Combining environmental niche models, multi-grain analyses, and species traits identifies pervasive effects of land use on butterfly biodiversity across Italy.

Understanding how species respond to human activities is paramount to ecology and conservation science, one outstanding question being how large-scale patterns in land use affect biodiversity. To facilitate answering this question, we propose a novel analytical framework that combines environmental niche models, multi-grain analyses, and species traits. We illustrate the framework capitalizing on the most extensive dataset compiled to date for the butterflies of Italy (106,514 observations for 288 species), assessing how agriculture and urbanization have affected biodiversity of these taxa from landscape to regional scales (3-48 km grains) across the country while accounting for its steep climatic gradients. Multiple lines of evidence suggest pervasive and scale-dependent effects of land use on butterflies in Italy. While land use explained patterns in species richness primarily at grains ≤12 km, idiosyncratic responses in species highlighted "winners" and "losers" across human-dominated regions. Detrimental effects of agriculture and urbanization emerged from landscape (3-km grain) to regional (48-km grain) scales, disproportionally affecting small butterflies and butterflies with a short flight curve. Human activities have therefore reorganized the biogeography of Italian butterflies, filtering out species with poor dispersal capacity and narrow niche breadth not only from local assemblages, but also from regional species pools. These results suggest that global conservation efforts neglecting large-scale patterns in land use risk falling short of their goals, even for taxa typically assumed to persist in small natural areas (e.g., invertebrates). Our study also confirms that consideration of spatial scales will be crucial to implementing effective conservation actions in the Post-2020 Global Biodiversity Framework. In this context, applications of the proposed analytical framework have broad potential to identify which mechanisms underlie biodiversity change at different spatial scales.

Genotoxic effects of particulate matter on larvae of a common and widespread butterfly along an urbanization gradient.

Biodiversity is currently declining worldwide. Several threats have been identified such as habitat loss and climate change. It is unknown if and how air pollution can work in addition or in synergy to these threats, contributing to the decline of current species and/or local extinction. Few studies have investigated the effects of particulate matter (PM), the main component of air pollution, on insects, and no studies have investigated its genotoxic effects through Micronucleus assay. Butterflies play an important role in the environment, as herbivores during larval stages, and as pollinators as adults. The aim of this study was to evaluate the genotoxic effects of PM10 from different sites along a gradient of population urbanization, on a common cabbage butterfly species (Pieris brassicae). PM10 was collected from April to September in an urban (Turin, Italy), a suburban (Druento, Italy) and a mountain site (Ceresole Reale, Italy) with different urbanization levels. P. brassicae larvae (n = 218) were reared in the laboratory under controlled conditions (26 °C, L:D 15:9) on cabbage plants (average 9.2 days), and they were exposed to PM10 organic extracts (20 and 40 m3/mL) or dimethyl sulfoxide (controls) through vaporization. After exposure, larvae were dissected and cells were used for the Micronucleus (MN) assay. Results showed that all PM extracts induced significant DNA damage in exposed larvae compared to controls, and that increasing the PM dose (from 20 to 40 m3/mL) increased genotoxic effects. However, we did not detect any significant differences between sites with different urbanization levels. In conclusion, PM at different concentrations induced genotoxic effects on larvae of a common butterfly species. More alarmingly, PM could work in addition to and/or in synergy with other compounds (e.g. pesticides) and, especially on species already threatened by other factors (e.g. fragmentation), thus affecting the vitality of populations, leading to local extinctions.

Two ways to be endemic. Alps and Apennines are different functional refugia during climatic cycles.

Endemics co-occur because they evolved in situ and persist regionally or because they evolved ex situ and later dispersed to shared habitats, generating evolutionary or ecological endemicity centres, respectively. We investigate whether different endemicity centres can intertwine in the region ranging from Alps to Sicily, by studying their butterfly fauna. We gathered an extensive occurrence data set for butterflies of the study area (27,123 records, 269 species, in cells of 0.5 × 0.5 degrees of latitude-longitude). We applied molecular-based delimitation methods (GMYC model) to 26,557 cytochrome c oxidase subunit 1 (COI) sequences of Western Palearctic butterflies. We identified entities based on molecular delimitations and/or the checklist of European butterflies and objectively attributed occurrences to their most probable entity. We obtained a zoogeographic regionalisation based on the 69 endemics of the area. Using phylogenetic ANOVA we tested if endemics from different centres differ from each other and from nonendemics for key ecological traits and divergence time. Endemicity showed high incidence in the Alps and Southern Italy. The regionalisation separated the Alps from the Italian Peninsula and Sicily. The endemics of different centres showed a high turnover and differed in phylogenetic distances, phenology and distribution traits. Endemics are on average younger than nonendemics and the Peninsula-Sicily endemics also have lower variance in divergence than those from the Alps. The observed variation identifies Alpine endemics as paleoendemics, now occupying an ecological centre, and the Peninsula-Sicily ones as neoendemics, that diverged in the region since the Pleistocene. The results challenge the common view of the Alpine-Apennine area as a single "Italian refugium".

How long is 3 km for a butterfly? Ecological constraints and functional traits explain high mitochondrial genetic diversity between Sicily and the Italian Peninsula.

Populations inhabiting Mediterranean islands often show contrasting genetic lineages, even on islands that were connected to the mainland during glacial maxima. This pattern is generated by forces acting in historical and contemporary times. Understanding these phenomena requires comparative studies integrating genetic structure, functional traits and dispersal constraints. Using as a model the butterfly species living across the Messina strait (3 km wide) separating Sicily from the Italian Peninsula, we aimed to unravel the mechanisms limiting the dispersal of matrilines and generating genetic differentiation across a narrow sea strait. We analysed the mitochondrial COI gene of 84 butterfly species out of 90 documented in Sicily and compared them with populations from the neighbouring southern Italian Peninsula (1,398 sequences) and from the entire Palaearctic region (8,093 sequences). For each species, we regressed 13 functional traits and 2 ecological constraints to dispersal (winds experienced at the strait and climatic suitability) against genetic differentiation between Sicily and Italian Peninsula to understand the factors limiting dispersal. More than a third of the species showed different haplogroups across the strait and most of them also represented endemic haplogroups for this island. One fifth of Sicilian populations (and 32.3% of endemic lineages) had their closest relatives in distant areas, instead of the neighbouring Italian Peninsula, which suggests high relictuality. Haplotype diversity was significantly explained by the length of the flight period, an intrinsic phenology trait, while genetic differentiation was explained by both intrinsic traits (wingspan and degree of generalism) and contemporary local constraints (winds experienced at the strait and climatic suitability). A relatively narrow sea strait can produce considerable differentiation among butterfly matrilines and this phenomenon showed a largely deterministic fingerprint. Because of unfavourable winds, populations of the less dispersive Sicilian butterflies tended to differentiate into endemic variants or to maintain relict populations. Understanding these phenomena required the integration of DNA sequences, species traits and physical constraints for a large taxon at continental scale. Future studies may reveal if the patterns here shown for mitochondrial DNA are also reflected in the nuclear genome or, alternatively, are the product of limited female dispersal.

A micronucleus assay detects genotoxic effects of herbicide exposure in a protected butterfly species.

Lycaena dispar Hawort (Lepidoptera: Lycaenidae), a protected butterfly, is declining in Europe, but it thrives in rice fields in northern Italy. Here, agrochemical usage could threaten its long-term survival. We investigated, by micronucleus (MN) assay, the genotoxic effect of glyphosate, a common herbicide, on L. dispar larvae. Micronuclei (MNi) are DNA fragments separated from the main nucleus and represent the result of genomic damage that has been transmitted to daughter cells. In a control/treatment experiment, we extracted epithelial cells from last-instar larvae fed with Rumex spp. plants sprayed with a solution containing 3.6 g/L of glyphosate, and from larvae fed with unsprayed plants. MNi and other chromosomal aberrations-nuclear buds (NBUDs) and bi-nucleated cells-were then scored in 1000 cells/subject. Significant differences were found between glyphosate-exposed and control groups in terms of MNi and total genomic damage, but not in terms of NBUDs or bi-nucleated cells. We reported a possible genomic damage induced by glyphosate on larvae of L. dispar. For the first time, a MN assay was used in order to evaluate the genomic damage on a phytophagous invertebrate at the larval stage. Increased levels of MNi reflect a condition of genomic instability that can result in reduced vitality and in an increased risk of local extinction. Therefore, farmland management compatible with wildlife conservation is needed.

Effects of Host Interspecific Interaction in the Maculinea-Myrmica 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.

Plant defences against ants provide a pathway to social parasitism in butterflies.

Understanding the chemical cues and gene expressions that mediate herbivore-host-plant and parasite-host interactions can elucidate the ecological costs and benefits accruing to different partners in tight-knit community modules, and may reveal unexpected complexities. We investigated the exploitation of sequential hosts by the phytophagous-predaceous butterfly Maculinea arion, whose larvae initially feed on Origanum vulgare flowerheads before switching to parasitize Myrmica ant colonies for their main period of growth. Gravid female butterflies were attracted to Origanum plants that emitted high levels of the monoterpenoid volatile carvacrol, a condition that occurred when ants disturbed their roots: we also found that Origanum expressed four genes involved in monoterpene formation when ants were present, accompanied by a significant induction of jasmonates. When exposed to carvacrol, Myrmica workers upregulated five genes whose products bind and detoxify this biocide, and their colonies were more tolerant of it than other common ant genera, consistent with an observed ability to occupy the competitor-free spaces surrounding Origanum. A cost is potential colony destruction by Ma. arion, which in turn may benefit infested Origanum plants by relieving their roots of further damage. Our results suggest a new pathway, whereby social parasites can detect successive resources by employing plant volatiles to simultaneously select their initial plant food and a suitable sequential host.

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.

The "Second Life" of laboratory rats (Rattus norvegicus): Assessment of social behavior of a colony of rats based on social network analysis.

Rattus norvegicus is a social animal and holds a significant economic value, considering its use in scientific research. Here, we use the Social Network Analysis (SNA) approach to study the social interactions of a group of rats held in a post-laboratory animal care facility. We collected interaction data during four study periods, for a total of 60 days. At the group level, rats presented two communities for each study period, consisting mainly of littermates. At individual level, we found that the rats preferred to interact with individuals of the same strain and laboratory of origin and with littermates. At temporal level, we studied how stable social interactions were over time. During the first study period, we found high social stability, whereas the introduction of new individuals in the subsequent period caused social rearrangements; however, the initial social stability was restored. Our findings have shown that studying the social behavior of rats using SNA is a valuable tool for advancing our understanding of the social system of this species, which has the potential to enhance management and welfare practices.

Cabbage butterfly as bioindicator species to investigate the genotoxic effects of PM10.

Atmospheric pollution poses a serious threat to environment and human health, and particulate matter (PM) is one of the major contributors. Biological effects induced by PM are investigated through in vitro assays using cells and by in vivo tests with laboratory model animals. However, also the estimation of adverse effects of pollutants, including airborne ones, on wild animals, such as insects, is an essential component of environmental risk assessment. Among insects, butterflies are sensitive to environmental changes and are important wild pollinators, so they might be suitable as environmental bioindicator species. The aim of this study was to evaluate the suitability of a wild cabbage butterfly species (Pieris brassicae) as a bioindicator organism to assess the genotoxic effects of PM10 collected in different sites. PM10 was collected from April to September in urban, suburban, and rural sites. P. brassicae larvae were reared in laboratory under controlled conditions on cabbage plants and exposed to PM10 organic extracts or dimethyl sulfoxide (controls) through vaporization. After exposure, larvae were dissected, and cells were used for comet assay. All PM extracts induced significant DNA damage in exposed larvae compared to controls and the extract collected in the most polluted site caused the highest genotoxic effect. In conclusion, the study suggested that butterflies, such as P. brassicae, could be applied as sensitive and promising bioindicators to investigate air quality and PM genotoxicity. Indeed, the use of these organisms allows the detection of genotoxic effects induced by PM sampled also in low-polluted areas.

Distribution Drivers of the Alien Butterfly Geranium Bronze (Cacyreus marshalli) in an Alpine Protected Area and Indications for an Effective Management.

Cacyreus marshalli is the only alien butterfly in Europe. It has recently spread in the Gran Paradiso National Park (GPNP), where it could potentially compete with native geranium-consuming butterflies. Our study aimed to (1) assess the main drivers of its distribution, (2) evaluate the potential species distribution in GPNP and (3) predict different scenarios to understand the impact of climate warming and the effect of possible mitigations. Considering different sampling designs (opportunistic and standardised) and different statistical approaches (MaxEnt and N-mixture models), we built up models predicting habitat suitability and egg abundance for the alien species, testing covariates as bioclimatic variables, food plant (Pelargonium spp.) distribution and land cover. A standardised approach resulted in more informative data collection due to the survey design adopted. Opportunistic data could be potentially informative but a major investment in citizen science projects would be needed. Both approaches showed that C. marshalli is associated with its host plant distribution and therefore confined in urban areas. Its expansion is controlled by cold temperatures which, even if the host plant is abundant, constrain the number of eggs. Rising temperatures could lead to an increase in the number of eggs laid, but the halving of Pelargonium spp. populations would mostly mitigate the trend, with a slight countertrend at high elevations.

Zerynthia polyxena Locally Monophagous on Aristolochia pallida in the Susa Valley.

In insect-plant biology, oviposition choices and larval development on different host plants are crucial factors to be investigated. To design conservation strategies for protected Lepidoptera, which are overall oligophagous but locally monophagous, it is important to understand which host plant species is locally preferred. We thus investigated oviposition choices and larval development of the protected butterfly Zerynthia polyxena in controlled laboratory conditions, using three possible host plant species which are present in the Piedmont region: Aristolochia pallida, A. clematitis, and A. rotunda. We found that laboratory conditions are not favorable for Z. polyxena oviposition, even if the fertility of the females was in normal range for Papilionidae. However, we were able to understand the local monophagy of the species on A. pallida in the Susa Valley through larval survival and development stages. Egg hatching was similar among the three host plant species; however, even if larvae eat and grow similarly on the different host plant species until the third larval stage, the only larvae that reached the pupal stage were those fed with A. pallida. In conclusion, whereas Z. polyxena is oligophagous in the rest of Europe for the genus Aristolochia, the species is locally monophagous on A. pallida in the Susa Valley.

Changes in Alpine Butterfly Communities during the Last 40 Years.

Our work aims to assess how butterfly communities in the Italian Maritime Alps changed over the past 40 years, in parallel with altitudinal shifts occurring in plant communities. In 2019, we sampled butterflies at 7 grassland sites, between 1300-1900 m, previously investigated in 2009 and 1978, by semi-quantitative linear transects. Fine-scale temperature and precipitation data elaborated by optimal interpolation techniques were used to quantify climate changes. The changes in the vegetation cover and main habitat alterations were assessed by inspection of aerial photographs (1978-2018/1978-2006-2015). The vegetation structure showed a marked decrease of grassland habitats and an increase of woods (1978-2009). Plant physiognomy has remained stable in recent years (2009-2019) with some local exceptions due to geomorphic disturbance. We observed butterfly 'species substitution' indicating a general loss in the more specialised and a general gain in more tolerant elements. We did not observe any decrease in species richness, but rather a change in guild compositions, with (i) an overall increased abundance in some widespread and common lowland species and (ii) the disappearance (or strong decrease) of some alpine (high elevation) species, so that 'resilience' could be just delusive. Changes in butterfly community composition were consistent with predicted impacts of local warming.

Stabilisation of some names of European butterflies (Lepidoptera: Pieridae) in their prevailing usage.

To stabilise nomenclature in its prevailing usage, the authors designate three lectotypes, respectively for the nominal taxa Pontia daplidice race nitida Verity, 1908 (Museo di Storia naturale della Università di Firenze, sezione di Zoologia, 'La Specola' MZUF), for Euchloe crameri Butler, 1869 (Natural History Museum UK, NHMUK) and for Euchloe ausonia var. esperi Kirby, 1871 (National Museum of Ireland, NMI). They also designate two neotypes, respectively for [Papilio] ausonia Hübner, [1804] (MZUF) and for Pontia simplonia Freyer, 1829 (Jura Museum, Eichstätt, JME).

A new comprehensive trait database of European and Maghreb butterflies, Papilionoidea.

Trait-based analyses explaining the different responses of species and communities to environmental changes are increasing in frequency. European butterflies are an indicator group that responds rapidly to environmental changes with extensive citizen science contributions to documenting changes of abundance and distribution. Species traits have been used to explain long- and short-term responses to climate, land-use and vegetation changes. Studies are often characterised by limited trait sets being used, with risks that the relative roles of different traits are not fully explored. Butterfly trait information is dispersed amongst various sources and descriptions sometimes differ between sources. We have therefore drawn together multiple information sets to provide a comprehensive trait database covering 542 taxa and 25 traits described by 217 variables and sub-states of the butterflies of Europe and Maghreb (northwest Africa) which should serve for improved trait-based ecological, conservation-related, phylogeographic and evolutionary studies of this group of insects. We provide this data in two forms; the basic data and as processed continuous and multinomial data, to enhance its potential usage.

Relative importance of density-dependent regulation and environmental stochasticity for butterfly population dynamics.

The relative contribution of density-dependent regulation and environmental stochasticity to the temporal dynamics of animal populations is one of the central issues of ecology. In insects, the primary role of the latter factor, typically represented by weather patterns, is widely accepted. We have evaluated the impact of density dependence as well as density-independent factors, including weather and mowing regime, on annual fluctuations of butterfly populations. As model species, we used Maculinea alcon and M. teleius living in sympatry and, consequently, we also analysed the effect of their potential competition. Density dependence alone explained 62 and 42% of the variation in the year-to-year trends of M. alcon and M. teleius, respectively. The cumulative Akaike weight of models with density dependence, which can be interpreted as the probability that this factor should be contained in the most appropriate population dynamics model, exceeded 0.97 for both species. In contrast, the impacts of inter-specific competition, mowing regime and weather were much weaker, with their cumulative weights being in the range of 0.08-0.21; in addition, each of these factors explained only 2-5% of additional variation in Maculinea population trends. Our results provide strong evidence for density-dependent regulation in Maculinea, while the influence of environmental stochasticity is rather minor. In the light of several recent studies on other butterflies that detected significant density-dependent effects, it would appear that density-dependent regulation may be more widespread in this group than previously thought, while the role of environmental stochasticity has probably been overestimated. We suggest that this misconception is the result of deficiencies in the design of most butterfly population studies in the past, including (1) a strong focus on adults and a neglect of the larval stage in which density-dependent effects are most likely to occur; (2) an almost exclusive reliance on transect count results that may confound the impact of environmental stochasticity on butterfly numbers with its impact on adult longevity.

Chemical, Physiological and Molecular Responses of Host Plants to Lepidopteran Egg-Laying.

Plant-lepidopteran interactions involve complex processes encompassing molecules and regulators to counteract defense responses they develop against each other. Lepidoptera identify plants for oviposition and exploit them as larval food sources to complete their development. In turn, plants adopt different strategies to overcome and limit herbivorous damages. The insect egg deposition on leaves can already induce a number of defense responses in several plant species. This minireview deals with the main features involved in the interaction between plants and lepidopteran egg-laying, focusing on responses from both insect and plant side. We discuss different aspects of direct and indirect plant responses triggered by lepidopteran oviposition. In particular, we focus our attention on the mechanisms underlying egg-induced plant defenses that can i) directly damage the eggs such as localized hypersensitive response (HR)-like necrosis, neoplasm formation, production of ovicidal compounds and ii) indirect defenses, such as production of oviposition-induced plant volatiles (OIPVs) used to attract natural enemies (parasitoids) able to kill the eggs or hatching larvae. We provide an overview of chemical, physiological, and molecular egg-mediated plant responses induced by both specialist and generalist lepidopteran species, also dealing with effectors, elicitors, and chemical signals involved in the process. Egg-associated microorganisms are also discussed, although little is known about this third partner participating in plant-lepidopteran interactions.

Host specificity pattern and chemical deception in a social parasite of ants.

In natural ecosystems, relationships between organisms are often characterised by high levels of complexity, where vulnerabilities in multi-trophic systems are difficult to identify, yet variation in specific community modules can be traceable. Within the complex community interactions, we can shed new light on dynamics by which co-evolutionary outcomes can inform science-led conservation. Here we assessed host-ant use in six populations of the butterfly Phengaris (=Maculinea) rebeli, an obligate social parasite of Myrmica ants and a model system in evolutionary and conservation ecology. Starting from the initial distribution of eggs, we estimated the survival of the parasite in the wild in nests of seven Myrmica ant species, and analysed the chemical cues evolved by the parasites to subvert its host defences. We found local variations in host specificity that are consistent with similarities found in the chemical profiles of hosts and parasites on different sites. At some sites, only one ant species is successfully exploited; at others, multiple-host populations are used. Understanding how stable or adaptable these associations are is essential knowledge when devising conservation measures to maintain keystone species of ant and locally adapted populations of Phengaris butterfly species, which are rare, threatened and a high priority for conservation worldwide.

Integrating three comprehensive data sets shows that mitochondrial DNA variation is linked to species traits and paleogeographic events in European butterflies.

Understanding the dynamics of biodiversity, including the spatial distribution of genetic diversity, is critical for predicting responses to environmental changes, as well as for effective conservation measures. This task requires tracking changes in biodiversity at large spatial scales and correlating with species functional traits. We provide three comprehensive resources to understand the determinants for mitochondrial DNA differentiation represented by (a) 15,609 COI sequences and (b) 14 traits belonging to 307 butterfly species occurring in Western-Central Europe and (c) the first multi-locus phylogenetic tree of all European butterfly species. By applying phylogenetic regressions we show that mitochondrial DNA spatial differentiation (as measured with GST , G'ST , D and DST ) is negatively correlated with species traits determining dispersal capability and colonization ability. Thanks to the high spatial resolution of the COI data, we also provide the first zoogeographic regionalization maps based on intraspecific genetic variation. The overall pattern obtained by averaging the spatial differentiation of all Western-Central European butterflies shows that the paradigm of long-term glacial isolation followed by rapid pulses of post-glacial expansion has been a pervasive phenomenon in European butterflies. The results and the extensive data sets we provide here constitute the basis for genetically-informed conservation plans for a charismatic group in a continent where flying insects are under alarming decline.

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'.