Anurans from the Serra da Bocaina National Park and surrounding buffer area, southeastern Brazil

Here, we review the species of anuran amphibian from the Serra da Bocaina National Park and its buffer area, in the Atlantic Forest of southeastern Brazil, comparing the data from a recent survey with museum records. We surveyed adult and larval anurans in ponds, marshes, and streams discontinuously from May 2008 to January 2011. In total, 63 anuran species were previously known to occur at the Park and its surrounding buffer area. In our survey, we recorded 46 species, of which five represented new records, and 22 appeared only in the historical list. Seven topotypic populations were not found in the present study. We suggest that conservation strategies for anurans in the Serra da Bocaina should also consider the surrounding areas that are subjected to anthropogenic pressure, due to the high diversity recorded, high altitudinal variation in species distribution, and various vegetation formations.


Introduction
Brazil is the home of 968 amphibian species, of which 932 are anurans (AmphibiaWeb 2014). The highest amphibian richness in Brazil occurs along the costal Atlantic Forest, to which many species are endemic (Rossa-Feres et al. 2011;Haddad et al. 2013). This high richness is in part due to the environmental heterogeneity created by the various vegetation types, the altitudinal, and longitudinal and latitudinal variation along this biome (Haddad and Prado 2005;Araujo et al. 2009;Ribeiro et al. 2009), which limit the dispersal and favor the occurrence of species with small range sizes (Villalobos et al. 2013). Furthermore, the climatic refugia of the Pleistocene and neotectonism (Thomé et al. 2010) seem to play a key role in dictating the contemporary species richness of several taxa (Carnaval and Moritz 2008).
The Atlantic Forest is composed of two main vegetation types: Atlantic Rain Forest and Atlantic Semi-deciduous Forest (Morellato and Haddad 2000). The Serra da Bocaina National Park is a protected area in the Atlantic Forest in the Serra do Mar, at the border between the states of São Paulo and Rio de Janeiro, in Southeastern Brazil. The Park currently comprehends an area of 140,000 ha, from the sea level up to 2,088 m a.s.l. and is covered by the Atlantic Rain Forest, with natural and anthropogenic grasslands in high altitudes. Although many collections of amphibians have been carried out in the past (1925 to 1970), mainly by Adolpho and Bertha Lutz, and Werner C. A. Bokermann, no long-term ecological study encompassing an entire reproductive season had been conducted. These past research efforts resulted in 17 anuran species being described from the Park and its surrounding buffer area, which includes the Fazenda Bonito (Lutz 1925;Bokermann Abstract: Here, we review the species of anuran amphibian from the Serra da Bocaina National Park and its buffer area, in the Atlantic Forest of southeastern Brazil, comparing the data from a recent survey with museum records. We surveyed adult and larval anurans in ponds, marshes, and streams discontinuously from May 2008 to January 2011. In total, 63 anuran species were previously known to occur at the Park and its surrounding buffer area. In our survey, we recorded 46 species, of which five represented new records, and 22 appeared only in the historical list. Seven topotypic populations were not found in the present study. We suggest that conservation strategies for anurans in the Serra da Bocaina should also consider the surrounding areas that are subjected to anthropogenic pressure, due to the high diversity recorded, high altitudinal variation in species distribution, and various vegetation formations. Park, which includes areas of the cities of Arapeí, Areias, Bananal, Cunha, São José do Barreiro, Silveiras, Ubatuba in the state of São Paulo, and Angra dos Reis and Paraty in the state of Rio de Janeiro. We surveyed tadpoles and adult anurans in 14 breeding sites monthly (May 2008to June 2009, and November 2009to February 2010, including ten ponds, three marshes, and one stream. These water bodies ware sampled once during one week each month. We also surveyed other ten breeding sites (six streams and four ponds) occasionally. Tadpoles were sampled with hand dipnets along the entire margins of water bodies, with effort proportional to surface area (Skelly and Richardson 2010). To survey adult anurans, we employed three different and complementary methodologies: visual encounter surveys, survey at breeding sites (Scott and Woodward 1994), and also searched for direct-developing species on the forest floor and bromeliads, near water bodies or along trails. In every breeding site, we recorded calling males as well as adults that were not vocalizing. Further environmental data on sampling sites are available in Provete (2011). Pictures from the sampling points, along with their geographical coordinates, are available online at CalPhotos (bitly.com/ Bocaina). A Google Earth file with sampling points is available at diogoprovete.weebly.com/publications.
To assemble the historical list of species, we searched for museum records and in the literature. Literature search took into account the locality (e.g., Parque Nacional da Serra da Bocaina, Campo de Fruticultura, município de Bananal). Museum search included Museu de Zoologia da Unicamp (ZUEC-AMP; Campinas, Brazil) and Coleção Célio F. B. Haddad (CFBH; Rio Claro, Brazil) available online at www.splink.cria.org.br (captured on 10 January 2014), besides the amphibian collection of the Smithsonian National Museum of Natural History (USNM; Washington D.C., United States). Since this study was not designed to be a taxonomic revision, we did not checked the identity of each museum specimen, except for the specimens housed at the CFBH collection. We checked each record of the other museums to exclude any outlier or those species that are not know to occur in the Serra da Bocaina using the available geographic coordinates. Furthermore, uncertain identifications, such as "sp.", "affinis", or "group" were also excluded from our list. We looked for literature and catalogs mentioning material collected in the Serra da Bocaina and its buffer area (Lutz 1925;1929;Lutz and Lutz, 1938;1939;Lutz in Lutz 1950;Cochran 1955;Bokermann 1966;1967a,b;Heyer 1985;Cruz and Peixoto 1984;1985;Peixono 1989;Giaretta, Bokermann and Haddad, 1993;Pombal and Hadad 1999;Pombal 2001;Napoli and Caramaschi 2004;Almeida and Angulo 2006;Prado and Pombal 2010;Martins and Zaher 2012;Frost 2013, Caramaschi andCruz 2013; Appendix 2). In the case of the genus Fritziana, the historical list includes four species. However, those species in the genus occurring in southern Brazil seem to be a species complex, whose members are difficult to tease apart. Therefore, we took a conservative decision and list only one species. the nomenclature used here follows Frost (2013).
We run a Jackknife 2 to estimate the species richness based on the incidence of larvae and adults. We used this estimator because it can handle unequal sampling effort among habitats and is biased to species that occur in a few samples (Hortal et al., 2006). Data were analyzed using software EstimateS ver. 8.2 (Colwell, 2006).

Results
Sixty-three anuran species were previously known from the Serra da Bocaina National Park and its surrounding buffer zone (Table 1; Figures 2-3). In this study, we recorded 46 species (Jackknife 2 = 53 species), of which 41 also appear in museum records and five species represent new records from the Serra da Bocaina National Park. These records increased the list of the area to 68 species.
According to the IUCN Red list of threatened species (IUCN, 2008), 13 species (19%) are classified as Data Deficient (DD), whereas the remainig species were either Not Evaluated (NE) or not under any threatened status (LC). The São Paulo state red list (Garcia et al. 2009), includes 4 species as threatened: Crossodactylus dispar is endangered (EN), Paratelmatobius gaigae is vulnerable (VU), and Bokermannohyla ahenea and Physalaemus barrioi are near threatened (NT).

Discussion
The anuranfauna currently known in the Serra da Bocaina National Park and surrounding buffer areas represents approximately 13% of the species known to Atlantic Forest biome (Rossa-Feres et al. 2011;Haddad et al. 2013). Considering only the species richness along the Serra do Mar, the anuran diversity in the Park is similar to other localities with high altitudinal variation, such as the Boracéia Biological Station (65 species; Heyer et al. 1990) and Carlos Botelho State Park (65 species; Forlani et al. 2010). The high diversity along this mountain range is probably due to its high altitudinal variation, which favor the high species turnover along altitudinal gradients. Species turnover along altitudinal gradients in the Atlantic Forest has already been found in several groups (e.g., Almeida-Neto et al. 2006), including anurans (Giaretta et al. 1999. The greatest sampling efforts, both in the past and in the present study, was concentrated on the northern areas, above 1,000 m a.s.l. In this area, we record the highest species richness. These data reinforce the need to increase sampling efforts in the southern region of the Park and areas of lower altitude, which certainly will prove fruitful. Some topotypic populations and species known from museum records were not found in the current surveys. The majority of these species occur in the highlands, such as Bokermannohyla clepsydra, Megaelosia bocainensis, Crossodactylus dispar, Cycloramphus granulosus, Paratelmatobius gaigeae, Vitreorana eurygnatha, and Ischnocnema pusilla. Some species present direct development on the forest floor (Brachycephalus vertebralis and Holoaden suarezi, Haddad and Prado 2005), or are stream breaders (C. dispar and V. uranoscopa, Haddad and Prado 2005), which were habitats poorly sampled in our study. Since our sampling effort was concentrated in high altitude areas, widely distributed species, such as Hypsiboas faber, Leptodactylus labyrinthicus, and Leptodactylus mystacinus and also those known to occur in lower altitude areas (Serafim et al. 2008) were not recorded. The specific locality in which Bokermann (1972) found B. clepsydra, andGiaretta et al. (1993) found M. bocainensis, called Campo de Friticultura (also known as Fazenda Ponte Alta) was surveyed for 14 consecutive months. However, we find neither adults nor tadpoles of these species. Probably, the lack of current records of B. clepsydra might be due to environmental changes, occurred since this area passed to the Park administration in the late 1970s. After that, the forest expanded and the landscape changed dramatically, which might have contributed to the local decline of this population. The same seems to be the case for other endemic species, which were known to inhabit the Campo de Fruticultura, such as Physalaemus barrioi that now is found only in open areas in the buffer area of the Park (Provete et al. 2012).
The two survey techniques employed in this study demonstrated to be complementary. We recorded four species only by sampling tadpoles (Phasmahyla cochranae, Scinax ariadne, Scinax crospedospilus, and Thoropa taophora). While 20 species were only recorded by adult sampling, and 22 species were found as both larvae and adults. Both methods contribute to a more accurate sampling of the anuran richness if applied together, as already pointed out by a recent study (Silva 2010).
We recorded the majority of species in forested environments, including most of the endemic species in the study area (Table 1). However, the high species richness in grasslands, with some of the species restricted to this environment (e.g., P. barrioi), highlight the need for conserving this type of environment, which currently is under high anthropogenic pressure in the surrounding buffer area.
The conservation status of most species was not evaluated yet, mainly due to lack of detailed information about natural history (IUCN 2008;Garcia et al. 2009). Bokermannohyla ahenea and P. barrioi are both classified as Near Threatened and are also endemic to the Serra da Bocaina highlands. Two species listed as threatened in the state redlist (Garcia et al. 2009) are found in the Park: C. dispar (EN) and P. gaigeae (VU). Crossodactylus dispar is widely distributed, occuring in the states of Rio de Janeiro, São Paulo, and Minas Gerais. Paratelmatobius gaigeae is also endemic to the study area, occurring at the Fazenda do Bonito and Bananal Ecological Station (Zaher et al. 2005;Frost 2013). None of the former species were recorded by us, and more field work aiming specifically to collect those species seems to be necessary to evaluate their conservation status properly and extent of occurrence. In this study, we provided a list of anuran species, mainly for the northern portion of the Serra da Bocaina considering the current survey and museum data. Information about species richness and composition and current population trends are relevant to subsidize management plans and conservation strategies (Colombo et al. 2008). Furthermore, data about species distribution can have an additional practical application in providing data to studies on species distribution modeling under future climatic scenarios (Bini et al. 2006), as well as helping onthe-ground conservation measures by the protected area network in the Atlantic Forest of southeastern Brazil.