Uncovering mechanisms of global ocean change effects on the Dungeness crab (Cancer magister) through metabolomics analysis.

The Dungeness crab is an economically and ecologically important species distributed along the North American Pacific coast. To predict how Dungeness crab may physiologically respond to future global ocean change on a molecular level, we performed untargeted metabolomic approaches on individual Dungeness crab juveniles reared in treatments that mimicked current and projected future pH and dissolved oxygen conditions. We found 94 metabolites and 127 lipids responded in a condition-specific manner, with a greater number of known compounds more strongly responding to low oxygen than low pH exposure. Pathway analysis of these compounds revealed that juveniles may respond to low oxygen through evolutionarily conserved processes including downregulating glutathione biosynthesis and upregulating glycogen storage, and may respond to low pH by increasing ATP production. Most interestingly, we found that the response of juveniles to combined low pH and low oxygen exposure was most similar to the low oxygen exposure response, indicating low oxygen may drive the physiology of juvenile crabs more than pH. Our study elucidates metabolic dynamics that expand our overall understanding of how the species might respond to future ocean conditions and provides a comprehensive dataset that could be used in future ocean acidification response studies.

Elevated CO2 impairs olfactory-mediated neural and behavioral responses and gene expression in ocean-phase coho salmon (Oncorhynchus kisutch).

Elevated concentrations of CO2 in seawater can disrupt numerous sensory systems in marine fish. This is of particular concern for Pacific salmon because they rely on olfaction during all aspects of their life including during their homing migrations from the ocean back to their natal streams. We investigated the effects of elevated seawater CO2 on coho salmon (Oncorhynchus kisutch) olfactory-mediated behavior, neural signaling, and gene expression within the peripheral and central olfactory system. Ocean-phase coho salmon were exposed to three levels of CO2 , ranging from those currently found in ambient marine water to projected future levels. Juvenile coho salmon exposed to elevated CO2 levels for 2 weeks no longer avoided a skin extract odor that elicited avoidance responses in coho salmon maintained in ambient CO2 seawater. Exposure to these elevated CO2 levels did not alter odor signaling in the olfactory epithelium, but did induce significant changes in signaling within the olfactory bulb. RNA-Seq analysis of olfactory tissues revealed extensive disruption in expression of genes involved in neuronal signaling within the olfactory bulb of salmon exposed to elevated CO2 , with lesser impacts on gene expression in the olfactory rosettes. The disruption in olfactory bulb gene pathways included genes associated with GABA signaling and maintenance of ion balance within bulbar neurons. Our results indicate that ocean-phase coho salmon exposed to elevated CO2 can experience significant behavioral impairments likely driven by alteration in higher-order neural signal processing within the olfactory bulb. Our study demonstrates that anadromous fish such as salmon may share a sensitivity to rising CO2 levels with obligate marine species suggesting a more wide-scale ecological impact of ocean acidification.

Risks of ocean acidification in the California Current food web and fisheries: ecosystem model projections.

The benefits and ecosystem services that humans derive from the oceans are threatened by numerous global change stressors, one of which is ocean acidification. Here, we describe the effects of ocean acidification on an upwelling system that already experiences inherently low pH conditions, the California Current. We used an end-to-end ecosystem model (Atlantis), forced by downscaled global climate models and informed by a meta-analysis of the pH sensitivities of local taxa, to investigate the direct and indirect effects of future pH on biomass and fisheries revenues. Our model projects a 0.2-unit drop in pH during the summer upwelling season from 2013 to 2063, which results in wide-ranging magnitudes of effects across guilds and functional groups. The most dramatic direct effects of future pH may be expected on epibenthic invertebrates (crabs, shrimps, benthic grazers, benthic detritivores, bivalves), and strong indirect effects expected on some demersal fish, sharks, and epibenthic invertebrates (Dungeness crab) because they consume species known to be sensitive to changing pH. The model's pelagic community, including marine mammals and seabirds, was much less influenced by future pH. Some functional groups were less affected to changing pH in the model than might be expected from experimental studies in the empirical literature due to high population productivity (e.g., copepods, pteropods). Model results suggest strong effects of reduced pH on nearshore state-managed invertebrate fisheries, but modest effects on the groundfish fishery because individual groundfish species exhibited diverse responses to changing pH. Our results provide a set of projections that generally support and build upon previous findings and set the stage for hypotheses to guide future modeling and experimental analysis on the effects of OA on marine ecosystems and fisheries.

Estimates of the Direct Effect of Seawater pH on the Survival Rate of Species Groups in the California Current Ecosystem.

Ocean acidification (OA) has the potential to restructure ecosystems due to variation in species sensitivity to the projected changes in ocean carbon chemistry. Ecological models can be forced with scenarios of OA to help scientists, managers, and other stakeholders understand how ecosystems might change. We present a novel methodology for developing estimates of species sensitivity to OA that are regionally specific, and applied the method to the California Current ecosystem. To do so, we built a database of all published literature on the sensitivity of temperate species to decreased pH. This database contains 393 papers on 285 species and 89 multi-species groups from temperate waters around the world. Research on urchins and oysters and on adult life stages dominates the literature. Almost a third of the temperate species studied to date occur in the California Current. However, most laboratory experiments use control pH conditions that are too high to represent average current chemistry conditions in the portion of the California Current water column where the majority of the species live. We developed estimates of sensitivity to OA for functional groups in the ecosystem, which can represent single species or taxonomically diverse groups of hundreds of species. We based these estimates on the amount of available evidence derived from published studies on species sensitivity, how well this evidence could inform species sensitivity in the California Current ecosystem, and the agreement of the available evidence for a species/species group. This approach is similar to that taken by the Intergovernmental Panel on Climate Change to characterize certainty when summarizing scientific findings. Most functional groups (26 of 34) responded negatively to OA conditions, but when uncertainty in sensitivity was considered, only 11 groups had relationships that were consistently negative. Thus, incorporating certainty about the sensitivity of species and functional groups to OA is an important part of developing robust scenarios for ecosystem projections.

Shell condition and survival of Puget Sound pteropods are impaired by ocean acidification conditions.

We tested whether the thecosome pteropod Limacina helicina from Puget Sound, an urbanized estuary in the northwest continental US, experiences shell dissolution and altered mortality rates when exposed to the high CO2, low aragonite saturation state (Ωa) conditions that occur in Puget Sound and the northeast Pacific Ocean. Five, week-long experiments were conducted in which we incubated pteropods collected from Puget Sound in four carbon chemistry conditions: current summer surface (∼460-500 µatm CO2, Ωa≈1.59), current deep water or surface conditions during upwelling (∼760 and ∼1600-1700 µatm CO2, Ωa≈1.17 and 0.56), and future deep water or surface conditions during upwelling (∼2800-3400 µatm CO2, Ωa≈0.28). We measured shell condition using a scoring regime of five shell characteristics that capture different aspects of shell dissolution. We characterized carbon chemistry conditions in statistical analyses with Ωa, and conducted analyses considering Ωa both as a continuous dataset and as discrete treatments. Shell dissolution increased linearly as aragonite saturation state decreased. Discrete treatment comparisons indicate that shell dissolution was greater in undersaturated treatments compared to oversaturated treatments. Survival increased linearly with aragonite saturation state, though discrete treatment comparisons indicated that survival was similar in all but the lowest saturation state treatment. These results indicate that, under starvation conditions, pteropod survival may not be greatly affected by current and expected near-future aragonite saturation state in the NE Pacific, but shell dissolution may. Given that subsurface waters in Puget Sound's main basin are undersaturated with respect to aragonite in the winter and can be undersaturated in the summer, the condition and persistence of the species in this estuary warrants further study.

Estimating effects of tidal power projects and climate change on threatened and endangered marine species and their food web.

Marine hydrokinetic power projects will operate as marine environments change in response to increased atmospheric carbon dioxide concentrations. We considered how tidal power development and stressors resulting from climate change may affect Puget Sound species listed under the U.S. Endangered Species Act (ESA) and their food web. We used risk tables to assess the singular and combined effects of tidal power development and climate change. Tidal power development and climate change posed risks to ESA-listed species, and risk increased with incorporation of the effects of these stressors on predators and prey of ESA-listed species. In contrast, results of a model of strikes on ESA-listed species from turbine blades suggested that few ESA-listed species are likely to be killed by a commercial-scale tidal turbine array. We applied scenarios to a food web model of Puget Sound to explore the effects of tidal power and climate change on ESA-listed species using more quantitative analytical techniques. To simulate development of tidal power, we applied results of the blade strike model. To simulate environmental changes over the next 50 years, we applied scenarios of change in primary production, plankton community structure, dissolved oxygen, ocean acidification, and freshwater flooding events. No effects of tidal power development on ESA-listed species were detected from the food web model output, but the effects of climate change on them and other members of the food web were large. Our analyses exemplify how natural resource managers might assess environmental effects of marine technologies in ways that explicitly incorporate climate change and consider multiple ESA-listed species in the context of their ecological community. Estimación de los Efectos de Proyectos de Energía de las Mareas y el Cambio Climático sobre Especies Marinas Amenazadas y en Peligro y su Red Alimentaria.

Influence of proximity to a geographical range limit on the physiology of a tropical bird.

1. Species' geographical ranges can be limited by a variety of biotic and abiotic factors. Physiological challenge in response to unsuitable environmental conditions can establish limits to geographical ranges. 2. We studied the physiology of Song Wrens (Cyphorhinus phaeocephalus) across their geographical range on the isthmus of Panama, an area characterized by a strong rainfall gradient. Wrens are common on the Caribbean slope of the isthmus where annual rainfall is greatest, but wren abundance declines towards the south as annual rainfall declines. Song Wrens are completely absent from the driest third of the isthmus. 3. We searched for the existence of a physiologically induced distribution limit by measuring body condition (an integrative measure of energy balance), hematocrit (% packed red blood cells in a given blood sample), and corticosterone levels (CORT, a steroid hormone that regulates the availability of energy and the endocrine stress response) in males and females. We caught birds by luring them into nets when they responded to playback of conspecific song. 4. Wrens living in drier habitat near the geographical range edge were significantly more likely to have abnormally low hematocrit scores. Baseline CORT levels were negatively associated with rainfall in one of our three best-fit path models, indicating potential energetic challenge in some individuals. Maximum CORT levels during a 60-min period of restraint correlated significantly only with sex, being higher in females. Birds with the poorest body condition lived at the dry end of the gradient. Birds on the wet end of the gradient responded fastest to conspecific song. 5. Environmental conditions vary across geographical ranges and may influence the physiological conditions of organisms, thereby enforcing limits to species' distributions. Highly specialized species, such as birds of the rain forest understory, may be especially susceptible to environmental variation associated with changing climatic conditions.

Seasonal and social modulation of testosterone in Costa Rican rufous-collared sparrows (Zonotrichia capensis costaricensis).

Previous work shows that most birds breeding in northern temperate regions adjust production of testosterone in response to stage of the breeding cycle and in some cases following social interactions. In contrast, prior research suggests that tropical breeding birds are less likely to modulate testosterone in response to social interactions (the propensity to increase testosterone in response to social instability is known as the challenge hypothesis). To further test the challenge hypothesis in tropical birds, we investigated whether variation in season affects reproductive condition, aggressive behavior, and social modulation of testosterone in two populations of Costa Rican rufous-collared sparrow, Zonotrichia capensis costaricensis. We conducted our study at three distinct times of year: the dry season (March and May); the veranillo, a hiatus in the rainy season (July); and the late rainy season (November). Significantly more birds were in breeding condition in the dry season than in the rainy season or veranillo. In each time period, we collected baseline testosterone samples and conducted simulated territorial intrusions (STIs). Our study shows that testosterone is modulated with season independent of breeding condition, as testosterone levels were affected by season, breeding condition, and an interaction of the two factors. Males breeding in the dry season had higher plasma testosterone levels than non-breeding males in the dry season and both breeding and non-breeding males in the veranillo and rainy season. Males did not socially modulate testosterone in any season. Aggressive behaviors expressed during STIs did not differ among seasons with the exception that males sang fewer songs during the rainy season.

Disentangling the effects of environment and life-history stage on corticosterone modulation in Costa Rican rufous-collared sparrows, Zonotrichia capensis costaricensis.

This study explores the factors that influence modulation of baseline corticosterone levels and the hypothalamo-pituitary-adrenal cortex (HPA) axis response to stress in Costa Rican rufous-collared sparrows (Zonotrichia capensis costaricensis). Individuals in our study population vary in their timing of breeding and molt. Thus, at multiple times of the year, we were able to investigate how the HPA axis changes with life-history stage (breeding, molt) and the interaction of season and life-history stage. Season explained most of the variation in corticosterone data. During the dry season, males had higher baseline corticosterone levels and both females and males had a higher stress response, regardless of breeding or molting condition. Breeding did not significantly affect corticosterone levels in either males or females. There was no direct effect of molt on corticosterone levels in males or in females, but there was a significant interaction between season and molt for male baseline corticosterone levels and HPA axis responsiveness. Baseline corticosterone levels were not correlated with body condition, suggesting that body condition is not an explanatory factor for HPA axis change. Our data indicate that environmental conditions may affect how the HPA axis in birds is modulated during different life-history stages. They also emphasize the importance of studying species with diverse life cycles and seasonality.

Impacts of frequent, acute pulses of corticosterone on condition and behavior of Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii).

Little is known about how frequent, acute stressors affect wild animals. We present two experiments conducted on captive, Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii) that explore how frequent, acute doses of corticosterone (CORT) affect condition and behavior. CORT was administered either once or three times a day to birds in pre-breeding, early-breeding, or late-breeding life-history stages. Two additional groups were included to control for the CORT delivery vehicle, DMSO, and the treatment process. Our results indicate that CORT treatment decreases condition, but that its effects are dependent on frequency and life stage. Specifically, CORT-treated birds delayed the onset of molt and had reduced body mass, flight muscle, and food consumption. CORT treatment did not affect fat stores, bile retention in the gallbladder, or the expression of migratory restlessness behavior. These results increase our understanding of the effects of frequent, acute stressors and the development of chronic stress states.

Effects of repeated, short-term, corticosterone administration on the hypothalamo-pituitary-adrenal axis of the white-crowned sparrow (Zonotrichia leucophrys gambelii).

Our knowledge of glucocorticoid actions in vertebrates comes primarily from laboratory studies, which are often conducted with little consideration of how animals experience changes in glucocorticoid secretion in natural contexts. Typically, free-living animals are exposed to acute perturbations of the environment, ranging from a few minutes to a few hours duration, with varying frequency. The cumulative effects of these perturbations and their resultant glucocorticoid surges are not well known. To investigate the possible cumulative effects of repeated, acute surges in glucocorticoid secretion, we developed an ecologically relevant methodology for treating captive white-crowned sparrows (Zonotrichia leucophrys gambelii) with corticosterone (CORT). We dissolved CORT in dimethylsulfoxide (DMSO) and administered this cocktail directly on the skin. Treatments resulted in small elevations of CORT within the physiological range. In our first experiment at the end of the breeding life stage, birds were treated three times a day (3x). Two control groups were used: one treated with DMSO 3x and one not handled nor treated. In a second study at the beginning of the breeding life stage, one group was treated once a day and a second group 3x. A DMSO-control group was used for each dosage regime. Repeated, acute administration of CORT resulted in higher baseline CORT levels and a down-regulation of the endogenous adrenocortical response to a standardized stress. Maximum CORT and plasma corticosterone binding globulin levels increased in response to the CORT treatments only at the end of the breeding season. CORT treatment did not alter adrenal size, adrenal response to ACTH, or hepatic CORT metabolism.