1. WP6 Performance: reproduction and growth PIs: Clemmesen, Catriona, IFM-GEOMAR Piatkowski, Uwe, IFM-GEOMAR Pörtner, Hans, AWI Sommer, Frank, IFM-GEOMAR Widdicombe, Steve, PML EPOCA WP 6

2. Objectives: Identify critical stages in the life cycle (e.g. eggs, larvae) of functionally important marine organisms based on performance measures as indicators of sensitivity to ocean acidification Analyse physiological mechanisms defining performance levels and sensitivity Estimate acclimation capacity (gene expression capacity) for that mechanism as the background of physiological plasticity Quantify impact and tolerance thresholds (tipping points) Assess interaction between ocean acidification (OA) and ocean warming Compare responses and mechanisms in different populations of a species (e.g. in a climate gradient) reflecting potential for evolutionary adaptation (genetic differences) WP6 Performance: reproduction and growth …..as an integrating programme Available funding EPOCA (PhD student equ.): - 1 PhD student AWI - 1 PhD student IFM GEOMAR - 1 PhD student PML Application for complementary national funding underway: e.g. Bioacid, NERC

3. Tasks T1: Sensitivity of critical and early life stages to OA will be assessed through analyses of rates and normality of development, growth, reproductive success, integrity of calcified structures, metabolic and acid-base regulation T2: pH regulation capacity in different body compartments of the animal will be assessed as a potential mechanism shaping sensitivity T3: Regulation capacity will be related to changes in gene expression T4: Impact and tolerance thresholds will be quantified. T5: CO2 effects on tolerance to temperature will be analysed. T6: Responses to CO 2 of different populations, e.g. in a latitudinal cline, will be compared to investigate potential evolutionary adaptation. Work has started at various levels, EPOCA and additional funding. …..as an integrating programme WP6 Performance: reproduction and growth

4. EPOCA WP 6 Experiments will be conducted with the Kiel CO 2 manipulation system and during the 2009 Arctic experiment to determine the impact of high CO 2 on fish and cephalopods eggs, larvae and juveniles. Impact of high CO 2 on Early life Stages of fish and cephalopods Preliminary results (days) C. Clemmesen, U. Piatkowski et al.

5. Homogenate SL Otolith micro- structure RNA/DNA ratio DW, Growth Performance Indicators Histology (PML) IFM-GEOMAR EPOCA WP 6 pH regulation capacity (AWI) RNA/DNA ratio Decrease in condition and protein synthesis capacity with increase in pCO 2 Clemmesen & Franke in prep. Impact of high CO 2 on early life stages of fish and cephalopods C. Clemmesen, U. Piatkowski et al.

6. Impact of high CO 2 on the physiology of key benthic bioturbators. EPOCA WP 6 Amphiura filiformis is an important benthic organism in terms of nutrient cycling and benthic-pelagic coupling. Recent work at PML has shown significant impacts of high CO 2 on this species (Wood et al., 2008 Proc Roy Soc B). Exposure to low pH seawater causes: increase in respiratory rate increased growth increased calcification BUT at the cost of muscle wastage Experiments will be conducted in the PML seawater acidification facility and during the 2009 Arctic experiment to determine the impact of high CO 2 on other key benthic species. A whole organism approach will be adopted to assess both the responses and the costs. S. Widdicombe et al.

7. Hyas araneus, North Sea vs Svalbard n=171-322 extension acceleration EPOCA WP 6 PhD project Kathleen Walther H. Pörtner et al.

8. Antragsskizze BIOACID – Biological Impact of Ocean ACIDificationBonn, 10. Dezember 2007 Goals I.Develop a mechanistic understanding of the combined effects of ocean warming and acidification (OA) on biomineralization/ calcification in high latitude bivalves via whole organism physiology. I.To evaluate the impact of simultaneous temperature rise and OA on “biorecording” properties of bivalve shells, i.e. growth increment formation and biogeochemical parameters Field work - Spitzbergen (AWIPEV station, Ny-Alesund) current status: individuals marked with calcein in 2006, 2007 perspectives: 2008 - recapture specimens from 2006, 2007; mark new animals 2009 - mesocosm experiments (EPOCA) EPOCA WP 6 Greenland smoothcockle (Serripes groenlandicus) NN, O. Heilmayer et al. In situ enclosures modified after Ambrose et. al 2006

9. Long term acclimation via gene expression of pH – regulation mechanisms in fish gills Expression (Real-time PCR) of essential gill transport proteins Na + K + -ATPase Na + HCO 3 - - Cotransporter K. Deigweiher, M. Lucassen, H.O. Pörtner, unpubl. Eelpout (Z. viviparus) 6 weeks EPOCA WP 6

10. Impact of high CO2 and temperature on development and reproduction of mesozooplankton (copepods) To date, the sensitivity of juvenile stages to OA has not been assessed. In the lab we will assess : - growth and development - egg hatching success - respiratory costs and/or gene expression 2 temperatures6 pCO2 levels (~380-4000 ppm) Results will be compared with experiments using constant pCO2 and variable food quality (microalgae grown under different CO2 levels; WP8) In the 2009 Arctic mesocosm we will assess : -copepod egg production, hatching success and naupliar survival EPOCA WP 6 F. Sommer et al.

11. 0 % oxygen limited aerobic scope TcTc TpTp TpTp : Pejus T‘s: Limitation of aerobic performance TcTc : critical T‘s: CO 2 and the concept of oxygen and capacity limited thermal tolerance Shifts in: geographical distributiongeographical distribution Species interactionsSpecies interactions ….food web….food webstructure Anaerobic metabolism Pörtner et al, 2005, Metzger et al. 2007, Pörtner and Knust, Science 2007 100 rate of aerobic perfor- mance 0 Temperature CO 2, hypoxia hypoxia Effects on behaviour,growth, reproduction, ….fitness EPOCA WP 6

12. D6.1: Methodologies used in long term CO2 incubations, presented at meeting (month 9; O, PU) D6.2-3: Internal reports on sensitivities of life stages in individual species (months 12 and 24; R, PP) D6.4: Concept of sensitivity in the life cycles of various groups of marine animals (invertebrates and fish), dissemination as a viewpoint article (month 36; O, PU) D6.5: Conceptual model of mechanisms causing sensitivity and linking organismic responses to ecosystem change (linking to WP 7, 9), dissemination as a viewpoint article (month 36; R, PU) D6.6: Conceptual model of mechanisms and potential for acclimation and adaptation (month 48; R; PU) D6.7: Contribution to mechanism-based model of OA effects at ecosystem level (linking to WP7 and 9) (month 48; R; PU) D6.8: Potentially dangerous thresholds in relevant species, naming uncertainties and probabilities and perspectives on new ecosystem states to PP, PU (linking to WP 13)

14. Addressing CO 2 effects and sensitivities in warming oceans First lines of CO 2 sensitivity (with ecological relevance) likely depend on CO 2 effects on temperature dependent performance in rel. to compensation capacity for extracellular acid-base status. CO 2 effects on temperature dependent performance in rel. to compensation capacity for extracellular acid-base status. This includes disturbance of calcification through extracellular acidification. This includes disturbance of calcification through extracellular acidification. Implications to be considered: seasonal shifts in performance windows seasonal shifts in performance windows climate dependent functional specialization climate dependent functional specialization temperature dependent biogeography temperature dependent biogeography climate dependent growth, fecundity climate dependent growth, fecundity synergistic interactions with factors in addition to temperature (hypoxia, pollutants, …) synergistic interactions with factors in addition to temperature (hypoxia, pollutants, …) Germany United Kingdom

15. Principle considerations: Role of time scales and levels for lethal effects of CO 2 exposure Incipient lethal CO 2 level (long term critical threshold) arbitrary units Mortality independent of exposure time Zone of resistance Mortality dependent on CO 2 level and exposure time Zone of tolerance Upper median lethal CO 2 level (LD 50 ) log exposure time (days, weeks, months, years) → No such complete data set exists Tolerable organism and ecosystem (?) responses Critical level and mechanism unknown? †Acute asphyxiation: squid, fish Pörtner et al., 2005

16. Antragsskizze BIOACID – Biological Impact of Ocean ACIDificationBonn, 10. Dezember 2007 EPOCA WP 6 Impact of high CO 2 on Early life Stages of fish and cephalopods Kiel CO 2 Manipulation system Alarm system Valves and flowmeters for constant pCO 2 air mixtures Control system

17. Impact of high CO2 and temperature on development and reproduction of mesozooplankton Timetable: 200820092010 0-34-67-910- 12 11- 15 16- 18 19- 21 22- 24 25-2728- 30 Growth & development Egg hatching respiration food quality (different microalgae) Arctic mesocosm

18. CO 2 impact on calcification in marine bivalves: a key to understand past, present and future climate records of polar ecosystem Funding: DFG SPP 1158 (HE5753/1-1: 2008 - 2010) Objectives: to develop a mechanistic understanding of the combined effects of temperature rise and ocean acidification (OA) on biomineralization/ calcification in high latitude bivalves to evaluate the impact of simultaneous temperature rise and OA on “biorecording” properties of bivalve shells, i.e. growth increment formation and biogeochemical parameters Greenland smoothcockle (Serripes groenlandicus) circum-arctic distribution subtidal down to 100m 80 - 100 mm within ten years EPOCA WP 6