Global warming threatens species with temperature-dependent sex determination (TSD) by risking extreme offspring sex-ratio bias. In sea turtles, warmer incubation conditions produce more females. Such biases can transiently inflate apparent population growth before male scarcity undermines reproduction, possibly leading to population extinction. Here, we combine 15 years of field monitoring, drone-based sex-ratio surveys, Argos telemetry, and eco-physiological modelling to test whether rising temperatures underpin the rapid increase in loggerhead (Caretta caretta) nesting in Cabo Verde, one of the world largest populations. Historical air temperature, lagged by one generation, predicts current nest counts, consistent with a climate-driven, female-biased population. Drone surveys reveal an approximately 9:1 female-to-male breeding sex ratio across two consecutive breeding seasons, and simulations reproduce observed nesting trajectories under historical warming but not under null (no-warming) scenarios. Extending the analysis to 28 global loggerhead populations reveals consistent links between temperatures, latitude, and nesting trends. Our results identify warming-induced sex ratio bias as a key, yet deceptive, driver of apparent recovery, highlighting the need to reassess conservation success through the lens of demography of TSD species. ### Competing Interest Statement The authors have declared no competing interest. UK Research and Innovation Natural Environment Research Council, NE/V001469/1, NE/X012077/1 Queen Mary University of London, https://ror.org/026zzn846, QMUL Beasiswa Pendidikan Indonesia (Indonesian Education Scholarship, Center for Higher Education Funding and Assessment and Indonesian Endowment Fund for Education), FR202403000977 United States Fish and Wildlife Service, USFWS The Regional Partnership for Coastal and Marine Conservation in West Africa, PRCM

FRMPD2 is a human-specific multi-copy gene with higher mRNA expression in brain tissue, but its role in synapse formation and neurodevelopment remains unknown. We find that FRMPD2 is a neuron-specific protein with a high expression level in human brains compared to rodent brains. FRMPD2 overexpression in rat neurons stimulates synaptogenesis, leading to increased synaptic activities. Importantly, our results show that FRMPD2 via its PDZ domains recruits and enhances neuroligin-1 protein levels at the postsynaptic sites, and via its FERM domain interacts with the F-actin network in the spine. Increased expression of FRMPD2 also promotes spine formation and maturation, a foundational process for synapse formation. In developing embryonic mouse brains expressing higher FRMPD2 protein levels, we observed delayed neuronal migration, presumably promoting a protracted timeline for cortical lamination as a feature in human brain development. Behaviorally, mice with FRMPD2 overexpression in the brain demonstrate enhanced spatial memory retention. These findings indicate an important function for FRMPD2 in neuronal connectivity, brain development, and cognitive function.

Batesian mimics, such as hoverflies, can gain protection by resembling noxious species that predators avoid, such as bees. Such resemblance can be behavioural as well as morphological. Bees commonly have a preference for blue flowers, whilst flies generally prefer yellow and white. We predicted that, driven by selection for enhanced mimicry, bee-mimicking hoverflies would have switched flower preference from yellow to blue, to follow the flower choices of their models. We gathered internet photographs of flower-visiting bees and wasps, their hoverfly mimics, and non-mimicking hoverflies, and compared the colours of the visited flowers. Flowers visited by bumblebees were "bluer" than those visited by other hymenopterans (honeybees, wasps and solitary bees). Correspondingly, flowers visited by bumblebee-mimicking hoverflies were bluer than those visited by non-mimics, and were indistinguishable in blueness from those visited by bumblebees. There was no such pattern in flower "yellowness", where all flies and most hymenopterans preferred similar yellowness, and only bumblebees chose less yellow flowers than the others. Our study demonstrates widespread changes in microhabitat choice (specifically flower preference) among a diverse group of Batesian mimics, consistent with the idea of behavioural mimicry. Mildly noxious models such as bumblebees are thought to exert particularly strong selection for mimetic accuracy, suggesting that habitat-choice mimicry may be selected as a way of enhancing morphological mimicry. ### Competing Interest Statement The authors have declared no competing interest.

IntroductionRewilding, the facilitation of self-sustaining and resilient ecosystems by restoring natural processes, is an increasingly popular conservation approach and potential solution to the biodiversity and climate crises. Outcomes of rewilding can be unpredictable, and monitoring is essential to determine whether ecosystems are recovering. Metabarcoding, particularly of environmental DNA (eDNA), is revolutionizing biodiversity monitoring and could play an important role in understanding the impacts of rewilding but has mostly been applied within aquatic systems.MethodsThis systematic review focuses on the applications of eDNA metabarcoding in terrestrial monitoring, with additional insights from metabarcoding of bulk and ingested DNA. We examine publication trends, choice of sampling substrate and focal taxa, and investigate how well metabarcoding performs compared to other monitoring methods (e.g. camera trapping).ResultsTerrestrial ecosystems represented a small proportion of total papers, with forests the most studied system, soil and water the most popular substrates, and vertebrates the most targeted taxa. Most studies focused on measuring species richness, and few included analyzes of functional diversity. Greater species richness was found when using multiple substrates, but few studies took this approach. Metabarcoding did not consistently outperform other methods in terms of the number of vertebrate taxa detected, and this was likely influenced by choice of marker, sampling substrate and habitat.DiscussionOur findings indicate that metabarcoding, particularly of eDNA, has the potential to play a key role in the monitoring of terrestrial rewilding, but that further ground- truthing is needed to establish the most appropriate sampling and experimental pipelines for the target taxa and terrestrial system of interest.Systematic Review Registrationhttps://osf.io/38w9q/?view_only=47fdab224a7a43d298eccbe578f1fcf0, identifier 38w9q.

Behavioural variation among individuals is a hallmark of cooperative societies, which commonly contain breeders and non-breeders, helpers and non-helpers. In some cases, labour is divided with non-breeders "helping." Conversely, in some societies, subordinate non-breeders may not help. These individuals may be (i) an insurance workforce ensuring continuity of help for breeders when other helpers are lost, (ii) conserving energy while waiting to breed themselves or (iii) simply of too poor physiological quality either to help or breed. In the Australian Outback, Acacia thrips Dunatothrips aneurae (Thysanoptera) glue Acacia phyllodes into "domiciles" using silk-like secretions, either alone or cooperatively. Domicile maintenance is important for humidity, so repair can be interpreted as helping. I found that not all females helped to repair damage; some repaired partially or not at all ("non-helpers"). At the same time, some co-foundresses are non- or partially reproductive ("non-breeders"), and their role is currently unknown. I first tested the possibility that helping and breeding are divided, with non-helping females breeding, and non-breeders helping. In a lab experiment, I rejected this idea. Experimentally damaged domiciles were typically repaired by reproductive females, and less so by non- or partially reproductive individuals. To test whether non-helpers are an insurance workforce, I successively removed repairing females and found that non-helping females from the same domicile did not increase effort as a result, rejecting this hypothesis. Then I tested whether non-helpers were conserving energy waiting to breed. In a field experiment, I removed all other females, allowing either a helpful female or a non-helper to "inherit" her domicile. Isolated like this, non-helpers laid very few eggs compared to helpers or naturally occurring single foundresses, despite similar ovarian development. My findings show that labour was not divided: reproduction and helping covaried positively, probably depending on individual variation in female quality and intra-domicile competition. Non-helping females were neither an insurance workforce nor conserving energy waiting to breed. They are likely simply of poor quality, freeloading by benefiting from domicile maintenance by others. I hypothesize they are tolerated because of selection for indiscriminate communal brood care in the form of domicile repair.

Genetic screens in cancer cell lines inform gene function and drug discovery. More comprehensive screen datasets with multi-omics data are needed to enhance opportunities to functionally map genetic vulnerabilities. Here, we construct a second-generation map of cancer dependencies by annotating 930 cancer cell lines with multi-omic data and analyze relationships between molecular markers and cancer dependencies derived from CRISPR-Cas9 screens. We identify dependency-associated gene expression markers beyond driver genes, and observe many gene addiction relationships driven by gain of function rather than synthetic lethal effects. By combining clinically informed dependency-marker associations with protein-protein interaction networks, we identify 370 anti-cancer priority targets for 27 cancer types, many of which have network-based evidence of a functional link with a marker in a cancer type. Mapping these targets to sequenced tumor cohorts identifies tractable targets in different cancer types. This target prioritization map enhances understanding of gene dependencies and identifies candidate anti-cancer targets for drug development.

A reduction in AMPA receptor (AMPAR) expression and weakened synaptic activity is early cellular phenotypes in Alzheimer’s disease (AD). However, the molecular processes leading to AMPAR downregulation are complex and remain less clear. Here, we report that the salt inducible kinase SIK1 interacts with AMPARs, leading to a reduced accumulation of AMPARs at synapses. SIK1 protein level is sensitive to amyloid beta (Aβ) and shows a marked increase in the presence of Aβ and in AD brains. In neurons, Aβ incubation causes redistribution of SIK1 to synaptic sites and enhances SIK1-GluA1 association. SIK1 function is required for Aβ-induced AMPAR reduction. Importantly, in 3xTG AD mice, knockdown of SIK1 in the brain leads to restoration of AMPAR expression and a rescue of the cognitive deficits. These findings indicate an important role for SIK1 in meditating the cellular and functional pathology in AD.

Background: Characterising genetic and epigenetic diversity is crucial for assessing the adaptive potential of populations and species. Slow-reproducing and already threatened species, including endangered sea turtles, are particularly at risk. Those species with temperature-dependent sex determination (TSD) have heightened climate vulnerability, with sea turtle populations facing feminisation and extinction under future climate change. High-quality genomic and epigenomic resources will therefore support conservation efforts for these flagship species with such plastic traits. Findings: We generated a chromosome-level genome assembly for the loggerhead sea turtle (Caretta caretta) from the globally important Cabo Verde rookery. Using Oxford Nanopore Technology (ONT) and Illumina reads followed by homology-guided scaffolding, we achieved a contiguous (N50: 129.7 Mbp) and complete (BUSCO: 97.1%) assembly, with 98.9% of the genome scaffolded into 28 chromosomes and 29,883 annotated genes. We then extracted the ONT-derived methylome and validated it via whole genome bisulfite sequencing of ten loggerheads from the same population. Applying our novel resources, we reconstructed population size fluctuations and matched them with major climatic events and niche availability. We identified microchromosomes as key regions for monitoring genetic diversity and epigenetic flexibility. Isolating 191 TSD-linked genes, we further built the largest network of functional associations and methylation patterns for sea turtles to date. Conclusions: We present a high-quality loggerhead sea turtle genome and methylome from the globally significant East Atlantic population. By leveraging ONT sequencing to create genomic and epigenomic resources simultaneously, we showcase this dual strategy for driving conservation insights into endangered sea turtles. ### Competing Interest Statement The authors have declared no competing interest.

ABSTRACT To date, studies of the impacts of climate warming on individuals and populations have mostly focused on mortality and thermal tolerance. In contrast, much less is known about the consequences of sublethal effects, which are more challenging to detect, particularly in wild species with cryptic life histories. This necessitates the development of molecular tools to identify their signatures. In a split‐clutch field experiment, we relocated clutches of wild, nesting loggerhead sea turtles (Caretta caretta) to an in situ hatchery. Eggs were then split into two sub‐clutches and incubated under shallow or deep conditions, with those in the shallow treatment experiencing significantly higher temperatures in otherwise natural conditions. Although no difference in hatching success was observed between treatments, hatchlings from the shallow, warmer treatment had different length–mass relationships and were weaker at locomotion tests than their siblings incubated in the deep, cooler treatment. To characterise the molecular signatures of these thermal effects, we performed whole genome bisulfite sequencing on blood samples collected upon emergence. We identified 287 differentially methylated sites between hatchlings from different treatments, including on genes with neurodevelopmental, cytoskeletal, and lipid metabolism functions. Taken together, our results show that higher incubation temperatures induce sublethal effects in hatchlings, which are reflected in their DNA methylation status at identified sites. These sites could be used as biomarkers of thermal stress, especially if they are retained across life stages. Overall, this study suggests that global warming reduces hatchling fitness, which has implications for dispersal capacity and ultimately a population's adaptive potential. Conservation efforts for these endangered species and similar climate‐threatened taxa will therefore benefit from strategies for monitoring and mitigating exposure to temperatures that induce sublethal effects.

The naked mole-rat Heterocephalus glaber is a eusocial mammal exhibiting extreme longevity (37-year lifespan), extraordinary resistance to hypoxia and absence of cardiovascular disease. To identify the mechanisms behind these exceptional traits, metabolomics and RNAseq of cardiac tissue from naked mole-rats was compared to other African mole-rat genera (Cape, Cape dune, Common, Natal, Mahali, Highveld and Damaraland mole-rats) and evolutionarily divergent mammals (Hottentot golden mole and C57/BL6 mouse). We identify metabolic and genetic adaptations unique to naked mole-rats including elevated glycogen, thus enabling glycolytic ATP generation during cardiac ischemia. Elevated normoxic expression of HIF-1α is observed while downstream hypoxia responsive-genes are down-regulated, suggesting adaptation to low oxygen environments. Naked mole-rat hearts show reduced succinate levels during ischemia compared to C57/BL6 mouse and negligible tissue damage following ischemia-reperfusion injury. These evolutionary traits reflect adaptation to a unique hypoxic and eusocial lifestyle that collectively may contribute to their longevity and health span.

Climate change and climate variability are affecting marine mammal species and these impacts are projected to continue in the coming decades. Vulnerability assessments provide a framework for evaluating climate impacts over a broad range of species using currently available information. We conducted a trait-based climate vulnerability assessment using expert elicitation for 108 marine mammal stocks and stock groups in the western North Atlantic, Gulf of Mexico, and Caribbean Sea. Our approach combined the exposure (projected change in environmental conditions) and sensitivity (ability to tolerate and adapt to changing conditions) of marine mammal stocks to estimate vulnerability to climate change, and categorize stocks with a vulnerability index. The climate vulnerability score was very high for 44% (n = 47) of these stocks, high for 29% (n = 31), moderate for 20% (n = 22), and low for 7% (n = 8). The majority of stocks (n = 78; 72%) scored very high exposure, whereas 24% (n = 26) scored high, and 4% (n = 4) scored moderate. The sensitivity score was very high for 33% (n = 36) of these stocks, high for 18% (n = 19), moderate for 34% (n = 37), and low for 15% (n = 16). Vulnerability results were summarized for stocks in five taxonomic groups: pinnipeds (n = 4; 25% high, 75% moderate), mysticetes (n = 7; 29% very high, 57% high, 14% moderate), ziphiids (n = 8; 13% very high, 50% high, 38% moderate), delphinids (n = 84; 52% very high, 23% high, 15% moderate, 10% low), and other odontocetes (n = 5; 60% high, 40% moderate). Factors including temperature, ocean pH, and dissolved oxygen were the primary drivers of high climate exposure, with effects mediated through prey and habitat parameters. We quantified sources of uncertainty by bootstrapping vulnerability scores, conducting leave-one-out analyses of individual attributes and individual scorers, and through scoring data quality for each attribute. These results provide information for researchers, managers, and the public on marine mammal responses to climate change to enhance the development of more effective marine mammal management, restoration, and conservation activities that address current and future environmental variation and biological responses due to climate change.

Same-sex sexual behaviours (SSBs) are widespread across the animal kingdom, yet little consensus exists regarding their potential adaptive functions or evolutionary history. To fully address questions such as why or how a trait has evolved, it is important to first understand its natural history. Here, we present evidence of SSBs in two male naked mole-rats for the first time. Due to extreme natal philopatry, most naked mole-rats never disperse, making encounters with potential mates valuable in fitness terms. However, in captive colonies of naked mole-rats, a rare disperser morph has been described. We speculate that such dispersing individuals may be indiscriminatory in their mating strategy, copulating with conspecifics regardless of sex, to counter the risk of missed reproductive opportunities. This behavioural strategy would contrast with the common characterisation of naked mole-rats, which describes them as fiercely xenophobic and hostile to unfamiliar individuals. Further, the observation of an indiscriminate mating strategy in a species with limited opportunities for dispersal provides interesting context for the study of the evolution of SSBs.

Selective serotonin reuptake inhibitors (SSRIs) are commonly prescribed drugs for treating human depression, but the role of serotonin—often called the happy chemical—is complex, associated with the regulation of appetite, anxiety, aggression, and more. With such psychoactive pharmaceuticals being increasingly detected in aquatic environments and their effects on non-target species an uncertainty, this study seeks to investigate how inhibiting the serotonin pathway with the SSRI fluoxetine affects territorial aggression, using the cichlid fish Nyassachromis cf. microcephalus as a model. Males of this sand-dwelling Lake Malawi species build bowers to attract females within a lekking system, where male-male competition is intense. Being aggressive in defending against intruders would serve to maximise mating opportunities and reproductive success for the territory holding male. However, after a one-week exposure to fluoxetine at environmentally relevant concentrations, a decline in aggression was observed in the male cichlids. This implies the serotonergic system plays an important role in modulating aggression and therefore sexual selection in cichlid fishes, and that environmental pollution has the potential to disrupt these behaviours.### Competing Interest StatementThe authors have declared no competing interest.

Despite decades of research into the evolutionary drivers of sociality, we know relatively little about the underlying proximate mechanisms. Here we investigate the potential role of prolactin in the highly social naked mole-rat. Naked mole-rats live in large social groups but, only a small number of individuals reproduce. The remaining non-breeders are reproductively suppressed and contribute to burrow maintenance, foraging, and allo-parental care. Prolactin has well-documented links with reproductive timing and parental behaviour, and the discovery that non-breeding naked mole-rats have unusually high prolactin levels has led to the suggestion that prolactin may help maintain naked mole-rat sociality. To test this idea, we investigated whether urinary prolactin was correlated with cooperative behaviour and aggression. We then administered the prolactin-suppressing drug Cabergoline to eight female non-breeders for eight weeks and assessed the physiology and behaviour of the animals relative to controls. Contrary to the mammalian norm, and supporting previous findings for plasma, we found non-breeders had elevated urinary prolactin concentrations that were similar to breeding females. Further, prolactin levels were higher in heavier, socially dominant non-breeders. Urinary prolactin concentrations did not explain variation in working behaviour or patterns of aggression. Furthermore, females receiving Cabergoline did not show any behavioural or hormonal (progesterone) differences, and urinary prolactin did not appear to be suppressed in individuals receiving Cabergoline. While the results add to the relatively limited literature experimentally manipulating prolactin to investigate its role in reproduction and behaviour, they fail to explain why prolactin levels are high in non-breeding naked mole-rats, or how female non-breeding phenotypes are maintained.

1. Most organisms must regulate their nutritional intake in an environment full of complex food choices. While this process is well-understood for self-sufficient organisms, dependent offspring, such as bee larvae, in practice have limited food choices because food is provided by parents. Nutrient balancing may therefore be achieved by offspring, by parents on offspring's behalf, or by both, whether cooperatively or in conflict. 2. We used the Geometric Framework to investigate the capacity of dependent larval mason bees Osmia bicornis to regulate their intake of protein and carbohydrate, as well as these nutrients' effects upon growth and survival. Female Osmia seal eggs individually inside cells they have provisioned with pollen, and have no contact with developing offspring, allowing offspring choices to be studied in isolation. Herbivorous insect larvae are typically expected to balance protein and carbohydrate to maximize growth and reproduction. 3. Contrary to prediction, carbohydrate and not protein mediated both growth and survival to pupation. Accordingly, larvae prioritized maintaining a constant intake of carbohydrate and self-selected a relatively carbohydrate-biased diet compared to other hymenopterans, while tolerating wide excesses and deficiencies of protein, rendering them potentially vulnerable to dietary change or manipulation. Reasons for prioritizing carbohydrate may include (a) the relative abundance of protein in their normal pollen diet, (b) the relative paucity of nectar in parental provisions making carbohydrate a scarce resource or (c) the requirement for diapause for all O. bicornis larvae. Larvae were intolerant of moderate dietary dilution, likely reflecting an evolutionary history of nutrient-dense food. 4. Our results demonstrate that dependent offspring can remain active participants in balancing their own nutrients even when sedentary, and, moreover, even in mass provisioning systems where parents and offspring have no physical contact. Research should now focus on whether and how evolutionary interests of parent and dependent offspring coincide or conflict with respect to food composition, and the implications for species' resilience to changing environments.

Environmental DNA (eDNA) is one of the fastest developing tools for species biomonitoring and ecological research. However, despite substantial interest from research, commercial and regulatory sectors, it has remained primarily a tool for aquatic systems with a small amount of work in substances such as soil, snow and rain. Here we demonstrate that eDNA can be collected from air and used to identify mammals. Our proof of concept successfully demonstrated that eDNA sampled from air contained mixed templates which reflect the species known to be present within a confined space and that this material can be accessed using existing sampling methods. We anticipate this demonstration will initiate a much larger research programme in terrestrial airDNA sampling and that this may rapidly advance biomonitoring approaches. Lastly, we outline these and potential related applications we expect to benefit from this development.

BACKGROUND:The naked mole-rat (Heterocephalus glaber) is among the most social mammals on the planet, living in eusocial groups of up to 300 individuals that contain a single reproductive female and up to three reproductive males. A critical aspect of their complex social system is the division of labour that allows non-breeders to form an effective workforce. Age- or weight-based polyethisms are widely cited as explanations for how labour is divided, but evidence in support of these hypotheses has been equivocal.METHODS:To assess the extent to which individual working behaviour is determined by sex, age, weight and social rank, we studied the behaviours of 103 animals from eight captive colonies. We performed focal sampling and ran mixed-effects models to assess which factors explained variation in working behaviour during six ten-minute observation periods per individual.RESULTS:Contrary to widely-held beliefs, we found that working behaviour did not decrease linearly with weight, although polynomial regressions indicated younger and medium-sized individuals worked most frequently, while high-ranking individuals worked for the shortest periods of time. Working behaviour and its relationship with individual characteristics also varied between colonies.CONCLUSIONS:While age- or size-based polyethisms may have some influence on working behaviour, we argue that other characteristics of the individual and colony are also important. In particular, the interactions of individual, social and environmental factors must be considered in order to understand the emergence and effectiveness of the division of labour that is so critical to many social organisms.

Nutritional studies often require precise control of nutrients via dilution of artificial diets with indigestible material, but such studies in bees are limited. Common diluents like cellulose typically result in total mortality of bee larvae, making quantitative studies difficult. We investigated potential alternative dietary dilution agents, sporopollenin (pollen exines) and agar. We reared Osmia bicornis larvae on pollen diluted with these substances, alongside undiluted controls. Sporopollenin neither prevented nor improved survival, suggesting it is a suitable diluent. Agar appeared marginally to increase survival and its suitability requires further research. Both substances reduced cocoon weight, and sporopollenin also prolonged development, suggesting processing costs. Determining the physiological mechanisms driving these responses requires further work. Our findings should facilitate studies involving nutritional manipulations for solitary bees.