Biological Journal of the Linnean Society

Turtles and tortoises (Order Testudines) possess a unique bony shell that varies in shape across ecological niches. Previous studies have linked turtle shell abnormalities to the presence of environmental stress, leading to asymmetry in shell shape. Here we present the first large-scale geometric morphometric analysis of shell asymmetry in preserved museum specimens from 92 turtle species, using high-resolution 3D scans and (semi)landmark-based methods. We quantified fluctuating asymmetry (FA) and directional asymmetry (DA) in the whole shell, carapace, and plastron, and tested for ecological and phylogenetic influences on shell shape. Our results reveal significant ecological effects on both symmetric and asymmetric components of shell morphology, with aquatic and marine species exhibiting higher FA than their terrestrial counterparts. The carapace showed higher asymmetry and integration than the plastron, suggesting different developmental constraints. Phylogenetic signal was present but weak, indicating convergence in shell shape among ecologically similar but distantly related species. Partial least squares analysis revealed strong covariation between symmetric and asymmetric components, supporting the shell as an integrated morphological unit. These findings highlight the utility of FA as a non-invasive indicator of developmental instability, with implications for conservation monitoring using preserved and living specimens.

The tail of birds contributes substantially to flight aerodynamics through lift generation, reduction of pressure drag, and pitch stabilization. Hummingbirds are powerful flyers, able to sustain hovering in still air, generate lift in both the up- and downstroke, and takeoff due to substantial developmental investment in their wings and corresponding musculature. Given the abundance of wing power, it is possible that tails are less essential to the aerodynamics of hummingbird flight than they are in other birds, freeing them for non-locomotor functions. Hummingbird tails are well known for their morphological elaboration as sexually selected ornaments, including sound generation. Our observations, and brief descriptions in the literature, led us to hypothesize that tail flaring may serve as another form of sexual signaling, used by males during male-male fighting. To test this, we used high-speed video to record agonistic encounters among seven species of hummingbird the field and found 95% of inter- and intra-sexual and specific contests included tail flaring. We measured kinematics of this flaring during male-male fighting of calliope hummingbirds (Selasphorus calliope, n = 5) indoors. Consistent with our hypothesis, captive males exhibited greater angles of tail flare when engaged in a fight (26.9 {+/-} 42.9{degrees}, mean {+/-} sd) than when performing solitary landing (-12.7 {+/-} 8.6 {degrees}) and takeoff (-11.1 {+/-} 6.6 {degrees}) maneuvers. We evaluate these results in the context of signaling during animal contests and propose future tests of whether tail flaring is an honest signal of individual quality and Resource Holding Potential (RHP). Summary StatementMale-male fighting is common in hummingbirds with competition over food and mates. During these competitions, tail flaring and waggle maneuvers are used as a signal of aggressive intent.

Island populations are special for the study of evolutionary processes and can be a zone of incipient speciation. Recently, several island populations of House Wren in the Lesser Antilles (Dominica, St Lucia, St Vincent, and Grenada), formerly recognized as subspecies of the continental form, were reclassified as distinct species. However, much of the supporting data was fragmentary in its sampling of the different islands or equivocal in the patterns observed. Because song is a core element of mate recognition and choice, and can therefore be a key character in species identification, I report here the first detailed characterization and analysis of song for House Wren on all of the islands of the Lesser Antilles where they remain, including Trinidad and Tobago; and compare song patterns across the different islands as well as to several continental populations. Results show that song is broadly similar across all of the islands and to continental populations in high-level features of its structure, organization and delivery but is discriminably different among many of them in its more detailed features. The latter differences are consistent with the recent species splits, with the possible exception of Grenada. They also support retention of House Wren on Trinidad and Tobago as subspecies of the continental form. Results also point to the possibility of a central American origin for some of the islands and a south American origin for others, yielding a trait mosaic where islands that putatively share the same geographic origins, and are therefore presumably genetically closest, are not the most similar in patterns of song (or plumage). This pattern would therefore entail multiple intriguing instances of convergent evolutionary divergence among them that warrants further detailed study. Lay SummaryO_LII provide the first comprehensive analysis and comparison of song patterns of House Wrens for all of the islands of the Lesser Antilles where they remain, some of which are at risk of extirpation, or even extinction if they represent distinct species. C_LIO_LII use the patterns to interpret the recent taxonomic reclassification of many of these island populations as distinct species. C_LIO_LIIn their general structure, organization and delivery male song is similar across all of the islands and follows patterns common to contintental forms of House Wren distributed broadly across North, Central and South America. C_LIO_LISongs of the different islands are, however, discriminably different in their more detailed features and these differences are consistent with most, but possibly not all, of the recent species splits. C_LIO_LIFor the island populations recently reclassified as different species, the distinctiveness of male song is greatest in Dominica and St Vincent and to a lesser extent also St Lucia, and least distinctive in Grenada. Song in Trinidad and Tobago is not substantively different from populations in mainland South America which supports retaining these two island populations as subspecies of the closest continental forms. C_LIO_LISong patterns also point to different possible continental sources for some of the island populations: a source in Central America for Dominica and St Lucia; and a source in South America for the rest. If true, this creates multiple instances of convergent evolutionary divergence in trait patterns across the various islands which merits further study. C_LI

The olfactory system plays a critical role in mammalian environmental perception, with some clades relying on an expanded accessory olfactory (vomeronasal) system (VNS) to detect larger, non-volatile odorants. Mammals make extensive use of this system for social communication between conspecifics. Recent studies have begun to investigate how the VNS changes in response to or as part of ecological transitions. Several studies have identified trends of VNS-associated gene loss or regression in secondarily aquatic mammals. However, continuing discussion on genotype-phenotype correlation within the VNS means that greater effort should be made to investigate the morphology of the VNS in species where it remains poorly understood. Here, we use skeletal and soft-tissue data to demonstrate that the vomeronasal groove, an established osteological correlate for the VNO in bats and primates, is also a valid indicator for its presence in Caniformia. Additionally, we confirm the presence of the VNO in the secondarily aquatic North American river otter (Lontra canadensis) and compare its morphology with that of two close-related species, the semi-aquatic American mink (Neogale vison) and the terrestrial long-tailed weasel (Neogale frenata). This study expands the valid taxonomic scope of the vomeronasal grooves proxy as an osteological correlate, confirms the presence of the VNO in the previously undescribed system of the North American river otter, and highlights the complexity of the mammalian accessory olfactory system.

The evolution of body shape reflects the interplay between functional constraints and habitat structure. In fishes, cave environments are well known for promoting regressive traits such as eye and pigment loss, yet their influence on overall body form remains poorly understood. Here, we examine patterns of body shape variation in cave- and surface-dwelling trichomycterid catfishes from northeastern Colombia to assess whether consistent associations exist between habitat type and morphology. Using geometric morphometric analyses, we quantified differences in body shape among species inhabiting subterranean and surface environments. Our results reveal significant habitat-associated differentiation in body shape along the main axes of morphological variation. Cave-dwelling species exhibit more elongated and fusiform body shapes, whereas surface-dwelling species tend to show deeper and more robust morphologies. In a functional context, these contrasting body patterns suggest associations with differing locomotor demands imposed by subterranean versus surface habitats. Although we do not explicitly test convergence or performance, the recurrence of similar body shapes among species from different clades occupying comparable habitats is consistent with repeated morphological responses to shared ecological constraints. Research HighligthsO_LIMultivariate shape analyses reveal significant habitat-associated variation in trichomycterid fishes. Recurrent morphological patterns suggest repeated responses potentially mediated by habitat constraints. C_LIO_LIBody shape differs consistently between cave- and surface-dwelling trichomycterids. Cave species exhibit more elongated and fusiform forms, whereas surface species display deeper body configurations. C_LI

Islands serve as natural laboratories for exploring evolutionary processes, often fostering unique species through their isolation and distinct ecological conditions. These environments present opportunities to study how a range of selective pressures shape biodiversity. Bird plumage colouration is one trait that has shown to consistently change in island populations, and predation has been hypothesized to influence these differences. While animals often face a trade-off between signalling to conspecifics and avoiding detection by predators, the role of predation in shaping conspicuousness remains underexplored experimentally. In this study, we asked how predation pressure differs between insular and mainland habitats, and whether predation risk covaries with conspicuousness of male and female birds across environments. In a field experiment, we investigated predation rates using 3D-printed models painted to represent both sexes of 12 bird species from three archipelagos (Madeira, Azores, and Canary Islands) and their closest mainland relatives. These models were deployed in the species natural environments to measure hit rates (a proxy for predation risk), accounting for factors that influence prey detectability, such as colour of the models, background contrast, and vegetation. We found that models on the islands experienced less hits compared to those on the mainland, while sexual dichromatic models were more likely to be dislodged on the mainland. In addition, for mainland sites, increased chromatic contrast correlated with a higher probability of dislodgment, suggesting that more conspicuous models were more likely to be hit. These results highlight that while predation constrains conspicuousness, other ecological and evolutionary factors likely drive the reduced plumage colouration observed in island birds. Our research offers experimental insights into how predation interacts with conspicuous traits in shaping plumage colouration in birds.

Sarita Lake, British Columbia houses a distinctive population of threespine stickleback (Gastrosteus aculeatus L.) with a phenotype characterized by unusually large individuals relative to nearby conspecifics. We tested the hypothesis that members of this population are not isometrically larger but rather exhibit variation in allometric trajectories that reflect changes in developmental timing impacting the developmental-genetic architecture of the phenotype. We used 3D geometric morphometrics to characterize the size and shape of skulls, pectoral girdles and pelvic girdles from a sample of individuals from nearby freshwater and marine populations and compare them to a sample from Sarita Lake. We showed that individuals from the Sarita Lake population are larger in each body region compared to most other populations examined. Further, these individuals have dorsally expanded skulls and relatively robust pelvic armour. We also showed that the relationship between size and shape is differently structured among body regions and is heavily influenced by non-uniform sexually-mediated variation across populations sampled. Our results reflect complex underlying developmental trajectories, and we suggest that the large phenotype observed may be driven by fecundity selection on female size in combination with a limnetic trophic niche and relatively increased predation pressure in Sarita Lake.

Arthropods must periodically moult their exoskeleton to permit growth, a conserved developmental process whose morphological and behavioural execution varies widely among lineages. Horseshoe crabs (Limulidae) are members of Xiphosura, a chelicerate lineage with a fossil record extending as far back as the Ordovician and provide a valuable comparative framework for studying the evolution of moulting strategies in Arthropoda. Despite their importance, detailed morphological characterization of moulting in horseshoe crabs remains scarce, limiting developmental studies and broader comparative analyses. Here, we provide a detailed morphological characterization of the moulting process in the Atlantic horseshoe crab Limulus polyphemus. Morphological changes in specific anatomical structures, including the anterior margin of the prosoma, lateral spines and dorsal spinous process of the opisthosoma, were observed during the moulting process. By tracking these morphological markers, such as retraction of the epidermis from the cuticle and degree of corrugation of the epidermis, we were able to identify individuals in the early and late pre-moult stage, predict the onset of ecdysis, and distinguish post-moult and intermoult stages. We compare ecdysis patterns in L. polyphemus with other arthropod taxa, both extant and fossil. We find that, despite differences in behavioural execution, ecdysis in L. polyphemus shares features with other chelicerates, and that both phylogenetic signal and convergent patterns are evident across Arthropoda. This study offers a robust, non-invasive method for determining moult stages in juvenile horseshoe crabs and provides insights into diversity and constraints of ecdysis in Arthropoda.

Sentinel behaviour occurs when individuals use raised positions to scan for predators while the rest of the group forages. Here, we investigated whether a colonial cooperatively breeding species that forages in large groups, the sociable weaver, Philetairus socius, displays sentinel behaviour. This behaviour has been reported in species with similar ecology, behaviour and foraging habits, (e.g. ground foraging in open habitats where aerial predators are common) and, hence, we expected that it could occur in sociable weavers. On the other hand, sentinel behaviour appears to be less common in species that live in very large groups. We used an experimental set-up consisting of an artificial feeding station and perches to assess occurrence of sentinel related behaviours: (i) perching events > 30s on an elevated position, (ii) head-movements and (iii) alarm calling. Birds were seldom observed perching while others fed, and those that did, perched for periods that were too short to be considered as sentinel behaviour (less than 5s on average). Our results suggest that this behaviour is uncommon or even absent in sociable weavers. We discuss whether other factors such as foraging in very large groups, or interspecific foraging associations might make sentinel behaviour less important in this species.

Corvids - members of the crow family - exhibit some of the most sophisticated cognitive abilities outside the primate lineage, yet the developmental origins of many of these abilities remain poorly understood. Here, we present a systematic review of the past 20 years (from 2005) of empirical research on corvid cognitive development, synthesising evidence across core/ foundational, social and physical cognitive domains. Using a structured search strategy and detailed coding framework, we identified 47 relevant studies spanning 16 corvid species. We evaluate and discuss developmental trajectories, species/ taxa-level variation and methodological robustness across studies. For within and between-taxa comparisons, we particularly focus on the best represented abilities in the coded sample: 1) object permanence and caching; 2) tool-use/ manufacture; 3) object manipulation and play; and 4) gaze following. Corvid developmental patterns show both parallels and divergences from those documented in primates and other taxa. However, the existing corvid evidence base is constrained by small samples, captive biases, limited longitudinal data and under-representation of key cognitive abilities, such as executive function, causal reasoning, self-control, metacognition, spatial memory and social learning. We outline critical gaps and future directions, emphasising the need for comparative, longitudinal and ecologically grounded approaches, including the science of magic and Theory of Mind, to better understand how early-life cognition shapes later behaviour, cognition and fitness in this model avian family.

Incilius marmoreus inhabits an extensive range along the Pacific Coast of Mexico and a smaller allopatric region in the State of Veracruz, exhibiting an unusual distribution among herpetofauna. Gunther (1901) classified the Pacific coastal toads Bufo argillaceus and B. lateralis as conspecific with Incilius [Bufo] marmoreus, which has its type locality in Veracruz. Here, we adopt a multidisciplinary approach to reevaluate the phylogeography and taxonomy of I. marmoreus by gathering and analyzing morphological data and conducting phylogenetic and population genetic analyses from genome-wide SNP data. Our results uphold the current taxonomy by concurring with Gunther (1901). Our phylogenetic and population genetic analyses suggest that I. marmoreus from Veracruz are closely related to those from Oaxaca whilst coalescent analyses recovered a north-south split along the Pacific Coast estimated to have occurred [~]0.86 Mya followed by a shallow east-west split in the southern lineage that separates the Pacific coastal populations and the allopatric population in Veracruz [~]0.33 Mya. This species displays marked morphological and genetic diversity throughout its range, but this variation appears to be consistent with gene flow across contiguous populations rather than the existence of independent evolutionary lineages. The processes leading to the geographic isolation of the population on the coast of Veracruz remain uncertain, but we hypothesize that climatic and vegetation changes in the Late Pleistocene may have played a role.

Acoustic signaling is key to individual and species recognition, playing a major role in sexual and social communication. Since reproductive isolation is often maintained through pre-mating mechanisms, song can be an early isolating trait leading to assortative mating, promoting reproductive divergence, and potentially contributing to speciation. However, whether song differences alone are sufficient to prevent interbreeding or if other traits also contribute, remains a matter of debate. Playback experiments provide a more direct way to test the role of song as a reproductive barrier. Here, we use playback experiments to test the hypothesis that song acts as a pre-mating barrier in two recently diverged populations of an island passerine, the Canary Islands Chaffinch (Fringilla canariensis palmae), which inhabit ecologically distinct laurel and pine forests within the island of La Palma. Assuming that male song has diverged in the two habitats, we tested if territorial males from a given habitat responded differently to songs from intruding males from their own habitat or from the other habitat type, using a closely related mainland species as a control. We found that probability of response was weaker to songs of the closely related species and to the different-habitat birds than to songs of the same-habitat birds, but differences for the latter were weak. The intensity of response followed the same pattern. Overall, song divergence between laurel and pine forest chaffinches does not appear strong enough to cause clear behavioural discrimination against individuals from the alternative habitat. Other factors such as morphological and ecological divergence associated with adaptation to local resources might better explain population differentiation. However, testing female responses will be essential to determine whether songs convey lineage-specific information that may elicit assortative mating.

Macromiidae is a widely distributed lineage of libelluloid dragonflies with a largely allopatric genus-level distribution across the Holarctic, Afrotropical, Australasian, and Indo-Malayan regions. Previous studies involving this family have been complicated by morphological convergence and limited phylogenetic sampling. Here, we present the most densely sampled phylogenetic framework for Macromiidae to date, using Anchored Hybrid Enrichment data from 62 of the 125 described species. Our sampling represents all four genera and major geographic regions, including Libelluloid and Cordulegastrid outgroups. Maximum likelihood recovered three major lineages: Epophthalmia, Phyllomacromia, and Macromia sensu lato, with Epophthalmia strongly supported as sister to Phyllomacromia. Didymops was not recovered as monophyletic and was placed within Macromia, although deeper relationships within the Macromia complex showed some gene tree discordance. We additionally scored seven male genitalic characters and reconstructed their evolution across a dated phylogeny. We revealed that these traits varied heavily in phylogenetic signal, with some characters supporting the major clades and others showing high degree of homoplasy. Fossil-calibrated divergence time estimation placed the crown origin of Macromiidae in the late Oligocene (24 Ma), with other major intrafamilial divergences concentrated in the Miocene. Historical biogeographic reconstructions consistently supported Afrotropical origins for Phyllomacromia, Indo-Malayan centered ancestry for Epophthalmia, and a multi-region history for Macromia + Didymops spanning Indo-Malayan, Australasian, and Nearctic regions. Habitat reconstructions favored lentic ancestry for Macromiidae, and diversification rate variation was best explained by trait-independent models rather than lentic/lotic habitat association.

Helping-at-the-nest is often viewed as a precursor to reproduction, but switching between breeder and helper roles has been documented in some species. Such flexibility should depend on the opportunities and benefits of helping, which could differ between sexes due to dispersal strategies and social structure. However, whether breeding-helping flexibility is widespread and sex-specific remains unknown. Here, we investigated sex-specific strategies in breeding-helping flexibility using a 10-year dataset (1955 individuals) on sociable weavers (Philetairus socius), a colonial cooperative breeder in which males are typically philopatric whereas females disperse. Both sexes helped for several years, with males helping more frequently than females and for twice as long (0-13 vs 0-10 years). Around 40% of non-dispersing birds never became breeders and 50% of individuals bred without helping first. Both sexes switched roles between- and within-seasons but males were more likely to alternate (respectively four and six times more than females and up 15 switches across seven years). Our study reveals important flexibility and sex differences in breeder-helper roles, consistent with sex-biased dispersal, spatial breeding proximity and possible indirect fitness benefits. These factors could play a role promoting the evolution of helping across life-stages, not only pre-reproduction, but also reproductive and post-reproductive individuals.

Animal translocations are becoming increasingly popular as a tool for conservationists. Demographic factors can be crucial determinants dictating translocation viability in the short term. Translocated populations pass through artificial bottlenecks and can suffer from founder effects. Reduction in genetic variation relative to their source populations is likely, limiting their adaptive potential. Founder events can increase frequencies of deleterious alleles due to elevated rates of inbreeding and inbreeding depression. Here, we describe the effects of human-driven, serial population translocations on the genetic diversity of critically endangered Anegada iguanas (Cyclura pinguis) in the British Virgin Islands. Though founding populations were extremely small (N=8, N=4), the census sizes of translocated iguana populations increased dramatically over the first twenty years. This implies that these translocations were successful from a demographic perspective despite the small number of animals used, indicating a genetic paradox. To quantify genetic signatures in these bottlenecked populations, blood samples were collected from the source population and two translocated populations and genotyped at 21 microsatellite loci. We found that allele frequencies in translocated populations differed significantly from those of the source, with the translocated populations having less genetic diversity. However, common methods for estimating presence of genetic bottlenecks were non-significant. Estimates of internal relatedness by age class suggest that inbreeding depression may be elevated after translocation, likely reflecting the small initial population sizes associated with these translocation events. Anecdotally, our work shows that translocations may result in subtle genetic erosion that has long-term population viability impacts, even when census size indicates success.

Hypanus brevis (Garman, 1880) and Hypanus dipterurus (Jordan & Gilbert, 1880) are currently considered as a conspecific lineage of the "diamond stingray" from the Eastern Pacific. This taxonomic group has been the subject of nomenclatural disputes for about 145 years. To clarify the historical confusion surrounding this lineage, we employed an integrative taxonomic approach using specimens from the Eastern North and Eastern South Pacific (ENP and ESP). The genetic results, based on single and multilocus mitochondrial analyses, revealed a distinct evolutionary unit in the ESP. While morphological analyses detected subtle differences between ENP and ESP specimens, most characters exhibited significant overlap (e.g., disc shape, dentition patterns, body coloration), suggesting low evolutionary divergence. A calibrated molecular clock analysis estimated this divergence at approximately 3.09 Ma. In accordance with Garmans (1880) original description based on specimens from Paita (northern Peru), we formally resurrect H. brevis from synonymy with H. dipterurus. Our findings suggest an anti-tropical speciation pathway, with core populations of H. brevis and H. dipterurus restricted to the temperate waters of the ESP and ENP, respectively. Notably, a single, fixed COI haplotype was detected in all H. brevis specimens from the north-central Peruvian coast. This result may indicate a severe bottleneck event, raising concerns about the genetic health and long-term viability of this vulnerable species. Finally, we analyzed historical fishery data of H. brevis to infer its current population status, suggesting targeted conservation measures and precautionary management to prevent further loss of genetic diversity.

Historically, venom potencies have been assessed using measures of lethality, such as the median lethal dose (LD50). However, venoms may be selected primarily for their ability to rapidly incapacitate rather than cause mortality, meaning LD50 may not capture the efficacy of venoms in an ecological and evolutionary context. To capture this context, recent studies have adapted measures that assess venoms ability to rapidly incapacitate, such as the median effective dose (ED50). However, while ED50 values are expected to provide a more proximate assessment of ecological variation in venom potency, it is unknown whether historically available LD50 values are still useful proxies of ecologically relevant potency or whether they capture independent axes of venom variation. Here, we test the relationship between LD50 and ED50 in spider venoms by experimentally estimating LD50 and ED50 for 12 species and collating additional potency data for 40 species retrieved from the literature. We observed an isometric relationship between LD50 and ED50 in both analyses, showing these potency measures are both strongly coupled, with an increase in paralysis efficiency associated with a similar increase in lethality. Our results suggest that the functional aspects of venom potency, paralysis and lethality, are intrinsically linked, and due to this strong mechanistic coupling, historically available LD50 values may be used to compare general venom potencies in spiders, provided that they are based on the same prey model.

Island endemic birds are under greater threat than their mainland counterparts. Sedentary living and historically reduced predation may affect island bird sociality and inform their conservation and management. However, detailed studies on their sociality are uncommon. The federally- endangered koloa maoli (Anas wyvilliana, Hawaiian duck, or koloa) is primarily threatened by hybridization with feral mallards and avian botulism outbreaks. We used capture-mark-recapture and genetic data on koloa on the island of Kaua i, a stronghold of remaining koloa, to construct social networks and examine their associations (inferred from co-occurrence in traps) and spatial genetic structure. Information on associations might shed light on preferences for or against mallards and hybrids, and inform planned translocation efforts. Microgeographic spatial genetic structuring where relatedness among individuals scales with geographic distance is a potential liability for maintaining koloa genetic diversity, and would particularly be detrimental during highly localized outbreaks of botulism that could result in the removal of entire lineages. While we found persistent social associations among adult koloa, they were not apparently influenced by plumage traits or body mass, suggesting a lack of social preference for mallard-like individuals. Importantly, we found no spatial patterns of relatedness within the largest refuge. Therefore, botulism outbreaks remain a demographic threat but are unlikely to remove correlated genetic diversity. There were no sex differences in spatial genetic structure and both sexes moved within a refuge. The lack of spatial genetic structure and the presence of many unrelated conspecifics may enable koloa to limit inbreeding and retain genetic diversity without sex-biased dispersal. In the context of future translocations, our results suggest that translocating koloa captured in the same trap together will reduce disruption of preferred associations while also retaining genetic diversity among translocated individuals. LAY SUMMARYO_LIThe koloa maoli (Hawaiian duck, or koloa) is a federally-endangered, island endemic bird. Like other Hawaiian waterbirds, koloa are threatened by introduced predators and habitat loss, but also specifically by hybridization with feral mallards and localized avian botulism outbreaks. Currently, the island of Kaua i has the largest population of non-admixed koloa. We used capture-mark-recapture and genetic data of koloa at two wetland sites on Kaua i to examine their sociality and spatial genetic structure. C_LIO_LIKoloa formed preferential social associations, but they were not based on plumage traits, body mass or genetic relatedness. C_LIO_LIThere was no spatial genetic structure for males and females within a wetland site. Our results suggested that 1) koloa have no preference for mallard-like plumage that might increase hybrid pairing, 2) localized (within-refuge) botulism outbreaks are unlikely to remove close relatives and unique genetic lineages, and 3) translocation of groups could maintain social associations without limiting genetic diversity. C_LI

Understanding morphological variation is crucial for the study of speciation and for conservation as it helps in assessing biodiversity and predicting responses to environmental changes. These approaches are broadly applicable but are especially valuable in marine environments, where species are often elusive, difficult to study, and face heightened threats from rapid environmental shifts. The marine snail Littorina saxatilis is notable for its extensive polymorphism in shell shape, size, and colour, with ecotypes that evolve in response to environmental forces including wave exposure and crab predation. Morphometric tools have been central to investigating the mechanisms driving this phenotypic divergence; yet, a direct comparison of their methodological efficacy is lacking. Here, we took advantage of L. saxatilis ecotypes to contrast three morphometric approaches: elliptical Fourier analysis (EFA), landmarks-based geometric morphometrics (GM), and the growth-based model implemented in the ShellShaper software (SS). We assessed their clustering power, biological interpretability, robustness to measurement error and transferability among datasets. Our findings provide insights to guide method selection in studies aimed at exploring morphological variation: EFA is better suited for high-throughput screening and describing intermediate shapes; SS offers superior clustering power with highly interpretable growth parameters; and GM is best for detailed anatomical studies but is less efficient for large datasets. We provide guidelines to align method selection with specific research goals, balancing analytical efficiency with the required morphological and biological insight. By following this framework, researchers can ensure that robust morphological analysis is achieved, which is essential not only for elucidating mechanisms of adaptation and speciation but also for effective management and conservation of marine biodiversity.

The connection between female host plant preference and offspring performance is important for understanding how relationships between plants and phytophagous insects have evolved. According to the preference-performance hypothesis, female insects should evolve to oviposit on host plants on which offspring performance is the highest. Here, we examined the preference-performance hypothesis in the northern brown argus (Aricia artaxerxes) butterfly in the province of Uppland, Sweden, by comparing female host plant preference and larval growth between the host plant species wood cranesbill (Geranium sylvaticum) and bloody cranesbill (G. sanguineum). We also investigated if host plant preference in A. artaxerxes was related to the geographic distribution of A. artaxerxes and its host plants in the province Uppland. We found that the A. artaxerxes females, contrary to the preference-performance hypothesis, preferred ovipositing on G. sylvaticum, even though larvae feeding on G. sylvaticum were slightly smaller than those feeding on G. sanguineum. Since G. sylvaticum is more abundant and probably more utilized than G. sanguineum in Uppland, an explanation for this negative preference-performance connection may be that there are advantages associated with utilizing a more common host plant species, even though larvae feeding on this plant show reduced growth rates. Overall, the results show that factors other than offspring performance, such as geographic distribution, may influence female host plant preference in A. artaxerxes.