Temperature tolerance of different larval stages of the spider crab Hyas araneus exposed to elevated seawater PCO2

Exposure to elevated seawater PCO2 limits the thermal tolerance of crustaceans but the underlying mechanisms have not been comprehensively explored. Larval stages of crustaceans are even more sensitive to environmental hypercapnia and possess narrower thermal windows than adults. In a mechanistic approach, we analysed the impact of high seawater CO2 on parameters at different levels of biological organization, from the molecular to the whole animal level. At the whole animal level we measured oxygen consumption, heart rate and activity during acute warming in zoea and megalopa larvae of the spider crab Hyas araneus exposed to different levels of seawater PCO2. Furthermore, the expression of genes responsible for acid-base regulation and mitochondrial energy metabolism, and cellular responses to thermal stress (e.g. the heat shock response) was analysed before and after larvae were heat shocked byrapidly raising the seawater temperature from 10°C rearing temperature to 20°C. Zoea larvae showed a high heat tolerance, which decreased at elevated seawater PCO2, while the already low heat tolerance of megalopa larvae was not limited further by hypercapnic exposure. There was a combined effect of elevated seawater CO2 and heat shock in zoea larvae causing elevated transcript levels of heat shock proteins. In all three larval stages, hypercapnic exposure elicited an up-regulation of genes involved in oxidative phosphorylation, which was, however, not accompanied by increased energetic demands. The combined effect of seawater CO2 and heat shock on the gene expression of heat shock proteins reflects the downward shift in thermal limits seen on the whole animal level and indicates an associated capacity to elicit passive thermal tolerance. The up-regulation of genes involved in oxidative phosphorylation might compensate for enzyme activities being lowered through bicarbonate inhibition and maintain larval standard metabolic rates at high seawater CO2 levels. The present study underlines the necessity to align transcriptomic data with physiological responses when addressing mechanisms affected by an interaction of elevated seawater PCO2 and temperature extremes.

Mesocosm experiment 2012 on warming and acidification effects on phytoplankton biomass and chemical composition

We investigated the impacts of predicted ocean acidification and future warming on the quantity and nutritional quality of a natural phytoplankton autumn bloom in a mesocosm experiment. Since the effects of CO2-enrichment and temperature have usually been studied independently, we were also interested in the interactive effects of both aspects of climate change. Therefore, we used a factorial design with two temperature and two acidification levels in a mesocosm experiment with a Baltic Sea phytoplankton community. Our results show a significant time-dependent influence of warming on phytoplankton carbon, chlorophyll a as well as POC. Phytoplankton carbon for instance decreased by more than a half with increasing temperature at bloom time. Additionally, elemental carbon to phosphorus ratios (C:P) increased significantly by approximately 5-8 % under warming. Impacts of CO2 or synergetic effects of warming and acidification could not be detected. We suggest that temperature-induced stronger grazing pressure was responsible for the significant decline in phytoplankton biomass. Our results suggest that biological effects of warming on Baltic Sea phytoplankton are considerable and will likely have fundamental consequences for the trophic transfer in the pelagic food-web.

Potential impact of DOM accumulation on fCO2 and carbonate ion computations in ocean acidification experiments

Culture and mesocosm experiments are often carried out under high initial nutrient concentrations, yielding high biomass concentrations that in turn often lead to a substantial build-up of DOM. In such experiments, DOM can reach concentrations much higher than typically observed in the open ocean. To the extent that DOM includes organic acids and bases, it will contribute to the alkalinity of the seawater contained in the experimental device. Our analysis suggests that whenever substantial amounts of DOM are produced during the experiment, standard computer programmes used to compute CO2 fugacity can underestimate true fCO2 significantly when the computation is based on AT and CT. Unless the effect of DOM-alkalinity can be accounted for, this might lead to significant errors in the interpretation of the system under consideration with respect to the experimentally applied CO2 perturbation. Errors in the inferred fCO2 can misguide the development of parameterisations used in simulations with global carbon cycle models in future CO2-scenarios. Over determination of the CO2-system in experimental ocean acidification studies is proposed to safeguard against possibly large errors in estimated fCO2.

Biogeochemistry of mesocosm experiments in the tropical Pacific and Atlantic Ocean

Oxygen-deficient waters in the ocean, generally referred to as oxygen minimum zones (OMZ), are expected to expand as a consequence of global climate change. Poor oxygenation is promoting microbial loss of inorganic nitrogen (N) and increasing release of sediment-bound phosphate (P) into the water column. These intermediate water masses, nutrient-loaded but with an N deficit relative to the canonical N:P Redfield ratio of 16:1, are transported via coastal upwelling into the euphotic zone. To test the impact of nutrient supply and nutrient stoichiometry on production, partitioning and elemental composition of dissolved (DOC, DON, DOP) and particulate (POC, PON, POP) organic matter, three nutrient enrichment experiments were conducted with natural microbial communities in shipboard mesocosms, during research cruises in the tropical waters of the southeast Pacific and the northeast Atlantic. Maximum accumulation of POC and PON was observed under high N supply conditions, indicating that primary production was controlled by N availability. The stoichiometry of microbial biomass was unaffected by nutrient N:P supply during exponential growth under nutrient saturation, while it was highly variable under conditions of nutrient limitation and closely correlated to the N:P supply ratio, although PON:POP of accumulated biomass generally exceeded the supply ratio. Microbial N:P composition was constrained by a general lower limit of 5:1. Channelling of assimilated P into DOP appears to be the mechanism responsible for the consistent offset of cellular stoichiometry relative to inorganic nutrient supply and nutrient drawdown, as DOP build-up was observed to intensify under decreasing N:P supply. Low nutrient N:P conditions in coastal upwelling areas overlying O2-deficient waters seem to represent a net source for DOP, which may stimulate growth of diazotrophic phytoplankton. These results demonstrate that microbial nutrient assimilation and partitioning of organic matter between the particulate and the dissolved phase are controlled by the N:P ratio of upwelled nutrients, implying substantial consequences for nutrient cycling and organic matter pools in the course of decreasing nutrient N:P stoichiometry.

Experimental data on photophysiology and growth rates of Pararotalia calcariformata

The eastern Mediterranean is a hotspot of biological invasions. Numerous species of Indo-pacific origin have colonized the Mediterranean in recent times, including tropical symbiont-bearing foraminifera. Among these is the species Pararotalia calcariformata. Unlike other invasive foraminifera, this species has been discovered only two decades ago and is restricted to the eastern Mediterranean coast. Combining ecological, genetic and physiological observations, we attempt to explain the recent invasion of this species in the Mediterranean Sea. Using morphological and genetic data, we confirm the species attribution to P. calcariformata McCulloch 1977 and identify its symbionts as a consortium of diatom species dominated by Minutocellus polymorphus. We document photosynthetic activity of its endosymbionts using Pulse Amplitude Modulated Fluorometry and test the effects of elevated temperatures on growth rates of asexual offspring. The culturing of asexual offspring for 120 days shows a 30-day period of rapid growth followed by a period of slower growth. A subsequent 48-day temperature sensitivity experiment indicates a similar developmental pathway and high growth rate at 28°C, whereas an almost complete inhibition of growth was observed at 20°C and 35°C. This indicates that the offspring of this species may have lower tolerance to cold temperatures than what would be expected for species native to the Mediterranean. We expand this hypothesis by applying a Species Distribution Model (SDM) based on modern occurrences in the Mediterranean using three environmental variables: irradiance, turbidity and yearly minimum temperature. The model reproduces the observed restricted distribution and indicates that the range of the species will drastically expand westwards under future global change scenarios. We conclude that P. calcariformata established a population in the Levant because of the recent warming in the region. In line with observations from other groups of organisms, our results indicate that continued warming of the eastern Mediterranean will facilitate the invasion of more tropical marine taxa into the Mediterranean, disturbing local biodiversity and ecosystem structure.

Impacts of ocean warming and acidification on the larval development of the barnacle Amphibalanus improvisus

The world's oceans are warming and becoming more acidic. Both stressors, singly or in combination, impact marine species, and ensuing effects might be particularly serious for early life stages. To date most studies have focused on ocean acidification (OA) effects in fully marine environments, while little attention has been devoted to more variable coastal ecosystems, such as the Western Baltic Sea. Since natural spatial and temporal variability of environmental conditions such as salinity, temperature or pCO2 impose more complex stresses upon organisms inhabiting these habitats, species can be expected to be more tolerant to OA (or warming) than fully marine taxa. We present data on the variability of salinity, temperature and pH within the Kiel Fjord and on the responses of the barnacle Amphibalanus improvisus from this habitat to simulated warming and OA during its early development. Nauplii and cyprids were exposed to different temperature (12, 20 and 27°C) and pCO2 (nominally 400, 1250 and 3250 µatm) treatments for 8 and 4 weeks, respectively. Survival, larval duration and settlement success were monitored. Warming affected larval responses more strongly than OA. Increased temperatures favored survival and development of nauplii but decreased survival of cyprids. OA had no effect upon survival of nauplii but enhanced their development at low (12°C) and high (27°C) temperatures. In contrast, at the intermediate temperature (20°C), nauplii were not affected even by 3250 µatm pCO2. None of the treatments significantly affected settlement success of cyprids. These experiments show a remarkable tolerance of A. improvisus larvae to 1250 µatm pCO2, the level of OA predicted for the end of the century.

Villefranche sur Mer: multistressors experiment March 2012

The effect of ocean warming and acidification was investigated on a natural plankton assemblage from an oligotrophic area, the bay of Villefranche (NW Mediterranean Sea). The assemblage was sampled in March 2012 and exposed to the following four treatments for 12 days: control ( 360 µatm, 14°C), elevated pCO2 ( 610 µatm, 14°C), elevated temperature ( 410 µatm, 17°C), and elevated pCO2 and temperature ( 690 µatm, 17°C). Nutrients were already depleted at the beginning of the experiment and the concentrations of chlorophyll a (chl a), heterotrophic prokaryotes and viruses decreased, under all treatments, throughout the experiment. There were no statistically significant effects of ocean warming and acidification, whether in isolation or combined, on the concentrations of nutrients, particulate organic matter, chl a and most of the photosynthetic pigments. Furthermore, 13C labelling showed that the carbon transfer rates from 13C-sodium bicarbonate into particulate organic carbon were not affected by seawater warming nor acidification. Rates of gross primary production followed the general decreasing trend of chl a concentrations and were significantly higher under elevated temperature, an effect exacerbated when combined to elevated pCO2 level. In contrast to the other algal groups, the picophytoplankton population (cyanobacteria, mostly Synechococcus) increased throughout the experiment and was more abundant in the warmer treatment though to a lesser extent when combined to high pCO2 level. These results suggest that under nutrient-depleted conditions in the Mediterranean Sea, ocean acidification has a very limited impact on the plankton community and that small species will benefit from warming with a potential decrease of the export and energy transfer to higher trophic levels.