Reptiles of the Serra das Torres Natural Monument: using the Rapid Assessment method to fill a knowledge gap in the Atlantic Forest of southeastern Brazil using the to fill a gap the

: Data on the composition of local reptile assemblages in several Brazilian ecosystems can still be considered relatively restricted in scope in most cases. In this study, we conducted surveys in the Serra das Torres Natural Monument, located in the municipalities of Atílio Vivacqua, Muqui, and Mimoso do Sul, using the Rapid Assessments method (RAP) during 30 days in the rainy season of 2018. We sampled actively for approximately 1320 hours with a 6–10 person crew, supplemented by 720 hours of passive sampling (30 bucket-days) using pitfall traps with drift fences. We recorded 34 reptile species during our sampling method (2 amphisbaenid, 11 lizards, and 21 snakes) and an occasional encounter, after the end of sampling, that added a chelonian species to the list, Hydromedusa maximiliani , totaling 35 reptile species. The Dipsadidae was the family with the greatest snake species richness and, the Gymnophtalmidae had the greatest lizard species richness. The species richness recorded in the Serra das Torres Natural Monument (Ntotal = 35) represents ca. 27% of all reptile species found in the state of Espírito Santo (N = 130). The most abundant lizard species was Leposoma scincoides followed by Ecpleopus gaudichaudii and, the most abundant snake species was Bothrops jararaca being markedly higher than that recorded in similar studies. Twenty-seven percent of the reptile species recorded in our study are endemic to the Atlantic Forest and 30% (N = 10) have been recorded less than five times previously in the Brazilian state of Espírito Santo. Our study reinforces the need for the conservation of the Serra das Torres Natural Monument because of its importance as a reservoir of a considerable portion of the reptile biodiversity of Espírito Santo state, and of the Atlantic Forest biome as a whole.


Introduction
Data on the composition of local reptile assemblages in Brazilian ecosystems can still be considered relatively restricted in most cases. Although there is a reasonable number of studies on the reptile assemblages of some Brazilian forest remnants, most do not address the community as a whole, but rather, focus on a specific group, such as the snakes (Strüssmann & Sazima, 1993;Argôlo, 2004;Rocha et al. 2008;Pontes & Rocha 2008;Sawaya, 2008;Pontes et al. 2009;), lizards (e.g. Martins, 1991, or provide a compilation of the records available for an area, combining personal data with records obtained in previous studies (e.g. Vitt et al. 2008, Dias & Rocha 2014. However, comprehensive checklists of the reptile assemblage of a given ecosystem, based on a single, consolidated field inventory, are relatively rare, and most have focused on the rainforests of southeastern Brazil (Rocha, 1998;Almeida-Gomes et al., 2008;Rocha et al, 2018), although some important efforts are available for gallery forests in Central Brazil (e.g. Brandão & Araújo, 2001), and for the vast Lençóis Maranhenses region of northeastern Brazil (e.g. Miranda et al. 2012).
Understanding species composition, richness and abundance is a fundamental issue for countless conservation actions (Sutherland et al. 2013) and many ecological and taxonomic questions can be explored from this data set from a given area. Inventories of the reptile assemblages of the ombrophilous forest of Espírito Santo state, in southeastern Brazil, are restricted to a single introductory checklist of the reptiles of the Duas Bocas Biological Reserve (Tonini et al. 2010), in the municipality of Cariacica. Although there is an important set of specimens collected throughout the state, deposited in herpetological collections (e.g. Instituto Nacional da Mata Atlântica) efforts to discover the reptile fauna of the ombrophilous forest in Espírito Santo state remain as occasional collections, without short or long-term studies and either without information about the abundance of species.
The Serra das Torres Natural Monument, situated in southern Espírito Santo state (referred to here as the MONAST: Monumento Natural Serra das Torres) encompasses an array of upland environments characterized by high levels of biodiversity in some groups of fauna and flora (IPEMA 2010), but surprisingly there are no data available on its reptile assemblage. In this study, we inventoried the forests of MONAST through intensive sampling over a three-month period and we present the results here as the first comprehensive reptile checklist for the region of this pristine remnant of forest in the Brazilian state of Espírito Santo.

Study site
The MONAST is located in southern Espírito Santo state, and includes parts of the municipalities of Atílio Vivacqua, Mimoso do Sul, and Muqui, in southeastern Brazil (-21.0209, -41.2378) (Figure 1). The MONAST encompasses the largest complex of forest remnants in southern Espírito Santo, with approximately 10,450 hectares of Atlantic rainforest. This forest remnant includes mountains that reach 1100 m a.s.l. at their highest point (Oliveira et al. 2013) and the vegetation types are composed of semideciduous seasonal forest, submontane dense ombrophilous forest, and dense ombrophilous forest, all at varying levels of conservation (Magnago et al. 2008). The mean annual temperature is approximately 24.5ºC and mean rainfall is around 1290 mm (Oliveira et al. 2013).

Surveys of reptiles
We used the Rapid Assessment (RA) method to assess the species richness and abundance of reptiles in the MONAST during January, February, and March 2018. The RA method provides an efficient approach for the collection of reliable and replicable data in a short period of time (Patrick et al. 2014).
The RA method we employed in the present study consisted of intensive sampling over ten consecutive days at each of the three study sites located within the MONAST (a total of 30 field days) with a field team between six and 10 members, at 18 different locations. We conducted time-limited active searches (Crump & Scott Jr. 1994) between 0900 and 1200 hours for the daytime period, and between 1800 and 2200 hours for the crepuscular/nighttime period, with a total of approximately 1320 hours of sampling effort. We conducted the active searches in preserved forest fragments located as far as possible from areas of anthropogenic impact, including areas at altitudes ranging from ca. 600 m to 1100 m a.s.l. To guarantee the best possible estimate of local species richness, we also recorded the reptile species found in the habitats in the vicinity of the MONAST, including human habitations, pasture, and plantations. We did not conduct an active search more than once at the same geographic coordinate.
We also collected reptiles using four systems of pitfall traps with drift fences (Corn 1994) installed in different forest fragments, in particular the best-preserved habitats, and at different altitudes. Each system consisted of 40 buckets (of 20 L), which were arranged in a straight line at each study site.
We installed the traps three days prior to the sampling period at each of the three sites, with all buckets remaining active until the 10th consecutive day of sampling. We removed all the buckets and fences at the end of the sampling period at each site and then moved them to the next sampling area. The overall sampling effort of the pitfall traps was approximately 720 hours. At the end of the survey, we removed all the pitfall systems from the MONAST.
We identified the reptile species using specific references (e.g., Ávila-Pires 1995; Marques et al. 2001;Silva Jr et al. 2016) with confirmation by specialists, whenever necessary. The taxonomic identity of the species known to occur in Espírito Santo was confirmed through consultations at the herpetological collections of the National Atlantic Forest Institute (INMA: Instituto Nacional da Mata Atlântica), in Santa Teresa, Espírito Santo state, and the Museu Nacional (NMRJ) in Rio de Janeiro state, in order to minimize possible errors with regard to species occurrence. We considered these two collections to represent accurately the herpetofauna known to occur in the Brazilian state of Espírito Santo.
We constructed a species accumulation curve based on the cumulative number of species recorded during the RA (S) as a function of sampling effort (n). We estimated species richness by the Bootstrap method, which we considered to be the diversity index best suited to our data (Magurran 2004). We also classified species occurrence as "rare" or "common" based on the number of previous records from Espírito Santo, using the data available in online databases (Species Link 2018). Based on these previous records from Espírito Santo state, we classified species in four classes of occurrence: (i) 1-5 occurrences, (ii) 6-10 occurrences, (iii) 11-20 occurrences, and (iv) more than 20 occurrences. We considered species in the (i) 1-5 occurrences category to be "rare" in the state of Espírito Santo.
The collection of voucher specimens was authorized by Sisbio/ RAN Nº 57085-6 and the Espírito Santo for the Environment (IEMA) Nº 033-2017. The vouchers were deposited at the MNRJ, in Rio de Janeiro and INMA, in Espírito Santo state. We did not collect specimens of some easily-recognized species (e.g., Salvator merianae, and Boa constrictor) due to their large body size, which would require additional resources for specimen collection.

Results
We obtained 257 specimens of 35 reptile species representing 15 families distributed in two orders, Chelonia (N = 1) and Squamata (N = 34) during our inventory of the Serra das Torres Natural Monument (Table 1, Figure 2 and 3). The Squamata species recorded were represented by two amphisbaenids, 11 lizards, and 21 snakes.  The snake family with the largest number of species was the Dipsadidae (N = 13 species), while in the case of the lizards, the most diverse family was the Gymnophtalmidae (N = 4 species). The most abundant lizard species in our inventory were Leposoma scincoides (N = 54) and Ecpleopus gaudichaudii (N = 38), followed by Enyalius boulengeri (N = 27) (Figure 4). The rarest lizard species were Heterodactylus imbricatus and Ophiodes fragilis, both recorded only twice and Placosoma glabellum and the exotic, invasive gekko Hemidactylus mabouia, which were both recorded only once (Figure 4). The most abundant snake species was Bothrops jararaca (N = 47 specimens), while the other 21 snake species were all recorded less than five times during the study ( Figure 5). We recorded two species of Amphisbaenia, Amphisbaena alba (N = 1) and one Leposternon microcephalum. We also recorded one Hydromedusa maximiliani after the end of the RA sampling.
This chelonian was included in the MONAST species list and richness but was not included in the analyses. Additionally, we did not collect a voucher specimen of H. maximiliani due to the threatened status (vulnerable) of the species in Espírito Santo state (IPEMA 2007;Bérnils et al. no prelo). The species accumulation curve did not reach the asymptote, although the total species richness estimated for the MONAST based on the Bootstrap index was 36, only slightly above the actual species richness recorded in our study (34 species) ( Figure 6). The largest number of species was recorded by active searching (N = 24 species), whereas only eight species were collected in the pitfall traps (Table 1). Four species (three lizards and one snake) were recorded exclusively by the pitfall traps.
Eight of the 11 lizard species (72%) recorded in the MONAST are endemic to the Atlantic Forest biome and two of the snakes are also endemic to this biome (Table 1). Ten (30%) of the reptiles recorded in the present study had less than five previous records from the state of Espírito Santo, and were thus classified as "rare", and the same number (10 species, 30%) had been recorded more than 20 times, and were thus classified as "common" (Figure 7).

Discussion
The reptile species richness recorded in the MONAST (N total = 35) represents ca. 30% of all the squamates found in the Brazilian state of Espírito Santo (N = 117, see Costa & Bérnils 2018). This indicates that the MONAST protects a large proportion of the overall reptile biodiversity of Espírito Santo and, in turn, of the Atlantic Forest biome. The species richness of Serra das Torres is also similar to other remnants ombrophilous forest in southeastern Brazil where reptile surveys have been conducted. For example, Moura et al. (2012)      (one chelonian, one crocodilian, 10 lizards, and 24 snakes) in the Guapiaçu Ecological Reserve, in Rio de Janeiro. It is important to note that our inventory recorded 34 species (excluding H. maximiliani, found after the end of sampling), while the predicted species richness was only a little higher (36 species), despite the fact that the accumulative species curve did not reach the asymptote. These findings support the reclassification of the MONAST remnants as a more restrictive category of conservation unit, such as a state park, to better guarantee the protection of the reptile diversity of this important Atlantic Forest complex. Hemidactylus mabouia, a non-indigenous lizard, was recorded in a forest fragment next to a banana plantation at the borders of the MONAST. This gekko is native to Africa, and has invaded a number of different natural environments in Brazil , Telles et al. 2015, Oliveira et al. 2016, including many ombrophilous forest remnants Telles et al. 2015). The occurrence of H. mabouia in the MONAST is consistent with the dispersal of the species in other regions, where it is associated with open areas impacted by human activities Oliveira et al. 2016). The record of H. mabouia collected in the present study suggests that this species may begin to invade MONAST's natural environments.
The dominance of the family Dipsadidae in the present study is consistent with the pattern found in other areas of the Atlantic Forest (e.g. Hartmann et al. 2009, Freitas 2014. The Dipsadidae includes most tropical snakes, with more than 700 species (Vidal et al. 2010, Uetz et al. 2019, and the predominance of dipsadids recorded in the present study indicates a conservative snake community structure in Serra das Torres. Gymnophthalmidae was the most diverse lizard family (N = 4 species) in MONAST. This family as composed of small lizards, typically semifossorial, which forage in the leaf-litter (Dixo and Verdade 2006). In our samplings, gymnophthalmids were captured mainly in the pitfall traps, what is in accordance with the habitats of most species.
The most abundant lizards in our study were Leposoma scincoids and Ecpleopus gaudichaudii, whereas Heterodactylus imbricatus and Placosoma glabellum were the least abundant in the community. This result is comparable with other studies in Atlantic Forest remnants were the species of the Gymnophthalmidae family are major components of local leaf-litter reptile communities (e.g. Maia et al. 2011, Cruz et al. 2014. For example, E. gaudichaudii represented a large proportion of the leaf-litter reptile community in the Morro Grande Forest Reserve, in São Paulo state (54%), whereas Heterodactylus imbricatus was the least abundant lizard (3%) (Dixo and Verdade 2006). In the MONAST, 32% of the lizards recorded during the survey were Leposoma scincoides, and 23% were E. gaudichaudii while H. imbricatus represented only 1.2% and Placosoma glabellum only 0.6% of the lizard's abundance. These four lizard species are endemic to the Atlantic Forest of southern and southeastern Brazil (Tozetti et al. 2017) and are considered species dependent on forested areas (Dixo and Verdade 2006). Since these species depend on preserved areas, local changes in the environment, even on a small scale, can result in changes in the composition or even the local extinction of these species (Román-Cuesta and Martínez-Vivalta 2006).
The most abundant snake was Bothrops jararaca. This pitviper has a wide distribution in the Atlantic Forest, occupying well-preserved, impacted, and anthropogenic areas (Sazima 1988, 1992, Campbell and Lamar 1989. Although B. jararaca is a common snake in the communities in which it occurs, its abundance varies considerably among localities. In Serra do Mendanha, in Rio de Janeiro state, for example, B. jararaca is relatively rare, with only 10 records being obtained in a 62-month study (Pontes and Rocha 2009), whereas in the Juréia-Itatins Ecological Station, in São Paulo state, the species was relatively abundant, with 60 records being collected during a 15-month study (Marques and Sazima 2004). In the MONAST, Bothrops jararaca was more abundant (N = 47 records in 30 days of sampling) than all the other snake species combined (N total = 42). Although it is important to consider the differences in the methods adopted in the different studies, the abundance of B. jararaca in Serra das Torres indicates that this forest remnant is a potentially important site for the conservation of this pitviper species.
We also recorded nine reptile species (27% of the total) that are endemic to the Atlantic Forest in our study at MONAST (Table 1).
Overall, approximately 100 species are endemic to this biome (Tozetti et al. 2017). The Atlantic Forest is one of the world's most biodiverse biomes, although it is also one of the most threatened, reinforcing its status as a conservation hotspot (Myers et al. 2000, Ribeiro et al. 2009, Colombo and Joly 2010. Additionally, one chelonian (Hydromedusa maximiliani) and one snake (Tropidophis paucisquamis) are considered as vulnerable (VU) in the red list of threatened species in the state of Espírito Santo (Bérnils et al. no prelo). Hydromedusa maximiliani is endemic to the Atlantic Forest domain, where it inhabits streams at high altitudes (above 600 m a.s.l.) in the states of Bahia, Espírito Santo, Minas Gerais, São Paulo, and Rio de Janeiro (Argôlo & Freitas 2002). The main threat to the conservation of H. maximiliani is the alteration of the vegetation cover, which modifies the species' natural habitat and provokes shifts in the temperature beyond the optimum it requires to survive (Almeida et al. 2007). Hydromedusa maximiliani is also listed as Vulnerable in the IUCN Red List (IUCN 2019). Tropidophis paucisquimis is endemic to Atlantic Forest, occurring in the states of Espírito Santo, Rio de Janeiro, São Paulo, Paraná and Minas Gerais  and are mainly associated with the Serra do Mar and Serra da Mantiqueira mountain chains at an altitudinal range from 500 to 1261 m. (Curcio et al. 2012). The dwarf boa T. paucisquamis is rare in inventory studies and there are only four records of the species in Espírito Santo state (this study) and only 50 specimens from 26 localities are deposited in scientific collections of the entire Brazil . This species has among its antipredator mechanisms the cephalic autohemorrhage which has been described for species of the Tropidophis genus as a physiological product of increased blood pressure associated with a behavioral response caused by stress or fright (Smith et al. 1993). However, information about the biology and natural history of Tropidophis paucisquamis are poorly known. The presence of a high number of endemic species, habitat-dependent species and endangered species make MONAST an important refuge for the reptile fauna of the Atlantic forest.
In addition, 10 (30%) of the reptiles recorded in our study have less than five previous records in Espírito Santo (Figure 7). The rarity of these species in the state probably reflects in many cases the lack of studies in this region of southeastern Brazil. For example, Micrurus lemniscatus carvalhoi, is known to occur in western Argentina and eastern Paraguay, and is widespread in most Brazilian biomes, although only two previous records were available from Espírito Santo (Castro et al. 2017). This is typical of most of the rare species recorded in the present study and, considering the ongoing deforestation in Espírito Santo, we would strongly recommend that further studies of the state's herpetofauna be carried out.
The importance of the MONAST for the protection of the region's herpetofauna only began to be revealed in the past decade (e.g. Oliveira et al. 2009Oliveira et al. , 2012Oliveira et al. , 2013, and the new data presented here further reinforce the importance of this Atlantic Forest remnant for the conservation of the Atlantic Forest herpetofauna. The present study is the first inventory for southern Espírito Santo and it has been fundamental to the filling of a large knowledge gap in this portion of Atlantic Forest. Finally, we would recommend that the local environmental authorities focus on the need to maintain the integrity of the habitat structure of the entire remnant that harbors the Monumento Natural Serra das Torres.