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Journal of Thermal Biology

Elsevier BV

Preprints posted in the last 90 days, ranked by how well they match Journal of Thermal Biology's content profile, based on 14 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
No adaptive plasticity in the heat tolerance of lizard sperm

Wang, W. W.-Y.; Pethe, B.; Ward, A. P.; Wood, W.; Smith, C. T.; Frick, A. J.; Yates, S. S.; Gunderson, A. R.

2026-07-22 ecology 10.64898/2026.07.21.739836 medRxiv
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O_LIAdaptive phenotypic plasticity can help organisms cope with global warming by increasing their heat tolerance, yet most research has focused on whole-organism traits. Much less is known about thermal plasticity at the gamete level despite increasing evidence that reproduction, and particularly sperm function, is especially sensitive to heat. C_LIO_LIWe tested for plasticity in sperm heat tolerance in two ways: via male thermal acclimation (pre-ejaculate stage) and via sperm cell heat hardening (post-ejaculate stage) in brown anole lizards (Anolis sagrei). We also measured plasticity in several other sperm and ejaculate traits in response to male thermal acclimation. C_LIO_LITo test for effects of male thermal acclimation, adult males were exposed to one of two ecologically realistic, fluctuating temperature regimes that either mimic cool spring conditions or projected future summer conditions for eight weeks. Throughout this period, we repeatedly measured sperm heat tolerance (LT50), baseline motility, and sperm count. We also measured sperm morphology at the end of thermal acclimation. We predicted that males in the warmer treatment would produce more heat tolerant sperm. C_LIO_LITo test for post-ejaculate heat hardening, we compared the heat tolerance of ejaculated sperm that either did or did not experience a high but non-lethal temperature prior to heat tolerance measurement. C_LIO_LIWe found no plasticity in sperm heat tolerance due to either male thermal acclimation or sperm cell heat shock. Long-term thermal acclimation of males did not increase sperm heat tolerance, nor change motility, sperm count, or sperm morphology. We also found no evidence for post-ejaculate heat hardening, as exposing sperm cells to mild sublethal heat shock did not enhance sperm heat tolerance. C_LIO_LIOur results indicate that gametic traits have limited capacity for plastic adjustment to thermal stress, which is broadly consistent with the low levels of thermal plasticity found for whole-organism thermal tolerance across ectotherms. This highlights the vulnerability of reproductive traits to rising temperatures and the importance of evolutionary and behavioral responses to buffer organisms from climate change. C_LI

2
Mosquito fertility is (mostly) resilient to sublethal heat shocks

Warusawithana, A. L.; van Heerwaarden, B.; Hoffmann, A. A.; Ross, P. A.

2026-08-05 zoology 10.64898/2026.08.04.742888 medRxiv
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Insects can experience a loss of fertility under acute heat stress, but impacts vary among species and depend on life stage and sex. We examined the effects of sublethal heat shocks on the fertility of three mosquito species (Aedes aegypti, Aedes notoscriptus and Culex quinquefasciatus) when males or females were exposed at the pupal and adult stages. Reproductive performance was assessed through egg-laying success, fecundity, egg hatchability and viable offspring production. Female heat shock had limited effects on fertility in all three species, though exposure of adult females to 41 {degrees}C for 1 hr reduced viable offspring production in Ae. notoscriptus. Male heat shock reduced egg hatchability in Cx. quinquefasciatus and viable offspring production in Ae. aegypti depending on the life stage exposed. We investigated several other aspects of heat shock exposure on Ae. aegypti fertility. In this species, male fertility partially recovered within 24-48 hours following heat shock exposure, while effects of heat shocks on female fertility occurred regardless of whether heat exposure occurred pre- or post-mating. In addition, maternal heat shock exposure enhanced offspring fecundity and viable offspring production under subsequent sublethal heat stress. Overall, our findings demonstrate that mosquito fertility is largely robust to short-term heat shocks, but there are also complex trait, sex, stage, temporal and species-specific effects, with implications for predicting mosquito population persistence and species persistence under future climate extremes.

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Long-term effects of early-life thermal fluctuations on the cellular stress response and CTmax of zebrafish, Danio rerio

Haghighi, H.; Lindsey, B. W.; Jeffries, K. M.

2026-06-08 physiology 10.64898/2026.06.03.729897 medRxiv
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Temperatures in aquatic ecosystems have been affected by anthropogenic activities such as agricultural and industrial water use, and climate change caused by greenhouse gas emissions. Changes in water temperature directly affect the cellular and organismal physiology of fishes because most fishes are ectotherms. These effects can take different forms, such as increased cellular stress and reactive oxygen species (ROS) production. However, the type and magnitude of response depend on the duration and the frequency of the exposure to elevated water temperature. In this study, we investigated the long-term effects of exposure to daily thermal fluctuations occurring during early-life stages of zebrafish, Danio rerio, on gene expression and CTmax in later developmental stages. To do so, wild-type zebrafish were exposed daily to a + 5{degrees}C fluctuation in temperature from ambient (28 {degrees}C) to 33 {degrees}C over the first 30 days post fertilization (dpf), before being held until 90 dpf at ambient temperature. The fish that experienced daily thermal fluctuation were compared to a control group that was kept at 28 {degrees}C throughout the experiment and sampled at the same timepoints. Samples were collected at 18 (larval), 30, 60 (juvenile), and 90 (adult) dpf to study the expression of heat shock proteins and oxidative stress genes. The thermotolerance of fish was tested using CTmax trials at 60 and 90 dpf. Daily thermal fluctuation over the first 30 dpf led to a significant increase in the expression of hsp47, gstp1a, sod1, and sod2 genes at 60 dpf, and hsp47, hsp90aa1, hsp90ab1, cat, glulb, gstp1a, sod1, and sod2 genes at 90 dpf. The only significant increase detected during the larval stage was glulb at 18 dpf. Fish that experienced thermal fluctuation also had a higher CTmax at 60 dpf, but this increased thermotolerance significantly decreased from 60 to 90 dpf, where it was not different between treatments. Overall, our study demonstrates that early-life thermal stress increased cellular stress responses and thermotolerance in zebrafish into later ontological stages.

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Overwintering under the ice: seasonal shifts in cold and hypoxia stress physiology of three winter-active pond insects

Burton, L. S.; Keenan, T. A.; Rodela, T. M.; Toxopeus, J.

2026-08-06 zoology 10.64898/2026.08.05.743153 medRxiv
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Winter poses harsh physiological challenges for insects living in temperate ponds due to the combination of low temperatures and reduced oxygen levels (hypoxia). Despite these stressors, water boatmen (Hesperocorixa sp.), backswimmers (Notonecta sp.) and diving beetles (Laccophilus maculosus) remain active year-round in eastern Nova Scotia, including in ice- covered ponds. However, the underlying mechanisms that allow pond insects to survive overwintering have not been widely studied. We hypothesized that cold and hypoxia tolerance would improve from September to March in these insects, with changes in whole-animal and biochemical correlates of tolerance to both stressors. We collected insects from the field and stocked them in outdoor freshwater mesocosms. Every two months, between September 2023 and April 2024, we characterized whole animal responses and biochemical changes. Whole insect cold tolerance assays indicated a trend of improved ability to sustain voluntary muscle control (CTmin) at lower temperatures in winter-collected insects and higher internal fluid freezing temperatures (SCP). There were no changes in whole animal correlates of hypoxia tolerance over time (surface respiration frequency, submersion and surface time). Potential cryoprotectants like proline, myo-inositol and trehalose increased in concentration in multiple insect taxa during winter but were relatively low compared to terrestrial insect studies. As lactate dehydrogenase activity did not change, there is little evidence that the insects experienced functional hypoxia sufficient to induce anaerobic metabolism. Overall, this research has established a baseline cold and hypoxia tolerance dataset in understudied pond insects. Summary StatementPond insects remain active during low temperature and low oxygen conditions associated with ice cover in winter, supported by physiological and biochemical changes.

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Integrated behavioural, morphological, and reproductive responses reveal a trade-off during diapause in Culex pipiens

Mthawanji, R. R.; Tanianis-Hughes, J.; Binti Rashid, A.; Subramaniam, K. S.; Blagrove, M. S. C.

2026-06-16 ecology 10.64898/2026.06.12.731944 medRxiv
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Diapause is a critical adaptive strategy that enables temperate mosquito species to survive adverse environmental conditions and maintain population persistence across seasons. In Culex pipiens, diapause plays a key role in overwintering and influences the seasonal dynamics of arbovirus transmission. However, diapause expression is often assessed using single traits, limiting our understanding of its integrated physiological basis and variation among populations. In this study, we investigated the behavioural, morphological, and reproductive signatures of diapause across three laboratory strains of Culex pipiens (Mogden, Pirbright, and Pirbright Hybrid) reared under diapause-inducing (10 {degrees}C), cold (14 {degrees}C), and control (26-27 {degrees}C) conditions. We quantified blood-feeding behaviour, wing size as a proxy for somatic growth, and spermatheca size as an indicator of reproductive development. Diapause-inducing conditions resulted in a coordinated phenotype characterised by strong suppression of blood-feeding, increased somatic size, and marked inhibition of reproductive development. Mosquitoes reared at 10 {degrees}C exhibited near-complete feeding inhibition and significantly reduced spermatheca size, consistent with reproductive arrest, while those reared at 14 {degrees}C showed intermediate phenotypes. In contrast, control mosquitoes displayed active feeding and fully developed reproductive structures. Wing size increased progressively with decreasing temperature, with the largest individuals observed under diapause-inducing conditions. When analysed together, wing size and spermatheca development exhibited opposing responses across temperature treatments, revealing a strong negative association and indicating a trade-off between somatic growth and reproductive investment. This integrated response supports the interpretation of diapause as a coordinated life-history strategy involving resource reallocation towards survival. Additionally, diapause expression varied among strains, with the Mogden strain showing reduced sensitivity compared with Pirbright and hybrid populations, highlighting the role of genetic background in diapause plasticity. These findings demonstrate that diapause in Culex pipiens is a multi-trait, plastic phenotype with important implications for overwintering success and the seasonal dynamics of arbovirus transmission in temperate regions.

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Aquatic respiratory rates in red devil vampire crabs (Geosesarma hagen) are dependent on interactions between temperature, sex, and body size

Buck, G.; Juarez, B.; Lacey, M.; O'Connell, L. A.; Watson-Zink, V. M.

2026-07-05 physiology 10.64898/2026.06.30.735571 medRxiv
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The shift to terrestrial environments in ancestrally aquatic animals is often associated with key physiological and physical changes, including shifts in respiratory physiology and in some cases, even the evolution of completely novel respiratory structures. Examining how respiration operates across a gradient of submersion states in ancestrally aquatic terrestrial animals may shed light on how complex biological traits shift under different selective regimes. In this work, we begin exploring respiration in terrestrially-adapted land crabs that still use their gills to respire while underwater. We tested the relationship between aquatic respiratory rates, body size, and sex in red devil vampire crabs (Geosesarma hagen) at two ecologically-relevant temperatures. We found small females respire more than small males at 28{degrees}C, while large females respire more than large males at 21{degrees}C. Additionally, body size is a significant factor affecting respiratory rates of both sexes at 21{degrees}C and warmer temperatures significantly increase respiration in small crabs of both sexes. Interactions between these factors also led to emerging trends that can be explained by both physiological rules, such as reproductive investment and surface-to-volume ratios and heat transfer. We also report a temperature coefficient (Q10) of 1.52 for this species, showing an expected 52% change in respiratory and metabolic rate for every 10{degrees}C increase. This work also demonstrates the importance of understanding how and to what extent biological variables like sex and body size interact with abiotic environmental factors when measuring physiological traits in ectothermic invertebrate animals.

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Sex- and age-specific body condition decline under warmer and drier breeding conditions in European robins

Lopez-Zuluaga, M.; Remacha, C.; Bermejo-Bermejo, A.; Escudero, E.; de la Puente, J.; Perez-Tris, J.

2026-06-10 ecology 10.64898/2026.06.07.725434 medRxiv
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O_LILocal environmental conditions during the breeding season can limit bird populations. Identifying which variables, when, and how they affect key biological traits such as body condition is crucial for understanding long-term population trends under ongoing climate change. C_LIO_LIWe analysed the relationships between environmental variables and body condition during the breeding season in European robins (Erithacus rubecula), aiming to uncover links between short-term environmental influences and long-term trends in body condition in the context of local climate change. C_LIO_LIUsing data from a robin population monitored between 2007 and 2021, we applied weather sliding-window analyses to identify periods when temperature, soil moisture, and vegetation productivity best predicted individual body condition. For each variable, we identified critical time windows (CTWs) influencing (1) body condition across the season and (2) individual changes within two weeks. Juveniles and adults were analysed separately, with adult males and females distinguished during pre- and post-fledging periods. We also assessed long-term trends in environmental variables and body condition, and examined how body condition was correlated with apparent survival. C_LIO_LIBody condition variation across the season was explained by different environmental variables depending on age, sex, and period. Body condition declined with increasing minimum temperatures in adult males and juveniles, and with low soil moisture in adults of both sexes. We did not identify reliable CTWs explaining short-term within-individual changes in body condition. Across 2007-2021, body condition in adult males during the post-fledging period declined with rising minimum temperatures, while fledging dates advanced. Apparent survival was positively associated with body condition only in juvenile robins. C_LIO_LIOur results reveal multiple seasonal environmental influences that may contribute to short- and long-term declines in body condition in European robins, with effects particularly strong (or most detectable) in adult males. Reduced body condition may have demographic consequences by lowering juvenile survival, although shifts in breeding phenology could mitigate this impact. Overall, these findings highlight how environmental effects on body condition can shape long-term population trends and species vulnerability to climate change. C_LI

8
Thermal fertility limits reveal underestimated climatic suitability for Aedes aegypti in temperate regions

Haziqah-Rashid, A.; Metelmann, S.; Stobierska, K.; Gawne, K.; Yu, H.; Sherlock, K.; Baylis, M.; Chrostek, E.; Blagrove, M.

2026-08-07 ecology 10.64898/2026.08.06.743224 medRxiv
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Predicting species distributions under climate change typically relies on thermal limits for survival. However, recent evidence suggests that sublethal temperatures that reduce fertility can constrain distributions more strongly than temperatures causing mortality alone. Despite the importance of mosquitoes as vectors of human disease, thermal fertility limits remain largely uncharacterized in these insects. Here, we show that fertility in Aedes aegypti is highly sensitive to thermal stress and exhibits distinct sex-specific responses. Adult males were more vulnerable to both heat and cold exposure than females. At 38{degrees}C, survival remained high in both sexes despite significant reproductive impairment, indicating that sterility can occur independently of mortality. Similarly, exposure to 2{degrees}C induced male sterility whereas females maintained reproductive function. Morphological analyses of reproductive organs showed temperature-associated damage consistent with the observed fertility loss. Incorporating these fertility thresholds into ecological niche models generated different predictions of climatic suitability compared with models based solely on lethal temperature limits. Our findings demonstrate that sublethal effects on reproduction can substantially influence estimates of mosquito climatic suitability and highlight the importance of incorporating fertility based thermal limits into projections of vector distributions and disease risk under future climate change.

9
Warming limits energetic investment in feeding and post-feeding metabolism in bumblebees

Rossi, N.; Nicholls, E.

2026-06-17 physiology 10.64898/2026.06.11.731625 medRxiv
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Environmental warming is generally expected to increase metabolic demand in ectotherms. However, facultatively endothermic insects such as bumblebees regulate body temperature and may reduce thermogenic investment under warm conditions, potentially altering physiological performance and responses to climate change. We combined flow-through respirometry and infrared thermography to test how elevated ambient temperature (25 vs 35{degrees}C) affects feeding energetics and postprandial metabolism in the bumblebee Bombus terrestris. Bees maintained substantially lower thoracic temperature excess at 35{degrees}C than at 25{degrees}C, both before and during feeding. Feeding metabolic rate was also lower at 35{degrees}C and was strongly positively associated with thoracic temperature excess, indicating that feeding energetics were primarily explained by thermoregulatory state rather than ambient temperature alone. Elevated temperature reduced both the probability and energetic magnitude of specific dynamic action (SDA), including total SDA expenditure, early postprandial metabolism, and peak metabolic amplitude. In contrast, SDA duration and time to peak response showed little temperature dependence. Our results demonstrate that warming can suppress energetic expenditure in facultatively endothermic pollinators by limiting thermogenic investment and postprandial metabolic responses, potentially constraining the energetic flexibility underpinning foraging performance under climate warming.

10
Glucose and methylglyoxal alter plasma protein dynamics, immune traits and glucose homeostasis in a sex- and season-dependent manner in zebra finches

Moreno Borrallo, A.; Criscuolo, F.; Bertile, F.

2026-07-20 physiology 10.64898/2026.07.14.737735 medRxiv
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Birds maintain unusually high circulating glucose levels compared with other vertebrates without developing the diabetic complications observed in mammals, yet the mechanisms underlying this resistance remain unclear. We investigated the effects of chronic glucose and methylglyoxal supplementation on physiological condition in zebra finches (Taeniopygia guttata), with particular emphasis on sex- and season-dependent variation in plasma biochemistry, haematology and immune traits. Ninety zebra finches (45 males, 45 females) were randomly assigned to control, glucose-supplemented (50 g/L), or methylglyoxal (8.33 g/L) drinking treatments. Over one year, we analysed plasma proteins, metabolites (glucose, uric acid, bile acids), tissue damage markers (AST, CK), electrolytes, and immune parameters (leukocyte profiles). Both supplementations increased plasma glucose concentrations, with methylglyoxal producing the strongest effect. More importantly, both treatments disrupted seasonal plasma protein dynamics, preventing the increase in total proteins and globulins normally observed in females during the reproductive period, which suggests alterations in reproductive-related protein metabolism. Glucose supplementation elevated the heterophil-to-lymphocyte (H/L) ratio in May and August, consistent with elevated physiological stress. In contrast, methylglyoxal supplementation reduced the H/L ratio in November and unexpectedly lowered plasma AST and CK concentrations in May, suggesting context-dependent protective effects on tissue integrity despite its well-established pro-oxidative properties, potentially through hormetic mechanisms. Supplementation also modified the calcium/phosphate balance, further supporting treatment effects on seasonal (reproductive) physiology. Overall, our findings demonstrate that glucose and methylglyoxal reshape physiological regulation in zebra finches in a strongly sex- and season-dependent manner rather than simply inducing generalized metabolic damage. These results provide new insights into avian resistance to glucose-associated physiological challenges and highlight the importance of considering both biological context and standardized haematological reference values when investigating glucose metabolism in birds.

11
Freeze tolerance of a beneficial lady beetle, Hippodamia convergens

Ehler, H. E.; Keenan, T. A.; Evans, L. E.; Weisshaar, M. M. R.; Barrett, E. L.; Kirkham, L. J.; Macfarlane, J. K.; Silver, A. B.; Romero, M. F. A.; Alford, B. R.; Rosero, A. M. A.; Clancy, R. P.; Fraser, S. D.; Glennie, G. M.; Hooper, K. E. D.; MacGrath, K. E.; Nauss, J. M.; Pictou, L. A.; Putnam, M. K.; Sturmy, Z. M.; Perry, J. C.; Toxopeus, J.

2026-08-18 zoology 10.64898/2026.08.13.744689 medRxiv
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The convergent lady beetle Hippodamia convergens is widespread in the Americas and considered an important beneficial insect due to its use in pest control. Early work on this species characterized the beetles as freeze-avoidant (freeze-intolerant), suggesting they survive low winter temperatures by physiologically preventing ice formation to temperatures as low as -15{degrees}C. Here, we show that H. convergens can be freeze-tolerant if ice formation occurs at relatively high temperatures. There was 100% survival following inoculative freezing at -0.5{degrees}C and exposure to -3{degrees}C for 20 hours, as well as freezing that spontaneously occurred in fed beetles exposed to -4{degrees}C for 4 hours. Males exposed to 0{degrees}C or -4{degrees}C for 4 hours had similar mating behaviours (latency to first mating, copulation duration) as control beetles exposed to 4{degrees}C, although sample sizes were too small to determine whether freezing itself had an effect on these behaviours. Several putative cryoprotectants were detected in fat body tissue of H. convergens: glycerol, proline, trehalose, and myo-inositol - in order of abundance. Exposure to -3{degrees}C for 20 hours, whether frozen or unfrozen, did not statistically affect cryoprotectant accumulation, although there was a trend towards increased glycerol following freezing. This study is the first to describe inoculative freeze tolerance in a lady beetle and establishes a baseline for future studies that examine the mechanisms underlying this freeze tolerance and the effects of freezing on reproductive behaviour. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=69 SRC="FIGDIR/small/744689v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@1c0382org.highwire.dtl.DTLVardef@12a5932org.highwire.dtl.DTLVardef@145f4adorg.highwire.dtl.DTLVardef@1c2cb22_HPS_FORMAT_FIGEXP M_FIG C_FIG O_LIHippodamia convergens can tolerate freezing at high subzero temperatures C_LIO_LIMating behaviour of males is normal after chilling or freezing C_LIO_LICryoprotectant accumulation may support overwintering survival C_LI

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Thermal conditions and habitat size explain elevational range shift of the northern pika

Sakiyama, T.; Garcia Molinos, J.

2026-08-10 ecology 10.64898/2026.08.06.743106 medRxiv
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AimSpecies in mountain ecosystems often experience upslope distribution shifts in response to climate change. However, elevational range dynamics exhibit substantial complexity around this general trend, with varying magnitude and direction of shifts observed among studies. We tested whether factors other than temperature, such as habitat size and land use, are also responsible for observed elevational shifts in a cold adapted species. LocationHokkaido, Japan MethodsWe resurveyed 61 historical (1963-2007) occurrence sites covering a wide elevational range (60-2,210 m) within the distribution range of the northern pika (Ochotona hyperborea). We assessed the existence of elevational shifts using quantile regression and the relative importance of temperature, habitat size, and land use on the shift using occupancy analysis. ResultsWe detected presence of the northern pika at 41 sites, suggesting extirpations at 20 sites (32.8%) across a wide elevational range of 60-1,550 m. The concentration of extirpation sites at low to mid elevations resulted in a significant upslope shift of the distribution centroid, although the shift was nonsignificant at lower and upper portions of the range. The occupancy analysis revealed a negative effect of long-term mean of summer maximum temperature and a stronger positive effect of habitat size. ConclusionsThis highlights the susceptibility of the northern pika to heat stress and the importance of larger habitats and thus habitat heterogeneity for persistence of local populations. Given the possibility that the observed shift represents a precursor of elevational contraction, continuous monitoring of the local populations is highly needed to evaluate their long-term viability.

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The king of stress? Exploring the physiological resilience and resistance of adult king penguins to chronic glucocorticoid exposure

Cotton, A.; A.Viblanc, V.; Avril, S.; Abolivier, L.; Raymond, E.; Robin, J.-P.; Bize, P.; Blanchard, P.; Stier, A.

2026-08-04 physiology 10.64898/2026.07.31.742038 medRxiv
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To better understand how animals cope with increasingly variable and challenging environments, there is a need to study how prolonged exposure to elevated glucocorticoid hormones (i.e. one mediator of the stress response) affects their physiology. While glucocorticoid elevation is known to increase oxidative stress and accelerate cellular ageing, there is evidence that king penguins (Aptenodytes patagonicus) can prevent oxidative stress during acute stress exposure, suggesting that species may differ in their sensitivity to glucocorticoids downstream negative effects. As king penguins thrive in a seemingly harsh environment, we hypothesized that they may be able to limit the deleterious effects usually associated with chronic glucocorticoid elevation, either through resistance (i.e. prevention of downstream negative effect) or resilience (i.e. rapid recovery following transient negative effect). To test this hypothesis, we experimentally elevated corticosterone levels in incubating king penguins and quantified treatment effects on a suite of physiological traits at multiple time points across incubation and early chick-rearing, up to ca. 2 months after implantation. Corticosterone-treated individuals showed a prolonged increase in corticosterone and decrease in body condition, confirming our treatment likely mimicked sustained stress exposure. Heterophil-to-lymphocyte ratio was only increased transiently, and there was no clear evidence that treatment influenced oxidative stress or telomere length maintenance. Plasma energy metabolites were mainly affected early after implantation, with rapid recovery over time. Overall, our results suggest that adult king penguins show at least moderate resistance and resilience to chronic corticosterone elevation, especially in preventing cellular integrity loss, though at-sea physiological effects remain to be determined.

14
Heatwaves rescue a mosquito host from parasitism across a large geographic gradient

Farner, J. E.; Riley, I. M.; Singh, A. H.; Mordecai, E. A.

2026-06-29 ecology 10.64898/2026.06.26.734620 medRxiv
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The impacts of increasingly frequent and intense heatwaves on parasitism are an important frontier for understanding disease risk under climate change. These impacts are complex because parasitism arises from multiple interacting host and parasite traits that can vary in thermal sensitivity and among populations adapted to different temperature regimes. Here, we used a lab microcosm experiment to investigate the effects of heatwaves occurring during two different phases of a winter-adapted mosquito host - ciliate parasite interaction, for six pairs of sympatric host and parasite populations sourced from two geographic regions with differing histories of winter heat. We found that because heatwaves allowed mosquito larvae to evade infection, they reduced parasitism and increased survival. An early heatwave during initial parasite attack had stronger effects than a later heatwave occurring after infections had established. We did not find evidence of local adaptation to heatwaves: impacts were consistent regardless of population, and were mechanistically predictable from previously measured thermal performance curves that described lower infection and stronger host defenses at warm constant temperatures. The results suggest that increasingly frequent heatwaves may accelerate geographic shifts in parasitism, and demonstrate how fundamental host - parasite thermal biology links to the impacts of extreme temperature events.

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Divergence in thermal performance contributes to ecotype maintenance in an intertidal snail: evidence from in-situ transplants

Dwane, C.; Lorenzo, R.; Galindo, J.; Rolan Alvarez, E.; Truebano, M.

2026-07-24 evolutionary biology 10.64898/2026.07.21.739872 medRxiv
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Physiological adaptation across environmental gradients can contribute to ecological speciation by limiting performance outside locally optimal habitats. Intertidal systems provide strong natural thermal gradients, yet the extent to which thermal physiology contributes to divergence across shore height remains poorly resolved. We investigated cardiac thermal performance in two ecotypes of the marine snail Littorina saxatilis occupying different shore heights along the Galician coast (NW Spain): a wave-adapted ecotype on the lower and mid- shore and a crab-resistant ecotype on the mid- and upper shore. Using infrared photoplethysmography, we quantified heart rate responses in both a reciprocal field transplant experiment and laboratory thermal ramping trials. In the field, the Wave ecotype exhibited significantly higher heart rates than Crab ecotype snails under native mid-shore conditions and after 1 day of exposure to upper-shore conditions. However, after 4 days of exposure to the upper shore, Wave ecotype snails showed a marked reduction in cardiac activity, whereas Crab ecotype populations maintained stable heart rates across transplant locations and durations. In laboratory ramping experiments, Crab ecotypes displayed lower baseline cardiac activity and greater thermal insensitivity across the rising phase of the thermal response curve, while the Wave ecotype exhibited higher cardiac performance and an earlier decline in heartrate at high temperatures. Together, these results demonstrate pronounced ecotype divergence in cardiac thermal physiology and suggest that chronic exposure to upper-shore conditions compromises cardiac performance in the Wave ecotype. Such physiological differences likely contribute to vertical zonation and the evolution of barriers to gene flow between these ecotypes.

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The impact of temperature-induced vertebral anomalies on C-start swimming performance in Astyanax mexicanus (Teleostei: Characidae)

Arnold, K. M.; Reyes-Corral, W. D.; Howard, O.; Graca, C.; Aguirre, W. E.

2026-08-11 zoology 10.64898/2026.08.06.743195 medRxiv
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This study investigates the impact temperature-induced vertebral anomalies have on the C-start escape response of Astyanax mexicanus, a model species in evolutionary developmental biology. Employing three temperature treatments to induce varying degrees of skeletal anomalies, we assessed their effects on key swimming performance metrics including, C-start time, curvature coefficient, head displacement distance, and displacement velocity. Through the use of linear mixed models and generalized linear mixed models, our results reveal that specific anomalies such as vertebral fusions and anomalous haemal and neural spines affected the curving ability of C-start escape responses. However, these did not negatively impact other performance parameters, with velocity, distance, and response time showing no significant impacts from any anomaly types, when assessed individually. This suggests a complex interplay between structural deformities and compensatory physiological mechanisms that maintain functional performance. Other variables measured had a stronger and significant impact on swimming performance, including standard length, vertebral number, and temperature treatment, which influenced escape speed, curving ability, and overall locomotor performance. Our findings challenge conventional perceptions about the debilitating impact of vertebral anomalies, indicating that many affected fish can still effectively perform escape maneuvers critical for survival.

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Continuous thermal sensitivity of gene expression following acclimation in Drosophila subobscura

Tushar, E.; Heilig, M.; Haddad, A.; DeMayo, J. A.; Ragland, G.

2026-08-11 evolutionary biology 10.64898/2026.08.05.743044 medRxiv
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The physiology of ectotherms can change substantially during acclimation to changing environmental temperature. The role of transcription in acclimation responses has been well-established, but it remains unclear whether transcriptional regulation generally reflects abrupt changes after surpassing temperature thresholds, or whether transcript abundance is a relatively monotonic, continuous function of acclimation temperature. In this study we exposed adult male Drosophila subobscura flies to four different 96-hour acclimation treatments at temperatures that were not acutely stressful but ranged from relatively cold (10{degrees}C) to relatively warm (27{degrees}C) with respect to standard rearing conditions. Transcriptome sequencing of whole-body homogenates (mRNAseq) revealed a massive, transcriptome-wide response across acclimation temperatures, with a marked overrepresentation of genes that were continuously and monotonically up- and down-regulated in response to increasing acclimation temperature. Though some genes showed more complex relationships consistent with putative threshold responses, a high percentage of the differentially expressed transcriptome (42%) showed continuous and strictly monotonic relationships. Functional enrichment suggested continuous up-regulation of spermatogenesis-related transcripts with increasing temperature and continuous up-regulation of oxidative phosphorylation-related transcripts with decreasing temperature, illustrating contrasting patterns consistent with previous studies of thermal sensitivity of male reproduction and metabolic compensation in the cold. Thus, continuous thermal sensitivity of transcription is a hallmark of acclimation in D. subobscura that likely underlies the continuous thermal sensitivity of downstream physiological processes. We also provide evidence for shared transcriptomic responses across short-term acclimation (this study) vs. published results for long-term, developmental acclimation.

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Maternal age at reproduction drives multigenerational variation in life-history traits

Zhang, K.; SCHAUSBERGER, P.; Holwell, G.; Zhang, Z.-Q.

2026-07-29 zoology 10.64898/2026.07.28.741326 medRxiv
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Offspring phenotypes are often influenced by maternal age at reproduction. However, the persistence of such effects across multiple generations remains poorly understood, especially in rare cases where offspring of older mothers exhibit enhanced performance. Using the predatory mite Amblyseius herbicolus, we experimentally tested whether maternal age at reproduction influences offspring life history across two generations under contrasting dietary conditions. We found that grandmaternal (F0) and maternal (F1) age affected offspring (F2) developmental time and body size, but not hatching success, survival, or oviposition. The effect of maternal age on body size depended on grandmaternal age, suggesting that grandmaternal age mediates how mothers adjust offspring provisioning. Diet strongly influenced offspring traits but showed limited interaction with age effects. To our knowledge, this is the first study to demonstrate the persistence of an inverse Lansing effect across generations, whereby enhanced immature development is maintained without detectable trade-offs in reproduction or adult survival. These results identify maternal age as a key driver of transgenerational phenotypic variation, with important implications for population dynamics and responses to environmental change at the population level.

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Can urban fence lizards identify cats?

Pfister, E. M.; Blumstein, D. T.; Shaffer, H. B.; Daversa, D. R.

2026-07-23 ecology 10.64898/2026.07.22.735083 medRxiv
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Urban habitats can serve as wildlife refuges, or as ecological traps. As global urbanization continues to increase, native prey species may experience reduced threat from natural predators that tend to avoid human-populated areas. However, such protection often comes at the expense of susceptibility to synanthropic predators, one of the most prevalent being domestic house cats (Felis catus). While cats are a known driving force of mortalities in urban wildlife, their sublethal effects remain understudied. To better understand the behavioral effects of synanthropic predators on native urban prey, we examined responses of western fence lizards (Sceloporus occidentalis) to Artificial Intelligence (AI)-generated visual stimuli of native predators (snakes) and synanthropic predators (house cats). Visual stimuli included: California striped racers (Masticophis lateralis), a native predator; domestic cats, a synanthropic mammalian predator (Felis catus); and desert cottontails (Sylvilagus audubonii), a native mammalian non-predator. We quantified lizard responses as the change in rates of looking and locomotion during and following the video presentation. Our results revealed that lizards discriminated cats from the control by looking more within the first 90 seconds following exposure. Lizards looked more across the entire 5-minute observation period following exposure to all stimulus types compared to control conditions, indicating that AI generated visual stimuli effectively elicit behavioral responses. Together, these findings indicate that urban lizards initially detect and respond to visual cues of cats, suggesting that cat-induced mortality is not a result of inability to detect such predators. Thus, cats influence urban prey not only through direct predation risk, but also changes in behavior associated with the presence of synanthropic predators.

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Contrasting effects of glucose and methylglyoxal supplementation on blood oxidative status, blood cells' telomere dynamics and apoptosis in birds

Moreno Borrallo, A.; Colominas-Ciuro, R.; Colicchio, B.; M'kacher, R.; Allak, A. L.; Criscuolo, F.; Bertile, F.

2026-07-13 physiology 10.64898/2026.07.09.737063 medRxiv
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Birds exhibit longer lifespans than similarly sized mammals, despite having higher mass-adjusted blood glucose levels. This makes them a valuable model for the comparative study of the metabolic and physiological aspects of aging. Circulating glucose contributes to multiple pathological processes, primarily through glycation reactions and the formation of advanced glycation end-products (AGEs), as well as by promoting oxidative stress. These mechanisms are interconnected by feedback loops and play a key role in the development of age-related pathologies. To explore the causal role of glycaemia in avian ageing, we conducted a one-year dietary supplementation experiment in captive zebra finches. Birds received either glucose- or methylglyoxal-enriched water. Previously, we observed that chronic glucose supplementation in zebra finches increased mortality, an effect that did not appear to be mediated by the associated increase in plasma protein glycation or AGE levels. Therefore, the mechanisms underlying increased mortality in the glucose group remained unclear. In the present study, we investigated how glucose and methylglyoxal supplementation affect blood oxidative status and red blood cell telomere dynamics and apoptosis. We found that methylglyoxal supplementation decreased the non-enzymatic antioxidant capacity (OXY) of plasma and increased DNA damage, while glucose supplementation had no significant effect on oxidative stress, although circulating glucose levels influenced oxidative status in a sex-dependent manner. Males exhibited a positive correlation between glucose levels and organic hydroperoxides and protein carbonyls. Additionally, we report, for the first time in birds, a seasonal variation in telomere length, which was more pronounced in glucose-supplemented individuals, yet seemed independent of oxidative status. Apoptosis probability increased with both treatments, particularly with the methylglyoxal supplementation. These results highlight that glucose and methylglyoxal trigger different glucotoxicity-related pathways, with distinct effects on bird health and aging. However, the relationship between glucose supplementation and mortality remains still unclear and warrants further investigation.