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.

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.

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.

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.

Isolation and characterization of trypanosomatids, including Crithidia mellificae, in bats from the Atlantic Forest of Rio de Janeiro, Brazil.

We studied infection by Trypanosomatidae in bats captured in two areas with different degradation levels in the Atlantic Forest of Rio de Janeiro state: Reserva Ecológica de Guapiaçu (REGUA) and Estação Fiocruz Mata Atlântica (EFMA). Furthermore, we evaluated whether the diversity of trypanosomatids changes according to bat diversity and the different levels of preservation in the region. The results showed no influence of the level of preservation on bat species richness (15 and 14 species, respectively), with similar chiropterofauna and higher abundance of two common fruit-eating bat species in the tropics: Carollia perspicillata and Artibeus lituratus. Of the 181 bat specimens analyzed by LIT/Schneider hemoculture, we detected 24 infected individuals (13%), including one positive Sturnira lilium individual that was also positive by fresh blood examination. Molecular characterization using nested PCR targeting the 18 SSU rRNA-encoding gene fragment showed similar trypanosomatid infection rates in bats from the two areas: 15% in REGUA and 11% in EFMA (p = 0.46). Trypanosoma dionisii was the most frequently detected parasite (54%), followed by T. cruzi DTUs TcI and TcIV and Trypanosoma sp., in Neotropical phyllostomid bats (RNMO63 and RNMO56); mixed infections by T. dionisii/T. cruzi TcIII and T. dionisii/T. cruzi TcI were also observed. The T. cruzi DTUs TcI and TcIV are the genotypes currently involved in cases of acute Chagas disease in Brazil, and T. dionisii was recently found in the heart tissue of an infected child. Surprisingly, we also describe for the first time Crithidia mellificae, a putative monoxenous parasite from insects, infecting a vertebrate host in the Americas. Bats from the Atlantic Forest of Rio de Janeiro state harbor a great diversity of trypanosomatids, maintaining trypanosomatid diversity in this sylvatic environment.

Coxiella and Bartonella spp. in bats (Chiroptera) captured in the Brazilian Atlantic Forest biome.

BACKGROUND: The role of bats as reservoirs of zoonotic agents, especially pathogenic bacteria such as Bartonella and Coxiella, has been discussed around the world. Recent studies have identified bats as potential hosts of species from the proteobacteria phylum. In Brazil, however, the role of bats in the natural cycle of these agents is poorly investigated and generally neglected. In order to analyze the participation of bats in the epidemiology of diseases caused by Bartonella, Coxiella, Rickettsia, Anaplasma and Ehrlichia, we conducted a descriptive epidemiological study in three biogeographic regions of the Brazilian Atlantic Forest. RESULTS: Tissues of 119 bats captured in preserved areas in the states of Rio de Janeiro, Bahia and Santa Catarina from 2014 to 2015 were submitted to molecular analysis using specific primers. Bartonella spp. was detected in 22 spleen samples (18.5%, 95% CI: 11.9-26.6), whose phylogenetic analysis revealed the generation of at least two independent clusters, suggesting that these may be new unique genotypes of Bartonella species. In addition, four samples (3.4%, 95% CI: 0.9-8.3) were positive for the htpAB gene of C. burnetii [spleen (2), liver (1) and heart (1)]. Rickettsia spp., Anaplasma and Ehrlichia were not identified. This is the first study reporting C. burnetii and Bartonella spp. infections in bats from the Atlantic Forest biome. CONCLUSIONS: These findings shed light on potential host range for these bacteria, which are characterized as important zoonotic pathogens.

Range extension of Diclidurus scutatus (Chiroptera, Emballonuridae) to southwestern Amazonia / Extensão da distribuição de Diclidurus scutatus (Chiroptera, Emballonuridae) para o sudoeste da Amazônia

ABSTRACT Diclidurus scutatus is an aerial insectivore bat endemic of South America and considered rare throughout its distribution range. We present the first record of this species in Rondônia State, northern Brazil, expanding its distribution more than 1000 km into southwestern Amazonia. Including this record, D. scutatus is known for 20 localities from eight countries (Brazil, Colombia, Ecuador, French Guiana, Guyana, Peru, Suriname, Venezuela), and two biomes-Amazonia and Atlantic Forest.

RESUMO Diclidurus scutatus é um morcego insetívoro aéreo, endêmico da América do Sul e considerado raro em toda sua área de distribuição. Apresentamos o primeiro registro dessa espécie para o estado de Rondônia, norte do Brasil, expandindo sua distribuição em mais de 1000 quilômetros para o sudoeste da Amazônia. Incluindo o presente registro, D. scutatus é conhecido para 20 localidades em oito países (Brasil, Colômbia, Equador, Guiana Francesa, Guiana, Peru, Suriname, Venezuela) e dois biomas-Amazônia e Mata Atlântica.

Enhancing sampling design in mist-net bat surveys by accounting for sample size optimization.

The advantages of mist-netting, the main technique used in Neotropical bat community studies to date, include logistical implementation, standardization and sampling representativeness. Nonetheless, study designs still have to deal with issues of detectability related to how different species behave and use the environment. Yet there is considerable sampling heterogeneity across available studies in the literature. Here, we approach the problem of sample size optimization. We evaluated the common sense hypothesis that the first six hours comprise the period of peak night activity for several species, thereby resulting in a representative sample for the whole night. To this end, we combined re-sampling techniques, species accumulation curves, threshold analysis, and community concordance of species compositional data, and applied them to datasets of three different Neotropical biomes (Amazonia, Atlantic Forest and Cerrado). We show that the strategy of restricting sampling to only six hours of the night frequently results in incomplete sampling representation of the entire bat community investigated. From a quantitative standpoint, results corroborated the existence of a major Sample Area effect in all datasets, although for the Amazonia dataset the six-hour strategy was significantly less species-rich after extrapolation, and for the Cerrado dataset it was more efficient. From the qualitative standpoint, however, results demonstrated that, for all three datasets, the identity of species that are effectively sampled will be inherently impacted by choices of sub-sampling schedule. We also propose an alternative six-hour sampling strategy (at the beginning and the end of a sample night) which performed better when resampling Amazonian and Atlantic Forest datasets on bat assemblages. Given the observed magnitude of our results, we propose that sample representativeness has to be carefully weighed against study objectives, and recommend that the trade-off between logistical constraints and additional sampling performance should be carefully evaluated.

Karyotype of three Lonchophylla species (Chiroptera, Phyllostomidae) from Southeastern Brazil.

Lonchophylla Thomas, 1903 is a Neotropical bat genus that comprises 12 species, with little cytogenetic information available. Here we present the description of the karyotype of three species collected in Southeastern Brazil. Lonchophylla bokermanni Sazima, Vizotto & Taddei, 1978, Lonchophylla dekeyseri Taddei, Vizotto & Sazima, 1983, and Lonchophylla peracchii Dias, Moratelli & Esberard, 2013 showed the same diploid number 2n = 28 and the same autosomal fundamental number FNa = 50, in both Lonchophylla bokermanni and Lonchophylla peracchii. We observed that the karyotypes were also cytogenetically similar when we compared the studied species with other species within the same genus. It is therefore not possible to differentiate the species using only karyotypes with conventional staining. However, this information increases the knowledge of the genus and can be one more important character for a better phylogenetic comprehension of this taxon.

First evidence of frugivory in Myotis (Chiroptera, Vespertilionidae, Myotinae).

BACKGROUND: Myotis occurs from tropical to temperate regions throughout the globe, and it is the largest bat genus with more than 100 species. Most species are insect-eaters, but a few also feed on other invertebrates and fishes; there is no confirmed evidence of a plant item in their diet. NEW INFORMATION: During fieldwork in the Brazilian Atlantic Forest, small seeds were retrieved from the feces of one adult female of the Black Myotis, Myotis nigricans-one of the most common Neotropical bats. In a germination experiment, 40% of those seeds grew into seedlings. Our findings are the first evidence of fruit consumption for any Myotis species. We reject a possible contamination because the cotton bag was never used before for bats. This study is the first evidence of frugivory in the genus Myotis.

Bat assemblages from three Atlantic Forest fragments in Rio de Janeiro state, Southeastern Brazil.

Bat species richness in Neotropical localities is generally higher than that of any other group of mammals, and surveys of local bat assemblages may provide useful data for conservation management plans. Although the bat fauna of the Rio de Janeiro state is currently one of the best known in Brazil, there are several localities not adequately surveyed yet, and most of them are in the mountainous regions and in the northern portion of the state. From January 2008 to November 2009, we conducted surveys of bats in three localities in the state of Rio de Janeiro (municipalities of Varre-Sai, Sumidouro, and Cantagalo), and our fieldwork constitutes the first assessment of the bat assemblages of these localities. Surveys were conducted using mist nets in four different habitat types in each locality (forest interior, forest edge, riparian forest, and open areas [pastures]). We captured a total of 148 individuals in 17 species, 14 genera and 3 families. Among them, 11 species were recorded in Sumidouro, seven in Cantagalo, and nine in Varre-Sai. Although species richness was low compared with previous surveys in other close localities, we recorded species that have been rarely sampled in Southeastern Brazil (e.g., Macrophyllummacrophyllum [Phyllostomidae]). The results reinforce the importance of sampling different habitats in short surveys to improve the number of species registered.

Deslocamento de Artibeus lituratus (Olfers, 1818) (Mammalia, Chiroptera) entre ilha e continente no Estado do Rio de Janeiro, Brasil / Movement of Artibeus lituratus (Olfers, 1818) (Mammalia, Chiroptera) between island and continent on State of Rio de Janeiro, Brazil

Em 18 de julho de 2007, um indivíduo de Artibeus lituratus marcado com coleira plástica numerada foi recapturado no Parque Natural Municipal da Serra do Mendanha, município do Rio de Janeiro, 96 dias após ter sido capturado e marcado na Ilha de Itacuruça, no sul do Estado do Rio de Janeiro. A distância linear entre os pontos de marcação e recaptura sugere um deslocamento de no mínimo 35 km, sendo 500 m sobre o mar.

One individual of Artibeus lituratus, marked with a colored and numbered plastic color necklace was recaptured on July 18th, 2007 at the Parque Natural Municipal da Serra do Mendanha, Rio de Janeiro, Brazil, 96 days after being captured and marked at Itacuruça Island, located on southern Rio de Janeiro State. The linear distance between the plots of marking and recapture suggests a minimum movement of 35 km, being 500 m oversea.