Tanya Dapkey

Integration of DNA barcoding into an ongoing inventory of complex tropical biodiversity

Inventory of the caterpillars, their food plants and parasitoids began in 1978 for todays Area de Conservacion Guanacaste (ACG), in northwestern Costa Rica. This complex mosaic of 120 000 ha of conserved and regenerating dry, cloud and rain forest over 0–2000 m elevation contains at least 10 000 species of non‐leaf‐mining caterpillars used by more than 5000 species of parasitoids. Several hundred thousand specimens of ACG‐reared adult Lepidoptera and parasitoids have been intensively and extensively studied morphologically by many taxonomists, including most of the co‐authors. DNA barcoding — the use of a standardized short mitochondrial DNA sequence to identify specimens and flush out undisclosed species — was added to the taxonomic identification process in 2003. Barcoding has been found to be extremely accurate during the identification of about 100 000 specimens of about 3500 morphologically defined species of adult moths, butterflies, tachinid flies, and parasitoid wasps. Less than 1% of the species have such similar barcodes that a molecularly based taxonomic identification is impossible. No specimen with a full barcode was misidentified when its barcode was compared with the barcode library. Also as expected from early trials, barcoding a series from all morphologically defined species, and correlating the morphological, ecological and barcode traits, has revealed many hundreds of overlooked presumptive species. Many but not all of these cryptic species can now be distinguished by subtle morphological and/or ecological traits previously ascribed to ‘variation’ or thought to be insignificant for species‐level recognition. Adding DNA barcoding to the inventory has substantially improved the quality and depth of the inventory, and greatly multiplied the number of situations requiring further taxonomic work for resolution.

Nuclear genomes distinguish cryptic species suggested by their DNA barcodes and ecology

Significance Thirteen years of mitochondrial DNA barcoding of 15,000+ species of Lepidoptera and their parasitoids living in Area de Conservación Guanacaste, northwestern Costa Rica, indicate several thousand cases where barcodes combined with ecology suggest unrecognized cryptic species, substantially increasing species counts. Here, we show that the slightly different barcodes of three extremely similar parapatric–sympatric species of butterflies covary not only with ecology and subtle morphological traits but also with nuclear genomes—a finding that we predict will be commonplace and a method that we predict will be widely used. The barcodes of the century-old type specimens of Udranomia kikkawai from Venezuela reveal that this name applies to one of the three Costa Rican cryptic species; the others we describe as new. 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.

Lytopylus Förster (Hymenoptera, Braconidae, Agathidinae) species from Costa Rica, with an emphasis on specimens reared from caterpillars in Area de Conservación Guanacaste

Abstract Twelve species of Costa Rican Lytopylus are treated; these include all species reared from Lepidoptera caterpillars in Area de Conservación Guanacaste, Costa Rica, over 32 years of caterpillar inventory, as well as two species recorded in the literature as occurring in Costa Rica. Ten new species are described, i.e., Lytopylus bradzlotnicki, Lytopylus colleenhitchcockae, Lytopylus gregburtoni, Lytopylus jessicadimauroae, Lytopylus jessiehillae, Lytopylus mingfangi, Lytopylus rebeccashapleyae, Lytopylus robpringlei, Lytopylus sandraberriosae, Lytopylus vaughntani. The following species are transferred to Lytopylus: Metriosoma flavicalcar Enderlein 1920 to Lytopylus flavicalcar comb. n.; Bassus macadamiae Briceño and Sharkey 2000 to Lytopylus macadamiae comb. n.; Metriosoma bicarinatum Enderlein 1920 to Lytopylus bicarinatum comb. n.; Metriosoma brasiliense Enderlein 1920 to Lytopylus brasiliense comb. n.; Bassus tayrona Campos 2007 to Lytopylus tayrona comb. n.; Microdus femoratus Cameron 1887 to Lytopylus femoratus comb. n.; Microdus melanocephalus Cameron 1887 to Lytopylus melanocephalus comb. n.; Bassus pastranai Blanchard 1952 to Lytopylus pastranai comb. n.; Agathis nigrobalteata Cameron 1911 to Lytopylus nigrobalteatus comb. n. Two keys to species of Lytopylus are presented, one interactive and the other static.

Random Walk Modeling of Adult Leuctra ferruginea (Stonefly) Dispersal

Abstract Aquatic insects of the orders Ephemeroptera, Plecoptera and Trichoptera (EPT) are commonly used as indicators of freshwater quality. Recolonization of restored waterways by these pollution intolerant insects is frequently employed to measure the success of stream restoration efforts. Post restoration monitoring programs based on recolonization by EPT indicator species are typically generalized with little consideration for local conditions. Within Philadelphia urban development has resulted in a loss of unimpaired headwater streams that might serve as refugia of EPT colonists to replenish downstream populations through passive drift of larvae and ova. In this urban setting, recolonization may depend largely on overland flight by adults from other systems. In an attempt to predict the likelihood of overland dispersal by adult EPT within Philadelphia, PA, USA, a one year field study was conducted at the Schuylkill Center for Environmental Education (SCEE) in Northwest Philadelphia to obtain data on local EPT taxa and dispersal behavior. Adult EPT specimens belonging to 15 species were collected using sticky traps extending laterally from Meigs Run, a 1st order tributary to the Schuylkill River. Leuctra ferruginea (Plecoptera: Leuctridae) was selected as the study species. A random walk model was developed and calibrated to field observations to predict long distance dispersal probability. An individual-based Levy flight random walk model provided the best fit for field observations from Meigs Run, as well as supporting observations and theory proposed in existing literature.

Two new species of Erythromelana Townsend, 1919 (Diptera: Tachinidae) from Area de Conservación Guanacaste in northwestern Costa Rica

Abstract Background We describe two new species in the genus Erythromelana Townsend, 1919 from Area de Conservación Guanacaste (ACG) in northwestern Costa Rica. Both species were reared from wild-caughtcaterpillars of Eois spp. (Lepidoptera: Geometridae). We provide a concise description of each species using morphology, life history, molecular data, and photographic documentation. New information Erythromelana jimmychevezi Fleming & Wood sp. nov. Erythromelana glenriverai Fleming & Wood sp. nov.

Review of Lophomyra Schaus, 1911 (Lepidoptera, Noctuidae): a new combination and re-descriptions of species newly associated with ferns (Polypodiaceae)

Abstract Lophomyra Schaus, 1911 (Noctuidae) is the smaller of two noctuid genera originally described by Schaus that include species recently associated with ferns (Pteridophyta), in this case Polypodiaceae, as larval food plants. Following an examination of type material and reared specimens accompanied by DNA barcode data, Lophomyra is revised to include L.tacita Schaus, 1911, L.santista (Jones, 1914), and L.commixta (Schaus, 1914), comb. n., the last of which is transferred from Chytonidia Schaus, 1914 (= Leucosigma Druce, 1908). Lophomyra is characterized based on adult and larval morphology, especially that of the male genitalia. Structures associated with the valvae are discussed with reference to dissected and in situ images. Larvae of L.commixta and L.tacita are described from images, and the recorded food plants of both species are discussed in the context of known New World noctuid pteridivores.

A review of Leucosigma Druce, 1908: a newly discovered case of fern-feeding and descriptions of three new species (Lepidoptera, Noctuidae)

Abstract Chytonidia Schaus, 1914, is one of two noctuine genera originally described by Schaus that includes species recently found to feed on fern foliage (Pteridophyta) as larvae. By examining museum specimens, including type material and reared specimens accompanied by DNA barcode data, Chytonidia Schaus, 1914, syn. n. is synonymized with Leucosigma Druce, 1908, all currently recognized species are re-described, including males of three species described from female holotypes, and three new species are described: Leucosigmasolisae Goldstein, sp. n., Leucosigmapoolei Goldstein, sp. n., and L.schausi Goldstein, sp. n. Images of adults and, where available, larvae as well as dissected genitalia are presented, with a key to adults.

Nine new species of Uramya Robineau-Desvoidy (Diptera: Tachinidae) from Area de Conservación Guanacaste in northwestern Costa Rica, with a key to their identification

Abstract Background We describe nine new species in the genus Uramya Robineau-Desvoidy, 1830 from Area de Conservación Guanacaste (ACG) in northwestern Costa Rica. All species were reared from an ongoing inventory of wild-caught caterpillars spanning a variety of families (Lepidoptera: Erebidae; Limacodidae; Megalopygidae; Lasiocampidae and Dalceridae). Our study provides a concise description of each new species using morphology, life history, molecular data, and photographic documentation. In addition to the new species the authors provide a redescription the previously described Uramya sibinivora Guimarães, which was also collected within ACG during this study. We also provide a redescription of the genus, and a revised key to species of Uramya occurring in Central and South America. New information The following nine new species of Uramya, all authored by Fleming & Wood, are described: Uramya albosetulosa Fleming & Wood sp. nov., Uramya constricta Fleming & Wood sp. nov., Uramya contraria Fleming & Wood sp. nov., Uramya infracta Fleming & Wood sp. nov., Uramya lativittata Fleming & Wood sp. nov., Uramya lunula Fleming & Wood sp. nov., Uramya nitida Fleming & Wood sp. nov., Uramya pannosa Fleming & Wood sp. nov., and Uramya penicillata Fleming & Wood sp. nov. The following are proposed by Wood as new synonyms of Uramya: Olinda Townsend, syn. nov. and Procleonice Townsend, syn. nov. The following new combination is proposed as a result of the new synonymies: Uramya brasiliensis Macquart, comb. nov. Procleonice prolixa Townsend is synonymized under Uramya brevicauda Curran, syn. nov.

Five new species of Vibrissina Rondani (Diptera: Tachinidae) from Area de Conservación Guanacaste in Northwestern Costa Rica

Abstract Background We describe five new species in the genus Vibrissina Rondani from Area de Conservación Guanacaste (ACG). All species were reared from wild-caught sawfly larvae (Hymenoptera: Symphyta: Argidae and Tenthredinidae). We provide a morphological description of each species together with information on life history, molecular data, and photographic documentation. New information Five new species of Vibrissina Rondani: Vibrissina randycurtisi sp. n., V. randyjonesi sp. n., V. robertwellsi sp. n., V. danmartini sp. n., V. hallwachsorum sp. n.

Revision of the genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservación Guanacaste, Costa Rica, with a key to all species previously described from Mesoamerica

The genus Promicrogaster (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservación Guanacaste (ACG), Costa Rica, is revised. A key is provided to all new species as well as those previously described from Mesoamerica. A total of 21 species, all authored by Fernandez-Triana & Boudreault, are described as species nova: alexmartinezi, andreyvallejosi, brandondinartei, daniellopezi, daretrizoi, eddycastroi, eimyobandoae, fabiancastroi, fabriciocambroneroi, hillaryvillafuerteae, kevinmartinezi, kiralycastilloae, leilycastilloae, liagrantae, luismendezi, monteverdensis, naomiduarteae, pablouzagai, ronycastilloi, sebastiancambroneroi, tracyvindasae. A species previously described from India is considered as incertae sedis. Promicrogaster is considered to be restricted to the New World, with the vast majority of the species found in the Neotropics and a few extending north to the Nearctic. Almost 60% of the known species in ACG are found in cloud forests at over 1,000 m altitude. All of the verified and authenticated host records for Promicrogaster are from caterpillars living more deeply inside plant tissue than simply in rolled leaf structures – although no host data from ACG is available. JHR 50: 25–79 (2016) doi: 10.3897/JHR.50.8220 http://jhr.pensoft.net Copyright Jose Fernández-Triana et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. RESEARCH ARTICLE Jose Fernández-Triana et al. / Journal of Hymenoptera Research 50: 25–79 (2016) 26