Consequences of the arms race between <Emphasis Type="Italic">Maculinea teleius</Emphasis> social parasite and <Emphasis Type="Italic">Myrmica</Emphasis> host ants for myrmecophilous butterfly conservation

The arms race between Maculinea butterflies and Myrmica host ants leads to local host-parasite adaptations. In our study, we assessed whether sympatric and allopatric Myrmica scabrinodis populations exhibit behavioural differences towards Maculinea teleius larvae during the adoption-period when butterfly larvae need to be taken inside the Myrmica nest. The second aim was to assess the butterfly survival rate inside ant colonies from different populations. We used one sympatric host population and three allopatric populations: one infested by M. teleius and two uninfested populations. We found that ants from the sympatric population showed a higher number of positive behaviours toward M. teleius larvae during adoption than ants from the allopatric populations. There were no differences in the number of inspection or negative behaviour events. The survival of butterfly larvae was highest inside sympatric host colonies and differed from the survival of M. teleius reared by ants from the allopatric, uninfested populations. No difference was found for the survival rate of M. teleius raised by infested, allopatric host colonies compared to sympatric host populations. Our results suggest the lack of behavioural counter-adaptations of local hosts of M. teleius that more easily adopt and rear butterfly caterpillars compared to naive M. scabrinodis colonies. Our results may also have implications for Maculinea butterfly conservation, especially for reintroduction programmes. We suggest that the existence of behavioural host defences should be checked for the source host population, as well as for the Myrmica population from the reintroduction site. It may also be reasonable to introduce several Myrmica host colonies from the source butterfly host population.     

Patch occupancy and habitat of the hops azure (<Emphasis Type="Italic">Celastrina</Emphasis><Emphasis Type="Italic">humulus</Emphasis>), a rare North American endemic butterfly: insights for monitoring and conservation

The hops azure (Celastrina humulus Scott & D. Wright 1998) is a rare butterfly found along the Front Range of Colorado. Data on the prevalence of the butterfly and its preferred habitats are lacking. To describe the habitat of C. humulus at the southern part of its range, explore what factors impact C. humulus detectability, and estimate C. humulus habitat use along a riparian area known to support it, we conducted an occupancy analysis along the largest riparian system at the U.S. Air Force Academy (USAFA) in Colorado, USA. We used environmental and site-specific covariates to model the probability of detection and the probability of occupancy. Probability of detecting C. humulus was influenced by the amount of cloud cover during sampling, while the probability of occupancy was influenced by the total area of the host plant (wild hops) at the site. Probability of detection was higher during the first visit (69 %) than the second visit (64 %), and the probability of occupancy was higher (77 %) than assumed (30 %). Despite the host plant being patchily distributed throughout the butterfly’s range, the riparian areas at USAFA had a high prevalence of both wild hops and C. humulus. We use the project findings to develop future sampling efforts for the butterfly along tributaries at other locales within the butterfly’s range.     

Garden varieties: How attractive are recommended garden plants to butterflies?

One way the public can engage in insect conservation is through wildlife gardening, including the growing of insect-friendly flowers as sources of nectar. However, plant varieties differ in the types of insects they attract. To determine which garden plants attracted which butterflies, we counted butterflies nectaring on 11 varieties of summer-flowering garden plants in a rural garden in East Sussex, UK. These plants were all from a list of 100 varieties considered attractive to British butterflies, and included the five varieties specifically listed by the UK charity Butterfly Conservation as best for summer nectar. A total of 2659 flower visits from 14 butterfly and one moth species were observed. We performed a principal components analysis which showed contrasting patterns between the species attracted to Origanum vulgare and Buddleia davidii. The “butterfly bush” Buddleia attracted many nymphalines, such as the peacock, Inachis io, but very few satyrines such as the gatekeeper, Pyronia tithonus, which mostly visited Origanum. Eupatorium cannibinum had the highest Simpson’s Diversity score of 0.75, while Buddleia and Origanum were lower, scoring 0.66 and 0.50 respectively. No one plant was good at attracting all observed butterfly species, as each attracted only a subset of the butterfly community. We conclude that to create a butterfly-friendly garden, a variety of plant species are required as nectar sources for butterflies. Furthermore, garden plant recommendations can probably benefit from being more precise as to the species of butterfly they attract.     

Occupancy modeling of <Emphasis Type="Italic">Parnassius clodius</Emphasis> butterfly populations in Grand Teton National Park,Wyoming

Estimating occupancy patterns and identifying vegetation characteristics that influence the presence of butterfly species are essential approaches needed for determining how habitat changes may affect butterfly populations in the future. The montane butterfly species, Parnassius clodius, was investigated to identify patterns of occupancy relating to habitat variables in Grand Teton National Park and Bridger-Teton National Forest, Wyoming, United States. A series of presence–absence surveys were conducted in 2013 in 41 mesic to xeric montane meadows that were considered suitable habitat for P. clodius during their flight season (June–July) to estimate occupancy (ψ) and detection probability (p). According to the null constant parameter model, P. clodius had high occupancy of ψ?=?0.78?±?0.07 SE and detection probability of p?=?0.75?±?0.04 SE. In models testing covariates, the most important habitat indicator for the occupancy of P. clodius was a strong negative association with big sagebrush (Artemisia tridentata; β = ??21.39?±?21.10 SE) and lupine (Lupinus spp.; β?=???20.03?±?21.24 SE). While P. clodius was found at a high proportion of meadows surveyed, the presence of A. tridentata may limit their distribution within montane meadows at a landscape scale because A. tridentata dominates a large percentage of the montane meadows in our study area. Future climate scenarios predicted for high elevations globally could cause habitat shifts and put populations of P. clodius and similar non-migratory butterfly populations at risk.     

Observations on the ecology of the endangered butterfly <Emphasis Type="Italic">Zerynthia cassandra</Emphasis> in a protected area of Northern Italy

Zerynthia cassandra is a butterfly endemic to the Italian peninsula. It was recently recognized as a sister species of Zerynthia polyxena. Z. cassandra is oligophagous as it feeds on a very restricted number of toxic plants belonging to the genus Aristolochia: Aristolochia pallida and A. rotunda. The paper reports the results of a research on the ecology of this species which was studied in the natural reserve "Stagni di Lungavilla" (Northern Italy) where the butterfly was living in a small patch of grassy vegetation bordered by a ditch at the margins of a pond. Z. cassandra develops one generation a year. Oviposition occurred in April. Eggs were mainly laid on the lower side of A. rotunda blades. The height of eggs from the ground depended on climatic trends, which affected the host plant phenology. Eggs mortality was mainly due to consumption of leaves by snails (Cepaea nemoralis) and slugs (Arion sp.). Larvae completed their development from April to May, while chrysalises were the aestivating and overwintering stage. Females did not lay eggs on host plants when they were part of the undergrowth. A decline of the butterfly due to habitat changes was recorded during the research (2001–2005). According to information coming from local butterfly watchers the butterfly used to be quite common in the natural reserve until 70’s. Its decline was caused by agricultural intensification and the natural evolution of vegetation towards woodlands, which gradually reduced the host plant density, so making the habitat unsuitable and causing the local extinction of the butterfly.     

Effects of grass-specific herbicides on butterflies: an experimental investigation to advance conservation efforts

Encroachment by invasive plants is a leading threat to rare butterflies. Restoration plans increasingly recommend herbicides to control invasive plants within butterfly habitats. Few studies address the effects of these herbicides on at-risk butterflies. The effects of two graminicides (fluazifop-p-butyl and sethoxydim) and a surfactant (Preference^®) were evaluated on Icaricia icarioides blackmorei and Pieris rapae. The effects on butterfly larvae were assessed by mimicking recommended timing and mixture rates of field applications. Differences in survival to adult eclosure, development time, biomass, sex ratio and adult morphology were assessed. Survival of P. rapae was reduced by 32% with sethoxydim and 21% with fluazifop-p-butyl. Wing size and pupal weights of P. rapae were reduced by herbicide treatments. Icaricia icarioides blackmorei experienced a 21% reduction in development time from the date of treatment to eclosure. These results highlight the importance of careful consideration in the use of herbicides in habitats harboring at-risk butterfly populations.     

The effects of introduced vespid wasps (<Emphasis Type="Italic">Vespula germanica</Emphasis> and <Emphasis Type="Italic">V. vulgaris</Emphasis>) on threatened native butterfly (<Emphasis Type="Italic">Oreixenica ptunarra</Emphasis>) populations in Tasmania

Introduced vespid wasps (Vespula germanica and V. vulgaris) are highly efficient predators of native invertebrates. They have the potential to reduce populations of threatened species and change ecosystem dynamics, yet their impact is largely unknown in Australia. The introduction of vespid wasps has coincided with a decline in numbers of threatened Ptunarra brown butterflies (Oreixenica ptunarra) in Tasmania, Australia. The Ptunarra brown butterfly is endemic to Tasmania, where its habitat has been fragmented by clearance for agriculture and forestry. Local extinctions of the species were previously thought to be principally due to its inability to fly the long distances between habitat patches in this disjointed landscape. We investigate the importance of the new threat of vespid wasp predation in the decline of O. ptunarra in the highland grasslands of northwest Tasmania. Numbers of O. ptunarra analysed over a period of 15 years dramatically declined after the arrival of vespid wasps. Wasp control was trialled to determine whether it affected butterfly numbers. Current control methods decreased wasp numbers considerably, resulting in a small increase in butterfly numbers, indicating that wasp predation is keeping O. ptunarra at low densities. Without ongoing conservation measures, it is likely that butterfly numbers will stay low, potentially leading to genetic bottlenecks and more local extinctions. An increase in the intensity of wasp control, in combination with other conservation management methods, is required for the protection and recovery of O. ptunarra.     

Fen meadows on the move for the conservation of <Emphasis Type="Italic">Maculinea (Phengaris) teleius</Emphasis> butterflies

In the Netherlands, a single population of the obligate myrmecophilic butterfly Maculinea (Phengaris) teleius has survived on only 3 ha of habitat for more than 25 years, whereas at least 40 ha of habitat are thought to be required for a sustainable metapopulation. Therefore, 170 ha of farmland is being restored to wet meadows within a LIFE?+?project by large-scale soil excavation and hay inoculation. For successful restoration, the habitat requirements of the butterfly, with Sanguisorba officinalis as host plant and its particular life cycle as parasite of the ant species Myrmica scabrinodis, have to be taken into account. We tested whether colonization of nests of this ant species in the restoration areas is facilitated by translocation of sods collected from fen meadows. We divided 54 sods, each sized 1 m², randomly over six patches and measured vegetation development and ant presence in the sods and surrounding control plots for 2 years. In the first summer, significantly more Myrmica ants were found in the transplanted sods in comparison to the surrounding area. Herb cover had a significant positive effect on Myrmica ant presence while it did not affect the presence of the pioneer ant species Lasius niger. In the second year, Myrmica ants were found in the surrounding control plots as well. This study contributes to the knowledge-base required for the design of restoration projects aimed at expanding the habitat of the critically endangered butterfly Maculinea (Phengaris) teleius.     

Prenylated acetophenones from Melicope obscura and Melicope obtusifolia ssp. obtusifolia var. arborea and their distribution in Rutaceae

Four prenylated acetophenones 2,6-dihydroxy-4-geranyloxyacetophenone (1), 4-geranyloxy-2,6,β-trihydroxyacetophenone (2), 2,6-dihydroxy-4-geranyloxy-3-prenylacetophenone (3), and 4-geranyloxy-3-prenyl-2,6,β-trihydroxyacetophenone (4) have for the first time been isolated from Melicope obscura (1 and 2) and Melicope obtusifolia ssp. obtusifolia var. arborea (3 and 4). The distribution of prenylated acetophenones in Rutaceae is reviewed and the results, including the new records, indicate that prenylated acetophenones are valuable as chemotaxonomic markers for the subfamily Rutoideae, tribe Xanthoxyleae sensu Engler.     

Species distribution model for the ‘Northern’ Oak hairstreak (<Emphasis Type="Italic">Satyrium favonius ontario</Emphasis>) with comments on its conservation status in the northeastern United States

Satyrium favonius ontario: (W. H. Edwards) (Lepidoptera: Lycaenidae) is considered to be a rare butterfly in the northeastern United States. It receives legal protection in the state of Massachusetts as a Species of Special Concern. We studied the ecology and natural history of a colony of S. f. ontario at Great Blue Hills Reservation in Canton, Massachusetts. In addition, we assembled a database of confirmed S. f. ontario occurrences (n?=?362) and used this along with climate and oak abundance data to build a species distribution model for the northeastern portion of the butterfly’s range in the United States. The model predicts that essentially the entirety of southern New England is suitable for the species, and thus its modeled distribution extends well north of all documented colonies/localities. Just two climate variables, precipitation seasonality and minimum temperature of the coldest month, explained 95% of the model and largely determined relative suitability predictions. We make the case that the hairstreak is a canopy-dwelling insect that sporadically makes ground-level visits, and that its assumed regional rarity is due to detection difficulties rather than demographic rarity. While the butterfly may be imperiled and worthy of legal protection in portions of its range, we question the validity of population estimates and necessity of conservation efforts based on ground-level adult sightings, and recommend larval sampling using burlap bands as a more reliable method to census this butterfly. We also discuss the possibility that other Satyrium and more distantly related hairstreaks (e.g., Callophrys hesseli and Parrhasius m-album) may be additional examples of temperate, canopy-based butterflies.     

<Emphasis Type="Italic">Empruthotrema longipenis</Emphasis> n. sp. (Monogenoidea: Monocotylidae: Merizocotylinae) from the olfactory sacs of the smooth butterfly ray <Emphasis Type="Italic">Gymnura micrura</Emphasis> (Bloch &amp; Schneider) (Myliobatiformes: Gymnuridae) in the Gulf of Mexico

A new species of Empruthotrema Johnston & Tiegs, 1922 is described based on specimens collected from the olfactory sacs of smooth butterfly rays Gymnura micrura (Bloch & Schneider) captured in Mobile Bay (northcentral Gulf of Mexico), Alabama, USA. Empruthotrema longipenis n. sp. is most similar to the type-species Empruthotrema raiae (MacCallum, 1916) Johnston & Tiegs, 1922 by having 12 marginal and two interhamular loculi with members of haptoral hook pair 1 located midway along the periphery of each interhamular loculus and those of hook pair 2 located at the marginal termini of the bilateral septa flanking the interhamular loculi. Empruthotrema longipenis n. sp. differs from E. raiae by having a much longer male copulatory organ and from its remaining congeners by the sinistral and extracecal ejaculatory bulb flanking the pharynx, the number of interhamular and marginal septa, and the distribution of hook pairs 1 and 2 along the haptoral margin. This is the first report of a monocotylid from the smooth butterfly ray and from Mobile Bay. The diversity of haptoral morphotypes among the currently accepted species of Empruthotrema is detailed and discussed in the context of monophyly of the genus.     

Absolute configuration of tropane alkaloids from Schizanthus species by vibrational circular dichroism

The absolute configuration (AC) of 6β-hydroxy-3α-senecioyloxytropane (1), 3α-hydroxy-6β-tigloyloxytropane (2), 3α-hydroxy-6β-senecioyloxytropane (3), and 3α-hydroxy-6β-angeloyloxytropane (4) was assigned as (1R,3R,5S,6R) using density functional theory (DFT) calculations at the B3LYP/DGDZVP level of theory in combination with experimental vibrational circular dichroism (VCD) measurements and comparison with the spectra of similar tropanes. The AC of 1 followed from a sample isolated from Schizanthus grahamii, while those of the mixture of 2 and 3, isolated from the same source, were determined by comparing the VCD measurement to a weighted calculation of the individual VCD spectra according to a 69:31 ratio of 2:3 determined by ¹H NMR signal integration. In turn, Schizanthus pinnatus provided a 7:3 mixture of 1:4 whose AC was determined using the experimental VCD absorptions in the 1150-950 cm⁻¹ spectral region which were compared with those observed for 1-3 and with those described for other 3α,6β-tropanediol derivatives.     

Sesquiterpene constituents from the essential oils of the liverworts Mylia taylorii and Mylia nuda

The essential oils and extracts of Mylia taylorii and M. nuda were investigated by gas chromatography, mass spectrometry, NMR spectroscopy and chemical correlations. Beside several known compounds 13 new constituents including three new carbon skeletons could be identified. Four hydrocarbons with a molecular formula of C₁₅H₂₂ (m/z 202) were identified as myli-4(15)-ene (1), aromadendra-1(10),4(15)-diene (19), aromadendra-4,10(14)-diene (20) and aromadendra-4,9-diene (21). Three oxaspiro-compounds were identified as 7-epi-bourbon-3-en-5,11-oxide (22), guai-3,10(14)-dien-5,11-oxide (23) and guai-3,9-dien-5,11-oxide (24). The absolute configuration of myli-4(15)-en-3-one (5) could be established by chemical correlation. Together with α-taylorione (7) the corresponding 6,11-seco-compound taylopyran (25) with a new carbon skeleton was identified which serves as a precursor to taylocyclane (26) and taylofuran (27). Taynudol (28) contains a new carbon skeleton with a cyclobutenyl structure.     

Phomoeuphorbins A-D, azaphilones from the fungus Phomopsis euphorbiae

Four azaphilones, named phomoeuphorbins A-D (1-4) were isolated from cultures of Phomopsis euphorbiae, an endophytic fungus isolated from Trewia nudiflora. Structures of 1-4 were established on the basis of spectroscopic analyses, including application of 2D NMR spectroscopic techniques. Phomoeuphorbins A and C exhibited very weak inhibitory activities against HIV replication in C8166 cells in vitro.     

Unique wing scale photonics of male Rajah Brooke’s birdwing butterflies

Background

Ultrastructures in butterfly wing scales can take many shapes, resulting in the often striking coloration of many butterflies due to interference of light. The plethora of coloration mechanisms is dazzling, but often only single mechanisms are described for specific animals.

Results

We have here investigated the male Rajah Brooke’s birdwing, Trogonoptera brookiana, a large butterfly from Malaysia, which is marked by striking, colorful wing patterns. The dorsal side is decorated with large, iridescent green patterning, while the ventral side of the wings is primarily brown-black with small white, blue and green patches on the hindwings. Dense arrays of red hairs, creating a distinct collar as well as contrasting areas ventrally around the thorax, enhance the butterfly’s beauty. The remarkable coloration is realized by a diverse number of intricate and complicated nanostructures in the hairs as well as the wing scales. The red collar hairs contain a broad-band absorbing pigment as well as UV-reflecting multilayers resembling the photonic structures of Morpho butterflies; the white wing patches consist of scales with prominent thin film reflectors; the blue patches have scales with ridge multilayers and these scales also have centrally concentrated melanin. The green wing areas consist of strongly curved scales, which possess a uniquely arranged photonic structure consisting of multilayers and melanin baffles that produces highly directional reflections.

Conclusion

Rajah Brooke’s birdwing employs a variety of structural and pigmentary coloration mechanisms to achieve its stunning optical appearance. The intriguing usage of order and disorder in related photonic structures in the butterfly wing scales may inspire novel optical materials as well as investigations into the development of these nanostructures in vivo.

     

Synthesis, structures and ligating properties of 2,6-bis(phosphino)thiophenol derivatives

New t-butyl-aryl thioethers where the aryl group is 2,6-bis(phosphino)phenyl have been synthesized. The syntheses were completed via sequential ortho-lithiations of t-butylphenylsulfide, followed by chlorophosphine (ClPR₂) quenches; symmetric (2,6-bis(diphenylphosphino)phenyl, (4a)) and unsymmetric (2-diisopropylphosphino-6-diphenylphosphino)phenyl, (4b) aryl groups were obtained. Treatment of 4a with Li or Na naphthalenide yielded 2,6-bis(diphenylphosphino)thiophenol 5. Reactions of 4a or 5 with NiCl₂ · 6H₂O yielded nickel bis(phosphinothiophenolate) 6. Compounds 4a,b, 5 and 6 were characterized by ¹H and ³¹P NMR, and by mass-spectrometry. In addition, 4a, 5 and 6 were characterized by single crystal X-ray diffraction methods.     

Determination of the hydroxycinnamate profile of 12 members of the Asteraceae family

The hydroxycinnamates of the leaves of 12 plants of the Astreraceae family, Achillea millefolium, Arnica montana, Artemesia dracunculus, Cichorium intybus, Cnicus benedictus, Cynara scolymus, Echinops humilis, Inula helenium, Lactuca sativa, Petasites hybridus, Solidago virgaurea, and Tanacetum parthenium were investigated qualitatively by LC-MSⁿ. Thirty-nine chlorogenic acids were detected and all characterized to regioisomeric level on the basis of their fragmentation pattern in the tandem MS spectra, most of them for the first time from these sources with two of them previously not reported in nature. Both chlorogenic acids based on trans and cis-cinnamic acid substituents were identified. Assignment to the level of individual regioisomers was possible for seven caffeoylquinic acids (1-7), 11 dicaffeoylquinic acids (17-27), six feruloylquinic acids (9-14), two p-coumaroylquinic acids (15-16), two caffeoyl-feruloylquinic acids (28 and 29), four caffeoyl-p-coumaroylquinic acids (30-33), three dicaffeoyl-succinoylquinic acids (34-36), two dicaffeoyl-methoxyoxaloylquinic acids (37 and 38), and one tricaffeoylquinic acid (39). Furthermore, one caffeoylshikimic acid (40), one caffeoyltartaric acid (41), three dicaffeoyltartaric acids (42-44), and three caffeoyl-feruloyltartaric acids (45-47) were detected and shown to possess characteristic tandem MS spectra and were tentatively assigned on the basis of their retention time and previously developed hierarchical keys.     

2-(1-(Dimethylamino)ethyl)phenylpalladium(II) complexes 5-functionalized with fluorous silyl tails

New fluorous-organometallics based on the chiral ligand α-methyl-N,N-dimethylbenzylamine (TMBA) were prepared by treatment of fluorous silyl bromide reagents with in situ 4-lithiated TMBA to give fluorous N,N-dimethyl(α-methyl-4-trialkylsilylbenzyl)amine ligands 1a-1c that vary in the number of fluorous tails attached to the Si atom. Ligands 1a-1c were successfully cyclo-palladated by treatment with Pd(OAc)₂/LiCl in methanol to furnish the corresponding chloride-bridged dimeric arylpalladium(II) complexes 2a-2c in good yields. The latter derivatives could be converted into monomeric Lewis-base adducts by complexation with pyridine (3a-3c), or triphenylphosphine (4a-4c). The crystal structure of triphenylphosphine complex 4a has been elucidated. To probe their fluorophilicity, the partition coefficient of each of the derivatives in the fluorous biphasic solvent (FBS) system perfluoromethylcyclohexane/n-octane has been determined.     

Oxygenated verticillene derivatives from Bursera suntui

Medium polarity fractions of the hexane extracts of the stems of Bursera suntui afforded six previously known (1-6) and four hitherto unknown verticillane derivatives: (1S,3Z,7S,8S,11S,12S)-(+)-7,8-epoxyverticill-3-en-12,20-diol (7), (1S,3Z,7S,8S,11S,12S)-(+)-7,8-epoxyverticill-3-en-12,20-diol 20-acetate (8), (1S,3Z,7S,11S,12S)-(+)-verticilla-3,8(19)-dien-7,12,20-triol (9), and (1S,3Z,7S,11S,12S)-(+)-verticilla-3,8(19)-dien-7,12,20-triol 20-acetate (10). Acetylation of 9 and 10 yielded (1S,3Z,7S,11S,12S)-(+)-verticilla-3,8(19)-dien-7,12,20-triol 7,20-diacetate (11), while hydrolysis of 8 gave 7. The structures and stereochemistry of 7-11 were established by spectroscopic analyses, particularly by 1D and 2D NMR spectra and HRESIMS. The conformational preferences of 7-11 were studied by molecular mechanics modelling employing the Monte Carlo protocol followed by B3LYP/DGDZVP DFT calculation, thus supporting the observed ¹H NMR NOESY cross peaks.     

5α,8α-Epidioxysterols from the gorgonian Eunicella cavolini and the ascidian Trididemnum inarmatum: Isolation and evaluation of their antiproliferative activity

Three new (1, 4, 9) and nine previously reported (2, 3, 5-8, 10-12) 5α,8α-epidioxysterols were isolated from the organic extracts of the gorgonian Eunicella cavolini and the ascidian Trididemnum inarmatum. The structures and relative configurations of 1-12 were established on the basis of detailed NMR spectroscopic analyses and comparison with the literature. The growth inhibitory effects of 1-12 were evaluated against MCF-7 human breast cancer cells. Compound 1, bearing a cyclopropyl moiety in the side chain, exhibited the highest antiproliferative activity.