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Journal of Thermal Biology

Elsevier BV

Preprints posted in the last 30 days, ranked by how well they match Journal of Thermal Biology's content profile, based on 15 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.

1
Carryover effects modulate spring phenological responses to temperature in a herbivorous insect

Rattigan, S. D.; Beaupere, L. C.; Sheldon, B. C.; Learmonth, R.

2026-04-03 ecology 10.64898/2026.04.01.715835 medRxiv
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O_LIPhenological shifts are a major ecological consequence of climate change, yet studies often focus on single life stages meaning that the potential for carryover effects between life stages remains poorly understood. Failing to account for these effects may lead to inaccurate estimates of phenological shifts, with consequences for predicted synchrony among interacting species. This is especially relevant for temperate systems where climate warming is occurring unevenly across the year. C_LIO_LIHere, we investigated how temperature experienced the previous autumn and winter (during the pupal and egg stage) influences spring phenology in the winter moth (Operophtera brumata), a herbivorous insect with distinct life stages. Using 50 years of local climate data to create five experimental temperature regimes, we first quantified phenotypic plasticity in the duration and temporal variability of pupal and egg development. We then examined how timing of adult moth emergence affects timing of offspring hatching. C_LIO_LIWe found divergent effects of temperature on different life stages; pupal development time was shortest at intermediate temperatures while egg development time decreased linearly with increasing temperature. Furthermore, phenological shifts due to the conditions experienced by the mother were carried over to influence the phenology of her offspring. While this carryover effect was partially compensated during subsequent stages, compensation decreased under warming conditions. C_LIO_LIThese results refine our understanding of the sensitivity of the annual cycle of winter moth phenology to variation in temperature with potential implications for population dynamics and interspecific interactions. Overall, our findings highlight the need to consider the impacts of warming across multiple life stages so that carryover effects can be properly accounted for. Doing so will improve predictions of phenological shifts under future climates. C_LI

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A Fluorescent Dauer Marker in Caenorhabditis inopinata Enables Comparative Analysis of Dauer-Inducing Mechanisms

Iitsuka, R.; Haruta, N.; Oomura, S.; Sugimoto, A.

2026-04-09 developmental biology 10.64898/2026.04.06.716796 medRxiv
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Dauer larvae are a dormant developmental stage in nematodes that is induced by a range of environmental cues. The molecular mechanisms that transduce these cues to regulate dauer entry have been well characterized in Caenorhabditis elegans, whereas those in other nematode species remain unclear. The closest known sibling species of C. elegans, Caenorhabditis inopinata, occupies a distinct ecological niche and shows an extremely low frequency of dauer formation by starvation in laboratory conditions, suggesting that it could serve as a useful comparative model for analyzing dauer-inducing mechanisms. To support such analysis, we generated a fluorescent dauer reporter, Cin-col-183p::mCherry, in C. inopinata based on a previously reported dauer-specific reporter in C. elegans. This reporter showed fluorescence specifically in the pre-dauer and dauer stages, but not in other developmental stages, indicating that it functions as a dauer-specific marker in C. inopinata. Using these marker strains, we compared the responses to high temperature and RNAi-mediated knockdown of insulin/IGF-1 pathway genes (daf-2, age-1, and pdk-1), and found that dauer induction differs mechanistically between C. elegans and C. inopinata. This dauer-specific fluorescent strain will be a useful tool for investigating the diversity of dauer-inducing mechanisms across nematode species. Article SummaryDauer is a dormant developmental stage in nematodes induced by environmental stress. Although its regulation is well studied in Caenorhabditis elegans, the mechanisms in other species remain unclear. Here, we developed a fluorescent dauer reporter, Cin-col-183p::mCherry, in Caenorhabditis inopinata, a close relative of C. elegans. The reporter was specifically expressed in pre-dauer and dauer stages, confirming its usefulness as a dauer marker. Using this strain, we found that responses to high temperature and insulin/IGF-1 pathway gene knockdown differ between C. elegans and C. inopinata. This reporter will help reveal diversity in dauer-inducing mechanisms across nematode species.

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From Past to Future: The Impact of Climate Change on a Mediterranean Lizard

Kuyucu, A. C.

2026-03-27 ecology 10.64898/2026.03.26.714419 medRxiv
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Mediterranean Basin, one of the most important hot spots for reptiles, is also expected to experience significant impacts with climate change, posing a severe risk for the herpetofauna of the region. This study uses the snake-eyed lizard Ophisops elegans as a model organism to investigate the potential impacts of past and future climate change on reptile distributions in the region. An ecological niche model (ENM) was developed with the Maxent algorithm, with location points from GBIF and bioclimatic variables from the CHELSA dataset, then projected onto past LGM ([~]21 kya) and future (2071-2100 SSP3-7.0 and SSP5-8.5) scenarios. Results show that the present-day distribution of O. elegans is primarily driven by temperature seasonality and precipitation, indicating a preference for coastal Mediterranean climates with dry summers. The LGM projection suggests a fragmented and contracted range, confined to coastal refugia around the Mediterranean and Caspian Seas. Future projections for 2071-2100 show consistent and alarming contraction of suitable habitats under both SSP scenarios. In conclusion these findings indicate that O. elegans is vulnerable to significant habitat loss under projected climate change. This severe impact on a wide-spread species implies that the herpetofauna of the Mediterranean Basin may face a significant threat in future.

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No evidence of sentinel behaviour in a highly social bird based on an artificial set-up

Marmelo, M.; Silva, L.; Ferreira, A.; Doutrelant, C.; Covas, R.

2026-03-24 animal behavior and cognition 10.64898/2026.03.17.712373 medRxiv
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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.

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A capture-handling-restraint protocol elicits short-term stress responses in female and male parental wild great tits (Parus major) but has little impact on reproductive success

Frohlich, F.; Mentesana, L.; Deimel, C.; Hau, M.

2026-03-19 physiology 10.64898/2026.03.17.712382 medRxiv
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Capturing and handling wild animals is essential for ecological and evolutionary research, yet their effects on physiology, behaviour, and reproductive success remain poorly understood. We investigated short- and longer-term consequences of a capture-handling-restraint protocol in wild great tits (Parus major) over three breeding seasons. To assess short-term responses, we measured circulating corticosterone, a metabolic hormone that responds to unpredictable challenges, and automatically recorded provisioning behaviour. We also explored whether environmental and individual traits were related to provisioning latency (i.e., time to resume provisioning after capture). To evaluate longer-term effects, we monitored provisioning in the days following capture and related it to reproductive success (fledgling number and body condition). We predicted that longer handling would increase stress-induced corticosterone and provisioning latency, that these variables would be positively correlated, and that higher corticosterone and longer latencies would be associated with lower reproductive success. After capture, great tits showed elevated corticosterone and delayed provisioning. Contrary to our predictions, handling duration was negatively associated with stress-induced corticosterone in males (but not females) and did not affect provisioning latency. Provisioning latency was unrelated to corticosterone, environmental, or individual variables. Following capture, parents resumed provisioning, and short-term responses had little influence on reproductive success. We show that parental behaviour and physiology are affected by capture restraint protocols on the short term, but offspring condition and survival are not. However, these results should be interpreted cautiously, as our study lacks an uncaptured control group. Our findings highlight that evaluating welfare impacts requires rigorous study design incorporating both immediate and longer-term behavioural and fitness effects.

6
Can tropical agamid lizards physiologically tolerate the altered thermal mosaic of urban microhabitats?

Razak, M.; Ben, A.; Dhere, S.; Thaker, M.

2026-04-08 ecology 10.64898/2026.04.06.716463 medRxiv
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Urbanization and human-induced environmental changes create unique and unprecedented thermal landscapes, yet the extent to which species respond to these changes remains poorly understood. One major challenge in studying these responses is the spatial mismatch between the small scale at which organisms experience their environment and the broader scale at which climate data are typically collected. We use Infrared Thermography (IRT) to quantify the fine scale microclimate in urban and rural habitats used by two tropical agamid lizards, Calotes versicolor and Psammophilus dorsalis. By combining field-based body temperatures and lab-based measures of thermal limits (CTmax, CTmin)and preferences (Tpref), we assess how the thermal heterogeneity of these fine mosaics of microhabitats influence the degree of thermoregulation (k) of these species. We find that thermal responses to urbanization are shaped by species-specific thermal traits and patterns of microhabitat use. Between the species, urban individuals did not differ markedly in habitat thermal heterogeneity, substrate temperature used or degree of thermoconformity. However, within species, P. dorsalis experiences warmer and more heterogeneous conditions in rural habitats, whereas C. versicolor experiences similar thermal conditions across habitats. Calotes versicolor also exhibits broader thermal tolerance and preferred temperature ranges than P. dorsalis. Collectively, our results suggest that P. dorsalis may be more susceptible to the thermal constraints imposed by human-modified landscapes. Overall, we demonstrate the critical need to account for microclimatic conditions and species-specific thermal traits when determining how animals respond to changes in the thermal environment expected from climate change.

7
Multidecadal changes in land cover across a disturbance gradient in mountain grasslands of Kyrgyzstan

Young, S. C. E.; Watkins, H. V.; Brownlee, S. F.; Yan, H. F.; Cote, I. M.

2026-03-27 ecology 10.64898/2026.03.24.712710 medRxiv
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Mountain ecosystems face unprecedented pressures from anthropogenic activities and climate change, challenging the productivity of these vital habitats. In the Tien Shan mountains, understanding localized responses to these pressures is often hindered by the coarse spatiotemporal resolutions of available data. To address this, we combined high-resolution satellite imagery (1997-2021) to map land-cover dynamics in the Naryn oblast, Kyrgyzstan across a gradient of grazing intensities. We classified and quantified land-cover distribution over 24 years, investigating the roles of topography, elevation, and anthropogenic disturbances as drivers of change. Our results identify intermediate elevations, high degrees of disturbance, and the interaction between the two as the primary contributors to recent transitions in grassland, forest, and barren habitats. By integrating Landsat analysis-ready data, European Space Agency WorldCover dataset and digital elevation models at fine spatial scales, we provide valuable contemporary and historical landscape and habitat-level insights and a high-resolution framework for disentangling climate-driven shifts from land-use impacts. These findings highlight the urgency of localized management in remote, data-poor regions where rapid environmental change threatens both biodiversity and pastoral livelihoods. Our work serves as a critical baseline for characterizing the adaptability of semi-arid mountain rangelands under escalating global and regional pressures.

8
Resource abundance and dietary specialization predict elevational migration in a hyperdiverse montane bird community

Menon, T.; Tyagi, A.; Managave, S.; Ramakrishnan, U.; Srinivasan, U.

2026-03-20 ecology 10.64898/2026.03.18.710293 medRxiv
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Migration is a well-described behavioural strategy that allows species to track variation in resources and climatic conditions by moving in response to seasonality. A common form is elevational migration, an annual short-distance movement undertaken by many mountain bird species globally. While studies show that the timing of migration may relate to food availability, the mechanisms determining which species migrate remain unclear. Our study investigated if the degree of dietary specialization explains why some high-elevation bird species in seasonal environments migrate downslope for the winter while others remain resident at high altitudes despite the apparent scarcity of their preferred food resources. We mist-netted birds along a 2300-m elevational gradient in the Eastern Himalaya and collected blood and faecal samples from 261 individual birds belonging to 18 species of high-elevation residents (ten) and elevational migrants (eight) in their breeding and wintering ranges. Using stable isotope ratios of carbon and nitrogen in whole blood and faecal DNA metabarcoding, we compared their seasonal trophic levels and dietary niches. Nitrogen isotope ratios showed that residents had a substantially lower trophic position in the winter compared to summer (-0.35 [-0.52, -0.17]), whereas migrants had a slightly higher trophic position in the winter (0.15 [-0.02, 0.32]). This trophic shift in residents was likely due to a decrease in insectivory and an increase in frugivory in the winter. The frequency of key insect orders (Lepidoptera, Hemiptera, and Coleoptera) declined by 20-35% in their winter diets alongside an increase in fruit, particularly from the family Polygonaceae (0.33 [0.18, 0.46]). Additionally, compared with residents, migrants showed greater overlap in their dietary niches between summer and winter (98% vs 80%). Because arthropod abundances in the Himalayas peak at high elevations in the summer and decline in the winter, we suggest that elevational migrants are likely dietary specialists that track resources, while high-elevation residents are dietary generalists that supplement their winter diet with fruit and nectar because of the scarcity of arthropods. These findings indicate that a species dietary specialization is linked to its migratory behaviour, providing a potential mechanistic explanation for how different species solve the challenge of seasonal resource limitation.

9
Diet and breeding productivity in European Shag (Gulosus aristotelis): insights from two Portuguese colonies

Vieira, B.; Goncalves, D.; Oliveira, N.

2026-03-30 ecology 10.64898/2026.03.29.715095 medRxiv
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Climate change and anthropogenic pressures are reshaping marine food webs, altering prey availability and affecting top predators. The European Shag (Gulosus aristotelis), a coastal demersal seabird, provides a valuable model for examining environmentally mediated dietary variation, given its trophic plasticity and capacity to adjust prey use according to local availability, while also allowing assessment of potential demographic consequences. This study investigated spatial and temporal variation in diet at two Portuguese colonies (Berlengas and Arrabida) between 2016 and 2024 and assessed long-term reproductive productivity at Berlengas. A total of 468 regurgitated pellets were analysed, and diet composition was quantified using the Index of Relative Importance (IRI). Generalised additive models were applied to assess environmental, spatial, and period-specific effects on diet composition, while reproductive productivity was modelled in relation to prey biomass. Diet variation was primarily explained by environmental predictors, including sea surface temperature, chlorophyll-a concentration, and zooplankton, whereas year per se had no significant effect, indicating environmentally mediated bottom-up effects. Spatial differences between colonies reflected contrasting prey field structures, and period-specific patterns suggested increased specialisation during breeding. Higher biomass of sandeels (Ammodytidae) was positively associated with reproductive output, whereas shifts toward lower-energy prey were associated with reduced productivity. These findings demonstrate that environmentally driven dietary change has measurable demographic consequences, underscoring the importance of bottom-up processes in shaping seabird population dynamics and informing conservation strategies under ongoing climate change.

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Environmental factors that impact the development of infective juveniles of entomopathogenic nematode Steinernema hermaphroditum

Cao, M.

2026-04-08 developmental biology 10.64898/2026.04.07.717109 medRxiv
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Animals sense and integrate complex external cues to make developmental decisions that help them better survive and adapt to their natural habitats. Under environmental adversity, nematodes can enter an alternative developmental pathway to form a diapautic and stress-resistant stage, termed the dauer larvae. While dauer formation has been well characterized in Caenorhabditis elegans, how environmental factors influence analogous stages in other nematode species remains largely unexplored. This study examines how symbiotic bacteria, temperature, and pheromones affect the formation of the infective juvenile (IJ), a dauer-like stage, of the insect-parasitic nematode Steinernema hermaphroditum. In contrast to C. elegans, where dauer entry is promoted by heat, IJ development in S. hermaphroditum development is enhanced by reduced temperature. Moreover, the presence and absence of live symbiotic bacterium Xenorhabdus griffiniae functions as an ON-and-OFF switch that regulates the host IJ formation. Crude pheromone extracts from S. hermaphroditum liquid culture do not robustly induce IJ formation in a dose-responsive manner, unlike the potent pheromone-driven dauer entry observed in C. elegans. Nutrient-rich liver-kidney media that mimics host insect environment showed IJ entry induction in a pheromone-dependent manner. These data suggest that external cues, such as temperature, microbial diet, and pheromone, are perceived differently by S. hermaphroditum in comparison to that of C. elegans, reflecting species-specific adaptations to distinct ecological niches and life history strategies.

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Reassessing display behavior from Bels et al. (2025) given the complexity of anthropogenic hybridization and intraspecific diversity in Iguana iguana

van den Burg, M. P.; Thibaudier, J.

2026-03-23 zoology 10.64898/2026.03.19.713079 medRxiv
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Understanding behavioral differences between non-native and closely related endangered species could be important to aid conservation management. In volume 169 of Zoology, Bels et al. (2025) reported on their comparison of display-action-patterns (DAP) between native Iguana delicatissima and non-native iguanas present on islands of the Guadeloupe Archipelago in the Caribbean Lesser Antilles. Here, we address conceptual and methodological concerns about their work and reanalyze their data given our proposed corrections, primarily a literature-informed adjustment of their "species" category. We additionally utilize online videos from South American mainland I. iguana populations, from where the non-native iguanas in the Guadeloupe Archipelago originate, to better understand the different DAPs between native and non-native iguanas in the Guadeloupe Archipelago. Significant differences in DAP characteristics among "species" categories (native I. delicatissima, non-native iguanas, and hybrids) show that Bels et al. (2025) oversimplified their data analyses by merging all non-native populations into one group. This result indicates the presence of behavioral variation among subpopulations within widely hybridizing iguanid populations, which has been poorly studied. Additionally, videos from mainland populations across two major mitochondrial clades of Iguana iguana show that non-native iguanas on Guadeloupe retained DAP characteristics of those populations from which they originate. We discuss these findings in light of the proposed hypotheses put forward by Bels et al. (2025), of which two can be excluded. Overall, our reanalysis shows that studies focusing on characteristics within settings of complex hybridization in diverse species should acknowledge this complexity.

12
Wind pattern oscillations explain seabird movements at-sea: a nested multiscale approach

ROY, A.; Delord, K. C.; BARBRAUD, C.; TERRAY, P.

2026-04-03 ecology 10.64898/2026.04.01.715798 medRxiv
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Wind has a strong influence on the flight characteristics, movements, energetics, demography, life-history traits and biogeography of flying animals. With climate change affecting atmospheric circulation patterns at different time scales, understanding the links between wind and animal movements is crucial for predicting its impact on flying biodiversity. Most studies on the relationship between wind and seabird movements have, however, focused on local scales, exploring birds perceptive sensitivity to local wind. In this study, we examine low-level wind pattern oscillations in the Southern Indian Ocean at multiple time scales to explain the local- to large-scale movements of the Amsterdam albatross. Adult individuals exhibited smooth trajectories, strongly correlated with seasonal, intra-seasonal or interannual wind oscillations. Conversely, younger individuals displayed more erratic and exploratory movements, often being swept away by eastward moving low-pressure systems at a synoptic time scale. Our results suggest that Amsterdam albatrosses can learn and adapt to the annual and monthly low-level wind climatology and interannual variability of the Southern Indian Ocean. This also highlights the importance of investigating seabird movements in relation to broader-scale wind patterns to support their conservation in a changing climate due to human activities. A robust assessment of regional circulation response to climate change for upcoming decades could help project the impact of climate change on seabird movements and mitigate its effects.

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Socotra Cormorants in the Arabian Gulf represent a large, but isolated population with low genetic diversity

Almansoori, N. M.; Razali, H.; Muzaffar, S. B.; Chabanne, D. B. H.; Natoli, A.; Almusallami, M.; Naser, H.; Khamis, A.; Al Harthi, F.; Aldhaheri, L. S. R.; Alaleeli, M. M. B.; Al Diwani, F. M.; Manlik, O.

2026-04-03 ecology 10.64898/2026.04.01.712451 medRxiv
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The Socotra Cormorant (Phalacrocorax nigrogularis) is a threatened seabird endemic to the coastal areas of the Arabian Gulf and the Arabian Sea, two regions separated by the Strait of Hormuz. Conserving threatened species requires clear delineation of population boundaries and the evaluation of genetic diversity. However, information on population structure and genetic variation, necessary for such an assessment, is lacking for the Socotra Cormorants. In this study, we assessed population structure and genetic diversity of Socotra Cormorants using two contrasting genetic markers: (1) maternally inherited mtDNA cytochrome oxidase 1 (COI) and (2) a nuclear non-coding region, {beta}-fibrinogen intron 7 (FIB7). A total of 279 individuals were sampled from four colonies in the Arabian Gulf and one colony on Hasikiyah Island in the Arabian Sea. Findings based on COI-variation suggest that the Arabian Gulf colonies represent one large population with extensive gene flow between Gulf colonies--except for the most distant pair of colonies--but isolated from Hasikiyah in the Arabian Sea. COI-variation indicated significant differentiation between the colonies inside the Gulf and the Hasikiyah colony. This is consistent with the reported distribution patterns, and may reflect phylogeographic processes of the region. The Gulf population showed substantially lower COI-diversity, with significantly lower nucleotide and haplotype diversity compared to Hasikiyah. In contrast, FIB7 results indicated extensive connectivity among colonies, with no detectable population structure or significant differences between the Gulf population and Hasikiyah. This study presents the first characterization of population structure and genetic diversity of Socotra Cormorants. The low genetic diversity coupled with relative isolation of the Gulf Socotra Cormorants raises conservation concerns regarding their long-term viability by potentially reducing fitness and eroding their evolutionary capacity to adapt to environmental change. LAY SUMMARYO_LIThe Socotra Cormorant is a threatened seabird found in the Arabian Gulf and Arabian Sea, but little was previously known about its population structure and genetic diversity. C_LIO_LIWe analyzed 279 birds from five nesting colonies (4 in the Gulf and 1 in the Arabian Sea), using two genetic markers to assess population connectivity and variation. C_LIO_LIWe found that the Socotra cormorants inside the Gulf appear to form a large, genetically isolated population with relatively low genetic diversity. C_LIO_LIThis is the first study that evaluates population structure and genetic diversity of this endangered seabird. C_LIO_LIThis is important information for the conservation of the Gulf Socotra cormorants because low genetic diversity, coupled with relative isolation, is associated with reduced fitness, and suggests that they may have a lower chance to adapt to environmental changes. C_LI

14
Temporal consistency of judgement biases in bumblebees

Baciadonna, L.; Nityananda, V.

2026-04-09 animal behavior and cognition 10.64898/2026.04.07.716885 medRxiv
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Judgment bias tasks are increasingly used to assess affective states in animals, yet the extent to which they might reflect transient states or stable traits remains unclear. Here, we tested bumblebees (Bombus terrestris) in an active choice task across three repeated sessions to assess individual consistency in the absence of any manipulation. Bees were trained to associate each of two colours with either a high or a low reward, presented in separate chambers. During testing, they were presented with ambiguous colours. Bees were more likely to choose the high reward chamber and to choose more quickly in response to colours closer to the positive colour. The latency to choose the cues showed significant and moderate repeatability across sessions, suggesting a stable, trait-like underlying component. In contrast, the repeatability of the chamber choices was negligible, indicating that such responses might be largely state-dependent and influenced by situational factors. These findings suggest that judgment biases, particularly as assessed through an active choice task reflect states affected by external factors. Active choice tasks may help disentangle stable behavioural traits from transient affective states in invertebrates.

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A versatile cryopreservation method for peri-gastrulation squamate embryos optimised using the veiled chameleon (C. calyptratus)

Weberling, A.; Durnin, M.; Shylo, N. A.; McKinney, M. C.; Wilson, H.; Kupronis, R.; Williams, S. A.; Trainor, P.

2026-04-02 developmental biology 10.64898/2026.04.01.715795 medRxiv
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Stem cell technologies have become a vital component of conservation efforts around the globe. Biobanks and pluripotent stem cell lines help to ensure species and their genetic diversity are preserved. These efforts have however, focussed mostly on mammals and birds, and the cryopreservation protocols for embryos and cells were developed decades ago laying the basis for artificial reproductive techniques for species conservation. With over 20% of non-avian reptile species facing extinction, it is imperative to establish protocols for reptiles to ensure species preservation and also to facilitate the establishment of new reptile model organisms to match the standard of mammals. Here, we have generated a cryopreservation method for preserving early gastrulating veiled chameleon embryos as a representative squamate species. To this end, we first developed a tissue culture method for maintaining cells extracted from peri-gastrulation chameleon embryos and then tested different cryopreservation methods altering the concentration of the penetrating cryoprotectant DMSO and assessing the effect of the addition of non-penetrating cryoprotectants Trehalose and Sucrose. We then optimised a protocol for whole embryo vitrification in 20% DMSO with added Trehalose or Sucrose that can easily be adapted for fieldwork. Taken together, our method not only provides a protocol for conservation efforts but also lays the basis for mechanistic studies of early squamate embryo development by enabling cryopreservation of whole embryos in a fieldwork setting, which facilitates their live transport back to a laboratory for functional experiments or molecular analyses.

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Radiographic assessment of bone maturation as a tool for age estimation in common dolphins (Delphinus delphis)

Hanninger, E.-M. F. F.; Barratclough, A.; Betty, E. L.; Anderson, M. J.; Perrott, M. R.; Bowler, J.; Palmer, E. I.; Peters, K. J.; Stockin, K. A.

2026-04-07 zoology 10.64898/2026.04.05.716530 medRxiv
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We present the first radiographic ageing framework for common dolphins (Delphinus delphis), based on ossification and epiphyseal fusion patterns in the pectoral flipper, demonstrating higher reliability for chronological age estimation than currently available epigenetic approaches for this species. Using individuals of known dental age, we calibrated two modelling approaches to predict dental age from radiographic bone scores: 1) a univariate polynomial regression using a total bone score (sum of 16 scores across all assessed flipper bones), and 2) a multivariate canonical analysis of principal coordinates (CAP) incorporating 16 individual bone-score variables. Both approaches successfully predicted dental age from skeletal ossification patterns. For an age range of 0 to 24 years, polynomial regression demonstrated high predictive accuracy with median absolute errors (MAEs) of 1.25 years in females (Spearmans {rho} = 0.93, R{superscript 2} = 0.90) and 1.08 years in males ({rho} = 0.95, R{superscript 2} = 0.86). The CAP model yielded MAEs of 1.35 years in females ({rho} = 0.90, R{superscript 2} = 0.85) and 1.80 years in males ({rho} = 0.94, R{superscript 2} = 0.84). Notably, both radiographic bone ageing models achieved equal or lower median absolute errors and higher coefficients of determination than a recently developed epigenetic clock for common dolphins derived from the same population (MAE = 1.80, Pearsons correlation (r) = 0.91, R{superscript 2} = 0.82). When applying the bone ageing models to individuals of unknown dental age, both models produced age estimates consistent with expected life-history stages (foetus, neonate, juvenile, subadult, adult), although accuracy declined in dolphins above 20 years, likely as a consequence of subtle age-related variation in skeletal changes in this species. Radiographic ageing provides an accurate non-invasive tool for demographic assessment to support conservation management of common dolphins.

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Physiological consequences of acute heat exposure in mid-gestation on placental, foetal and maternal blood flow using a mouse model.

Francis, S.; Murdoch, C. E.

2026-04-07 physiology 10.64898/2026.04.06.713526 medRxiv
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Pregnant women are particularly susceptible to adverse outcomes from environmental heat, yet the physiological effects of acute heat exposure during pregnancy remain poorly understood. Some physiological changes are monitored in humans; however, investigation of underlying molecular mechanisms requires invasive methods that can only be ethically applied in mammalian models. Moreover, research with animal models has largely focused on early and lethal teratogenic effects of heat exposure and lacks longitudinal physiological monitoring, detailed parameterisation of heating regimes and in-depth investigation of underlying mechanisms. Here we used a mouse model to investigate the impact of a controlled acute heat exposure at mid-gestation (E12{middle dot}5), slowly elevating core body temperature (CBT) over 210mins to raise CBT by [~]1{degrees}C. Using high-frequency ultrasound and morphological analyses, we observed delayed alterations in placental and foetal cerebral blood flow indicative of a brain-sparing response, alongside reduced placental labyrinth zone size. Additionally, maternal cardiac function was impaired, accompanied by cardiac and renal fibrosis and elevated circulating soluble Flt-1 levels, an anti-angiogenic biomarker of gestational hypertension. These findings demonstrate that brief heat stress at mid-gestation can induce lasting effects on placental function and maternal cardiovascular health in a mammalian model, highlighting potential risks for pregnancy outcomes under increasing global temperatures. Together this data suggests that an acute exposure to heat elevating core body temperature by 1{middle dot}2{degrees}C can induce a long-term impact on both placenta and maternal health in a mouse model. It will be important to understand the molecular changes which underpin the pathophysiology and whether this is translated to humans.

18
Insecticide temephos alters thermal dependence of dengue vector

Heffernan, P. M.; Murdock, C. C.; Rohr, J. R.

2026-04-03 ecology 10.64898/2026.04.01.715840 medRxiv
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O_LIAlthough ecological research has long focused on the effects of temperature on population growth, arthropod pests are exposed to a wide variety of environmental factors that affect their performance, such as chemical pesticides targeted against them. Moreover, these environmental factors likely do not act in isolation. Identifying the extent to which abiotic factors interact to affect pest population dynamics can strengthen current and future pest management programs. C_LIO_LIHere, we investigated the extent to which temephos, a common pesticide applied to aquatic environments for mosquito control, influences the thermal performance of juvenile survival and development rate, as well as the intrinsic population growth rate, of the invasive mosquito pest, Aedes aegypti. We implemented a response surface experimental design to measure these traits across seven temperatures and five temephos concentrations and fit temperature- and insecticide-dependent performance curves to assess impacts on the overall performance and the thermal optimum, minimum, and maximum. C_LIO_LITemephos exposure profoundly altered the thermal performance of juvenile survival by reducing survival across all temperatures, shrinking the thermal breadth, and shifting the thermal optimum to warmer temperatures. Through this, temephos also altered the thermal performance of population growth primarily by reducing its thermal breadth. C_LIO_LISynthesis and applications: Our findings demonstrate that interactions between temperature and insecticide exposure can fundamentally reshape pest population dynamics, rather than acting as independent stressors. By quantifying this interaction, we showed that temphos is most effective below the pests thermal optimum, suggesting that larvicides may yield the greatest population suppression in cooler regions or during cooler periods of the year. Incorporating such temperature-dependent efficacy into pest management strategies could improve the timing and spatial targeting of control efforts. More broadly, these results highlight the need to integrate anthropogenic stressors with climatic drivers when predicting pest risk and optimizing management under ongoing environmental change. C_LI

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Semen collection, short term storage, and cryopreservation in the Texas horned lizard (Phrynosoma cornutum)

Julien, A. R.; Griffioen, J. A.; Perry, S. M.; Doege, R.; Burger, I. J.; Barber, D. R.

2026-04-06 zoology 10.64898/2026.04.03.716302 medRxiv
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As global reptile populations continue to decline, improving reproductive success in managed populations of listed species, such as Phrynosoma cornutum (the Texas horned lizard) has become increasingly critical for species survival. One understudied area of reproductive research in reptile species is gamete collection and storage, a crucial component for maintaining genetic diversity. In Texas, semen was collected from wild P. cornutum (n = 20) in June 2025. Semen collection was performed via electroejaculation (EEJ) under alfaxalone anesthesia. Prior to semen collection, snout-vent-lengths (SVL) and weights were recorded and testes measurements were taken using a portable ultrasound. Average sperm motility and concentration across all lizards was 83.7% and 85.7 x 106 sperm/mL, respectively. While lizards with longer SVLs had higher sperm motility, weight and testis size did not affect sperm parameters. Samples were extended in INRA96 and divided for use in cold-storage longevity or cryopreservation trials. Samples under cold-storage conditions were assessed for motility daily for 10 days. Motility was not significantly reduced until 48 hours post-collection and maintained 19% motility at day 10. For cryopreservation, samples were diluted 1:1 in INRAFreeze cryopreservation media and frozen in liquid nitrogen, then immediately thawed. Average post-thaw sperm motility was 13.9%, with the highest post-thaw motility recorded at 38.2%. This is the first report of semen storage and cryopreservation in Phrynosoma and provides valuable insight into semen storage potential in reptile species.

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Growth, survival, and fitness in the first year of life for Pycnopodia helianthoides under different larval densities

Lopez-Nandam, E. H.; Story, L. T.; Evin, M.; Witherly, J.; Upton, M.; Krol, L.; Dunker, F.; Lev, K.

2026-04-06 developmental biology 10.64898/2026.04.02.716152 medRxiv
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Sea star wasting disease has caused widespread mortality in the kelp forest predator, the sunflower sea star (Pycnopodia helianthoides). Wild populations have declined by up to 99% in parts of their native range along the western North American coast. In response, a multi-institutional conservation breeding and rearing program has been initiated to support future reintroduction efforts for the species. We split a full-sibling cohort across four larval density treatments (1 larva/ml, 2 larvae/ml, 5 larvae/ml, and 15-20 larvae/ml) to assess the effects on larval settlement, juvenile survival, and juvenile fitness at 12 months old. Stars raised in the highest density treatment displayed a lower settlement rate and were significantly smaller than the other density groups at 12 months old, but showed no significant difference in flip time, a measure of fitness. Additionally, measurements of diameter, weight, and arm count across modern and historical juvenile and adult stars indicate that P. helianthoides experience exponential weight gain as they grow in length, with corresponding asymptotic growth in arm count. These findings will inform best practices for the aquarium propagation of P. helianthoides and will contribute to broader efforts aimed at reestablishing populations in the wild.