The Chalcidoidea bush of life: evolutionary history of a massive radiation of minute wasps.

Chalcidoidea are mostly parasitoid wasps that include as many as 500 000 estimated species. Capturing phylogenetic signal from such a massive radiation can be daunting. Chalcidoidea is an excellent example of a hyperdiverse group that has remained recalcitrant to phylogenetic resolution. We combined 1007 exons obtained with Anchored Hybrid Enrichment with 1048 ultra-conserved elements (UCEs) for 433 taxa including all extant families, >95% of all subfamilies, and 356 genera chosen to represent the vast diversity of the superfamily. Going back and forth between the molecular results and our collective knowledge of morphology and biology, we detected bias in the analyses that was driven by the saturation of nucleotide data. Our final results are based on a concatenated analysis of the least saturated exons and UCE datasets (2054 loci, 284 106 sites). Our analyses support an expected sister relationship with Mymarommatoidea. Seven previously recognized families were not monophyletic, so support for a new classification is discussed. Natural history in some cases would appear to be more informative than morphology, as illustrated by the elucidation of a clade of plant gall associates and a clade of taxa with planidial first-instar larvae. The phylogeny suggests a transition from smaller soft-bodied wasps to larger and more heavily sclerotized wasps, with egg parasitism as potentially ancestral for the entire superfamily. Deep divergences in Chalcidoidea coincide with an increase in insect families in the fossil record, and an early shift to phytophagy corresponds with the beginning of the "Angiosperm Terrestrial Revolution". Our dating analyses suggest a middle Jurassic origin of 174 Ma (167.3-180.5 Ma) and a crown age of 162.2 Ma (153.9-169.8 Ma) for Chalcidoidea. During the Cretaceous, Chalcidoidea may have undergone a rapid radiation in southern Gondwana with subsequent dispersals to the Northern Hemisphere. This scenario is discussed with regard to knowledge about the host taxa of chalcid wasps, their fossil record and Earth's palaeogeographic history.

Classifying fossil Darwin wasps (Hymenoptera: Ichneumonidae) with geometric morphometrics of fore wings.

Linking fossil species to the extant diversity is often a difficult task, and the correct interpretation of character evidence is crucial for assessing their taxonomic placement. Here, we make use of geometric morphometrics of fore wings to help classify five fossil Darwin wasps from the Early Eocene Fur Formation in Denmark into subfamilies and often tribes. We compile a reference dataset with 342 fore wings of nine extant subfamilies and nine relevant fossil species. Since geometric morphometrics was mostly ignored in the past in Darwin wasp classification, the dataset is first used to examine differences and similarities in wing venation among subfamilies. In a next step, we used the reference dataset to inform the classification of the fossil species, which resulted in the description of one new genus and five new species, Crusopimpla weltii sp. nov., Ebriosa flava gen. et sp. nov., Entypoma? duergari sp. nov., Lathrolestes? zlatorog sp. nov., and Triclistus bibori sp. nov., in four different subfamilies. Carefully assessing data quality, we show that the fore wing venation of fossil Darwin wasps is surprisingly suitable to assign them to a subfamily or even lower taxonomic level, especially when used in conjunction with characters from other parts of the body to narrow down a candidate set of potential subfamilies and tribes. Our results not only demonstrate a fast and useful approach to inform fossil classification but provide a basis for future investigations into evolutionary changes in fore wings of ichneumonids. The high informativeness of wing venation for classification furthermore could be harvested for phylogenetic analyses, which are otherwise often hampered by homoplasy in this parasitoid wasp family.

Tiny wasps, huge diversity - A review of German Pteromalidae with new generic and species records (Hymenoptera: Chalcidoidea).

BACKGROUND: Despite their ecological and economic importance, hymenopteran parasitoids are severely understudied. Even in countries with a long taxonomic history such as Germany, dating back to the 18th century and including prolific figures like Christian Gottfired Nees von Esenbeck and Otto Schmiedeknecht, those species-rich groups are seldom the subject of comprehensive research efforts, leaving their true diversity unknown. This is often due to their small size of a few millimetres on average, leading to difficulties in their identification and examination. The chalcidoid family Pteromalidae is no exception to this neglect. So far, 735 species have been reported from Germany. Estimating the diversity of this group is not possible, but it has to be assumed that many more species are still to be discovered in Germany. NEW INFORMATION: With this study, we improve the knowledge on pteromalid diversity and present new records of 17 genera and 41 species, previously unknown to occur in Germany. We also match and describe previously unknown sexes of two species, based on DNA barcode data. The results of this study were generated as part of the German Barcode of Life Project. The newly-recorded species are illustrated and notes on the biology and distribution are given. The ecological significance of Pteromalidae and potential value as indicators for nature conservation efforts are briefly discussed.

Two species? - Limits of the species concepts in the pygmy grasshoppers of the Tetrix bipunctata complex (Orthoptera, Tetrigidae).

Today, integrative taxonomy is often considered the gold standard when it comes to species recognition and delimitation. Using the Tetrix bipunctata complex, we here present a case where even integrative taxonomy may reach its limits. The Tetrix bipunctata complex consists of two morphs, bipunctata and kraussi, which are easily distinguished by a single character, the length of the hind wing. Both morphs are widely distributed in Europe and reported to occur over a large area in sympatry, where they occasionally may live also in syntopy. The pattern has led to disparate classifications, as on the one extreme, the morphs were treated merely as forms or subspecies of a single species, on the other, as separate species. For this paper, we re-visited the morphology by using multivariate ratio analysis (MRA) of 17 distance measurements, checked the distributional data based on verified specimens and examined micro-habitat use. We were able to confirm that hind wing length is, indeed, the only morphological difference between bipunctata and kraussi. We were also able to exclude a mere allometric scaling. The morphs are, furthermore, largely sympatrically distributed, with syntopy occurring regularly. However, a microhabitat niche difference can be observed. Ecological measurements in a shared habitat confirm that kraussi prefers a drier and hotter microhabitat, which possibly also explains the generally lower altitudinal distribution. Based on these results, we can exclude classification as subspecies, but the taxonomic classification as species remains unclear. Even with different approaches to classify the Tetrix bipunctata complex, this case is, therefore, not settled. We recommend continuing to record kraussi and bipunctata separately.

Incongruent molecular and morphological variation in the crab spider Synemaglobosum (Araneae, Thomisidae) in Europe.

Establishing species boundaries is one of the challenges taxonomists around the world have been tackling for centuries. The relation between intraspecific and interspecific variability is still under discussion and in many taxa it remains understudied. Here the hypothesis of single versus multiple species of the crab spider Synemaglobosum (Fabricius) is tested. The wide distribution range as well as its high morphological variability makes this species an interesting candidate for re-evaluation using an integrative approach. This study combines information from barcoding, phylogenetic reconstruction based on mitochondrial CO1 and ITS2 of more than 60 specimens collected over a wide range of European localities, and morphology. The findings show deep clades with up to 6% mean pairwise distance in the CO1 barcode without any biogeographical pattern. The nuclear ITS2 gene did not support the CO1 clades. Morphological assessment of somatic and genital characters in males and females and a morphometric analysis of the male palp uncovered high intraspecific variation that does not match the CO1 or ITS2 phylogenies or biogeography either. Screening for endosymbiotic Wolbachia bacteria was conducted and only a single infected specimen was found. Several scenarios might explain these inconsistent patterns. While the deep divergences in the barcoding marker might suggest cryptic or ongoing speciation or geographical isolation in the past, the lack of congruent variation in the nuclear ITS2 gene or the studied morphological character systems, especially the male palp, indicates that S.globosum might simply be highly polymorphic both in terms of its mtDNA and morphology. Therefore, more data on ecology and behaviour and full genome sequences are necessary to ultimately resolve this taxonomically intriguing case.

Cuticular hydrocarbons as potential mediators of cryptic species divergence in a mutualistic ant association.

Upon advances in sequencing techniques, more and more morphologically identical organisms are identified as cryptic species. Often, mutualistic interactions are proposed as drivers of diversification. Species of the neotropical parabiotic ant association between Crematogaster levior and Camponotus femoratus are known for highly diverse cuticular hydrocarbon (CHC) profiles, which in insects serve as desiccation barrier but also as communication cues. In the present study, we investigated the association of the ants' CHC profiles with genotypes and morphological traits, and discovered cryptic species pairs in both genera. To assess putative niche differentiation between the cryptic species, we conducted an environmental association study that included various climate variables, canopy cover, and mutualistic plant species. Although mostly sympatric, the two Camponotus species seem to prefer different climate niches. However in the two Crematogaster species, we could not detect any differences in niche preference. The strong differentiation in the CHC profiles may thus suggest a possible role during speciation itself either by inducing assortative mating or by reinforcing sexual selection after the speciation event. We did not detect any further niche differences in the environmental parameters tested. Thus, it remains open how the cryptic species avoid competitive exclusion, with scope for further investigations.

Larval and phenological traits predict insect community response to mowing regime manipulations.

For the restoration of biodiversity in agricultural grasslands, it is essential to understand how management acts as an ecological filter on the resident species. Mowing constitutes such a filter: only species that possess functional traits enabling them to withstand its consequences can persist in the community. We investigated how the timing of mowing modulates this filtering effect for insects. We predicted that two traits drive species responses. Species with larval development within the meadow vegetation will suffer more from mowing than species whose larvae develop in or on the ground, or outside the meadows, while species with a later phenology should benefit from later mowing. We conducted a five-year experiment, replicated at 12 sites across the Swiss lowlands, applying three different mowing regimes to low-intensity hay meadows: (1) first cut of the year not earlier than 15 June (control regime); (2) the first cut delayed until 15 July; and (3) leaving an uncut grass refuge on 10-20% of the meadow area (after earliest first cut on 15 June). Before the first cut in years 4 or 5, we sampled larvae of Lepidoptera and sawflies, and adults of moths, parasitoid wasps, wild bees, hoverflies, ground beetles, and rove beetles. Overall, before the first cut of the year, abundances of species with vegetation-dwelling larvae were higher in meadows with delayed mowing or an uncut grass refuge, with some taxon-specific variation. In contrast, species whose larval development is independent of the meadow vegetation showed no differences in abundance between mowing regimes. Species richness did not differ among regimes. For species with vegetation-dwelling larvae, a fourth-corner analysis showed an association between early phenology and the control regime. No associations were found for the other functional groups. Our results show that slight modifications of mowing regimes, easily implementable in agri-environmental policy schemes, can boost invertebrate abundance, potentially benefitting insectivorous vertebrates.

An interactive key to Central European species of the Pteromalus albipennis species group and other species of the genus (Hymenoptera: Chalcidoidea: Pteromalidae), with the description of a new species.

BACKGROUND: Parasitoid wasps of the genus Pteromalus play an important role in biological pest control, however, the genus includes a large number of cryptic species, which makes reliable identification difficult. The latest identification key dates back to Graham (1969) and since then many new species have been described and nomenclatural changes proposed. NEW INFORMATION: Here we present an interactive and fully illustrated identification key in Xper3 for 27 species of the Pteromalus albipennis species group as well as for 18 similar species. In addition to qualitative traits, a large set of body measurements is incorporated in the key. We also explored a new set of qualitative features on the propodeum and metasternum. During field work, a new species of the P. albipennis species group, P. capito Baur sp. n., could be reared from flower heads of Asteraceae, which is described here. It looks very similar to P. albipennis and P. cingulipes, however, several qualitative characters and body ratios distinguish it clearly from the most similar species.

Plant diversity has contrasting effects on herbivore and parasitoid abundance in Centaurea jacea flower heads.

High biodiversity is known to increase many ecosystem functions, but studies investigating biodiversity effects have more rarely looked at multi-trophic interactions. We studied a tri-trophic system composed of Centaurea jacea (brown knapweed), its flower head-infesting tephritid fruit flies and their hymenopteran parasitoids, in a grassland biodiversity experiment. We aimed to disentangle the importance of direct effects of plant diversity (through changes in apparency and resource availability) from indirect effects (mediated by host plant quality and performance). To do this, we compared insect communities in C. jacea transplants, whose growth was influenced by the surrounding plant communities (and where direct and indirect effects can occur), with potted C. jacea plants, which do not compete with the surrounding plant community (and where only direct effects are possible). Tephritid infestation rate and insect load, mainly of the dominant species Chaetorellia jaceae, decreased with increasing plant species and functional group richness. These effects were not seen in the potted plants and are therefore likely to be mediated by changes in host plant performance and quality. Parasitism rates, mainly of the abundant chalcid wasps Eurytoma compressa and Pteromalus albipennis, increased with plant species or functional group richness in both transplants and potted plants, suggesting that direct effects of plant diversity are most important. The differential effects in transplants and potted plants emphasize the importance of plant-mediated direct and indirect effects for trophic interactions at the community level. The findings also show how plant-plant interactions critically affect results obtained using transplants. More generally, our results indicate that plant biodiversity affects the abundance of higher trophic levels through a variety of different mechanisms.

Description of Brachymeria philornisae sp. n. (Hymenoptera: Chalcididae), a parasitoid of the bird parasite Philornis trinitensis (Diptera: Muscidae) in Tobago, with a review of the sibling species.

In this paper, we describe and illustrate a new species of parasitoid wasp as Brachymeria philornisae Delvare, sp. nov. The new species was reared from Philornis trinitensis Dodge & Aitken (Diptera: Muscidae) puparia that were found in the nests of the bird species Mimus gilvus (Vieillot) (Mimidae) and Tiaris bicolor (L.) (Thraupidae) in Tobago. The new species is of particular interest as it may be considered a potential biological control agent in locations where Philornis species are invasive, such as the Galapagos Islands. Closely related Brachymeria species had taxonomically ambiguous relationship in the past and are compared and reviewed. The species have been classified in the subgenus Pseudobrachymeria, but are here treated within the newly defined subconica species-group of Brachymeria as part of a sibling species complex designated as the subrugosa complex. Species assigned to the subconica species-group are listed and five, one unnamed, are assigned to the subrugosa complex and their females keyed. Species are separated by qualitative characters and morphometry using distance measurements. Trigonura annulipes Costa Lima is renamed as Brachymeria costalimai Delvare nom. nov. because of secondary homonymy.

Complex within a Complex: Integrative Taxonomy Reveals Hidden Diversity in Cicadetta brevipennis (Hemiptera: Cicadidae) and Unexpected Relationships with a Song Divergent Relative.

Multiple sources of data in combination are essential for species delimitation and classification of difficult taxonomic groups. Here we investigate a cicada taxon with unusual cryptic diversity and we attempt to resolve seemingly contradictory data sets. Cicada songs act as species-specific premating barriers and have been used extensively to reveal hidden taxonomic diversity in morphologically similar species. The Palaearctic Cicadetta montana species complex is an excellent example where distinct song patterns have disclosed multiple recently described species. Indeed, two taxa turned out to be especially diverse in that they form a "complex within the complex": the Cicadetta cerdaniensis song group (four species studied previously) and Cicadetta brevipennis (examined in details here). Based on acoustic, morphological, molecular, ecological and spatial data sampled throughout their broad European distribution, we find that Cicadetta brevipennis s. l. comprises five lineages. The most distinct lineage is identified as Cicadetta petryi Schumacher, 1924, which we re-assign to the species level. Cicadetta brevipennis litoralis Puissant & Hertach ssp. n. and Cicadetta brevipennis hippolaidica Hertach ssp. n. are new to science. The latter hybridizes with Cicadetta brevipennis brevipennis Fieber, 1876 at a zone inferred from intermediate song patterns. The fifth lineage requires additional investigation. The C. cerdaniensis and the C. brevipennis song groups exhibit characteristic, clearly distinct basic song patterns that act as reproductive barriers. However, they remain completely intermixed in the Bayesian and maximum likelihood COI and COII mitochondrial DNA phylogenies. The closest relative of each of the four cerdaniensis group species is a brevipennis group taxon. In our favoured scenario the phylogenetic pairs originated in common Pleistocene glacial refuges where the taxa speciated and experienced sporadic inter-group hybridization leading to extensive introgression and mitochondrial capture.

Striking cuticular hydrocarbon dimorphism in the mason wasp Odynerus spinipes and its possible evolutionary cause (Hymenoptera: Chrysididae, Vespidae).

Cleptoparasitic wasps and bees smuggle their eggs into the nest of a host organism. Here the larvae of the cleptoparasite feed upon the food provision intended for the offspring of the host. As cleptoparasitism incurs a loss of fitness for the host organism (offspring of the host fail to develop), hosts of cleptoparasites are expected to exploit cues that alert them to potential cleptoparasite infestation. Cuticular hydrocarbons (CHCs) could serve as such cues, as insects inevitably leave traces of them behind when entering a nest. By mimicking the host's CHC profile, cleptoparasites can conceal their presence and evade detection by their host. Previous studies have provided evidence of cleptoparasites mimicking their host's CHC profile. However, the impact of this strategy on the evolution of the host's CHC profile has remained unexplored. Here, we present results from our investigation of a host-cleptoparasite system consisting of a single mason wasp species that serves syntopically as the host to three cuckoo wasp species. We found that the spiny mason wasp (Odynerus spinipes) is able to express two substantially different CHC profiles, each of which is seemingly mimicked by a cleptoparasitic cuckoo wasp (i.e. Chrysis mediata and Pseudospinolia neglecta). The CHC profile of the third cuckoo wasp (Chrysis viridula), a species not expected to benefit from mimicking its host's CHC profile because of its particular oviposition strategy, differs from the two CHC profiles of its host. Our results corroborate the idea that the similarity of the CHC profiles between cleptoparasitic cuckoo wasps and their hosts are the result of chemical mimicry. They further suggest that cleptoparasites may represent a hitherto unappreciated force that drives the evolution of their hosts' CHCs.

Pushing the limits - two new species of Pteromalus (Hymenoptera, Chalcidoidea, Pteromalidae) from Central Europe with remarkable morphology.

Two new species, Pteromalusbriani sp. n. and Pteromalusjanstai sp. n., with unusual characters are described from the Central Plateau and the Alps in Switzerland, respectively. Pteromalusbriani sp. n. is remarkable in that it has the metatibia quite abruptly expanded before the middle. This type of modification of the hind tibia is unique within the Pteromalidae and probably also the entire Chalcidoidea. It is also very rare in other parasitic wasps, where it is suspected to be associated with pheromone glands. The species is a gregarious endoparasitoid of pupae of Vanessaatalanta (Linnaeus) and Aglaisurticae (Linnaeus), two common butterflies (Lepidoptera: Nymphalidae) in Europe. It is furthermore a koinobiont parasitoid ovipositing in an early larval stage of the host. The other species, Pteromalusjanstai sp. n., shows a flattened mesosoma. A dorsoventrally depressed body is a unique feature within the genus Pteromalus, but known from a number species in unrelated genera and subfamilies. The two records demonstrate that it is possible to discover entirely new species with extraordinary characters even in one of the taxonomically most thoroughly explored parts of the world.

Does early learning drive ecological divergence during speciation processes in parasitoid wasps?

Central to the concept of ecological speciation is the evolution of ecotypes, i.e. groups of individuals occupying different ecological niches. However, the mechanisms behind the first step of separation, the switch of individuals into new niches, are unclear. One long-standing hypothesis, which was proposed for insects but never tested, is that early learning causes new ecological preferences, leading to a switch into a new niche within one generation. Here, we show that a host switch occurred within a parasitoid wasp, which is associated with the ability for early learning and the splitting into separate lineages during speciation. Lariophagus distinguendus consists of two genetically distinct lineages, most likely representing different species. One attacks drugstore beetle larvae (Stegobium paniceum (L.)), which were probably the ancestral host of both lineages. The drugstore beetle lineage has an innate host preference that cannot be altered by experience. In contrast, the second lineage is found on Sitophilus weevils as hosts and changes its preference by early learning. We conclude that a host switch has occurred in the ancestor of the second lineage, which must have been enabled by early learning. Because early learning is widespread in insects, it might have facilitated ecological divergence and associated speciation in this hyperdiverse group.

A new species of the paper wasp genus Polistes (Hymenoptera, Vespidae, Polistinae) in Europe revealed by morphometrics and molecular analyses.

We combine multivariate ratio analysis (MRA) of body measurements and analyses of mitochondrial and nuclear data to examine the status of several species of European paper wasps (Polistes Latreille, 1802) closely related to P. gallicus. Our analyses unambiguously reveal the presence of a cryptic species in Europe, as two distinct species can be recognized in what has hitherto been considered Polistes bischoffi Weyrauch, 1937. One species is almost as light coloured as P. gallicus, and is mainly recorded from Southern Europe and Western Asia. The other species is darker and has a more northern distribution in Central Europe. Both species occur syntopically in Switzerland. Given that the lost lectotype of P. bischoffi originated from Sardinia, we selected a female of the southern species as a neotype. The northern species is described as P. helveticus sp. n. here. We also provide a redescription of P. bischoffi rev. stat. and an identification key including three more closely related species, P. biglumis, P. gallicus and P. hellenicus.

Morphometric analysis and taxonomic revision of Anisopteromalus Ruschka (Hymenoptera: Chalcidoidea: Pteromalidae) - an integrative approach.

We use an integrative taxonomic approach to revise the genus Anisopteromalus. In particular, we apply multivariate ratio analysis (MRA), a rather new statistical method based on principal component analysis (PCA) and linear discriminant analysis (LDA), to numerous body measurements and combine the data with those from our molecular analysis of Cytb and ITS2 genetic markers (on a subset of species) and all available published data on morphology, karyology, behaviour, host associations and geographic distribution. We demonstrate that the analysis of quantitative characters using MRA plays a major role for the integration of name-bearing types and thus for the association of taxa with names. Six species are recognized, of which two are new: A. cornis Baur sp.n. and A. quinarius Gokhman & Baur sp.n. For Anisopteromalus calandrae (Howard), a well-known, cosmopolitan parasitoid of stored-product pests, we have selected a neotype to foster continuity and stability in the application of this important name. The species was sometimes confused with the related A. quinariussp.n., another cosmopolitan species that is frequently encountered in similar environments. We also show that several species originally described or later put under Anisopteromalus actually belong to different genera: Cyrtoptyx camerunus (Risbec) comb.n.; Meraporus glaber (Szelényi) comb.n.; Dinarmus schwenkei (Roomi, Khan & Khan) comb.n.Neocatolaccus indicus Ayyar & Mani is confirmed as a junior synonym of Oxysychus sphenopterae (Ferrière) syn.n. and Anisopteromalus calandrae brasiliensis (Domenichini) stat.rev. must be considered as a valid but doubtful taxon. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:BDFE96D3-D0F4-4012-90F5-9A087F7F5864.

A phylogenetic analysis of the megadiverse Chalcidoidea (Hymenoptera).

Chalcidoidea (Hymenoptera) is extremely diverse with an estimated 500 000 species. We present the first phylogenetic analysis of the superfamily based on both morphological and molecular data. A web-based, systematics workbench mx was used to score 945 character states illustrated by 648 figures for 233 morphological characters for a total of 66 645 observations for 300 taxa. The matrix covers 22 chalcidoid families recognized herein and includes 268 genera within 78 of 83 subfamilies. Morphological data were analysed alone and in combination with molecular data from ribosomal 18S (2105 bp) and 28S D2-D5 expansion regions (1812 bp). Analyses were analysed alone and in combined datasets using implied-weights parsimony and likelihood. Proposed changes in higher classification resulting from the analyses include: (i) recognition of Eriaporidae, revised status; (ii) recognition of Cynipencyrtidae, revised status; (iii) recognition of Azotidae, revised status; (iv) inclusion of Sycophaginae in Agaonidae, revised status; (v) reclassification of Aphelinidae to include Aphelininae, Calesinae, Coccophaginae, Eretmocerinae and Eriaphytinae; (vi) inclusion of Cratominae and Panstenoninae within Pteromalinae (Pteromalidae), new synonymy; (vii) inclusion of Epichrysomallinae in Pteromalidae, revised status. At a higher level, Chalcidoidea was monophyletic, with Mymaridae the sister group of Rotoitidae plus the remaining Chalcidoidea. A eulophid lineage was recovered that included Aphelinidae, Azotidae, Eulophidae, Signiphoridae, Tetracampidae and Trichogrammatidae. Eucharitidae and Perilampidae were monophyletic if Eutrichosomatinae (Pteromalidae) was included, and Eupelmidae was monophyletic if Oodera (Pteromalidae: Cleonyminae) was included. Likelihood recovered a clade of Eupelmidae + (Tanaostigmatidae + (Cynipencyrtus + Encyrtidae). Support for other lineages and their impact on the classification of Chalcidoidea is discussed. Several life-history traits are mapped onto the new phylogeny.

Sexual dimorphism in shells of Cochlostoma septemspirale (Caenogastropoda, Cyclophoroidea, Diplommatinidae, Cochlostomatinae).

Sexual dimorphisms in shell-bearing snails expressed by characteristic traits of their respective shells would offer the possibility for a lot of studies about gender distribution in populations, species, etc. In this study, the seven main shell characters of the snail Cochlostoma septemspirale were measured in both sexes: (1) height and (2) width of the shell, (3) height and (4) width of the aperture, (5) width of the last whorl, (6) rib density on the last whorl, and (7) intensity of the reddish or brown pigments forming three bands over the shell. The variation of size and shape was explored with statistical methods adapted to principal components analysis (PCA) and linear discriminant analysis (LDA). In particular, we applied some multivariate morphometric tools for the analysis of ratios that have been developed only recently, that is, the PCA ratio spectrum, allometry ratio spectrum, and LDA ratio extractor. The overall separation of the two sexes was tested with LDA cross validation.The results show that there is a sexual dimorphism in the size and shape of shells. Females are more slender than males and are characterised by larger size, a slightly reduced aperture height but larger shell height and whorl width. Therefore they have a considerable larger shell volume (about one fifth) in the part above the aperture. Furthermore, the last whorl of females is slightly less strongly pigmented and mean rib density slightly higher. All characters overlap quite considerably between sexes. However, by using cross validation based on the 5 continuous shell characters more than 90% of the shells can be correctly assigned to each sex.

Analysis of ratios in multivariate morphometry.

The analysis of ratios of body measurements is deeply ingrained in the taxonomic literature. Whether for plants or animals, certain ratios are commonly indicated in identification keys, diagnoses, and descriptions. They often provide the only means for separation of cryptic species that mostly lack distinguishing qualitative characters. Additionally, they provide an obvious way to study differences in body proportions, as ratios reflect geometric shape differences. However, when it comes to multivariate analysis of body measurements, for instance, with linear discriminant analysis (LDA) or principal component analysis (PCA), interpretation using body ratios is difficult. Both techniques are commonly applied for separating similar taxa or for exploring the structure of variation, respectively, and require standardized raw or log-transformed variables as input. Here, we develop statistical procedures for the analysis of body ratios in a consistent multivariate statistical framework. In particular, we present algorithms adapted to LDA and PCA that allow the interpretation of numerical results in terms of body proportions. We first introduce a method called the "LDA ratio extractor," which reveals the best ratios for separation of two or more groups with the help of discriminant analysis. We also provide measures for deciding how much of the total differences between individuals or groups of individuals is due to size and how much is due to shape. The second method, a graphical tool called the "PCA ratio spectrum," aims at the interpretation of principal components in terms of body ratios. Based on a similar idea, the "allometry ratio spectrum" is developed which can be used for studying the allometric behavior of ratios. Because size can be defined in different ways, we discuss several concepts of size. Central to this discussion is Jolicoeur's multivariate generalization of the allometry equation, a concept that was derived only with a heuristic argument. Here we present a statistical derivation of the allometric size vector using the method of least squares. The application of the above methods is extensively demonstrated using published data sets from parasitic wasps and rock crabs.