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Zootaxa 3666 (3): 358 368 www.mapress.com/zootaxa/ Copyright 2013 Magnolia Press Article http://dx.doi.org/10.11646/zootaxa.3666.3.6 http://zoobank.org/urn:lsid:zoobank.org:pub:3cc662f2-2eff-421b-a665-9e4dea621f38 ISSN 1175-5326 (print edition) ZOOTAXA ISSN 1175-5334 (online edition) A new Nototriton (Caudata: Plethodontidae) from Parque Nacional Montaña de Botaderos in northeastern Honduras JOSIAH H. TOWNSEND 1,5, MELISSA MEDINA-FLORES 1,2, ONÁN REYES-CALDERÓN 3 & JAMES D. AUSTIN 4 1 Department of Biology, Indiana University of Pennsylvania, Indiana, Pennsylvania 15705 1081, USA 2 Escuela de Biología, Universidad Nacional Autónoma de Honduras, Tegucigalpa, Francisco Morazán, Honduras 3 Departamento Académico de Estudios Generales, Universidad Nacional de Agricultura, Catacamas, Olancho, Honduras 4 Department of Wildlife Ecology and Conservation, University of Florida, Gainesville, Florida 32611, USA Corresponding author. E-mail: josiah.townsend@iup.edu Abstract The highlands of northeastern Honduras remain under-characterized in terms of biological diversity, as exemplified by the regularity of new amphibian and reptile taxa discoveries. Following the recent description of a new species of Nototriton from the Sierra de Agalta in northeastern Honduras, we report the discovery of a second new species of Nototriton from the nearby Parque Nacional Montaña de Botaderos. This new taxon, Nototriton mime sp. nov., is distinguished from other Nototriton by its distinctive pale brown dorsal coloration in adult males, relatively large nares, a relatively broad head, mitochondrial sequence divergence, and phylogenetic relationships, and is geographically isolated from other populations of Nototriton. Key words: Nototriton mime sp. nov., mtdna, 16S, cytochrome b, Nototriton picucha, sexual dichromatism Resumen Las tierras altas del noreste de Honduras continúan insuficientemente caracterizadas en términos de diversidad biológica, ejemplificado por la regularidad con la que nuevas especies de anfibios y reptiles son descubiertos. Siguiendo la reciente descripción de una nueva especie de Nototriton de la Sierra de Agalta en el noreste de Honduras, reportamos el descubrimiento de una segunda nueva especie de Nototriton, procedente del vecino Parque Nacional Montaña de Botaderos. Este nuevo taxón, Nototriton mime sp. nov., se distingue de otras Nototriton por su distintiva coloración dorsal marrón pálido en machos adultos, orificios nasales relativamente grandes, cabeza relativamente ancha, divergencia en la secuencia mitocondrial y relación filogenética, y se encuentra aislada geográficamente de otras poblaciones de Nototriton. Introduction Despite the presence of over a dozen isolated tropical montane cloud forests, the highlands of northeastern Honduras remain largely uncharacterized in terms of regional endemic biodiversity (Townsend et al. 2011). Only two cloud forest areas above 1,000 m elevation in northeastern Honduras have been studied in any detail: Parque Nacional Sierra de Agalta (Castañeda 2006) and Parque Nacional La Muralla (Espinal et al. 2001), both in Departamento de Olancho. Despite being one of the herpetologically best-known of cloud forests in northeastern Honduras, a new species of moss salamander (Nototriton picucha, Townsend et al. 2011) was recently described from Parque Nacional Sierra de Agalta. The Sierra de Agalta forms the westernmost portion of a narrow cordillera extending into the lowland plain of La Mosquitia and containing the easternmost 2,000+ m elevation cloud forests in Nuclear Central America (Fig. 1). To the north of the Sierra de Agalta, separated by an arid intermontane basin known as the Valle de Agalta and 358 Accepted by Miguel Vences: 24 Apr. 2013; published: 31 May 2013

its bordering xeric pine-oak forest foothills, lies another mountain range, the Sierra de Botaderos (Fig. 1). A relatively large area of intact broadleaf rainforests between 700 and 1,724 m elevation that straddles the border delimiting the departments of Olancho, Colón, and Yoro was recently declared a new protected area, Parque Nacional Montaña de Botaderos (ICF 2011). Previous biological inventory work in the Sierra de Botaderos was limited to two expeditions, one in 2002 and another in 2005, which led to the descriptions of two new pitviper taxa: Atropoides indomitus Smith & Ferrari-Castro (2008) and Cerrophidion wilsoni Jadin, Townsend, Castoe, & Campbell (2012). In April 2011, the authors participated in the first biological inventory of the highest ridge (1,700 1,724 m elevation) in the Sierra de Botaderos. Our team documented herpetofaunal diversity along the targeted ridge on 16 17 April 2011, and collected a series of four Nototriton, the adult males of which had a distinctive yellow-brown dorsal coloration. Analyses of molecular and morphological characteristics confirm that these samples represent a previously unknown species-level taxon that apparently exhibits sexual dimorphism with respect to coloration. We present a new phylogenetic hypothesis for Nototriton based on mitochondrial sequence data and describe this new taxon below. FIGURE 1. Map showing localities mentioned in the text and the distribution of Nototriton in northeastern Honduras. 1) Type locality of N. mime sp. nov., Cerro Ulloa, 1,735 m elevation. 2) Type locality of N. picucha, northwestern slope of Cerro La Picucha, 1,890 m. 3) Type locality of N. lignicola, Cerro de Enmedio, 1,780 m. Method and materials Taxon sampling. Taxa used in this study and their associated metadata are presented in Table 1. Nototriton taxa known to occur in the Chortís Highlands (the region east and south of the tectonic boundary between the Maya and Chortís blocks and north of the Nicaraguan Depression; Townsend 2011) were included in this study, with N. saslaya, the only northern representative of the N. picadoi group (Townsend et al. 2011; Boza-Oviedo et al. 2012), used as an outgroup. Institutional abbreviations follow those standardized by the American Society of Ichthyologists and Herpetologists (Sabaj-Pérez 2012). Specimens were deposited in the Museum of Vertebrate Zoology, University of California Berkeley (MVZ) and the National Museum of Natural History, Smithsonian Institution (USNM). Forest formations follow Holdridge (1967) as applied by McCranie & Wilson (2002). NEW NOTOTRITON FROM NORTHEASTERN HONDURAS Zootaxa 3666 (3) 2013 Magnolia Press 359

TABLE 1. Samples used in sequence divergence and phylogenetic analyses, with GenBank accession and museum voucher numbers; GT = Guatemala, HN = Honduras, NI = Nicaragua. Taxon Locality GenBank voucher GenBank ID 16S cyt b N. barbouri HN: Yoro: Montaña Macuzal UF 156538 GU971733 GU971734 N. brodiei GT: Izabal: Sierra de Caral UTA A-51490 AF199202 AF199139 N. lignicola HN: Olancho: Sierra de La Muralla USNM 497540 AF199204 AF199141 HN: Francisco Morazán: Montaña de Yoro UF 156543 GU971735 GU971736 N. limnospectator HN: Santa Bárbara: San Luís Planes MVZ 263852 JN377383 JQ899197 HN: Comayagua: Azul Meámbar UF 156539 GU971737 GU971738 HN: Comayagua: Azul Meámbar UF 156541 JN377386 JN377396 N. mime sp. nov. HN: Olancho: Sierra de Botaderos USNM 579870 KC905090 KC905094 HN: Olancho: Sierra de Botaderos USNM 579871 KC905091 KC905095 HN: Olancho: Sierra de Botaderos USNM 579872 KC905088 KC905093 HN: Olancho: Sierra de Botaderos MVZ 269306 KC905089 KC905092 N. picucha HN: Olancho: Sierra de Agalta USNM 578299 JN377388 JN377392 HN: Olancho: Sierra de Agalta USNM 578298 JN377389 JN377393 N. sp. A HN: Atlántida: Quebrada de Oro USNM 339712 AF199201 AF199136 N. sp. B HN: Atlántida: Texíguat USNM 578300 JN377387 JN377391 N. stuarti GT: Izabal: Montañas del Mico USAC 3357 JQ899167 JQ899196 N. tomamorum HN: Yoro: Texíguat UF 155377 GU971731 GU971732 N. saslaya NI: Atlantíco Norte: Cerro Saslaya UF 156352 JN377390 JN377394 DNA extraction, PCR amplification, and sequencing. Whole genomic DNA was extracted from tail tips preserved in SED buffer (20% DMSO, 0.25 M EDTA, ph 7.5, NaCl saturated; Seutin et al. 1991, Williams 2007) using the Qiagen PureGene DNA Isolation Kit (Qiagen, Valencia, CA), following manufacturer s instructions. Fragments of the mitochondrial genes 16S large subunit RNA (16S) and cytochrome b (cyt b) were amplified using the primers 16Sar-L and 16Sbr-H for 16S (Palumbi et al. 1991) and MVZ15L and MVZ16H for cyt b (Moritz et al. 1992). PCR reactions were 20mL in total volume, containing ~25ng (~1.8 ml) of DNA template, 4 ml 5X PCR buffer, 1.2 ml MgCl 2 (25mM), 0.09 ml dntps (10 mm), 0.8 ml of each primer (10mM), 0.2 ml GoTaq Flexi polymerase (Promega, Madison, WI, USA), and 11.91 ml H 2 O. Amplification profiles were as follows: 16S, initial denaturation for 3 minutes at 94 C, 35 cycles of denaturation at 94 C for 45 seconds, annealing at 50 C for 45 seconds, and extension at 72 C for 45 seconds, with a final elongation at 72 C for 5 minutes; and cyt b, denaturation for 3 minutes at 94 C, followed by 38 cycles of 94 C for 30 seconds, 48 C for 1 minute, and 1 minute for 45 seconds, and final elongation at 72 C for 5 minutes. Unincorporated nucleotides were eliminated using 1 ul of ExoSAP-IT (USB, Santa Clara, CA, USA) per 10uL of PCR product. We cycle sequenced complimentary strands using the BigDye Terminator 3.1 Cycle Sequencing kit, followed by spin column filtration through Sephadex before electrophoresing on an ABI 3130xl (Applied Biosystems, Inc). Sequence alignment and analyses. Newly generated sequence data for Nototriton from northern Central America were combined with published data available from NCBI (http://www.ncbi.nlm.nih.gov/) for phylogenetic evaluation of Nototriton samples from Montaña de Botaderos, and included trimmed sequences of 490 bp of 16S and 385 bp of cyt b. Sequences were individually aligned for each gene using ClustalW (Thompson et al. 1994) in the program package MEGAv5.05 (Tamura et al. 2011) using the default parameters, adjusted manually, and concatenated, with a final total aligned length of 875 bp. We partitioned the dataset by gene (16S) and by codon position (1st, 2nd, 3rd) for the protein-coding gene cyt b. Best-fit models of nucleotide evolution were selected for each gene and each partition using jmodeltest v2.1 (Posada 2008), which uses PhyML 3.0 (Guindon & Gascuel 2003) for model estimation under a likelihood framework. The number of substitution schemes was set to three to limit the number of models tested to 24, corresponding to the number of different models that can be implemented in MrBayes 3.1.2 (Huelsenbeck & Ronquist 2001). 360 Zootaxa 3666 (3) 2013 Magnolia Press TOWNSEND ET AL.

For both phylogenetic analyses, the dataset was partitioned by gene for 16S and by codon position for cyt b. Bayesian inference (BI) was performed using MrBayes 3.1.2 (Huelsenbeck & Ronquist 2001), and consisted of two parallel runs of four Markov chains (three heated, one cold) run for 20,000,000 generations and sampled every 10,000 generations, with a random starting tree and the first 4,000,000 generations discarded as burn-in, as determined by graphically examining the log likelihood trace plots for stationarity within the program package Geneious Pro R6.1 (Biomatters 2012) The following models were used for BI partitions: 16S GTR+G, cyt b (1st and 2nd codon position) HKY+G, cyt b (3rd codon position) GTR+G. Maximum likelihood (ML) analysis was carried out on the partitioned dataset using RAxML v7.2.8 (Stamatakis 2006; Stamatakis et al. 2008), with 10,000 bootstrap replicates under the default GTR-GAMMA substitution model. Comparative morphology. Measurements were taken with digital calipers, with finer scale measurements taken with a stereo microscope with an optical micrometer; all measurements rounded to the nearest 0.1 mm. Abbreviations used for morphological measurements are as follows: snout to posterior edge of vent, SL; axilla groin length, AG; trunk width at mid-body, TW; head length from tip of snout to gular fold, HL; head width taken at maximum, HW; tail length, TL; hind limb length, HLL; forelimb length, FLL; combined forelimb and hind limb lengths, CLL; hind foot width, HFW; nares length, NL; eye length, EL; eye width, EW; interorbital distance, IOD. To negate the effects of allometric differences and allow for comparative studies across taxa, most measurements are standardized by SL. Specimens examined are listed in the Appendix. Comparative data for other taxa not examined by the authors are taken from Good & Wake (1993), Campbell & Smith (1998), Lynch & Wake (1978), McCranie et al. (1998), Ehmcke & Clemen (2000), Köhler (2002), McCranie & Wilson (2002), Savage (2002), García-Paris & Wake (2000) and Wake & Campbell (2000). Color names and numbers used in the description follow Köhler (2012); descriptions of coloration in life were taken from field notes and a series of photographs of the type specimens. Results Phylogenetic analyses were performed on the concatenated and partitioned 16S/cyt b datasets (Fig. 2), and both BI and ML analyses recovered topologies generally congruent to those in previous studies (Townsend et al. 2010, 2011). Nototriton in the Chortís Highlands are members of six geographically discrete clades (Fig. 2): a northeastern clade (bootstrap score [bs]=98; posterior probability [pp]=1.0), delimited here for the first time, containing N. picucha and four samples from Parque Nacional Montaña de Botaderos; a northern clade (bs=100; pp=1.0), consisting of N. brodiei, N. stuarti, and two unnamed candidate species (N. sp. A and B) presently referred to N. barbouri (sensu lato); a central clade (bs=86; pp=0.99) consisting of N. barbouri (sensu stricto) and N. limnospectator; a N. lignicola clade (bs=100; pp=1.0) and a N. tomamorum clade (single sample), both of whose relationships remain unresolved with respect to the remaining taxa; and N. saslaya (single sample), a member of the otherwise lower Central American N. picadoi clade. Monophyly of the four samples from Parque Nacional Montaña de Botaderos (bs=100, pp=1.0), as well as their sister lineage N. picucha (bs=98, pp=1.0), is unambiguous; one Botaderos sample, MVZ 269306, was excluded from both phylogenetic analyses because the 16S and cyt b sequence fragments were identical to that of USNM 579872. Based on the phylogenetic results, supplemented by morphological data, we herein describe the Parque Nacional Montaña de Botaderos population as a new species. Systematics Nototriton mime sp. nov. Figures 3A D Holotype. USNM 579870 (Fig. 3A), an adult male from Cerro Ulloa (15.3833 N, 86.0399 W), Parque Nacional Montaña de Botaderos, 1,705 m, Departamento de Olancho, Honduras, collected 16 April 2011 by M. Medina- Flores, O. A. Reyes-Calderón, and J. H. Townsend; original field number JHT 3400; GenBank accession numbers KC905090 (16S) and KC905094 (cyt b). NEW NOTOTRITON FROM NORTHEASTERN HONDURAS Zootaxa 3666 (3) 2013 Magnolia Press 361

FIGURE 2. Bayesian phylogram showing phylogenetic relationships of Nototriton mime sp. nov. and other northern Central American species of Nototriton. Maximum likelihood bootstrap scores and Bayesian posterior probability values are shown below branches; bootstrap scores below 50 and posterior probabilities below 0.5 are not shown. Paratopotypes. Three (same collectors as holotype); a female (Fig. 3B; MVZ 269306) and a juvenile (Fig. 3D; USNM 579872), collected 16 April 2011, 1,710 m, 15.3843 N, 86.0396 W; and a male (Fig. 3C; USNM 579871), 17 April 2011, 1,720 m elevation, 15.3874 N, 86.0467 W. Diagnosis. A member of the genus Nototriton diagnosed by possessing 13 costal grooves (>16 costal grooves in Oedipina), hands and feet longer than broad (hands and feet broader than long in Bolitoglossa), and small nares (0.007 0.010 NL/SL; 0.017 0.029 NL/SL in Cryptotriton and Dendrotriton). Distinctiveness of the new species and its assignment to the Nototriton barbouri species group is supported by analysis of sequence fragments from the mitochondrial genes 16S and cyt b (Fig. 2). Nototriton mime is unique among described congeners in the Chortís Highlands by having a yellow-brown dorsal coloration in adult males, as well as relatively longer front and hind limbs (FLL/SL 0.195 0.246, HLL/SL 0.224 0.254; versus 0.142 0.174 and 0.153 0.200 in N. barbouri, 0.148 0.151 and 0.166 0.180 in N. brodiei, 0.137 0.160 and 0.158 0.181 in N. lignicola, 0.156 0.183 and 0.164 0.211 in N. limnospectator, 0.179 0.191 and 0.204 0.218 in N. picucha, 0.172 and 0.178 in N. stuarti, and 0.160 and 0.197 in N. tomamorum). The new species may be further differentiated from the other species of Nototriton found in the Chortís Highlands as follows: from N. brodiei by having a broader head (HW/SL 0.121 0.138, versus 0.120), larger nares (NL/SL 0.007 0.010, versus 0.004 0.005), a shorter tail (TL/SL 0.698 1.117, versus 1.420 1.440), and fewer maxillary teeth (27 46, versus 60 62); from N. lignicola by having a broader head (HW/SL 0.121 0.138, versus 0.103 0.118); from N. limnospectator by having a broader head (HW/SL 0.121 0.138, versus 0.095 0.118) and larger nares (NL/SL 0.007 0.010, versus 0.003); from N. picucha by having a narrower head (HW/SL 0.121 0.138, versus 0.140 0.148); from N. saslaya by having narrower hind feet (HFW/SL 0.044 362 Zootaxa 3666 (3) 2013 Magnolia Press TOWNSEND ET AL.

0.050, versus 0.075 0.091), larger nares (NL/SL 0.007 0.010, versus 0.002 0.003), and a higher number of maxillary (27 46, versus 17 22) and vomerine (12 24, versus 3 11) teeth; from N. stuarti by having a slightly narrower head HW/SL 0.121 0.138, versus 0.138), a shorter tail (TL/SL 0.698 1.117, versus 1.264), and smaller nares (NL/SL 0.007 0.010, versus 0.012); and from N. tomamorum by having well-developed digits with subdigital pads (versus hands and feet syndactylous and lacking subdigital pads), a narrower head (HW/SL 0.121 0.138, versus 0.145), and smaller nares (NL/SL 0.007 0.010, versus 0.018). FIGURE 3. Type series of Nototriton mime sp. nov. A) Male holotype (USNM 579870). Female paratype (MVZ 269306). Male paratype (USNM 579871). Juvenile paratype (USNM 579872). Photographs by JHT. Description of holotype. An adult male (SL = 31.8 mm, total length = 67.3 mm) with a slender body and reduced limbs. The head is rounded, slightly broader than the body; nostrils are relatively small (NL/SL=0.010), NEW NOTOTRITON FROM NORTHEASTERN HONDURAS Zootaxa 3666 (3) 2013 Magnolia Press 363

and the snout is acutely rounded and of moderate length. Nasolabial grooves are visible but slight, and nasolabial protuberances are not apparent. The eyes are relatively large and protuberant, and the elongate parotoid glands are relatively flat and not well-defined. There are 46 maxillary teeth, 3 slightly enlarged premaxillary teeth in line with the maxillaries, and 24 vomerine teeth. The limbs are short (CLL/SL=0.40), with the adpressed limb interval approximately 4.5 costal grooves. The hands and feet are narrow with well-developed digits that bear poorly developed subdigital pads. The relative length of the digits is I<IV<II<III on the hands and I<V<IV II<III on the feet. The tail is longer than the body (TL/SL=1.117) and is very slightly compressed laterally (tail depth 1.04 times tail width at level of cloaca). Measurements of holotype (mm). SL 31.8; AG 16.9; TW 3.3; HL 5.8; HW 3.6; TL 35.5; HLL 6.7; FLL 5.8; CLL 12.5; FFW 0.8; HFW 0.9; NL 0.3; EL 1.8; EW 0.9; IOD 0.7; anterior rim of orbit to snout 1.4; distance separating internal nares 1.1; distance separating external nares 1.6; snout projection beyond mandible 0.3; tip of snout to axilla 9.0; snout to anterior edge of vent 27.8; tail depth at basal constriction 2.7; tail width at basal constriction 2.6. Color in life of adult male holotype. Description of the coloration of the holotype based on a series of color photographs taken in life: dorsal surfaces of paratoid region of head, body, and tail Chamois (84); lateral surfaces mottled Ground Cinnamon (270), with Dark Grayish Brown (284) spots beginning on the centrolateral portion of costal groove 4 and becoming larger posteriorly, with one large blotch present centrolaterally from costal groove 12 to the hind limb; narrow middorsal stripe Pale Pinkish Buff (3), with faint herringbone pattern evident dorsally; dorsal and lateral surfaces of head mottled Vandyke Brown (282) with some Smoky White (261) and Pearl Gray (262) stippling above the mouth; dorsal surface of limbs Sepia (286) with Cinnamon-Rufous (31), Yellow-Ocher (14), and Chamois (84) stippling, becoming more profuse towards body; dorsal surface of tail with Pale Mauve (204) spots beginning about halfway down the length of the tail, with tail becoming almost completely Pale Mauve (204) towards the tip; ventral surface of body with a Dark Mauve (208) to Cyanine Blue (189) cast, with scattered Smoky White (261) and Light Lavender (201) stippling; ventral ground color of tail Fuscous (283), darker than body, with a series of three pairs of Salmon Color (58) blotches behind the vent with spacing equivalent to that of the costal grooves, distal portion of ventral surface of tail with Smoky White (261) and Pale Mauve (204) stippling; color of iris Gem Ruby (65). Variation. In all individuals except the holotype, the tail is shorter than the body (TL/SVL=0.780 in USNM 579871, a male, 0.817 in MVZ 269306, a female, and 0.698 in USNM 579872, a juvenile) In each case, the tail does not appear to be incomplete, and thus the significance of the degree of variation seen in tail length is not clear. The male paratype (USNM 579871) has 27 maxillary, 5 premaxillary, and 12 vomerine teeth; the female paratype (MVZ 269306) has 37 maxillary, 7 premaxillary, and 17 vomerine teeth. The female (MVZ 269306) and juvenile (USNM 579872) paratypes both exhibit a different color pattern than that seen in the male holotype (USNM 579870) and paratype (USNM 579871). The adult female paratype MVZ 269306 has the following coloration in life: dorsal surface of paratoid region of head and body Hazel (26) with darker Brick Red (36) and paler Robin Rufous (29) blotches forming a herringbone pattern, with a narrow Robin Rufous (29) middorsal stripe; irregular Dark Buff Yellow (54) dorsolateral stripes originate approximately halfway down the body and become better defined and continue onto the tail, bordered laterally by a series of small Vandyke Brown (281) blotches positioned on costal grooves 4, 5, 8, 9, and 10, and becoming a continuous blotch over costal grooves 11, 12, and 13 that extends onto the proximal half of the tail; lateral surfaces of body mottled Dark Buff Yellow (54) and Robin Rufous (29) with Pale Bluish Gray (287) stippling; dorsal surface of head with Raw Umber (280) mottling on the parietal region and between the eyes, lateral surfaces of head Russet (44) with Pale Bluish Gray (287) stippling, ventral surface Glaucous (291) with scattered small Pale Bluish Gray (287) spots; ventral surface of body Vandyke Brown (282) with scattered Smoky White (261) and Pale Bluish Gray (287) stippling most prominent near the head and along the centerline of the venter; dorsal surface of tail mottled Hazel (26) and Robin Rufous (29), with Raw Umber (280) to Vandyke Brown (282) lateral and ventral surfaces with Smoky White (261) and Pale Bluish Gray (287) stippling; iris Mahogany Red (34). Etymology. The specific epithet mime is a noun in apposition to the generic name, given to honor our late friend Arquimides Gabriel Rosales Martinez, or Mime (pronounced me-may), a young Honduran biologist passionate about amphibians. Mime and his sister, Novy Hortensia Rosales Martinez, were killed by a drunken driver on 17 December 2010 in Tegucigalpa. Geographic distribution. This species is known only from the immediate vicinity of the type locality, 1,705 364 Zootaxa 3666 (3) 2013 Magnolia Press TOWNSEND ET AL.

1,720 m on Cerro Ulloa, the highest ridge in the Sierra de Botaderos, forming the border between the departments of Colón and Olancho (Figs. 1, 3). FIGURE 4. Type locality of Nototriton mime sp. nov. A) Cerro Ulloa (center), highest point 1,724 m elevation, Parque Nacional Montaña de Botaderos; note the recent and ongoing deforestation on the approaching slopes. B) Tropical montane cloud forest along summit ridge of Cerro Ulloa, ca. 1,720 m elevation. Photographs by JHT. NEW NOTOTRITON FROM NORTHEASTERN HONDURAS Zootaxa 3666 (3) 2013 Magnolia Press 365

Natural history. The habitat at the type locality is tropical montane cloud forest, in the Lower Montane Wet Forest formation. The male holotype (USNM 579870; Fig. 3A) was active at night among the leaves of an orchid growing on a small stump on the ground along a wet ridge at 1,705 m elevation. The male paratype (USNM 579871; Fig. 3C) was collected from underneath a small log embedded in the ground at 1,720 m elevation. The female paratype (MVZ 269306; Fig. 3B) was found in a bromeliad on the ground at 1,705 m elevation, near the holotype. The juvenile paratype (USNM 579872; Fig. 3D) was collected during the late afternoon from within a small bromeliad approximately 2 m above the ground on a large fallen branch at 1,710 m elevation. Conservation status. Based on the extremely limited extent of intact cloud forest around the only known locality (approximately 300 hectares), and the continuing deforestation surrounding this locality, we recommend Nototriton mime be immediately considered Critically Endangered (CR B1ab[i, ii, iii]+2ab[i, ii, iii]), following the IUCN Red List Criteria (IUCN 2012). The broadleaf forests of the Sierra de Botaderos are approximately 33,000 hectares in total extent, and ranges from 100 to 1,724 m elevation. However, less than 1,000 total hectares of forest above 1,500 m elevation exists, and is fragmented into approximately a dozen discrete patches along the spine of the cordillera. The type locality, Cerro Ulloa, is one of these patches, and is the highest elevation forest in the mountain range. Cerro Ulloa is already deforested up to and above 1,500 m elevation on the southern slopes (Fig. 4A), and preexisting trails made by farmers cross the summit of the peak (Fig. 4B) and access inholdings deep with the forest on the Colón side of the ridge. The approximately 300 hectares of intact cloud forest habitat surrounding the type locality are already under direct threat from migratory agriculture and cattle ranching, and, most recently, are indirectly threatened by the dramatic increase in international narcotrafficking in adjacent portions of Colón and Olancho. The recent declaration of the remaining premontane and lower montane rainforests as a national park is a positive development; however, the remote location of the park and high-risk nature of conflicts related to enforcing protection of its remaining forests make conservation here, and throughout eastern Honduras, a resourceintensive challenge. Remarks. With the description of Nototriton mime, there are 17 named species of Nototriton distributed from Guatemala to Costa Rica, six of which are endemic to Honduras (Townsend et al. 2011; Boza-Oviedo et al. 2012). Each of these six endemic species have distributions restricted to cloud forest habitat in the upper reaches of isolated mountain ranges, with only one species (N. limnospectator) known to occur at premontane elevations as low as 800 m. In addition to these six species, there are two as yet undescribed species currently referred to the taxon N. barbouri, one from Parque Nacional Pico Bonito (N. sp A in Townsend et al. 2011) and one from Refugio de Vida Silvestre Texíguat (N. sp. B, op. cit.). The coloration seen in Nototriton mime appears to be the first documented case of sexual dichromatism in the genus Nototriton, and one of few documented cases in tropical salamanders. Another endemic Honduras salamander, Bolitoglossa diaphora, from the Sierra de Omoa in northwestern Honduras, also exhibits marked dichromatism, with males being bluish-gray and females being a mottled red-orange coloration (McCranie & Wilson 1995). Acknowledgments Research in Honduras was carried out under permits issued to JHT by the Instituto Nacional de Conservación y Desarrollo Forestal, Áreas Protegidas y Vida Silvestre [ICF] (Resolución DE-MP-086-2010 and Dictamen DVS- ICF-045-2010); we thank S. Laínez, I. Acosta, and R. Downing for assistance in obtaining these permits. Fieldwork in Parque Nacional Montaña de Botaderos was undertaken as part of an Explorer s Club expedition (Flag # 93), which was organized by M. Bonta and supported financially by R. Hyman. M. Bonta, R. Hyman, D. Medina, F. Steiner, and R. Ulloa also participated in the expedition. M. Bonta and W. Brenneman provided comments on a preliminary draft of this manuscript, which was further improved by peer reviews from A. Hertz and L.D. Wilson. I.R. Luque-Montes provided the map used in Figure 1. References Biomatters (2012) Geneious version R6.1. Available from: http://www.geneious.com/ (Accessed 7 May 2013) Boza-Oviedo, E., Rovito, S.E., Chaves, G., García-Rodríguez, A., Artavia, L.G., Bolaños, F. & Wake, D.B. (2012) Salamanders from 366 Zootaxa 3666 (3) 2013 Magnolia Press TOWNSEND ET AL.

the eastern Cordillera de Talamanca, Costa Rica, with descriptions of five new species (Plethodontidae: Bolitoglossa, Nototriton and Oedipina) and natural history notes from recent expeditions. Zootaxa, 3309, 36-61. Campbell, J.A. & Smith, E.N. (1998) New species of Nototriton (Caudata: Plethodontidae) from eastern Guatemala. Scientific Papers of the Natural History Museum of the University of Kansas, 6, 1 8. Castañeda, F.E. (2006) Herpetofauna del Parque Nacional Sierra de Agalta, Honduras. International Resources Group, United States Agency for International Development, Washington, D.C., 65 pp. Ehmcke, J. & Clemen, G. (2000) Teeth and their sex-dependent dimorphic shape in three species of Costa Rican plethodontid salamanders (Amphibia: Urodela). Annals of Anatomy 182, 403 414. http://dx.doi.org/10.1016/s0940-9602(00)80045-x Espinal, M.R., McCranie, J.R. & Wilson, L.D. (2001) The herpetofauna of Parque Nacional La Muralla, Honduras. In: Johnson, J.D., Webb, R.G., & Flores-Villela, O.A. (Eds.), Mesoamerican Herpetology: Systematics, Zoogeography and Conservation. The University of Texas, El Paso, pp. 100 108. García-París, M. & Wake, D.B. (2000) Molecular phylogenetic analysis of relationships of the tropical salamander genera Oedipina and Nototriton, with descriptions of a new genus and three new species. Copeia, 2000, 42 70. http://dx.doi.org/10.1643/0045-8511(2000)2000[0042:mpaoro]2.0.co;2 Good, D.A. & Wake, D.B. (1993) Systematic studies of the Costa Rican moss salamanders, genus Nototriton, with descriptions of three new species. Herpetological Monographs, 7, 131 159. http://dx.doi.org/10.2307/1466956 Guindon, S. & Gascuel, O. (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology, 52, 696 704. http://dx.doi.org/10.1080/10635150390235520 Holdridge, L.R. (1967) Life Zone Ecology. Revised Edition. Tropical Science Center, San José, Costa Rica. 206 pp. Huelsenbeck, J.P. & Ronquist, F.R. (2001) MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics (Oxford), 17, 754 755. http://dx.doi.org/10.1093/bioinformatics/17.8.754 ICF (2011) Aviso de Declaratoria del Área Protegida del Parque Nacional Montaña de Botaderos. Instituto Nacional de Conservación y Desarollo forestal, Áreas Protegidas, y Vida Silvestre, Comayagüela, Honduras. Available from: http://www.icf.gob.hn/secciones/noticias.aspx?id=17a3eb44-0a0f-4c9f-8dd4-2652c11a8d90 (Accessed 13 February 2013) IUCN (2012) IUCN Red List Categories and Criteria: Version 3.1. Second Edition. IUCN, Gland, Switzerland and Cambridge, United Kingdom. Available from: http://www.iucnredlist.org/static/categories_criteria_3_1 (Accessed 13 February 2013) Jadin, R.C., Townsend, J.H., Castoe, T.A. & Campbell, J.A. (2012) Cryptic diversity in disjunct populations of Middle American Montane Pitvipers: a systematic reassessment of Cerrophidion godmani. Zoologica Scripta, 41, 455-470. http://dx.doi.org/10.1111/j.1463-6409.2012.00547.x Köhler, G. (2002) A new species of salamander of the genus Nototriton from Nicaragua (Amphibia: Caudata: Plethodontidae). Herpetologica, 58, 205 210. http://dx.doi.org/10.1655/0018-0831(2002)058[0205:ansoso]2.0.co;2 Köhler, G. (2012) Color Catalogue for Field Biologists. Herpeton Verlag, Offenbach, Germany, 49 pp. Lynch, J.F. & Wake, D.B. (1978) A new species of Chiropterotriton (Amphibia: Caudata) from Baja Verapaz, Guatemala, with comments on relationships among Central American members of the genus. Contributions in Science, Natural History Museum of Los Angeles County, 294, 1 22. McCranie, J.R. & Wilson, L.D. (1995) A new species of salamander of the genus Bolitoglossa (Caudata: Plethodontidae) from Parque Nacional El Cusuco, Honduras. Journal of Herpetology, 29, 447 452. http://dx.doi.org/10.2307/1564996 McCranie, J.R. & Wilson, L.D. (2002) The Amphibians of Honduras. Society for the Study of Amphibians and Reptiles, Contributions in Herpetology. 19. Ithaca, New York. i x, 1 625. McCranie, J.R., Wilson, L.D. & Polisar, J. (1998) Another new montane salamander (Amphibia: Caudata: Plethodontidae) from Parque Nacional Santa Barbara, Honduras. Herpetologica, 54, 455 461. Moritz, C., Schneider, C.J. & Wake, D.B. (1992) Evolutionary relationships within the Ensatina eschscholtzii complex confirm the ring species interpretation. Systematic Biology, 41, 273 291. Palumbi, S.R., Martin, A.P., Romano, S., McMillan, W.O., Stice, L. & Grabowski, G. (1991) The Simple Fool s Guide to PCR. Special Publication of the Department of Zoology. University of Hawaii, Honolulu, 45 pp. Posada, D. (2008) jmodeltest: phylogenetic model averaging. Molecular Biology and Evolution, 25, 1253 1256. http://dx.doi.org/10.1093/molbev/msn083 Sabaj-Pérez, M.H. (Ed.) (2012) Standard symbolic codes for institutional resource collections in herpetology and ichthyology: an Online Reference. Version 3.0 (23 February 2012). American Society of Ichthyologists and Herpetologists, Washington, D.C., Available from: http://www.asih.org/ (Accessed 13 February 2013) Savage, J.M. (2002) The Amphibians and Reptiles of Costa Rica. A Herpetofauna between Two Continents, Between Two Seas. Cornell University Press, Ithaca, New York. 934 pp. Seutin, G., White, B.N. & Boag, P.T. (1991) Preservation of avian blood and tissue samples for DNA analyses. Canadian Journal of Zoology, 69, 82 90. http://dx.doi.org/10.1139/z91-013 Smith, E.N. & Ferrari-Castro, J.A. (2008) A new species of jumping pitviper of the genus Atropoides (Serpentes: Viperidae: Crotalinae) from the Sierra de Botaderos and the Sierra La Muralla, Honduras. Zootaxa, 1948, 57 68. NEW NOTOTRITON FROM NORTHEASTERN HONDURAS Zootaxa 3666 (3) 2013 Magnolia Press 367

Stamatakis, A. (2006) RAxML-VI-HPC: Maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics, 22, 2688 2690. http://dx.doi.org/10.1093/bioinformatics/btl446 Stamatakis, A., Hoover, P. & Rougemont, J. (2008) A rapid bootstrap algorithm for the RAxML web-servers. Systematic Biology, 75, 758 771. http://dx.doi.org/10.1080/10635150802429642 Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731 2739. http://dx.doi.org/10.1093/molbev/msr121 Thompson, J.D., Higgins, D.G. & Gibson, T.J. (1994) CLUSTALW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22, 4673-4680. http://dx.doi.org/10.1093/nar/22.22.4673 Townsend, J.H. (2011) Integrative Taxonomy Reveals the Chortís Block of Central America as an Underestimated Hotspot of Amphibian Diversity. Unpublished PhD Dissertation, University of Florida, Gainesville, Florida. 350 pp. Townsend, J.H., Butler, J.M., Wilson, L.D. & Austin, J.D. (2010) A distinctive new species of moss salamander (Caudata: Plethodontidae: Nototriton) from an imperiled Honduran endemism hotspot. Zootaxa, 2434, 1 16. Townsend, J.H., Medina-Flores, M., Murillo, J.L. & Austin, J.D. (2011) Cryptic diversity in Chortís Highland moss salamanders (Caudata: Plethodontidae: Nototriton) revealed using mtdna barcodes and phylogenetics, with a new species from eastern Honduras. Systematics and Biodiversity, 9, 275 287. http://dx.doi.org/10.1080/14772000.2011.611188 Wake, D.B. & Campbell, J.A. (2000) A new species of diminutive salamander (Amphibia: Caudata: Plethodontidae: Nototriton) from the Montañas del Mico of Guatemala. Proceedings of the Biological Society of Washington, 113, 815 819. Williams, S.T. (2007) Safe and legal shipment of tissue samples: does it affect DNA quality? Journal of Molluscan Studies, 73, 416 418. http://dx.doi.org/10.1093/mollus/eym039 APPENDIX. Comparative specimens examined (49). Nototriton barbouri (16; including N. sp. A and N. sp. B) HONDURAS: Atlántida USNM 339712, 497552, 578300; Yoro UF 156538, USNM 339700 11. Nototriton lignicola (16) HONDURAS: Francisco Morazán UF 156542 44; Olancho: USNM 497539 51. Nototriton limnospectator (13) HONDURAS: Comayagua UF 156539 41; Cortés IRL 035 (in UF collection, May 2009), USNM 573715 17; Santa Bárbara UF 98461 66. Nototriton picucha (2) HONDURAS: Olancho USNM 578298 99. Nototriton saslaya (1) NICARAGUA: Jinotega UF 156532. Nototriton tomamorum (1) HONDURAS: Yoro UF 155377. 368 Zootaxa 3666 (3) 2013 Magnolia Press TOWNSEND ET AL.