COVID-19 as a chronic stressor and the importance of individual identity: A data-driven look at academic productivity during the pandemic.

The COVID-19 pandemic impacted personal and professional life. For academics, research, teaching, and service tasks were upended and we all had to navigate the altered landscape. However, some individuals faced a disproportionate burden, particularly academics with minoritized identities or those who were early career, were caregivers, or had intersecting identities. As comparative endocrinologists, we determine how aspects of individual and species-level variation influence response to, recovery from, and resilience in the face of stressors. Here, we flip that framework and apply an integrative biological lens to the impact of the COVID-19 chronic stressor on our endocrine community. We address how the pandemic altered impact factors of academia (e.g., scholarly products) and relatedly, how factors of impact (e.g., sex, gender, race, career stage, caregiver status, etc.) altered the way in which individuals could respond. We predict the pandemic will have long-term impacts on the population dynamics, composition, and landscape of our academic ecosystem. Impact factors of research, namely journal submissions, were altered by COVID-19, and women authors saw a big dip. We discuss this broadly and then report General and Comparative Endocrinology (GCE) manuscript submission and acceptance status by gender and geographic region from 2019 to 2023. We also summarize how the pandemic impacted individuals with different axes of identity, how academic institutions have responded, compile proposed solutions, and conclude with a discussion on what we can all do to (re)build the academy in an equitable way. At GCE, the first author positions had gender parity, but men outnumbered women at the corresponding author position. Region of manuscript origin mattered for submission and acceptance rates, and women authors from Asia and the Middle East were the most heavily impacted by the pandemic. The number of manuscripts submitted dropped after year 1 of the pandemic and has not yet recovered. Thus, COVID-19 was a chronic stressor for the GCE community.

Transgenerational effects of maternal corticosterone across early life in a viviparous snake.

Glucocorticoids (GCs) are central mediators of vertebrate responses to intrinsic and extrinsic stimuli. Among the sources of variation in circulating GCs are transgenerational effects mediated by mothers. Here we studied potential maternal effects mediated by GCs on offspring phenotype in a live-bearing reptile, the western terrestrial garter snake (Thamnophis elegans). We evaluated the association between baseline corticosterone (CORT) levels during gestation (i.e., preparturition) in field-captured mothers and 1) reproductive success and offspring sex ratios, 2) birth phenotypic traits of offspring born under common-garden laboratory conditions, and 3) neonate (age 3 months) and juvenile (age 12 months) traits of offspring raised under two thermal regimes ('warm' and 'cool') during their first year of life. Reproductive success and offspring sex ratios were not associated with preparturition maternal CORT, but pregnant snakes with higher CORT levels gave birth to smaller, lighter offspring, which tended to grow faster to age three months. Neonate baseline CORT varied with preparturition maternal CORT in a sex-specific manner (positive trend for females, negative for males). Maternal CORT effects on offspring phenotype were no longer detectable in juveniles at age one year. Instead, juvenile phenotypes were most influenced by rearing environment, with offspring raised under the cool regime showing higher baseline CORT and slower growth than those raised under warmer conditions. Our findings support the notion that offspring phenotype might be continuously adjusted in response to environmental cues -both pre- and post-natal- and that the strength of maternal CORT effects declines as offspring develop and experience unique environmental challenges. Our results contribute to a growing literature on transgenerational effects of hormones and help to fill a gap in our knowledge of these effects in ectothermic amniotes.

Anthropogenic impact on the reproductive health of two wild Patagonian fish species with differing reproductive strategies.

A particularly concerning outcome of environmental pollution is the disturbance of reproductive processes. However, studies on the impacts of pollution on the reproductive health of fish inhabiting South American environments are limited. We studied the impact of anthropogenic pollution on the reproductive health of two sympatric Patagonian marine fish species with different reproductive strategies: the live-bearing rockfish Sebastes oculatus and the egg-laying Brazilian sandperch Pinguipes brasilianus. Our findings reveal that both species presented some degree of reproductive disturbance when inhabiting an affected site, but the specific alterations differed depending on the species, sex, and season. During the reproductive season, 17ß-estradiol levels were elevated in females of both species living in polluted areas, while no differences in androgen levels were observed in either species or season. The gonadosomatic index (GSI) was affected in both sexes of S. oculatus during the non-reproductive season, while the gonadal stages were mainly affected in both sexes of P. brasilianus. No signs of intersex condition were observed. Our results highlight the importance of including diverse reproductive parameters to better understand anthropogenic effects on wild animals. Long-term studies including other fish species and including offspring (to evaluate possible transgenerational effects) will be necessary to determine the consequences of the documented reproductive alterations, particularly whether fish species inhabiting Patagonian marine reef areas will be able to reproductively adapt to increasing marine anthropogenic disturbances.

Epitheliocystis in wild marine fishes and its relation with anthropogenic pollution.

Epitheliocystis is a fish disease with recorded in increasing numbers globally. Here we show that (a) pollution-exposed wild marine fishes have higher infection prevalence than nonexposed individuals from reference sites, suggesting that anthropogenic pollution predisposes individuals to this disease in natural systems, (b) the effect of pollution on infection status did not vary significantly between two sympatric fish species, and (c) infected and noninfected individuals from both species did not differ significantly in selected immune and general-health parameters, a result that deserves further study with increased sample sizes.

Genetic background and thermal environment differentially influence the ontogeny of immune components during early life in an ectothermic vertebrate.

An understudied aspect of vertebrate ecoimmunology has been the relative contributions of environmental factors (E), genetic background (G) and their interaction (G × E) in shaping immune development and function. Environmental temperature is known to affect many aspects of immune function and alterations in temperature regimes have been implicated in emergent disease outbreaks, making it a critical environmental factor to study in the context of immune phenotype determinants of wild animals. We assessed the relative influences of environmental temperature, genetic background and their interaction on first-year development of innate and adaptive immune defences of captive-born garter snakes Thamnophis elegans using a reciprocal transplant laboratory experiment. We used a full-factorial design with snakes from two divergent life-history ecotypes, which are known to differ in immune function in their native habitats, raised under conditions mimicking the natural thermal regime-that is, warmer and cooler-of each habitat. Genetic background (ecotype) and thermal regime influenced innate and adaptive immune parameters of snakes, but in an immune-component specific manner. We found some evidence of G × E interactions but no indication of adaptive plasticity with respect to thermal environment. At the individual level, the effects of thermal environment on resource allocation decisions varied between the fast- and the slow-paced life-history ecotypes. Under warmer conditions, which increased food consumption of individuals in both ecotypes, the former invested mostly in growth, whereas the latter invested more evenly between growth and immune development. Overall, immune parameters were highly flexible, but results suggest that other environmental factors are likely more important than temperature per se in driving the ecotype differences in immunity previously documented in the snakes under field conditions. Our results also add to the understanding of investment in immune development and growth during early postnatal life under different thermal environments. Our finding of immune-component specific patterns strongly cautions against oversimplification of the highly complex immune system in ecoimmunological studies. In conjunction, these results deepen our understanding of the degree of immunological flexibility wild animals present, information that is ever more vital in the context of rapid global environmental change.

Ecotourism effects on health and immunity of Magellanic penguins at two reproductive colonies with disparate touristic regimes and population trends.

Negative effects of ecotourism on wildlife are rising worldwide. Conservation physiology can play a major role in protecting wildlife by providing early alerts on changes in the status of individuals exposed to tourist activities. We measured an integrated set of immune and health-state indices to evaluate the effects of ecotourism on Magellanic penguins (Spheniscus magellanicus). We studied two reproductive colonies that differed in the intensity of tourism and population trends: Punta Tombo (higher tourism intensity, declining population) and San Lorenzo (lower tourism intensity, growing population). Within each colony, we compared individuals from an area that was exposed to tourists and a control area where tourism was excluded. Adult penguins exposed to tourism at Punta Tombo, but not at San Lorenzo, showed physiological alterations indicative of chronic stress (higher heterophil to lymphocyte ratios) and parasitic infection (elevated heterophil and eosinophil counts). Penguin chicks exposed to tourism at Punta Tombo, but not at San Lorenzo, also showed physiological alterations indicative of poor immune and general-health condition: lower humoral innate immunity, haematocrit, and glucose levels and higher inflammatory responses likely due to increased prevalence of fleas. Our results indicate that individuals of a declining population exposed to high levels of tourism express physiological indicators of chronic stress and poor health that could make adults and juveniles vulnerable to disease. These effects are expressed despite a long history of exposure and behavioural habituation to human visitation. In contrast, individuals of a growing population exposed to more recent and lower levels of tourism showed no effect. Our study demonstrates how a diverse physiological toolkit within a conservation physiology approach can provide important information for a better comprehension of anthropogenic effects on wild animals in our changing world.

Merging the "Morphology-Performance-Fitness" Paradigm and Life-History Theory in the Eagle Lake Garter Snake Research Project.

SYNOPSIS: The morphology-performance-fitness paradigm for testing selection on morphological traits has seen decades of successful application. At the same time, life-history approaches using matrix methods and perturbation studies have also allowed the direct estimate of selection acting on vital rates and the traits that comprise them. Both methodologies have been successfully applied to the garter snakes of the long-term Eagle Lake research project to reveal selection on morphology, such as color pattern, number of vertebrae, and gape size; and life-history traits such as birth size, growth rates, and juvenile survival. Here we conduct a reciprocal transplant study in a common laboratory environment to study selection on morphology and life-history. To place our results in the ecomorphology paradigm, we measure performance outcomes (feeding rates, growth, insulin-like growth factor 1 titers) of morphological variation (body size, condition) and their fitness consequences for juvenile survival-a trait that has large fitness sensitivities in these garter snake populations, and therefore is thought to be subject to strong selection. To better merge these two complementary theories, we end by discussing our findings in a nexus of morphology-performance-fitness-life history to highlight what these approaches, when combined, can reveal about selection in the wild.

PHYSIOLOGIC PARAMETERS AND THEIR RESPONSE TO HANDLING STRESS IN A NEOTROPICAL MIGRATORY SHOREBIRD DURING THE NONBREEDING SEASON.

Physiologic traits are promising indicators of population health in the face of rapidly changing environments. We obtained values of diverse physiologic parameters for Two-banded Plovers (Charadrius falklandicus) in coastal sites in Patagonia, Argentina, with the objectives of determining the timeline in which these parameters become affected by the stress of capture and handling and of obtaining reference values for future monitoring of these populations. We analyzed packed cell volume, white blood cell profile, heterophil/lymphocyte ratio, bacterial agglutination titer, and total protein, glucose, triglyceride, and cholesterol levels in apparently healthy birds. Glucose, total white blood cell count, lymphocytes, and eosinophil levels showed changes with handling times >60 min after capture. The remaining parameters did not manifest significant alterations in response to capture and handling of up to 232 min (average=105.2, SD=56.7). Therefore, although researchers should attempt to obtain blood samples as soon as possible after capture, inclusion of physiologic parameters in monitoring studies of species not easily sampled in a few minutes, such as Two-banded Plovers and other shorebird species during migration, should not be discouraged. Here we provide a physiologic report for the species that can be considered as reference values during the nonbreeding season at Patagonian coastal sites.

Immune variation during pregnancy suggests immune component-specific costs of reproduction in a viviparous snake with disparate life-history strategies.

Growing evidence suggests the existence of trade-offs between immune function and reproduction in diverse taxa. Among vertebrates, however, there is still a taxonomic bias toward studies in endotherms, particularly birds. We tested the hypothesis that reproduction entails immune-related costs in the viviparous garter snake, Thamnophis elegans, from populations that exhibit two life-history strategies, termed ecotypes, with contrasting paces of life. Between the two ecotypes, we predicted lower immune function in gravid than non-gravid females of both strategies, but with relatively larger immunity costs in the ecotype that generally invests more in current reproduction. Across individuals, we predicted greater immune costs for females investing more in the present specific reproductive event (i.e., higher fecundity) irrespective of their ecotype. We assessed leukocyte profiles and measured bactericidal capacity of plasma (innate immunity) and T- and B-lymphocyte proliferation (adaptive immunity) in gravid and non-gravid females in their natural habitats. We also collected data on reproductive output from these same gravid females brought into captivity. Gravid females of both ecotypes showed lower T-lymphocyte proliferation responses to concanavalin A than non-gravid females, but no differential costs were observed between ecotypes. The remaining immune variables did not vary between gravid and non-gravid females. Among gravid females within each ecotype, those with larger reproductive output showed lower total leukocyte counts, suggesting a fecundity-dependent trade-off. Our study contributes to the comparative ecoimmunology of vertebrates by highlighting the immune component-specificity of trade-offs between reproduction and immune function and showing that costs can be fecundity-dependent in some, but not all cases.

Vertical Transmission of Hepatozoon in the Garter Snake Thamnophis elegans.

Vertical transmission of blood parasites has been demonstrated in humans and some domestic species, but it has not been well documented in wild populations. We assessed whether Hepatozoon blood parasites are vertically transmitted in naturally infected individuals of the viviparous western terrestrial garter snake ( Thamnophis elegans ). Blood smears were taken from nine wild-caught gravid female snakes at capture, preparturition, and postparturition, and then from their laboratory-born offspring at age 2 mo and 1 yr. All infected offspring were born to four infected females, although not all offspring in a given litter were necessarily infected. Parasites were not detected in offspring born to the five uninfected mothers. The highest parasite loads were found in neonates at 2 mo of age. Parasite prevalence did not vary between sexes in offspring, but females showed higher loads than did males when 2 mo old. This study supports vertical transmission of Hepatozoon in naturally infected viviparous snakes and suggests that vertical transmission of hematozoan parasites might be an overlooked mode of transmission in wildlife.

Insulin-like signaling (IIS) responses to temperature, genetic background, and growth variation in garter snakes with divergent life histories.

The insulin/insulin-like signaling pathway (IIS) has been shown to mediate life history trade-offs in mammalian model organisms, but the function of this pathway in wild and non-mammalian organisms is understudied. Populations of western terrestrial garter snakes (Thamnophis elegans) around Eagle Lake, California, have evolved variation in growth and maturation rates, mortality senescence rates, and annual reproductive output that partition into two ecotypes: "fast-living" and "slow-living". Thus, genes associated with the IIS network are good candidates for investigating the mechanisms underlying ecological divergence in this system. We reared neonates from each ecotype for 1.5years under two thermal treatments. We then used qPCR to compare mRNA expression levels in three tissue types (brain, liver, skeletal muscle) for four genes (igf1, igf2, igf1r, igf2r), and we used radioimmunoassay to measure plasma IGF-1 and IGF-2 protein levels. Our results show that, in contrast to most mammalian model systems, igf2 mRNA and protein levels exceed those of igf1 and suggest an important role for igf2 in postnatal growth in reptiles. Thermal rearing treatment and recent growth had greater impacts on IGF levels than genetic background (i.e., ecotype), and the two ecotypes responded similarly. This suggests that observed ecotypic differences in field measures of IGFs may more strongly reflect plastic responses in different environments than evolutionary divergence. Future analyses of additional components of the IIS pathway and sequence divergence between the ecotypes will further illuminate how environmental and genetic factors influence the endocrine system and its role in mediating life history trade-offs.

Effects of a novel climate on stress response and immune function in painted turtles (Chrysemys picta).

Climate change may subject animals to increasingly stressful environmental conditions, which could have negative physiological consequences if stress levels are elevated for long periods. We conducted a manipulative experiment to determine the effects of a novel climate on stress levels and immune function in a model reptile species, the painted turtle. We collected turtles from four populations across the species' geographic range and housed them in a common-garden in one population's local climate. We measured levels of the stress hormone corticosterone and tested two aspects of innate immune function, bactericidal capacity and natural antibody agglutination, at the time of capture (baseline) and three additional time points over 1 year. The four populations did not differ in corticosterone levels over the course of 1 year, and corticosterone levels were also similar at each sampling period except that post-hibernation corticosterone levels were significantly lower than the previous three time points. Furthermore, we found no evidence that elevated corticosterone depressed immune function in the painted turtle. Our study suggests that turtles exposed to novel climatic conditions did not display a detectable stress response, nor did the novel climate depress immune function in the transplanted populations. Therefore, in terms of innate immune function, turtles may be relatively resilient to at least small changes in climatic conditions.

Physiological indices of stress in wild and captive garter snakes: correlations, repeatability, and ecological variation.

Glucocorticoids and leukocyte ratios have become the most widespread variables employed to test hypotheses regarding physiological stress in wild and captive vertebrates. Little is known, however, regarding how these two indices of stress covary in response to stressors, their repeatability within individuals, and differences in response time upon capture. Furthermore, few studies compare stress indices between captive and wild populations, to assess potential alteration of stress physiology in captivity. To address these issues, we examined corticosterone (CORT) and heterophil to lymphocyte (H:L) ratios in two ecotypes of the garter snake Thamnophis elegans. We found that CORT and H:L ratios were not correlated within individuals, and both variables showed little or no repeatability over a period of months. CORT levels, but not H:L ratios, were higher for individuals sampled after 10min from the time of capture. However, both variables showed similar patterns of ecotypic variation, and both increased over time in gravid females maintained in captivity for four months. We suggest that CORT and H:L ratios are both useful, but disparate indices of stress in this species, and may show complex relationships to each other and to ecological and anthropogenic variables.

Complex interplay of body condition, life history, and prevailing environment shapes immune defenses of garter snakes in the wild.

The immunocompetence "pace-of-life" hypothesis proposes that fast-living organisms should invest more in innate immune defenses and less in adaptive defenses compared to slow-living ones. We found some support for this hypothesis in two life-history ecotypes of the snake Thamnophis elegans; fast-living individuals show higher levels of innate immunity compared to slow-living ones. Here, we optimized a lymphocyte proliferation assay to assess the complementary prediction that slow-living snakes should in turn show stronger adaptive defenses. We also assessed the "environmental" hypothesis that predicts that slow-living snakes should show lower levels of immune defenses (both innate and adaptive) given the harsher environment they live in. Proliferation of B- and T-lymphocytes of free-living individuals was on average higher in fast-living than slow-living snakes, opposing the pace-of-life hypothesis and supporting the environmental hypothesis. Bactericidal capacity of plasma, an index of innate immunity, did not differ between fast-living and slow-living snakes in this study, contrasting the previously documented pattern and highlighting the importance of annual environmental conditions as determinants of immune profiles of free-living animals. Our results do not negate a link between life history and immunity, as indicated by ecotype-specific relationships between lymphocyte proliferation and body condition, but suggest more subtle nuances than those currently proposed.

Interrelations among immune defense indexes reflect major components of the immune system in a free-living vertebrate.

Understanding the relationships among immune components in free-living animals is a challenge in ecoimmunology, and it is important not only for selecting the immune assays to be used but also for more knowledgeable interpretation of results. In this study, we investigated the relationships among six immune defense indexes commonly used by ecoimmunologists and measured simultaneously in individual free-living tree swallows. Three main axes of variation in immune function were identified using a principal components analysis, representing variation in T-cell, B-cell, and innate immunity. Measures within each axis tended to be positively correlated among individuals, while measures in different axes were uncorrelated. A trade-off between T-cell function and B-cell function became apparent only when variation among individuals in body condition, age, and general quality was taken into account. Interestingly, the level of natural antibodies, a component of innate immunity, showed the strongest association with components of acquired B-cell function, possibly reflecting a common underlying genetic mechanism, as has been documented in poultry. Our results indicate that despite the complexity of the immune system, important insights can be gained by using the currently available assays but in a more comprehensive approach than has generally been used in the field of ecoimmunology.

Complete penoscrotal transposition: A three-stage procedure.

Complete penoscrotal transposition (CPST) with an intact scrotum is a rare anomaly in which the scrotum is located cephalic to the penis. It is the most severe degree of malformation of a spectrum of abnormalities in scrotal development. There are few cases reported in the literature, and there are few descriptions of the technique for correction and results. We describe a new case of CPST and its sequential correction.

Corticosterone and pace of life in two life-history ecotypes of the garter snake Thamnophis elegans.

Glucocorticoids are main candidates for mediating life-history trade-offs by regulating the balance between current reproduction and survival. It has been proposed that slow-living organisms should show higher stress-induced glucocorticoid levels that favor self-maintenance rather than current reproduction when compared to fast-living organisms. We tested this hypothesis in replicate populations of two ecotypes of the garter snake (Thamnophis elegans) that exhibit slow and fast pace of life strategies. We subjected free-ranging snakes to a capture-restraint protocol and compared the stress-induced corticosterone levels between slow- and fast-living snakes. We also used a five-year dataset to assess whether baseline corticosterone levels followed the same pattern as stress-induced levels in relation to pace of life. In accordance with the hypothesis, slow-living snakes showed higher stress-induced corticosterone levels than fast-living snakes. Baseline corticosterone levels showed a similar pattern with ecotype, although differences depended on the year of study. Overall, however, levels of glucocorticoids are higher in slow-living than fast-living snakes, which should favor self-maintenance and survival at the expense of current reproduction. The results of the present study are the first to relate glucocorticoid levels and pace of life in a reptilian system and contribute to our understanding of the physiological mechanisms involved in life-history evolution.

Consequences of immune system aging in nature: a study of immunosenescence costs in free-living Tree Swallows.

Immunosenescence, the aging of the immune system, is well documented in humans and laboratory models and is known to increase infection risk, morbidity, and mortality among the old. Immunosenescence patterns have recently been unveiled in various free-living populations, but their consequences in the wild have not been explored. We investigated the consequences of immunosenescence in free-living Tree Swallows Tachycineta bicolor through a field experiment simulating a bacterial infection (challenge with lipopolysaccharide, LPS) in females of different ages during the nestling rearing period. We assessed behavioral and physiological responses of females, as well as growth and quality of their offspring, to determine the costs associated with the simulated infection. Results of the experiment differed between the two years of study. In the first year, old females challenged with LPS lost more body mass and reduced their nest visitation rates more, and their offspring tended to grow slower compared to similarly challenged younger females. In contrast, in the second year, old females did not appear to suffer larger costs than younger ones. Interestingly, immunosenescence was only detected during the first year of the study, suggesting that it is the dysregulated immune function characteristic of immunosenescent individuals rather than age per se that can lead to higher costs of immune defense in old individuals. These results provide the first evidence of costs of immunosenescence in free-living animals and support the hypothesis that old, immunosenescent individuals pay higher costs than younger ones when faced with a challenge to their immune system. Our results also suggest that these costs are mediated by an exaggerated sickness behavior, as seen in laboratory models, and can be modulated by ecological factors such as weather conditions and food availability.

Developmental plasticity of immune defence in two life-history ecotypes of the garter snake, Thamnophis elegans - a common-environment experiment.

1. Ecoimmunological theory predicts a link between life-history and immune-defence strategies such that fast-living organisms should rely more on constitutive innate defences compared to slow-living organisms. An untested assumption of this hypothesis is that the variation in immune defence associated with variation in life history has a genetic basis. 2. Replicate populations of two life-history ecotypes of the garter snake Thamnophis elegans provide an ideal system in which to test this assumption. Free-ranging snakes of the fast-living ecotype, which reside in lakeshore habitats, show higher levels of three measures of constitutive innate immunity than those of the slow-living ecotype, which inhabit meadows around the lake. Although this pattern is consistent with the ecoimmunological pace-of-life hypothesis, environmental differences between the lakeshore and meadow habitats could also explain the observed differences in immune defence. 3. We performed a common-environment experiment to distinguish between these alternatives. Snakes born and raised in common-environment conditions reflected the immune phenotype of their native habitats when sampled at 4 months of age (i.e. fast-living lakeshore snakes showed higher levels of natural antibodies, complement activity and bactericidal competence than slow-living meadow snakes), but no longer showed differences when 19 months old. 4. This suggests that the differences in innate immunity observed between the two ecotypes have an important - and likely age-specific - environmental influence, with these immune components showing developmental plasticity. A genetic effect in early life may also be present, but further research is needed to confirm this possibility and therefore provide a more definitive test of the ecoimmunological pace-of-life hypothesis in this system.

Ontogeny of innate and adaptive immune defense components in free-living tree swallows, Tachycineta bicolor.

Little is known about the development of immune function in wild animals. We investigated the ontogeny of immune defense in a free-living bird, the tree swallow. We assessed total and differential leukocyte counts, natural antibodies, complement activity, in vivo skin swelling response, and in vitro lymphocyte proliferation and compared the levels of development between nestlings and young adults. We also assessed whether body condition explained variation in these immune components. We found some support for the prediction that innate defenses, which do not need to generate a broad repertoire of specific receptors, would reach adult levels earlier than adaptive defenses. In contrast, we found limited support for the prediction that adaptive defenses, which are thought to be more costly to develop, would be more related to body condition than innate defenses. We discuss our findings in the context of other studies on the ontogeny of immune function.