A Unique Population in a Unique Area: The Alcon Blue Butterfly and Its Specific Parasitoid in the Bialowieza Forest.

Caterpillars of the Alcon blue butterfly Phengaris alcon are initially endophytic and feed inside the flowerheads of Gentiana plants, but complete their development as social parasites in the nests of Myrmica ants, where they are fed by workers. Its specific and complicated ecological requirements make P. alcon a very local, threatened species, sensitive to environmental changes. We investigated an isolated and previously unknown population in an area of high nature value-the Bialowieza Forest (NE Poland). Using the mark-release-recapture method we estimated the seasonal number of adults at 1460 individuals, and their density (850/ha) was the highest among all populations using G. pneumonanthe studied so far. The site is also unique due to the presence of the specific parasitoid Ichneumon cf. eumerus, and parasitoids are considered the ultimate indicators of the biodiversity of Phengaris systems. Since 75.5% of P. alcon pupae were infested we could estimate the seasonal population of adult wasps at about 4500 individuals. The high abundance of both P. alcon and its parasitoid may be explained by favorable habitat characteristics, i.e., the strong presence of host plants and the high density of nests of Myrmica scabrinodis, which is the only local host ant of the butterfly.

Population Genetic Structure and Demography of the Critically Endangered Chequered Blue Butterfly (Scolitantides orion) in a Highly Isolated Part of Its Distribution Range.

Scolitantides orion is a butterfly species threatened in many European countries. In Poland, it survived in a single highly isolated area (Vistula River valley), which is an example of the dramatic decline in the population number. We studied the two largest remaining populations inhabiting opposite banks of the river. Mark-release-recapture studies showed that both populations were small, and they fluctuated in numbers, but adult individuals were twice as numerous on the western site. Genetic analyses were carried out using a mitochondrial (COI, ND5) and nuclear markers (Wgl, EF-1α, and microsatellite loci). We found out that genetic variation was low at both sites but higher in the smaller eastern population. This pattern is likely to be better explained by past distribution, when the butterfly, as a continental species used to be much more widespread in the east. However, the genetic differentiation between populations was low. This could suggest that the existing gene flow is facilitated by dominant regional wind direction, which may also contribute to a better genetic condition of the western population. Finally, a comparison of the obtained COI sequences with others available enabled us to reveal the phylogeographic pattern of the S. orion from different localities within its range.

Sexual differences in age-dependent survival and life span of adults in a natural butterfly population.

Adult survival and longevity in insects are key life-history traits, but their variation between sexes and individuals in natural populations is largely unexplored. Sexual divergence in senescence, the decline in survival with age is also poorly understood. Based on an intensive mark-recapture dataset of the butterfly Polyommatus daphnis, we aimed to assess whether adult survival is age-dependent, and to estimate life span distribution and abundance of males and females using Cormack-Jolly-Seber and Jolly-Seber models. Female survival slightly increased with date of emergence and slightly decreased with age, while male survival considerably declined with age. Mean life span of females (12.7 days) was ~50% higher than that of males (8.5 days), but two times higher if only the oldest 5% of each sex was considered (39 vs.19 days). Abundance of females (358 ± 14) and males (359 ± 11) was similar, but peak abundance of males preceded that of females by 11 days. Our results suggest that senescence is much more rapid in males than in females in this butterfly, which is in agreement with sexual selection theory. We also conclude that estimating life span distributions provides much more valuable information on the demography of natural populations than simply reporting the mean life span.

Patterns of host use by brood parasitic Maculinea butterflies across Europe.

The range of hosts exploited by a parasite is determined by several factors, including host availability, infectivity and exploitability. Each of these can be the target of natural selection on both host and parasite, which will determine the local outcome of interactions, and potentially lead to coevolution. However, geographical variation in host use and specificity has rarely been investigated. Maculinea (= Phengaris) butterflies are brood parasites of Myrmica ants that are patchily distributed across the Palæarctic and have been studied extensively in Europe. Here, we review the published records of ant host use by the European Maculinea species, as well as providing new host ant records for more than 100 sites across Europe. This comprehensive survey demonstrates that while all but one of the Myrmica species found on Maculinea sites have been recorded as hosts, the most common is often disproportionately highly exploited. Host sharing and host switching are both relatively common, but there is evidence of specialization at many sites, which varies among Maculinea species. We show that most Maculinea display the features expected for coevolution to occur in a geographic mosaic, which has probably allowed these rare butterflies to persist in Europe. This article is part of the theme issue 'The coevolutionary biology of brood parasitism: from mechanism to pattern'.

Cross-continental phylogeography of two Holarctic Nymphalid butterflies, Boloria eunomia and Boloria selene.

Pleistocene glaciations had significant effects on the distribution and evolution of species inhabiting the Holarctic region. Phylogeographic studies concerning the entire region are still rare. Here, we compared global phylogeographic patterns of one boreo-montane and one boreo-temperate butterflies with largely overlapping distribution ranges across the Northern Hemisphere, but with different levels of range fragmentation and food specialization. We reconstructed the global phylogeographic history of the boreo-montane specialist Boloria eunomia (n = 223) and of the boreo-temperate generalist Boloria selene (n = 106) based on mitochondrial and nuclear DNA markers, and with species distribution modelling (SDM). According to the genetic structures obtained, both species show a Siberian origin and considerable split among populations from Nearctic and Palaearctic regions. According to SDMs and molecular data, both butterflies could inhabit vast areas during the moderate glacials. In the case of B. selene, high haplotype diversity and low geographic structure suggest long-lasting interconnected gene flow among populations. A stronger geographic structuring between populations was identified in the specialist B. eunomia, presumably due to the less widespread, heterogeneously distributed food resources, associated with cooler and more humid climatic conditions. Populations of both species show opposite patterns across major parts of North America and in the case of B. eunomia also across Asia. Our data underline the relevance to cover entire distribution ranges to reconstruct the correct phylogeographic history of species.

Wolbachia infections mimic cryptic speciation in two parasitic butterfly species, Phengaris teleius and P. nausithous (Lepidoptera: Lycaenidae).

Deep mitochondrial divergence within species may result from cryptic speciation, from phylogeographic isolation or from endosymbiotic bacteria like Wolbachia that manipulate host reproduction. Phengaris butterflies are social parasites that spend most of their life in close relationship with ants. Previously, cryptic speciation has been hypothesised for two Phengaris species based on divergent mtDNA sequences. Since Phengaris species are highly endangered, the existence of cryptic species would have drastic consequences for conservation and management. We tested for cryptic speciation and alternative scenarios in P. teleius and P. nausithous based on a comprehensive sample across their Palaearctic ranges using COI gene sequences, nuclear microsatellites and tests for Wolbachia. In both species a deep mitochondrial split occurring 0.65-1.97 myrs ago was observed that did not correspond with microsatellite data but was concordant with Wolbachia infection. Haplotypes previously attributed to cryptic species were part of the Wolbachia-infected clades. In both species remaining phylogeographic structure was largely consistent between mitochondrial and nuclear genomes. In P. teleius several mitochondrial and nuclear groups were observed in East Asia while a single haplogroup and nuclear cluster prevailed across continental Eurasia. Neutrality tests suggested rapid demographic expansion into that area. In contrast, P. nausithous had several mitochondrial and nuclear groups in Europe, suggesting a complex phylogeographic history in the western part of the species range. We conclude that deep intraspecific divergences found in DNA barcode studies do not necessarily need to represent cryptic speciation but instead can be due to both infection by Wolbachia and phylogeographic structure.

Contrasting genetic structure of rear edge and continuous range populations of a parasitic butterfly infected by Wolbachia.

BACKGROUND: Climatic oscillations are among the long-term factors shaping the molecular features of animals and plants and it is generally supposed that the rear edges (i.e., the low-latitude limits of distribution of any given specialised species) situated closer to glacial refugia are vital long-term stores of genetic diversity. In the present study, we compared the genetic structure of several populations of an endangered and obligate myrmecophilous butterfly (Maculinea arion) from two distinct and geographically distant parts of its European distribution (i.e., Italy and Poland), which fully represent the ecological and morphological variation occurring across the continent. RESULTS: We sequenced the COI mitochondrial DNA gene (the 'barcoding gene') and the EF-1α nuclear gene and found substantial genetic differentiation among M. arion Italian populations in both markers. Eleven mtDNA haplotypes were present in Italy. In contrast, almost no mtDNA polymorphisms was found in the Polish M. arion populations, where genetic differentiation at the nuclear gene was low to moderate. Interestingly, the within-population diversity levels in the EF-1α gene observed in Italy and in Poland were comparable. The genetic data did not support any subspecies divisions or any ecological specialisations. All of the populations studied were infected with a single strain of Wolbachia and our screening suggested 100% prevalence of the bacterium. CONCLUSIONS: Differences in the genetic structure of M. arion observed in Italy and in Poland may be explained by the rear edge theory. Although we were not able to pinpoint any specific evolutionarily significant units, we suggest that the Italian peninsula should be considered as a region of special conservation concern and one that is important for maintaining the genetic diversity of M. arion in Europe. The observed pattern of mtDNA differentiation among the populations could not be explained by an endosymbiotic infection.

Mimetic host shifts in an endangered social parasite of ants.

An emerging problem in conservation is whether listed morpho-species with broad distributions, yet specialized lifestyles, consist of more than one cryptic species or functionally distinct forms that have different ecological requirements. We describe extreme regional divergence within an iconic endangered butterfly, whose socially parasitic young stages use non-visual, non-tactile cues to infiltrate and supplant the brood in ant societies. Although indistinguishable morphologically or when using current mitochondrial and nuclear sequence-, or microsatellite data, Maculinea rebeli from Spain and southeast Poland exploit different Myrmica ant species and experience 100 per cent mortality with each other's hosts. This reflects major differences in the hydrocarbons synthesized from each region by the larvae, which so closely mimic the recognition profiles of their respective hosts that nurse ants afford each parasite a social status above that of their own kin larvae. The two host ants occupy separate niches within grassland; thus, conservation management must differ in each region. Similar cryptic differentiation may be common, yet equally hard to detect, among the approximately 10 000 unstudied morpho-species of social parasite that are estimated to exist, many of which are Red Data Book listed.