Bioacoustics of Ameerega shihuemoy

Distribution and natural history. Ameerega shihuemoy is distributed in southeastern Peru at the transition between the montane forest and the lowlands. Its altitudinal range spans from 340 to 850 m above sea level. This species is known from nine localities; three in the buffer zone of the Manu Biosphere Reserve (The Manu Learning Centre, Erika lodge, and Aguas Calientes, Shintuya) and another six locations in the Amarakaeri Communal Reserve. The area is humid and hot: during the wet season, the average daily temperature is 24.78°C, the average humidity is 90.58% and the rainfall is 3098 mm ; during the dry season the average temperature is 23.74°C, the average humidity is 84.89% and the average monthly rainfall is 1557 mm (climate information from Whitworth et al. 2016b ). Ameerega shihuemoy is most commonly found in low disturbance forest. During the dry season it is frequently found amongst boulders along or near forest streams ( Fig. 8 ). During the rainy season it moves away from the streams into the forest interior. The activity patterns appear to be distinctly crepuscular, being most active in the early morning (05:30 to 09:00) or evening (16:00 to 18:00) when males call vigorously. Males typically call from exposed positions of rocks, leaf litter, or woody debris. Calling has been heard throughout the year. Crevices or holes made of boulders or roots are used as refuges. During the night, individuals rest on low vegetation between 0.1 to 0.5 m above the ground. Reproduction takes place near both permanent and seasonal streams. On 25 June 2013 and 10 July 2013 two clutches of eggs were found inside of small shelters next to a stream. These clutches contained 22 and 25 eggs respectively, and were guarded by males. On 30 July 2013 , one uncollected male was observed transporting ten tadpoles along a slow moving stream. We detected free-living A. shihuemoy tadpoles and metamorphs co-occurring with metamorphs of A. macero in the same streams; with shallow, slow moving, clear water and bottoms of sand and dead leaves. Results of the GLM analysis in both forest types (CCR and SLR) showed that six environmental variables were the most important habitat features to explain the species presence near streams (frog presence ~ canopy cover + leaf litter cover + number of potential refuges + presence of large rocks + presence of a still body of water + stream flow; Table 4 , Appendix IV). The best supported model explained 89.05% of the variation in the data. When we ran the same analysis just in streams where the frogs were found (SLR forest), the GLM analysis presents two top models with ∆AICc <2 ( Table 5 , Appendix V). The best supported model explained 83.41% of the variation in the data. Model averaging was carried out in these two models. The relative importance for leaf litter cover, number of potential refuges, presence of large rocks, presence of a still body of water and stream flow were 1.0, signalling that they were important predictors. Canopy cover had some support (0.69), suggesting lower importance when comparing just within the SLR forest, and was therefore excluded from the top-preferred, and more parsimonious model (Appendix V). This top model suggests that a greater number of potential refuges (often created by the presence of a high quantity of large rocks), a great amount of leaf litter cover, the presence of still bodies of water, and a low stream flow (likely to benefit breeding strategy) are the most important predictors of the presence of A. shihuemoy near streams. Conservation status . Applying IUCN Red List criteria (IUCN Red List of Threatened Species, 2 0 0 1 Criteria & Categories (version 3.1.); http://www.iucnredlist.org/info/categories_criteria2001, accessed 20 February 2016 ), which indicates that if a species occurs in less than ten threat-defined locations and the extent of occurrence is less than 20,000 km 2, it should be classified as Vulnerable or Endangered. Ameerega shihuemoy is known from nine localities distributed in the buffer zone of the Manu Biosphere Reserve and the Amarakaeri Communal Reserve ( Fig. 9 ), with an estimated extent of occurrence of ca. 1,124 km 2, as such, we suggest that this new species might be classified as Vulnerable. However, due to the lack of intensive sampling effort in the Amarakaeri Communal Reserve, which may host a greater number of locations, we propose that A. shihuemoy should likely be categorized as Near Threatened (NT). Model K AICc ∆AICc ωi frog presence~ A + B + C + D + E + F 6 39.015 0.000 0.395 frog presence~ A + B + C + D + E + F + G 7 41.169 2.154 0.135 frog presence~ A + B + C + D + E + F + H 7 41.210 2.194 0.132 frog presence~ B + C + D + E + F 5 41.567 2.552 0.110 frog presence~ B + C + D + E + F + G 6 43.303 4.288 0.046 frog presence~ A + B + C + D + E + F + G + H 8 43.388 4.372 0.044 frog presence~ B + C + D + E + F + H 6 43.593 4.578 0.040 frog presence~ A + B + C + D + F 5 45.232 6.217 0.018 frog presence~ B + C + D + E + F + G + H 7 45.414 6.399 0.016 frog presence~ B + C + D + F 4 46.134 7.118 0.011 frog presence~1 1 224.474 185.459 0.000 Predictor variables are canopy cover (A), leaf litter cover (B), number of potential refuges (C), percentage of large rocks (D), presence of a still body of water (E), stream flow (F), leaf litter depth (G) and Wood debris percent (H). Model K AICc ∆AICc ωi frog presence~ A + B + C + D + E + F frog presence~ B + C + D + E + F frog presence~ A + B + C + D + E + F + H 6 5 7 38.109 39.667 40.385 0.000 1.558 2.276 0.284 0.130 0.091 frog presence~ A + B + C + D + E + F + G frog presence~ A + B + C + D + F frog presence~ B + C + D + F 7 5 4 40.400 41.082 41.728 2.292 2.973 3.619 0.090 0.064 0.046 frog presence~ B + C + D + E + F + G frog presence~ B + C + D + E + F + H frog presence~ A + B + C + D + E + F + G + H 6 6 8 41.733 41.914 42.712 3.625 3.805 4.603 0.046 0.042 0.028 frog presence~ B + D + E + F frog presence~1 4 1 42.793 164.154 4.684 126.045 0.027 0.000 Predictor variables are canopy cover (A), leaf litter cover (B), number of potential refuges (C), percentage of large rocks (D), presence of a still body of water (E), stream flow (F), leaf litter depth (G) and Wood debris percent (H).

Measurements (in mm) of the holotype . The female holotype ( Fig. 1 ) has SVL, 25.7; TL, 12.3; GBW, 8.2; HW, 7.9; HL, 8.0; ED, 5.2; IOD, 3.0; TSCN, 2.0; NED, 2.6; IND, 2.9; EL, 3.3; HDT, 2.0; MTD, 0.8; HaL, 7.0; WTFD, 0.7; WTF, 0.5; WTTD, 0.8; WTT, 0.4; WFTD, 0.9; WFT, 0.4. Description of the type series. External morphology: A small to medium-size Ameerega , with adult males attaining approximately 19.2–21.8 mm SVL and adult females approximately 21.5–25.7 mm SVL (measurements and proportions summarized in Table 1 ). Dorsal skin of head, body, shank, thigh, and hind limbs coarsely and conspicuously granular; skin smooth or nearly smooth on forelimbs, and smooth on sides of head, body and on ventral surfaces. Head slightly wider between jaw articulations than between outer edges of upper eyelids; head usually narrower than body or about as wide. Head width between jaws 29–33% of SVL. Snout sloping, bluntly pointed or rounded in profile, truncate to rounded (usually) or bluntly pointed in dorsal and ventral view. Nares situated near tip of snout, directed slightly posterolaterally; nares visible from front and from below but not from above. Canthus rostralis short, protruding; loreal region nearly vertical, slightly concave to flat. Interorbital distance wider than upper eyelid. Eye shorter than snout length; distance from center of naris to eye 60–87% of eye length. Tympanum 35–65% of eye size. Hand ( Fig. 1 F) moderately large, its length 23–28% of SVL and 74–90% of head width between angles of jaws. Relative lengths of adpressed fingers IV> I> II> III; fingers I, II, and IV approximately similar in size when adpressed, with overlapping discs; adpressed first finger varies from slightly shorter to slightly longer than second. Finger discs weakly to moderately expanded; third finger disc 1.2–1.8 times wider than distal end of adjacent phalanx. A large outer metacarpal (= palmar) tubercle on the base of the palm and a smaller inner metacarpal tubercle on the base of first finger, these being relatively small, with rounded surfaces. One subarticular tubercle on fingers I & II, and two subarticular tubercles on fingers III & IV; subarticular tubercles well developed and prominently raised, although distal one on finger III and both in IV sometimes weaker; supernumerary tubercles, finger keels, and outer metacarpal fold absent; tibia length 47–54% of SVL; relative lengths of adpressed toes, IV> III> V> II> I; first toe usually reaching base of subarticular tubercle of second toe; basal webbing on toes II–IV; toe fringes absent; outer metatarsal fold absent; a large inner metatarsal fold located on the distal half of tarsus and extended near the inner metatarsal tubercle ( Fig. 1 G); ventrolateral side of tarsus relatively smooth, not especially rugose or tubercular; toes with moderately expanded discs, wider than finger discs ( Fig. 1 G); one to three moderately raised subarticular tubercles (one each on toes I & II, two each on III & V, and three on IV); two large metatarsal tubercles with low, rounded surfaces; inner metatarsal tubercle slightly larger than outer metatarsal tubercle; supernumerary tubercles absent. Color pattern in life. (Based on living specimens and photographs; Figs. 2 , 3 ). The noticeably granular body is blackish with bronze shades middorsally, turning black and blue dorsolaterally. The flanks are mostly black with metallic bluish green, with a distinct bright orange, coral, or cream stripe extending obliquely from groin to above the eye, and forward along the canthus rostralis to join around the snout. A pale whitish-bronze labial stripe commences between naris and eye, and extends posteriorly under the eye and tympanum to the base of the upper arm. A bright orange to coral spot is present on the hip and upper thigh; there is no calf spot and no pale markings in axilla or groin. The limbs are bronze-brown with shades of green and black. The ventral surfaces are overall blue with variable black reticulation, while in some individuals the ventral surfaces are overall black with blue reticulation. Color in preservative ( Fig. 1 ). The bright orange and coral colors fade to pale grey; the blue ventral surfaces fade to grey in a dark reticulum of variable distinctiveness. MHNC 15448 MUSA 3181 MUSA 3178 MHNC 10525 MHNC 12988 MHNC 10526 MHNC 10406 MHNC 10407 MUSA 3180 MUSA 3182 MHNC 10408 MHNC 10536 MHNC 10542 SEX Female Female Female Female Female Female Female Female Female Female Male Male Male SVL 25.7 25.2 25.1 24.4 24.0 23.5 23.2 23.1 22.8 21.5 21.8 21.6 19.2 TL 12.3 12.9 11.4 11.8 12.2 11.2 11.5 11.2 11.6 11.7 11.0 10.9 9.0 HaL 7.0 6.8 6.3 6.3 6.5 6.0 6.2 6.0 6.1 5.5 6.1 5.8 4.5 HL 8.0 7.2 7.9 7.8 7.5 7.7 8.2 8.3 6.7 6.9 6.8 7.9 7.4 HW 7.9 7.6 7.8 7.4 7.6 6.8 7.3 7.4 6.8 7.2 6.8 6.7 6.0 GBW 8.2 8.4 7.9 8.5 7.2 7.6 8.3 7.4 6.8 6.7 7.9 7.6 7.6 IOD 3.0 3.1 3.0 2.3 2.7 2.1 2.3 2.3 2.8 2.3 2.1 2.3 2.1 ED 5.2 5.2 5.1 5.0 5.1 5.1 4.7 5.1 4.9 4.8 4.5 4.7 4.3 HDT 2.0 1.9 1.5 1.4 1.5 1.3 1.5 1.4 1.5 1.1 1.4 1.1 1.0 EL 3.3 3.0 3.3 3.1 3.1 3.1 2.9 2.9 3.1 3.0 2.7 2.7 2.9 TSCN 2.0 1.6 1.8 1.3 1.6 1.5 1.2 1.5 1.8 1.8 1.0 1.1 1.1 NED 2.6 2.6 2.3 2.2 2.1 2.0 1.9 2.1 2.1 1.8 2.3 1.8 2.0 IND 2.9 2.7 2.9 2.9 2.9 2.5 2.3 2.8 2.6 2.8 2.7 2.6 2.2 MTD 0.8 0.8 0.9 0.7 0.9 0.7 0.7 0.5 0.7 0.6 0.8 0.6 0.7 W3FD 0.7 0.8 0.6 0.7 0.8 0.7 0.6 0.7 0.7 0.5 0.6 0.5 0.7 W3F 0.5 0.5 0.3 0.4 0.4 0.4 0.3 0.4 0.4 0.3 0.4 0.4 0.4 W4TD 0.9 0.9 0.8 0.7 0.8 0.7 0.5 0.7 0.7 0.5 0.6 0.7 0.6 W4T 0.4 0.4 0.4 0.3 0.3 0.4 0.3 0.4 0.3 0.3 0.4 0.5 0.4 W3TD 0.8 0.8 0.7 0.7 0.7 0.5 0.7 0.7 0.7 0.6 0.4 0.6 0.6 W3T 0.4 0.5 0.4 0.4 0.4 0.4 0.3 0.4 0.3 0.3 0.3 0.4 0.3 Vocalization. Recordings of three A. shihuemoy males were taken at the Manu Learning Centre reserve. The advertisement call of this new species can be characterized as a series of chirp-like, pulsed notes. Notes are repeated at a rate of 0.8–1.0 notes per second (0.9 ± 0.1), duration of individual notes range from 84–109 ms (98.4 ± 6.8 ms), spaced 969–1196 ms apart (1042.5 ± 186.6 ms), with eight pulses (8.2 ± 0.8) per note. Dominant frequency ranges from 4478.9–4909.6 Hz (4672.72 ± 251.0 Hz) and is not frequency-modulated. Calling activity happens most frequently in the early mornings between 05:00 to 09:00 and late afternoon between 16:00 to 18:00. We also recorded a second call in A. shihuemoy consisting of three notes in quick succession (within 174.9 ± 16.6 ms of each other), repeated once every four to five seconds. The three notes in this call have different duration; the first note (87.2 ± 2.5 ms) typically has a longer duration than the second (69.4 ± 2.6 ms) and the third note (72.8 ± 2.8 ms). This three-note call seems to function as an aggressive or territorial call. We compared the call of the following species with Ameerega shihuemoy : two recordings of A. simulans (74 advertisement calls) by J. M. Padial from Marcapata, Quispicanchis Province , Department Cusco , Peru , were obtained from Fonoteca Zoologica ( Museo Nacional de Ciencias Naturales 2016); two recordings of A. boliviana (45 advertisement calls) by J. Bosch & I. De la Riva from Correo-Apolo , La Paz , Bolivia ; one recording of A. yungicola (30 advertisement calls) by M. Pacheco-Suarez from Caranavi, Yungas, Bolivia obtained from “Guía fotográfica de los anfibios de la region de las yungas Bolivia ” ( Pacheco-Suarez 2015 ); two recordings of A. hahneli (46 advertisement calls) by J. Serrano-Rojas from Shintuya , Madre de Dios , Peru ; two recordings of A. picta (63 advertisement calls) by J. M. Padial from Madidi National Park, Northern Bolivia and from Paractito , Cochabamba Department , Central West Bolivia deposited at Fonoteca Zoologica ( Museo Nacional de Ciencias Naturales 2016); two recordings of A. macero (55 advertisement calls) by J. Serrano-Rojas from Manu Learning Centre , Madre de Dios , Peru . The advertisement call of A. shihuemoy is easily distinguished ( Fig. 4 ) from the morphologically similar Ameerega species of the Ameerega picta group (for measurable call parameters and intra- and interspecific variation see Table 2 ). Parameters A. shihuemoy A. simulans A. picta A. hahneli Mean SD Mean SD Mean SD Mean SD Calls/N° of individuals 80/3 - 74/2 - 63/2 - 46/2 - Notes/min 52.8 5.4 79.2 3.6 130.2 6.0 511.8 4.2 Notes/s 0.9 0.1 1.3 0.1 2.2 0.1 8.5 0.1 Duration of the note (ms) 98.4 6.8 104.9 10.2 45.8 2.4 13.4 2.8 Break between calls (ms) 1042.5 186.6 691.4 162.7 429.9 44.6 107.4 12.6 Fundamental frequency (Hz) 4237.0 281.9 4060.9 74.6 3770.7 76.7 2516.8 83.7 Dominant frequency (Hz) 4672.7 251.0 4460.3 157.7 4044.2 94.7 4550.0 49.1 continued. Parameters A. boliviana A. yungicola A. macero Mean SD Mean SD Mean SD Calls/N° of individuals 45/ 2 - 30 - 55 /2 - Notes/min 72.0 4.2 313.2 0.0 519.4 2.2 Notes/s 1.2 0.1 5.2 0.0 8.7 0.0 Duration of the note (ms) 80.9 8.6 47.8 4.7 37.6 1.3 Break between calls (ms) 783.2 88.6 148.1 7.3 76.1 2.7 Fundamental frequency (Hz) 3416.1 68.2 3475.7 43.5 3353.7 38.1 Dominant frequency (Hz) 3846.0 46.3 3703.7 0.0 3617.6 0.0 The note duration of the new species (84–109 ms) is longer than A. hahneli (11–18 ms), A. macero (36–40 ms), A. picta (41–52 ms) and A. yungicola (40–57 ms), but similar to A. boliviana (75–94 ms) and A. simulans (91–118 ms). The space between calls in A. shihuemoy is longer than in the other six species. The number of notes per second in the advertisement call of A. shihuemoy is lower than the other six species. The call of the new species has a higher fundamental frequency than all other species but similar to A. simulans , and a higher dominant frequency than all other species but similar to A. hahneli and A. simulans , except the latter species has pronounced upward frequency-modulated notes by about 950 Hz ( Fig. 5 ). It is important to take into consideration that this comparison was performed on recordings from three individuals of A. shihuemoy from a single location. Tadpoles. Ontogenetic variation of 11 characteristics measured for 16 tadpoles in stages 24–46 ( Gosner 1960 ) are summarized in Table 3 . The tadpole of A. shihuemoy sp. nov. belongs to the exotrophic ecomorphological guild, benthic type as defined by Altig & Johnson (1989) . This description is based on a tadpole (MHNC lot 12987) at developmental Stage 25 ( Gosner 1960; Figs. 3 A, C ) and measuring a total length 17.2 mm ; body length 6.2 mm ; maximum width 4.1 mm ; depth 3.0 mm; internarial distance 0.6 mm ; eye to nares distance 0.5 mm ; eye diameter 0.6 mm ; interorbital distance 1.0 mm, oral disc width 1.2 mm ; tail length 11.0 mm. The body is globular, compressed in lateral view, and ovoid in dorsal view. The snout is rounded in dorsal view ( Figs. 6 A and C). The mouth is located anteroventrally and is surrounded by a small oral disc ( Figs. 6 B and D). Papillae are laterally emarginated, simple and conical. Marginal papillae absent on anterior labium, present in one complete row on posterior labium. Anterior jaw sheath has a medial indentation with reduced serration, posterior jaw sheath Vshaped and has serration throughout. Lateral processes long, extending well past lower jaw. Labial Tooth Row Formula (LTRF) is 2(2)/3(1). A-1 complete, A-2 with medial gap, same width as A-1. P-1, P-2, and P-3 complete; P-1 and P-2 equal width, P-3 shorter. Nares are oval, small, without projections and inflexions, and are located dorsolaterally. Eyes are small and oriented dorsolaterally. The spiracle is single, sinistral, and located just before mid-body. Fins are concave and the posterior end is rounded, reaching their maximum height at the last third of the tail; the dorsal fin does not extend onto the body. The maximum tail height is reached at about the end length and is as high as body height. In preservative, the body is dark grey, the belly is translucent with intestines slightly visible; caudal musculature creamy white with small, irregular grey flecks and slightly translucent fins. In life, the body is dark brown with black spots, the belly is transparent but slightly pigmented posteriorly, intestines well visible through skin, caudal musculature poorly pigmented and transparent tail fin with melanophores in small and irregular clusters along the tail. Gosner stages In addition, we described observations in the field following Gosner (1960) . We found two clutches of eggs in small rocky cavities; they were monitored every two days in order to take notes about the development of eggs. The first set was found on 25 June 2013 , with 22 eggs at stage 13; a stage in which the neural plate develops on the dorsal surface, and eggs were covered by a transparent orange mucilage. After nine days, eggs become tadpoles, where they reach stage 17 (tail bud). The same embryos were abandoned by the father after two days on 0 5 July 2013 . The embryos were getting dirty and dry, and inside we could observe the larvae of an insect, which could likely prey upon the embryos. Character 24 (N=1) 25 (N=3) 26 (N=2) 27 (N=1) 40 (N=2) 41 (N=2) 42 (N=1) 43 (N=1) 44 (N=1) 45 (N=1) 46 (N=1) TL 17.8 16.9 ± 1.9 17 ± 2.8 18.3 22.6 ± 0.2 23.1 ± 0.6 16.6 14.5 12.1 10.5 10.4 BL BW 6.4 3.9 6 ± 0.9 3.6 ± 0.5 6.5 ± 0.8 3.9 ± 0.4 9.5 4.7 9.4 ± 0.1 5.3 ± 0 9.4 ± 0.4 6.8 ± 1.1 8.5 4.3 9.2 4.6 11.9 3.5 9.5 4.8 10.4 4.6 BH TAL TMW 2.2 11.4 1.1 2.6 ± 0.4 10.9 ± 1.1 1.5 ± 0.2 3 ± 0.2 10.5 ± 2 1.3 ± 0.1 3.5 11.8 2.0 3.8 ± 0.4 13.2 ± 0.1 2.2 ± 0.2 4.5 ± 0.1 13.7 ± 0.1 2.8 ± 0.1 3.2 8.1 2.2 4.2 5.3 2.7 3.3 0.2 1.4 2.4 1.0 1.9 2.4 _ _ TMH END IND 1.7 1.0 0.6 1.6 ± 0.2 0.6 ± 0.3 0.6 ± 0.1 2.1 ± 0.2 0.6 ± 0.2 0.9 ± 0.2 2.0 1.1 1.0 3.1 ± 0.2 0.9 ± 0.1 0.9 ± 0.2 3.5 ± 0.1 1 ± 0.3 1.2 ± 0.2 2.3 0.6 1.1 2.7 1.6 1.1 1.1 1.0 0.9 1.7 0.8 0.9 _ 1.8 0.8 ED IOD OD 0.6 0.8 1.1 0.6 ± 0.2 0.9 ± 0.2 1.1 ± 0.4 0.7 ± 0.1 1.2 ± 0 1.2 ± 0 1.0 1.7 1.2 1.3 ± 0 1.7 ± 0.4 2.15 ± 0.1 1.3 ± 0.1 2 ± 0.1 2.2 ± 0 1.4 2.0 2.3 1.3 2.5 1.8 1.5 2.1 2.6 1.4 2.3 3.2 1.5 2.4 3.9 The second set was found on 10 July 2013 , containing 25 eggs in stage 13 covered by a transparent orange mucilage. After ten days, eggs were observed at stage 19 (heart beat), which is represented mainly by the development of the gills and tail. Seven days later tadpoles were at stage 21 (cornea transparent), in which the tail becomes increasingly transparent. Finally, three days later ( 30 July 2013 ) only five tadpoles were found at stage 22 (tail fin circulation); we assumed that the father was carrying the rest of tadpoles. One year later we found two other tadpoles at stage 26 (limb bud), on 11 June 2014 . After four days, these reached stage 38 (toe development XIII). Another group of four tadpoles were found at stage 40 (cloacal tail piece XV–XVII), which changed to stage 42 (nose development XXI) after four days. Two more tadpoles were found at stage 42 (nose development XXI) on 15 June 2014 ; the stage 46 (metamorphosis complete) was reached after three days. In summary, we suggest that the development between stages 13–22 takes twenty days, stages 26–38, four days, and stages 40–46, seven days. However, the time of development of stages 22–26, and stages 36–40 remains unknown. Phylogenetics. Our Maximum Likelihood (ML) tree was generally congruent with previous molecular phylogenies based on mitochondrial DNA ( Twomey & Brown 2008 , Brown & Twomey 2009 ) and supported the distinctiveness of the Ameerega shihuemoy from other closely related taxa. The new species was most closely related to Ameerega macero , A. altamazonica , A. rubriventris ( Lötters et al. 1997 ) , and two undescribed species— Ameerega sp. from Porto Walter, Acre , Brazil , and Ameerega sp. from Ivochote, Cusco , Peru ( Fig. 7 ). In general, the topology we recovered is similar to the ones obtained by Twomey & Brown (2008) and Brown & Twomey (2009) , although there were some differences. These differences were likely a result of using only one mitochondrial gene (our study) versus three mitochondrial genes. Nevertheless, in addition to determining the relationship between the new species and other closely related taxa, our analysis inferred three main cladesbassleri , hahneli and petersi —identified in these previous studies ( Twomey & Brown 2008 , Brown & Twomey 2009 ). Our analysis also inferred several species pairs that were inferred to be closely related according to the previous analyses (e.g., A. picta and A. yungicola ; A. rubriventris and A. altamazonica ).