Living and lost mammals of Rio de Janeiro’s largest biological reserve: an updated species list of Tinguá

The Tinguá Biological Reserve (TBR) is the largest protected area of this category in Rio de Janeiro state. Here, for the first time, we present the historical composition of terrestrial mammals’ assemblage of TBR region. An inventory was conducted using transect surveys, nonstandard transects, survey of museum specimens and informal reports. Considering all the data, eighty-five species were recorded, placing TBR as the second one in the number of mammals recorded in “Serra do Mar” ecoregion of Atlantic Forest and in the Rio de Janeiro state. Among the species with historical records are the jaguar (Panthera onca) and the golden-lion-tamarin (Leontopithecus rosalia) while the current presence of the maned wolf (Chrysocyon brachyurus) was recorded for the first time. Priority studies should focus on Chiroptera, Rodentia and Didelphimorphia orders, especially in the most remote areas of the reserve, and long-term surveys of endangered species. Besides hunting, fragmentation of its interior by roads, pipelines and transmission lines and exotic species, TBR is also threatened by the urban growth around it and the pressure to reduce its area and its protection category, demanding greater attention by the high levels of governance of protected areas in Brazil.

In Rio de Janeiro state, several mammal surveys found a high richness of small and medium species and the local extinction of large herbivores as Tapirus terrestris (Linnaeus, 1758), feeding guild specialists as Priodontes maximus (Kerr, 1792), Myrmecophaga tridactyla (Linnaeus, 1758) and top predators as Panthera onca (Linnaeus, 1758) (Araújo et al. 2008, Modesto et al. 2008a, 2008b, Delciellos et al. 2012, Carvalho et al. 2014, Aximoff et al. 2015. As a result of these impacts many populations show low densities and changes in behavior and activity patterns (Di Bitetti et al. 2008, Galetti et al. 2016. In most cases the Atlantic Forest mammal surveys are conducted with combined use of more than one sampling technique, such as camera traps surveys, interviews, linear transects and search by tracks and signs (Modesto et al. 2008a, 2008b, Silva & Passamani 2009, Espartosa et al. 2011, Delciellos et al. 2012, Carvalho et al. 2014. Most Atlantic Forest remnants are concentrated in the southeast region of Brazil, mainly in mountain hilltops of Serra do Mar. This mountain range cross the metropolitan zone of Rio de Janeiro in the "Baixada Fluminense" region (Custódio 2007), and this is the major refuge for wildlife in the surroundings of Rio de Janeiro city (MMA/IBAMA 2006). The Tinguá Biological Reserve is located in a megadiverse region of the Atlantic Forest. It has high plant diversity and richness (IPJB/UFRRJ 2002) and is equally important for the conservation of threatened and endemic species of birds, reptiles and amphibians (MMA/IBAMA 2006). For example, the Tinguá Biological Reserve is the only government-owned protected area in whole state of Rio de Janeiro where the Ornate Hawk-Eagle Spizaetus ornatus (Daudin, 1800) still occurs (Mendonça-Lima & Pacheco 2003). Although mammalian surveys performed at Tinguá neighboring protected areas showed high richness and the presence of endemic and endangered species (Cunha 2007, Olifiers et al. 2007, Carvalho et al. 2014, the composition of mammal species of Tinguá Biological Reserve remains poorly known.
The goal of this paper was to show what is known about the current and historical richness and composition of Tinguá's assemblage of terrestrial mammals. For this objective we performed an intensive field survey by linear transects. In a complementary way, we performed nonstandard sampling following trails, literature search, surveys in museum collection and gathered informal reports. These reports, about rare and shy species, were obtained with former poachers, local residents, rangers and other employees. Based on this information we recorded the degree of threat and discussed the importance of TBR to Atlantic Forest Mammals conservation and the necessity of long-term mammal surveys.

Study area
The Tinguá Biological Reserve (22º28'S and 22º40'S, 43º36'W and 43º13'W) (thereafter TBR) is the oldest protected area in Brazil; in 1833 emperor D. Pedro II created a Protection Forest to preserve the quality of the headwaters of three rivers, thus setting aside the area that later became TBR. Currently, TBR is administrated by "Chico Mendes Institute of Biodiversity Conservation" (ICMBio) -a government institute subordinate to the Ministry of the Environment. It is also the largest Biological Reserve in Rio de Janeiro state, with 248 km² spread over valleys and hills of Serra do Mar up to the altitude of 1600 m. The predominant vegetation is Dense Evergreen Atlantic Forest with a mix of mature and medium successional stages, and a canopy that reaches up to 25 meters. It has a high floristic diversity and the structure of the vegetation is considered one of the most well preserved in the whole Atlantic Forest biome (Iguatemy et al. 2017). The high terrain slope has prevented intensive logging. The east portion of TBR receives the rains that come from Atlantic Ocean. The mean annual temperature ranges between 20 and 26ºC and the annual precipitation is about 2000 mm ; there is no dry season (MMA/IBAMA 2006). The TBR is the core zone of a Biosphere Reserve of UNESCO, and close to other protected areas, like Araras Biological Reserve (38 km²), the Serra dos Órgãos National Park (200 km²) and Três Picos State Park (587 km²). This mosaic cover a total range of about 1.065 km² (MMA/IBAMA 2006).
Despite its high biological value -preliminary surveys has revealed that TBR harbors 350 species of birds and 52 species of amphibians (IBAMA 1996, MMA/IBAMA 2000) -TBR is subject to several anthropic impacts. Poaching and harvest of palm heart are frequent in the Reserve, due to its large perimeter and poor fiscalization (MMA/ IBAMA 2006). Livestook and horses from vicinity also invade the area occasionally and the reserve is crossed by electrical transmission lines and an oil pipeline (MMA /IBAMA 2006). During the last twelve years, the TBR buffer zone was subject of some deforestation due to logistic projects and other enterprises (Fundação S.O.S Mata Atlântica & INPA 2009.

Data collection
Between June 2005 and November 2006 a field survey was conducted using 10 linear transects spread throughout TBR forest. The length of transects ranged from 0.55 km to 2 km (average = 1.16 km; sd= 0.46 km) and the predominant vegetation was forests in late secondary successional stage. We started transect surveys from 30 minutes to one hour before sunrise and finished 3 or 4 hour after sunset. Transects were surveyed by one observer, rarely two, walking at 1 km/h, with brief stops each 50 or 100 meters. Besides that, opportunistic sightings and footprints were recorded from 2005 to 2009, either during the fieldwork carried out for this study or other field activities that together summed 1060 h. In the same period, we also recorded informal reports about the presence of medium or large mammals, cryptic or rare species with seventeen people that have a wide knowledge about the region, including former poachers, local residents that are in the region for over 20 years, TBR staff and researchers. During the study period, authorizations for research activities, which did not require catches for collections, were granted directly by the unit manager.
The mammal collection of the Museu Nacional at Rio de Janeiro was consulted in 2009 to record the species collected in the municipalities that harbor TBR (Nova Iguaçu, Duque de Caxias, Petrópolis and Miguel Pereira). A list of the specimens, summarizing all their information, was provided by the curators Dr. João A. Oliveira and Mr. Sérgio Maia Vaz. Only the specimens that were accurately identified in the list and do not have doubts about their identification were considered.
A literature search was conducted in specialized websites (e.g. Google Acadêmico, Scielo, Web of Science) aiming to compile faunistic surveys published up to 2016. To carry out the research we used key words "tinguá biological reserve", "mammals + reserva biológica do tinguá" or "reserva biológica do tinguá".
For each species we recorded the degree of threat using Bergallo et al. (2000) for the state of Rio de Janeiro, Chiarello et al. (2008) for the Brazil and the International Union for Conservation of Nature (IUCN 2015) for globally threatened species.

Results
A total of 433 kilometers walked on linear transects, 327 km diurnal and 106 km nocturnal, resulted in the visual records of 24 species of non-volant mammals. Occasional sightings and footprints recorded 22 species, adding ten species for those recorded through linear transects (Table 1). Three species were registered exclusively through reports, the neotropical otter Lontra longicaudis (Olfers, 1818), observed recently and the southern muriqui Brachyteles arachnoides (É. Geoffroy, 1806) and the jaguar Panthera onca (Linnaeus, 1758), recorded for the last time in the 1990s.
The total of mammals recorded in TBR corresponds to approximately 50% of the native terrestrial mammal species of the Rio de Janeiro state (Rocha et al. 2004), 27% of Atlantic Forest mammals and 13% of the endemic ones (Paglia et al. 2012). Twelve species (15%) recorded in TBR are in some threat categories at state level (Bergallo et al. 2000), seven species (9%) at national level (MMA 2014) and two species (2.6%) are in some threat categories of IUCN (IUCN 2017) (Table 1).

Discussion
This study reports the first comprehensive list of the mammals of the region of Tinguá Biological Reserve, providing the most complete "picture" of what we know about TBR's mammals. Despite the widespread defaunation scenery of Atlantic forest (Canale et al. 2012, Galetti et al. 2016, the TBR still harbor populations of large frugivores and apex predators like brocket deer, white lipped peccary and cougar, besides several mesopredators. TBR is one of the last shelters for large mammals in Rio de Janeiro and the last one in the northern portion of the Serra do Mar ecoregion. For example, TBR is the only refuge for white-lipped peccaries from the central region of the state up to the extreme north. No protected area north of TBR has currents sightings of the species in recent years (Araújo et al. 2008, Modesto et al. 2008a, 2008b, Carvalho et al. 2014). The red-brocket deer, by its turn, was recorded by one opportunistic sighting and three reports, one with a photo of an individual slaughtered inside the TBR (Figure 1). According to Duarte et al. (2016) Rio de Janeiro state is out of the geographic distribution of the species but our results together with the records in the Parque Nacional da Serra da Bocaina (Delciellos et al., 2012), Parque Nacional do Itatiaia (Aximoff et al. 2015) and Reserva Biológica Poço das Antas (Araújo et al., 2008) suggest that this interpretation should be revised.
The TBR is the second one in number of mammals already recorded in "Serra do Mar" ecoregion of Atlantic Forest and in Rio de Janeiro state, with 80 native species, exceeded only by Itatiaia National Park, and followed by Serra dos Órgãos National Park, with 106 and 75 species respectively (Table 2). However, these two protected areas are among the best sampled in Rio de Janeiro (e.g. Schirch 1932, Ávila-Pires & Gouveia 1977, Tribe 1987, Loretto & Rajão 2005, Cunha 2007, Macedo et al. 2007, Olifiers et al. 2007, so an increase in the sampling effort could reveal an even more important role for TBR. Small mammals, for example, are clearly underrepresented, as there is only one previous surveys carried out by Vaz (1984). The number of rodent species can be considered low when compared with other Serra do Mar study areas , Olifiers et al. 2007. Studies focused on these groups and the use of other methodologies not used in this study and able to detect inconspicuous species, like pit-falls and camera-traps (e.g. Srbek-Araujo & Chiarello 2005, Pardini & Umetsu 2006, could increase the number of species and are strongly recommended. For bats, the studies carried out by  and Lourenço et al. (2014) resulted in a number of species similar to other localities (Table 2). However, the absence of records of species present in other areas of Rio de Janeiro's Atlantic Forest (eg. Chrotopterus auritus, Trachops cirrohsus and Noctilio leporinus) also suggests that TBR is undersampled and that other species can be recorded with Table 1. Mammals recorded to the Tinguá Biological Reserve or around them, Rio de Janeiro state. The types of records were sighting in transection (T), opportunistic sighting (O), footprints (F), recorded in literature (L), museum collection (M) and report (R). For species registered only by reports the year in which the species was observed is provided in parentheses. Classification in categories of threatened species are according to Bergallo et al.(2000) at state level, Environment Ministry list (2014) at national level and IUCN (2015) at world level. Codes of threat degree are DD (deficient data), LC (least concern), PT (presumably threatened), NT (near threatened), VU (vulnerable), EN (endangered), CR (critically endangered), PEX (probably extinct) and EX (extinct). Species assigned with an asterisk are endemic of the Atlantic Forest. For species at the museum collection, the number of the considered exemplars are provided at the end of the  MN 5857, 5794, 5797, 5795, 5792, 5861, 5862, 5856, 5860, 5859, 5858, 5855, 5863, 5864, 5812, 5796, 5793, 5808, 5813, 5811, 42826, 50638, 51877, 51873, 51877, 51878, 28895, 28825, 28835, 28841, 5799, 5798, 5801, 5800, 28913, 50672 MN 4941, 4942, 4943, 4944, 4945, 4946, 4947, 4948, 4949, 4950, 4951, 4953, 4954, 8302, 21098, 34488  adequate sampling effort. In recent years, two new species of bats were assigned in TRB (Moratelli et al. 2011, Dias et al. 2013). Lonchophylla peracchii was described from specimens that had been first identified as L. bokermanni (Dias et al. 2013). Myotis izecksohni, previously identified as M. levis, had their types collected in TRB (Moratelli et al. 2011). This fact suggests that other new species can be hidden under cryptic taxa, reinforcing the importance of taxonomic studies.
Two species recorded by reports were no longer seen at TBR. The southern muriqui Brachyteles arachnoides (É. Geoffroy, 1806) was recorded in 1979 by an amateur ornithologist, Romildo de Mello, by a former poacher in the 1980s and by one of the authors in the 1990s, but its currently presence in TBR requires confirmation. The literature confirms the occurrence of the jaguar Panthera onca in Tinguá region in late 1960 decade (Silveira 1968) and we obtained two reports by former  poachers that described the killing of two individuals, an young male (whose carcass weighed approximately 30 kg) in the early 1980s and one adult in the early 1990s. The loss and fragmentation of the habitat and hunting pressure (and competition with human hunters by the same prey species) can have extinguished the jaguars in TBR. Another species previously recorded in the museum collection and not found nowadays was the golden-lion-tamarin Leontopithecus rosalia Linnaeus, 1766. The species was collected in Duque de Caxias municipality, locality of "Serra do Barro Branco" in 1939, near the area that would later become TBR. This record suggests that the lower slopes of TBR once housed a L. rosalia population, confirming the distribution of this species in the "Baixada Fluminense" (Coimbra-Filho 1969). This locality is in the same mountain range where a group of the species was recorded recently (Burity et al. 2007).
Some species which potential distribution includes the TBR and were never recorded in the area were the bush dog Speothos venaticus (Lund, 1842), a specialist carnivore, and large Insectivores and frugivores like Myrmecophaga tridactyla (Linnaeus, 1758) and Tapirus terrestris (Linnaeus, 1758) respectively. Although there are scarce records of lowland tapirs in Rio de Janeiro state (Schirch 1932, Ávila-Pires & Gouveia 1977, Spix & Martius 1981, the habitat of TBR is very similar of contiguous Serra dos Órgãos where the species was seen by early naturalists (Schirch 1932, Spix & Martius 1981. Therefore, it seems that these species are also locally extinct at TBR.
We also did not record medium and small species that have potential distribution for this portion of Serra do Mar including the nine-banded armadillo Dasypus septemcinctus, the marmoset Callithrix aurita, the titi monkey Callicebus nigrifrons, small rodents (Phylomys, Dellomys, Juliomys, Thaptomys) and some bats (Chrotopterus auritus, Trachops cirrohsus and Noctilio leporinus) among others species. These absences can be a methodological artifact, because few studies were long-term and some TRB areas, especially the most inaccessible stretches, were never sampled. However, the arrival of exotic invasive species can be an alternative explanation for the absence of some of them. Rattus rattus and Rattus norvergicus, for example, were reported in TRB for the first time in the 1940s. These rodents are effective competitors with small native rodents and can even prey upon some of them (e.g. Banks & Hughes 2012). Calithrix jacchus, by its turn, can also have negative impacts in other primate populations (e.g. Ruiz-Miranda et al. 2006). Besides these species, this study report for the first time the presence of Felis catus and Canis lupus familiaris within TBR. These animals, by their turn, have negative impacts on mammal population within protected areas (Galetti & Sazima 2006, Lessa et al. 2016. Inversely, the unexpected presence of the maned-wolf Chrysocyon brachyurus Illiger, 1815 was first documented in TBR. This species is typical from the cerrado but has been recorded in areas previously occupied by humid forests in the southeast coastal region (Queirolo et al. 2011). The process of savanization of the Atlantic Forest can be an important factor for the range expansion of this species (Sampaio et al. 2017).
The field sampling carried out in this study was useful to update the first TBR mammal list and show that species recorded by Vaz (1984) are still there. In comparison with other surveys in Atlantic Forest reserves (Cunha 2007, Norris et al. 2012, Rocha-Mendes et al. 2015, linear transects in TBR recorded more species of meso-predators. Besides the conservation status of the area, the record of many species of medium carnivores could be due to the use of local field workers, who are highly familiar with the forest and its local fauna, perceiving the signs of them easily, including the most inconspicuous ones.
In summary, our results show that TBR still maintains a high richness of mammals, and it is an important remnant to conserve medium and large-sized Atlantic Forest mammals. This patch was able to conserve populations of large game species like white lipped peccaries and can be a source of individuals to recolonize other areas, naturally or through reintroduction initiatives. It can also receive reintroduction initiatives provided that hunting pressure is controlled. Mammalogists and mammal ecologists have much work ahead, with low logistical investment due to its proximity to the large urban centers. Considering that hunting is still a significant presence in TBR interior, we recommend that population estimates of large mammals be produced and poaching effects investigated. The sampling of less studied groups such as Rodentia, Chiroptera, and Didelphimorphia would also be desirable. Most studies have been performed around a dirt road that cuts the TBR, while more remote areas remain without any sampling. The sampling of these areas and the use of the pitfalls, camera-traps and live-traps along all the altitudinal gradient, with appropriate sampling effort, could increase the number of species for the Reserve. All these efforts to know TBR species, however, will not be worth if the area continues to be neglected by the high levels of administration of protected areas in Brazil. Besides hunting, internal fragmentation and exotic species, TBR is threatened by the urban growth around it and the pressure to reduce its area and its protection level. If nothing is done to contain these threats, one of the best representatives of what was the mastofauna of the Atlantic Forest in the state of Rio de Janeiro may become just another example of an empty forest.