First survey of the amphibians and reptiles of the Nectandra Cloud Forest Reserve , Alajuela , Costa Rica

We conducted a first survey of the amphibians and reptiles of the Nectandra Cloud Forest Reserve in June 2010, during the rainy season in Costa Rica. We found a total of 30 species of amphibians and reptiles in 15 families during our weeklong survey, including 15 frogs, 2 salamanders, 7 lizards, and 6 snakes. We compare our results to those of a previous survey of the nearby San Ramón Reserve.


INTRODUCTION
Despite its relatively small area (51,100 km 2 ), Costa Rica is rich in amphibians and reptiles.One hundred and ninetynine species of amphibians (AmphibiaWeb 2014) and 241 species of reptiles (F. Bolaños, pers. comm.) are reported from the country, and new species continue to be described (García-París et al. 2008;Chaves et al. 2009;Boza-Oviedo et al. 2012).While some parts of the country are well known biologically, especially reserves such as La Selva (Guyer and Donnelly 2004) in the northern lowlands, Monteverde Cloud Forest Reserve in the Cordillera de Tilarán (Hayes et al. 1989), Reserva Biológica Alberto ML Brenes near San Ramón (Bolaños and Ehmcke 1996), Rincón de Osa on the Corcovado Peninsula (Scott 1976), and San Vito in the Fila Costeña (Santos-Barrera et al. 2008), other areas of the country have received less attention.We report the results of a weeklong survey of the Nectandra Reserve, in Alajuela province.This was the first study of the herpetofauna of the reserve.

Study site
The Nectandra Reserve is located north of San Ramón on the highway to Fortuna (10.18564°N, 084.50787°W,WGS84 datum), across the continental divide from the well-known Monteverde Cloud Forest Reserve (Figure 1) in the Province of Alajuela, Costa Rica.The reserve is east of the continental divide (Caribbean drainage) in the southeast portion of the Cordillera de Tilarán, and has an extent of 130 ha.Elevations on the reserve range from approximately 1,000-1,200 m, and the forest is classified as premontane rainforest (Holdridge 1967).Within or immediately adjacent to the reserve, habitats include primary forest, secondary forest, abandoned cacao plantation, and cattle pasture.The reserve contains several streams as well as some small man-made ponds adjacent to the entrance.

Data collection
We conducted a first survey of the Nectandra Reserve from 10-16 June 2010 in order to document the herpetofauna of the reserve and establish a baseline for future biological work and management.The survey was conducted during the rainy season, when amphibian species are most active.From 10-14 June, all authors were involved in survey work, with help from one local guide at all times, while only the first author (S.Rovito) conducted surveys from 15-16 June.We surveyed opportunistically by walking trails during the day and searching for amphibians and reptiles on vegetation, under logs, in leaf litter, and along streams.We also walked along the highway that borders the reserve to the east in order to collect road-killed snakes.By night, we searched along trails and streams with headlamps.We estimate that we searched for 4-6 hours each day and 1-3 hours per night.Because no herpetological collection yet exists from the reserve, we collected up to three specimens of each species.A permit for specimen collection was issued by MINAET (089 2010-SINAC).Amphibians and reptiles were euthanized using standard methods, preserved in 10% buffered formalin and stored in 95% ethanol.Tissues of each specimen were collected in RNAlater for future genetic analyses.All specimens and tissues were deposited in the herpetological collection of the Museo de Zoología, Universidad de Costa Rica (Tables 1 and 2 for specimen numbers).Species identifications were assigned in the field using keys and information provided in Savage (2002), Köhler (2008), and Köhler (2011) and confirmed or updated by Federico Bolaños (Universidad de Costa Rica).We followed the taxonomy of AmphibiaWeb ( 2014) for all amphibian species identifications and that of Köhler (2008) for reptile species.

DISCUSSION
The amphibians and reptiles that we found mainly represent a subset of those known from the more extensively surveyed Reserva Biológica Alberto ML Brenes, San Ramón (hereafter Reserva San Ramón).Bolaños and Ehmcke (1996) reported 30 amphibian and 35 reptile species from the Reserva San Ramón.They listed as common the same species of direct developing terrestrial frogs and Anolis that we found to be abundant at Nectandra.Their list includes a number of species that we did not find, including frogs such as Atelopus varius (Lichtenstein & Martens, 1856) and Craugastor andi (Savage, 1974) that have experienced severe declines and extirpations (Pounds and Crump 1994;Pounds et al. 1997).Bolaños and Ehmcke (1996) list Craugastor crassidigitus as uncommon and in decline, while we found it to be  relatively common.They also reported species, such as Ecnomiohyla miliaria (Cope, 1886) and Anotheca spinosa (Steindachner, 1864) that are difficult to detect and may be present at Nectandra but would not be expected to be found during a short survey.
One notable species we found at Nectandra is the salamander Nototriton gamezi García-París & Wake 2000.This species is known from only two localities in Costa Rica: the type locality in the Monteverde Cloud Forest Reserva (García-París and Wake 2000) and the Reserva San Ramón (reported as N. abscondens by Bolaños and Ehmcke (1996)).The Nectandra Reserve is located between these two reserves, and adds a third known site to the distribution on this narrowly endemic species.We found three species of amphibians (Isthmohyla pseudopuma, Oedipina uniformis, Teratohyla pulverata) and one species of reptile (Lepidoblepharis xanthostigma) that were not reported by Bolaños and Ehmcke (1996).
The survey of the Reserva San Ramón was conducted at elevations of 800-1,200 m, approximately the same altitudinal range as our survey of Nectandra.Because of the close geographic proximity of the two reserves and their similarity in habitats and elevation, we expect the list of amphibians and reptiles from Nectandra will more closely resemble the more complete list from the Reserva San Ramón if more extensive fieldwork were to take place at Nectandra.The relatively large number of species of amphibians and reptiles found at Nectandra, including several not reported from a previous survey of the Reserva San Ramón, shows that Nectandra is an important component of the system of protected areas linking the Cordillera de Tilarán and the Cordillera Central of Costa Rica.Amphibian declines have been reported at sites in both of these regions (including Monteverde and the Reserva San Ramón) and having current information on amphibian populations at other sites is important for understanding these declines in a regional and temporal context.The elevation of Nectandra places it in the same range as sites where major amphibian declines have occurred in Costa Rica (Pounds and Crump 1994;Young et al. 2001), and the results of this survey can be used as an additional reference for the status of post-decline amphibian populations.Nectranda is a site that could be used in plans for the recovery of surviving species that suffered declines.Additional surveys of the herpetofauna of Nectandra should be conducted in order to find rare or elusive species that were not seen during our short survey.The reserve also represents a promising site for scientific research and public outreach efforts, given its easy access and high biological diversity.

Figure 1 .
Figure 1.Map of Costa Rica, showing locations of areas discussed in the text where amphibian and reptile surveys have been conducted (blue squares).

Table 2 .
Reptile species found in the Nectandra Cloud Forest Reserve, and associated voucher numbers for specimens deposited at the University of Costa Rica.