RESUMO

Genomic sequencing and analysis of holotypes from the MIZA collection (Maracay, Venezuela) and their comparison with other species and their type specimens advances our understanding of their taxonomy. Jemadia demarmelsi Orellana, [2010] is confirmed as a species-level taxon and its female is genetically verified. The following are species-level taxa, not subspecies: Amenis pedro O. Mielke & Casagrande, 2022, stat. nov. (not Amenis pionia (Hewitson, 1857)) and Jemasonia sosia (Mabille, 1878), stat. rest. (not Jemasonia hewitsonii (Mabille, 1878)). Amenis ponina rogeri Orellana, [2010], stat. nov. and Jemasonia pater ortizi (Orellana, [2010]), stat. nov. are subspecies, not species. Jemadia pseudognetus imitator (Mabille, 1891), comb. nov. (not Jemadia hospita (Butler, 1877)) and Damas cervelina Orellana & Costa, 2019, comb. nov. (not Megaleas Godman, 1901) are new combinations.

RESUMO

Jonaspyge elizabethae n. sp. is described from southwestern Honduras. It is similar to the other two Jonaspyge O. Mielke, 2002 species in having metallic dark-blue wings with purple sheen, crenulate hindwing outer margin, and black body with orange palpi and an orange abdomen tip. It is diagnosed by bright-orange (instead of white) fringes and dark (instead of orange) cheeks. Genomic sequence analysis of Jonaspyge reveals that it is a close relative of Jonaspyge jonas (C. Felder & R. Felder, 1859) and Jonaspyge tzotzili (H. Freeman, 1969), differing from them by 5.3% in the COI DNA barcode. This new, third species of Jonaspyge is the most divergent member of the genus.

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Emesis eleanorae Gallardo & Grishin n. sp. is described from western Honduras. It differs from other species of Emesis Fabricius, 1807 in having a row of prominent iron-gray crescent-shaped postdiscal spots on both wings above, outlined by paler areas basad and mirrored as merlot-colored spots below, with the largest by the forewing costa, and in its females being bright golden-orange in color. Genomic sequence analysis of Emesis reveals that the new species belongs to the subgenus Aphacitis Hübner, [1819] and is sister to the clade containing Emesis diogenia Prittwitz, 1865 and Emesis heteroclita Stichel, 1929, and the clade of these three species is sister to Emesis vulpina Godman & Salvin, 1886.

Emesis eleanorae Gallardo & Grishin n. sp. se describe desde el Occidente de Honduras. Se diferencia de otras especies de Emesis Fabricius, 1807 por tener una fila de prominentes puntos postdiscales en forma de media luna, color gris-hierro en la parte de arriba de ambas alas, delineadas por áreas más pálidas y reflejadas debajo como manchas de color merlot, las cuales son más grandes en la costa delantera, y en las hembras son de color dorado-anaranjado brillante. El análisis de la secuencia genómica de Emesis revela que la nueva especie pertenece al subgénero Aphacitis Hübner, [1819] y es pariente del clado de Emesis diogenia Prittwitz, 1865 y Emesis heteroclita Stichel, 1929, y el clado de estas tres especies son parientes de Emesis vulpina Godman y Salvin, 1886.

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Recent taxonomic studies on the genus Wahydra Steinhauser, 1991, have described five new species in the past two years, from high elevations in the Andes Mountains. A markedly distinct species of Wahydra is herein described and illustrated based on a single male specimen from Ecuadorian Andes, Wahydra graslieae A. Warren, Carneiro Dolibaina, sp. nov. The new species is compared with other species of Wahydra, as well as with the somewhat similar species Lerema viridis (Bell, 1942) and Tigasis viridenex (Weeks, 1901).

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Oxynetra aureopectasp. n. is described from the Sierra Madre Oriental of east-central Mexico. Visually similar to Mesoamerican O. hopfferi Staudinger, 1888 in having five orange bands on the abdomen above, it is diagnosed by orange forecoxae and palpi beneath, narrower forewing hyaline bands and a prominent 6% difference in the COI DNA barcode sequence. It is the northernmost representative of the hopfferi species group that also includes O. stangelandi Grishin & Burns, 2013, characterized by a single-banded abdomen and currently known only from the Area de Conservación Guanacaste in northwestern Costa Rica. Both O. hopfferi and O. stangelandi possess white forecoxae and ventral palpi. This new discovery brings the total number of Oxynetra C. & R. Felder, 1862 species to five.

Resumen Oxynetra aureopectasp. n. se describe de la Sierra Madre Oriental, en el centro-este de México. Visualmente similar a la especie mesoamericana O. hopfferi Staudinger, 1888 en tener cinco bandas de color naranja en el abdomen anterior, se diagnostica por forecoxae y palpos naranja debajo, bandas hialinas más estrechas en la ala anterior, y una diferencia destacada de 6% en la secuencia de código de barras de ADN COI. Es el representante más septentrional del grupo de especies hopfferi que también incluye O. stangelandi Grishin & Burns, 2013, que se caracteriza por una banda en el abdomen y actualmente conocida solamente desde el Área de Conservación Guanacaste en el noroeste de Costa Rica. Tanto O. hopfferi y O. stangelandi poseen forecoxae y cara ventral de palpos blancos. Este nuevo descubrimiento eleva el número total de especies de Oxynetra C. & R. Felder a cinco.

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DNA sequencing brings another dimension to exploration of biodiversity, and large-scale mitochondrial DNA cytochrome oxidase I barcoding has exposed many potential new cryptic species. Here, we add complete nuclear genome sequencing to DNA barcoding, ecological distribution, natural history, and subtleties of adult color pattern and size to show that a widespread neotropical skipper butterfly known as Udranomia kikkawai (Weeks) comprises three different species in Costa Rica. Full-length barcodes obtained from all three century-old Venezuelan syntypes of U. kikkawai show that it is a rainforest species occurring from Costa Rica to Brazil. The two new species are Udranomia sallydaleyae Burns, a dry forest denizen occurring from Costa Rica to Mexico, and Udranomia tomdaleyi Burns, which occupies the junction between the rainforest and dry forest and currently is known only from Costa Rica. Whereas the three species are cryptic, differing but slightly in appearance, their complete nuclear genomes totaling 15 million aligned positions reveal significant differences consistent with their 0.00065-Mbp (million base pair) mitochondrial barcodes and their ecological diversification. DNA barcoding of tropical insects reared by a massive inventory suggests that the presence of cryptic species is a widespread phenomenon and that further studies will substantially increase current estimates of insect species richness.

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Two species of hairstreak butterflies from the genus Calycopis are known in the United States: C. cecrops and C. isobeon Analysis of mitochondrial COI barcodes of Calycopis revealed cecrops-like specimens from the eastern US with atypical barcodes that were 2.6% different from either USA species, but similar to Central American Calycopis species. To address the possibility that the specimens with atypical barcodes represent an undescribed cryptic species, we sequenced complete genomes of 27 Calycopis specimens of four species: C. cecrops, C. isobeon, C. quintana and C. bactra Some of these specimens were collected up to 60 years ago and preserved dry in museum collections, but nonetheless produced genomes as complete as fresh samples. Phylogenetic trees reconstructed using the whole mitochondrial and nuclear genomes were incongruent. While USA Calycopis with atypical barcodes grouped with Central American species C. quintana by mitochondria, nuclear genome trees placed them within typical USA C. cecrops in agreement with morphology, suggesting mitochondrial introgression. Nuclear genomes also show introgression, especially between C. cecrops and C. isobeon About 2.3% of each C. cecrops genome has probably (p-value < 0.01, FDR < 0.1) introgressed from C. isobeon and about 3.4% of each C. isobeon genome may have come from C. cecrops. The introgressed regions are enriched in genes encoding transmembrane proteins, mitochondria-targeting proteins and components of the larval cuticle. This study provides the first example of mitochondrial introgression in Lepidoptera supported by complete genome sequencing. Our results caution about relying solely on COI barcodes and mitochondrial DNA for species identification or discovery.

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Two new species of Clito from the C. littera group are described: C. palotschka, sp. nov. from the Chocó region in Ecuador is the most distinctive in the group, and C. litteroides, sp. nov. from Venezuela is superficially more similar to C. littera differing mostly in the shape of male genitalic valvae. Comparative analyses of male genitalia and wing patterns suggest that C. anda Evans, 1953, new status; and C. nebulosa (Draudt, 1924), reinstated status are species-level taxa. A lectotype for Telemiades littera Mabille, 1877 is designated to ensure nomenclatural stability. Primary type specimens of all seven species in the C. littera group are illustrated with photographs, and an identification key to males is given. 

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Hermeuptychia intricata Grishin, sp. n. is described from the Brazos Bend State Park in Texas, United States, where it flies synchronously with Hermeuptychia sosybius (Fabricius, 1793). The two species differ strongly in both male and female genitalia and exhibit 3.5% difference in the COI barcode sequence of mitochondrial DNA. Setting such significant genitalic and genotypic differences aside, we were not able to find reliable wing pattern characters to tell a difference between the two species. This superficial similarity may explain why H. intricata, only distantly related to H. sosybius, has remained unnoticed until now, despite being widely distributed in the coastal plains from South Carolina to Texas, USA (and possibly to Costa Rica). Obscuring the presence of a cryptic species even further, wing patterns are variable in both butterflies and ventral eyespots vary from large to almost absent. To avoid confusion with the new species, neotype for Papilio sosybius Fabricius, 1793, a common butterfly that occurs across northeast US, is designated from Savannah, Georgia, USA. It secures the universally accepted traditional usage of this name. Furthermore, we find that DNA barcodes of Hermeuptychia specimens from the US, even those from extreme south Texas, are at least 4% different from those of H. hermes (Fabricius, 1775)-type locality Brazil: Rio de Janeiro-and suggest that the name H. hermes should not be used for USA populations, but rather reserved for the South American species. This conclusion is further supported by comparison of male genitalia. However, facies, genitalia and 2.1% different DNA barcodes set Hermeuptychia populations in the lower Rio Grande Valley of Texas apart from H. sosybius. These southern populations, also found in northeastern Mexico, are described here as Hermeuptychia hermybius Grishin, sp. n. (type locality Texas: Cameron County). While being phylogenetically closer to H. sosybius than to any other Hermeuptychia species, H. hermybius can usually be recognized by wing patterns, such as the size of eyespots and the shape of brown lines on hindwing. "Intricate Satyr" and "South Texas Satyr" are proposed as the English names for H. intricata and H. hermybius, respectively.