A Comparative Study of Body Size Evolution in Moths: Evidence of Correlated Evolution with Feeding and Phenology-Related Traits.

Interspecific variation in body size is one of the most popular topics in comparative studies. Despite recent advances, little is still known about the patterns and processes behind the evolution of body size in insects. Here, we used a robust data set comprising all geometrid moth species occurring in Northern Europe to examine the evolutionary associations involving body size and several life-history traits under an explicitly phylogenetic framework. We provided new insights into the interactive effects of life-history traits on body size and evidence of correlated evolution. We further established the sequence of trait evolution linking body size with the life-history traits correlated with it. We found that most (but not all) of the studied life-history traits, to some extent, interfered with interspecific variation in body size, but interactive effects were uncommon. Both bi- and multivariate phylogenetic analyses indicated that larger species tend to be nocturnal flyers, overwinter in the larval stage, feed on the foliage of trees rather than herbs, and have a generalist feeding behavior. We found evidence of correlated evolution involving body size with overwintering stage, host-plant growth form, and dietary specialization. The examination of evolutionary transitions within the correlated models signaled that overwintering as larvae preceded the evolution of large sizes, as did feeding on tree foliage and the generalist feeding behavior. By showing that both body size and all life-history traits correlated with it evolve at very slow rates, we caution against uncritical attempts to propose causal explanations for respective associations based on contemporary ecological settings.

Evolution of wing shape in geometrid moths: phylogenetic effects dominate over ecology.

Locomotory performance is an important determinant of fitness in most animals, including flying insects. Strong selective pressures on wing morphology are therefore expected. Previous studies on wing shape in Lepidoptera have found some support for hypotheses relating wing shape to environment-specific selective pressures on aerodynamic performance. Here, we present a phylogenetic comparative study on wing shape in the lepidopteran family Geometridae, covering 374 species of the northern European fauna. We focused on 11 wing traits including aspect ratio, wing roundness, and the pointedness of the apex, as well as the ratio of forewing and hindwing areas. All measures were taken from images available on the internet, using a combination of tools available in Fiji software and R. We found that wing shape demonstrates a phylogenetically conservative pattern of evolution in Geometridae, showing similar or stronger phylogenetic signal than many of its potential predictors. Several wing traits showed statistically significant associations with predictors such as body size, phenology, and preference for forest habitats. Overall, however, all of these associations remained notably weak, with no wing shape being excluded for any value of the predictors, including body size. We conclude that, in geometrids, wing traits do not readily respond to selective pressures optimizing aerodynamic performance of the moths in different environments. Selection on wing shape may nevertheless operate through other functions of the wings, with the effectiveness of crypsis at rest being a promising candidate for further studies.

Molecular phylogeny, classification, biogeography and diversification patterns of a diverse group of moths (Geometridae: Boarmiini).

Understanding how and why some groups have become more species-rich than others, and how past biogeography may have shaped their current distribution, are questions that evolutionary biologists have long attempted to answer. We investigated diversification patterns and historical biogeography of a hyperdiverse lineage of Lepidoptera, the geometrid moths, by studying its most species-rich tribe Boarmiini, which comprises ca. 200 genera and ca. known 3000 species. We inferred the evolutionary relationships of Boarmiini based on a dataset of 346 taxa, with up to eight genetic markers under a maximum likelihood approach. The monophyly of Boarmiini is strongly supported. However, the phylogenetic position of many taxa does not agree with current taxonomy, although the monophyly of most major genera within the tribe is supported after minor adjustments. Three genera are synonymized, one new combination is proposed, and four species are placed in incertae sedis within Boarmiini. Our results support the idea of a rapid initial diversification of Boarmiini, which also implies that no major taxonomic subdivisions of the group can currently be proposed. A time-calibrated tree and biogeographical analyses suggest that boarmiines appeared in Laurasia ca. 52 Mya, followed by dispersal events throughout the Australasian, African and Neotropical regions. Most of the transcontinental dispersal events occurred in the Eocene, a period of intense geological activity and rapid climate change. Diversification analyses showed a relatively constant diversification rate for all Boarmiini, except in one clade containing the species-rich genus Cleora. The present work represents a substantial contribution towards understanding the evolutionary origin of Boarmiini moths. Our results, inevitably biased by taxon sampling, highlight the difficulties with working on species-rich groups that have not received much attention outside of Europe. Specifically, poor knowledge of the natural history of geometrids (particularly in tropical clades) limits our ability to identify key innovations underlying the diversification of boarmiines.

Quantifying the effects of species traits on predation risk in nature: A comparative study of butterfly wing damage.

Evading predators is a fundamental aspect of the ecology and evolution of all prey animals. In studying the influence of prey traits on predation risk, previous researchers have shown that crypsis reduces attack rates on resting prey, predation risk increases with increased prey activity, and rapid locomotion reduces attack rates and increases chances of surviving predator attacks. However, evidence for these conclusions is nearly always based on observations of selected species under artificial conditions. In nature, it remains unclear how defensive traits such as crypsis, activity levels and speed influence realized predation risk across species in a community. Whereas direct observations of predator-prey interactions in nature are rare, insight can be gained by quantifying bodily damage caused by failed predator attacks. We quantified how butterfly species traits affect predation risk in nature by determining how defensive traits correlate with wing damage caused by failed predation attempts, thereby providing the first robust multi-species comparative analysis of predator-induced bodily damage in wild animals. For 34 species of fruit-feeding butterflies in an African forest, we recorded wing damage and quantified crypsis, activity levels and flight speed. We then tested for correlations between damage parameters and species traits using comparative methods that account for measurement error. We detected considerable differences in the extent, location and symmetry of wing surface loss among species, with smaller differences between sexes. We found that males (but not females) of species that flew faster had substantially less wing surface loss. However, we found no correlation between cryptic coloration and symmetrical wing surface loss across species. In species in which males appeared to be more active than females, males had a lower proportion of symmetrical wing surface loss than females. Our results provide evidence that activity greatly influences the probability of attacks and that flying rapidly is effective for escaping pursuing predators in the wild, but we did not find evidence that cryptic species are less likely to be attacked while at rest.

No Indication of High Host-Plant Specificity in Afrotropical Geometrid Moths.

Specificity is one of the fundamental concepts in ecology. Host specificity of phytophagous insects has been of particular interest because of its crucial role in diversification and life-history evolution. However, the majority of tropical insects remain insufficiently explored with respect to their host-plant relations. A lack of respective data is also hindering the debate over whether higher levels of host-plant specificity prevail in tropical insects compared to temperate ones. We investigated host-plant specificity of forest geometrid moths (Lepidoptera: Geometridae) in equatorial Africa using host-plant acceptability trials with neonate larvae, with the addition of field observations. We compare our experimental data to the (well-known) host-specificity patterns of closely related temperate (hemiboreal) species. Similarly to the temperate region, there were broadly polyphagous tropical species in several clades of Geometridae utilizing hosts belonging to different plant families. Phylogenetic comparative analysis returned no significant differences in host specificity between the two regions. Our study contributes to the evidence that host-plant specificity of herbivores is not necessarily substantially higher in tropical than temperate regions.

Day-flying moths are smaller: evidence for ecological costs of being large.

Research on evolutionary forces determining optimal body sizes has primarily relied on experimental evaluation of respective selective pressures. Accounting for among-species variation through application of phylogenetic comparative methods is a complementary although little used approach. It enables the direct association of body size values with particular environments. Using phylogenetically explicit comparative analyses, we show that small body size is associated with diurnal (rather than nocturnal) activity of adults among temperate species of the moth family Geometridae. The association of an exclusively adult trait with species-specific body size suggests that optimal body sizes are at least partly determined by the costs being a large adult, as opposed to the more frequently considered costs of attaining large size. It appears likely that size-selective predation by insectivorous birds is the primary factor responsible for selection against large body size in day-flying moths.

An ordination of life histories using morphological proxies: capital vs. income breeding in insects.

Predictive classifications of life histories are essential for evolutionary ecology. While attempts to apply a single approach to all organisms may be overambitious, recent advances suggest that more narrow ordination schemes can be useful. However, these schemes mostly lack easily observable proxies of the position of a species on respective axes. It has been proposed that, in insects, the degree of capital (vs. income) breeding, reflecting the importance of adult feeding for reproduction, correlates with various ecological traits at the level of among-species comparison. We sought to prove these ideas via rigorous phylogenetic comparative analyses. We used experimentally derived life-history data for 57 species of European Geometridae (Lepidoptera), and an original phylogenetic reconstruction. The degree of capital breeding was estimated based on morphological proxies, including relative abdomen size of females. Applying Brownian-motion-based comparative analyses (with an original update to include error estimates), we demonstrated the associations between the degree of capital breeding and larval diet breadth, sexual size dimorphism, and reproductive season. Ornstein-Uhlenbeck model based phylogenetic analysis suggested a causal relationship between the degree of capital breeding and diet breadth. Our study indicates that the gradation from capital to income breeding is an informative axis to ordinate life-history strategies in flying insects which are affected by the fecundity vs. mobility trade off, with the availability of easy to record proxies contributing to its predictive power in practical contexts.

Morphological and molecular data reveal fifteen new sympatric Zygomyia Winnertz, 1864 (Diptera: Mycetophilidae) species from Ecuador.

Fifteen new Zygomyia Winnertz species are described from material collected from one location in Ecuador during a very short time period: Z. acrotricha sp. nov., Z. comata sp. nov., Z. diversiseta sp. nov., Z. flamma sp. nov., Z. geniculata sp. nov., Z. longistyla sp. nov., Z. napoensis sp. nov., Z. paniculata sp. nov., Z. pectinella sp. nov., Z. spinosa sp. nov., Z. talpaeforma sp. nov., Z. trilobata sp. nov., Z. triseta sp. nov., Z. uniordata sp. nov. and Z. ximoi sp. nov. Zygomyia bicolor Edwards, 1934 is redescribed, including detailed illustration of the male terminalia. The number of Neotropical Zygomyia species is increased to 30. An updated key to the Neotropical Zygomyia is provided. Unexpectedly high level of interspecific genetic variation between Neotropical Zygomyia spp. is reported.

A supertree of Northern European macromoths.

Ecological and life-history data on the Northern European macromoth (Lepidoptera: Macroheterocera) fauna is widely available and ideal for use in answering phylogeny-based research questions: for example, in comparative biology. However, phylogenetic information for such studies lags behind. Here, as a synthesis of all currently available phylogenetic information on the group, we produce a supertree of 114 Northern European macromoth genera (in four superfamilies, with Geometroidea considered separately), providing the most complete phylogenetic picture of this fauna available to date. In doing so, we assess those parts of the phylogeny that are well resolved and those that are uncertain. Furthermore, we identify those genera for which phylogenetic information is currently too poor to include in such a supertree, or entirely absent, as targets for future work. As an aid to studies involving these genera, we provide information on their likely positions within the macromoth tree. With phylogenies playing an ever more important role in the field, this supertree should be useful in informing future ecological and evolutionary studies.

The northernmost record of a blood-sucking ectoparasite, Lipoptena fortisetosa Maa (Diptera: Hippoboscidae), in Estonia.

BACKGROUND: Deer keds are obligatory haematophagous parasites of large homeothermic animals, particularly cervids. Two of the five known species occurring in Europe-Lipoptena cervi (Linnaeus) and L. fortisetosa Maa-are known to have a relatively wide distribution. Lipoptena fortisetosa is considered to have been introduced into Europe with sika deer from the Eastern Palaearctic and is continuously expanding its range. Little is known about the medical importance of deer keds, but they can cause hair loss in cervids and are suspected to be vectors of several diseases. NEW INFORMATION: Details of the distribution of Lipoptena fortisetosa in Europe, including its northernmost record, are provided. This species has been shown to have a viable population in Southern Estonia. Furthermore, the differences from allied L. cervi are discussed, based on morphological and molecular characters.

A comprehensive molecular phylogeny of Geometridae (Lepidoptera) with a focus on enigmatic small subfamilies.

Our study aims to investigate the relationships of the major lineages within the moth family Geometridae, with a focus on the poorly studied Oenochrominae-Desmobathrinae complex, and to translate some of the results into a coherent subfamilial and tribal level classification for the family. We analyzed a molecular dataset of 1,206 Geometroidea terminal taxa from all biogeographical regions comprising up to 11 molecular markers that includes one mitochondrial (COI) and 10 protein-coding nuclear gene regions (wingless, ArgK, MDH, RpS5, GAPDH, IDH, Ca-ATPase, Nex9, EF-1alpha, CAD). The molecular data set was analyzed using maximum likelihood as implemented in IQ-TREE and RAxML. We found high support for the subfamilies Larentiinae, Geometrinae and Ennominae in their traditional scopes. Sterrhinae becomes monophyletic only if Ergavia Walker, Ametris Hübner and Macrotes Westwood, which are currently placed in Oenochrominae, are formally transferred to Sterrhinae. Desmobathrinae and Oenochrominae are found to be polyphyletic. The concepts of Oenochrominae and Desmobathrinae required major revision and, after appropriate rearrangements, these groups also form monophyletic subfamily-level entities. Oenochrominae s.str. as originally conceived by Guenée is phylogenetically distant from Epidesmia and its close relatives. The latter is hereby described as the subfamily Epidesmiinae Murillo-Ramos, Sihvonen & Brehm, subfam. nov. Epidesmiinae are a lineage of "slender-bodied Oenochrominae" that include the genera Ecphyas Turner, Systatica Turner, Adeixis Warren, Dichromodes Guenée, Phrixocomes Turner, Abraxaphantes Warren, Epidesmia Duncan & Westwood and Phrataria Walker. Archiearinae are monophyletic when Dirce and Acalyphes are formally transferred to Ennominae. We also found that many tribes were para- or polyphyletic and therefore propose tens of taxonomic changes at the tribe and subfamily levels. Archaeobalbini stat. rev. Viidalepp (Geometrinae) is raised from synonymy with Pseudoterpnini Warren to tribal rank. Chlorodontoperini Murillo-Ramos, Sihvonen & Brehm, trib. nov. and Drepanogynini Murillo-Ramos, Sihvonen & Brehm, trib. nov. are described as new tribes in Geometrinae and Ennominae, respectively.

Sex Chromosomes of the Iconic Moth Abraxas grossulariata (Lepidoptera, Geometridae) and Its Congener A. sylvata.

The magpie moth, Abraxas grossulariata, is an iconic species in which female heterogamety was discovered at the beginning of the 20th century. However, the sex chromosomes of this species have not yet been cytologically identified. We describe the sex chromosomes of A. grossulariata and its congener, A. sylvata. Although these species split only around 9.5 million years ago, and both species have the expected WZ/ZZ chromosomal system of sex determination and their sex chromosomes share the major ribosomal DNA (rDNA) representing the nucleolar organizer region (NOR), we found major differences between their karyotypes, including between their sex chromosomes. The species differ in chromosome number, which is 2n = 56 in A. grossularita and 2n = 58 in A. sylvata. In addition, A. grossularita autosomes exhibit massive autosomal blocks of heterochromatin, which is a very rare phenomenon in Lepidoptera, whereas the autosomes of A. sylvata are completely devoid of distinct heterochromatin. Their W chromosomes differ greatly. Although they are largely composed of female-specific DNA sequences, as shown by comparative genomic hybridization, cross-species W-chromosome painting revealed considerable sequence differences between them. The results suggest a relatively rapid molecular divergence of Abraxas W chromosomes by the independent spreading of female-specific repetitive sequences.

Epirrita pulchraria (Taylor, 1907) transferred to Malacodea, with notes on the phylogeny and ecology of the tribe Operophterini (Lepidoptera: Geometridae: Larentiinae).

The Nearctic Epirrita pulchraria (Taylor, 1907) was revealed as a sister taxon of the Palaearctic Malacodea regelaria Tengström,1869 in a molecular phylogenetic analysis of the tribe Operophterini. DNA sequence variation in one mitochondrial (COI) and eight nuclear (28S, EF-1α, WGL, GAPDH, RPS5, IDH, MDH, CAD) gene fragments was used in the analysis. Bayesian inference resulted in a well-resolved phylogenetic hypothesis. According to the recovered phylogeny a new combination is proposed: Rachela pulchraria Taylor, 1907 is transferred from Epirrita Hübner, 1808 to Malacodea Tengström, 1869 as Malacodea pulchraria (Taylor, 1907) comb. nov. Larvae of both M. pulchraria and M. regelaria feed on coniferous trees, which distinguishes them from other members of the tribe. Close relatedness of M. pulchraria and M. regelaria is also supported by our study of male genital morphology. Ambiguous phylogenetic affinities of Epirrita viridipurpurescens (Prout, 1937) are discussed.

Two new Neuratelia Rondani (Diptera, Mycetophilidae) species from Western Palaearctic: a case of limited congruence between morphology and DNA sequence data.

Two new Mycetophilidae species, Neurateliajabalmoussae sp. n. and Neurateliasalmelai sp. n. are described on the basis of material collected from Lebanon, Estonia and Finland. Detailed figures of male terminalia and photographs of general facies are provided along with discussions of their morphological distinction from sibling species. For the first time molecular characters are used to distinguish new fungus gnat species. Molecular analysis relies on cytochrome oxidase subunit one (COI) but has additionally been corroborated by information from the 28S and ITS2 regions of nuclear ribosomal DNA. Situations where morphological and molecular data provide conflicting evidence for species delimitation are discussed. A new country record from Georgia is provided for Neurateliacaucasica.

Degree of specialization is related to body size in herbivorous insects: a phylogenetic confirmation.

Numerous studies have suggested a general relationship between the degree of host specialization and body size in herbivorous animals. In insects, smaller species are usually shown to be more specialized than larger-bodied ones. Various hypotheses have attempted to explain this pattern but rigorous proof of the body size-diet breadth relationship has been lacking, primarily because the scarceness of reliable phylogenetic information has precluded formal comparative analyses. Explicitly using phylogenetic information for a group of herbivores (geometrid moths) and their host plant range, we perform a comparative analysis to study the body size-diet breadth relationship. Considering several alternative measures of body size and diet breadth, our results convincingly demonstrate without previous methodological issues-a first for any taxon-a positive association between these traits, which has implications for evaluating various central aspects of the evolutionary ecology of herbivorous insects. We additionally demonstrate how the methods used in this study can be applied in assessing hypotheses to explain the body size-diet breadth relationship. By analyzing the relationship in tree-feeders alone and finding that the positive relationship remains, the result suggests that the body size-diet breadth relationship is not solely driven by the type of host plant that species feed on.

Baeopterogyna mihalyii Matile (Diptera, Mycetophilidae): association of sexes using morphological and molecular approaches with the first description of females.

Both males and females of Baeopterogyna mihalyiiMatile, 1975 are recorded from northern Greece. Females are described for the first time providing photographs of the general facies and terminalia. In contrast to the single congener with stenopterous females - Baeopterogyna nudipes Vockeroth, 1972 - Baeopterogyna mihalyii is shown to have normally developed wings in both sexes. Association of sexes is based on both morphological characters and sequence data from cytochrome oxidase subunit one (COI). DNA sequences are used for the first time for the association of sexes in Mycetophilidae.