Geometric Morphometric Framework Research Articles

Hide and seek shark teeth in Random Forests: machine learning applied to Scyliorhinus canicula populations.

Shark populations that are distributed alongside a latitudinal gradient often display body size differences at sexual maturity and vicariance patterns related to their number of tooth files. Previous works have demonstrated that Scyliorhinus canicula populations differ between the northeastern Atlantic Ocean and the Mediterranean Sea based on biological features and genetic analysis. In this study, we sample more than 3,000 teeth from 56 S. canicula specimens caught incidentally off Roscoff and Banyuls-sur-Mer. We investigate population differences based on tooth shape and form by using two approaches. Classification results show that the classical geometric morphometric framework is outperformed by an original Random Forests-based framework. Visually, both S. canicula populations share similar ontogenetic trends and timing of gynandric heterodonty emergence but the Atlantic population has bigger, blunter teeth, and less numerous accessory cusps than the Mediterranean population. According to the models, the populations are best differentiated based on their lateral tooth edges, which bear accessory cusps, and the tooth centroid sizes significantly improve classification performances. The differences observed are discussed in light of dietary and behavioural habits of the populations considered. The method proposed in this study could be further adapted to complement DNA analyses to identify shark species or populations based on tooth morphologies. This process would be of particular interest for fisheries management and identification of shark fossils.

Variation in shell morphology of the European pond turtle,Emys orbicularis, in fragmented central European populations

AbstractThe European pond turtle (Emys orbicularis) is a widely distributed freshwater species inhabiting much of Europe, but it is often in population decrease or is locally extinct. In this study, we sampled five central European populations, of which four were autochthonous and one was introduced outside the native range. Moreover, two of the native populations were relatively isolated and at the periphery of the species, range. Using the frequency of shell anomalies, a geometric morphometric framework and an analysis of fluctuating asymmetry, we aimed to determine the degree of morphological differentiation among different populations. Significantly, a smaller number of individuals with a malformed shell or scutes occurred in the native core range population, which potentially has a high level of gene flow (Hungary). Although neither canonical variate analysis nor the morphological disparity analysis showed distinct differences between populations, we found significantly higher shell asymmetry in the two marginal populations (Austria and Slovakia) compared with the core range populations. Our results might thus support the central–marginal hypothesis and indicate potential genetically based conservation problems owing to demographic bottlenecks and/or isolation in marginal populations.

Integrative taxonomy and species delimitation of Rhipicephalus turanicus (Acari: Ixodida: Ixodidae)

Rhipicephalus turanicus ticks are widely distributed across the Palearctic and Afrotropics. These two continental populations display differences in morphological characters that raise the question of a potential species boundary. However, the taxonomic status of these morphologically divergent lineages is uncertain because R. turanicus from Cyprus and Zambia have been shown to interbreed and produce fertile hybrids. We employ integrative taxonomy that considers data from mtDNA sequences (12S and 16S rDNA), geographic distribution, traditional (qualitative) morphology, as well as shape outlines of female spiracles and male adanal plates measured in a geometric morphometric framework (quantitative morphology) to resolve this taxonomic issue. Molecular lines of evidence (12S and 16S rDNA) support taxonomic separation between ticks sampled in the Afrotropics and the Palearctic. This is corroborated by qualitative and quantitative morphology. Within the Palearctic, two sub-lineages were recovered based on sequence data that loosely correspond to southern Europe and the Middle East/Asia. One new species, Rhipicephalus afranicus n. sp. is described from South Africa with a geographic distribution that extends into eastern Africa. This leaves R. turanicus sensu lato comprised of two lineages located in southern Europe and the Middle East/Asia. The type locality for R. turanicus is in Uzbekistan, thus the Middle East/Asia lineage is considered R. turanicus sensu stricto. Detailed descriptions are provided for R. afranicus n. sp. and R. turanicus sensu stricto together with high resolution images. Speciation is attributed to recent Sahara desert expansion that formed a natural barrier to dispersal approximately 5–7 million years ago. However, reproductive potential between these two species suggests that divergence time and mode of speciation were not sufficient for the development of reproductive isolation. We suggest speciation was complicated by divergence and population reintegration events driven by oscillating climatic conditions contributing to reticulate evolution and maintenance of compatibility between reproductive mechanisms. This study represents an integrative (iterative) approach to delimiting Rhipicephalus spp., and provides the first application of shape outlines for female spiracles and male adanal plates measured in a geometric morphometric framework, applied to testing species boundaries between ticks.

Are endocasts good proxies for brain size and shape in archosaurs throughout ontogeny?

Cranial endocasts, or the internal molds of the braincase, are a crucial correlate for investigating the neuroanatomy of extinct vertebrates and tracking brain evolution through deep time. Nevertheless, the validity of such studies pivots on the reliability of endocasts as a proxy for brain morphology. Here, we employ micro‐computed tomography imaging, including diffusible iodine‐based contrast‐enhanced CT, and a three‐dimensional geometric morphometric framework to examine both size and shape differences between brains and endocasts of two exemplar archosaur taxa – the American alligator (Alligator mississippiensis) and the domestic chicken (Gallus gallus). With ontogenetic sampling, we quantitatively evaluate how endocasts differ from brains and whether this deviation changes during development. We find strong size and shape correlations between brains and endocasts, divergent ontogenetic trends in the brain‐to‐endocast correspondence between alligators and chickens, and a comparable magnitude between brain–endocast shape differences and intraspecific neuroanatomical variation. The results have important implications for paleoneurological studies in archosaurs. Notably, we demonstrate that the pattern of endocranial shape variation closely reflects brain shape variation. Therefore, analyses of endocranial morphology are unlikely to generate spurious conclusions about large‐scale trends in brain size and shape. To mitigate any artifacts, however, paleoneurological studies should consider the lower brain–endocast correspondence in the hindbrain relative to the forebrain; higher size and shape correspondences in chickens than alligators throughout postnatal ontogeny; artificially ‘pedomorphic’ shape of endocasts relative to their corresponding brains; and potential biases in both size and shape data due to the lack of control for ontogenetic stages in endocranial sampling.

A Step-by-Step Guide for Geometric Morphometrics of Floral Symmetry.

This paper provides a step-by-step guide for the morphological analysis of corolla and the decomposition of corolla shape variation into its symmetric and asymmetric components. The shape and symmetric organisation of corolla are key traits in the developmental and evolutionary biology of flowering plants. The various spatial layout of petals can exhibit bilateral symmetry, rotational symmetry or more complex combination of symmetry types. Here, I describe a general landmark-based geometric morphometric framework for the full statistical shape analysis of corolla and exemplify its use with four fully worked out case studies including tissue treatment, imaging, landmark data collection, file formatting, and statistical analyses: (i) bilateral symmetry (Fedia graciliflora), (ii) two perpendicular axes of bilateral symmetry (Erysimum mediohispanicum), (iii) rotational symmetry (Vinca minor), and (iv) combined bilateral and rotational symmetry (Trillium undulatum). The necessary tools for such analyses are not implemented in standard morphometric software and they are therefore provided here as functions running in the R environment. Principal Component Analysis is used to separate symmetric and asymmetric components of variation, respectively, quantifying variation among and within individuals. For bilaterally symmetric flowers, only one component of left–right asymmetric variation is extracted, while flowers with more complex symmetric layout have components of asymmetric variation associated with each symmetry operator implied (e.g., left–right asymmetry and adaxial–abaxial asymmetry). Fundamental information on the genetic, developmental, and environmental determinants of shape variation can be inferred from this decomposition (e.g., directional asymmetry, fluctuating asymmetry) and further exploited to document patterns of canalization, developmental stability, developmental modularity and morphological integration. Even if symmetry and asymmetry are not the primary interest of a study on corolla shape variation, statistical and anatomical arguments support the use of the framework advocated. This didactic protocol will help both morphometricians and non-morphometricians to further understand the role of symmetry in the development, variation and adaptive evolution of flowers.

Ecomorphology of a generalist freshwater gastropod: complex relations of shell morphology, habitat, and fecundity

Evolutionary and ecological situations in a species’ native and invasive ranges can be drastically different. This is the case for Potamopyrgus antipodarum Gray (1843) a morphologically highly variable freshwater snail native to New Zealand, where sexual and asexual individuals coexist and experience selective pressure by sterilizing endoparasites. By contrast, only a few asexual lineages have been established in invaded regions around the globe, where parasite infection is extremely rare. We analyzed the ecomorphology of 996 native P. antipodarum in a geometric morphometric framework, using brood size as proxy for fecundity, and mtDNA and nuclear SNPs to account for relatedness and identify reproductive mode. As expected, we found genetic and morphological diversity to be higher in native than in invasive snails investigated previously, but surprisingly no higher morphological diversity in sexual versus asexual individuals. The relationships between shell morphology, habitat, and fecundity were complex. Shape variation was primarily linked to genetic relatedness but specific environmental factors including flow rate induced similar shell shapes. By contrast, shell size was largely explained by environmental factors. Fecundity was correlated with size but showed trade-offs with shape in increasingly extreme conditions. With increasing flow and toward small springs, the trend of shell shape becoming wider was reversed, i.e., snails with narrower shells were brooding more embryos. We concluded that both genetic and environmental contributions to variation in shell morphology in P. antipodarum likely play an important role in the ability of this species to adapt to a wide spectrum of habitats.

Integrative taxonomy of Afrotropical Ornithodoros (Ornithodoros) (Acari: Ixodida: Argasidae)

Afrotropical Ornithodoros (Ornithodoros) ticks are revised based on qualitative morphology of females and nymphs, as well as tarsus I shape outlines of females measured in a geometric morphometric framework. These lines of evidence corroborate lineages based on 16S rRNA nucleotide sequence data. Four previously unrecognized species are described, along with a revived nomen nudum that was previously considered a synonym. Afrotropical Ornithodoros (Ornithodoros) now comprise ten species. Ornithodoros moubata and Ornithodoros porcinus are separated from three other species in southern Africa (Ornithodoros compactus, Ornithodoros phacochoerusn. sp., Ornithodoros waterbergensisn. sp.), with O. porcinus restricted to central east Africa. Known species boundaries for Ornithodoros apertus and O. compactus are supported. Ornithodoros savignyi are separated from three other species in South Africa and Namibia, with O. savignyi restricted to north Africa. Neumann’s Ornithodoros pavimentosusnom. rev. are resurrected from synonymy as a species that occur in Bushmanland, Namaqualand and Namibia, while Ornithodoros kalahariensisn. sp. occur in Kalahari thornveld, and Ornithodoros noorsveldensisn. sp. occur in Noorsveld thicket of South Africa. Detailed descriptions are given for each species along with high resolution images and point map distributions. Support is provided for speciation driven by riverine barriers, Pliocene uplift and differential arid tolerance. Exaggerated tarsus I shape in the O. savignyi group suggests adaptation to fossorial habits and soil type. Conversely, reduced tarsus I shape in the O. moubata group is suggested as an evolutionary consequence of the life history change from soil to warthog burrows. This study represents an integrative (iterative) approach to delimiting Afrotropical Ornithodoros (Ornithodoros) species, and provides the first application of tarsus I shape outlines in a geometric morphometric framework for testing species boundaries.

A shape distance based on the Fisher–Rao metric and its application for shapes clustering

In the clustering of shapes, which is a longstanding challenge in the framework of geometric morphometrics and shape analysis, is crucial the selection and application of a suitable and appropriate measurement of distance among observations (i.e. individuals). The aim of this study is to model shapes from complex systems using Information Geometry tools.It is well-known that the Fisher information endows the statistical manifold, defined by a family of probability distributions, with a Riemannian metric, called the Fisher–Rao metric. With respect to this, geodesic paths are determined, minimizing information in Fisher sense. The geodesic distance induced by the Fisher–Rao metric can be used to define a shape metric which enables us to quantify differences between shapes.The discriminative power of the proposed Fisher–Rao distance is tested in the context of shapes clustering on both simulated and real data sets. Results show a better ability in recovering the true cluster structure with respect to the standard Kendall’s shape metric.

Investigating Morphospace Occupation in Multi-Scale Ecological and Evolutionary Data Using Regression Tree: Case Studies and Perspectives

A key challenge in ecology and evolutionary biology is to explain the origin, structure and temporal patterns of phenotypic diversity. With regard to the potentially complex determinism of phenotypic differences, the issue should be comprehended in a general view, across multiple scales and an increasing number of phenomic studies investigate shape variation through large taxonomic, biogeographic or temporal scales. In this context, there is an ever-increasing need to develop new tools for a coherent understanding of morphospace occupation by disentangling and quantifying the main determinants of phenotypic changes. The present study briefly introduce the possibility to use multivariate regression tree technique to cope with morphological data, as embedded in a geometric morphometric framework. It emphasizes that hierarchical partitioning methods produce a hierarchy between causal variables that may help analyzing complexity in multi-scale ecological and evolutionary data. I therefore suggest that morphological studies would benefit from the combined use of the classical statistical models with rapidly emerging and diversifying methods of machine-learning. Doing so allows one to primary explore in an extensive exploratory manner the hierarchy of nested organisational levels underlying morphological variation, and then conduct hypothesis-driven analysis by focusing on a relevant scale or by investigating the appropriate model that reflects hypothesized nested influence of explanatory variables. The outlined approach may help investigating morphospace occupation in an explicitly hierarchical quantitative context.

Quantification of Osteon Morphology Using Geometric Histomorphometrics.

Many histological methods in forensic anthropology utilize combinations of traditional histomorphometric parameters which may not accurately describe the morphology of microstructural features. Here, we report the novel application of a geometric morphometric method suitable when considering structures without anatomically homologous landmarks for the quantification of complete secondary osteon size and morphology. The method is tested for its suitability in the measurement of intact secondary osteons using osteons digitized from transverse femoral diaphyseal sections prepared from two human individuals. The results of methodological testing demonstrate the efficacy of the technique when applied to intact secondary osteons. In providing accurate characterization of micromorphology within the robust mathematical framework of geometric morphometrics, this method may surpass traditional histomorphometric variables currently employed in forensic research and practice. A preliminary study of the intersectional histomorphometric variation within the femoral diaphysis is made using this geometric histomorphometric method to demonstrate its potential.

Constant fluctuating asymmetry but not directional asymmetry along the geographic distribution of Drosophila antonietae (Diptera, Drosophilidae)

The population dynamics of a species tends to change from the core to the periphery of its distribution. Therefore, one could expect peripheral populations to be subject to a higher level of stress than more central populations (the center–periphery hypothesis) and consequently should present a higher level of fluctuating asymmetry. To test these predictions we study asymmetry in wing shape of five populations of Drosophila antonietae collected throughout the distribution of the species using fluctuating asymmetry as a proxy for developmental instability. More specifically, we addressed the following questions: (1) what types of asymmetry occur in populations of D. antonietae? (2) Does the level of fluctuating asymmetry vary among populations? (3) Does peripheral populations have a higher fluctuating asymmetry level than central populations? We used 12 anatomical landmarks to quantify patterns of asymmetry in wing shape in five populations of D. antonietae within the framework of geometric morphometrics. Net asymmetry – a composite measure of directional asymmetry+fluctuating asymmetry – varied significantly among populations. However, once net asymmetry of each population is decomposed into directional asymmetry and fluctuating asymmetry, most of the variation in asymmetry was explained by directional asymmetry alone, suggesting that populations of D. antonietae have the same magnitude of fluctuating asymmetry throughout the geographical distribution of the species. We hypothesize that larval development in rotting cladodes might play an important role in explaining our results. In addition, our study underscores the importance of understanding the interplay between the biology of a species and its geographical patterns of asymmetry.

Disentangling and quantifying sources of otolith shape variation across multiple scales using a new hierarchical partitioning approach

Otolith shape analysis provides a practical basis for stock separation, useful in fisheries management. A wide range of factors affect otolith morphology and a fundamental challenge in morphometric stock identification is to develop a consensus on the biological interpretation of otolith shape variation. To date, there are few comprehensive reports that have examined various factors simultaneously at multiple scales in natural conditions. In this study, I identified sources of otolith shape variation across spatial and taxonomic scales using a new hierarchical partitioning method embedded in a geometric morphometric framework. Various environmental, taxonomic and endogenic factors which affect otolith shape were quantitatively investigated in 2077 coral reef fishes. Using the new partitioning method, allometry accounted for a considerable degree of otolith shape variation at all scales and contributed more variation to regional differences than did habitats or islands. While large-scale variations are expected to be associated with significant shape variation, the study provides quantitative evidence that both local environmental variables and large-scale patterns contribute equally to total otolith shape variation. Models that aim at discriminating stocks and forecasting stock boundaries implicitly assume that the within-stock morphological variation is negligible in regard to the variation that exists among stocks. The importance of local environmental variables may therefore act as an important confounding effect into those predictive models. More generally, the degree of contribution of some variables differed substantially among the taxonomic scales, as did their relationship among spatial scales. A scale dependent understanding of factors affecting otolith morphology is critical in understanding theintegrity of fish populations and is an effective tool in management of fisheries resources

The morphometrics of "masculinity" in human faces.

In studies of social inference and human mate preference, a wide but inconsistent array of tools for computing facial masculinity has been devised. Several of these approaches implicitly assumed that the individual expression of sexually dimorphic shape features, which we refer to as maleness, resembles facial shape features perceived as masculine. We outline a morphometric strategy for estimating separately the face shape patterns that underlie perceived masculinity and maleness, and for computing individual scores for these shape patterns. We further show how faces with different degrees of masculinity or maleness can be constructed in a geometric morphometric framework. In an application of these methods to a set of human facial photographs, we found that shape features typically perceived as masculine are wide faces with a wide inter-orbital distance, a wide nose, thin lips, and a large and massive lower face. The individual expressions of this combination of shape features—the masculinity shape scores—were the best predictor of rated masculinity among the compared methods (r = 0.5). The shape features perceived as masculine only partly resembled the average face shape difference between males and females (sexual dimorphism). Discriminant functions and Procrustes distances to the female mean shape were poor predictors of perceived masculinity.

An investigation of matching symmetry in the human pinnae with possible implications for 3D ear recognition and sound localization.

The human external ears, or pinnae, have an intriguing shape and, like most parts of the human external body, bilateral symmetry is observed between left and right. It is a well-known part of our auditory sensory system and mediates the spatial localization of incoming sounds in 3D from monaural cues due to its shape-specific filtering as well as binaural cues due to the paired bilateral locations of the left and right ears. Another less broadly appreciated aspect of the human pinna shape is its uniqueness from one individual to another, which is on the level of what is seen in fingerprints and facial features. This makes pinnae very useful in human identification, which is of great interest in biometrics and forensics. Anatomically, the type of symmetry observed is known as matching symmetry, with structures present as separate mirror copies on both sides of the body, and in this work we report the first such investigation of the human pinna in 3D. Within the framework of geometric morphometrics, we started by partitioning ear shape, represented in a spatially dense way, into patterns of symmetry and asymmetry, following a two-factor anova design. Matching symmetry was measured in all substructures of the pinna anatomy. However, substructures that 'stick out' such as the helix, tragus, and lobule also contained a fair degree of asymmetry. In contrast, substructures such as the conchae, antitragus, and antihelix expressed relatively stronger degrees of symmetric variation in relation to their levels of asymmetry. Insights gained from this study were injected into an accompanying identification setup exploiting matching symmetry where improved performance is demonstrated. Finally, possible implications of the results in the context of ear recognition as well as sound localization are discussed.

Amplified fragment length polymorphisms, the evolution of the land snail genusTheba(Stylommatophora: Helicidae), and an objective approach for relating fossils to internal nodes of a phylogenetic tree using geometric morphometrics

Time-trees are commonly calibrated based on fossils attributed to internal nodes, thus defining the minimum age of these nodes. However, in the absence of synapomorphies, the phylogenetic position of a fossil can only be inferred based on similarity. In this study, we objectively allocated fossil shells to internal nodes based on the reconstruction of ancestral shapes and sizes in a geometric morphometric framework. Our phylogenetic analysis of 24 (putative sub)species was based on 2524 amplified fragment length polymorphism loci. In this well-supported tree the taxa occurring in north-west Africa and on the Iberian peninsula were paraphyletic with respect to the (sub)species from the Canary and Selvagen Islands, indicating a continental origin of the genus in contrast to our earlier sequence-based account. Ancestral shell shapes and sizes were inferred based on landmark data using squared-change parsimony. In a subsequent principal component analysis, only three of 20 fossil shells could be unequivocally allocated to internal nodes. However, these fossils were all Quaternary, and thus too young to infer meaningfully narrow confidence intervals for divergence estimates that probably reach back into the Miocene or even Oligocene. The apparent failure of allocating older fossils to internal nodes and achieving a reliable tree calibration was caused by the absence of phylogenetic signal in the shells of Theba due to extensive, sometimes rapid, convergent evolution, including reversals. © 2014 The Linnean Society of London

Salinity and Egg Shape Variation: A Geometric Morphometric Analysis

The harvest and incubation of American Alligator (Alligator mississippiensis) eggs is an important component to the commercial alligator harvest industry in the southeastern United States. As a result, various methodologies have been used to monitor alligator populations including abundance counts, stress quantification, and nesting surveys. Past studies have dismissed the importance of egg shape in crocodilians, Squamates, and turtles and deemed egg shape in birds and other amniotes as similar, in relation to functionality. The complexity of crocodilian eggs has been examined, and both turtle and Squamate eggs have been regarded recently as physiologically more intricate than bird eggs. This study introduces a physiological approach to monitor alligator populations in freshwater and low salinity environments by quantifying egg shape in correlation with varying salinity. We introduce a fractional semilandmark-shape template method to quantify egg shape within a geometric morphometric framework. This approach is beneficial because it allows for the quantification of shape for curved structures, such as eggs, which lack homologous landmarks. The results from this study suggest that alligator egg shape is correlated with varying salinity levels, such that variation in alligator egg shape at low salinities changes in gradient-like fashion, whereas salinities high enough to be deemed stressful result in reversion back to a low salinity egg shape or desiccation. This study elucidates a correlation that can be implemented in management and breeding techniques and opens the door to in-depth physiological examination of the system.

Sexual dimorphism in multiple aspects of 3D facial symmetry and asymmetry defined by spatially dense geometric morphometrics

Accurate measurement of facial sexual dimorphism is useful to understanding facial anatomy and specifically how faces influence, and have been influenced by, sexual selection. An important facial aspect is the display of bilateral symmetry, invoking the need to investigate aspects of symmetry and asymmetry separately when examining facial shape. Previous studies typically employed landmarks that provided only a sparse facial representation, where different landmark choices could lead to contrasting outcomes. Furthermore, sexual dimorphism is only tested as a difference of sample means, which is statistically the same as a difference in population location only. Within the framework of geometric morphometrics, we partition facial shape, represented in a spatially dense way, into patterns of symmetry and asymmetry, following a two-factor anova design. Subsequently, we investigate sexual dimorphism in symmetry and asymmetry patterns separately, and on multiple aspects, by examining (i) population location differences as well as differences in population variance-covariance; (ii) scale; and (iii) orientation. One important challenge in this approach is the proportionally high number of variables to observations necessitating the implementation of permutational and computationally feasible statistics. In a sample of gender-matched young adults (18-25 years) with self-reported European ancestry, we found greater variation in male faces than in women for all measurements. Statistically significant sexual dimorphism was found for the aspect of location in both symmetry and asymmetry (directional asymmetry), for the aspect of scale only in asymmetry (magnitude of fluctuating asymmetry) and, in contrast, for the aspect of orientation only in symmetry. Interesting interplays with hypotheses in evolutionary and developmental biology were observed, such as the selective nature of the force underpinning sexual dimorphism and the genetic independence of the structural patterns of fluctuating asymmetry. Additionally, insights into growth patterns of the soft tissue envelope of the face and underlying skull structure can also be obtained from the results.

Genetic structure and phenotypic diversity of two northern populations of Cheilosia aff. longula (Diptera: Syrphidae) has implications for evolution and conservation

The genetic structure and phenotypic diversity of two populations of Cheilosia aff. longula (Diptera: Syrphidae) in Lapland, Finland, were examined using DNA sequencing, protein electrophoresis, and geometric morphometrics. The morphological identification of the species were verified using partial sequences of mitochondrial cytochrome c oxidase subunit I (COI mtDNA), and the nuclear ribosomal internal transcribed region 2 (ITS2 rDNA), and comparing the corresponding sequences of Cheilosia aff. longula and the closely related C. longula. Two and three haplotypes of the genes COI mtDNA and ITS2 rDNA were identified in the two populations. Analysis of 12 isozyme loci showed an extremely low genetic variability in the populations originating from Utsjoki and Kevo. Discriminant analysis combined with canonical variate analysis revealed inter-population divergence in wing shape. Variation among genetically diverse individuals, both within- and among studied populations was studied, and directional (DA) and fluctuating asymmetry (FA) estimated using landmarks in the framework of geometric morphometrics. It is likely that the documented DA and FA asymmetry in both wing shape and size reflects the developmental instability of the individuals studied. By using Procrustes ANOVA the locations of particular landmarks responsible for the variation in shape were determined. The decomposition of the components of variance accorded to each landmark showed that the landmarks differed in the percentage of variation they accounted for (DA, FA and variation among individuals). In the discussion the implications of the reduced genetic diversity and asymmetry in wing traits for conservation is considered.