Integrative biological analyses of responses to food deprivation reveal resilience mechanisms in sea urchin larvae.

A fundamental question in ecology is how organisms survive food deprivation. In the ocean, climate change is impacting the phenology of food availability for early life-history stages of animals. In this study, we undertook an integrative analysis of larvae of the sea urchin Strongylocentrotus purpuratus-an important keystone species in marine ecology and a molecular biological model organism in developmental biology. Specifically, to identify the mechanisms of resilience that maintain physiological state and the ability of organisms to recover from food deprivation, a suite of molecular biological, biochemical, physiological and whole organism measurements was completed. Previous studies focused on the importance of energy reserves to sustain larvae during periods of food deprivation. We show, however, that utilization of endogenous energy reserves only supplied 15% of the metabolic requirements of long-term survival (up to 22 days) in the absence of particulate food. This large energy gap was not supplied by larvae feeding on bacteria. Estimates of larval ability to transport dissolved organic matter directly from seawater showed that such substrates could fully supply metabolic needs. Integrative approaches allowed for filtering of gene expression signatures, linked with gene network analyses and measured biochemical and physiological traits, to identify biomarkers of resilience. We identified 14 biomarkers related to nutrition-responsive gene expression, of which a specific putative amino acid transporter gene was quantified in a single larva experiencing continuous nutritional stress. Advances in applications of gene expression technologies offer novel approaches to determine the physiological state of marine larval forms in ecological settings undergoing environmental change.

Thermal sensitivities of respiration and protein synthesis differ among larval families of the Pacific oyster, Crassostrea gigas.

Understanding the mechanisms of biological responses to environmental change is a central theme in comparative and evolutionary physiology. Here, we analyzed variation in physiological responses to temperature, using 21 full-sibling larval families of the Pacific oyster, Crassostrea gigas. Pedigrees were confirmed with genetic markers for adult broodstock obtained from our breeding program. From these 21 larval families, 41 determinations of thermal sensitivity (Q10 values) were assayed for larvae of different sizes. For respiration, thermal sensitivity was consistent within a larval family during growth, but showed significant differences among families. Different Q10 values were evident among 21 larval families, with family accounting for 87% of variation. Specifically, four larval families maintained an increased thermal sensitivity for respiration (Q10 of 3). This physiology would confer resilience to rising temperature by matching the increased energy demand of protein synthesis (Q10 of 3 previously reported). For protein synthesis, differences in Q10 values were also observed. Notably, a family was identified that had a decreased thermal sensitivity for protein synthesis (Q10 of 1.7 cf. Q10 of 3 for other families), conferring an optimal energy allocation with rising temperature. Different thermal sensitivities across families for respiration (energy supply) and protein synthesis (energy demand) were integrated into models of energy allocation at the whole-organism level. The outcome of these analyses provides insights into the physiological bases of optimal energy allocation with rising temperature. These transgenerational (egg-to-egg) experiments highlight approaches to dissect components of phenotypic variance to address long-standing questions of genetic adaptation and physiological resilience to environmental change.

Harmful algal blooms: A climate change co-stressor in marine and freshwater ecosystems.

Marine and freshwater ecosystems are warming, acidifying, and deoxygenating as a consequence of climate change. In parallel, the impacts of harmful algal blooms (HABs) on these ecosystems are intensifying. Many eutrophic habitats that host recurring HABs already experience thermal extremes, low dissolved oxygen, and low pH, making these locations potential sentinel sites for conditions that will become more common in larger-scale systems as climate change accelerates. While studies of the effects of HABs or individual climate change stressors on aquatic organisms have been relatively common, studies assessing their combined impacts have been rare. Those doing so have reported strong species- and strain-specific interactions between HAB species and climate change co-stressors yielding outcomes for aquatic organisms that could not have been predicted based on investigations of these factors individually. This review provides an ecological and physiological framework for considering HABs as a climate change co-stressor and considers the consequences of their combined occurrence for coastal ecosystems. This review also highlights critical gaps in our understanding of HABs as a climate change co-stressor that must be addressed in order to develop management plans that adequately protect fisheries, aquaculture, aquatic ecosystems, and human health. Ultimately, incorporating HAB species into experiments and monitoring programs where the effects of multiple climate change stressors are considered will provide a more ecologically relevant perspective of the structure and function of marine ecosystems in future, climate-altered systems.

Ocean warming along temperate western boundaries of the Northern Hemisphere promotes an expansion of Cochlodinium polykrikoides blooms.

Since the early 1990s, ocean temperatures have increased and blooms of the icthyotoxic dinoflagellate Cochlodinium polykrikoides (a.k.a. Margalefidinium polykrikoides) have become more widespread across the Northern Hemisphere. This study used high-resolution (1-30 km), satellite-based sea surface temperature records since 1982 to model trends in growth and bloom season length for strains of C. polykrikoides inhabiting North American and East Asian coastlines to understand how warming has altered blooms in these regions. Methods provided approximately 180× greater spatial resolution than previous studies of the impacts of warming on harmful algae, providing novel insight into near shore, coastal environments. Along the US East Coast, significant increases in potential growth rates and bloom season length for North American ribotypes were observed with bloom-favourable conditions becoming established earlier and persisting longer from Chesapeake Bay through Cape Cod, areas where blooms have become newly established and/or intensified this century. Within the Sea of Japan, modelled mean potential growth rates and bloom season length of East Asian ribotypes displayed a significant positive correlation with rising sea surface temperatures since 1982, a period during which observed maximal cell densities of C. polykrikoides blooms have significantly increased. Results suggest that warming has contributed, in part, to altering the phenology of C. polykrikoides populations, potentially expanding its realized niche in temperate zones of the Northern Hemisphere.

The harmful algae, Cochlodinium polykrikoides and Aureococcus anophagefferens, elicit stronger transcriptomic and mortality response in larval bivalves (Argopecten irradians) than climate change stressors.

Global ocean change threatens marine life, yet a mechanistic understanding of how organisms are affected by specific stressors is poorly understood. Here, we identify and compare the unique and common transcriptomic responses of an organism experiencing widespread fisheries declines, Argopecten irradians (bay scallop) exposed to multiple stressors including high pCO2, elevated temperature, and two species of harmful algae, Cochlodinium (aka Margalefidinium) polykrikoides and Aureococcus anophagefferens using high-throughput sequencing (RNA-seq). After 48 hr of exposure, scallop transcriptomes revealed distinct expression profiles with larvae exposed to harmful algae (C. polykrikoides and A. anophagefferens) displaying broader responses in terms of significantly and differentially expressed (DE) transcripts (44,922 and 4,973; respectively) than larvae exposed to low pH or elevated temperature (559 and 467; respectively). Patterns of expression between larvae exposed to each harmful algal treatment were, however, strikingly different with larvae exposed to A. anophagefferens displaying large, significant declines in the expression of transcripts (n = 3,615; 87% of DE transcripts) whereas exposure to C. polykrikoides increased the abundance of transcripts, more than all other treatments combined (n = 43,668; 97% of DE transcripts). Larvae exposed to each stressor up-regulated a common set of 21 genes associated with protein synthesis, cellular metabolism, shell growth, and membrane transport. Larvae exposed to C. polykrikoides displayed large increases in antioxidant-associated transcripts, whereas acidification-exposed larvae increased abundance of transcripts associated with shell formation. After 10 days of exposure, each harmful algae caused declines in survival that were significantly greater than all other treatments. Collectively, this study reveals the common and unique transcriptional responses of bivalve larvae to stressors that promote population declines within coastal zones, providing insight into the means by which they promote mortality as well as traits possessed by bay scallops that enable potential resistance.

In vivo characterization of bivalve larval shells: a confocal Raman microscopy study.

In vivo confocal Raman microscopy (CRM), polarized light microscopy and Fourier transform infrared spectroscopy (FTIR) were used to determine if a significant amount of amorphous calcium carbonate (ACC) exists within larval shells of Baltic mytilid mussels (Mytilus edulis-like) and whether the amount of ACC varies during larval development. No evidence for ACC was found from the onset of shell deposition at 21 h post-fertilization (hpf) until 48 hpf. Larval Mytilus shells were crystalline from 21 hpf onwards and exhibited CRM and FTIR peaks characteristic of aragonite. Prior to shell deposition at 21 hpf, no evidence for carbonates was observed through in vivo CRM. We further analysed the composition of larval shells in three other bivalve species, Mercenaria mercenaria, Crassostrea gigas and Crassostrea virginica and observed no evidence for ACC, which is in contrast to previous work on the same species. Our findings indicate that larval bivalve shells are composed of crystalline aragonite and we demonstrate that conflicting results are related to sub-optimal measurements and misinterpretation of CRM spectra. Our results demonstrate that the common perception that ACC generally occurs as a stable and abundant precursor during larval bivalve calcification needs to be critically reviewed.

Transgenerational exposure of North Atlantic bivalves to ocean acidification renders offspring more vulnerable to low pH and additional stressors.

While early life-stage marine bivalves are vulnerable to ocean acidification, effects over successive generations are poorly characterized. The objective of this work was to assess the transgenerational effects of ocean acidification on two species of North Atlantic bivalve shellfish, Mercenaria mercenaria and Argopecten irradians. Adults of both species were subjected to high and low pCO2 conditions during gametogenesis. Resultant larvae were exposed to low and ambient pH conditions in addition to multiple, additional stressors including thermal stress, food-limitation, and exposure to a harmful alga. There were no indications of transgenerational acclimation to ocean acidification during experiments. Offspring of elevated pCO2-treatment adults were significantly more vulnerable to acidification as well as the additional stressors. Our results suggest that clams and scallops are unlikely to acclimate to ocean acidification over short time scales and that as coastal oceans continue to acidify, negative effects on these populations may become compounded and more severe.

Ocean warming since 1982 has expanded the niche of toxic algal blooms in the North Atlantic and North Pacific oceans.

Global ocean temperatures are rising, yet the impacts of such changes on harmful algal blooms (HABs) are not fully understood. Here we used high-resolution sea-surface temperature records (1982 to 2016) and temperature-dependent growth rates of two algae that produce potent biotoxins, Alexandrium fundyense and Dinophysis acuminata, to evaluate recent changes in these HABs. For both species, potential mean annual growth rates and duration of bloom seasons significantly increased within many coastal Atlantic regions between 40°N and 60°N, where incidents of these HABs have emerged and expanded in recent decades. Widespread trends were less evident across the North Pacific, although regions were identified across the Salish Sea and along the Alaskan coastline where blooms have recently emerged, and there have been significant increases in the potential growth rates and duration of these HAB events. We conclude that increasing ocean temperature is an important factor facilitating the intensification of these, and likely other, HABs and thus contributes to an expanding human health threat.

Hypoxia and acidification have additive and synergistic negative effects on the growth, survival, and metamorphosis of early life stage bivalves.

Low oxygen zones in coastal and open ocean ecosystems have expanded in recent decades, a trend that will accelerate with climatic warming. There is growing recognition that low oxygen regions of the ocean are also acidified, a condition that will intensify with rising levels of atmospheric CO2. Presently, however, the concurrent effects of low oxygen and acidification on marine organisms are largely unknown, as most prior studies of marine hypoxia have not considered pH levels. We experimentally assessed the consequences of hypoxic and acidified water for early life stage bivalves (bay scallops, Argopecten irradians, and hard clams, Mercenaria mercenaria), marine organisms of significant economic and ecological value and sensitive to climate change. In larval scallops, experimental and naturally-occurring acidification (pH, total scale  = 7.4-7.6) reduced survivorship (by >50%), low oxygen (30-50 µM) inhibited growth and metamorphosis (by >50%), and the two stressors combined produced additively negative outcomes. In early life stage clams, however, hypoxic waters led to 30% higher mortality, while acidified waters significantly reduced growth (by 60%). Later stage clams were resistant to hypoxia or acidification separately but experienced significantly (40%) reduced growth rates when exposed to both conditions simultaneously. Collectively, these findings demonstrate that the consequences of low oxygen and acidification for early life stage bivalves, and likely other marine organisms, are more severe than would be predicted by either individual stressor and thus must be considered together when assessing how ocean animals respond to these conditions both today and under future climate change scenarios.

Bioaccumulation and depuration of brevetoxins in the eastern oyster (Crassostrea virginica) and the northern quahog (= hard clam, Mercenaria mercenaria).

The eastern oyster (Crassostrea virginica) and northern quahog (= hard clam, Mercenaria mercenaria) are two species of economic and ecological significance in east coast waters of the United States and the Gulf of Mexico. Commercial industries for these species, especially within the state of Florida, are significant. The current study was undertaken to build upon the already established body of knowledge surrounding effects of the toxic dinoflagellate Karenia brevis on shellfish, to provide an understanding of the kinetics of brevetoxins within shellfish tissues, and to provide an estimate of brevetoxin retention times in these shellfish after a bloom event. Individual clams and oysters were exposed to the toxic dinoflagellate, K. brevis at a bloom concentration of 5 × 10(5) cells·L(-1) for eight days and then transferred to filtered water for depuration. Individuals were sampled periodically to determine depuration rates. Concentrations of brevetoxins (and/or their metabolites measured as PbTx-3 equivalent) in tissues were determined using an Enzyme Linked Immunosorbent Assay (ELISA). After five days of exposure, brevetoxin levels in tissues of both species reached concentrations well above the regulatory limit of 800 ng g(-1) (Pb-TX3 equivalent). Averaged concentration of brevetoxins in clams was 1000 ng g(-1), while the oysters averaged 1986 ng g(-1). After two weeks of depuration, tissue concentrations in both species were below regulatory levels with clams averaging ~204 ng g(-1) and oysters averaging ~437 ng g(-1). Toxins (or their metabolities) remained detectable in both clams (139 days) and oysters (82 days) for the duration of the experiment.