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Zoological Journal of the Linnean Society

◐ Oxford University Press (OUP)

Preprints posted in the last 30 days, ranked by how well they match Zoological Journal of the Linnean Society's content profile, based on 18 papers previously published here. The average preprint has a 0.02% match score for this journal, so anything above that is already an above-average fit.

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First confirmed record in Melilla of Gerbillus campestris Loche, 1867 (Muridae: Gerbillinae) for Spain

Ferrandez Verdu, T.; Ruiz, F. P.; Carrillo Lopez, A. F.; Tapia Caro, M.; Castillo Gonzalez, A.; Vives Lopez, R.; Amoros Box, C.; Marcos Perez, M.; Boratynski, Z.; Pavlickova, B.

2026-10-06 zoology 10.64898/2026.09.30.754369 medRxiv
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The fieldwork was carried out between December 2023 and June 2025, with the aim, among others, of confirming the possible presence of the North African Gerbil (Gerbillus campestris) in Melilla. To this end, 56 camera trap stations were installed, each equipped with a camera trap. Of the total number of camera trap stations installed, the presence of Gerbillus campestris was detected in six of them, representing a capture success rate of 10.71%. Following the detection of the species, a specimen was captured using cage traps. For identification purposes, samples of epithelial tissue from the ear and faeces were collected. These samples were sent to the Institute of Vertebrate Biology of the Czech Academy of Sciences, Brno, Czech Republic, for analysis. There, DNA was extracted from the samples using the Innu PREP DNA Mini Kit 2.0 (IST Innuscreen GmbH, Germany) according to the manufacturer's protocol. The results obtained confirm, for the first time through records of the species, the presence of Gerbillus campestris in Melilla and, therefore, in Spain and Europe.

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The allometry of spiracles and gill slits: a window into the respiratory capacity of rays

Aitchison, R. M.; VanderWright, W. J.; Arnold, A. E.; Iliou, A. S.; Neufeld, B.; Vuorensivu, L.; Bryan, S. R.; Kui, O.; Matsushiba, J. H.; Wynn Dulvy, O.; Wynn Dulvy, S.; Dulvy, N. K.

2026-09-30 ecology 10.64898/2026.09.29.755487 medRxiv
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Metabolic rate governs the pace of life through the allocation and expenditure of energy, yet metabolic rate data are unavailable for most species. External morphological traits, such as gill slits, have been used to infer respiratory physiology and metabolism in sharks, yet, whether that pattern is generalizable across rays is unknown. In rays (Batoidea), spiracle length seems adaptive to oxygen uptake capacity, environmental oxygen availability, and activity level. Thus, we evaluate the allometry of external respiratory morphology (spiracles and gill slits) in rays using 550 museum specimens representing 88 species. We test whether spiracle length and gill slit height grow isometrically with body size (total length or disc width) within and across species. Overall, we find spiracles and gill slits are morphologically diverse, varying widely in shape within and between families. Spiracle length and gill slit height scale isometrically and are a consistent proportion of body size, despite such high morphological variability. While that proportion is consistent through ontogeny, each species proportion differs (e.g. gill slit height ranges from 0.93% of total length in Spotted Legskate to 2.93% in Pacific Electric Ray). Of the three swimming styles, pectoral oscillating species (Mobulid and Eagle Rays) may have steeper scaling of spiracle length with gill slit height - consistent with greater respiratory capacity in species with more active lifestyles. Our findings are consistent with literature hypothesizing external respiratory morphology (spiracles and gill slits) is tightly related to metabolic rate and activity level, and demonstrate the value of natural history collections in ecological research. Importantly, the isometry of both respiratory traits means that a single measure of any individual, irrespective of size, will be representative of the species thus providing a basis for expanding our understanding of metabolic morphology, enabling future work on life history, extinction risk, and ecological function.

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Insights into the molecular systematics of montane voles of the Western Himalayan Mountains

Khan, M. R.; Nadeem, M. S.; Coleman, J.; Cannatella, D.

2026-10-07 evolutionary biology 10.64898/2026.10.02.756046 medRxiv
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Kashmir voles (Hyperacrius) are poorly studied fossorial arvicolines of the southern Himalayan slopes, with two species recognized. We sequenced Cytb from voles collected in northern Pakistan and combined these data with existing GenBank sequences, yielding 194 tips spanning nearly all arvicoline genera. We analyzed five data sets with maximum likelihood and compared external measurements among species using principal component and linear discriminant analyses. We recovered five well-supported clades of Hyperacrius separated by ~12-14% p-distance, which we treat as distinct species. We recognize clade 1 as H. wynnei, and populations referred to H. w. traubi are genetically indistinguishable from H. w. wynnei. We recognize H. zygomaticus, new rank (clade 2), and tentatively allocate clade 5 to H. brachelix. Clades 3 and 4 likely represent undescribed species. Morphometric analyses of external measurements clearly distinguish H. wynnei but not clades 3 and 4 (overall classification accuracy 70.3%). Phylogenetic analyses including nuclear genes placed Hyperacrius as sister to a clade containing Microtini, Arvicola, Lagurini, Dinaromys, and Ellobiusini, whereas mitochondrial analyses weakly allied it with Ellobiusini. Its traditional placement in Clethrionomyini is thus unsupported, although its exact position remains unresolved. Alticola is nonmonophyletic, with A. macrotis sequences nested within Clethrionomys, likely reflecting mitochondrial capture.

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Pseudocryptic diversity within the Phakellia robusta species complex: description of Phakellia pallida sp. nov. and Phakellia pusilla sp. nov.

Gracia-Sancha, C.; Taboada, S.; De la Torriente, A.; Galia-Camps, C.; Gonzalez-Porto, M.; Rios, P.; Cristobo, J.; Serrano, A.; Massuti, E.; Riesgo, A.; Diaz, J. A.

2026-10-07 zoology 10.64898/2026.10.06.756958 medRxiv
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The genus Phakellia comprises predominantly fan-shaped deep-sea sponges, but its species diversity and boundaries remain unresolved. Here, we use an integrative approach combining morphological and molecular data to investigate the Phakellia robusta species complex and describe two new species, Phakellia pallida sp. nov. and Phakellia pusilla sp. nov. Overlap in external and spicular morphology makes these taxa challenging to identify. Molecular analyses based on mitochondrial (cox1) and nuclear (28S) markers support the recognition of the three taxa as distinct species, and reveal their close evolutionary relationship. The two new species co-occur in the NE Atlantic Ocean and the Mediterranean Sea, with overlapping depth ranges, although P. pallida sp. nov. is generally found at greater depths. Our results also suggest that several previous records of P. robusta from the Mediterranean and southern parts of the NE Atlantic may instead correspond to either P. pallida sp. nov. or P. pusilla sp. nov., indicating that its distribution requires reassessment. In addition, a group I intron previously reported in the cox1 gene of P. robusta was polymorphically detected at the same insertion site in some, but not all, individuals of P. pusilla sp. nov., but it was absent from P. pallida sp. nov. individuals. Overall, our results reveal pseudocryptic diversity within Phakellia and highlight the value of integrative approaches for resolving species boundaries and reassessing deep-sea sponge diversity and distributions.

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Decoupled wing evolution reveals trait-specific links between morphology, geography, and diversification in Ithomiini butterflies.

Chazot, N.; Dore, M.; Mathou, A.; Willmott, K. R.; Lagarde, S.; Freitas, A. V. L.; Cornette, R.; Elias, M.

2026-09-28 ecology 10.64898/2026.09.25.754348 medRxiv
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Understanding when species diversification and phenotypic evolution are coupled remains a central challenge in macroevolution. Such coupling may depend on the traits considered, phenotypic integration, or the geographic context in which diversification occurs. Here, we investigate these relationships in Ithomiini, a diverse Neotropical butterfly tribe whose diversification is strongly associated with the Andes-Amazonia system. We quantified forewing and hindwing size and shape for 1,618 individuals representing 291 species using geometric morphometrics, and combined these data with phylogenetic comparative methods, diversification-rate estimates, and spatial analyses of species distributions. Evolutionary rate shifts were largely trait- and wing-specific, indicating limited coupling between forewing and hindwing evolution. Across the phylogeny, wing size evolution and extant wing size were not associated with species diversification, nor was hindwing shape evolution. By contrast, forewing shape evolutionary rates showed a positive association with diversification. Spatial analyses revealed that this coupling was geographically concentrated in the Andes, where high diversification rates coincided with elevated forewing size and shape diversity and increased forewing evolutionary rates. Hindwing diversity, by contrast, more closely tracked phylogenetic diversity and showed weaker or opposing relationships with diversification. Our results show that diversification and phenotypic evolution in Ithomiini are neither generally coupled nor fully independent, but linked in trait-specific and geographically structured ways. More broadly, they highlight the need to decompose complex phenotypes and integrate spatial context when testing macroevolutionary coupling.

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Bill colour outperforms iris colour for sexing in Starlings Sturnus vulgaris in breeding condition

Pascual, K.; Bateson, M.

2026-09-30 zoology 10.64898/2026.09.28.755006 medRxiv
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Reliable morphological criteria for sexing birds without the need for invasive procedures are scientifically and ethically valuable. Smith et al (2005) reported that iris colour assigns the genetic sex of Starlings Sturnus vulgaris with 98% accuracy, and bill base colour with 100% accuracy when birds are in breeding condition, but these figures have not been tested independently. We applied their criteria to 31 captive Starlings in breeding condition, using three observers blind to genetic sex, two of them new to Starling sexing. Bill base colour correctly predicted genetic sex for all 29 birds scored, with complete observer agreement. Mean iris score was correct for 27 of 31 (87.1%), fewer than the 94 of 96 reported by Smith et al (2005) for birds in breeding condition (Fisher exact test, P = 0.031), although with so few errors in either study this comparison is imprecise. Observer agreement was high, so these errors reflect iris colouration conflicting with genetic sex rather than inconsistent scoring; bill base colour was correct for all four misassigned birds. Body mass and tarsus length classified 77% and 58% of birds correctly, their published thresholds missing in opposite directions. We conclude that bill base colour should be the primary criterion for sexing Starlings in breeding condition; the accuracy of iris colour should not be assumed to be 98%.

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Morphological evolution and phylogeny are unlinked in Hawaiian Doryopteris (Cheilanthoideae: Pteridaceae)

Lau, P.; Zhang, X.; Landis, J. B.; Burleigh, J. G.; Huiet, L.; Wood, K. R.; Thomas, M. K.; Zenil-Ferguson, R.; Rothfels, C. J.; Tribble, C. M.

2026-10-09 evolutionary biology 10.64898/2026.10.06.757157 medRxiv
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Premise of the study: Doryopteris J.Sm. is a genus of dry-adapted ferns found widely across the southern hemisphere and the tropics. In the Hawaiian Islands, the genus is represented by four endemic species and one putative hybrid. Two of the endemic species (D. decora and D. decipiens) are widespread across all major islands, while two (D. angelica and D. takeuchii) are federally and state-listed endangered single-island endemics.We aimed to resolve the evolutionary relationships and biogeographic history of Hawaiian Doryopteris. Methods: We used a target-capture approach and the GoFlag 408 flagellate land plant probe set to sequence more than 400 nuclear loci for 40 Hawaiian and 29 non-Hawaiian individuals in Doryopteris and the related genera Lytoneuron and Ormopteris. After processing the sequences into alignments and single nucleotide polymorphism sites, we performed phylogenetic inference and population-level principal component analyses. Key results: We infer that Hawaiian Doryopteris are monophyletic and are nested within the pantropical species D. concolor. Within Hawai'i, clades and clusters of individuals correspond to geographical, geological, and climatological factors, more so than species identifications, with a few exceptions. We posit that Hawaiian Doryopteris represents an incipient radiation at the very earliest stages of divergence, and that budding speciation has led to non-monophyly of several taxa. Conclusions Our inference using genome-wide sequencing data sheds light on the complex evolutionary history of Hawaiian Doryopteris. Overall, the phylogeny of Hawaiian Doryopteris shows strong links with geographical, geological, and climatological processes and little with morphological evolution.

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Evolutionary rates in "living fossils" across the Tree of Life

Cascini, M.; Lopez Estrada, E. K.; Duchene, D. A.; Ho, S.

2026-10-07 evolutionary biology 10.64898/2026.10.03.756400 medRxiv
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Lineages that have undergone little apparent evolutionary change over long geological intervals are often described as "living fossils", with iconic examples including the coelacanth, tuatara, and Ginkgo. However, studies of morphological and molecular data have produced mixed evidence regarding whether living fossils have evolved more slowly than their relatives. We tested for reduced evolutionary rates using a sister-pairs approach, analysing a comprehensive set of morphological and molecular data for 45 living-fossil lineages and their close relatives across the Tree of Life. Living-fossil lineages showed substantially reduced rates of morphological evolution, whereas evidence for reduced molecular rates was weaker. To test whether this pattern could be explained by species poverty, we also analysed other depauperate lineages (species-poor clades contrasted with species-rich sister clades) and found a weak tendency towards reduced molecular rates but without a corresponding slowdown in morphological evolution. Our study reveals that the living-fossil concept captures a quantifiable pattern of prolonged morphological conservatism, but also suggests that living fossils lack other shared biological or life-history characteristics that might unite them as a group.

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Comparing the Influence of Habitat Configuration on Population Connectivity and Genetic Structure Using Congeneric Species Across Multiple Taxa

Takenaka, M.; Okamoto, S.; Suzuki, H.; Yoshida, T.; Ueki, G.; Tojo, K.

2026-09-28 zoology 10.64898/2026.09.25.754572 medRxiv
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Abstract Habitat configuration influences population connectivity and, consequently, genetic structure. River networks provide heterogeneous, hierarchically arranged environments that shift drastically from upstream to downstream. We compared congeneric species across Ephemeroptera, Plecoptera, and Trichoptera, emphasizing longitudinal replacement (upstream to downstream) and the rarely studied wet rock (hygropetric) habitats. We surveyed six rivers on the Muroto Peninsula, Japan, qualitatively sampling aquatic insects at 63 sites. Cluster analysis based on the environmental data classified surveyed sites into four clusters. We analyzed a total of 10 species from four genera that exhibit longitudinal replacement patterns within genera. Genetic analyses based on the mitochondrial cytochrome c oxidase subunit I region revealed that upstream species showed higher genetic diversity than downstream species. In contrast, species adapted to hygropetric exhibited the lowest genetic differentiation among all habitat types. A novel contribution of this study is the inclusion of hygropetric species. The surprisingly low differentiation in hygropetric species suggests high connectivity, similar to lentic species. By comparing congeneric taxa across orders within environmentally similar rivers, we reduce phylogenetic and environmental confounds, strengthening inference that habitat configuration and dispersal traits jointly shape genetic structure. These findings provide a new perspective on riverine spatial ecology and underscore the importance of microhabitat-aware comparisons for evolutionary inference.

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Genomic and phenotypic consequences of long-term isolation and small population size in Yakushima macaques

Ito, T.; Hayakawa, T.; Kurihara, Y.; Hanya, G.; Kaneko, A.; Natsume, T.; Aisu, S.; Honda, T.; Wakamori, H.; Yachimori, S.; Anezaki, T.; Omi, T.; Hayama, S.-i.; Kawamoto, Y.

2026-10-04 evolutionary biology 10.64898/2026.10.02.752851 medRxiv
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The relative roles of genetic drift, inbreeding, and natural selection in island evolution have been fundamental themes in evolutionary biology for nearly a century. However, empirical studies have yet to reach a consensus, and research integrating both genotypes and phenotypes in primates remains limited. The present study aims to estimate the demographic history of an island primate population, the Yakushima macaque (Macaca fuscata yakui), and evaluate the genomic and morphological consequences of its isolation. To this end, we conducted population genetic analyses based on whole-genome sequencing and geometric morphometric analyses of cranial shape. Our results indicate that M. f. yakui has been isolated with an effective population size of several thousand individuals for over 100,000 years. Although we detected signatures of purifying selection, deleterious alleles have not been sufficiently purged. The degree of fluctuating asymmetry in males was slightly, yet significantly, higher in M. f. yakui than in mainland populations, whereas this trend was not observed in females. Marginal evidence of directional selection was found for cranial size, but not for shape. Overall, our findings suggest that due to long-term isolation, genetic drift and inbreeding have exerted a substantial influence on shaping genotypic and phenotypic variation in this island population.

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Revisiting Hydractiniidae (Hydrozoa): Mitogenomic Insights into the Diversity, Systematics and Life-Cycle Evolution of North Atlantic and Arctic Species

Beckmann, L. M.; Hosia, A.; Ronowicz, M.; Martell, L.; Soto-Angel, J. J.

2026-10-12 evolutionary biology 10.64898/2026.10.09.757902 medRxiv
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Hydractiniidae is a diverse family of colonial hydrozoans exhibiting substantial variation in colony organization, reproduction and life-cycle strategies, including species with and without a medusa stage. Several hydractiniids are also established model organisms, yet species boundaries and distributions remain incompletely resolved, limiting comparisons of reproductive and developmental traits across taxa. Here, we combine morphological and molecular data to reassess hydractiniid diversity, species identities, and life-cycle variation in material from the North Atlantic and Arctic Ocean. We integrate detailed observations of colonies and reproductive structures with DNA barcoding and mitochondrial genomes to evaluate species boundaries and provide molecular support for taxonomic identifications. Our results confirm 10 species in the study area, occurring from shallow waters to 2600 m depth and from coastal fjords to deep-sea hydrothermal vents. We further show that some historical species identifications and distribution records require reassessment. We establish a new genus for Arctic hydractiniid species (Bonneviactinia gen. nov.) and reassign two species to Schuchertinia. Mapping reproductive strategies onto the phylogeny reveals a broad pattern of increasing retention of medusa characters, from fixed sporosacs through reduced and fixed medusoids and non-liberated eumedusoids to fully developed, free-swimming feeding medusae, suggesting a progressive evolutionary trajectory in life-cycle complexity across Hydractiniidae. Furthermore, we clarify the identity of Podocoryna carnea and, through rearing experiments from polyp stage to medusae, characterize its morphological variation and developmental stages. By linking morphologically documented specimens and vouchers to genetic data, we establish reference resources that will facilitate future taxonomic, phylogenetic and biodiversity studies, including molecular surveys of marine communities. Together, these results demonstrate the value of integrative taxonomy for resolving hydractiniid diversity and provide a comparative foundation for investigating the evolution of reproductive strategies and life-cycle complexity in Medusozoa.

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What shapes intraspecific genetic diversity in amphibians and reptiles? A comparative macrogenetic study across six biogeographic regions

Paul, M. J.; Moreira, M. O.; Liz, A. V.; Decker, S. K.; Moritz, C.; Carstens, B. C.; Carvalho, S. B.

2026-10-02 evolutionary biology 10.64898/2026.09.29.755451 medRxiv
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Aim: Given that much of Earth's biodiversity is contained within species, inferring intraspecific genetic diversity and identifying its ecological correlates is essential for understanding evolutionary processes and informing conservation. Here, we investigate broad-scale variation in species-level mitochondrial genetic diversity and evaluate the relative importance of alternative ecological hypotheses proposed to explain it. Location: Brazilian Atlantic Forest, Iberian Peninsula, northwestern Africa, Arabian Peninsula, Madagascar and Australian Monsoonal Tropics. Taxon: Amphibians and reptiles Methods: We assembled a comprehensive dataset by integrating and manually curating mitochondrial sequences, occurrence records, environmental data and species traits from multiple open-access sources. We used multiple mitochondrial genes to estimate mean nucleotide diversity for each study species and quantified regional variation in genetic diversity. We then used random forests models to evaluate the relative importance of selected variables representing hypotheses related to water-energy availability, topography, historical climatic stability, land-cover characteristics, species geographic characteristics, life-history traits, evolutionary history and conservation status. Results: The final dataset comprised 247 amphibians and 295 reptiles, for which 43 and 52 explanatory variables were evaluated, respectively. Overall, the strongest support was found for hypotheses related to life-history traits, topographic heterogeneity and water-energy availability. In amphibians, higher genetic diversity was associated with smaller body size and greater environmental heterogeneity, particularly broader elevational and precipitation gradients. In reptiles, genetic diversity was primarily associated with life-history traits, with species of smaller body size and lower reproductive output exhibiting higher diversity. Long-term climatic stability showed comparatively little explanatory power for both taxa. Main conclusions: Our comprehensive study demonstrates that integrating extensive manual data curation with comparative macrogenetic analyses provides a robust framework for identifying broad-scale ecological correlates of mitochondrial genetic diversity. These findings improve our understanding of the factors associated with contemporary patterns of genetic diversity and provide a foundation for future conservation assessments in data-limited systems.

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Acrean fish diversity back in time: Paleoichthyology of the Rio Acre fossil fauna in Brazil

Ballen, G. A.; Alencar, K.; Muniz, F.; Guilherme, E.; Aldrin, L.; Lomba, S.; Dias-Costa, L. R.; Hsiou, A. S.

2026-10-02 paleontology 10.64898/2026.09.27.754785 medRxiv
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Freshwater fishes are an important component of the Neogene fossil record of South America. They have been recorded in the Acre fossil fauna, with considerable taxonomic and ecological diversity. Novel findings from the Patos locality in Acre State have produced several specimens representing eleven taxa, including the first occurrences in the fossil record of the genera Agamyxis, Glanidium, and Trachycorystes. These taxa comprise two orders, Characiformes and Siluriformes, with four and seven taxa respectively. The taxa found in the Patos locality are representative of typical modern Amazonian assemblages found in the drainages of the region, thus suggesting substantial ecological conservatism. An updated list of the Rio Acre fossil fish fauna is consistent with previous findings in the literature regarding clustering structure in fossil assemblages during the Neogene of South America.

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Unique pattern of injuries in Olenoides serratus from the Burgess Shale elucidate moulting process in trilobites

Losso, S. R.; Bicknell, R. D. C.

2026-09-24 paleontology 10.64898/2026.09.18.752702 medRxiv
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Ecdysozoans is one of the most abundant and diverse groups of metazoans and grow through moulting the rigid exoskeleton that provided protection. But the moulting process is often dangerous, resulting in malformations or death. Injuries have been well documented in the calcitic exoskeleton of trilobites, some of which have been attributed to moulting complications when long spines are deformed. Injuries in trilobites are often found in the trunk rather than the cephalon, and abnormal genal spines are rarely documented. Here we examine injuries in eleven specimens of Olenoides serratus from the Burgess Shale (Cambrian, Wuluian) and demonstrate an increased frequency of genal spine malformations compared to other trilobites. The unusual pattern of malformations may be attributed to moulting injuries of the long and delicate genal spines in O. serratus. To moult, trilobites curved ventrally to press the anterior margin of their cephalon into the substrate and open the facial sutures. This process exerted force on the librigenae. To exit the exuvia, the genal spines were forced to bend as the old librigenae dipped ventrally along the anterior edge, resulting in the injured genal spines in O. serratus. A similar process of moulting resulting in injuries may be seen in other early Euarthropods.

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Evolutionary history of aquaporin-10 pseudogenization in Cetartiodactyla

Nagai, N.; Nagashima, A.; Kato, A.

2026-09-24 evolutionary biology 10.64898/2026.09.20.752922 medRxiv
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Aquaporin-10 (AQP10) is an aquaglyceroporin that transports small, uncharged molecules, such as water and glycerol. In some species, such as mice and cattle, the AQP10 gene (Aqp10) is pseudogenized. In rodents, pseudogenization of Aqp10 is limited to the Myomorpha suborder, while Aqp10 remains intact in other rodent lineages. The order Cetartiodactyla encompasses diverse species, including Suina, Tylopoda, Ruminantia, and Whippomorpha. The distribution of intact Aqp10 across the entire Cetartiodactyla and analyses estimating the timing of Aqp10 pseudogenization have not been sufficiently investigated. In this study, we identified species within Cetartiodactyla that retain intact Aqp10 and analyzed the detailed patterns of pseudogenization to elucidate the specifics of pseudogenization within this order. We collected Aqp10 loci by BLAST and synteny analyses of publicly available genome databases of 51 cetartiodactyl species from 17 families. Dot plot and alignment analyses revealed that seudogenization of Aqp10 occurred in 42 Cetartiodactyla species, including the Camelidae, warthog (Suidae), Antilocapridae, okapi (Giraffidae), Cervidae, Bovidae, Hippopotamidae, and Cetacea. In contrast, nine species retained an intact Aqp10: pig and babirusa (Suidae), peccary (Tayassuidae), mouse deeres (Tragulidae), giraffes (Giraffidae), and musk deers (Moschidae). The timing of pseudogenization was estimated by comparing patterns of exon deletions, frameshifts, and nonsense mutations in Aqp10 pseudogenes. RT-PCR analysis of cattle and sheep tissues revealed that these species express Aqp3 and Aqp7 in their intestines, but not Aqp10. Collectively, these findings indicate that Aqp10 pseudogenization occurred independently in multiple cetartiodactyl lineages and provide new insights into the evolutionary history of Aqp10 loss within this order.

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Precipitation seasonality and soil carbon shape Arcellinida diversity across Bulgaria's ecosystems

Garcia Bodelon, A.; Todorov, M.; ElKhouri-Vidarte, N.; Soler-Zamora, C.; Fournier, B.; Lara, E.

2026-09-29 ecology 10.64898/2026.09.29.755277 medRxiv
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Recent years have witnessed the discovery of an immense diversity of protists in almost all ecosystems on Earth. However, a major challenge consists in identifying diversity patterns and the drivers that explain the distribution of this diversity. While several works have focused on local parameters, only a few studies have addressed this question at regional scales. In this study, we characterize the diversity and distribution of Arcellinida (testate amoebae) at regional scale of a climatically contrasted country, Bulgaria. We collected 75 samples geographically evenly distributed and representative of the country's natural ecosystems and used metabarcoding to determine their diversity. Communities composition differed markedly among habitats, revealing sharp beta diversity patterns. "Mesophilic" environments such as forests and ponds were most diverse, while acidic and oligotrophic Sphagnum peatlands were poorest and hosted the highest proportion of unique taxa. Our analyses of richness-environment based on a Random Forest approach applied to all soil systems indicated that stable humidity (low silt content, low precipitation seasonality) and intermediate soil carbon stocks had a positive effect on richness. The predicted increase in continentality in the next decades in Bulgaria calls for testing climatic models to evaluate the potential risk of declining Arcellinida biodiversity.

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Unexpected excess heterozygosity in a threatened, mycoheterotrophic, self-pollinating orchid

Barrett, C.; Fama, N. M.; Freudenstein, J. V.; Salazar, G.; Simon, S. J.; Sinn, B. T.

2026-10-09 evolutionary biology 10.64898/2026.10.07.757398 medRxiv
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Self-fertilization results in reduction of heterozygosity over time, accelerated by small population sizes, restricted ranges, and fragmented habitats. Yet some species maintain heterozygosity despite these factors. One is Corallorhiza bentleyi, a threatened, mycoheterotrophic, self-pollinating orchid restricted to Virginia and West Virginia, USA. Reduced representation sequencing was employed across five of the ~12 known localities of C. bentleyi, including representatives of sister species C. involuta from Mexico, to quantify patterns of population structure and genomic diversity. Surveys of C. bentleyi revealed a predominance of the cleistogamous (closed-flowered) growth form. Differentiation was observed between the two species but with weak population structure in C. bentleyi. Surprisingly, excess heterozygosity and negative inbreeding were recovered in both species, counter to expectations. Analyses provided evidence against bioinformatic artifacts (collapsed paralogs), small-sample bias (Robertson effect), and accumulation of mutations along chromosomes due to low recombination associated with selfing (Meselson effect) as explanations for excess heterozygosity. Demographic analyses favored a model of post-glacial divergence among C. bentleyi and C. involuta lacking bottlenecks. Finally, simulations under alternative selection scenarios favored models of overdominance vs. those of negative frequency dependent selection. It was concluded that overdominance, namely selection against selfed homozygotes, is the most likely explanation for elevated heterozygosity in C. bentleyi. Conservation concerns are discussed in light of these findings and the threats of extinction for one of the most vulnerable orchid species in North America.

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Scale-dependent associations between genetic similarity and aggregation behaviour in three toad lineages

Hase, K.; Abe, M. S.

2026-10-03 evolutionary biology 10.1101/453316 medRxiv
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Social aggregation is widespread in vertebrate larvae, yet how genetic relatedness shapes group cohesion remains poorly understood. In particular, the association between genetic similarity and social aggregation may depend on the scale at which genetic relatedness is evaluated. Here, we quantitatively examined aggregation behaviour in tadpoles from three lineages of Japanese toads, Bufo japonicus, B. formosus, and B. gargarizans miyakonis . We experimentally combined tadpole cohorts differing in relatedness and lineage and quantified aggregation using spatial dispersion analyses. We then examined relationships between aggregation behaviour and genetic variation using mitochondrial DNA (mtDNA), major histocompatibility complex (MHC) class II variation, and microsatellite diversity. Overall degree of aggregation did not differ significantly between sibling and non-sibling cohorts, and Morisitas index provided no evidence that tadpoles formed separate clumps according to relatedness. However, the correlation between genetic variation and aggregation differed across scales of genetic relatedness: among non-sibling cohorts, greater mtDNA pairwise distance was associated with weaker aggregation, whereas among sibling cohorts, greater MHC amino-acid difference was associated with weaker aggregation. These results indicate that the degree of aggregation is related to genetic similarity across different scales of relatedness, rather than being explained by a simple sibling/non-sibling distinction. Our findings suggest that tadpole group cohesion is related to genetic similarity in a scale-dependent manner, highlighting the importance of considering the scale of genetic relatedness when investigating social discrimination in vertebrates, while the sensory and molecular mechanisms underlying these relationships remain to be determined.

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Individual variation in glucocorticoid plasticity to temperature is not associated with reproductive success in a wild songbird population

Udino, E.; Deimel, C.; Hau, M.; Moiron, M.

2026-09-24 evolutionary biology 10.64898/2026.09.22.750903 medRxiv
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Vertebrates cope with rapid temperature fluctuations by plastically adjusting their behaviour and physiology. This phenotypic plasticity is partly mediated by plastic changes in glucocorticoid levels; however, whether individual differences in endocrine plasticity are associated with fitness remains unknown. Here, we examined the relationship between individual variation in glucocorticoid plasticity in response to air temperature changes and reproductive success in a wild great tit population (Parus major). Over nine years, we collected repeated measurements of baseline and stress-induced corticosterone during nestling rearing. We found consistent reaction norm patterns in hormone response to temperature variation -- as shown in a previous study conducted over a shorter period. This provides robust evidence of individual differences in average hormone levels (intercepts) and the strength of the plastic response (slopes), indicating potential for evolution. However, neither intercepts nor slopes were associated with reproductive success (fledgling number or brood mass), suggesting that individual variation in endocrine plasticity was not under natural selection, at least within the environmental conditions experienced. This study provides a first empirical test of the evolutionary implications of endocrine plasticity in the wild and highlights the need to assess its fitness relevance under more extreme conditions, such as those expected under future climate scenarios.

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Blackcaps (Sylvia atricapilla) with blood parasites have impaired oxidative status during autumn migration

Jimenez-Gallardo, L.; Lopez-Arrabe, J.; Perez-Tris, J.; Remacha, C.

2026-10-02 evolutionary biology 10.64898/2026.09.30.755661 medRxiv
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Parasites decrease host fitness, more so during physiologically challenging life stages such as migration. However, parasites are unevenly distributed among individuals, and hosts with different physiological condition may face different costs of migration with infections. We tested whether the relationship between infection by haemosporidian parasites and oxidative balance during autumn migration in blackcaps depends on individual age and sex, two major sources of variation in prevalence and physiological condition. We captured blackcaps at stopover in central Spain and measured infection status and physiological parameters. Blackcaps physiologically more prepared to depart from stopover had lower antioxidant capacity of plasma, probably because of physiological adjustments prior to the upcoming flight stage. Plasmodium parasites occurred only in coinfections, which were more frequent among young blackcaps. Regardless of individual sex and age, coinfected birds had lower antioxidant capacity of plasma, and blackcaps coinfected with Plasmodium had more oxidative damage to lipids in their red blood cells. Our results support the idea that haemosporidian infections impair the oxidative balance of blackcaps during autumn migration, which could impair individual performance and ultimately fitness, with increased population impacts during the first year of life when a larger proportion of individuals migrate coinfected with harmful Plasmodium parasites.