Multi-surveyor detection-mark-redetection as a powerful tool for butterfly population monitoring in the pre-imaginal stage

1. For many elusive insect species, which are difficult to cover by standard monitoring schemes, innovative monitoring methods are needed to gain robust data on population trends. We suggest a monitoring of overwintering larvae for the endangered nymphalid butterfly Limenitis reducta. 2. We tested one removal and three detection-mark-redetection (DMR) approaches in a field study in the “Alb-Donau” region, Germany. We replaced movement of the study organisms by random movement of multiple different surveyors, and we examined the model assumption of equal detectability using simulations. 3. Our results indicate that multi-surveyor removal/DMR techniques are suitable for estimating abundance of overwintering L. reducta larvae. Detection probabilities varied with surveyor experience and the uncertainty of population estimates increased with a decrease in personnel expenditure. Estimated larval densities on a spruce clear-cut ranged between one and three individuals per 100 m². 4. We suggest a detection-mark-redetection (DMR) approach with three trained surveyors for the monitoring of L. reducta populations in the pre-imaginal stage. Besides L. reducta, the proposed method is likely to be suitable for other insect taxa with specific immobile life-stages and some sessile organisms, e.g. corals, elusive plants, or fungi.

Systematic Revision of a New Butterfly Genus, Cisandina Nakahara & Espeland, n. gen., with Descriptions of Three New Taxa (Lepidoptera: Nymphalidae: Satyrinae)

We here establish a new genus in the nymphalid butterfly subtribe Euptychiina, Cisandina Nakahara & Espeland, n. gen. to harbor five species hitherto placed within two polyphyletic genera, namely Magneuptychia Forster, 1964 and Euptychoides Forster, 1964. We compiled data from over 350 specimens in 17 public and private collections, as well as DNA sequence data for all relevant species, to revise the species-level classification of this new genus. According to our multi-locus molecular phylogeny estimated with the maximum likelihood approach, Cisandina lean. comb., Cisandina philippan. comb. & reinst. stat., Cisandina fidan. comb., Cisandina sanmarcosn. comb., and Cisandina trinitensisn. comb. are proposed as new taxonomic combinations, since these species are distantly related to the type species of Magneuptychia and Euptychoides and cannot reasonably be accommodated in any other genus. Lectotypes are designated for Papilio lea Cramer, 1777, Papilio junia Cramer, 1780, Euptychia philippa Butler, 1867, and Eupytchia fida Weymer, 1911. Two new species of Cisandinan. gen. are named and described herein, C. esmeralda Nakahara & Barbosa, n. sp. and C. castanya Lamas & Nakahara, n. sp., increasing the described species diversity of the genus to seven. The immature stages of C. castanyan. sp. and C. philippan. comb. & reinst. stat. are documented along with their natural hostplants, representing the first two species of the genus with known life history information. We describe a new subspecies, Cisandina fida directa Nakahara & Willmott, n. ssp., based on a limited number of specimens from southern Ecuador and central Peru. We were unable to obtain genetic data for the nominate race of C. fidan. comb., and thus, this taxonomic hypothesis is currently based solely on phenotypic characters.

An overview of “arginussa” species group of Memphis Hübner, [1819], with the description of a new species from Panama (Lepidoptera: Nymphalidae: Charaxinae)

Memphis Hübner, [1819] is a nymphalid butterfly genus exclusive to the Neotropics. It includes species with iridescent bluish or greenish coloration on the upper side of the wings while the underside is cryptic, resembling dead leaves. This paper aims to describe a remarkable new species, M. smalli Riley & Dias sp. nov., from the remote Atlantic slopes of Panama based on molecular and morphological analyses. We also review the taxonomy of species herein included in the “arginussa” species group based on distances analyses of DNA sequence data. The “arginussa” species group, as defined here, includes M. arginussa (Geyer, 1832), M. eubaena (Boisduval, 1870) stat. rest., M. onophis (Felder & Felder, 1861) stat. rest., M. lemons (Druce, 1877), M. neidhoeferi (Rotger, Escalante & Coronado, 1965), M. perenna (Godman & Salvin, [1884]), M. lankesteri (Hall, 1935) stat. rest., M. paulus Costa & Orellana, 2014, M. pithyusa (Felder, 1869), M. herbacea (Butler & Druce, 1872) and M. smalli sp. nov. Anaea pithyusa morena Hall, 1935 syn. nov. is recognized as a synonym of Nymphalis pithyusa Felder, 1869. The new species and its closest ally, M. herbacea, are illustrated, including characters of the head, labial palpus, wings, legs, male and female genitalia and their distribution map.

Conserved roles of Distal-less and spalt in regulating butterfly melanic patterns

Simple spot patterns are proposed to be ancestral to more complex eyespot patterns found on the wings of butterflies. Two genes, Distal-less (Dll) and spalt (sal), are known to be involved in two separate functions in establishing nymphalid butterfly eyespots: in the differentiation of their central signaling cells, or foci, necessary for eyespot ring formation, and in scale melanisation. However, little is known about the functions of these genes in the development of more primitive spot patterns. Here, we study the roles of Dll and sal in the development of spots and other melanic wing patterns of the Indian cabbage white, Pieris canidia, a pierid butterfly. We examined the expression pattern of both genes in developing larval and pupal wings and explored their functions using the CRISPR/Cas9 system. In P. canidia, both Dll and sal are expressed in the tips and along the margin of pupal wings, areas of future scale melanisation. Additionally, sal alone, is expressed in the future black spot patterns. CRISPR knockouts of Dll and sal showed that both genes are required for promoting melanic scales in the areas where they are expressed. Both Dll and sal also function as repressors of the pteridine pigment pathway. We conclude that both genes share a conserved role in promoting scale melanisation, across pierid and nymphalid butterflies, but are unlikely to be involved in differentiating spot centers. The genetic and developmental mechanisms that set up the location of spots and eyespots are likely distinct and independently evolved in each lineage.

First evidence for an Amazonian insect migration in the butterfly Panacea prola (Lepidoptera: Nymphalidae)

ABSTRACTInsect migrations rival those of vertebrates in terms of numbers of migrating individuals and even biomass, although instances of the former are comparatively poorly documented. This is especially true in the world’s tropics, which harbor the vast majority of Earth’s insect species. Understanding these mass movements is of critical and increasing importance as global climate and land use change accelerate and interact to alter the environmental cues that underlie migration, particularly in the tropics. Here, we provide the first evidence for an insect migration for the nymphalid butterfly Panacea prola in the Amazon, the world’s largest and most biodiverse rainforest that is experiencing a shifting climate and rapid forest loss.

Multiple roles for laccase2 in butterfly wing pigmentation, scale development, and cuticle tanning

ABSTRACTLepidopteran wing scales play important roles in a number of functions including color patterning and thermoregulation. Despite the importance of wing scales, however, we still have a limited understanding of the genetic mechanisms that underlie scale patterning, development, and coloration. Here we explore the function of the phenoloxidase-encoding gene laccase2 in wing and scale development in the nymphalid butterfly Vanessa cardui. Somatic deletion mosaics of laccase2 generated by CRISPR/Cas9 genome editing presented several distinct mutant phenotypes. Consistent with work in other non-lepidopteran insect groups, we observed reductions in melanin pigmentation and defects in cuticle formation. We were also surprised, however, to see distinct effects on scale development including complete loss of wing scales. This work highlights laccase2 as a gene that plays multiple roles in wing and scale development and provides new insight into the evolution of lepidopteran wing coloration.

Immature stages of Neptis reducta Fruhstorfer, 1908 (Lepidoptera, Nymphalidae, Neptini): morphology, ecology and food specialty

Neptis reducta Fruhsorfer, 1908 is currently recognized as a nymphalid butterfly found only in Taiwan and Thailand, with previously recorded larval hostplant and immature biology involving mis-identifications. The present study recognizes Aphananthe aspera (Cannabaceae) as the sole larval hostplant for N. reducta, and the nymphalid butterfly is the first known lepidopterous insect specialized on Aphananthe.

Molecular identification and phylogenetic relationships of seven Satyrinae butterflies in Bangladesh using Cytochrome c oxidase subunit I gene

The Satyrinae is a subfamily of Nymphalid butterfly, which is morphologically and ecologically the most diverse group, occurring in all habitats. In the present study, Cytochrome c oxidase subunit I (COI) gene of seven species of Satyrinae was sequenced, aligned, and used to construct phylogenetic trees. The molecular identification of these Satyrinae species was confirmed by comparing the related sequences in the National Center for Biotechnology Information (NCBI) GenBank. The base compositions of the COI sequences were 39.07% T, 16.44% C, 29.83% A, and 14.64% G, revealing a strong AT bias (68.9%). The sequence distance among Satyrinae species ranged from 0.09% to 0.18%. Phylogenetic trees were constructed by the neighbor-joining (NJ) and maximum likelihood (ML) methods, using Orthetrum sabina as an outgroup. Both trees had almost identical topologies. The sampled species in Satyrinae exhibited the following relationships: Melanitis leda + [(Mycalesis mineus+(Mycalesis gotama+Mycalesis anaxias)) + (Ypthima baldus + (Lethe chandica+Elymnias hypermnestra))], suggesting that M. leda might be distantly related with the rest of the Satyrinae species. This clustering result is almost identical to current traditional classification. This study confirms that the COI based DNA barcoding is an efficient method for the identification of butterflies including Satyrinae species and, as such, may further contribute effectively to biodiversity and evolutionary research. Jahangirnagar University J. Biol. Sci. 8(1): 67-74, 2019 (June)

Butterfly assemblages from Amazonian flooded forests are not more species-poor than from unflooded forests

The Amazonian flooded and upland forests harbour distinct assemblages of most taxonomic groups. These differences can be mainly attributed to flooding, which may affect directly or indirectly the persistence of species. Here, we compare the density, richness and composition of butterfly assemblages in vaórzea and terra firme forests, and evaluate whether terrain elevation and flooding can be used to predict the assemblage structure. We found that the total abundance and number of species per plot is higher in vaórzea than in terra firme forests. Vaórzea assemblages showed a higher dominance of abundant species than terra firme assemblages, in which low-flying Haeterini butterflies had higher abundance. After standardizing species richness by sample size and/or coverage, species richness estimates for vaórzea and terra firme forests were similar. There was strong turnover in species composition across vaórzea and terra firme forests associated with terrain elevation, most likely due to differences in the duration of flooding. Despite a smaller total area, less defined vegetation strata, more frequent disturbances and the younger geological age of floodplain forests, Nymphalid butterfly assemblages are not more species poor there than in unflooded forests.