Abstract

Staphylus evemerus Godman & Salvin, 1896 is a species with a unique set of morphological characters within Carcharodini. Also, mitochondrial cytochrome oxidase subunit 1 (COI) sequences analysis demonstrated a large genetic distance with other related genera of the tribe. Therefore, this paper aims to describe a new genus for this species, which is named as Uniphylus gen. nov. Besides the morphological redescription of the male of Uniphylus evemerus (Godman & Salvin, 1896) new comb., the description of the female is provided for the first time, as well as an updated distributional map with all records known so far for this species.

Key words
Skipper; Neotropical; Costa Rica; Panama; cytochrome oxidase subunit 1

INTRODUCTION

Carcharodini is a tribe of Pyrginae (Hesperiidae, Lepidoptera) that includes approximately 300 species distributed in 35 genera, four of which are present in the Old World (Carcharodus Hübner, [1819], Spialia Swinhoe, 1912, Gomalia Moore, 1879 and Muschampia Tutt, 1906), while the others occur in the New World regions (Warren et al. 2009Warren AD, Ogawa JR & Brower AVZ. 2009. Revised classification of the family Hesperiidae (Lepidoptera: Hesperioidea) based on combined molecular and morphological data. Syst Entomol 34: 467-523., Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., Li et al. 2019Li W, Cong Q, Shen J, Zhang J, Hallwachs W, Janzen DH & Grishin NV. 2019. Genomes of skipper butterflies reveal extensive convergence of wing patterns. Proc Natl Acad Sci 116: 6232-6237., Zhang et al. 2020aZhang D, Gao F, Jakovlić I, Zou H, Zhang J, Li WX & Wang GT. 2020c. PhyloSuite: An integrated and scalable desktop platform for streamlined molecular sequence data management and evolutionary phylogenetics studies. Mol Ecol Res 20: 348-355.). Carcharodini is a sister-group of Pyrgini, with the divergence time occurring in the Eocene around 42 Mya, together with the other two tribes of Pyrginae, Achlyodini and Erynnini (Li et al. 2019Li W, Cong Q, Shen J, Zhang J, Hallwachs W, Janzen DH & Grishin NV. 2019. Genomes of skipper butterflies reveal extensive convergence of wing patterns. Proc Natl Acad Sci 116: 6232-6237.). Though the monophyly of Carcharodini is strongly recovered in molecular analysis performed in above cited studies, no morphological synapomorphy has been recognized so far. Despite this, Warren et al. (2009)Warren AD, Ogawa JR & Brower AVZ. 2009. Revised classification of the family Hesperiidae (Lepidoptera: Hesperioidea) based on combined molecular and morphological data. Syst Entomol 34: 467-523. partially characterized the males of Carcharodini to possess an asymmetrical pattern in the genitalia morphology, known as “Nisoniades-type” valva (sensu Evans 1953Evans WH. 1953. A Catalogue of the American Hesperiidae Indicating the Classification and Nomenclature Adopted in The British Museum (Natural History). Part III. (Groups E, F, G) Pyrginae. Section 2. London: British Museum, 246 p.). Most of the female genitalia of Carcharodini, as it is for many other groups of Lepidoptera, are not described. However, Siewert et al. (2016)Siewert RR, Zacca T, Mielke OHH & Casagrande MM. 2016. Taxonomic revision and cladistic analysis of the genus Sophista Plötz, 1879 (Lepidoptera: Hesperiidae) with description of a new species from north-east Brazil. Zool J Linn Soc 177: 526-540. illustrated and described the female genitalia of Sophista as asymmetrical as well. Concerning the immature stages, Carcharodini larval foodplants include a variety of dicots, although Cyclosemia herennius (Stoll, 1782) is the only New World Carcharodini known to feed on monocots (Warren et al. 2009Warren AD, Ogawa JR & Brower AVZ. 2009. Revised classification of the family Hesperiidae (Lepidoptera: Hesperioidea) based on combined molecular and morphological data. Syst Entomol 34: 467-523.).

Although skippers are historically less studied than other butterfly families, this scenario is currently changing, with numerous studies taxonomically redefining subfamilies, tribes, genera, and subgenera with the use of molecular tools, most of them including genome sequencing (Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., Li et al. 2019Li W, Cong Q, Shen J, Zhang J, Hallwachs W, Janzen DH & Grishin NV. 2019. Genomes of skipper butterflies reveal extensive convergence of wing patterns. Proc Natl Acad Sci 116: 6232-6237., Zhang et al. 2019aZhang J, Qian C, Shen J, Brockmann E & Grishin NV. 2019a. Genomes reveal drastic and recurrent phenotypic divergence in firetip skipper butterflies (Hesperiidae: Pyrrhopyginae). Proc R Soc Lond B 286: 1-6., bZhang J, Qian C, Shen J, Brockmann E & Grishin NV. 2019b. Three new subfamilies of skipper butterflies (Lepidoptera, Hesperiidae). ZooKeys 861: 91-105., 2020aZhang J, Brockmann E, Cong Q, Shen J & Grishin NV. 2020a. A genomic perspective on the taxonomy of the subtribe Carcharodina (Lepidoptera: Hesperiidae: Carcharodini). Zootaxa 4748: 182-194., b, 2022a). Moreover, some of these studies suggest that different molecular analyses can produce good hypotheses regarding generic taxonomical levels (Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., Zhang et al. 2022aZhang J, Cong Q, Shen J & Grishin NV. 2022a. Taxonomic changes suggested by the genomic analysis of Hesperiidae (Lepidoptera). Insecta Mundi 0921: 1-135.). For example, some of the new genera recently proposed for Carcharodini, such as Clytius Grishin, 2019 (type species: Pholisora clytius Godman and Salvin, [1897]), Incisus Grishin, 2019 (type species: Antigonus incisus Mabille, 1878), Perus Grishin, 2019 (type species: Pholisora cordillerae Lindsey, 1925), and Viuria Grishin, 2019 (type species: Pellicia licisca Plötz, 1882), have been recovered in four different molecular approaches: nuclear genome, z chromosome, mitochondrial genome, and COI barcode (Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56.). Also, when the morphological characters are evaluated, especially those of the male genitalia, these new taxa demonstrate good morphological diagnostic characters.

Staphylus Godman & Salvin, 1896 (type species: Helias ascalaphus Staudinger, 1876) is one of the richest genera of Carcharodini, with 47 species. A related genus is Bolla Mabille, 1903 (type species: Bolla pullata Mabille, 1903, currently a junior subjective synonym of Bolla imbras (Godman and Salvin, [1896]), comprising 29 species (Mielke 2005Mielke OHH. 2005. Catalogue of the American Hesperioidea. Volumes 1-6, Curitiba: Sociedade Brasileira de Zoologia, 1536 p., Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., Zhang et al. 2022aZhang J, Cong Q, Shen J & Grishin NV. 2022a. Taxonomic changes suggested by the genomic analysis of Hesperiidae (Lepidoptera). Insecta Mundi 0921: 1-135., b, Lemes et al. 2023LEMES JRA, SIEWERT RR, MIELKE OHH, CASAGRANDE MM & WARREN AD. 2023. Taxonomic and distributional notes on Bolla tepeca (Bell, 1942), new combination (Lepidoptera: Hesperiidae: Pyrginae). Trop Lepid Res 33: 102-110.). Together, these two genera of small brown Carcharodini represent very challenging groups, as their accurate identifications are usually possible only after dissecting and analyzing their genitalia (Lemes et al. 2021LEMES JRA, SIEWERT RR, MIELKE OHH & CASAGRANDE MM. 2021. First record of the skipper Bolla mancoi (Lindsey, 1925) (Hesperiidae) for Brazil in an Amazonian Protected Area. J Lepid Soc 75: 217-220., 2023). Another close genus is Pholisora Scudder, 1872 (type species: P. catullus (Fabricius, 1793)), currently with only four valid species (Mielke 2005Mielke OHH. 2005. Catalogue of the American Hesperioidea. Volumes 1-6, Curitiba: Sociedade Brasileira de Zoologia, 1536 p., Warren et al. 2023WARREN AD, DAVIS KJ, STANGELAND EM, PELHAM JP, WILLMOTT KR & GRISHIN NV. 2023. Illustrated Lists of American Butterflies. [14-II-2023]. Available from: http://www.butterfliesofamerica.com. (Accessed: 05 March 2023).
http://www.butterfliesofamerica.com... ), although historically Pholisora has included several species of the above cited genera (Mielke 2005Mielke OHH. 2005. Catalogue of the American Hesperioidea. Volumes 1-6, Curitiba: Sociedade Brasileira de Zoologia, 1536 p.).

When studying some Carcharodini species, it was noted that the Central American species Staphylus evemerus Godman & Salvin, 1896 is morphologically distinct from other species of Staphylus Godman & Salvin, 1896, with larger wings, forewings with inner margin concave and a unique male and female genitalia pattern. The morphological combination of characters of S. evemerus also does not fit into any of the known Carcharodini genera. To test a generic taxonomical hypothesis for S. evemerus, COI-sequences of this species and 20 other of Staphylus were obtained. A large genetic distance was observed between S. evemerus and other Staphylus, corroborating the recognition of a new genus. Therefore, the aim of this paper is to describe a new Carcharodini genus to include S. evemerus, and to provide additional taxonomical notes on this species.

The ZooBank Life Science Identifier (LSID) of this publication is: urn:lsid:zoobank.org:pub:702C0ABD-7005-45E2-B4E5-943F5696C5DB

MATERIALS AND METHODS

The material used in this study is from the following public and private collections: DZUP - Coleção Entomológica Pe. Jesus de Santiago Moure, Universidade Federal do Paraná, Curitiba, Paraná, Brazil; MGCL - McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, Florida, United States; NHMUK - The Natural History Museum, London, United Kingdom; OM - Olaf Hermann Hendrik Mielke collection, Curitiba, Paraná, Brazil. Eight specimens of Uniphylus evemerus new comb. were found in these institutions.

For the molecular analysis, two legs (meso and metathoracic from the same side) from 22 skippers of the internal group and related Carcharodini genera (Table I) were detached and sent to the Canadian Center for DNA Barcoding (CCDB) for DNA extraction, amplification, and sequencing of the mitochondrial cytochrome oxidase subunit 1 (COI) using standardized methods (Hebert et al. 2003Hebert PDN, Cywinska A, Ball SL & deWaard JR. 2003. Biological identifications through DNA barcodes. Proc R Soc Lond 270: 313-321.). The obtained sequences were included in GenBank with accession numbers available in table I. These specimens were identified through analysis of their genitalia and comparison with original descriptions and other important taxonomical studies, such as Evans (1953)Evans WH. 1953. A Catalogue of the American Hesperiidae Indicating the Classification and Nomenclature Adopted in The British Museum (Natural History). Part III. (Groups E, F, G) Pyrginae. Section 2. London: British Museum, 246 p. and Steinhauser (1989)STEINHAUSER SR. 1989. Taxonomic notes and descriptions of new taxa in the Neotropical Hesperiidae. Part I. Pyrginae. Bull Allyn Mus 127: 1-70.. COI sequences available in GenBank (NCBI 2020NCBI - National Center for Biotechnology Information. 2020. Bethesda (MD): National Library of Medicine (US), National Center for Biotechnology Information; [1988] - [Accessed in 10 Feb. 2023]. Available from: https://www.ncbi.nlm.nih.gov/.
https://www.ncbi.nlm.nih.gov/... ) of other Carcharodini species were also included in the analysis (Janzen et al. 2011Janzen DH, Hallwachs W, Burns JM, Hajibabaei M, Bertrand C & Hebert PDN. 2011. Reading the Complex Skipper Butterfly Fauna of One Tropical Place. PLoS ONE 6: e19874., Pratt et al. 2015Pratt GF, Rugman-Jones PF & Stouthamer R. 2015. Genetic Characterization of North American Sootywings (Lepidoptera: Hesperidae) Using Nuclear and Mitochondrial Loci Validates Specific Status of MacNeill’s Sootywing, Hesperopsis gracielae (MacNeill). Proc Entomol Soc Wash 117: 194-202., Lavinia et al. 2017Lavinia PD, Núñez-Bustos EO, Kopuchian C, Lijtmaer DA, García NC, Hebert PDN & Tubaro PL. 2017. Barcoding the butterflies of southern South America: Species delimitation efficacy, cryptic diversity and geographic patterns of divergence. PLoS ONE 12: e0186845., Pfeiler 2018Pfeiler E. 2018. DNA Barcoding and Taxonomic Challenges in Describing New Putative Species: Examples from Sootywing and Cloudywing Butterflies (Lepidoptera: Hesperiidae). Diversity 10: 1-14., Attiná et al. 2021Attiná N, Núñez-Bustos EO, Lijtmaer DA, Hebert PDN, Tubaro PL & Lavinia PD. 2021. Genetic variation in neotropical butterflies is associated with sampling scale, species distributions, and historical forest dynamics. Mol Ecol Resour 21: 2333-2349., Earl et al. 2021Earl C ET AL. 2021. Spatial phylogenetics of butterflies in relation to environmental drivers and angiosperm diversity across North America. IScience 24: 1-18., Santos et al. 2022Santos B ET AL. 2022. Enhancing DNA barcode referencelibraries by harvesting terrestrial arthropods at the National Museum ofNatural History. Arpha Preprints: 1-21.). The chosen species for the analysis were selected based on the recent phylogenetic hypothesis for Carcharodini (Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56. and Zhang et al. 2022aZhang J, Cong Q, Shen J & Grishin NV. 2022a. Taxonomic changes suggested by the genomic analysis of Hesperiidae (Lepidoptera). Insecta Mundi 0921: 1-135.). For this analysis, a total of 42 specimens from 39 species belonging to ten Carcharodini genera were used (Table I).

The molecular alignment for COI sequences was performed using the software Muscle in MEGA7 (Kumar et al. 2016Kumar S, Stecher G & Tamura K. 2016. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Mol Biol Evol 33: 1870-1874.). The pairwise genetic distances (%) was also calculated in MEGA7, and the frequency distribution of genetic divergence was plotted using pairwise values. The optimal model of nucleotide evolution was selected by ModelFinder (Kalyaanamoorthy et al. 2017Kalyaanamoorthy S, Minh BQ, Wong TKF, von Haeseler A & Jermiin LS. 2017. ModelFinder: fast model selection for accurate phylogenetic estimates. Nat Methods 14: 587-589.) using the AICc criterion, resulting in GTR+F+I+G4. Maximum likelihood analysis was conducted using the software IQ-Tree (Nguyen et al. 2015Nguyen LT, Schmidt HA, von Haeseler A & Minh BQ. 2015. IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies. Mol Biol Evol 32: 268-274.) implemented in the PhyloSuit (Zhang et al. 2020c). For the Maximum Likelihood analysis, the ultra-fast bootstrap approximation method - UFBoot (5,000 bootstrap repetitions) was used (Minh et al. 2013Minh BQ, Nguyen MAT & von Haeseler A. 2013. Ultrafast Approximation for Phylogenetic Bootstrap. Mol Biol Evol 30: 1188-1195.), together with the SH-aLRT test (1,000 repetitions) (Guindon et al. 2010Guindon S, Dufayard J-F, Lefort V, Anisimova M, Hordijk W & Gascuel O. 2010. New Algorithms and Methods to Estimate Maximum-Likelihood Phylogenies: Assessing the Performance of PhyML 3.0. Syst Biol 59: 307-321.). The tree obtained was visualized and initially edited in the software FigTree v.1.4.3 (Rambaut 2006Rambaut A. 2006-2009. FigTree v1.4.3. Institute of Evolutionary Biology, University of Edinburgh. Available from: http://tree.bio.ed.ac.uk/software/figtree/.
http://tree.bio.ed.ac.uk/software/figtre... -2009), and later in the software Inkscape 1.0.1 (Harrington et al. 2004Harrington B, Gould T, Hurst N & MenTaLgu Y. 2004-2005. Inkscape. Available from: http://www.inkscape.org/.
http://www.inkscape.org/... -2005). The tree was rooted at Viola minor (Hayward, 1933Hayward KJ. 1933. Lepidopteros Argentinos. Familia Hesperiidae III. Rev Soc Entomol Arg 5: 219-275.). Nodes were considered supported when both SH-aLRT ≥ 80 and UFBoot ≥ 95.

Abdomens of the studied specimens were detached and soaked in a heated 10% potassium hydroxide solution (KOH) for 5-10 minutes, and posteriorly dissected for the study of the genitalia. Dissected genitalia were stored in vials with glycerin, labeled accordingly, and photographed using a photo stacking process associated with a Leica® Application Suite Version 4.12.0 stereomicroscope. In the “examined material” section, the symbol “*” indicates that the genitalia of the specimens were dissected and analyzed. The general terminology follows Evans (1953)Evans WH. 1953. A Catalogue of the American Hesperiidae Indicating the Classification and Nomenclature Adopted in The British Museum (Natural History). Part III. (Groups E, F, G) Pyrginae. Section 2. London: British Museum, 246 p., and the genitalia morphology follows Carneiro et al. (2013)Carneiro E, Mielke OHH & Casagrande MM. 2013. Thorax and abdomen morphology of some Neotropical Hesperiidae (Lepidoptera). Insecta Mundi 327: 1-47.. The following abbreviations were used throughout the paper: DW (dorsal wing), FW (forewing), DFW (dorsal forewing), VFW (ventral forewing), HW (hindwing), DHW (dorsal hindwing), and VHW (ventral hindwing).

A taxonomical catalogue including all publications that mention the species is provided, following Mielke (2005)Mielke OHH. 2005. Catalogue of the American Hesperioidea. Volumes 1-6, Curitiba: Sociedade Brasileira de Zoologia, 1536 p.. The full references cited in the catalogue are available in Lamas (2023)Lamas G. 2023. Bibliography of Butterflies. An Annotated Bibliography of the Neotropical Butterflies and Skippers (Lepidoptera: Papilionoidea). Available from: http://butterfliesofamerica.com/L/Biblio.htm. (Accessed: 11 March 2023).
http://butterfliesofamerica.com/L/Biblio... . A distributional map was produced with SimpleMappr (Shorthouse 2010Shorthouse DP. 2010. SimpleMappr, an online tool to produce publication-quality point maps. Available from: http://www.simplemappr.net. (Accessed: 28 September 2022).
http://www.simplemappr.net... ) from the label data of the specimens analyzed and literature records, consulting the geographic coordinates of the specific locality on Google Earth. In the cases where no specific locality was given, coordinates from the central region of the state were used.

RESULTS

Molecular data

A Maximum Likelihood analysis using the mitochondrial gene COI supported the monophyly of Uniphylus gen. nov. (ultrafast bootstrap support [ubs] = 100; SH-aLRT support = 100) (Fig. 1). The Uniphylus gen. nov. clade is sister to Staphylus, and these two forms a clade with Pholisora (Fig. 1)

Interspecific genetic distance ranged from 9% to 14% (Supplementary Material - Table SI, Fig. 2); the genetic distance between Uniphylus gen. nov. and Staphylus was 10.9%, and between Uniphylus gen. nov. and Pholisora was 11%. Based on the sequences of the mitochondrial gene COI, in conjunct with the morphological differences, these results support the description of new genus to include Staphylus evemerus.

Figure 1
Relationship among Uniphylus evemerus (Godman & Salvin, 1896) new comb. and other genera of Carcharodini based on DNA barcode sequences obtained by Maximum likelihood inferred with IQTREE. Support values were considered sufficient when SH-aLRT ≥ 80 and UFBoot ≥ 95 at the same time (“*” represents sufficient support values).

Figure 2
Frequency distribution of pairwise individual genetic distances within (gray) and between (black) the species of Pholisora, Staphylus and Uniphylus gen. nov.

Taxonomy

Uniphylus Lemes, Siewert, Mielke, Casagrande & Warren gen. nov.

ZooBank Life Science Identifier (LSID) - urn:lsid:zoobank.org:act:9E54C0B9-AFD7-4221-BA2F-C86DA45FF90D

(Figs. 1-8)

Type species (Fig. 3)

Figure 3
Lectotype of Staphylus evemerus Godman & Salvin, 1896 deposited in the NHMUK in dorsal (a) and ventral (b) views, and its labels (c). Illustrations of the dorsal view of the adult and the male genitalia (d) of Staphylus evemerus available in Godman & Salvin (1896, volume B, Tab. 89). Scale bar = 1 cm.

Staphylus evemerus Godman & Salvin, 1896.

Diagnosis

Uniphylus gen. nov. is readily distinguished from other genera of Carcharodini due to the following combination of characters (Figs. 3, 4, 5a-b, 6a-d, 7a-c, 8a-h): costal fold present on the male FW; FW inner margin concave; ampulla very narrow, with upper margin concave; harpe about five times longer than wide, as a finger-like structure, with base surpassing the upper margin of ampulla, as a narrow conical process curved outwardly; tergum VIII with an anterior apophysis; lamella antevaginalis as a very sclerotized plate, quadrate in ventral view, with an excavation in the middle of the distal margin, and other larger excavation in the proximal margin in the region where presumably is the ostium bursae; lamella postvaginalis developed as narrow plate, with distal margin and proximal margin nearly straight, bearing microtrichia.

Male description (Figs. 3a-b, d, 4a-b, 5, 6, 8g-h). Head: Brown with yellow scales dorsally; labial palpus pale-yellow ventrally, except the third segment which is brown. Antenna brown, ventrally with small yellow dots at the joints at the side. Nudum with 11 segments. Thorax: Brown with scarce yellow scales dorsally; brown with white hair-like scales ventrally. Legs brown with yellow scales and white hairs. FW length: Ranging between 1.3 cm and 1.5 cm (n = 4). FW shape: Outer margin rounded. Inner margin concave. DFW: Brown. Two transversal bands with paler pattern in the discal and postdiscal areas. Presence of costal fold. Absence of white dots in the subapical area. Presence of some sparse pale scales. Fringes brown. VFW: Brown with marginal and postdiscal areas clearer. Fringe brown. HW length: Ranging between 0.9 cm and 1.1 cm (n = 4). HW shape: Outer margin slightly undulated. DHW: Brown with sparse pale scales. Two transversal bands with paler pattern in the discal and postdiscal areas. Fringe as at DFW. VHW: Brown. Presence of sparse pale scales, predominantly on the internal margin. Fringe brown. Abdomen: Brown with scarce yellow scales dorsally and ventrally. Genitalia: Tegumen slightly longer than wide, with one central constricted region, proximal portion rounded at the proximal margin and larger than the distal margin. Ventral arms of the tegumen narrow and fused with dorsal projection of saccus, assuming that the boundaries between these structures are located at the angle between them. Saccus triangular, short, rounded proximally. Uncus conical-shaped, enlarged at the base, and ventrally with a large concavity that becomes a longitudinal opening towards the narrower apex, bearing some short hair-like scales dorsally. Gnathos developed as two short, sclerotized plates bearing fine microtrichia, fused ventrally by a thinner region bearing microtrichia. Valva longer than wide; sacculus slightly longer than wide, rectangle-shaped, as a fold into the valva; harpe about five times longer than wide, except at the base where it surpasses the dorsal margin of ampulla as a narrow conical process curved outwardly; the distal part of the harpe as a finger-like structure slightly curved upwardly, with upper margin subtly indented; ampulla very narrow, with upper margin concave; costa very thin following the general shape of the valve. Sacculus, harpe, and ampulla bear thin setae throughout their extensions, internally and externally. Aedeagus cylindrical, short, about half the length of valva; without spines and vesica without cornuti. Fultura superior absent. Fultura inferior developed, thin, crescent moon-shaped in anterior view.

Figure 4
Uniphylus evemerus (Godman & Salvin, 1896) new comb. in dorsal and ventral views: a-b. Male (Costa Rica, San José, MGCL 1043687). c-d. Female (Panama, Chiriqui, Boquete, San Ramon, OM 46.830). Scale bar = 1 cm.

Figure 5
Head in dorsal (a) and ventral (b) views of a male of Uniphylus evemerus (Godman & Salvin, 1896) new comb. (Panama, Chiriqui, Alto Lino, DZ 45.615).

Figure 6
Male genitalia of Uniphylus evemerus (Godman & Salvin, 1896) new comb. in lateral (a), internal (b), dorsal (c) and ventral (d) views (Panama, Chiriqui, Boquete, San Ramon, OM 46.936).

Figure 7
Female genitalia Uniphylus evemerus (Godman & Salvin, 1896) new comb. Sterigma in ventral (a), and lateral views (b), and general ventral view (c) (Panama, Chiriqui, Boquete, San Ramon, OM 46.830).

Figure 8
Type species of some Carcharodini genera in dorsal views and male genitalia in internal view. (a-b) Bolla imbras (Godman & Salvin, 1896), (a) El Salvador, Santa Tecla, 19.VIII.1972 (MGCL); (b) Guatemala, Retalhuleu Province, NN Nuevo San Carlos, 22.XII.2002, Ulrich Reber leg., MGCL 1112131 (MGCL). (c-d) Pholisora catullus (Fabricius, 1793), (c) United States, Missouri, Jackson County, Blue Springs, 12.VIII.1967, Richard Heitzman leg., DZ 58.624 (DZUP); (d) United States, New Mexico, Carlsbad, 3.VIII.1976, no colector, DZ 58.625 (DZUP). (e-f) Incisus incisus (Mabille, 1878Mabille P. 1878. Descriptions de lépidoptères nouveaux du groupe des hespérides. Pet Nouv Ent 2: 197-198.), (e) Brazil, São Paulo, Campos do Jordão, 8-12.II.1982, Mielke & Casagrande leg., DZ 46.504 (DZUP); (f) Brazil, Santa Catarina, Joinville, 28.X.1967, Mielke & Miers leg., DZ 46.353 (DZUP). (g-h) Staphylus ascalaphus (Staudinger, 1876), (g) Guatemala, Zacapa, La Unión, 3.X.1981, Welling leg., DZ 45.176 (DZUP); (h) Mexico, Chiapas, San Jerónimo, 17.VII.1978, Welling leg., OM 44.713 (OM).

Female description (Figs. 4c-d, 7). FW length. 1.5 cm (n = 1). HW length. 1.1 cm (n = 1). Genitalia. Tergum VIII quadrate with anterior apophysis; papilla analis somewhat quadrate, covered by setae and posterior apophysis slightly curved, longer than the length of papilla analis. Lamella antevaginalis as a very sclerotized plate, quadrate in ventral view, with an excavation in the middle of the distal margin, and another larger excavation in the proximal margin in the region where presumably is the ostium bursae. Presence of a slender membranous area separating the lamella antevaginalis from the lamella postvaginalis. Lamella postvaginalis as a narrow plate, with the distal margin and proximal margin nearly straight, bearing microtrichia. Ductus bursae membranous, about three times longer than corpus bursae. Corpus bursae rounded, membranous, about ¼ the length of ductus bursae.

Comments

The wing venation of Uniphylus evemerus new comb. is very similar to that of Staphylus hayhurstii (W. H. Edwards, 1870), S. ceos (W. H. Edwards, 1882) and Pholisora catullus (Fabricius, 1793), (Lindsey 1921,Fig. 13j, k, l). The concave inner margin of the FW of Uniphylus evemerus new comb. is similar with I. incisus (Mabille, 1878) (Fig. 8e). Despite that, the male genitalia are completely different between these two species (Fig. 8f). While males of Incisus species have a broader valva, U. evemerus new comb. has a narrow valva.

The valva of the male genitalia is probably the most remarkable difference between U. evemerus new comb., and the other genera. While U. evemerus new comb. has a harpe developed as a finger-like structure, providing a unique aspect to the genitalia, in the other genera the valva is rectangular or ovoid (Fig. 8b, d, f, h). Regarding to the female genitalia, U. evemerus is the only Carcharodini species known to possess a developed anterior apophysis in the tergum VIII (Fig. 7b).

Etymology

The name Uniphylus is derived from two words: unicus, from the Latin for “only”, an allusion to the unique morphology found in the type species of the genus, especially on the male genitalia morphology; and phylus from Staphylus, where the type species was included thus far. It is a noun in the genitive singular. The gender is masculine.

Uniphylus evemerus (Godman & Salvin, 1896), new combination

(Figs. 1-8)

Staphylus evemerus Godman & Salvin, 1896. Biol. Centr.-Amer., Lep.-Rhop. 2, p. 436; 3, pl. 89, figs 31 (male d), 32 (male gen.); specimens, Caché, Rio Sucio, Irazu, Costa Rica, Rogers leg.; collection Godman & Salvin.- Draudt 1923, in Seitz. Gross-Schmett. Erde 5, p. 906, pl. 176g (d).- Evans 1953Evans WH. 1953. A Catalogue of the American Hesperiidae Indicating the Classification and Nomenclature Adopted in The British Museum (Natural History). Part III. (Groups E, F, G) Pyrginae. Section 2. London: British Museum, 246 p.. Cat. Amer. Hesp. 3, p. 96, pl. 38 (male gen.).- Bridges 1983. Lep. Hesp. 1, p. 41; 2, p. 34.- DeVries, 1983, in Janzen. Costa Rican Nat. Hist., p. 676.- Bridges, 1988. Cat. Hesp. 1, p. 65; 2, p. 57.- Bridges, 1994. Cat. Fam.-Group, Gen.-Group, Sp. Group Nam. Hesp. (Lep.) World 8, p. 78; 9, p. 64.- O. Mielke 2004. Hesperioidea, p. 55, in Lamas (Ed.). Checklist: Part 4A, Hesperioidea-Papilionoidea, in Heppner (Ed.). Atlas Neotrop. Lep. 5A; syn.: evermerus.- O. Mielke, 2005. Cat. Amer. Hesperioidea 3, p. 725; syn.: evermerus.- Anderson, 2007. Bull. Allyn Mus. 146: 8.- Janzen et al. 2011Janzen DH, Hallwachs W, Burns JM, Hajibabaei M, Bertrand C & Hebert PDN. 2011. Reading the Complex Skipper Butterfly Fauna of One Tropical Place. PLoS ONE 6: e19874.. PLoS ONE 6(8) (e19874): Suppl. Fig. S1, Table SI, Table SII.- Garwood & Lehman 2013. Butt. C. Amer. 3, Hesp., p. 108, figs (d).- Fernández-Triana et al. 2014Fernández-Triana JL ET AL. 2014. Review of Apanteles sensu stricto (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservación Guanacaste, northwestern Costa Rica, with keys to all described species from Mesoamerica. ZooKeys 383: 1-565.. Zookeys 383: 55, 141; parasitism.- Garwood et al. 2021 (online). Marip. de Colombia 1, p. 258.

Bolla evemerus; Mabille, 1903, in Wytsman. Gen. Ins. 17, p. 72.

Pholisora evermerus [sic]; Shepard, 1935. Lep. Cat. 69, p. 345.

(no genus) evemerus; Beattie, 1976. Rhop. Direct., p. 142.

Taxonomic history. Godman & Salvin (1896)Godman FD & Salvin O. 1896. Biologia Centrali-Americana. Insecta. Lepidoptera-Rhopalocera. Volume 2. London: Taylor & Francis, 1269 p. described Staphylus evemerus. The species was transferred by Mabille (1903)Mabille P. 1903. Lepidoptera Rhopalocera Fam. Hesperidae [sic]. In: Wystman PAG. Genera Insectorum 17: [ii] + 210 p. to Bolla Mabille, 1903, and after Shepard (1935)Shepard HH. 1935. Hesperiidae: Subfamilia Pyrginae. Lep Cat 69: 273-560. transferred it to Pholisora Scudder, 1872Scudder SH. 1872. A systematic revision of some of the American butterflies, with brief notes on those known to occur in Essex County, Massachusetts. Annu Rep Trust Peabody Acad Sci 4: 24-83.. Evans (1953)Evans WH. 1953. A Catalogue of the American Hesperiidae Indicating the Classification and Nomenclature Adopted in The British Museum (Natural History). Part III. (Groups E, F, G) Pyrginae. Section 2. London: British Museum, 246 p. returned the species to Staphylus, and other authors followed this decision, mentioning the species in taxonomic comments, faunistic studies, and catalogs.

Type. Staphylus evemerus Godman & Salvin, 1896 was described based on an unspecified number of specimens from Caché, Rio Sucio, Irazu (Costa Rica), collected by H. Rogers, and deposited in the Godman & Salvin’s collection. A male specimen was found in the NHMUK (Fig. 3a-c) with the following labels: / Type/ Type [H 736 written on the label back]/ R. Sucio, Costa Rica. H. Rogers / ♂/ Sp. figured./ Staphylus evemerus, sp. n. Type fig.-/ B.C.A.Lep.Rhop. Staphylus evemerus, G. & S./ Godman-Salvin Coll. 1912.-23./ BMNH(E) 1669519/. To stabilize the identity of this species, this male specimen is hereby designated lectotype of Staphylus evemerus and the following labels will be attached: / Lectotypus/ Lectotypus Staphylus evemerus Godman & Salvin, 1896; Lemes, Siewert, Mielke, Casagrande & Warren des. 2022MIELKE OHH. 1980. Contribuição ao estudo faunístico de Hesperiidae americanos. V. Nota suplementar às espécies de Pyrrhopyginae e Pyrginae do Rio Grande do Sul, Brasil (Lepidoptera). Acta Biol Paran (Curitiba) 8/9: 7-17./. These labels will be sent to the curator for the collection.

Diagnosis and description

Same as the diagnosis and description of the genus.

Biology

Janzen & Hallwachs (2009)Janzen DH & Hallwachs W. 2009. Dynamic database for an inventory of the macrocaterpillar fauna, and its food plants and parasitoids, of Area de Conservacion Guanacaste (ACG), northwestern Costa Rica. Available from: http://janzen.sas.upenn.edu. (Accessed: 03 July 2022).
http://janzen.sas.upenn.edu... provide photos of a parasitized immature in the last instar (Fig. 9a-c). The larva has a length of 28 mm., a green semi-hyaline body, and black head. Uniphylus evemerus new comb. larvae are host of the gregarious wasp Apanteles duniagarciae Fernández-Triana, 2014 (Hymenoptera, Braconidae, Microgastrinae) in the Area de Conservación Guanacaste, Costa Rica (Fernández-Triana et al. 2014Fernández-Triana JL ET AL. 2014. Review of Apanteles sensu stricto (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservación Guanacaste, northwestern Costa Rica, with keys to all described species from Mesoamerica. ZooKeys 383: 1-565.). In the above-cited area, Janzen and Hallwachs have reared 86 wild-caught caterpillars, eight of them were parasitized by A. duniagarcia, apparently the unique parasitoid of the species (D.H. Janzen, personal communication).

Figure 9
Larva of the same individual of Uniphylus evemerus (Godman & Salvin, 1896) new comb. in the last instar (a, b, c). Note the presence of the parasitoid larvae of the microgastrine braconid Apanteles duniagarciae Fernández-Triana, 2014 visible through the sides of the caterpillar (see arrows in figure a). Also, note the cocoons of the parasitoid larvae that have emerged from the caterpillar (see arrows in figure c). Costa Rica, Guanacaste Province, Area de Conservacion Guanacaste, 27.VI.1997, Ruth Franco leg., voucher 97-SRNP-1457, Area de Conservacion Guanacaste Collection. Photos by Ruth Franco.

Host plants

Pleuropetalum sprucei (Hook.fil.) Standl. (Amaranthaceae) (Janzen & Hallwachs 2009Janzen DH & Hallwachs W. 2009. Dynamic database for an inventory of the macrocaterpillar fauna, and its food plants and parasitoids, of Area de Conservacion Guanacaste (ACG), northwestern Costa Rica. Available from: http://janzen.sas.upenn.edu. (Accessed: 03 July 2022).
http://janzen.sas.upenn.edu... ).

Distribution (Fig. 10)

Uniphylus evemerus (Godman & Salvin, 1896) new comb. is recorded from the following localities in Central America: Costa Rica – Alajuela and San José; and Panama – Chiriqui.

Figure 10
Distributional map of Uniphylus evemerus (Godman & Salvin, 1896) new comb. The symbol “*” indicates literature records.

Temporal distribution

Uniphylus evemerus (Godman & Salvin, 1896) new comb. has been recorded in the following months: February, June, July, and December.

Etymology

Godman & Salvin (1896)Godman FD & Salvin O. 1896. Biologia Centrali-Americana. Insecta. Lepidoptera-Rhopalocera. Volume 2. London: Taylor & Francis, 1269 p. did not provide an etymology for the name evemerus.

Examined material

COSTA RICA: Alajuela – Socorro de la Virgen, 13.II.1988, 1♂, D. L. Lindsley leg., D. L. Lindsley colln. - MGCL Accession #2008-20, DNA Voucher JRAL-COI-11 José R. A. Lemes, MGCL 1092142 (MGCL); 14.II.1988, 1♂, D. L. Lindsley leg., D. L. Lindsley colln. - MGCL Accession #2008-20 (MGCL). Cartago - Irazu Rio Sucio, no data, ♂, H. Rogers leg., BMNH(E) 1669519, lectotype, (NHMUK). San José – No specific locality, Km 117, 26.XII.1984, 2♂, G. T. Austin leg., G. T. Austin colln. - MGCL Acc. 2004-5, MGCL 1043687, MGCL 1043693* (MGCL).

PANAMA: Chiriqui - Alto Lino, 900 m, 23.VI.1965, 1♂, Small leg., DZ 45.615* (DZUP). Boquete, San Ramon, 1500-1650 m, 20, 24.VII.1997, 1♂, Mielke & Casagrande leg., OM 46.936* (OM); Finca Lerida, Alto Quiel, 1900-2200 m, 21, 23, 25.VII.1997, 1 ♀, Mielke & Casagrande leg., OM 46.830* (OM).

DISCUSSION

Many genera of brown skippers, such as the Carcharodini, have been historically neglected in systematics studies. This is probably due to their lack of charismatic features that are found in other families of butterflies. It is not uncommon that taxonomic studies of Hesperiidae result in the description of new species and the reallocation of known species into different genera (e.g. Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., Medeiros et al. 2019Medeiros AD, Dolibaina DR, Carneiro E, Mielke OHH & Casagrande MM. 2019. Taxonomic revision of Artines Godman, 1901 (Hesperiidae: Hesperiinae, Moncini) with the description of nine new species. Zootaxa 4614: 1-049., 2020Medeiros AD, Dolibaina DR, Carneiro E, Mielke OHH & Casagrande MM. 2020. Taxonomic revision of the genus Panca Evans, 1955 (Hesperiidae: Hesperiinae: Hesperiini: Moncina) with the description of two new species. Zootaxa 4830: 503-543., Siewert et al. 2016Siewert RR, Zacca T, Mielke OHH & Casagrande MM. 2016. Taxonomic revision and cladistic analysis of the genus Sophista Plötz, 1879 (Lepidoptera: Hesperiidae) with description of a new species from north-east Brazil. Zool J Linn Soc 177: 526-540., 2020Siewert RR, Mielke OHH & Casagrande MM. 2020. Taxonomic revision of the Neotropical genus Telemiades Hübner, [1819] (Lepidoptera: Hesperiidae: Eudaminae), with descriptions of fourteen new species. Zootaxa 4721: 1-111., Zhang et al. 2022aZhang J, Cong Q, Shen J & Grishin NV. 2022a. Taxonomic changes suggested by the genomic analysis of Hesperiidae (Lepidoptera). Insecta Mundi 0921: 1-135., Lemes et al. 2023LEMES JRA, SIEWERT RR, MIELKE OHH, CASAGRANDE MM & WARREN AD. 2023. Taxonomic and distributional notes on Bolla tepeca (Bell, 1942), new combination (Lepidoptera: Hesperiidae: Pyrginae). Trop Lepid Res 33: 102-110.).

The use of genome analysis is revolutionizing the systematic studies of Hesperiidae (Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., Zhang et al. 2022aZhang J, Cong Q, Shen J & Grishin NV. 2022a. Taxonomic changes suggested by the genomic analysis of Hesperiidae (Lepidoptera). Insecta Mundi 0921: 1-135., bZHANG J, CONG Q, SHEN J, SONG L & GRISHIN NV. 2022b. Genomic DNA sequencing reveals two new North American species of Staphylus (Hesperiidae: Pyrginae: Carcharodini). The Taxonomic Report of The International Lepidoptera Survey 10: 1-14.). Ideally, sampling many loci for many taxa would be the best solution, however, this practice is still unsustainable for most research laboratories due to the expensive costs of the protocols (Talavera et al. 2021Talavera G, Lukhtanov V, Pierce NE & Vila R. 2021. DNA barcodes combined with multilocus data of representative taxa can generate reliable higher-level phylogenies. Syst Biol 71: 382-395.). Despite this, COI analyses are corroborating phylogenomic findings (Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56.), showing utility in recovering higher level relationships.

The COI-tree presented here demonstrated that U. evemerus new comb. forms a sister-group with Staphylus, and these two genera a larger clade including Pholisora (Fig. 1). However, it is important to emphasize that the inclusion of more species and more genes can change the relationships among clades. For example, Staphylus, with the new combination of S. evemerus (Godman & Salvin, 1896) in Uniphylus, includes now 47 species (Mielke 2005Mielke OHH. 2005. Catalogue of the American Hesperioidea. Volumes 1-6, Curitiba: Sociedade Brasileira de Zoologia, 1536 p., Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., Zhang et al. 2022aZhang J, Cong Q, Shen J & Grishin NV. 2022a. Taxonomic changes suggested by the genomic analysis of Hesperiidae (Lepidoptera). Insecta Mundi 0921: 1-135., b, Lemes et al. 2023LEMES JRA, SIEWERT RR, MIELKE OHH, CASAGRANDE MM & WARREN AD. 2023. Taxonomic and distributional notes on Bolla tepeca (Bell, 1942), new combination (Lepidoptera: Hesperiidae: Pyrginae). Trop Lepid Res 33: 102-110.), of which only 22 had their COI sequences analyzed in this study. Also, the inclusion of sequences of more species would likely increase the support values for the inner nodes on the COI-tree (Fig. 1).

In fact, the nuclear genome of Carcharodini suggests that Staphylus is a sister group with Hesperopsis, and Bolla is a sister group with Pholisora (Cong et al. 2019Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56.). The COI-tree of these same authors, on the other hand, suggests that Staphylus together with Hesperopsis forms a clade sister group with Bolla, which is recovered as paraphyletic, inside a larger clade including Pholisora. Therefore, the use of an integrative approach, using as much taxonomical tools as possible, is fundamental to understand the systematics of challenge groups, such as the “brown Carcharodini”. Herein, it has been demonstrated that through molecular and morphology analysis, Uniphylus gen. nov. is supported as a strong taxonomic hypothesis.

The genetic distance of COI of U. evemerus new comb. compared to other Carcharodini genera is similar with the distance among genera in the tribe, ranging from 9% to 14% (Table SI). According to Cong et al. (2019)Cong Q, Zhang J, Shen J & Grishin NV. 2019. Fifty new genera of Hesperiidae (Lepidoptera). Insecta Mundi 731: 1-56., the genetic distance between clades of Hesperiidae genera is typically more than 10% in the COI.

Uniphylus evemerus (Godman & Salvin, 1896) new comb. has a unique combination of morphological characters that separates it from other genera. Godman & Salvin (1896)Godman FD & Salvin O. 1896. Biologia Centrali-Americana. Insecta. Lepidoptera-Rhopalocera. Volume 2. London: Taylor & Francis, 1269 p. highlighted the distinctiveness of the male genitalia when compared to other Staphylus species, although these authors considered the species outwardly similar. Despite this, the strongest similarity between the wings of U. evemerus new. comb. with other Staphylus species is the brown ground color, present in most of the other genera of Carcharodini.

The concave inner margin of the FW of Uniphylus evemerus new comb. is morphologically closer to the FW of Incisus species, particularly I. incisus (Mabille, 1878) (Fig. 8e). In fact, Evans (1953, pages 95-96) had put these two species as closer species in his taxonomical key. However, the male and female genitalia pattern of U. evemerus new comb. is different than those of Incisus species (Lemes et al. in prep.). While males of Incisus species have a broader valva, U. evemerus new comb. has a narrow valva. In fact, U. evemerus new comb. has a unique genitalia pattern within Carcharodini. The very short aedeagus without spines or cornuti is another important characteristic. Although some studies recovered the male genitalia of some Carcharodini species as the “Nisoniades-type” valva (see Evans 1953Evans WH. 1953. A Catalogue of the American Hesperiidae Indicating the Classification and Nomenclature Adopted in The British Museum (Natural History). Part III. (Groups E, F, G) Pyrginae. Section 2. London: British Museum, 246 p., Warren et al. 2009Warren AD, Ogawa JR & Brower AVZ. 2009. Revised classification of the family Hesperiidae (Lepidoptera: Hesperioidea) based on combined molecular and morphological data. Syst Entomol 34: 467-523., Siewert et al. 2016Siewert RR, Zacca T, Mielke OHH & Casagrande MM. 2016. Taxonomic revision and cladistic analysis of the genus Sophista Plötz, 1879 (Lepidoptera: Hesperiidae) with description of a new species from north-east Brazil. Zool J Linn Soc 177: 526-540.), this is not the case in any species of Bolla, Pholisora, Staphylus and Incisus, as well as it is not present in U. evemerus new comb. (Fig. 8b, d, f, h, j).

For the females, the lamella postvaginalis as a narrow plate and the lamella antivaginalis as a very sclerotized quadrate plate with an excavation in the middle of the distal margin, represent very different characteristics when compared to the same genitalia structures in other Staphylus, Incisus, and Bolla species (Mielke 1975MIELKE OHH. 1975. Sobre algumas espécies de Staphylus Godman & Salvin (Lepidoptera: Hesperiidae). Acta Biol Paran (Curitiba) 4: 25-34., 1980, Steinhauser 1989STEINHAUSER SR. 1989. Taxonomic notes and descriptions of new taxa in the Neotropical Hesperiidae. Part I. Pyrginae. Bull Allyn Mus 127: 1-70., 1991STEINHAUSER SR. 1991. Six new species of Skippers from Mexico (Lepidoptera: Hesperiidae: Pyrginae and Heteropterinae). Insecta Mundi 5: 25-43., Steinhauser & Austin 1993STEINHAUSER SR & AUSTIN GT. 1993. New species of Hesperiidae from Costa Rica. Trop Lepid Res 4: 12-20., Austin 1997AUSTIN GT. 1997. Notes on Hesperiidae in northern Guatemala, with descriptions of new taxa. J Lepid Soc 51: 316-332., Austin & Warren 2002AUSTIN GT & WARREN AD. 2002. Taxonomic notes on some Neotropical skippers (Lepidoptera: Hesperiidae): Pyrrhopyginae and Pyrginae. Dugesiana 9: 15-49., Lemes et al. in prep.). The presence of an anterior apophysis of U. evemerus new comb. is until now unknown from other females of Carcharodini.

Uniphylus gen. nov. is so far a monotypic genus, but the study of other genera of Carcharodini is encouraged, as this practice may reveal more species to be included in the genus. It is possible that the low number of specimens found in the visited collections is related to the drab wings ground color and the small size of Carcharodini species, which may be less interesting among Lepidoptera collectors, as suggested by Zacca et al. (2020)ZACCA T, CASAGRANDE MM, MIELKE OHH, HUERTAS B, BARBOSA EP, FREITAS AVL & WILLMOTT KR. 2020. Description of Emeryus Zacca, Mielke & Casagrande gen. nov. (Lepidoptera: Nymphalidae) to accommodate three species formerly placed in Paryphthimoides Forster, 1964. Austral Entomol 59: 505-523.. Fortunately, this Pyrginae tribe has recently received more attention by researchers, and other studies are currently in preparation to better elucidate their taxonomic diversity (Lemes et al. in prep.).

ACKNOWLEDGMENTS

We would like to thank Thamara Zacca, Diego Rodrigo Dolibaina, Fernando Maia Silva Dias, Eduardo Carneiro dos Santos, Mônica Piovesan, Tim Shepard and the anonymous reviewer for the valuable suggestions. Thanks also to Eduardo de Proença Barbosa for helping us with some molecular analysis. We thank Blanca Huertas for all the help she provided during the visitation of the Lepidoptera Collection. We also thank Daniel H. Janzen and Winifred Hallwachs for sharing the photos and information about the immature stages. JRAL thanks the Society of Systematic Biologists for the grant “Graduate Student Research Award (GSRA)” used to COI-sequencing; to the Museum of Comparative Zoology, Harvard University, for the “Ernst Mayr Travel Grants in Animal Systematics” used to visit the Lepidoptera Collection from NHMUK; the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES - 88882.382399/2019-01) and the Programa Institucional de Internacionalização (CAPES-PRINT 88887.474486/2020-00) for the research fellowships. RRS thanks the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP - 2020/09595-0). MMC and OHHM thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq - 302084/2017-7 and CNPq - 304849/2019-7) for the research fellowship. The authors also thank the Canadian Center for DNA Barcoding (CCDB) for molecular procedures.

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