New tribal placement and review of Parapucaya Prell and Pucaya Ohaus (Coleoptera, Scarabaeidae, Dynastinae)

Abstract The dynastine scarab genera Parapucaya Prell and Pucaya Ohaus have been historically classified in Pentodontini; however, that tribal classification is not supported under the current tribal circumscriptions. A discussion justifying the transfer of the genera Parapucaya and Pucaya from Pentodontini into Cyclocephalini is presented. This research is based on morphological observations (mandible shape and wing characters among others) and molecular data (genes 28S, COI, and 16S/ND1). A review of both genera is included, providing descriptions, diagnoses, distribution data, illustrations, and keys to species. A revised key to the world genera of Cyclocephalini is also included.


Introduction
Dynastinae is classified in the scarab beetle family Scarabaeidae, a large coleopteran family that comprises about 30,000 species (Ratcliffe and Cave 2015). Though Scarabaeidae is well-studied, almost 200 new species are described each year (Ratcliffe and Cave 2015). Some adults of Scarabaeidae stand out due to their relatively large size, bright colors, elaborate ornamentation, unique life histories, and many interesting adaptations (Jameson 1998). These exaggerated features are common in the subfamily Dynastinae, which includes about 1,500 species distributed worldwide (Ratcliffe and Cave 2017). More dynastine species are found in the Neotropics than in any other biogeographic realm (Ratcliffe and Cave 2015). In the Neotropics, six of the eight recognized dynastine tribes are represented: Cyclocephalini, Pentodontini, Oryctini, Phileurini, Agaocephalini, and Dynastini. The Neotropical genera Parapucaya and Pucaya have long been classified in the tribe Pentodontini based on morphological characters, but some recent authors have questioned their tribal placement (Clark 2011, López-García et al. 2015. In this study, we address the classification of Parapucaya and Pucaya within Pentodontini and redefine the tribe Cyclocephalini.

Cyclocephalini
Cyclocephalini is the second most species-rich tribe of Dynastinae after Pentodontini, and it contains 14 genera and over 500 species and subspecies (Smith 2006, Moore et al. 2015, 2018b, Ratcliffe and Cave 2017. Historically, the tribe Cyclocephalini was characterized by the absence of characters found in other dynastines. These characters included: 1) lack of horns, tubercles, carinae, or foveae on the head and prothorax; 2) absence of a stridulatory area (pars stridens) on the propygidium; 3) simple mandibles that lack dentition distal to the molar region; 4) metatibial apex truncate and without produced teeth or a crenulated margin; and 5) metatarsus with basal joint simple and not triangular (Ratcliffe and Cave 2017). The sexual dimorphism found in cyclocephalines is not as pronounced as it is in the horned dynastines. However, most cyclocephaline species display sexual dimorphism of the protarsus (enlarged in males; simple in females) and elytral epipleural margin (simple in males; expanded and modified in females of some species). Moore (2012) hypothesized that during mating, there was an interaction between the enlarged male protarsal claw and the female epipleural expansion, making it easier for the male to clasp the female during copulation and for mate guarding. Moreover, as in all dynastines, the apex of the last abdominal sternite is emarginate in males and entire or rounded in females (Figs 1, 2).
Cyclocephalini, while relatively morphologically uniform, is not well defined, and monophyly of the tribe still needs to be evaluated (Ratcliffe and Cave 2017). In that endeavor, some generic-level taxa have been removed from the tribe while others have been transferred into Cyclocephalini. Coscinocephalus Prell was transferred from Cyclocephalini to Pentodontini and is considered to be most similar to Orizabus Fairmaire (Morón and Ratcliffe 1996). The bizarre, monotypic genus Acrobolbia Ohaus was transferred from Rutelinae to Cyclocephalini by Jameson et al. (2002), and those authors compared the genus to Ancognatha Erichson. Peltonotus Burmeister, with 25 species, was transferred from Rutelinae to Dynastinae (Jameson 1998) and specifically to Cyclocephalini (Jameson and Jakl 2010). Additionally, the monophyly of several cyclocephaline genera is in doubt. Ratcliffe (2003) stressed that further research is needed on the genera Cyclocephala Dejean, Mimeoma Casey, Aspidolea Bates, and Ancognatha to ascertain if they should be maintained as valid genera or some should be folded into others. Moore et al. (2015) evaluated the monophyly of Mimeoma and its relationship with Cyclocephala by using a combined molecular and morphological analysis. These data showed that the five species of Mimeoma clustered within an apical clade of other Cyclocephala species, rendering Cyclocephala paraphyletic. As a result, Mimeoma was synonymized with Cyclocephala.

Pentodontini
Pentodontini is the largest tribe of dynastines, comprising about 100 genera and over 550 species distributed worldwide Morón 1997, Ratcliffe andCave 2017). Adult pentodontines are distinguished by: 1) the presence of tubercles, a carina, or a fovea on the head and pronotum; 2) broad mandibles with or without teeth on the scissorial region; 3) propygidium with or without a pars stridens; 4) lateral margin of the protibia usually tridentate; 5) apex of the metatibia usually truncate and margined with short, spine-like setae; 6) protarsus occasionally enlarged in males Morón 1997, Ratcliffe andCave 2017).
Dimorphism between males and females is slight in most species (Ratcliffe and Morón 1997), although males sometimes have larger protarsi and tubercles on the head and pronotum (López-García et al. 2015), and the pronotal fovea is more pronounced. Pentodontines, along with all dynastines, display sexual dimorphism of the last abdominal sternite, which is emarginate in males and entire or rounded in females (Figs 1, 2). López-García et al. (2015) reported that for over 100 years, there was no consensus whether Pentodontini should be treated as a family, subfamily, or tribe. Historical workers prioritized different criteria: Mulsant (1842) considered the categories Pentodonaires and Oryctésaires as separate groups; followed by Bates (1888) who designated Pentodontinae as a subfamily Morón 1997, López-García et al. 2015); Casey (1915) established Pentodontini as a tribe and later Leng (1920), Arrow (1937), Blackwelder (1944, and Arnett (1973) did not recognize any of these former designations and included pentodontine genera within Oryctini (Ratcliffe and Morón 1997). Endrődi (1969) re-established the tribes Pentodontini and Oryctini as they are currently used, but he considered that transitional characters blurred the distinction between the tribes. Consequently, the monophyly of Pentodontini is in doubt (Ratcliffe 2003, Gasca-Álvarez et al. 2008, López-García et al. 2015, Sanabria et al. 2012.

Morphological methods
Morphological descriptions and temporal and distributional data were based on the study of 425 specimens from three sources: (1) field collecting expeditions by the authors and colleagues; (2) data recorded from the literature; and (3)  This study was developed as part of the broader project "The Dynastine Scarab Beetles of Ecuador". For this reason, we provide only generalized, province-level distribution data for Pucaya and Parapucaya species in Ecuador. More detailed distribution data for these genera will be released as part of that forthcoming monograph. Collecting methods utilized were: 1) light traps using mercury vapor and ultraviolet bulbs; 2) foliage gleaning; 3) excavating rotting logs and stumps; and 4) manual collecting around public lights. Ecuadorian collecting, mobilization, and export permits were obtained with the support of QCAZ in Quito, Ecuador. The species descriptions encompass the range of variation observed in the specimens at hand. They were based on the following characteristics (from Ratcliffe and Cave 2017): 1) length from apex of the clypeus to the apex of the elytra; 2) width across elytral humeri; 3) coloration and markings; 4) interocular width (number of transverse eye diameters across the frons between the eyes); 5) form and sculpturing of the head, pronotum, elytra, and pygidium; 6) form of the prosternal process; and 7) form of the parameres. Punctures were considered simple unless otherwise noted. Minute punctures were generally not visible with 12.5× magnification but were easily seen with 50× magnification. Small punctures were clearly visible with 12.5× magnification and can be seen with the naked eye. Large punctures are easily seen without magnification. Sparse punctures were characterized by greater than 5 puncture diameters between them. Punctures moderate in density had 3-5 puncture diameters between them. Dense punctures had only 2 or fewer puncture diameters between them.

DNA extraction, PCR, and data-mining
Previous studies by Gunter et al. (2016) and Ahrens et al. (2011Ahrens et al. ( , 2014 generated DNA sequence data that served as a phylogenetic scaffold for testing the classification of Pucaya and Parapucaya within Pentodontini (Tab. 1). GenBank was datamined for 28S, CO1, and 16S/ND1 sequences from diverse tribal-level exemplars for higher Scarabaeidae (Tab. 1). Among Dynastinae, there were tribal-level exemplars with at least partial data for all three gene regions from six of the eight commonly recognized tribes (minus Hexodontini and Agaocephalini). 16S and 28S data were generated from exemplar specimens of Pucaya pulchra Arrow and Parapucaya amazonica Prell to incorporate into this phylogenetic framework. Based on shared morphological characters with Pucaya species, Cyclocephala freyi Endrődi exemplars were also targeted for DNA extraction and PCR.

Alignments and phylogenetic analyses
Based on the results of Gunter et al. (2016), a species of Isonychus Mannerheim (Scarabaeidae: Melolonthinae: Macrodactylini) was used as an outgroup for all phylogenetic analyses. Isonychus sp. was recovered as the most early-diverging member of a melolonthine clade sister to the clade containing all Cetoniinae + Rutelinae + Dynastinae exemplars (Gunter et al. 2016), making this taxon a suitable outgroup for examining relationships among these subfamilies and placing Pucaya and Parapucaya at the tribal level. Sequences were aligned using ClustalW (Larkin et al. 2007), with default settings, as implemented in MEGA7 (Kumar et al. 2016). The resulting concatenated sequence alignment contained 3,537 bp positions (1530 bp 16S; 550 bp ND1; 805 bp CO1; 652 bp 28S). Maximum likelihood analyses of this matrix were conducted in W-IQ-TREE (Trifinopoulos et al. 2016). The matrix was partitioned by gene (16S, 28S, and ND1) and codon position (CO1). The best-fit model of sequence evolution for each partition (GTR+F+I+G4 for 16S; TPM3u+F+G4 for ND1 and CO1 third position; TIM2e+I+G4 for 28S; HKY+F+I+G4 for CO1 first position; SYM+I+G4 for CO1 second position) was selected by ModelFinder (Kalyaanamoorthy et al. 2017), as implemented in W-IQ-TREE, using the Bayesian information criterion. Bootstrap support values for the most likely tree were calculated using 10,000 ultrafast bootstrap replicates (Hoang et al. 2017). Bayesian phylogenetic analyses were run in MrBayes 3.2.2 (Ronquist and Huelsenbeck 2003). Models of sequence evolution for the MrBayes analysis were determined with PartitionFinder 2.1.1 (GTR+I+G for 16S, ND1, 28S, CO1 second position, and CO1 third position; HKY+I+G for CO1 first position) (Lanfear et al. 2016).
Bayesian analyses comprised four independent runs, each with four chains (one cold and three heated). Partitions had their parameters unlinked and allowed to vary independently. Flat priors were used. Chains were run for 1 million generations, with trees sampled every 1,000 generations. Convergence was evaluated by examining the standard deviation of split frequencies among runs and by plotting the log-likelihood values from each run using Tracer 1.6 (Rambaut and Drummond 2013). Tracer diagnostics indicated that runs converged within 10,000 generations, and trees sampled during this period were discarded as burn-in before obtaining clade posterior probabilities. Parsimony tree searches were performed in MPBoot (Hoang et al. 2018). Heuristic searches were conducted using default parsimony ratchet search options in MPBoot. Bootstrap analyses were performed using the same ratchet search options and included 10,000 bootstrap replicates.

Morphology
Morphological observations show that Parapucaya shares characters with genera in Cyclocephalini, most notably with some Cyclocephala species. For example, the two Parapucaya species share characters with C. almitana Dechambre, C. macrophylla Erichson, C. melanocephala (Fabricius), and C. pseudomelanocephala Dupuis. These characters include: 1) frontoclypeal suture complete; 2) clypeus weakly emarginate with lateral and apical margins reflexed; 3) clypeal apex broadly truncate; 4) the generally exposed and slender mandibles that lack lateral teeth; 5) mandibular apex acuminate and curved upward; 6) protibia strongly tridentate with the basal tooth removed from other two teeth; 7) protarsus in males enlarged (the larger claw strongly curved and incised at apex), while females have a simple protarsus; 8) inner portion of the apical margin of the 5 th protarsomeres in males eroded, allowing the enlarged protarsal claw to be further articulated; 9) metatarsi reduced, shorter than metatibia, more evident in females (character shared with C. melanocephala and C. almitana); 10) prosternal process moderately long, columnar, with its apex densely setose, flattened, and with a large, raised, round "button" covering half of the apex; 11) hindwing vein RA, proximal to apical hinge, with 2 rows of pegs extending distally nearly to margin of apical hinge; and 12) anterior edge of hindwing distal to apical hinge lacking setae and with a produced, membranous border (Figs 3, 4).
Like Parapucaya, Pucaya species share many characters with some Cyclocephala species (e.g., C. freyi). Pucaya also shares the character of a medially incomplete frontoclypeal suture with Ancognatha species. In Pucaya individuals, the frontoclypeal suture is visible from the lateral margins along the external side of the frontal horn, where it becomes obsolete medially. Pucaya and some Ancognatha species display weakly developed "armature" of the head and thorax. For example, Ancognatha castanea Erichson has tubercle-like swellings on the frontoclypeal region of the head. Ancognatha jamesoni Murray and A. horrida Endrődi show enlargement of the pronotum in males.
Other shared characters with other cyclocephalines include; 1) clypeus with lateral and apical margins reflexed; 2) clypeal apex broadly truncate, subquadrate; 4) maxillary galea with four teeth on inner margin (shared with C. freyi (Figs 5, 60), 5) slender mandibles that lack lateral teeth; 6) protibia strongly tridentate with the basal tooth removed from other two teeth; 7) protarsus in males enlarged (the larger claw strongly curved and incised at apex), while females have a simple protarsus; 8) inner portion of the apical margin of the 5 th protarsomeres in males eroded, allowing the enlarged protarsal claw to be further articulated; 9) prosternal process short to moderately long, columnar, with its apex densely setose, flattened, and with a large, raised, round "button" covering half of the apex; 11) hindwing vein RA, proximal to apical hinge, with 2 rows of pegs extending distally nearly to margin of apical hinge; and 12) anterior edge of hindwing distal to apical hinge lacking setae and with a produced, membranous border.

Molecular phylogenetic analyses
W-IQ-TREE analyses found the most likely tree with a log likelihood score of -42928.5840. MPBoot heuristic tree searches recovered most parsimonious trees of score 9992 steps. Bayesian posterior probabilities and parsimony bootstrap support values for nodes are reported on the maximum likelihood bootstrap consensus tree topology (Fig. 7). Analyses conducted on the concatenated dataset recovered 27 strongly supported internal nodes (>75 BS and >0.95 PP) from all three tree search strategies. All three analyses strongly supported the monophyly of Cetoniinae and Dynastinae (Fig. 7). Like the analyses of Gunter et al. (2016), the subfamily Rutelinae was recovered as paraphyletic. Parapucaya amazonica, P. pulchra, and C. freyi were recovered together as a clade (94 ML BS, 0.97 PP, 73 Parsimony BS) sister to the other three Cyclocephala exemplars. Together, these six exemplars form a strongly supported cyclocephaline clade (99 ML BS, 1.0 PP, 79 Parsimony BS) within the broader Dynastinae clade (99 ML BS, 1.0 PP, 91 Parsimony BS) (Fig. 7). The remaining 14 Pentodontini species included here did not form a monophyletic group. Six pentodontine species fell out in a clade that includes Cryptodus sp. (Dynastinae: Phileurini) (96 ML BS, 1.0 PP). Eight pentodontine species were recovered in a clade (98 ML BS, 1.0 PP, 86 Parsimony BS) that also included Oryctes nasicornis (Linnaeus) (Dynastinae: Oryctini).

Discussion
Parapucaya and Pucaya were placed in Pentodontini by previous authors, and this triballevel classification has been maintained since Endrődi's (1985) revision of world Dynastinae. Parapucaya and Pucaya species were placed in Pentodontini because of their armature, such as the minute tubercles of the pronotum in Parapucaya species and the cephalic horns and tubercles of Pucaya species. These characters violated the tribal circumscription of Cyclocephalini. However, these two genera also complicate the traditional circumscription of Pentodontini. For example, Parapucaya and Pucaya have slender mandibles, and males and females can be easily distinguished by external characters.
Based on the morphological observations outlined in the previous section, we think that Parapucaya species are most similar to the C. melanocephala section of Cyclocephala. Additionally, we think that Pucaya species are most similar to C. freyi based on the shared form of the four-toothed galea present in all these species (Figs 5,6). The following characters also support the hypothesis that Pucaya and Parapucaya are cyclocephalines: clypeus with lateral and apical margins reflexed; the clypeal apex broadly truncate shared with several Cyclocephala species; mandibles lacking lateral teeth; protibia strongly tridentate with the basal tooth removed from other two apical teeth; protarsus in males enlarged (the larger claw strongly curved and incised at apex), while females have a simple protarsus; and the inner portion of the apical margin of the 5 th protarsomeres in males eroded, allowing the enlarged protarsal claw to be further articulated.
Study of the hindwings also showed that Pucaya and Parapucaya share the same character states: hindwing vein RA, proximal to the apical hinge, with two rows of pegs extending distally nearly to margin of apical hinge and the anterior edge of hindwing distal to apical hinge lacking setae and with a produced, membranous border. This exact combination of hindwing characters is also found in the cyclocephaline genera Arriguttia Martínez, Aspidolea, Augoderia Burmeister, most Cyclocephala (except black species formerly placed in Mononidia Casey or Surutoides Endrődi), and former Mimeoma species (Moore et al. 2018a). The genera Acrobolbia, Ancognatha, and Ruteloryctes also share the membranous border on the leading edge of RA3 but lack the double row of pegs on RA (Moore et al. 2018a). No other tribe of Dynastinae shares the character of a membranous border on RA3 (MRM, unpublished data). This hindwing character is a putative synapomorphy uniting these cyclocephaline genera plus Pucaya and Parapucaya.
Additionally, the molecular phylogenetic analyses presented here also support revised placement of Pucaya and Parapucaya in Cyclocephalini. Our analyses recovered a monophyletic Dynastinae with strong statistical support (Fig. 7). These analyses also recovered a strongly supported clade that included four Cyclocephala exemplars plus P. castanea and P. amazonica (Fig. 7). We think the weight of evidence supports the hypothesis that Pucaya and Parapucaya are part of the cyclocephaline lineage of Dynastinae. Based on morphological observations, we also think that Pucaya and Parapucaya are most likely to be closely related to sections of Cyclocephala. Thus, we formally move the genera Pucaya and Parapucaya, as a revised tribal placement, from Pentodontini into Cyclocephalini.  Historically, Cyclocephalini has been defined by the lack of characters present in other tribes, such as the lack of horns or tubercles, foveae, or carinae. However, this was an inconsistent concept as Ancognatha species with weakly developed cephalic and thoracic armature, (e.g., tubercles, enlarged pronotum, and enlarged mandibles) were already classified in Cyclocephalini. This work categorically indicates that Cyclocephalini includes individuals with armature. This is a potentially fascinating re-circumscription of the tribe, as the role of cephalic and thoracic armature is completely unknown for Pucaya, Parapucaya, and Cyclocephalini more broadly.

Review of Parapucaya Prell and Pucaya Ohaus
We present a revised key to the New World Cyclocephalini genera. We include a redescription of the species of Parapucaya and Pucaya, diagnosis, distribution data, and available natural history information. We include keys to species of both genera.

Key to the world genera of adult Cyclocephalini
(Modified from Jameson et al. 2002) Males: Apex of last abdominal sternite emarginate (Fig. 1). Protarsomeres 4-5 and/ or anterior claw enlarged in all genera except Stenocrates and Erioscelis. Females: Apex of last abdominal sternite entire, evenly parabolic (Fig. 2). Protarsomeres 4-5 and anterior claw always simple, not enlarged. Clypeus with sides slightly wider than base before abruptly narrowing to acuminate apex (Fig. 13). Males with antennal club almost twice as long as antennomeres 1-7 (Fig. 13) Lateral margins of clypeus near base raised into a subacute crest, evident in posterodorsal view (Fig. 25). Clypeus thickened along the frontoclypeal suture. Frontoclypeal disc concave (Fig. 25) Lateral margins of clypeus near base flat or faintly raised into a round crest, evident in posterodorsal view (Fig. 26). Clypeus flat or weakly thickened along the frontoclypeal suture. Frontoclypeal disc convex or concave (Fig. 26). Specimens without double tubercles or faint declivity near anterior margin of pronotum ...9 9 Clypeus trapezoidal or subtrapezoidal, with marginal or apical bead ( Fig. 27-28 Notes. Parapucaya contains two Neotropical species. The genus is distinct from other Cyclocephalini because of the presence of a strongly impressed frontoclypeal suture with the clypeus raised along the suture; lateral margin of clypeus near base raised into a subacute crest, evident in posterodorsal view (Fig. 25); and the presence of double tubercles or declivity near the anterior margin of the pronotum. It is necessary to look closely at the anterior margin of the pronotum to see the two small tubercles that help to characterize this genus, which can occasionally be difficult in some specimens where these tubercles are nearly absent, especially in P. amazonica. The color and general appearance of specimens of Parapucaya make them appear similar to C. melanocephala and other related species. Adults of Parapucaya have been collected at lights at night. Species of this genus are found distributed in tropical lowlands, such as coastal and Amazonian rainforests, but also in areas with temperate climate, such as cloud forests. Based on label data of Ecuadorian individuals, specimens have been found in pastures. Nothing is known about the immature stages of Parapucaya species.

Key to the species of Parapucaya
Males with protarsomeres enlarged, protarsus with one claw simple and one enlarged. Females with protarsomeres slender, protarsus with both claws simple.
Diagnosis. Parapucaya amazonica is invariably mistaken for species of Cyclocephala because of its similar appearance. The subapical declivity of the pronotum (or two tubercles in well-developed specimens), in combination with the raised basal margins of the clypeus and the raised clypeal surface along the frontoclypeal suture, will distinguish this genus from Cyclocephala species.
Parapucaya amazonica and P. nodicollis can be separated from each other by the shape of the mentum (concave from disc to apex in P. amazonica, evenly convex in P. nodicollis), the pronotal tubercles (subtle in P. amazonica, conspicuous in P. nodicollis), the presence or absence of pygidial setae (base and lateral angles of pygidium setose in P. amazonica, glabrous in P. nodicollis); size (in general, P. amazonica is larger and stouter than P. nodicollis, although some individuals overlap); and their parameres (Fig. 32).
Natural history. In Ecuador, P. amazonica occurs at elevations ranging from sea level to 2,450 m in the coastal, Andean, and Amazon regions. Based on label data, adults can be collected throughout the year but in higher numbers in February and December. Nothing is known of the immature stages of this species.
Diagnosis. Parapucaya nodicollis is usually mistaken for species of Cyclocephala because of its similar appearance. The two small tubercles on the pronotum, in combination with the raised basal margins of the clypeus and the raised clypeal surface along the frontoclypeal suture, will distinguish members of this genus from Cyclocephala species.
Parapucaya nodicollis and P. amazonica can be separated from each other by the shape of the mentum (evenly convex in P. nodicollis, concave from disc to apex in P. amazonica); the pronotal tubercles (conspicuous in P. nodicollis, subtle in P. amazonica); the presence or absence of pygidial setae (glabrous in P. nodicollis, present across the base of the pygidium in P. amazonica); size (in general, P. nodicollis is smaller and thinner than P. amazonica, although some individuals overlap); and their parameres (Fig. 34).
Natural history. In Ecuador, it occurs at elevations from 300 to 1,800 m on both sides of the Andes. Based on label data, adults can be collected in Ecuador throughout the year and in higher numbers in February, June to July, and in November. Nothing is known of the immature stages of this species.
Pucaya Ohaus, 1910Figs 4, 6, 35-40 Pucaya Ohaus, 1910 Notes. The genus Pucaya contains two species, P. castanea Ohaus and P. pulchra Arrow. López-García et al. (2015) compared type specimens and synonymized P. punctata  Endrődi with P. pulchra based on similarities in body length, pronotal and elytral punctation, and the fact that the description of P. punctata was based on the color and punctation of a single female in a species where color pattern and punctation are variable.
Pucaya is distinguished from other cyclocephalines by its broadly truncate clypeus that conceals the mandibles; a small horn or tubercle near each eye (horns not as developed in Ecuadorian specimens as in Panamanian specimens); parameres with round, minute spinules (bumps) on the apical half; and a characteristic binodose pronotum.

Key to the species of Pucaya
Diagnosis. Pucaya castanea can be distinguished from P. pulchra by its elytral punctation. In P. castanea, the entire elytral surface has sparse, minute punctures, while in P. pulchra the elytral surface is striate-punctate from the base to 2/3 the length of the elytra. The punctures are dense, moderate in size, and ocellate, but on the apical third of the elytra the punctures are sparse and minute. The form of the parameres (Fig. 38) also separates both species.
Natural history. In Ecuador, P. castanea occurs at elevations ranging from sea level to 2,550 m in the coastal, Andean, and Amazon regions. Based on label data, adults can be collected throughout the year but in higher numbers from February to May and from November to December. Nothing is known of the immature stages of this species. Arrow, 1911 Figs 6, 35, 39-40 Pucaya pulchra Arrow, 1911: 167 (original combination).

Pucaya pulchra
Redescription. Length 20.4-23.7 mm; width 9.8-11.2 mm. Color of head black or piceous. Pronotum completely black or black with brown, elongate markings on   margins, with or without brown spots on base of disc. Elytra entirely black or black with brown margins or brown with black markings on the suture, humerus and behind scutellum; markings can be short near elytral base or extend to umbone area. Scutellum, pygidium, venter, and legs black or brown. Head: Frons sparsely punctate at base, becoming progressively rugo-punctate anteriorly; punctures moderate in size. Frontoclypeal sutural area at sides with tubercle in both sexes; tubercle smaller in females, conical in males. Clypeus with apex very broadly truncate, reflexed, surface rugose at disc, smooth to shagreened at margins. Interocular width equals 4.1-4.3 transverse eye diameters. Antenna with 10 antennomeres, club slightly longer than antennomeres 2-7. Pronotum: Surface moderately to densely punctate at base, punctures moderate in size; sparsely punctate from disc to apex, punctures minute. Broadly depressed midline, with round depressions on each side of midline: 1 on apex, 2 between mid-disc and margins; depressions shallow in females. Elytra: Surface from base to 2/3 striate-punctate; punctures dense, moderate in size, ocellate; from 2/3 to apex with sparse, minute punctures. Pygidium: Surface densely punctate, punctures moderate in size. In lateral view, males with surface evenly rounded, females with surface nearly flat. Legs: Protibia tridentate. Protarsus in male weakly enlarged, median claw large, strongly curved, cleft at apex; protarsus and claw simple in female. Venter: Prosternal process moderately long, columnar; apex densely setose, flattened, and with large, raised, round "button" covering most of apex; setae long, tawny. Parameres: Fig. 40.
Diagnosis. Pucaya pulchra can be distinguished from P. castanea by the elytral punctation . In P. pulchra, the elytral surface is striate-punctate from the base to 2/3 of the elytra; the punctures are dense, moderate in size, and ocellate, but the apical third has sparse, minute punctures. In P. castanea, the entire elytral surface has sparse, minute punctures. The form of the parameres (Fig 40.) also separates both species.
Natural history. In Ecuador, P. pulchra occurs at elevations ranging from 20 to 1,900 m in the coastal, Andean, and Amazon regions. Some specimens have been collected in pitfall traps.
Álvaro Barragán, Taryn Ghia, and Fernanda Salazar (all Museo de Zoología de la Pontificia Universidad Católica del Ecuador), and mobilization permits needed within Ecuador were provided by the directors and staff at the different Ministerio del Ambiente offices throughout Ecuador. We thank Omar Torres (Pontificia Universidad Católica del Ecuador) for his support and encouragement to pursue a molecular analysis of dynastines. He, together with Santiago Ron, collaborated with funding through their project Arca de Noé and with their personnel. We thank Claudia Terán (Pontificia Universidad Católica del Ecuador) for obtaining the DNA sequences necessary for the molecular analysis for several Dynastinae species and for her great spirit of collaboration. We thank Gabriela Castillo and Andrea Manzano (both Pontificia Universidad Católica del Ecuador) who also participated in the first phases of the molecular work.
For loans and/or on-site access to institutional specimens, we thank Brett Ratcliffe and M.J. Paulsen (University of Nebraska State Museum), Álvaro Barragán, Florencio Maza, and Fernanda Salazar (all Museo de Zoología de la Pontificia Universidad Católica del Ecuador, Quito, Ecuador); Vladimir Carvajal, David Donoso, Miguel Pinto, and Adrián Troya (all Museo de la Escuela Politécnica Nacional, Quito, Ecuador); Santiago Villamarín and Diego Inclán (Museo Ecuatoriano de Ciencias Naturales, Quito, Ecuador); Diego Marín (Colección de Invertebrados del Sur del Ecuador, Universidad Técnica Particular de Loja, Loja Ecuador); François Génier and Robert Anderson (Canadian Museum of Nature, Ottawa, Canada); Patrice Bouchard and Serge Laplante (Canadian National Collection of Insects, Ottawa, Canada); Paul Skelley (Florida State Collection of Arthropods, Gainesville, Florida, United States); Joseph Jelínek (National Museum of Natural History, Prague, Czech Republic); and Stephane Le Tirant (personal collection, Terrabonne, Québec, Canada). We are grateful to the Fundación Jocotoco and the staff at the Canandé Reserve for the opportunity to visit their reserve and for their hospitality during a collecting trip.
Special thanks to Ronald Cave (University of Florida), Estefanía Micó (Universidad de Alicante), and Brett Ratcliffe (University of Nebraska) for their valuable comments that helped improve this manuscript. Estefanía Micó provided us with the protocols for the 16S and COI genes. Brett Ratcliffe and Ronald Cave (University of Florida) are acknowledged for sharing specimen data, conducting field research, and gathering data from museums in Ecuador. M.J. Paulsen (University of Nebraska) is acknowledged for his help with the photographic equipment at the University of Nebraska State Museum. We thank Beulah Garner (The Natural History Museum), Mary Liz Jameson and Oliver Keller (both Wichita State University) for their collaboration analyzing morphological and molecular characters in the early stages of this project. Mary Liz Jameson and Brett Ratcliffe are also acknowledged for sharing their Cyclocephalini illustrations. We thank Nicole Gunter (Cleveland Museum of Natural History) for her help in preparing the sequence alignments for phylogenetic analysis. We are grateful to Gavin J. Martin (Brigham Young University) for his help with imaging fine morphological structures of Pucaya and Parapucaya, and to Brett Ratcliffe (University of Nebraska) for the Pucaya pulchra image.
Partial support for field research and collections work was provided by a grant from the National Geographic Society (NGS 9936-16) to Brett C. Ratcliffe. This work was also supported by the Secretaría de Educación Superior, Ciencia, Tecnología e Innovación (SENESCYT) under the "Arca de Noé" Initiative (PIs: S. R. Ron and O. Torres-Carvajal).