Interspecific relationships in co‐occurring populations of social parasites and their host ants

Myrmica ant colonies host numerous insect species, including the larvae of Maculinea butterflies and Microdon myrmicae hoverflies. Little is known about the interspecific relationships among these social parasites and their host ants occurring in sympatric populations. We investigated communities of social parasites to assess the strategies allowing them to share the same pool of resources (i.e. Myrmica colonies). The present study was carried out at five sites inhabited by different social parasite communities, each comprising varying proportions of Maculinea teleius, Maculinea nausithous, Maculinea alcon, and Microdon myrmicae. We investigated their spatial distributions, host segregation, the degree of chemical similarity between social parasites and hosts, and temporal overlaps in colony resource exploitation. Spatial segregation among social parasites was found in two populations and it arises from microhabitat preferences and biological interactions. Local conditions can drive selection on one social parasite to use a Myrmica host species that is not exploited by other social parasites. Myrmica scabrinodis and Myrmica rubra nests infested by larvae of two social parasite species were found and the most common co‐occurrence was between Ma. teleius and Mi. myrmicae. The successful coexistence of these two species derives from their exploitation of the host colony resources at different times of the year. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 699–709.     

Local populations of endangered Maculinea (Phengaris) butterflies are flood resistant

Semi-natural wet meadows are threatened by drainage, the abandonment of traditional management and climate change. The large blue butterflies Maculinea teleius and M. nausithous are flagship species associated with wet meadows and are the targets of many conservation programmes. However, there is little knowledge on the impact of natural catastrophes, such as floods, on the persistence of these butterflies. In our study we tested how a flood that resulted in the temporary inundation of meadows affected populations of M. teleius and M. nausithous. Studies were conducted in two consecutive seasons of 2009 (with ‘normal’ weather) and 2010 (with extreme rainfall and a consequent flood in May) in a wet meadow complex located in the Vistula River valley in southern Poland. In both years the abundance of adults was estimated for each local habitat patch (n = 55) within sympatric metapopulations of both species. Additionally, in June 2010, i.e. directly after the flood and shortly before the flight period, a total of 754 Myrmica ant nests in 10 habitat patches (6 inundated vs. 4 not inundated) were checked for the presence of Maculinea larvae and pupae. We found no impact of inundation on year-to-year changes in adult population sizes. The probability of occurrence of Maculinea larvae and pupae in ant nests was higher in temporarily inundated meadows. Our results indicate that temporary inundation occurring after long-term downpours does not negatively affect the investigated species even during the larval period in ant nests at ground level. This provides an argument against drainage works in wet meadows with Maculinea butterflies.     

All natural habitat edges matter equally for endangered <Emphasis Type="Italic">Maculinea</Emphasis> butterflies

An obvious consequence of habitat fragmentation is an increasing role of habitat edges for species survival. Recently it has been suggested that the endangered butterfly Maculinea nausithous prefers forested edges of its meadow habitats. However, the prevalence of forests in the study area used for this analysis makes it impossible to distinguish whether the effect detected is a genuine preference for forest edges or a preference for any natural patch edges as opposed to patch interiors. We investigated habitat selection by Maculinea nausithous and Maculinea teleius occurring sympatrically at five habitat patches surrounded by mosaic landscape. Butterfly capture positions were marked with GPS and subsequently analysed with GIS software. Both species avoided the interiors of their patches and concentrated in the edge zone, but these preferences were visible only at three larger patches exceeding 1 ha in area. Among different types of edges those bordering densely built-up areas were avoided, whereas all natural edges (adjacent to forests, reeds or grasslands) were similarly used. We hypothesise that preferences towards natural patch edges, regardless of their type, can be explained by the spatial interactions between Maculinea butterflies and Myrmica ants they parasitise. Patch surroundings constitute refuge space for the ants, and hence their densities may be expected to be higher near patch edges. Our findings indicate the importance of patch surroundings for the persistence of Maculinea populations. Regretfully, current legal framework makes it difficult to protect patch surroundings, where neither priority species nor their habitats occur.     

Microsatellite markers for the large blue butterflies Maculinea nausithous and Maculinea alcon (Lepidoptera: Lycaenidae) and their amplification in other Maculinea species

We developed microsatellite markers for Maculinea nausithous and Maculinea alcon, two of five species of endangered large blue butterflies found in Europe. Two separate microsatellite libraries were constructed. Eleven markers were developed for M. nausithous and one for M. alcon. The primers were tested on both species as well as on the three other European Maculinea species. The number of alleles per locus ranged from two to 14. These markers will be useful tools for population genetic studies of Maculinea species.     

Polymorphic growth in larvae of Maculinea butterflies, as an example of biennialism in myrmecophilous insects

The presence of annual and biennial individuals within the same population has been recently demonstrated in the myrmecophilous butterflies Maculinea rebeli and Maculinea alcon, which present a cuckoo strategy inside Myrmica nests, and Maculinea arion which is a predatory species. Here, we present field and laboratory data on polymorphic larval growth in two other predatory species of Maculinea: M. teleius and M. nausithous. Body mass distributions of pre-pupation larvae were bimodal in both species. These results point to the existence of larvae that develop in 1 or 2 years. We also showed that the probability of pupation depended on larval body mass. In the case of M. teleius, the critical body mass at which larvae have a 50% probability of pupation is about 80 mg. We suggest that polymorphism in Maculinea may have evolved as an adaptation to life in ant nests, a habitat which protects them from predators and provides food. However, the quality of this resource is highly variable and unpredictable. According to the bet-hedging hypothesis, if the habitat is unpredictable, females should have an advantage by producing more variable offspring. In the case of Maculinea butterflies, this may involve maintaining larvae that develop in 1 or 2 years.     

Species-specific distribution of two sympatric Maculinea butterflies across different meadow edges

An important consequence of habitat fragmentation is the increase of edge habitats. Environmental factors in the edges are different from those in the interiors, which causes changes in the distribution of plant and animal species. We aimed to study how edges affect the distribution of two butterfly species within meadow fragments. We therefore investigated the effect of distance from edge and edge type (road edge versus tree edge) on two sympatric large blue species (Maculinea teleius and M. nausithous). Our results showed that edge type had contrasting effects on the two species. M. teleius favoured both interiors and road edges, while M. nausithous preferred the tree edges. In the case of the latter species a strong positive edge effect was also found. This kind of within-habitat niche segregation is probably related to the different microenvironmental conditions at the edges. Foodplant density did not seem to limit the distribution of these species. Our results suggest that interiors of meadows are important for M. teleius, while tree edges maintain the habitats of the regionally rarer butterfly, M. nausithous.     

Phylogeny of Maculinea blues (Lepidoptera: Lycaenidae) based on morphological and ecological characters: evolution of parasitic myrmecophily

A phylogeny of blue butterflies of the genus Maculinea and related genera (Lycaenidae) is proposed, based on 91 morphological and ecological characters. The resulting tree shows that: (1) Phengaris is a derived group nested within Maculinea; (2) the Maculinea‐Phengaris clade is probably nested within Glaucopsyche; (3) there are three well supported groups within the Maculinea‐Phengaris clade: (alcon group ((teleius group) (arion‐Phengaris group))). Some species (M. alcon, M. arionides) appear to be non‐monophyletic and require reclassification. The two alternative strategies of parasitic myrmecophily in the Maculinea‐Phengaris clade, viz., “predatory” and “cuckoo”, seem to be derived characters of the alcon group, and of the teleius and arion‐Phengaris groups, respectively. The common ancestor of Maculinea used dorsal nectary organ secretions for ant attraction, while this trait was reduced in the ancestor of the alcon group and in M. nausithous (of the teleius group). The three recent Maculinea lineages utilize taxonomically diverse host plants, the asterid families Gentianaceae (alcon and arion‐Phengaris groups), Lamiaceae (arion‐Phengaris group), Campanulaceae (arion‐Phengaris group), and the rosid family Rosaceae (teleius group).     

Population ecology of the endangered butterflies Maculinea teleius and M. nausithous and the implications for conservation

Butterflies of the genus Maculinea are highly endangered in Europe. The cuckoo species M. rebeli has been thoroughly investigated through both empirical and modelling studies, but less is known about the population ecology of predatory Maculinea. We present the findings of a 2-year research study on sympatric populations of two endangered butterflies: Maculinea teleius and M. nausithous in the Kraków region, southern Poland. The study comprised mark–release–recapture sampling and laboratory rearing of butterflies from larvae collected in the field. For both species the sex ratio was slightly, but consistently, female-biased and there was little year-to-year change in the seasonal population sizes. Daily numbers showed greater variation between the 2 years of the study due to the differences in daily survival rate. The average life span of laboratory-raised butterflies kept in ideal conditions was more than 6 days, compared to only 2–3 days in the field. The recruitment of both males and females consistently followed a bimodal pattern. A small proportion of individuals (maximum 25%) changed sites, in spite of relatively short distances of ca. 100 m separating them. The results indicate that populations of both species are typically stable within their sites, possibly due to larval polymorphism, but there is little inter-site mobility and thus landscape corridors seem necessary to enhance metapopulation viability. A further problem to be considered in the conservation of Maculinea butterflies is the fact that their very short life span in relation to flight-period length reduces the effective population size.     

Comparison of genetic population structure of the large blue butterflies Maculinea nausithous and M. teleius

We investigated the genetic population structure of two rare myrmecophilous lycaenid butterflies, Maculinea nausithous and M. teleius, which often live sympatrically and have similar biology. In Europe, both species occur in highly fragmented populations and are vulnerable to local extinction. The proportion of variable allozyme loci, average heterozygosity and genetic differentiation among populations was higher in M. nausithous than in sympatrically living M. teleius populations. We hypothesise that the differences in heterozygosity are mainly due to the known higher efficiency of typical host ant nests in rearing M. nausithous pupae compared to M. teleius pupae. This implies a larger probability of larval survival in M. nausithous, which buffers populations against environmental and demographic stochasticity. In contrast, the lower carrying capacity of ant nests in rearing M. teleius pupae requires higher nest-densities and makes M. teleius populations more prone to losing genetic variation through drift if this condition is not fulfilled. The single investigated Russian population of M. teleius showed much higher levels of heterozygosity than any of the Polish populations, suggesting a more viable and still intact metapopulation structure.     

<Emphasis Type="Italic">Myrmica</Emphasis> host-ants limit the density of the ant-predatory large blue <Emphasis Type="Italic">Maculinea nausithous</Emphasis>

Butterflies of the highly endangered genus Maculinea are parasites of red Myrmica ants. Prior to the adoption by Myrmica worker ants Maculinea caterpillars feed on a specific host plant. This field study aims to answer the question whether the density and distribution of the host plant Sanguisorba officinalis or the density of the host ant M. rubra limit the density of M. nausithous egg, larval and adult stage. We found that the density of M. nausithous egg stage and adult stage increased with the density of the host ant. The density of M. nausithous caterpillars was not associated with ant density or plant density. This study suggests that the density of M. nausithous is limited by the density of the host ant M. rubra. We conclude that habitat management for M. nausithous should focus on the maintenance of habitats that hold both resources, but that enable high densities of M. rubra. In addition, it is discussed why high densities of host ants might be more important in predatory than in cuckoo-feeding Maculinea.     

Choice behaviour of Myrmica rubra workers between ant larvae and larvae of their Phengaris (Maculinea) nausithous nest parasites

Larvae of Phengaris (Maculinea) butterflies are adopted by Myrmica workers and are obligate myrmecophiles. Brood recognition by Myrmica rubra workers was tested for concolonial larvae (M. rubra) versus allocolonial larvae (M. rubra and P. nausithous) to assay the mimetic efficiency of P. nausithous. In addition, we tested M. rubra ant colonies from different populations with and without the presence of Phengaris, to test for potential local adaptation in adoption behaviour. We show that M. rubra can distinguish between nest-mate and foreign larvae as well as between P. nausithous and their own larvae. Workers from the allopatric population inspected and rejected more P. nausithous larvae than workers from the sympatric population. This might reflect a local host adaptation in which the social parasite more efficiently mimes its sympatric host ants than allopatric ones.     

Development of parasitic <Emphasis Type="Italic">Maculinea teleius</Emphasis> (Lepidoptera,Lycaenidae) larvae in laboratory nests of four <Emphasis Type="Italic">Myrmica</Emphasis> ant host species

Maculinea butterflies are social parasites of Myrmica ants. Methods to study the strength of host ant specificity in the Maculinea–Myrmica association include research on chemical and acoustic mimicry as well as experiments on ant adoption and rearing behaviour of Maculinea larvae. Here we present results of laboratory experiments on adoption, survival, development and integration of M. teleius larvae within the nests of different Myrmica host species, with the objective of quantifying the degree of specialization of this Maculinea species. In the laboratory, a total of 94 nests of four Myrmica species: M. scabrinodis, M. rubra, M. ruginodis and M. rugulosa were used. Nests of M. rubra and M. rugulosa adopted M. teleius larvae more readily and quickly than M. ruginodis colonies. No significant differences were found in the survival rates of M. teleius larvae reared by different ant species. Early larval growth of M. teleius larvae differed slightly among nests of four Myrmica host species. Larvae reared by colonies of M. rugulosa which were the heaviest at the beginning of larval development had the lowest mean larval body mass after 18 weeks compared to those reared by other Myrmica species. None of the M. teleius larvae was carried by M. scabrinodis or M. rubra workers after ant nests were destroyed, which suggests a lack of integration with host colonies. Results indicate that Myrmica species coming from the same site differ in their ability to adopt and rear M. teleius larvae but there was no obvious adaptation of this butterfly species to one of the host ant species. This may explain why, under natural conditions, all four ants can be used as hosts of this butterfly species. Slight advantages of particular Myrmica species as hosts at certain points in butterfly larval development can be explained by the ant species biology and colony structure rather than by specialization of M. teleius.     

The importance of resource distribution: spatial co-occurrence of host plants and host ants coincides with increased egg densities of the Dusky Large Blue <Emphasis Type="Italic">Maculinea nausithous</Emphasis> (Lepidoptera: Lycaenidae)

The occurrence of the Dusky Large Blue Butterfly (Maculinea nausithous) critically depends on the availability of two key resources: the Great Burnet (Sanguisorba officinalis) as primary nectar source for adults, for egg laying and early larval development, and the host ant Myrmica rubra as the food of late instar larvae. Thus, their distributions are key parameters shaping habitat suitability, and we expected that overlapping of both resources would have a strong impact on the size of local M. nausithous populations. Their egg density may be affected (a) by the fraction of host plants per site located within My. rubra activity ranges at the patch scale, or (b) by the availability of host plants with host ants in close range at the local scale, due to the potential ability of butterfly females to detect their host ants. To test the above hypothesis, we recorded spatial distribution patterns of host plants and host ants on 29 study sites in south-western Germany and related them to egg density data of M. nausithous. We found a positive relationship between co-occurence of host plant and host ant and M. nausithous egg density at the patch scale, whereas no correlation was found at the local scale. Thus, focal populations are strongly limited by the abundance of host plants, covered with My. rubra activity, as ant-mediated oviposition could not be proved. Our results underline the importance of resource distribution; the understanding of its impacts may provide useful insights into how M. nausithous habitats can be managed in order increase their carrying capacity.     

Tetracosane on the cuticle of the parasitic butterfly Phengaris (Maculinea) nausithous triggers the first contact in the adoption process by Myrmica rubra foragers

Phengaris (Maculinea) butterflies are social parasites of Myrmica ant colonies. Larvae of the parasite are adopted by the ant workers into the colonies. Apparently, chemical signals are used by Phengaris nausithous Bergsträsser larvae to mimic those of the host brood to be recognized by the ants. In the present study, chemical extracts of ant brood and butterfly larvae using four different solvents are tested in behavioural choice assays in search of compounds involved in the adoption process. Tetracosane is the main shared compound in all brood extracts of Myrmica rubra L. and in all larvae of P. nausithous. The attractiveness of tetracosane for M. rubra workers is confirmed by testing synthetic tetracosane in behavioural choice assays, suggesting that the adoption ritual may be initiated by tetracosane.     

A mowing experiment to evaluate the influence of management on the activity of host ants of <Emphasis Type="Italic">Maculinea</Emphasis> butterflies

Maculinea butterflies obligatory parasitize certain species of Myrmica ants. Thus, the presence of the host ant species is a limiting factor for the survival of a Maculinea population. Here, we analyse the influence of vegetation structure and ground temperature on ant diversity and abundance on Maculinea habitats, with the final aim of identifying the environmental variables determining patterns of variation in species composition in order to recommend a mowing regime that will promote our three target species: Maculinea teleius, M. nausithous and M. alcon. Experimental plots with different mowing regimes were established at eight sites in South-Eastern Germany, a region which still contains a number of relatively large, stable populations of these threatened butterfly species. Among the seven different ant species recorded, four belong to the genus Myrmica (M. scabrinodis, M. rubra, M. ruginodis and M. vandeli). Among these, M. scabrinodis results most abundant at all sites. In a CCA analysis of environmental variables recorded at the studied plots, ant species diversity appears largely determined by litter cover, mean temperature, and mean grass cover. Mowing once a year, in the second half of September, after the larvae have left their host plants, enhances the abundance of Myrmica ants in the meadows, and would be the best management compromise for all three species.     

The ecology of Myrmica ants in relation to the conservation of Maculinea butterflies

Maculinea butterflies in Europe, and probably most of Asia, are host specific social parasites of various species of Myrmica ants. The latest summary of field data showing the pattern of host specificity by Maculinea is presented. Myrmica ants have been well studied in the laboratory but much less is known about the ecology of their natural populations. While the former is important in understanding the adaptive evolution of Maculinea larval behaviours, the latter is of more practical importance to conservationists charged with the protection of specific populations of Maculinea. The current knowledge of habitat partition, colony growth and colony reproduction within communities of Myrmica ants is summarized in relation to the ecology of Maculinea species. Concepts used in current population simulation models are explained. A key concept is the idea that community structure (both number of species and size and abundance of nests) is controlled by the quantity and quality of suitable nest sites. Some advice is given to conservationists who might need to manipulate Myrmica ant populations in order to maintain a robust population of a Maculinea species. © Rapid Science Ltd. 1998     

Patterns of host ant use by sympatric populations of <Emphasis Type="Italic">Maculinea alcon</Emphasis> and <Emphasis Type="Italic">M. ‘rebeli’</Emphasis> in the Carpathian Basin

Maculinea butterflies show social parasitism via obligatory myrmecophily as their larvae are adopted and raised to pupation by Myrmica ants. Suitable hosts differ for different Maculinea species, and host ant specificity can further differ at the population-level. Although early studies suggested single ant species as main hosts for each Maculinea species, it has recently become clear that their host ant specificity is more complex. Maculinea alcon and Maculinea ‘rebeli’ have variously been separated according to adult and larval morphology, phenology, and their use of different ecosystems, including host plant and host ant species. However, recent genetic evidence has questioned their separation as good species. Here we compare the use of host ants by M. alcon and M. ‘rebeli’ at the regional scale in NE-Hungary and Transylvania (Romania), where molecular studies have found no species-level separation between the two forms. We opened 778 nests of Myrmica ants and searched for Maculinea specimens (larvae, pupae and exuviae) shortly before imago emergence from the nest in seven M. alcon sites, six M. ‘rebeli’- sites and one site where both M. alcon and M. ‘rebeli’ are syntopic. In all, Maculinea caterpillars were found in the nests of seven different ant species (M. alcon was recorded mainly with Myrmica scabrinodis and occasionally with M. salina and M. vandeli; M. ‘rebeli’ used mainly M. scabrinodis, M. sabuleti and M. schencki and occasionally M. lonae and M. specioides). Myrmica scabrinodis was found to be a general host of both M. alcon and M. ‘rebeli’, which is the first record for a common host ant of these two closely related butterflies within the same region. However there were also differences in host ant use patterns between the sites occupied by the two Maculinea taxa, which reflect differences in Myrmica communities between the two types of habitat. Possible explanations for the similar but not identical host use patterns of M. alcon and M. ‘rebeli’, and their relevance for the question of whether they are separate species are discussed. Received 27 November 2007; revised 28 May 2008; accepted 11 June 2008.     

Movements and flight morphology in the endangered Large Blue butterflies

Movements and flight morphology of the endangered Large Blue butterflies Phengaris teleius and P. nausithous in southern Poland were studied with mark-release-recapture surveys and GIS analyses. Most individuals moved relatively small distances (<40 metres) within their habitat patches. Distances covered by both species were positively related to season progression and wing length, and negatively related to body mass. P. teleius movement distances slightly exceeded those of P. nausithous. In addition, females moved longer distances than males, although the difference was significant only in P. teleius. Morphological traits appeared to be good indicators of the inter-specific and inter-sexual differences in mobility. While P. teleius individuals were heavier than P. nausithous ones, they had considerably longer wings, which may explain longer movements in the former species. Similarly, females were heavier than males in both species, but they invested more in wing size, which is likely to compensate for the negative impact of body mass on movement distances. Our results indicate that combination of GIS analysis of movement distances recorded with mark-release-recapture methods and morphometric measurements taken in field during non-lethal handling of captured individuals proved useful for studying the mobility potential of the endangered insect species.     

Effects of Host Interspecific Interaction in the <Emphasis Type="Italic">Maculinea</Emphasis>–<Emphasis Type="Italic">Myrmica</Emphasis> Parasite–Host System

A model of interspecific host competition in a system with one parasite (butterfly—Maculinea) and multiple potential hosts (ants—Myrmica) is presented. Results indicate that host interspecific competition increases the occurrence of multiple host behaviour in Maculinea natural populations but decreases the ability of the parasite populations to adapt to the most abundant host species. These qualitative predictions were compared with data on host specificity, with good agreement. Analysis of the data also indicates that Maculinea teleius and Maculinea arion respond differently to changes in relative host abundances. Maculinea teleius shows a larger fraction of sites where it displays multiple host behaviour and a larger fraction of sites where the niches of the hosts overlap. In some instances, Maculinea teleius is adapted to Myrmica hosts that are present in lower frequencies. Maculinea arion is locally more host-specific and occurs at sites where host interspecific competition is unlikely and is more frequently adapted to the most abundant host species.     

<Emphasis Type="Italic">Maculinea alcon</Emphasis> exploits <Emphasis Type="Italic">Myrmica aloba</Emphasis> in Portugal: unusual host ant species of a myrmecophilous butterfly in a peripheral region

Larvae of the obligate myrmecophilous social parasite Maculinea alcon (Lepidoptera: Lycaenidae) were found exclusively using Myrmica aloba (Hymenoptera: Formicidae) ant hosts in NE-Portugal. This is the first record of the host ant usage of any Maculinea species in Portugal, and of any Maculinea using M. aloba nests. These results on such peripheral European populations confirm that knowledge of the local host ant species is crucial for the successful protection of these endangered butterflies, and vital for examining the evolution of such interactions.