High richness of non-volant mammals in a seasonal forest fragment in southeastern Brazil

The seasonal forest formations of the Atlantic Forest are a threatened and poorly known habitat. We present here a list of the non-volant mammals occurring in a 515-ha forest fragment known as Santa Maria and located in the Brazilian state of São Paulo. Our surveys are based on live trap captures, camera traps, and active searches for footprints, as well as secondary data. We list 29 species of non-volant mammals in the fragment, recorded between 1996 and 2021. One species found in the fragment, Leontopithecus chrysopygus, is globally endangered. Two species are classified as “vulnerable” in the global red list: Myrmecophaga tridactyla, and Tapirus terresris. One species, Panthera onca, is classified as “critically endangered” in São Paulo state. The red howler, Alouatta guariba, was not recorded after 1999 and has probably been extirpated in the fragment. We show that the number of non-volant mammal species in Santa Maria fragment is high, in relation to its size. The fragment is also in a strategic position, between the Morro do Diabo state park and the Black Lion Tamarin Ecological station, the two largest protected areas in the region. Considering its high mammal richness and its possible role as stepping stone for the local fauna, we recommend that the fragment become a protected area.


INTRODUCTION
The Atlantic Forest, a South American forest ranging from northeastern Brazil to northern Argentina and eastern Paraguay, is known for its elevated biodiversity and endemism (Mittermeier et al., 2011). It is also a highly threatened ecosystem, as only 28% of its original forest cover remains (Rezende et al., 2018;Ribeiro et al., 2009). If we consider the two main vegetal formations in the Atlantic Forest, the Dense Ombrophilous Forest and the Seasonal Semideciduous Forest, the latter has been much more devastated than the former, and in the state of São Paulo, only 6.4 % of its original cover remains (São Paulo, 2014).
The westernmost portion of São Paulo state, known as Pontal do Paranapanema or just Pontal, shelters the largest remnants of the Seasonal Semideciduous Forest of the state. Nearly 40,000 ha of this type of forest are inside protected areas such as the Morro do Diabo State Park (MDSP) with approximately 33,800 ha and the Black Lion Tamarin Ecological Station with approximately 6,700 ha, distributed in four sep-arated fragments (Faria & Pires, 2006;Valladares-Pádua, 2007). Besides these two large protected areas, there are in Pontal two private-owned forest remnants that harbor important portions of this endangered ecosystem: the Santa Mônica and Santa Maria fragments, each with an area of approximately 500 ha (Ditt et al., 1999).
Previous studies on the mammal fauna of the Pontal region have focused on the Morro do Diabo State Park and the Black Lion Tamarin Ecological Station, while the smaller fragments have received comparatively less attention (e.g., Faria & Pires, 2006;Valladares-Pádua, 2007). While some publications have provided a partial list of the mammals in some of the medium-sized fragments (e.g., Cullen et al., 2000), most of these data are in unpublished reports and dissertations (e.g., Ditt et al., 1999;Rocha, 2004;Tófoli, 2006).
The aim of this paper is to report the non-volant mammal species recorded in one of these areas, the Santa Maria fragment, and discuss its relevance for the conservation of the mammal fauna of the region and its associated ecosystem.

Study area
The Santa Maria forest fragment (hereafter SMA) is located in the municipality of Presidente Epitácio, in the westernmost region of São Paulo state, southeastern Brazil (22°14′S, 52°18′W, 289-344 m a.s.l.) (Fig. 1). The region is known as Pontal do Paranapanema, because in this area the Paranapanema river falls into the larger Paraná River. With an area of 515 ha, SMA is the sixth largest fragment of the Pontal do Paranapanema, and the largest one that is not located inside a protected area.
There are also several plant species in SMA that are exploited as food source by the non-volant mammals of the area. The jerivá palm, Syagrus romanzoffiana, a species known to produce a large amount of fruit constantly during the year (Begnini et al., 2013;Keuroghlian & Eaton, 2008), is a conspicuous palm in the fragment (Fig. 2). Other fruit trees include the araçá (Psidium sp.) and figs (Ficus spp.). Fruits of ground bromeliads, such as caraguatá (Bromelia sp.) and the roots of Smilax spp. are also consumed by terrestrial mammals in the area.
The SW border of the trapezoidal SMA is dominated by the exotic grass Brachiaria sp.; in the SE and NE, sugarcane plantation is the predominant land use (Figs. 1, 2). The NE border is limited by the country road SPV-035, a paved highway that connects Presidente Epitácio to the district of Planalto do Sul (Teodoro Sampaio municipality). A small creek, Ribeirão da Água Sumida, passes through the SE and SW borders of the fragment and falls into the large Rio Paraná (Figs. 1, 2).

Data collection
Our records of non-volant mammals are based on voucher specimens (small mammals traps), camera trap data, track records, direct sightings, and secondary data.
Small mammal traps were set up from January 2002 to December 2003. Sample was carried out in three trails, each 300 m long and parallel to each other (Rocha, 2004). Each of the trails had 12 sampling points. In each point, two live traps, one Tomahawk (44 × 20.5 × 21.5 cm) and one Sherman (23 × 9 × 8 cm) were set. Sampling effort totaled 1,924 trap nights and capture rate was 5.25%. To confirm the taxonomic identity of the small mammals, vouchers were collected and are deposited in the mammal collection of the Escola Superior de Agricultura Luiz de Queiroz (LMUSP), Universidade de São Paulo, Piracicaba, São Paulo.
Three Bushnell Trophy Cam camera traps were set from 7 th December 2014 to 3 rd February 2015, totaling 174 camera-days of sampling effort. Their placement was aimed at maximizing the sampled area (Table 1). The camera traps were not baited and were set on video mode (20 sec). Between 14 th February and 25 th February 2019, two Bushnell Trophy Cam camera traps were installed near the border of the fragment (Table 1), totaling 18 camera-days. We also installed seven camera traps in the canopy, which were set between 3 to 5 m high ( Table 1). The camera traps set above ground were originally part of a management action to monitor artificial shelters carried out by the Black Lion Tamarin Conservation Program/ IPÊ. The first canopy camera traps were installed on 5 November 2020 and, as of 28 August 2021, are still in the fragment. Licenses to capture and handle the animals Table 1. Coordinates and relative height to which the camera traps were placed in the Santa Maria fragment, Presidente Epitácio, Brazil. Mammal tracks on dirt paths surrounding the fragment were recorded ad libitum during incursions in the fragment for monitoring black lion tamarins from 2014 to 2015. Several species of mammals were opportunistically recorded while we were walking along a main trail of approximately 2,400 m long as part of our fieldwork with the lion tamarins.
Besides primary data, we used secondary data from IPÊ --Instituto de Pesquisas Ecológicas database (Ditt et al., 1999) and from the ATLANTIC-CAMTRAPS Datapaper (Lima et al., 2017). Searches in Google Scholar using "mammals" AND "Paranapanema" and "mamíferos" AND "Paranapanama" did not yield records for the Santa Maria fragment. We identified the photos and videos of the camera traps following Emmons & Feer (1998). The tracks were identified according to Becker & Dalponte (2013). The taxonomic arrangement follows the latest checklist of Brazilian mammals (Abreu-Jr. et al., 2021). Due to taxonomic uncertainties regarding the species of cottontails in western São Paulo, where both Sylvilagus minensis and S. paraguensis could occur, we refer to the taxon in SMA as Sylvilagus sp. (Silva et al., 2019). We consider the red brocket in the area to be Mazama rufa instead of M. americana, following Peres et al. (2021).

Data analysis
To verify which Atlantic Forest fragments, including protected and non-protected areas, from the same region were similar to SMA, we carried out a hierarchical cluster analysis using Euclidean distance. The community of medium and large mammals of SMA was compared with the ones from other 18 Atlantic Forest remnants (Fig. 3). Among these localities used in the comparison, we chose 11 Atlantic Forest fragments from the same broad region in western São Paulo (Ditt et al., 1999;Faria & Pires, 2006, 2010Lima et al., 2017). We also compared with four other localities of seasonal Atlantic Forest: the Iguaçu National Park (Silva, 2014), Amadeu Botelho Private Nature Reserve (Reale et al., 2014), Cosmópolis Forest (Magioli et al., 2014), and Morro Grande Forest Reserve (Negrão & Valladares-Pádua, 2006). The mammal community was also compared with three humid Atlantic Forest areas, the Carlos Botelho State Park (Brocardo et al., 2012), Intervales State Park (de Vivo & Gregorin, 2001), and Serra da Bocaina National Park (Delciellos et al., 2012). The localities to be compared were chosen to represent both the lowland seasonal and the humid montane formations of the Atlantic Forest. We selected montane localities in the Serra do Mar forest continuum of São Paulo and Rio de Janeiro and seasonal forest localities of São Paulo and Paraná states. Bearing in mind that differences in sampling methods and effort can insert biases the cluster analysis, we chose studies that used similar methodologies, i.e., active search for tracks, camera traps, and transects. For this analysis, we excluded small rodents (Cricetidae and Echimyidae) and marsupials (every genus except Didelphis).
To assess the richness of non-volant mammals in SMA relative to its area and to the neighboring forest fragments, we carried out a linear regression of species richness according to the fragment area for 14 fragments in western São Paulo state. For this, we included all non-volant mammals documented by us plus the species recorded by Ditt et al. (1999). All analyses were made in the R software (R Core Team, 2020).

RESULTS
A total of 29 species of non-volant mammals belonging to nine orders and 17 families were sampled (Table 2). We recorded nine species that are threatened in the state of São Paulo (SMA, 2018) and seven that are under some threat category in the Brazilian red list (MMA, 2014) ( Table 2). A total of three species are globally threatened (IUCN, 2021) ( Table 2). One species, the red howler monkey (Alouatta guariba) was last recorded in 1996-1998 surveys and has possibly been extirpated from the fragment. The jaguar (Panthera onca) is classified as "Critically Endangered" in São Paulo state (SMA, 2018) and the black lion tamarin (Leontopithecus chrysopygus) is classified as "Endangered" at local (SMA, 2018), national (MMA, 2014) and global levels (Rezende et al., 2020).
Hierarchical cluster analysis indicates that the SMA is more similar to the Ponte Branca (1,300 ha) and Tucano (2,115 ha) fragments, both part of the Black Lion Tamarin Ecological Station and 2.5 and 4-fold larger than SMA, respectively (Fig. 6). Every fragment of western São Paulo, except for Aguapeí State Park, grouped together (Fig. 6). The SMA contains more species of mammals than any fragment of similar size in western São Paulo, i.e., the Água do Peão, Santa Mônica, Seis R, and Lua Nova fragments (Fig. 7).

DISCUSSION
With 29 species recorded, the Santa Maria forest fragment harbors an unexpectedly high richness of non-volant mammals, despite its relatively small size (515 ha;   (Garbino et al., 2016). The high species richness of SMA may be explained by its proximity to the large Morro do Diabo State Park and by its position at the border between the Atlantic Forest and Cerrado ecosystems. Because our list is based on sampling efforts carried out along more than 20 years that used distinct methodologies, it was not possible to estimate the species richness in the fragment. However, when compared to what is known for the region (Ditt et al., 1999;Faria & Pires, 2006), we consider our list to be fairly complete. Still, some species, such as the Neotropical otter (Lontra longicaudis) and the naked-tailed armadillo (Cabassous tatouay) may be recorded in the remnant. We did not record the white-lipped peccary, Tayassu pecari. This species used to be relatively common in the Pontal do Paranapanema region (Cullen et al., 2000) but has be-come extremely rare, with the last confirmed sightings around 2008 and it has possibly been extirpated in the area (Fernando Lima, pers. com.). Field searches carried out by us (GSTG and GCR) between 2014 and 2015, in the three largest fragments of the region, i.e., the Morro do Diabo State Park, Tucano and Ponte Branca, also yielded no records of white-lipped peccaries.
Records of muroid rodents were also scarce, with only Oecomys cleberi recorded (Table 2). Although specimens of O. cleberi were collected in Santa Maria approximately 20 years ago, the taxonomic status of the species was ascertained only recently . Recent data, including specimens collected by us in other fragments of the region (Brandão et al., in review), suggest that O. cleberi is relatively common in Pontal. Nonetheless, we highlight that pitfall traps were not used by us or in previous studies (Rocha, 2004), which could hamper a proper evaluation of the local diversity of small rodents and marsupials.
Biogeographically, the mammal fauna in SMA has a mixture of Atlantic Forest and Cerrado taxa, which also can be explained by its geographic position, at the border between the two ecosystems. Typical Atlantic Forest taxa include Coendou spinosus, Alouatta guariba, and Leontopithecus chrysopygus, whereas Chrysocyon  (Brandão & Hingst-Zaher, 2021;Carmignotto et al., 2012). The ecotonal nature of the mammalian community in the fragment may explain its high species diversity, as has been shown in other studies carried out in areas with Atlantic Forest and Cerrado influences (Cáceres et al., 2007;Hannibal, 2014).     LC 1996-1998Ditt et al. (1999 Dasyprocta azarae Lichtenstein, 1823 x x x x DD DD LC 2019 Ditt et al. (1999), this study Hydrochoerus hydrochaeris (Linnaeus, 1766) x LC LC LC 2019 Ditt et al. (1999), this study

Erethizonthidae
Coendou spinosus (Cuvier, 1823) x LC LC 1996-1998Ditt et al. (1999 Lagomorpha Leporidae Sylvilagus sp.  Pap. Avulsos Zool., 2022;v.62: e202262017 8/11 Due to its strategic position, between two fragments of the Black Lion Tamarin Ecological Station with areas of over 1,000 ha, i.e., Água Sumida (1,199 ha) and the homonymous Santa Maria (2,057 ha), and distancing only 12 km of the large remnant of seasonal Atlantic Forest of the Morro do Diabo State Park, the SMA is an important stepping-stone for the local fauna. Also, considering the presence of endangered species, we recommend that the area integrates landscape connectivity plans along with the other fragments. The restoration of forest corridors between these forest remnants, which are less than 4 km apart (in a straight line) from each other, has the potential to establish an area of over 45,000 ha of continuous forest in the region. This can be very significant from the perspective of population viability of the threatened species that live in this fragment. We also recommend that the area be given the necessary attention by government authorities and be turned into a protected area, to ensure the protection and perpetuity of the fauna and flora of the fragment.

CONCLUSIONS
The high mammal richness and strategic location of the Santa Maria fragment makes it a priority area to be considered in the conservation of the fauna of Pontal do Paranapanema. We show that the fragment harbors endangered species and that some taxa have been extirpated from the fragment. Therefore, increasing its connectivity with neighboring areas is crucial. Future studies investigating mammalian diversity in the area, including bats and non-volant species, will also give a more complete picture of the mammal community in Santa Maria. Mazama sp.
x Ditt et al. (1999), this study * Possibly extirpated in the fragment.