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Human activity has increased concentrations of endocrine-disrupting contaminants (EDCs) in many ecological systems. Many EDCs are xenoestrogens, which imitate naturally derived estrogen like estradiol 17-β (E2). These pollutants can critically affect a broad range of biological functions, particularly in organisms inhabiting aquatic environments. E2 and associated receptors are involved in regulating innate immune responses, where documentation of exogenous E2 on immune parameters is important for understanding health consequences. In this study, we explore the impact of environmentally relevant concentrations of E2 on circulating glucocorticoid levels and several innate immune parameters in hatchling American alligators (Alligator mississippiensis). Twenty-three hatchling alligators were randomly placed in one of three groups that differed in dietary E2 concentration: control (no E2 exposure), low E2 (0.5 μg/kg E2), or high E2 (1 μg/kg E2) for 10 weeks. Following this period, several biomarkers were quantified to monitor the impact of E2: growth, change in body condition, white blood cell (WBC) counts, glucocorticoid levels, and general antibody response. Blood E2 concentrations were greater in individuals exposed to E2, but plasma corticosterone levels were reduced among the experimental groups. Morphology, growth, and immune parameters of E2 exposed animals did not differ from controls. These results suggest that acute exposure to increased environmental estrogen concentrations may alter plasma hormone concentrations but have little to no impact on immediate morphology or immune responses. Future studies may expand on this by monitoring biomarkers in wild populations across time, which will provide insight into how different ontogenetic stages are impacted by environmental contaminants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

The cane toad (Rhinella marina) is an invasive amphibian in several parts of the world. Much of the research performed on assessing the dispersal potential of invasive species has focused immunity. Invaders are predicted to rely less on pro-inflammatory immunity, allowing them to allocate energy to dispersal. Elevated stress may play a role in regulation of immune responses used by invasive species. RNA sequencing of spleen tissue from cane toads subjected to an acute LPS challenge revealed genes coding for cytokines involved in typical innate responses such as phagocytic cell recruitment, extravasation, inflammation, and lymphocyte differentiation were significantly upregulated, while toads receiving transdermal application of corticosterone in addition to an LPS injection showed downregulation of genes involved with cell mediated immunity. These results indicate hormonal changes associated with acute stress may alter investment into mounting cell-mediated or humoral responses while allowing for prolonged phagocytic innate responses in this invasive species., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The possible trade-off between the roles of glucocorticoids as facilitators of energy substrate mobilization and neural inhibitors of sexual behavior during breeding season is under debate. We studied the relationship between calling and territorial behavior with plasma levels of corticosterone (CORT) and plasma levels of testosterone (T) across the breeding season of Hypsiboas faber, a large and territorial Neotropical treefrog. We investigated these relationships through focal observations of males calling naturally, followed by blood sampling for hormonal radioimmunoassay. We additionally used an experimental approach, which consisted of broadcasting recorded advertisement calls for 10 min to simulate an invasion in the territory of the focal subjects, followed by behavioral observation and blood sampling for hormonal radioimmunoassay. Results showed a pattern of co-variation between CORT and T across the breeding season. Furthermore, individual variation in CORT and T was related to different aspects of behavior: individuals with higher CORT showed higher calling rates, and individuals with higher steroid levels, mainly T, showed higher responsivity to social stimulation by other males in the chorus. Experimental simulation of territorial intrusion by using playback of advertisement calls of this species did not elicit consistent changes in agonistic behavior and CORT, but decreased T in focal males., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Graptemys flavimaculata, the yellow-blotched map turtle, is a long-lived, threatened, species, endemic to the Pascagoula River drainage in Mississippi. During the 1980s, one branch of the drainage (i.e. the Leaf River) was impacted by effluent from a wood pulp processing plant known to contain endocrine disrupters. A decade later, we examined seasonal reproductive parameters (i.e. monthly plasma estradiol-17beta (E(2)), testosterone (T), vitellogenin (VTG) and follicular development) in adult female turtles from historically polluted and reference sites in the drainage to determine if legacy exposure to pollution impacts reproduction . We found no seasonal patterns in E(2) or T and these patterns did not differ between sites. However, E(2) differed significantly among ovarian stages for the reference, but not pollutant exposed females. A significantly greater percentage of reference site females were able to produce a second clutch than females from the historically polluted site (50% and 17%). Additionally, there was a significant positive correlation between E(2) with VTG levels for reference, but not pollutant exposed females. Body and yolk tissue contaminant analysis indicated that exposure to pollutants is presently minimal and unlikely the cause of the reproductive differences observed between sites; instead, differences are potentially due to exposure history.

Coal combustion residues (CCRs) are documented to negatively impact oral morphology, growth, and development in larval amphibians. It is currently unclear what physiological mechanisms may mediate these effects. Corticosterone, a glucocorticoid hormone, is a likely mediator because when administered exogenously it, like CCRs, also negatively influences oral morphology, growth, and development in larval amphibians. In an attempt to identify if corticosterone mediates these effects, we raised larval Southern Leopard Frogs, Rana sphenocephala, on either sand or CCR substrate and documented effects of sediment type on whole body corticosterone, oral morphology, and time to and mass at key metamorphic stages. Coal combustion residue treated tadpoles contained significantly more corticosterone than controls throughout metamorphosis. However, significantly more oral abnormalities occurred early in metamorphosis when differences in corticosterone levels between treatments were minimal. Overall, CCR-treated tadpoles took significantly more time to transition between key stages and gained less mass between stages than controls, but these differences between treatments decreased during later stages when corticosterone differences between treatments were greatest. Our results suggest endogenous increase in corticosterone content and its influence on oral morphology, growth and development is more complex than previously thought.

Avian egg yolks are made up of complex mixtures of physiologically relevant substances. Androgens, in particular, accumulate in yolks in variable amounts based on social and environmental conditions experienced by laying females, and using experimental elevations of yolk androgen content, researchers have unveiled potent physiological and behavioral effects on offspring. These patterns and effects are exciting in an adaptive context, as the transfer of endogenously-produced substances such as androgens to egg yolks may allow fine manipulation of offspring phenotype to maximize reproductive success. However, to gain an in-depth understanding of how yolk androgens function in an adaptive context, we must first understand the complex entanglement of physiological and endocrine interrelationships that change after exposure to yolk androgens. Here, we take a comparative approach towards a discussion of how yolk androgens can simultaneously affect multiple body systems within developing birds, ultimately resulting in the large-scale phenotypic endpoints that may represent adaptive consequences of exposure to elevated levels of yolk androgens.

Southern toads (Bufo terrestris) are found in coal fly ash collection basins associated with coal-burning electrical power plants. These basins contain large amounts of trace metals and organisms found in these basins are known to accumulate large quantities of metals. Studies on a variety of organisms exposed to trace metals found that they experience a significant increase in standard metabolic rate. We experimentally exposed southern toads to metal-contaminated sediment and food and measured changes in standard and exercise metabolic rates as well as changes in body, liver and muscle mass, blood glucose, and corticosterone. We found that toads exposed to trace metal contamination gained significantly less mass (18.3 %) than control toads (31.3%) when food was limited and experienced significantly decreased RQ after exercise. However, contaminated toads did not experience changes in standard (x control = 0.114 +/- 0.016 mL O2 g(-1) h(-1); x (ash)=0.109 +/- 0.013 mL O2 g(-1) h(-1), p = 0.08) or exercise metabolic rates (x control = 0.53 +/- 0.06 mL O2 g(-1) h(-1); x ash =0.44 +/- 0.04 mL O2 g(-1) h(-1), p = 0.47) plasma glucose levels (p = 0.6), and hepatic or muscle percentage indices (p > 0.2 in all cases) whether food was limited or not.

From May to September of 1998, we collected monthly plasma samples from male yellow-blotched map turtles captured at two sites in the Pascagoula River drainage, Mississippi. One site (Vancleave) has a documented history of pollution from industrial sources (principally 2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD). Fish consumption advisories at the Vancleave site were lifted in 1996 and current impacts appear minimal. However, the yellow-blotched map turtle, a federally protected species, continues to decline in numbers. To determine if endocrine disruption could be a factor in the low reproductive rates observed in Vancleave turtles, we examined levels of plasma testosterone (T) and estradiol-17beta (E(2)) from males at this site and a second site (Leaksville), which has no known source of industrial pollution. Plasma was also tested for vitellogenin (VTG), which, in males, can be a biomarker of exposure to estrogenic contaminants. No males had detectable plasma VTG nor did mean monthly E(2) levels differ between sites. However, 10% of males from the historically polluted site were found to have high levels of E(2) (equivalent to levels found in females) and T was significantly lower for males captured at this site for 3 of 5 months. Our data suggest that the current impact of contaminants on reproduction in this population is limited. However, a portion of the population may have been affected developmentally, as represented by differences in reproductive parameters detected between sites.

To assess the responsiveness of the interrenal axis to stress, we injected toads exposed to coal combustion wastes and toads from an unpolluted reference site with adrenocorticotropic hormone (ACTH), as well as the vehicle alone (saline). Initial circulating levels of corticosterone in toads captured at the polluted area were significantly higher than levels in toads from the reference site. Corticosterone levels in toads from the polluted site remained high even after 2 weeks of laboratory acclimation and injection with saline. The results may suggest disruption of hepatic enzymes responsible for the metabolic clearance of steroid hormones. Injection of toads from the polluted site with ACTH had no effect on plasma corticosterone levels, whereas a similar treatment of toads from the reference site stimulated a marked increase in corticosterone. Our study provides evidence that toads exposed to coal combustion wastes may be less efficient at responding to additional environmental stressors.