Apollo Butterfly Research Articles

DNA barcodes as a tool in biodiversity research: testing pre-existing taxonomic hypotheses in Delphic Apollo butterflies (Lepidoptera, Papilionidae)

Numerous studies have demonstrated that DNA barcoding is an effective tool for detecting DNA clusters, which can be viewed as operational taxonomic units (OTUs), useful for biodiversity research. Frequently, the OTUs in these studies remained unnamed, not connected with pre-existing taxonomic hypotheses, and thus did not really contribute to feasible estimation of species number and adjustment of species boundaries. For the majority of organisms, taxonomy is very complicated with numerous, often contradictory interpretations of the same characters, which may result in several competing checklists using different specific and subspecific names to describe the same sets of populations. The highly species-rich genus Parnassius (Lepidoptera: Papilionidae) is but one example, such as several mutually exclusive taxonomic systems have been suggested to describe the phenotypic diversity found among its populations. Here we provide an explicit flow chart describing how the DNA barcodes can be combined with the existing knowledge of morphology-based taxonomy and geography (sympatry versus allopatry) of the studied populations in order to support, reject or modify the pre-existing taxonomic hypotheses. We then apply this flow chart to reorganize the taxa within the Parnassius delphius species group, solving long-standing taxonomic problems.

Lesions in the wingless gene of the Apollo butterfly (Parnassius apollo, Lepidoptera: Papilionidae) individuals with deformed or reduced wings, coming from the isolated population in Pieniny (Poland)

Parnassius apollo (Lepidoptera: Papilionidae) is a butterfly species which was common in Europe in 19th century, but now it is considered as near threatened. Various programs devoted to protect and save P. apollo have been established, between others the one in Pieniny National Park (Poland). An isolated population of this butterfly has been restored there from a small group of 20–30 individuals in early 1990s. However, deformations or reductions of wings occur in this population in a relatively large number of insects, and the cause of this phenomenon is not known. In this report, the occurrence of lesions in the wingless (wg) gene is demonstrated in most of tested butterflies with deformed or reduced wings, but not in normal insects. Although the analyses indicated that wg lesion(s) cannot be the sole cause of the deformed or reduced wings in the population of P. apollo from Pieniny, the discovery that this genetic defect occurs in most of malformed individuals, can be considered as an important step in understanding this phenomenon.

Changes is genes coding for laccases 1 and 2 may contribute to deformation and reduction of wings in apollo butterfly (Parnassius apollo, Lepidoptera: Papilionidae) from the isolated population in Pieniny National Park (Poland).

An isolated population of apollo butterfly (Parnassius apollo, Lepidoptera: Papilionidae) occurs in Pieniny National Park (Poland). Deformations and reductions of wings in a relatively large number of individuals from this population is found, yet the reasons for these defects are unknown. During studies devoted to identify cause(s) of this phenomenon, we found that specific regions of genes coding of enzymes laccases 1 and 2 could not be amplified from DNA samples isolated from large fractions of malformed insects while expected PCR products were detected in almost all (with one exception) normal butterflies. Laccases (p-diphenol:dioxygen oxidoreductases) are oxidases containing several copper atoms. They catalyse single-electron oxidations of phenolic or other compounds with concomitant reduction of oxygen to water. In insects, their enzymatic activities were found previously in epidermis, midgut, Malpighian tubules, salivary glands, and reproductive tissues. Therefore, we suggest that defects in genes coding for laccases might contribute to deformation and reduction of wings in apollo butterflies, though it seems obvious that deficiency in these enzymes could not be the sole cause of these developmental improperties in P. apollo from Pieniny National Park.

Persistence in a Two-Dimensional Moving-Habitat Model.

Environmental changes are forcing many species to track suitable conditions or face extinction. In this study, we use a two-dimensional integrodifference equation to analyze whether a population can track a habitat that is moving due to climate change. We model habitat as a simple rectangle. Our model quickly leads to an eigenvalue problem that determines whether the population persists or declines. After surveying techniques to solve the eigenvalue problem, we highlight three findings that impact conservation efforts such as reserve design and species risk assessment. First, while other models focus on habitat length (parallel to the direction of habitat movement), we show that ignoring habitat width (perpendicular to habitat movement) can lead to overestimates of persistence. Dispersal barriers and hostile landscapes that constrain habitat width greatly decrease the population's ability to track its habitat. Second, for some long-distance dispersal kernels, increasing habitat length improves persistence without limit; for other kernels, increasing length is of limited help and has diminishing returns. Third, it is not always best to orient the long side of the habitat in the direction of climate change. Evidence suggests that the kurtosis of the dispersal kernel determines whether it is best to have a long, wide, or square habitat. In particular, populations with platykurtic dispersal benefit more from a wide habitat, while those with leptokurtic dispersal benefit more from a long habitat. We apply our model to the Rocky Mountain Apollo butterfly (Parnassius smintheus).

A lack of Wolbachia-specific DNA in samples from apollo butterfly (Parnassius apollo, Lepidoptera: Papilionidae) individuals with deformed or reduced wings.

Various insects contain maternally inherited endosymbiotic bacteria which can cause reproductive alterations, modulation of some physiological responses (like immunity, heat shock response, and oxidative stress response), and resistance to viral infections. In butterflies, Wolbachia sp. is the most frequent endosymbiont from this group, occurring in about 30 % of species tested to date. In this report, the presence of Wolbachia-specific DNA has been detected in apollo butterfly (Parnassius apollo). In the isolated population of this insect occurring in Pieniny National Park (Poland), malformed individuals with deformed or reduced wings appear with an exceptionally high frequency. Interestingly, while total DNA isolated from most (about 85 %) normal insects contained Wolbachia-specific sequences detected by PCR, such sequences were absent in a large fraction (70 %) of individuals with deformed wings and in all tested individuals with reduced wings. These results indicate for the first time the correlation between malformation of wings and the absence of Wolbachia sp. in insects. Although the lack of the endosymbiotic bacteria cannot be considered as the sole cause of the deformation or reduction of wings, one might suggest that Wolbachia sp. could play a protective role in the ontogenetic development of apollo butterfly.

Successful translocation of the threatened Clouded Apollo butterfly (Parnassius mnemosyne) and metapopulation establishment in southern Finland

Translocations have been advocated as a conservation tool helping species adapt to climate and land-use change, but well-documented examples of invertebrates’ translocations are rare. The paper describes a successful translocation of the threatened Clouded Apollo butterfly (Parnassius mnemosyne) in Finland, compares this to a specific failed translocation, and presents conclusions for conservation planning as to factors contributing to the success. Two apparent key characteristics of the successful translocation were greater abundance of larval resources and less open landscape. The successful site was surrounded by forest, which strongly restricted emigration, crucially supporting the survival of the small initial population. Based on 20 mated females’ translocation in 2000, the local population increased slowly, reaching 600 butterflies in 2011. A large translocation patch together with host-plant abundance enabled successful establishment of the local population. Availability of other suitable grassland patches sufficiently nearby was an additional key characteristic, facilitating the Clouded Apollo’s expansion. However, the expansion rate was low; it took seven years for the butterflies to colonise the five nearest patches, only 10–200m from the translocation patch. By 2013, they had colonised all suitable semi-natural grassland patches within 2km from the translocation site and established a seemingly viable metapopulation with 11 subpopulations. The results point to the significance of local habitat area and landscape quality, along with conditions restricting emigration, in determination of suitable translocation sites.

Ten years of abundance data within a spatial population network of the alpine butterfly, Parnassius smintheus

Spatial‐population networks (metapopulations sensu lato) have distinct properties from isolated populations. Within a population network, the abundance, persistence, and dynamics of local populations are affected by other populations within the network. Similarly, the abundance of local populations within the network has a strong effect on the dynamics and persistence of the entire network. In this data paper, we present ten years (1995–2004) of local‐population abundance and seven years of dispersal data within a spatial‐population network consisting of 21 local populations of the Rocky Mountain Apollo butterfly, Parnassius smintheus. Populations were located within alpine meadows above the treeline (≈2100 m) along Lusk (51.01° N, 114.97° W) and Jumpingpound Ridges (50.95° N, 114.91° W) in the front ranges of the Rocky Mountains in Alberta, Canada. Mark–recapture methods were used to monitor all populations in 1995 and 1996, and most populations in 1997 and from 2001–2004. For these years, abundance was estimated using Craig's method. In other years, population size was estimated via Pollard transects and converted to a common estimate of abundance based on a statistical relationship between transect counts and Craig's estimate. We present multiple estimates of abundance for most populations over the adult flight season each year, and the observed number of emigrants and immigrants over the flight season for populations in years where mark–recapture was conducted. These data should be useful in synthetic studies of factors affecting local‐population abundance within spatial‐population networks, landscape genetics, spatial‐population dynamics, and the persistence of spatial‐population networks.

The complete mitochondrial genome of the endangered Apollo butterfly, Parnassius apollo (Lepidoptera: Papilionidae) and its comparison to other Papilionidae species

The Apollo butterfly, Parnassius apollo is a representative species of the butterfly subfamily Parnassiinae. This charming species is one of the most endangered butterfly species in the world. In this study, we sequenced its complete mitochondrial genome (mitogenome), with the aim of accumulating genetic information for further studies of population genetics and mitogenome evolution in the Papilionidae. The 15,404-bp long mitogenome harbors a typical set of 37 genes and is the largest butterfly mitogenome determined, except for Papilio maraho (16,094bp). Like many other sequenced lepidopteran species, one tRNATrp-like and one tRNALeu(UUR)-like sequences were detected in the AT-rich region. A total of 164bp of non-coding sequences are dispersed in 14 regions throughout the genome. The longest intergenic spacer (68bp) is located between tRNASer(AGN) and tRNAGlu, and is the largest spacer at this location among Papilionidae species. This spacer may have resulted from an 8-fold repetition of a TTTCTTCT motif or a 4-fold repetition of a CTTTATTT motif.

An assessment of direct and indirect effects of climate change for populations of the Rocky Mountain Apollo butterfly (Parnassius smintheus Doubleday)

Abstract Climate change is occurring and insects are responding. Current challenges for ecologists and managers are predicting how organisms will respond to continuing climate change and determining how to mitigate potential negative effects. In contrast to broad scale predictions for climate change involving the distribution of species, in this article we highlight the many ways in which local populations of the Rocky Mountain Apollo butterfly (Parnassius smintheus Doubleday) are predicted to respond to climate change. Using experimental and observational data collected over the past 15 years, we detail both direct and indirect effects. In addition, we identify limitations in our knowledge restricting the ability to predict how populations will respond to climate change. Some changes, such as warmer winter temperatures, may have beneficial effects; however, most of the effects of climate change will be detrimental. Variability in snow cover during the overwintering period and habitat loss due to forest encroachment have the largest potential negative effects.

Treeline proximity alters an alpine plant–herbivore interaction

Rising treeline threatens the size and contiguity of alpine meadows worldwide. As trees encroach into previously open habitat, the movement and population dynamics of above-treeline alpine species may be disrupted. This process is well documented in studies of the Rocky Mountain apollo butterfly (Parnassius smintheus). However, subtler consequences of treeline rise remain poorly understood. In this study, we examine whether treeline proximity affects feeding behaviour of P. smintheus larvae, due to altered habitat affecting the distribution and availability of their host plant, lance-leaved stonecrop (Sedum lanceolatum). Understanding differential larval exploitation of food resources in relation to the treeline is an important step in predicting the consequences of continued treeline rise. Parnassius smintheus larvae feed more intensively on S. lanceolatum away from the treeline despite the relative paucity of hosts in these areas, and despite higher fitness penalties associated with the plant's herbivory-induced chemical defenses. Sedum lanceolatum growing near the treeline is less attractive, and therefore represents a less significant resource for P. smintheus larvae than its abundance might imply. If treeline rise continues, we suggest that this pattern of altered resource exploitation may represent a mechanism by which larvae are adversely affected even while adult movement among and within meadows appears sufficient for maintaining population health, and total host availability seems ample.

Effects of experimental population extinction for the spatial population dynamics of the butterfly Parnassius smintheus

While many studies have examined factors potentially impacting the rate of local population extinction, few experimental studies have examined the consequences of extinction for spatial population dynamics. Here we report results from a large-scale, long-term experiment examining the effects of local population extinction for the dynamics of surrounding populations. From 2001–2008 we removed all adult butterflies from two large, neighboring populations within a system of 17 subpopulations of the Rocky Mountain Apollo butterfly, Parnassius smintheus. Surrounding populations were monitored using individual, mark–recapture methods. We found that population removal decreased immigration to surrounding populations in proportion to their connectivity to the removed populations. Correspondingly, within-generation population abundance declined. Despite these effects, we saw little consistent impact between generations. The extinction rates of surrounding populations were unaffected and local population growth was not consistently reduced by the lack of immigration. The broader results show that immigration affects local abundance within generations, but dynamics are mediated by density-dependence within populations and by broader density-independent factors acting between generations. The loss of immigrants resulting from extinction has little impact on the persistence of local populations in this system.

Olfaction and vision in host plant location by Parnassius apollo larvae: consequences for survival and dynamics

Apollo butterflies in Fennoscandia differ from most butterflies because they oviposit almost randomly, away from the host plant. We studied whether this apparently maladaptive behaviour could be compensated for by high accuracy of newly hatched larvae in locating host plants through either olfactory or visual cues. Olfaction capacity was quantified in a two-choice set-up (host plant versus control) in an olfactometer. Visual capacity was quantified in a small-scale open-air arena with a host plant and a dummy control placed randomly at the edge. We found no evidence that larvae oriented themselves towards the host plant in either of these experimental set-ups; instead, larvae moved in the direction of the sun, and arrested movement in the presence of a smell common to the control and treatment of the olfaction test. Because larvae do not locate their host plant at a distance, the probability of finding a host plant is likely to be critically dependent on host plant density. We tested this assumption by releasing larvae in open-air arenas with three different host plant densities. The probability of larvae reaching a host plant depended strongly on host plant density. A comparison with host plant densities observed in nature suggests that Apollo butterfly larvae require a relatively high host plant density in order to have a reasonable probability of locating a host plant before perishing. Our findings provide a clear proximate link between host plant density and larval mortality, a presumed key factor affecting population processes in the Apollo butterfly.

Local extinction synchronizes population dynamics in spatial networks

Spatial population theory predicts that synchrony in the dynamics of local populations should decrease as dispersal among populations decreases. Thus, it would be expected that the extinction of local populations and the attendant loss of immigrants to surrounding populations would reduce synchrony. We tested this hypothesis through a large-scale experiment, simulation of the experimental system and general models. Experimental removal of two adjacent subpopulations of the Rocky Mountain Apollo butterfly, Parnassius smintheus within a network consisting of 15 other local populations resulted in a decrease in immigration to surrounding populations that was proportional to their connectivity to the removal populations. These populations also showed a significant increase in synchrony during population removal. The spatial extent of the synchrony showed good agreement with the predicted loss of immigrants owing to the removals. Simulation of the Parnassius system showed a similar short-term result and also indicated that permanent loss of populations produces structural changes increasing synchrony. General models indicate that an increase in synchrony following extinction occurs when populations undergoing extinction have different carrying capacities than surrounding populations. The result is not owing to biased migration per se, but rather is because of the number of immigrants relative to the carrying capacity. Synchrony following extinction should be most common for patchy populations, but can occur in any situation where carrying capacities differ. Overall, our results indicate that local extinction can create a positive feedback for extinction risk, increasing the probability of extinction for population networks by synchronizing their dynamics.

Resources influence dispersal and population structure in an endangered butterfly

Abstract. 1. The degree of dispersal between patches in a population partly determines its structure. Mark–release–recapture (MRR) studies of an archipelago population of the endangered apollo butterfly (Parnassius apollo, L 1758) revealed that most (79%) adult individuals remained within the patch where they were marked.2. Emigration out of a patch was female‐biased and decreased with the amount of (adult) nectar resources on a patch, but increased with greater proximity to nectar on other patches. Local population numbers positively correlated with total numbers of both emigrants and immigrants. Host‐plant abundance (larval resource) did not influence dispersal.3. The dispersal and population structure of this archipelago population differs pronouncedly from that of a previously studied mainland population where between‐patch dispersal is more frequent even when correcting for differences in habitat structure between the populations, and consequently the population structure is more patchy or open.4. The most prominent environmental factor affecting emigration is nectar resources. Both local patch conditions in terms of nectar on the patch and nectar availability on surrounding patches are important.5. The most important implications for conservation are that resource distribution affects dispersal and hence population‐level processes such as structure, and that emigration and immigration should be viewed as separate processes potentially governed by different environmental factors.

Effectiveness of population recovery projects based on captive breeding

A recovery programme for a population of Apollo butterflies in the Pieniny National Park was monitored over 12 years in order to assess its effectiveness. The programme was based on population ecology and metapopulation theory, and its object was to replenish wild populations with captive-reared individuals. Thanks to the adopted measures, the local population initially numbering only 30–50 individuals increased to ca. 1000 imagines, forming a functional metapopulation. Nevertheless, analysis of population dynamics parameters indicated that only some of the subpopulations were stable; a significant percentage of the subpopulation fractions represented sink populations whose survival depended either on an external supply of captive-reared individuals or on individuals from other source subpopulations. In some cases, intensively supplying the subpopulation with captive-reared individuals resulted in a decrease of that deme’s abundance. The results suggest that supplying captive-reared individuals to an endangered population may entail some risks and should be done with caution.

Accounting for possible detectable distances in a comparison of dispersal: Apollo dispersal in different habitats

Estimates of dispersal distances, when studied using mark-release-recapture techniques, are affected by many extrinsic factors such as study area size, local population density, and landscape configuration. We show that a comparison of observed dispersal distances between groups (e.g. sexes, populations, study periods) may lead to erroneous conclusions when groups differ in their distribution of possible detectable distances (the distribution of all distances in the system weighted by the number of marked individuals that have the possibility to move those distances). We show how this distribution can be used to: (1) test whether the scale of the study area is adequate for a description of mean dispersal; (2) express dispersal as a fraction of the number observed over the number of possible detectable movements for each distance. Dispersal scaled in this fashion can be compared between groups using standard statistical techniques. For illustration, we analyse five mark-release-recapture studies on the Apollo butterfly Parnassius apollo from two populations. Correcting for possible detectable distances shows that dispersal in an archipelago population is lower than in a coastal population, possibly due to harsher habitat between the patches in the former population.

Midgut protease activities in monophagous larvae of Apollo butterfly, Parnassius apollo ssp. frankenbergeri

We assayed the relative activities of midgut proteolytic enzymes in individuals of the fourth (L 4) and fifth (L 5) instar of Apollo larvae, inhabiting Pieniny Mts (southern Poland). The comparisons between midgut tissue with glicocalyx (MT) and liquid midgut contents with peritrophic membrane (MC) were made. Optimal media pHs of the assayed proteolytic enzymes in P. apollo midgut samples were similar to those of other lepidopteran species. Endopeptidases, as well as carboxypeptidases, digested effectively in alkaline environment, while aminopeptidases were active in a broad pH range. Trypsin is probably the main endoprotease (correlation with caseinolytic activity in MC of L 5 larvae: r = 0.606 ; p = 0.004 ); however, its activity was low as compared with that in other leaf-eating Lepidoptera. This suggests a minor role of trypsin and chymotrypsin in protein digestion in Apollo larvae, probably due to limited availability of the leaf proteins. Instead, due to very high carboxypeptidase A activity in midgut tissue, the larvae obtain exogenous amino acids either directly or from oligopeptides and glycoproteins. High and significant positive correlations between the enzyme activity and glucosidase as well as galactosidase activities strongly support this opinion. To cite this article: M. Nakonieczny et al., C. R. Biologies 330 (2007).

Midgut glycosidases activities in monophagous larvae of Apollo butterfly, Parnassius apollo ssp. frankenbergeri

Parnassius apollo (Lepidoptera, Papilionidae) declines on numerous localities all over Europe. Its local subspecies frankenbergeri, inhabiting the Pieniny Mts (southern Poland) and successfully recovered from extinction, is monophagous in larval stage. In natural conditions, it completes development on the orpine Sedum telephium ssp. maximum. Since proper quality and quantity of necessary nutritional compounds of the food plant ensure developmental success, the digestive processes in the insect midgut should reflect adaptation to a specific food source. The paper presents, for the first time, the activity of detected glycolytic enzymes in midgut tissue and liquid gut contents of the L 4 and L 5 instars of P. apollo larvae. α-Amylase plays the main role in utilization of carbohydrates, contrary to cellulase activity. Saccharase seems to be the main disaccharidase, and high activity of β-glycosidase enables hydrolysis of the plant glycosides. Trehalase activity was unexpectedly low and comparable to those of cellobiase and lactase. α-Amylolytic and other glycolytic activities indicate that larvae utilize starch and other carbohydrate compounds as energy sources. Possible use of some plant allelochemicals as energy sources by Apollo larvae is discussed. To cite this article: M. Nakonieczny et al., C. R. Biologies 329 (2006).

Modelling the spatial distribution of a threatened butterfly: Impacts of scale and statistical technique

This paper compares the performance of five modelling methods in the prediction of a species distribution, using a data set describing the distribution of the threatened clouded apollo butterfly ( Parnassius mnemosyne) in south-west Finland. The five statistical techniques included were: generalized linear models (GLM), generalized additive models (GAM), classification tree analysis (CTA), neural networks (ANN) and multiple adaptive regression splines (MARS). The accuracy of the models was examined at three spatial resolutions (1, 25 and 100 ha) by area under the curve (AUC) and kappa statistics. All five modelling techniques had a relatively high discrimination capacity for the occurrence of clouded apollo. Classification tree analysis provided the least robust model performance. The differences between the other methods were small, although GAM and MARS provided marginally the best stability and performance. The most accurate models were developed for the resolutions of 1 ha (highest AUC values) and 25 ha (highest kappa values) and the least accurate models for the resolution of 100 ha. Our work shows that modern modelling techniques can provide useful forecasts of species distributions in unsurveyed parts of landscapes and provide valuable contributions to conservation and management planning. However, the success of applying the new modelling tools can be influenced by the choice of statistical technique and especially of spatial resolution. In conclusion, small changes in the spatial scale may result in a clear decrease in the model performance and thus caution should be exercised when implementing the models and their predictions in practice.

Consequences of the spatial configuration of resources for the distribution and dynamics of the endangered Parnassius apollo butterfly

Based on several years of data from two populations of the endangered Apollo butterfly ( Parnassius apollo, L. 1758), we study how the amount and spatial location of patches of larval (host-plant) and adult (nectar plant) resources affects the distribution of females and their larval offspring in the following year. In the coastal population (nectar-plant and host-plant patches spatially segregated), females moved frequently between patches to aggregate on larger host-plant patches close to nectar-plant patches. In the archipelago population (where nectar-plants and host-plants co-occur), the abundance of females increased with higher proximity to other host-plant patches and with more nectar-plants on the patch. Next year’s larval abundance correlated with the abundance of females in the previous season in both populations. A model of the population dynamics in the two populations in relation to the spatial configuration of nectar and host-plant patches showed that the spatial configuration of larval and adult resources had population-dynamical consequences. In many organisms, different life-history stages use different resources. Incorporation of information on the location and abundance of adult resources can provide additional insight for the suitability of a particular landscape in harbouring a population.