Morphological and chemical analysis of male scent organs in the butterfly genus Pyrgus (Lepidoptera: Hesperiidae)

Chemical communication in the family Hesperiidae (Lepidoptera) is practically unstudied, even though this group includes approximately 4,000 species and represents a fifth of the world's butterfly fauna. We present the first comparative morphological and chemical analysis of scent organs for nine species in the genus Pyrgus, the most species-rich hesperiid genus in the Palearctic region. Our results show that the morphology of the two main male scent organs - the costal fold and the tibial tufts - does not differ between species. The chemical analyses detected a total of 125 different compounds exclusively present in these organs. We document great interspecific differences and much narrower intraspecific variability in the chemical profiles. The dynamics of chemical versus genetic distances indicate two different phases: a faster (but more variable) initial chemical divergence at lower genetic divergences (probably related to speciation) and a slower but more constant differentiation (drift). As a result most species can be identified based on their chemical profiles, except for a closely related species pair (P. malvae/P. malvoides) for which hybridisation is common in the contact zone. Our results suggest that the Hesperiidae is a group with great potential for the study of chemical communication that deserves further attention. © 2014 Gesellschaft für Biologische Systematik.Funding for this research was provided by Universitat Autònoma de Barcelona (project EME2007-33 to J.L.H.-R., R.B. and R.V.) and by the Spanish Ministerio de Ciencia e Innovación (project CGL2010-21226/BOS to J.L.H.-R. and R.V. and project CGL2004-04680-c10-08/BOS to M.L.M.).Peer Reviewe

Genomics of extreme ecological specialists: multiple convergent evolution but no genetic divergence between ecotypes of Maculinea alcon butterflies.

Biotic interactions are often acknowledged as catalysers of genetic divergence and eventual explanation of processes driving species richness. We address the question, whether extreme ecological specialization is always associated with lineage sorting, by analysing polymorphisms in morphologically similar ecotypes of the myrmecophilous butterfly Maculinea alcon. The ecotypes occur in either hygric or xeric habitats, use different larval host plants and ant species, but no significant distinctive molecular traits have been revealed so far. We apply genome-wide RAD-sequencing to specimens originating from both habitats across Europe in order to get a view of the potential evolutionary processes at work. Our results confirm that genetic variation is mainly structured geographically but not ecologically - specimens from close localities are more related to each other than populations of each ecotype from distant localities. However, we found two loci for which the association with xeric versus hygric habitats is supported by segregating alleles, suggesting convergent evolution of habitat preference. Thus, ecological divergence between the forms probably does not represent an early stage of speciation, but may result from independent recurring adaptations involving few genes. We discuss the implications of these results for conservation and suggest preserving biotic interactions and main genetic clusters

The decline of the charismatic Parnassius mnemosyne (L.) (Lepidoptera: Papilionidae) in a central italy national park: A call for urgent actions

Here we report the strong decline of a population of the endangered species Parnassius mnemosyne (Linnaeus, 1758) (Lepidoptera: Papilionidae) in the National Park of Foreste Casentinesi, Monte Falterona and Campigna (Italy). We compared historical presence data (before 1969) with current data from two years of monitoring (2018–2019) and provided evidence of a drastic reduction in the number of sites inhabited by the species. A preliminary assessment suggested that the population of P. mmemosyne occurring in the Park is limited to a few individuals. We argue that the population of this iconic and charismatic butterfly is at the verge of extinction in this National Park, probably because of a combination of habitat loss (i.e. decrease in size and number of areas of open grassland on the mountain belt) and climatic changes. Being one of the few populations of P. mnemosyne in the Northern Apennines, the implementation of protection measures is a high conservation priority. Several other butterfly species on the Italian mountains that are facing the same survival challenges would also benefit from the establishment of conservation actions aimed at improving habitat quality for P. mnemosyne. Present article is meant to call for action researchers, stakeholders, and especially decision-makers in order to increase the efforts to upturn the evident decline in abundance of this population

Simple and informative: applying a basic Anthophila monitoring scheme in a simplified insular ecosystem

The decline of pollinators and the consequent decay of pollination services call for the establishment of monitoring schemes for several groups of pollinators. For Anthophila (Hymenoptera), the design of monitoring schemes is still under development. The main difficulties lie in combining a reliable but field-feasible taxonomic identification with the collection of informative data about the consistency and functional role of pollinator populations. Here we report on the application of the Italian monitoring scheme for pollinators recently defined by ISPRA and the University of Turin in agreement with the European Pollinators Monitoring Scheme on the small island of Giannutri (Tuscany), a simplified insular ecosystem with a virtually unknown pollinator community. This island has recently experienced a drastic change in its bee community, as since 2018 honey bee (Apis mellifera L.) hives are regularly moved every year to the island for breeding purposes. In the spring 2021 we established six 250 m long fixed transects and performed a total of 48 surveys (8 for each transect), recording more than 2300 observations of 9 Anthophila bee taxa and the flowers they visited. By using generalised additive mixed models, we showed that the monitoring protocol has a good potential for monitoring Anthophila, as we could verify several expected relationships between Anthophila abundance and abiotic factors (season, hour of the day, distance from the apiary) and biotic factors (abundance of flower resources). More importantly, we verified that A. mellifera represents by far the most frequent Anthophila taxon. Our data do not show evidence for spatial partition between A. mellifera and the other most frequent taxa (Bombus terrestris L. and Anthophora spp.). The visit network based on transect observations also showed that these taxa largely overlapped in terms of visits to flower resources. Overall, our data showed that the monitoring protocol allows gathering informative data about Anthophila taxa abundance, interactions and flower-visits. Moreover, the spatial and flower-visit overlap suggest potential for competition between honey bees and wild pollinators, with a potential consequent resource depletion for the latter. While this hypothesis could only be assessed by a long-term monitoring and ad hoc honey bee removal experiments, our data show that this basic monitoring protocol produces rapid and valuable information about Anthophila community and dynamics

The decline of the charismatic Parnassius mnemosyne (L.) (Lepidoptera: Papilionidae) in a Central Italy national park: a call for urgent actions

Here we report the strong decline of a population of the endangered species Parnassius mnemosyne (Linnaeus, 1758) (Lepidoptera: Papilionidae) in the National Park of Foreste Casentinesi, Monte Falterona and Campigna (Italy). We compared historical presence data (before 1969) with current data from two years of monitoring (2018–2019) and provided evidence of a drastic reduction in the number of sites inhabited by the species. A preliminary assessment suggested that the population of P. mmemosyne occurring in the Park is limited to a few individuals. We argue that the population of this iconic and charismatic butterfly is at the verge of extinction in this National Park, probably because of a combination of habitat loss (i.e. decrease in size and number of areas of open grassland on the mountain belt) and climatic changes. Being one of the few populations of P. mnemosyne in the Northern Apennines, the implementation of protection measures is a high conservation priority. Several other butterfly species on the Italian mountains that are facing the same survival challenges would also benefit from the establishment of conservation actions aimed at improving habitat quality for P. mnemosyne. Present article is meant to call for action researchers, stakeholders, and especially decision-makers in order to increase the efforts to upturn the evident decline in abundance of this population

Enhancement of the Biological and Mechanical Performances of Sintered Hydroxyapatite by Multiple Ions Doping

In the present work, hydroxyapatite (HA) nanoparticles doped with Mg2+, Sr2+, and Zn2+ ions are developed by wet neutralization method and then sintered at 1,250°C to obtain bulk consolidated materials. Physicochemical and microstructural analyses show that the presence of doping ions in the HA structure induced the formation of βTCP as secondary phase, during the sintering process, and we found that this effect is depending on the stability of the various doping ions in the hydroxyapatite lattice itself. We also found that the formation of βTCP as secondary phase, in turn, confines the grain growth of HA induced by the high-temperature sintering process, thus leading to a strong increase of the flexural strength of the bulk materials, according to Hall-Petch-like law. Furthermore, we found that the doping ions enter also in the structure of the βTCP phase; besides the grain growth confinement, also the solubility and ion release ability of the final materials were enhanced. In addition to ameliorate the mechanical performance, the described phenomena also activate multiple biofunctionalities: (i) ability to upregulate various genes involved in the osteogenesis, as obtained by human adipose stem cells culture and evaluated by array technology; (ii) enhanced resistance to the adhesion and proliferation of Gram+ and Gram– bacterial strains. Hence, our results open a perspective for the use of sintered multiple ion-doped HA to develop ceramic biodevices, such as plates, screws, or other osteosynthesis media, with enhanced strength, osteointegrability, and the ability to prevent post-surgical infections

The isolated Erebia pandrose Apennine population is genetically unique and endangered by climate change

Climate change is causing shifts in the distribution of many species and populations inhabiting mountain tops are particularly vulnerable to these threats because they are constrained in altitudinal shifts. Apennines are a relatively narrow and low mountain chain located in Southern Europe, which hosts many isolated populations of mountain species. The butterfly Erebia pandrose was recorded for the last time in the Apennines in 1977, on the top of a single massif (Monti della Laga). We confirmed the presence of a small, isolated population of E. pandrose in the Apennines, at a distance of more than 400 km to any other known populations. Then, we examined the cytochrome c oxidase subunit 1 mitochondrial DNA marker of this species across the Palaearctic area and estimated the potential decline over the Alps and the Apennines due to future climatic changes. The Apennine population represents an endemic lineage characterised by eight mutations over the 658 bp analysed (1.2%). In the Alps and Apennines, this species has shifted uphill more than 3 m per year since the end of the 19th century and more than 22 m per year since 1995. Species distribution models suggested that these mountain populations will experience a generalised loss of climatic suitability, which, according to our projections, could lead to the extinction of the Apennine population in a few decades. Erebia pandrose has the potential to become a flagship species for advertising the risk of losing unique fractions of genetic diversity for mountain species

Unrelated Helpers in a Primitively Eusocial Wasp: Is Helping Tailored Towards Direct Fitness?

The paper wasp Polistes dominulus is unique among the social insects in that nearly one-third of co-foundresses are completely unrelated to the dominant individual whose offspring they help to rear and yet reproductive skew is high. These unrelated subordinates stand to gain direct fitness through nest inheritance, raising the question of whether their behaviour is adaptively tailored towards maximizing inheritance prospects. Unusually, in this species, a wealth of theory and empirical data allows us to predict how unrelated subordinates should behave. Based on these predictions, here we compare helping in subordinates that are unrelated or related to the dominant wasp across an extensive range of field-based behavioural contexts. We find no differences in foraging effort, defense behaviour, aggression or inheritance rank between unrelated helpers and their related counterparts. Our study provides no evidence, across a number of behavioural scenarios, that the behaviour of unrelated subordinates is adaptively modified to promote direct fitness interests

Butterfly-parasitoid-hostplant interactions in Western Palaearctic Hesperiidae: a DNA barcoding reference library

The study of ecological interactions between plants, phytophagous insects and their natural enemies is an essential but challenging component for understanding ecosystem dynamics. Molecular methods such as DNA barcoding can help elucidate these interactions. In this study, we employed DNA barcoding to establish hostplant and parasitoid interactions with hesperiid butterflies, using a complete reference library for Hesperiidae of continental Europe and north-western Africa (53 species, 100% of those recorded) based on 2934 sequences from 38 countries. A total of 233 hostplant and parasitoid interactions are presented, some recovered by DNA barcoding larval remains or parasitoid cocoons. Combining DNA barcode results with other lines of evidence allowed 94% species-level identification for Hesperiidae, but success was lower for parasitoids, in part due to unresolved taxonomy. Potential cases of cryptic diversity, both in Hesperiidae and Microgastrinae, are discussed. We briefly analyse the resulting interaction networks. Future DNA barcoding initiatives in this region should focus attention on north-western Africa and on parasitoids, because in these cases barcode reference libraries and taxonomy are less well developed.Support for this research was provided by the Spanish National Research Council (CSIC) with a JAE-Intro fellowship for the introduction to research to ETD (reference numbers JAEINT_20_00248 and JAEINT20_EX_0638) and by projects PID2019-107078GB-I00/MCIN/AEI/10.13039/501100011033 and 2017-SGR-991 (Generalitat de Catalunya) to RV, and PID2020-117739GA-I00/MCIN/AEI/10.13039/501100011033 to GT. We thank the Rachadaphiseksomphot Fund, Graduate School, Chulalongkorn University, for the award of a Senior Postdoctoral Fellowship to DLJQ. Further support for this research was provided by the Academy of Finland (Academy Research Fellow, decision no. 328895) to VD. PDNH acknowledges support from Genome Canada through Ontario Genomics. BV has been funded by the CERCA Programme of the Generalitat de Catalunya and by the Grant RYC-22243-2017, whose PI is Josep Sardanyés. SV was supported by the Spanish Ministry of Economy and Competitiveness, grant PID2020-117822GB-I00 MINEICO/AEI/ FEDER and the European Union.INTRODUCTION MATERIAL AND METHODS RESULTS DISCUSSION CONCLUSION SUPPORTING INFORMATION ACKNOWLEDGEMENTS DATA AVAILABILITY REFERENCES Supplementary dat