different mammalian species in Brazil.

Crithidia mellificae, a monoxenous trypanosomatid considered restricted to insects, was recently reported to infect a bat. Herein, C. mellificae has been demonstrated to have a wider range of vertebrate hosts and distribution in Brazilian biomes than once thought. Parasites isolated from haemocultures were characterized using V7V8 SSU rDNA and glyceraldehyde 3-phosphate dehydrogenase genes. Coatis (Nasua nasua) in the Cerrado; marmosets (Callithrix sp.) and bats (Carollia perspicillata, Myotis lavali, M. izecksohni, Artibeus lituratus) in the Atlantic Forest; crab-eating foxes (Cerdocyon thous) and ocelot (Leopardus pardalis) in the Pantanal biomes were infected by trypanosomatids that displayed choanomastigote forms in haemoculture in Giemsa-stained slide smears. Molecular characterization and phylogenetic inference confirmed the infection of C. mellificae in these animals. Moreover, slight differences in C. mellificae sequences were observed. Crithidia mellificae growth curves were counted at 27°C, 36°C and 37°C, and the morphotypes were able to grow and survive for up to 16 days. Serological titers for C. mellificae were observed in nonhuman primates, demonstrating that this parasite is able to induce a humoral immune response in an infected mammal. These results showed that host specificity in trypanosomatids is complex and far from understood.

The taxonomic status of Myotis nesopolus larensis (Chiroptera, Vespertilionidae) and new insights on the diversity of Caribbean Myotis.

Myotis nesopolus currently comprises two subspecies. The nominate subspecies (M. n. nesopolus) occurs on the Caribbean islands of Curaçao and Bonaire, Netherlands Antilles, whereas M. n. larensis is known from mainland South America in northeastern Colombia and northwestern Venezuela. Our Maximum Likelihood phylogenetic analyses of cytochrome-b gene sequences recovered M. nesopolus as a paraphyletic group, with M. n. nesopolus and M. n. larensis as non-sister lineages. The haplotype network indicates that these two subspecies do not share any haplotypes and are in different evolutionary trajectories. Additionally, these two subspecies can be distinguished on the basis of qualitative and quantitative morphological traits. This pattern supports the recognition of M. nesopolus and M. larensis as full species. Our results also reveal that the assemblage of Caribbean Myotis do not form a monophyletic group. Caribbean species are phylogenetically close to mainland species from northern South America and Central America, suggesting that colonization of Caribbean islands happened multiple times.

ResumoAtualmente Myotis nesopolus compreende duas subespécies: M. n. nesopolus ocorre nas ilhas caribenhas de Curaçao e Bonaire, Antilhas Holandesas, enquanto M. n. larensis é conhecido para o continente da América do Sul, no nordeste da Colômbia e noroeste da Venezuela. Nossa inferência filogenética por Máxima Verossimilhança recuperou M. nesopolus como parafilética, com M. n. nesopolus e M. n. larensis sendo linhagens não-irmãs. Além disso, essas duas subespécies não compartilham nenhum haplótipo. Adicionalmente, as subespécies podem ser diferenciadas a partir de caracteres morfológicos e morfométricos. Esse achado suporta o reconhecimento de M. nesopolus e M. larensis como espécies distintas. Nossos resultados revelam que os Myotis do Caribe não formam um grupo monofilético. Espécies caribenhas são filogeneticamente próximas de espécies continentais das Américas Central e do Sul, sugerindo que a colonização das ilhas do Caribe aconteceu por múltiplos eventos de dispersão.

Bats from the Pedra Branca Forest, Rio de Janeiro, Brazil.

The Pedra Branca Forest is located in a highly-urbanised region of the central portion of Rio de Janeiro City, comprises the largest urban forest on the continent and is isolated from other Atlantic Forest remnants. The local flora and fauna are protected by three conservation units (Pedra Branca State Park, Prainha Municipal Natural Park and Guaratiba State Biological Reserve) and one biological station (Fiocruz Atlantic Forest Biological Station-EFMA). Here, we provide an updated list of the bat fauna for the remnant. The results are based on samplings at EFMA and literature data from Pedra Branca State Park and Prainha Natural Park. The three sampling sites combined resulted in 31 species, 23 genera and four families. Phyllostomidae was the richest family with 24 species, followed by Vespertilionidae with five species (3%) and Molossidae and Noctilionidae with one species. The local bat fauna was predominantly composed of species with a broad geographic distribution.

Investigation of Bartonella spp. in brazilian mammals with emphasis on rodents and bats from the Atlantic Forest.

The Bartonella species are zoonotic agents that infect mammals and are transmitted by arthropod vectors. Approximately 18 distinct genotypes cause diseases in humans, and may be spread by both domestic and wild animals. In Brazil, Bartonella genotypes have been identified in several species of wild mammals, and in the present study, we analyzed samples from non-human primates (marmosets), marsupials, rodents, and bats, and compared them with the genotypes described in mammals from Brazil, to examine the distribution of Bartonella genotypes in two impacted areas of Rio de Janeiro state, in southeastern Brazil. We used polymerase chain reaction (PCR) methods to detect the Bartonella DNA using partial sequences of the gltA, ftsZ, and groEL genes. We generated Bayesian inference and maximum likelihood trees to characterize the positive PCR samples and infer the phylogenetic relationships of the genotypes. A total of 276 animals were captured, including 110 bats, 91 rodents, 38 marsupials, and 37 marmosets. The DNA of Bartonella was amplified from tissue samples collected from 12 (4.34%) of the animals, including eight rodents - Akodon cursor (5/44) and Nectomys squamipes (3/27) - and four bats, Artibeus lituratus (3/58) and Carollia perspicillata (1/15). We identified Bartonella genotypes closely related to those described in previous studies, as well as new genotypes in both the rodent and the bat samples. Considering the high diversity of the Bartonella genotypes and hosts identified in the present study, further research is needed to better understand the relationships between the different Bartonella genotypes and their vectors and host species. The presence of Bartonella in the wild rodents and bats from the study area indicates that the local human populations may be at risk of infection by Bartonella due to the spillover of these strains from the wild environment to domestic and peri-domestic environments.

Fiocruz Biological Collections: strengthening Brazil's biodiversity knowledge and scientific applications opportunities.

Biological collections are central in understanding and preserving life on Earth. In Brazil, the most representative collections are kept by natural history museums, whose primary focus is in invertebrates, vertebrates and vascular plants. Only a few institutions keep repositories in different kingdoms. The Oswaldo Cruz Foundation (Fiocruz), established in 1900, is a strategic public health institution of the Ministry of Health of Brazil. As such, Fiocruz is responsible for a wide range of activities, from basic research to the development and production of vaccines, drugs, reagents and diagnostic kits. Its biological collections were soon established in the expeditions made by naturalists and physicians seeking integrated knowledge of the fauna, flora and tropical diseases. Since then, they have been part of the institutional policy. In a few decades, those collections were already in the forefront of basic and applied research on tropical parasitic and infectious diseases. Currently, they comprise thirty-three repositories representing part of the Brazilian diversity of bacteria, fungi, protozoa, helminths, arthropods, molluscs and plants of medical and environmental importance. Different methods of long-term preservation are applied for the conservation of this wide range of organisms represented by about 6 million specimens. Herein, we describe this range of collections and discuss their complementary role as repositories of groups not represented in other biological collections in Brazil. These valuable biological materials have been used in public health and medical research, as well as for technological development and innovation in Brazil. Parallel to this specific usage, Fiocruz biological collections have played and continue to play a unique and important role in understanding and conserving part of Brazil's biodiversity that is currently under-represented in other biological and natural history collections in Brazil and South America.

The bat fauna (Mammalia: Chiroptera) of Carlos Botelho State Park, Atlantic Forest of Southeastern Brazil, including new distribution records for the state of São Paulo

Carlos Botelho State Park (PECB) is a large remnant of Atlantic Forest in Southeastern Brazil, with more than 37,000 ha. As its bat fauna is still unknown, we performed the first bat survey on PECB, to provide data on the distribution, natural history and taxonomy of the species. Fieldwork was conducted monthly, from October 2016 to September 2017. Captures were made using ground-level mist-nets (39600 m2.h), canopy mist-nets (2017.5 m2.h) and searches for roosts (42 hours).We captured 412 bats from 34 species of Phyllostomidae, Molossidae and Vespertilionidae. A total of 11 species were captured only in ground-level mist-nets, five in canopy mist-nets, and seven in roosts. Dermanura cinerea Gervais, 1856, Eptesicus taddeii Miranda, Bernardi & Passos, 2006, Glyphonycteris sylvestris Thomas, 1896 and Lampronycteris brachyotis (Dobson, 1879) are rare on surveys conducted in the Atlantic Forest of São Paulo and were captured in canopy mist-nets. Micronycteris schmidtorum Sanborn, 1935 and Molossus currentium Thomas, 1901 constitute the first record for the state of São Paulo, and were captured in canopy mist-nets and roosts, respectively. The species richness registered for PECB surpasses other surveys conducted in Atlantic Forest localities that use only ground-level mist-nets. Our results reinforce the importance of employing mixed capture methods, such as elevated mist-nets and searches for roosts.