Systematics of the Rhinellamargaritifera complex (Anura, Bufonidae) from western Ecuador and Panama with insights in the biogeography of Rhinellaalata Author dos Santos, Sueny P. Author Ibanez, Roberto Author Ron, Santiago R. text ZooKeys 2015 501 109 145 http://dx.doi.org/10.3897/zookeys.501.8604 journal article http://dx.doi.org/10.3897/zookeys.501.8604 1313-2970-501-109 CBFCB838C07B4329AEEAF61F8F12C16C Taxon classification Animalia Anura Bufonidae Rhinella alata (Thominot, 1884) Bufo alatus Thominot, 1884. Holotype: MNHN 84285, adult male from Obispo, Panama. Diagnosis. Rhinella alata is a small-sized (Table 2; Figs 8 and 9) species of Rhinella having the following combination of characters: (1) average SVL of females 44.25 mm (SD = 4.36, n = 39), males 36.83 mm (SD = 2.31, n = 49); (2) bony knob at angle of jaws absent, corner of mouth angular; (3) supraorbital crests low and thick, continuous with preorbital crests; usually with crenulate texture on vertical surfaces; (4) supratympanic crests concave and small; their posterior edge usually next to the anterior border of parotoid glands; (5) canthus rostralis present but inconspicuous, sometimes continuous with preorbital crests; (6) parietal crests usually present, ill-defined; (7) heel reaching posterior margin of eye when hindlimbs adpressed; (8) vertebral apophyses no protruding; (9) snout subacuminate in dorsal view, from rounded to protruding in profile; (10) skin on dorsum bearing a mixture of warts, pustules, and minute tubercles; (11) mid-dorsal line from snout to vent often present; (12) spiculate tubercles on external border of shank, evident especially on females; (13) dorsolateral row of sharply pointed, conical tubercles between posterior border of parotoid glands and groin; (14) tympanic membrane and tympanic annulus distinct; moderately large, ovoid to round; (15) parotoid glands small, elongated posteriorly; (16) upper eyelid warty; (17) tarsal fold absent; (18) digits slender and long, with small knobs at tip; lateral fringes present; finger lengths 3> 4> 2> 1; toe lengths 4> 5> 3> 2> 1; (19) nuptial pads present. Rhinella alata is most similar to Rhinella acutirostris . Both species differ from other members of the Rhinella margaritifera group by the absence of protruding vertebral apophyses, canthus rostralis not raised, snout projected, and low cranial crests. Rhinella acutirostris differs from Rhinella alata in having a bony knob at the angle of jaws (bony knob absent in Rhinella alata [ Hoogmoed 1986 , Loetters and Koehler 2000 ]). Rhinella alata differs from the holotype of Rhinella proboscidea (ZSM 1145/0) in having a less protruding snout and skin on dorsum bearing a mixture of warts, pustules, and minute tubercles (smooth skin in Rhinella proboscidea ). Rhinella dapsilis is much larger than Rhinella alata ( Rhinella dapsilis holotype SVL = 77 mm, adult male; Myers and Carvalho 1945 ) and has a fleshy proboscis in the snout (proboscis absent in Rhinella alata ). Rhinella alata differs from Rhinella yunga in having tympanic membrane and annulus distinct (tympanic membrane and annulus absent in Rhinella yunga ; Moravec et al. 2014 ). Rhinella hoogmoedi , Rhinella magnussoni , Rhinella martyi , Rhinella paraguayensis , Rhinella scitula , Rhinella sclerocephala , and Rhinella stanlaii have a bony knob at angle of jaws ( Caramaschi and Pombal 2006 , Lima et al. 2007 , Fouquet et al. 2007a , Avila et al. 2010 , Caramaschi and Niemeyer 2003 , Mijares-Arrutia and Arends-R 2001 , Loetters and Koehler 2000 ; bony knob absent in Rhinella alata ). Rhinella alata differs from Rhinella castaneotica , Rhinella margaritifera (sensu stricto) and Rhinella roqueana , by the absence of protruding vertebral apophyses (present in Rhinella castaneotica [ Caldwell 1991 ], Rhinella margaritifera [ Lavilla et al. 2013 ], and Rhinella roqueana [ Melin 1941 ]). Rhinella alata is most closely related to populations of Rhinella margaritifera from the upper Amazon basin in Ecuador and Peru. They can be easily distinguished by differences in body size (Fig. 5; see morphometric comparisons section) and relative size of cranial crests (Fig. 6). Holotype. The holotype is an adult male with SVL = 39.2 mm (Fig. 11). Descriptions of the holotype have been provided by Leavitt (1933) and Hoogmoed (1989) . The bony knob at angle of jaws and vertebral apophyses are absent. The crests are low and thick. There is a dorsolateral row of conical tubercles from the posterior border of the parotoid gland to the groin. There is a clear mid-dorsal line from the snout to the vent. The tympanum is rounded. Variation. Variation in dorsal and ventral coloration of preserved specimens is shown in Figures 12 and 13. Background dorsal coloration varies from light gray (QCAZ 37244, AMNH 88689), light brown (QCAZ 14607, AMNH 104454) to dark gray (QCAZ 6733) or dark brown (QCAZ 11598, AMNH 52744), with irregular black and yellowish marks (QCAZ 4444, AMNH 88690). Some specimens have nearly uniform brown dorsum without marks (QCAZ 31603, 10296, AMNH 10296). A clear mid-dorsal line is often present (e.g. QCAZ 3502, QCAZ 12233). Figure 12. Rhinella alata from Ecuador showing variation in dorsal and ventral coloration of preserved specimens. Left to right, males: QCAZ 6733 (SVL 38.23 mm), QCAZ 10279 (SVL 35.08 mm); females, QCAZ 11598 (SVL 42.13 mm), QCAZ 14607 (SVL 50.95 mm), QCAZ 10439 (SVL 47.06 mm). See Appendix 1 for locality data. Not shown at the same scale. Figure 13. Rhinella alata from Panama showing variation in dorsal and ventral coloration of preserved specimens. Left to right, male: AMNH 89459 (SVL 37.54 mm); females, AMNH 88694 (SVL 41.21 mm), AMNH 55476 (SVL 41.19 mm), AMNH 104454 (SVL 49.69 mm), AMNH 88689 (SVL 42.75 mm), AMNH 20896 (SVL 42.98 mm). See Appendix 1 for locality data. Not shown at the same scale. Ventral surfaces of preserved specimens have a cream to yellowish-cream background color with irregular darker marks arranged in diverse patterns; marks can be light gray (QCAZ 6734, AMNH 88689), light brown (QCAZ 6732, AMNH 104454), dark gray (QCAZ 31606) or dark brown (QCAZ 6733, AMNH 89459), and vary from being restricted to the anterior half of the body (QCAZ 31604, AMNH 89459) to being present over the entire venter (QCAZ 4445, AMNH 88694). A longitudinal mid-ventral cream thin stripe can be present in the gular region (QCAZ 31602, 31606) or from the gular region to the mid-venter (QCAZ 6731, 11598). Head shape in dorsal view varies from elongated (QCAZ 11598, AMNH 89459) to subtriangular (QCAZ 4447, AMNH 55475); in lateral view it varies from rounded (QCAZ 31605, AMNH 52749) to protruding (QCAZ 11393, AMNH 55475). Canthal region coloration varies from light gray or light brown to dark gray or dark brown. In some individuals the area below the eye and tympanum is yellowish cream (QCAZ 4447, AMNH 20896) or brown (QCAZ 31603, AMNH 88694) and differs from the color of the dorsum. Cloacal tubercles vary from yellowish cream (QCAZ 4441, AMNH 20896), to gray (QCAZ 31606) or brown (QCAZ 31602, AMNH 88695). Color in life. Based on digital photograph of an adult female QCAZ 50568 (Fig. 8). Dark brown dorsum with irregular light brown and yellowish marks; there is a clear mid-dorsal line. Dorsal surfaces of tights and shanks are dark brown with transversal brown bands. Dorsal surfaces of forelimbs are dark brown with irregular light brown marks. Dark brown tubercles are abundant on the dorsum. Ventral surfaces vary from light brown to dark brown, with some irregularly distributed white and orange spots. The fingertips and the subarticular tubercles on fingers and toes are red-orange. Canthal region and tympanum are dark brown; iris greenish yellow with black reticulation. Based on a digital photography of an adult male QCAZ 37248 (Fig. 8). Light brown dorsum with black spots and light brown and light gray marks. Dorsal surfaces of tights, shanks and forelimbs are light brown with transversal dark brown bands. Brown tubercles are abundant on the dorsum. Ventral surfaces are dark brown with irregularly distributed yellowish marks; the posterior part of the venter is cream. The subarticular tubercles of palms, soles, and fingertips are red-orange. Canthal region and tympanum are dark brown; iris greenish yellow with black reticulation. Distribution and ecology. Rhinella alata has been recorded at 37 localities in the Ecuadorian Choco ( Canar , Carchi, El Oro, Esmeraldas, Manabi , Pichincha, and Santo Domingo Provinces; Fig. 1), one locality in the Colombian Choco (Barbacoas, Narino ; see Taxonomic remarks) and 35 localities in Panama (Comarca Guna Yala and Provinces Cocle , Colon , Darien and Panama; Fig. 2). It has a wide elevation range, from 19 to 1500 m above sea level. The examined specimens from Chocoan populations contain 21 gravid females (average SVL = 45.37 mm, SD = 4.05 mm): QCAZ 4262, QCAZ 4441, QCAZ 4442, QCAZ 4443, QCAZ 7065, QCAZ 10296, QCAZ 11597, QCAZ 11598 collected in January; QCAZ 50568 collected in February; QCAZ 11392, QCAZ 31601, QCAZ 31603, QCAZ 31605 collected in April; QCAZ 25023 collected in June; QCAZ 10439 collected in August; QCAZ 14607 collected in November; QCAZ 10301 collected in December. This suggests year round reproductive activity with a peak between January and April, a period that corresponds to the rainy season in the Ecuadorian Choco . In Panamanian populations gravid females were found in January (AMNH 104454), September (AMNH 55461), November (AMNH 88689), and December (AMNH 53699). In central Panama, Rhinella alata breeds in ponds and pools along permanent streams or swamps. Reproduction is explosive and most takes place from the middle of the rainy season to early dry season ( Wells 1979 , Ibanez et al. 1999 ). Choruses last less than 24 hours with males usually calling at night and oviposition occurring by day, especially in the early afternoon ( Wells 1979 ). Otherwise, individuals are primarily diurnal, found active on the leaf litter of the forest floor during daytime, and often found asleep on leaves of low vegetation at night ( Ibanez et al. 1999 ). Diet is specialized on ants ( Toft 1981 ). Most of the Ecuadorian specimens are from Reserva Mayronga and Reserva Ecologica Cotacachi-Cayapas. They were found in the leaf litter of secondary forest and in agricultural lands. Some adults were observed at night within the forest in vegetation above the ground and some were found in amplexus (QCAZ 10271, QCAZ 10274, QCAZ 10275 in November 1996, and QCAZ 31604, QCAZ 31605 in February 1996). All the specimens collected in Reserva Ecologica Cotacachi-Cayapas were found in secondary forest. At some collecting sites, the forest has been cleared for cacao plantations (QCAZ specimen database). According to the classification of Sierra et al. (1999) the vegetation types for Ecuadorian localities are: (1) Lowland Evergreen Forest of Coastal Range, characterized by abundant epiphytes, climbers and herbaceous plants, with a canopy of 30 m (e.g. Reserva La Chiquita, Durango); (2) Semideciduous Lowland Forest of Coastal Range, defined by the presence of broad canopy trees up to 20 m and curved shafts; the tree stratum is characterized by the presence of spiny, deciduous species with epiphytes while the forest floor has herbaceous plants (e.g. Bilsa, La Tortuga); (3) Evergreen Foothill Forest of Coastal Range, characterized by a canopy that can reach 30 m or more and trunks of trees covered with orchids, bromeliads, ferns and aroids (e.g. Manta Real, Alto Tambo); (4) Deciduous Lowland Forest of Costal Range, characterized by losing leaves during part of the year with a great varieties of cactus and thorny plants; the most conspicuous trees are the family Bombacaceae have curved trunks and broad crown. (e.g. El Progreso); (5) Semideciduos Foothill Forest of Coastal Range, characterized by having slightly dispersed vegetation, with trees over 20 m and dense herbaceous layers of ferns (e.g. Valle Hermoso). The main vegetation types for Panamanian localities are (following Hogan 2010 ): (1) Isthmian-Atlantic Moist Forests, characterized by tall tropical evergreen forest with buttressed canopy trees reaching 40 m and with an extremely rich epiphyte flora (e.g. Cruces Trail, Punta Rincon ); (2) Eastern Panamanian Montane Forest, at elevations from 500 to 1800 m above sea level, includes marshes, swamp forests, semi-deciduous tropical moist forests, premontane wet forest, cloud forests and elfin forests (e.g. Cana, Cerro Tacarcuna); (3) Choco-Darien Moist Forests, at elevations between 0 and 1000 m above sea level, between the Pacific Ocean and the western range of the Andes (e.g. Dad Nakue Dubpir, Udirbi). Taxonomic remarks. Based on morphological characters, Velez-Rodriguez (2004) ascribed four populations from Panama and Colombia to Rhinella alata : Isthmus of Panama (Panama; 15 males, 10 females); Parque Nacional Los Katios (Colombia; 12 males, 15 females); Gorgona and Gueape Island (Colombia; 7 males, 8 females); Municipio Restrepo (Colombia; 7 males, 8 females). Based on data from Velez-Rodriguez (2004) , these populations differ from the holotype of Rhinella alata and populations of Rhinella alata in Ecuador and Panama (in parentheses) in having: (1) a canthus rostralis protruding in females and ill-defined in males (inconspicuous in males and females), (2) parietal crests well defined in females, ill-defined in males (ill-defined in males and females), (3) vertebral apophyses slightly visible externally (absent). The differences suggest that those specimens are not Rhinella alata and may belong to a different species. Alternatively, differences between Rhinella alata described by Velez-Rodriguez (2004) and our study could be an artifact resulting from the use of distinct terminology for similar character states. In contrast, Mueses-Cisneros and Moreno-Quintero (2012) reported two species of the Rhinella margaritifera group form Barbacoas, Narino , Colombia ( Rhinella sp. 9 and Rhinella sp. 10). Two photographs of live individuals (pp. 45) show morphological features that fall within the observed variation of Rhinella alata . We tentatively assign them to Rhinella alata but direct specimen examination is required to confirm this identification.