Regulation of gene expression is associated with tolerance of the Arctic copepod Calanus glacialis to CO2‐acidified sea water

Ocean acidification is the increase in seawater pCO₂ due to the uptake of atmospheric anthropogenic CO₂, with the largest changes predicted to occur in the Arctic seas. For some marine organisms, this change in pCO₂, and associated decrease in pH, represents a climate change‐related stressor. In this study, we investigated the gene expression patterns of nauplii of the Arctic copepod Calanus glacialis cultured at low pH levels. We have previously shown that organismal‐level performance (development, growth, respiration) of C. glacialis nauplii is unaffected by low pH. Here, we investigated the molecular‐level response to lowered pH in order to elucidate the physiological processes involved in this tolerance. Nauplii from wild‐caught C. glacialis were cultured at four pH levels (8.05, 7.9, 7.7, 7.5). At stage N6, mRNA was extracted and sequenced using RNA‐seq. The physiological functionality of the proteins identified was categorized using Gene Ontology and KEGG pathways. We found that the expression of 151 contigs varied significantly with pH on a continuous scale (93% downregulated with decreasing pH). Gene set enrichment analysis revealed that, of the processes downregulated, many were components of the universal cellular stress response, including DNA repair, redox regulation, protein folding, and proteolysis. Sodium:proton antiporters were among the processes significantly upregulated, indicating that these ion pumps were involved in maintaining cellular pH homeostasis. C. glacialis significantly alters its gene expression at low pH, although they maintain normal larval development. Understanding what confers tolerance to some species will support our ability to predict the effects of future ocean acidification on marine organisms.     

Evolution and phylogeography of Halimeda section Halimeda (Bryopsidales, Chlorophyta)

Nuclear ribosomal and plastid DNA sequences of specimens belonging to section Halimeda of the pantropical green seaweed genus Halimeda show that the group under scrutiny contains many more genetically delineable species than those recognized by classical taxonomy. Discordances between phylograms inferred from nuclear and plastid DNA sequences suggest that reticulate evolution has been involved in speciation within the clade. Nonetheless, our data do not allow ruling out certain alternative explanations for the discordances. Several pseudo-cryptic species are restricted to the margins of the generic distribution range. In a clade of H. cuneata sibling species from widely separated subtropical localities in the Indian Ocean, the South African sibling branches off first, leaving the Arabian and West Australian species as closest relatives. We hypothesize that geographic isolation of the siblings may have taken place following Pleistocene or Pliocene periods of climatic cooling during which subtropical species occupied larger distribution ranges. A more basal separation of Atlantic, Indo-Pacific, and Mediterranean species indicates vicariance. The alternative events that could have caused this vicariance are discussed.     

Expression of endogenous and exogenous growth hormone (GH) messenger (m) RNA in a GH-transgenic tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

We have previously produced transgenic fish from crosses between a wild-type female tilapia (Oreochromis niloticus) and a G1 transgenic male. This line of growth-enhanced tilapia carries a single copy of a chinook salmon (s) growth hormone (GH) gene spliced to an ocean pout antifreeze promoter (OPAFPcsGH) co-ligated to a carp -actin/lacZ reporter gene construct, integrated into the tilapia genome. Because little is known about the expression sites of transgenes, we have characterised the gene expression patterns of sGH and tilapia (t)GH in transgenic tilapia using a newly established real-time PCR to measure the absolute mRNA amounts of both hormones. The sGH gene, which was expected to be expressed mainly in liver, was also found to be expressed in other organs, such as gills, heart, brain, skeletal muscle, kidney, spleen, intestine and testes. However, in pituitary no sGH mRNA but only tGH mRNA was found. Tilapia GH mRNA in wild-type pituitary amounted to 226 ± 30 pg/g total RNA but in transgenics only to 187 ± 43 pg/g total RNA. Liver exhibited the highest level of sGH mRNA (8.3 ± 2.5 pg/g total RNA) but the extrahepatic sites expressed considerable amounts of sGH mRNA ranging from 4.1 ± 2.0 pg/g total RNA in gills to 0.2 ± 0.08 pg/g total RNA in kidney. The widespread expression of the sGH gene is assumed to be due to the tissue specificity of the type III AFP gene promoter. It is assumed that our transgenic experiments, which in contrast to some other approaches caused no obvious organ abnormalities, mimick the GH expression during ontogeny. Because sGH mRNA is expressed both in liver and in extrahepatic sites it may not only promote secretion and release of liver-derived (endocrine) IGF-I leading to an overall growth enhancement but also stimulate IGF-I expression within the different organs in a paracrine/autocrine manner and, thus, further promote organ growth.     

Pioneering Interdisciplinary Research Initiatives for Oceans and Human Health

Following the release of the report From Monsoons to Microbes by the National Research Council in 1999, efforts began to promote federal sponsorship of research and education in a new scientific discipline focusing on how the ocean affects human health. The National Institute of Environmental Health Sciences (NIEHS) and the National Science Foundation (NSF) initiated a joint program to establish and sustain several research Centers for Oceans and Human Health (COHH) at nonfederal institutions. Shortly thereafter, the National Oceanic and Atmospheric Administration (NOAA) mounted a similar initiative to establish intramural centers at existing NOAA facilities as well as an extramural grants program. This profile reviews the history and current state of these developments. The statements and opinions in this report are those of the authors only and do not necessarily represent the position of, or imply commitments by, any agency of the United States Government.     

Spatial variability overwhelms seasonal patterns in bacterioplankton communities across a river to ocean gradient

Few studies of microbial biogeography address variability across both multiple habitats and multiple seasons. Here we examine the spatial and temporal variability of bacterioplankton community composition of the Columbia River coastal margin using 16S amplicon pyrosequencing of 300 water samples collected in 2007 and 2008. Communities separated into seven groups (ANOSIM, P<0.001): river, estuary, plume, epipelagic, mesopelagic, shelf bottom (depth<350 m) and slope bottom (depth>850 m). The ordination of these samples was correlated with salinity (ρ=−0.83) and depth (ρ=−0.62). Temporal patterns were obscured by spatial variability among the coastal environments, and could only be detected within individual groups. Thus, structuring environmental factors (for example, salinity, depth) dominate over seasonal changes in determining community composition. Seasonal variability was detected across an annual cycle in the river, estuary and plume where communities separated into two groups, early year (April–July) and late year (August–Nov), demonstrating annual reassembly of communities over time. Determining both the spatial and temporal variability of bacterioplankton communities provides a framework for modeling these communities across environmental gradients from river to deep ocean.     

Adult exposure influences offspring response to ocean acidification in oysters

It is essential to predict the impact of elevated Pco₂ on marine organisms and habitats to anticipate the severity and consequences of future ocean chemistry change. Despite the importance of carry‐over effects in the evolutionary history of marine organisms, few studies have considered links between life‐history stages when determining how marine organisms will respond to elevated Pco₂, and none have considered the link between adults and their offspring. Herein, we exposed adults of wild and selectively bred Sydney rock oysters, Saccostrea glomerata to elevated Pco₂ during reproductive conditioning and measured the development, growth and survival response of their larvae. We found that elevated Pco₂ had a negative impact on larvae of S. glomerata causing a reduction in growth, rate of development and survival. Exposing adults to elevated Pco₂ during reproductive conditioning, however, had positive carry‐over effects on larvae. Larvae spawned from adults exposed to elevated Pco₂ were larger and developed faster, but displayed similar survival compared with larvae spawned from adults exposed to ambient Pco₂. Furthermore, selectively bred larvae of S. glomerata were more resilient to elevated Pco₂ than wild larvae. Measurement of the standard metabolic rate (SMR) of adult S. glomerata showed that at ambient Pco₂, SMR is increased in selectively bred compared with wild oysters and is further increased during exposure to elevated Pco₂. This study suggests that sensitive marine organisms may have the capacity to acclimate or adapt to elevated Pco₂ over the next century and a change in energy turnover indicated by SMR may be a key process involved.     

Acclimation to ocean acidification during long‐term CO2 exposure in the cold‐water coral Lophelia pertusa

Ocean acidity has increased by 30% since preindustrial times due to the uptake of anthropogenic CO₂ and is projected to rise by another 120% before 2100 if CO₂ emissions continue at current rates. Ocean acidification is expected to have wide‐ranging impacts on marine life, including reduced growth and net erosion of coral reefs. Our present understanding of the impacts of ocean acidification on marine life, however, relies heavily on results from short‐term CO₂ perturbation studies. Here, we present results from the first long‐term CO₂ perturbation study on the dominant reef‐building cold‐water coral Lophelia pertusa and relate them to results from a short‐term study to compare the effect of exposure time on the coral's responses. Short‐term (1 week) high CO₂ exposure resulted in a decline of calcification by 26–29% for a pH decrease of 0.1 units and net dissolution of calcium carbonate. In contrast, L. pertusa was capable to acclimate to acidified conditions in long‐term (6 months) incubations, leading to even slightly enhanced rates of calcification. Net growth is sustained even in waters sub‐saturated with respect to aragonite. Acclimation to seawater acidification did not cause a measurable increase in metabolic rates. This is the first evidence of successful acclimation in a coral species to ocean acidification, emphasizing the general need for long‐term incubations in ocean acidification research. To conclude on the sensitivity of cold‐water coral reefs to future ocean acidification further ecophysiological studies are necessary which should also encompass the role of food availability and rising temperatures.     

Synergistic effects of ocean acidification and warming on overwintering pteropods in the Arctic

Ocean acidification and warming will be most pronounced in the Arctic Ocean. Aragonite shell‐bearing pteropods in the Arctic are expected to be among the first species to suffer from ocean acidification. Carbonate undersaturation in the Arctic will first occur in winter and because this period is also characterized by low food availability, the overwintering stages of polar pteropods may develop into a bottleneck in their life cycle. The impacts of ocean acidification and warming on growth, shell degradation (dissolution), and mortality of two thecosome pteropods, the polar Limacina helicina and the boreal L. retroversa, were studied for the first time during the Arctic winter in the Kongsfjord (Svalbard). The abundance of L. helicina and L. retroversa varied from 23.5 to 120 ind m^?2 and 12 to 38 ind m^?2, and the mean shell size ranged from 920 to 981 μm and 810 to 823 μm, respectively. Seawater was aragonite‐undersaturated at the overwintering depths of pteropods on two out of ten days of our observations. A 7‐day experiment [temperature levels: 2 and 7 °C, pCO₂ levels: 350, 650 (only for L. helicina) and 880 μatm] revealed a significant pCO₂ effect on shell degradation in both species, and synergistic effects between temperature and pCO₂ for L. helicina. A comparison of live and dead specimens kept under the same experimental conditions indicated that both species were capable of actively reducing the impacts of acidification on shell dissolution. A higher vulnerability to increasing pCO₂ and temperature during the winter season is indicated compared with a similar study from fall 2009. Considering the species winter phenology and the seasonal changes in carbonate chemistry in Arctic waters, negative climate change effects on Arctic thecosomes are likely to show up first during winter, possibly well before ocean acidification effects become detectable during the summer season.     

Distribution and diversity of a protist predator Cryothecomonas (Cercozoa) in Arctic marine waters

Heterotrophic nanoflagellates (HNFs) are key components in microbial food webs, potentially influencing community composition via top-down control of their favored prey or host. Marine cercozoan Cryothecomonas species are parasitoid and predatory HNFs that have been reported from ice, sediments, and the water column. Although Cryothecomonas is frequently reported from Arctic and subarctic seas, factors determining its occurrence are not known. We investigated the temporal and geographic distribution of Cryothecomonas in Canadian Arctic seas during the summer and autumn periods from 2006 to 2010. We developed a Cryothecomonas-specific fluorescent in situ hybridization (FISH) probe targeting ribosomal 18S rRNA to estimate cell concentrations in natural and manipulated samples. Comparison of simple and partial correlation coefficients showed that salinity, depth, and overall community biomass are important factors determining Cryothecomonas abundance. We found no evidence of parasitism in our samples. Hybridized cells included individuals smaller than any formally described Cryothecomonas, suggesting the presence of novel taxa or unknown life stages in this genus. A positive relationship between Cryothecomonas abundance and ice and meltwater suggests that it is a sensitive indicator of ice melt in Arctic water columns.     

CARBON‐USE STRATEGIES IN MACROALGAE: DIFFERENTIAL RESPONSES TO LOWERED PH AND IMPLICATIONS FOR OCEAN ACIDIFICATION1

Ocean acidification (OA) is a reduction in oceanic pH due to increased absorption of anthropogenically produced CO₂. This change alters the seawater concentrations of inorganic carbon species that are utilized by macroalgae for photosynthesis and calcification: CO₂ and HCO₃^? increase; CO₃^2? decreases. Two common methods of experimentally reducing seawater pH differentially alter other aspects of carbonate chemistry: the addition of CO₂ gas mimics changes predicted due to OA, while the addition of HCl results in a comparatively lower [HCO₃^?]. We measured the short‐term photosynthetic responses of five macroalgal species with various carbon‐use strategies in one of three seawater pH treatments: pH 7.5 lowered by bubbling CO₂ gas, pH 7.5 lowered by HCl, and ambient pH 7.9. There was no difference in photosynthetic rates between the CO₂, HCl, or pH 7.9 treatments for any of the species examined. However, the ability of macroalgae to raise the pH of the surrounding seawater through carbon uptake was greatest in the pH 7.5 treatments. Modeling of pH change due to carbon assimilation indicated that macroalgal species that could utilize HCO₃^? increased their use of CO₂ in the pH 7.5 treatments compared to pH 7.9 treatments. Species only capable of using CO₂ did so exclusively in all treatments. Although CO₂ is not likely to be limiting for photosynthesis for the macroalgal species examined, the diffusive uptake of CO₂ is less energetically expensive than active HCO₃^? uptake, and so HCO₃^?‐using macroalgae may benefit in future seawater with elevated CO₂.