© 2013Blackwell Publishing 10.1111/gcb.12171doi: lead todifferences version between this andthe ofPleasecite article Version Record. this as been and throughtypesetting, proofreadingwhich may thecopyediting, process, pagination This article undergone has for and accepted been publication fullpeer paper: of Type change; ofOceanAcidification; Calcification; Economic balance. Biochemical impact Keywords author Corresponding UniversiteitAmsterdam de Boelelaan 1085, 6 5 4 3 Spain 2 1 J., 12 title:Running Ocean Acidification Mediterranean red ( coral important economically the on Acidification Ocean of effects Detrimental Articletype : Primary Research Articles Accepted Date 29: Revised Date : 25 Received Date: 25 ScientifiqueCentre de Monaco, AvenueSaint Martin 98000 Monaco, Principality of Monaco Institució CatalanaRecerca de i Estudis Avançats (ICREA), Barcelona, Spain ICTA InstitutdeCiències del Mar, ICM California State University Northridge. 18111 Nordhoff 91330 st. Northridge,CA, USA Department of Earth Sciences Bramanti L., Bramanti

Accepted Article2 Lopez -

UAB, - Sanz A., Sanz UAB Campus Cn Cerdanyola s/n delVallés 08193 Barcel : Mediterranean red coral; coral; red Mediterranean : 2

Movilla J., J., Movilla Primary ResearchPrimaryArticle - - - Jan Jan Jan 2 Martinez - - - 2013 2013 2013

: Lorenzo Bramanti8184708903e Tel:+1

3 Guron M., Guron - -

Ltd Quintana A., Earth and Climate, - CSIC, Passeig Marítim de laBarceloneta 37 Corallium rubrum

Corallium rubrumCorallium

eaoi efcs f ca Aiiiain Climate Acidification; Ocean of effects Metabolic 2

1081HV Amsterdam, The Netherlands Calvo E., Calvo

24 Pelejero C.,

Faculty Earthof and Lif 5 Gori A., Gori )

36 Ziveri, P., 2 ona, ona, Dominguez - Spain. mail: mail: e Sciences,e

review but has not review 3 - Rossi S. 49, 08003, Barcelona,

[email protected] - Carrió C., C., Carrió

VU

2

Grinyó

© 2013 Blackwell Publishing Ltd o surface averaged global the than larger 2011), Goyet Acceptedpre the since units 0.14 to up of decrease pH a estimated study CO atmospheric increasing 2009) Goyet & Touratier 2010; al. CO anthropogenic of penetration fast and concentration (50 waters its of time turnover Mediterranean the of case the In locally. varies impact its scale, global a at acts CO considered the on depending futu and already pre the since oceans world’s the hasof waters surface the affected units pH 0.1 of reduction average An 2009). al. et (Doney years coming the 1. Introduction Articlespecies. coral endangered and valuable this on effects acidification ocean detrimental of evidence time, first contr to thefor show, results Our relative colonies. the metabolism of the on decrease pH conditionsof effect possible a suggests acidified in maintained samples among found variability lower the However, composition. acid fatty and protein lipid, carbohydrate, total regarding treatments between organic under total higher while significantly hand, was other matter the On aberrant conditions. acidified with the experiment, under the only of observed being end shapes the at treatments both between treatment differed control clearly the morphology to comparison in rate growth skeletal the in decrease significant a caused conditions pH lower to Exposure periodically. measured were composition acids morpholspicule rate, pH 7.81, and (8.10 levels pH different two at aquaria in days 314 for maintained term of coloniesexperiments. Severalaquaria speciesfrom this on pH low of effects decades. next the organismover this for bleakfuture a depicts species, Mg its solubilityof elevated the ( coral red the on acidification ocean Mediterranean the of them, Among organisms.calcifying effectson emphasis strong with environment, potential marine the assess to research urgent prompted global has the century in units pH 0.4 to 0.3 of decrease predicted mean The Abstract

Ocean acidification Ocean re projections predict a decrease by 0.3 to 0.4 pH units by the end of the century, the of end the by units pH 0.4 to 0.3 by decrease a predict projections re

Corallium rubrum Corallium

ogy, major biochemical constituents (protein, carbohydrates and lipids) and fatty and lipids) carbohydratesand biochemicalconstituents(protein, ogy,major )

2 is expected to be particularly susceptible to acidification effects, due to due effects, acidification to susceptibleparticularly be to expected is (OA) is considered a major threat to the marine environment in in environment marine the to threat major a considered is (OA)

- Ymz t l 20; av e a. 01 Zvr 21) A recent A 2012). Ziveri 2011, al. et Calvo 2008; al. et (Ylmaz calcite skeleton. This, together with the large overexploitation of thisoverexploitation of large the with together This,skeleton. calcite

2

- emission scenarios (e.g. Joos et al. 2011). Although OA Although 2011). al. et Joos (e.g. scenarios emission

100 years; Bethoux et al. 2005) and the very high high very the and 2005) al. et Bethoux years; 100

o p cniin, o infcn dfeecs ee detected were differences significant no conditions, pH low makes it one of the world’s most most world’s the of one it makes 2

cean pH decrease of of decrease pH cean (Schneider et al. 2007; Schneider et Schneider 2007; al. et (Schneider - ufc oen y h ed f the of end the by ocean surface industrial era (Orr et al. 2005) al. et (Orr era industrial In this study, we evaluated the evaluated study,we this In - industrial era (Touratier & (Touratier era industrial Smlry te spicule the Similarly, . sensitive regions to to regions sensitive C. rubrum C. T ). Calcification ). 0.1 pH units units pH 0.1 Sea, the fast fast the Sea,

were long were ols, - © 2013 Blackwell Publishing Ltd Acceptedpathwaysbe can more in Hoegh 2009; Peltzer & (Brewer stresses environmental other counteract to or growth tissue reproduction, availabilit its reducing process, this for energydemand the determining in factors key are decrease pH seawater magnitudeof the and changeof rate the Itlikelythat 2012). is al, et McCulloch 2012; GaoLi& 2012; al. et 20 al. et (Wood organism the for cost metabolic high a and energy of amount significant a requires process physiological active this yet 2011), al, [CO and pH raise (Al site calcifying to capacity the have species some that evidence is of 2009). There Holcomb & site Cohen 2011; the al. et at (Allemand understood poorly pH still is the calcification controlling mechanisms physiological/biochemical critical on O of impact the and complex highly is corals in process calcification the therein).However, Articlereferences and 2012 Connolly & Chan 2010; al. et Kroeker (e.g., formation skeleton the and reduc the to associated ion carbonate of concentration decreased the that indicate experiments most communities, 2008; resvariety wide of a shown haveorganisms instancesLejeusne(Crain synergistically etal. et 2008; al,2010). al,2010;Coll a organisms marine impact can which 2011) al. et Madron de Durrieu 2011; al. et Calvo (e.g. stressors environmental important other by characterized also is Sea Mediterranean The 2005). al. et Orr (e.g. normally recognised

Ries et al. 2009; Wicks & Roberts 2012; Suggett et al. 2012). In the case of coral reef coral of case the In 2012). al. et Suggett 2012; Roberts & Wicks 2009; al. et Ries marine on levels pH projected the of impact potential the examining studies Several

- udeg t l 20) Teeoe suis iig o dniy hc metabolic which identify to aiming studies Therefore, 2007). al. et Guldberg - Horani et al. 2003; Cohen & McConnaughey 2003; Ries 2011; Trotter et Trotter 2011; Ries 2003; McConnaughey & Cohen 2003; al. et Horani tion in pH has a detrimental effect, hindering the calcification process calcification the hindering effect, detrimental a has pH in tion fluenced such may an play by OA effect. rolea tounderstand helpful

t udrae te atvte sc a locomotion, as such activities other undertake to y ponses (e.g. Doney et al. 2009; al. et(e.g. Doney ponses d csses n ifrn wy, n many in ways, different in ecosystems nd 08; Cohen & Holcomb 2009; Edmunds 2009; Holcomb & Cohen 08; Guinotte & Fabry,& Guinotte

3 2 - a the at ] A

© 2013 Blackwell Publishing Ltd Acceptedthese new environments. among and conditions pH of decrease the to susceptible more be is it Thus, 2005). al. Mg that et conceivable Orr 2008; al. et (Andersson carbo latitude increasing to with respect decreases with state saturation seawater the and 1974), Mckenzie, Mg of solubility CaCO of stocks temporary as act may they that suggested been has it although 1993), (Allemand The 1993). al. et (Grillo fusion their of product the not is it but 2008) al. et Vielzeuf calcite, rich (Mg sclerites free the than composition mineralogical same the has skeleton axial The μm). thick a lies, mesoglea the which under ectoderm, the by formed is coenenchyma the of surface external tissue The (coenenchyma). living with coated sclerites and skeleton axial an of composed is coral Red 2011). Articlespeci flagship or patrimonial a as considered be 2004). al. et can coral red (Santangelo value, aesthetic landscape and overexploited importance cultural rarity, its considered to due Moreover, now is it and times ancient since reasons, these For 2010). al. et (Tsounis industry jewellery the in material raw as used skeleton durable red bright its to due corals precious valuable most the of one as considered is It 2008). al. et Rossi 2010; al. et (Costantini depth m 600 and 10 betwe found be can where rockyit neighbouring Atlantic shores, its andMediterraneanSea long a is It Gold”. Red “Mediterranean t of One 2009). al. et (Cooley species interest commercial high of harvest the in reduction the with A ol as hv cer einl socio regional clear have also could OA

an ucin f h slrts s o rvd mcaia poeto aant abrasion against protection mechanical provide to is sclerites the of function main 3 , readily available for the formation of the axial skeleton (Vielzeuf et al., 2008). The The 2008). al., et (Vielzeuf skeleton axial the of formation the for available readily , hese key species is the red coral, red the is species key hese - rich calcite minerals is greater than that of aragonite or calcite (Plummer & (Plummer calcite or aragonite of that than greater is minerals calcite rich - rich calcite, temperate calcifying organisms such as as such organisms calcifying temperate calcite, rich

acellular layer of collagen containing small sclerites (30 sclerites small containing collagen of layer acellular - Corallium rubrum Corallium ie, slow lived, - cnmc aiiain sc a toe related those as such ramifications economic es of the Mediterranean Sea (Bramanti et al. et (Bramanti Sea Mediterranean the of es - growing gorgonian endemic to the the to endemic gorgonian growing

(Linnaeus 1758), the so the 1758), (Linnaeus C. rubrum C. the first to be affected by affected be to first the

has been harvested harvested been has C. rubrum C. nate minerals minerals nate

- should should called - 50 en - © 2013 Blackwell Publishing Ltd were Colonies (Spain). Barcelona in CSIC) (ICM Mar del Ciències de Institut the ± to 0.5ºC) (12.5 Accepted temperature constant at containers seawater large in transported were specimens Sea, 003°19 Mediterranean 42°19’N; NW (Spain, Creus de Cap of Area Protected Marine the from depth collection 2.1 Specimen experimentaland setup methods2. Materials and How OA will affect redc the harvestable stocksof (5) energy?; store to capability organism’s the with related responses physiological different to lead OA Could (4) acids?; fatty and lipids proteins, carbohydrates, matter, organic total of OA does How (3) sclerites?; of morphology CaCO the affect pH Articleeconomicecologicalconsequences and impacts precious this ofOA on rubrum of response the on information valuable provide results The evaluated. were conditions differen under reared colonies the of composition biochemical and morphology sclerite rate, calcification in changes potential The century. the of end the by projected conditions pH inve we topic, this information of on light shed To lack harvested. be to potential with colonies these a in effects OA future regarding is there However, 2009). al. et Bramanti 2007; al. et (Santangelo be have models dynamic 2011), 2010, GFCM 2010; al. et Bussoletti 2009; Roberts & (Brukner octocoral precious this of Owing to the demand for management and conservation plans for a sustainableaharvestingfor plans conservation managementand for demand the Owingto oois of Colonies in lowering a Does (1) questions: following the answering at aimed we study, this In

to low pH conditions that will increase our understanding and capacity to forecast the the forecast to capacityand understanding our increase will that conditions pH low to ’E stigated the effect of OA on OA of effect the stigated ) in November 2010 when in situ temperature was 13 ºC. The collected The ºC. 13 was temperature situ in when 2010 November in ) . rubrum C. 3

eoiin in deposition

ee aeul rmvd rm ok sbtae a 35 at substrates rocky from removed carefully were en developed, projecting demographic trends over time time over trends demographic projecting developed, en . rubrum C.

affect the metabolic balance of col of balance metabolic the affect C. rubrum C.

?; (2) What is the effect of OA on the the on OA of effect the is What (2) ?; oral?

by simulating in aquaria the future future the aquaria in simulating by

species. species. onies in terms terms in onies

- 0 m 40 t pH pH t C. C. © 2013 Blackwell Publishing Ltd (HYDOR pump a with mixed was aquarium each in Water maintained. were colonies where aquaria experimental the to hour) per L (12 continuously flowed tanks large the from Water CO the using treatments, both in seawater in system carbonate the of parameters of rest the calculate to thanused were pH and TA precision electrodes. better provides which 1993), Byrne & (Clayton spectrophotometry using 1987; Fraga, & (Perez titration potentiometric by (TA) alkalinity total analyse to experiment) the of rest the for bimonthly and months 3 first the during month a (once periodically taken were water of volumes small addition, In 2007). (SOP6 al. et procedures Dickson standard following buffer, Tris a with basis daily a on calibrated were electrodes glass The USA). Inc., Topac R316, (Consort controller pH a to connected Sea CO or purity) (99.9% al. et CO ppm ~800 and CO ppm ~380 of atmosphere an with equilibrium in seawater Mediterranean the respectively, simulating, scale) (total units pH ~7.81 and ~8.10 to up L 150 of tanks large pH Seawater S2. Fig. in shown is up set 2 into subdivided further were experimental The below). (see pH low and pH control Aquaria treatment), per replicates (3 treatments each). litres (30 aquaria 6 among distributed of colonies 48 acclimation, of month ( de feeding, For darkness. complete in and temperature constant at maintained were tanks the in colonies small, very are variation temperature seasonal the coral where 2013), red the As respectively). e (Tsounis depth meters 100 and 37.6, 80 between dwells populations harvested commercially and (12ºC conditions salinity and temperature maintai Accepted Article a per weight dry of mg (~55 daily supplied were ™) Nutrition Ocean water pH was continuously monitored by glass electrodes (LL Ecotrode plus Ecotrode (LL electrodes glass by monitored continuously was pH water

08. n re t ahee h dsrd H ees saae ws ube wt CO with bubbled was seawater levels, pH desired the achieve to order In 2008). ned in a 100 L acclimation tank with 50 μm filtered running natural seawater atseawater running filterednatural μm 50 with tank L acclimation 100 a in ned 2

( future levels predicted for year 2100 following A2 IPCC SRES; PlattnerSRES; IPCC A2 followingyear 2100 for predicted levels future 2 - free air (using a filter filled with soda lime absorber, Sigma Aldrich). Aldrich). Sigma absorber, lime soda with filled filter a (using air free

. rubrum C. was gradually adjusted (0.03 units per day) in two in day) per units (0.03 adjusted gradually was 2 calc sof calc

Fg S) ee eetd n randomly and selected were S1) (Fig. tware tware (Robbins et al. 2010) al. et (Robbins Perez et al. 2000) and pH pH and 2000) al. et Perez uru) Atr 1 After quarium). 2 -

(current levels) (current frozen

- (Table S1). (Table

Metrohm) Cyclops Cyclops in situ in of a t al, t 2

© 2013 Blackwell Publishing Ltd water.distilled with Acceptedwashed andmatter organic removethe to hypochloritedays duringtwo sodium in dipped was treatment) per (3 aquarium each from colony one this, For technique. (1998) al.’s et Bucher gramper perwas day.usedfor analyses. weight ofinitial Thisnormalization statistical CaCO of mgas expressed is (G)growth rate the and mass initial their to normalized cm g 2.66 of value density specific the using weight dry to transformed was glue) and holder coral the minus weight coral (total corals the YSI an using monitored constantly were seawater of measurements, the During SFACT). AB204 Toledo (Mettler balance (Jokiel weight buoyant of measurements 2.2 Articlemicrodensityand porosity. specific estimate to aquarium each from selected randomly was colony extra one experiment, the of end the at Furthermore, undertaken. were analyses biochemical and morphological at kept and removed were aquarium) experimental each from (1 colonies random 6 and measured was colonies ofall the weight event, buoyant each the During sampling 3). November 2011(Time controlled during room, ensuring (~12°C) whole constant experiment. values the pH The exchange. gas h l 1500 4.5W, Koralia; Skeletal measurements Microdensity and porosity of the of porosity and Microdensity the of growth in Changes rubrum C. - 0C sbeunl freeze subsequently 80ºC,

oois ee ape qu sampled were colonies Buoyant weight and dry weight of each sample were recorded before and recordedbefore wereeach sampleof weight dry andBuoyant weight - manipulative experimental set experimental manipulative - 1 ) and a plastic a and )

. rubrum C.

wrap was used to reduce evaporation and surface and evaporation reduce to used was wrap skeleton of skeleton et al et - dr . e ad hn trd rzn at frozen stored then and ied 1978; Davies 1989), using a 0.1 mg resolution mg 0.1 a using 1989), Davies 1978; arterly, from December 2010 (Time 0) to to 0) (Time 2010 December from arterly, - 3

( oois ( colonies e blw. h ices i wih was weight in increase The below). see C. rubrum C. - - 30M probe. The net buoyant weight of of weight buoyant net The probe. 30M up was installed inside a temperature a inside installed was up n

were evaluated were = 30) were assessed from from assessed were 30) = temperature and salinity salinity and temperature

following the the following

- 0C until 20ºC 3

increase

- air - © 2013 Blackwell Publishing Ltd ( colony each from coenenchyma biochemical2.4 Major constituents measure of compactness (Turon & Becerro, 1992). arepresents perimeter,same the with circle a of area the and sclerite the of area the between ratio the f the sclerite, each perimeter, of shape Area, overall the 2004): describe to al. order In et height. maximum (Abramoff and width J maximum Image software graphic the of means by recorded anomalous an with Sclerites X. 1300 labelled of were morphology magnification a using measured were 15) = II Type 20, = I (Type sclerites 35 colony, each For 1975). Grasshoff 3) & Carpine S3; (Fig types morphological time and 0 time for treatment per replicates S HITACHI (SEM) Microscope Electron Å) 200 (< palladium gold co and silver colloidal with stub aluminium an to attached cover glass a on deposited was sample the of portion small Amounting. during drying faster for ethanol absolute with were sclerites and dissolved was Aft disaggregated. matter organic until solution hypochlorite sodium in dipped wasbranches, the of height midapproximately at taken colony, each of mmlength 5 about of sample 2.3 Sclerite morphology for equations each describedBucher sample using the al. et(1998). in cm cases, the (105 wax paraffin molten in inclusion the after Acceptedas defined latter, The circularity. and width/height area/perimeter, calculated: were ratios ollowing Article – 3 . Total enclosed volume, skeleton matrix volume and bulk density were also calculated also were density bulk and volume matrix skeleton volume, enclosed Total . rai mte (M o piay rnhs a dtrie i 10 in determined was branches primary of (OM) matter Organic Following methodology the described Lewis & in buoyant weight was measured in distilled water at 20ºC with specific density waswaterat withspecific measured weight indistilled 20ºC buoyant 1.00 r eietto, h smls ee isd ih itle wtr n subsequently and water distilled with rinsed were samples the sedimentation, er

. s brat Fg S) Fr ah ceie h floig esrs were measures following the sclerite each For S3). (Fig. aberrant as ceie opooia osrain wr promd ih Scanning a with performed were observations morphological Sclerite

n = 24). To obtain dry weight, samples were heated at 80ºC 80ºC at heated were samples weight, dry obtain To 24). =

- 50 wrig t . K. ttl f 2 oois (3 colonies 12 of total A KV. 5.0 at working N 3500

–

110ºC) to form a water a form to 110ºC) ee xmnd n ctgrzd n w main two in categorized and examined were

von Wallis (1991) andGori

vered with a thin layer of layer thin a with vered - tight barrier. In both In barrier. tight - 12 mg of dry dry of mg 12 et al.et (2012a),

g a © 2013 Blackwell Publishing Ltd f of esters methyl The water. saturated NaCl of ml 4 with quenched Accepted was fraction acids fatty was reaction The h. 1 for 90ºC The at heated and MeOH/BF3 20% of solution a using methylated (98:2). acid ether:acetic diethyl of ml 8.5 with recovered chloroform:2 of ml 3 with eluted was fraction first The of ml 4 with activated previously was that 2007) Werne & (Russell column glass aminopropyl mg 500 a through eluted and chloroform of ml 0.5 in redissolved was sample constant a at Ruiz concentrator to according extraction vacuum phase solid by centrifugal fractionated and temperature a in dryness near to taken was extract extracted (2 mixture standard internal an with (dichloromethane DCM:MeOH 3:1 in homogenized were sample each OM. of These method mg per constituent of µg in presented are results the All standard. as cholesterol using (1973), Blackstock & Article Barnes of method the to according colorimetrically quantified homogenized were sample each from weight dry coenenchyma of mg 12 approximately Finally, standard. as albumin Lowry of method the using colorimetrically quantified was content protein s each from weight coenenchymadry Dubois of method the to according colorimetrically quantified was content carbohydrates total and water, distilled double of 3ml the f coenenchyma of dry of weight mg 8 dry Approximately sample. initial the to respect with percentage as expressed are Results 1995). McClintock & (Slattery weight ash and dry between difference the as calculated then was hours 5 for ashed then were samples same The h. 48 for F or fatty acid proportions estimation, approximately 10 mg of dry coenenchyma from from coenenchyma dry of mg approximately10 estimation, proportions acid fatty or using microwave assisted extraction for 5 min at 70ºC. After centrifugation, the the centrifugation, After 70ºC. at min 5 for extraction assisted microwave using ologies havealreadyologies applied been with in 3 ml of chloroform of ml 3 in

et al. et ample were homogenizedin wereample - cyddcni ai ad 5β and acid octyldodecanoic (1956), with glucose as standard. Another 8 mg of of mg 8 Another standard. as glucose with (1956), - methanol (2:1) and total lipid content was content lipid total and (2:1) methanol rom each sample were homogenized in homogenized were sample each rom C. rubrum C. - rpnl 21 ad the and (2:1) propanol at 450ºC and weighted again. OM again. weighted and 450ºC at

(Rossi &(Rossi 2007). Tsounis,

2 ml of 1N NaOH and total and NaOH1N of ml 2 atty acids were recovered were acids atty - cholanic acid) and and acid) cholanic - ehnl, spiked methanol), et al. et et al. et

(1951), with (1951), (2004). Th (2004).

fatty acids acids fatty n - hexane.

e © 2013 Blackwell Publishing Ltd one A (SE). error standard ± mean as expressed are data Acceptedrate Calcification weighted. were treatment) per 15 aquarium, t A treatment. within nested factor random a as considered was Aquarium aquaria. and conditions) pH control to exposure and conditions pH low to exposure (i.e., treatments analyses2.6 Statistical 2003). has Thismethodology ingorgonians alreadyapplied (Gori been2012b). etal. concentrati with those only PUFA), Acids, Fatty Unsaturated Poly and MUFA Acids, Fatty Unsaturated converting them and software, 4.1 Chromquest the using areas peak the integrating by quantified were acids C4 Mix Supelco® compounds, FAME (29 acids fatty standard of those with esters Methyl respectively. Article min 4ºC at 300ºC to min to 50ºC from increase to programmed min ml phase 30 at μm gas carrier 0.25 a as and used was diameterHydrogen thickness). internal mm 0.25 length, m (60 column Agilent DB5MS Agilent an with a with equipped performed GC 7820A Technologies was analysis chromatography gas and isooctane of μl 80 in at stored was sample c the of removal After water. residual remove Na with filled column glass a through eluted and chloroform of ml 1.5 in redissolved of ml 3 with twiceby extracting - 1 , from 188ºC to 299ºC at 20ºC min 20ºC at 299ºC to 188ºC from , A two A Mono SAFA; Acids, Fatty Saturated (total classes into grouping acid fatty For n hge ta 1 o te oa fty cd wr cniee (Daalsgaard considered were acids fatty total the of 1% than higher ons - way nested ANOVA was used to examine whether calcification rate varied between varied rate calcification whether examine to used was ANOVA nested way - into concentrationsareainto from the way ANOVA was used to examine differences between both treatments (Control treatments both between differences examine to used was ANOVA way - - 1 20ºC until analysis by gas chromatography. Samples were redissolved were Samples chromatography. gas by analysis until 20ºC

Ijco ad eetr eprtrs ee e a 30C n 320ºC, and 300ºC at set were temperatures detector and Injector . of fatty acids were identified by comparing their retention times retention their comparing by identified were acids fatty of

n - hexane. The combined extracts were taken to neardryness, were to takenextracts combined The hexane.

160ºC at 20ºC min 20ºC at 160ºC - 1 , from 299ºC to 235ºC at 2ºC min 2ºC at 235ºC to 299ºC from , flame ionization detector, a splitless injector and a and injector splitless a detector, ionization flame lrfr udr irgn a, h extracted the gas, nitrogen under hloroform vs

concentration of theinternalstandards. - 1 , from 160ºC to 188ºC at 0.5ºC 0.5ºC at 188ºC to 160ºC from , - 1 . The oven temperature was was temperature oven The . otal of 30 colonies (5 per (5 colonies 30 of otal - 1

and from 235ºC 235ºC from and - 2) Fatty C24).

2 SO t al. et 4

to

© 2013 Blackwell Publishing Ltd Acceptedaquaria replicates of the same treatment anyin analysis. Fig. 1). The survivorship in ea calcification whichrates, were found beto 59% lower compared to control conditions (Table 1and rubrum which wasit subjected. structural material composition of this gorgonian was affectednot by the level of acidification to (ANOVA, F respectively.6), No significant differences between treatments were observed, neither microdensityin Microdensity andporosity of 3.1 3. Results using the R software platform (R development Tea the performed were using analyses statistical performed other the was all while 2005), analysis (Anderson software Multivariate PERMANOVA calculated. was Mean) Article / Deviation (Standard the pH, low to response the to associatedvariability the of measure a have to orderIn (SD). deviationstandard ± mean as expressedare biochemicaldata All only.interaction their differenc significant honest Tukey’s differences, significant showed ANOVA When points). sampling (4 time and pH) low and (control treatment were case this in Factors results. biochemical analyse to used was ANOVA way treatment within nested time), factor (random aquaria and pH) final low and (control and (initial time were Factors treatments. between differences significant no of hypothesis null the test employedto was 2001) (PERMANOVA;Andersonvariance of analysis distance A standardizeddata. the on based built was matrix distance Euclidean a and (MAD), deviation absolute median their to respect with standardized were results poro and microdensity in pH) low and Skeletal measurements , a significant decrease was observed at the end the of experiment in their averaged 1, 5 1,

= 5.25,p = nor0.08) on porosity (ANOVA, F Regarding effect the of the lower pH tre

e test (HSD) was used to attribute differences between specific factors or factors specific between differences attribute to used was (HSD) test e C. rubrum ch treatment was 100% and no tank effect was detected between

were 2.66 0.06± g cm iy f h Mg the of sity m 2012).

- ih act seeo. ceie morphology Sclerites skeleton. calcite rich

1, 5 1, - 3

atment on skeletalthe growth of

= 0.42,p =0.55),

and 25.7 ± and 25.7± 5.3 % (mean SD; n ± = treatment). Finally, a univariate two univariate a Finally, treatment). - based permutational multivariatpermutational based

Coefficient of VariationCoefficientof indicating that the C. e - © 2013 Blackwell Publishing Ltd 4). (Fig 3 and 2 times in values variability of coefficient higher showed conditions control under reared samples case, previous the in As S4). Fig PUFA; and MUFA (SAFA, proportions acid fatty the in Two amongdifferenttimes levels4;(Figacid fatty total the Variation (CV) was higher thein control F 0.05; > P (Two experimentwhole the duringcontent lipid carbohydrate or protein, significant differences were found in the lower pH conditions throughout the experiment. gr of OM the While conditions. control in that than higher remained treatment pH low the samplescontentofmean inOM and the T3, while T2 in differencesbetween andT1, treatments T0 in (Table “Time”factor the and “Treatment” 3.3 acidified treatment. classif shapes anomalous of presenceFurthermore,the treatment. control the with compared significantlydifferent was conditions pH low under grown colonies the of sclerites of shape overall the days) 314 (after 3 Time at while differen significant no were there experiment) the of beginning the (at 0 Time at that showed test hoc post The scleriteshape. overall the on pH low of effect an of hypothesis both for Treatment factor 3.2 Sclerites morphology

Accepted no 2), Fig. and 3 (Table T3 to T0 from changed significantly treatment control the in own Article

Major biochemicalMajor constituents Regarding fatty nosignificant acids, differences foun were treatments between differences no balance, metabolic the Regarding factor the between interaction significant a showed content OM the of analysis The the and Time factor the between interaction significant a showed results PERMANOVA 3,16

= 1.22, p > 0.05; F 0.05; > p 1.22, =

types of sclerites (type I and II, Fig. S3, Table 2). These results confirm theconfirm results These 2). Table S3, Fig. II, and I (type sclerites of types ied as aberrant type (Fig. S3) was detected only in colonies reared in the in reared colonies in only detected was S3) (Fig. type aberrant as ied

3,16

= 0.009, p > 0.05, respectively). Furthermore, the Coefficient ofCoefficient the Furthermore,respectively). 0.05, > p 0.009, =

samples 3 and Fig. 2). Post hoc comparisons (Table 3) indicated noindicated 3) (Tablecomparisons hoc Post 2). Fig. and 3

(Fig 3).

were found

- way ANOVA,F way

between treatments, neither thein - ces between both treatments both between ces way ANOVA, F ANOVA, way 3,16

= 0.49, P > P 0.49, =

3,16 C. rubrum C.

i total in d 0.05), nor0.05),

= 0.093, =

© 2013 Blackwell Publishing Ltd of appearance and rate growth skeletal Accepted disruption and corrosion errata Schizoporella (Rodolfo 7.3 as low as levels observedin of skeleton exposed the CO volcanic by influenced gradient pH natural cora temperate which in date to experiment only the addition, In caespitosa. C. of rates calcification the on (7.88) pH seawater decreased of effects no showed and patagonica rat calcification in reduction 35% a found (2012) al et Movilla while 7.4, of pH a at for dissolution skeleton total reported (2007) Tchernov & Fine species. therein) references and 2011 (Ries, the process in calcification complexity intracellular some the to controlling pointing mechanisms 2013), al, et Comeau 2010; al. et Ries 2010; al. et Articlehowever, tocalci are able of case the in reduction of 70% to up reaching 2006), al. et Kleypas 2008; Fabry, and (Guinotte conditions calcificati the in decline the corals,scleractinianIntropical case of the calcifying species. benthic other of OA responseto the examining studies experimental previous with consistent is it and formation skeletal on confir reduction This 8.10). (pH treatment control the in maintained those with compared 59% of rate growth skeletal in decrease a suffered 7.81) (pH in structural4.1 Changes the 4. Discussion After 314 days of exposure, colonies of of colonies exposure, of days 314 After . Similarly, a wide range of responses has been observed in Mediterranean coral coral Mediterranean in observed been has responses of range wide a Similarly, . Balanophyllia europaea Balanophyllia

oie rus Porites ldcr caespitosa Cladocora

transplanted to the same naturally acidified site showed skeletal skeletal showed site acidified naturally same the to transplanted fy even under low saturation state conditions (Jury et al. 2010; Krief state conditionsal. (Jury 2010; et fy saturation even low under in calcification at pH 7.76 (Lombardi et al. 2011a). However, the the However, 2011a). al. et (Lombardi 7.76 pH at calcification in . caespitosa C.

t H .0 Melenr Emns 20) Sm species, Some 2008). Edmunds, & (Muehllehner 7.80 pH at

n ae ags rm 0 t 6% t obe elevated double at 60% to 10% from ranges rate on features ofC.rubrum

- eap e a. 01. eadn Mdtraen bryozoans, Mediterranean Regarding 2011). al. et Metalpa , which skeleton was completely covered by tissue, at pH pH at bytissue, covered completelywas skeleton which ,

t p o 78, n Rodolfo and 7.83, of pH a at Myriapora truncata Myriapora

at pH 7.5 environments, while no effects were were effects no while environments, 7.5 pH at C. rubrum C. 2

vents, showed evidences of dissolution on dissolution of evidences showed vents,

ms the expected detrimental effects detrimental expected the ms

reared under lower pH conditions pH lower under reared

were not affected until a pH of of pH a until affected not were s ee rnpatd o a to transplanted were ls - Metalpa et al (2010a) (2010a) al et Metalpa Oculina patagonica Oculina es for O. for es p CO 2

© 2013 Blackwell Publishing Ltd Acceptedcapacityas well biomechanical as theskeleton properties. affect will OA overall their our defining Therefore, 1991). Wallis factor Von & (Lewis determinant structure a being movements, these of extent the limit tissue the in sclerites of abundance and morphology The 1987). Lewis, & (Jeyasuria position forwa bend 2008). to colonies the allow that properties elastic with al. 1993) et al. et (Grillo proteinaceous (Vielzeuf typically is skeleton the recrystallization Alcyonacea, order the in and Furthermore, transport, dissolution, of mechanisms CaCO as act may they that suggested been has it although abrasion, against protection mechanical the Acc high to exposed larvae urchin sea in previously observed been has skeletogenesis abnormal Sclerites conditions. acidified in only wit sclerites, Articleto OAintheMediterranean. organisms susceptible most the of one as considered be could species this therefore, and, two la this of solubility the mentioned, previously As 2008). Mg of entirely consists structure skeletal coral red the 2006), al. et Smith 2003; McConnaughey, & (Cohen aragonite or calcite of made are skeletons whose seawater. pH Rodolfo 2009; organic Rodolfo 2011b; al. et (Lombardi 7.43 ording to Allemand (1993), the main function of these sclerites in sclerites these of function main the (1993), Allemand to ording u rsls lo hwd n fet f oe p o te opooy f microscopic of morphology the on pH lower of effect an showed also results Our

tissue enveloping the skeleton could play a key role in these organisms (Ries et al, et (Ries organisms these in role key a play could skeleton the enveloping tissue 3

eprr sok raiy vial fr h ail kltn format skeleton axial the for available readily stocks temporary h an h It should be noted that, unlike scleractinian corals or bryozoans bryozoans or corals scleractinian unlike that, noted be should It - Metalpa et al, 2011), providing protection against the corrosiveness of lower lower of corrosiveness the against protection providing 2011), al, et Metalpa . rubrum C. overall shape different between treatments and anomalous forms observed forms anomalous and treatments between different shape overall

n ohr ognas n opoiig h CaCO the compromising in gorgonians other and

- Metalpa et al, 2010b), al, et Metalpa p CO 2

conditions (Kurihara & Shirayama, 2004). 2004). Shirayama, & (Kurihara conditions

results suggest that, in the long term, long the in that, suggest results tter form is much higher than the first the than higher much is formtter

suggesting that the presence of of presence the that suggesting - rich calcite (Vielzeuf et al. et (Vielzeuf calcite rich rd and back to an upright an to back and rd C. rubrum C. as o through ion

is to ensure to is 3

S. errata S. stocking , © 2013 Blackwell Publishing Ltd Rodolfo 2009; al. et Ries 2010; Acceptedo effects the modulate may andseawater to skeleton the of exposure direct the prevents layer tissue a of presence The 1991). Wallis von collagen of material & (Alderslade gel mesoglea of layer thick a by protected 4.2 coral extent which to colonies inhabiting ranges be shallower will threatened. establish and stressors multiple between interactions possible of existence the assess to temperature and OA manipulating studies future perform to essential T 2008). al, et Torrents 2009; 2001, al. et Garrabou 2005; al. et (Bramanti recruits and colonies the of mortality total or partial in result can temperature high to exposure term activity polyp the year. the throughout depress constant and rates respiration and metabolic increased induce temperatures relatively high Nevertheless, remain to tend and Article marked as not are changes be (i.e. potential harvest with coral red water shallow populations on focus particular affecta has study present the 2012a), al. et (Santangelo populations well could acidification ocean and warming of effects Gar et (Coma events with mass mortality al.2009; associated periods stratification longer into translate to shown been has temperature seawater increased Sea, Mediterranean the In temperature. seawater global in increase an causing responsi is pCO2 atmospheric increased While OA. with synergy in or independently acting populations, coral red future affect may pressures environmental other OA, to addition in However, coral. red the as such value commercial high with anthozoan Metabolic response of red coral to a lowering in pH Metabolic response ofredin coral toalowering This study focused on evaluating the effect of OA on the growth of a Medit a of growth the on OA of effect the evaluating on focused study This ognas kltn osss f nenl omtos cnrl xs n sclerites) and axis (central formations internal of consists skeleton Gorgonians

in red coral and other gorgonians (Previati et al. 2010). In addition, long addition, In 2010). al. et (Previati gorgonians other and coral red in - iefbis admyoine n yrtdplmrmti Lws & (Lewis matrix polymer hydrated a in oriented randomly fibrils like

low 60 meters depth, Rossi et al. 2008), where temperature temperature where 2008), al. et Rossi depth, meters 60 low - Metalpa et al. 2011). However, in our experiment, although experiment, our in However, 2011). al. et Metalpa f OA in coral and other organisms (Hoffman et al. et (Hoffman organisms other and coral in OA f rabou et al. 2009). Although the synergistic Although synergistic raboual. 2009).the et

arcu 20) a composite a 2008), Fabricius ble for OA, it is also is it OA, for ble hus, it is it hus, erranean - © 2013 Blackwell Publishing Ltd observed that to comparable is aquaria Accepted of the biochemical indicating composition that 3), conditions (Fig. natural con environment. rubrum dec consequently rubrum that hypothesize we Therefore 2009). Holcomb, & Cohen (e.g. higher be to expected 2012; (Edmunds affected is metabolism the conditions pH lowered Under 1992). 1983, (Palmer conditions stress under construction shell the c the in described been has it As 2007). Tchernov, & (Fine conditions control in maintained those than higher times three was biomass their but skeleton, the of dissolution Articlecomplete a suffered species these of polyps case, this In 2007). the which in 7.4) (pH lowering corals pH temperate drastic more in found were responses Similar 2). (Fig. time with content matter organic in lowering progressive a displayed which control, the comparison in conditions pH lowered under reared colonies coral the in observed was matter organic of content higher experiment, the throughout example, For hypothesis. e Tsounis (Al consumption energy in increase consequent the with conditions, new the offset to processes physiological activated have may conditions mi to addition In observed. high of effect clear a intact, remained colonies of tissue the ditions in the present work were similar to those observed by Rossi & Tsounis (2007) in (2007) Tsounis & Rossi by observed those to similar were work present the in ditions Total carbohydrate, protein and lipid content in colonies grown under control control under grown colonies in content lipid and protein carbohydrate, Total may respond to decreased seawater pH be increasing the formation of organic matter, matter, organic of formation increasingthe be pH seawater decreased to respond may eaoim asd y n nrae n h cs o cliiain n lw pH low a in calcification of cost the in increase an by caused metabolism t al. 2012). Our results on Total Organic Matter tend to point in the direction of this of direction the in point to tend Matter Organic Total on results Our 2012). al. t

reasing calcification rates. This response is mediated by an alteration of alteration an by mediated is response This rates. calcification reasing . Patagonica O. vs

that needed to build the organic matrix is very different and changes and different very is matrix organic the build to needed that nimizing growth, the colonies of colonies the growth, nimizing dud e a. 02 ad h eegtc ot f acfcto is calcification of cost energetic the and 2012) al. et Edmunds

and arcs pharencis Madracis in situ in . Protein and lipid conten lipid and Protein . - Horani et al. 2003; Cohen & Holcomb 2009; 2009; Holcomb & Cohen 2003; al. et Horani ase of gastropods, the energy allocation for for allocation energy the gastropods, of ase

ee exposed were C. rubrum C. p CO 2

t was also comparable also was t reared under acidified under reared n oa got was growth coral in

Fn & Tchernov, & (Fine C. rubrum C.

to those in those to tissue in tissue C. C. C. © 2013 Blackwell Publishing Ltd Hoegh 2009; Peltzer & (Brewer reproduction or growth et Previati2002; (Rossi conditions adverse under survive to mechanism defensive a as observed Li 2011, al. et (Wong synthe protein in and & (Kurihara pH low under respiration in invested is energy the of part considerable reared a because probablyShirayama2004) specimens those in decreased also skeletogenesis mathei This urchins survive. sea on thus, findings and the with 2010) conforms al. et Previati respiration; (e.g. processes physiological certain under experimented the with disruption pressures, environmental cycle life of endogenous kind to some due or be dormancy could processes, This 2007). Tsounis and (Rossi conditions natural under pressur pH low under colonies the in trend seasonal dimmed more a of indication an be perhaps could response in difference This 3). (Fig conditions acidified the under colonies the in reduced more much was variability experimen the throughout observed was content lipid and protein carbohydrate, of concentration total the in variability larger a mentioned, As to counter the stress. effect ofexternal althoughazooxanth 2012b), al. et Gori 1991; of al. et Latyshev case 1974; Quinn the & (Meyers in anthozoans zooxanthellate described those to similar quite were esters acid fatty the of most but S4) time. first the of concentration fatty acid on Data 2006). al. et Rossi as such gorgonians other to

Acceptedas such processes for energy available less into translate could this Nonetheless, 2010). al. Article

hr te fertilizat the where Regarding composition, Regarding ellate organisms may have other physiological and metabolic mechanisms metabolic and physiological other may have organisms ellate sis related with basic metabolic paths as found in barnacle and copepods copepods andbarnacle in found as paths metabolic basic with related sis

Go 02. n oa seis mtbls rdcin a as been also has reduction metabolism species, coral In 2012). Gao & Paramuricea clavata Paramuricea o sces dvlpetl ae, avl ie ad sclerite and size, larval rates, developmental success, ion

aim of saving energy by reducing or shutting down some down shutting or reducing by energy saving of aim

ta toe n h cnrl ramn o te observed the or treatment control the in those than e a wide range of fatty acid profiles were observed (Fig. observed were profiles acid fatty of range wide a eietou pulcherrimus Hemicentrotus

and i te oto clne, hra the whereas colonies, control the in t C. rubrum C. Leptogorgia sarmentosa Leptogorgia - Guldberg et al. 2007). Our study was was study Our 2007). al. et Guldberg

are presented in this work forwork this arepresentedin

and

(Rossi 2002; (Rossi Echinodetra Echinodetra rc dfain o yas Te akt rc o te e crl oois s urnl vr hg: thin high: © 2013 Blackwell Publishing Ltd very currently is colonies coral red the of price market The years. for deflation price Acceptedindustry jewellery the of economy the on rebounds negative have could conditions, pH low against mechanism survival possible as colonies coral red of metabolism observed of response negative a time, first the for shows, study present The 2011). al. et (Joos century the of end the by significantly drop could states saturation grass2008, 2012;Tsou plain” (Rossiet al. “coral into forest” “coral the turning 2012b) al. et (Santangelo rules management by affected been have structures population deep and shallow Spain), (NE Creus de Cap In 2010). al. time over altered been have coralred of size and distribution the (SantangeloInfact, extinction et localal.2012a). populations to harvesting, some could bring uncontrolled the with coupled events, mortality mass These 2012). al. et Cupido 2007; al. e Santangelo 2009; 2001; al. et Garrabou 2006; al. et Coma 2000; al. et Cerrano 2005; al. et (Bramanti anomaly temperature a to due event mortality mass a suffered Sea, Mediterranean shallow Article 2003, and 1999 populations summer In change. environmental and climate rapid ongoing the underspecies this bestforstrategiesmanagement conservation implement and to essential and management 4.3 Economy coral red of response OA.populations to the on insights deeper gain to undertaken be should seasonal naturalenvironment a in response the on and reproduction on effects the on research further of rates growth skeletal on pH lower of effect the assessing on focused n h ohr ad dpnig n h CO the on depending hand, other the On Understandi

decrease in the calcification rate together with some evidence of a potential reduction in the in reduction potential a of evidence somewith together rate calcification the in decrease

of . rubrum C.

g h pplto dnmc o te rcos eierna rd oa is coral red Mediterranean precious the of dynamics population the ng aog te bnhc upnin edr i te northwestern the in feeders suspension benthic other among ,

nis etal.2007). 2

msin cnro, ewtr H n carbonate and pH seawater scenarios, emission C. rubrum C.

deep linked to this species and lead to a to lead and species this to linked (Rossi et al. 2008, Linareset 2008, al. et (Rossi - water populations water C. rubrum C.

. However, . to OA. The OA. to t © 2013 Blackwell Publishing Ltd Marie a by supported was L.B. pictures. the for Obis Eduardo to and sampling underwater for used equipment CCR Megalodon the for Ferrucci A. and TDI to due also are Thanks assistance. technical assist for Fortuño Manuel José thank We staff. Area Protected Marine Creus de Cap the of support invaluable the for grateful are authors The CTM2009 (MINECO; Competitiveness and Economy of Ministry Spanish researchThe funded been has by EC the 7 Aknowledgements precious species with associatedthe biodiversity andthe economy linked to its exploitation. of basis the on planned then be should actions conservation and Management scenarios. acidification future expected the under dynamics population on projections making allow will that 2001) (Caswell models matrix on based overtime study presentfromthe provide experimentalusedbe simulat inputin that can rubrum C. s populationthe in changes and extinction local Moreover, activities. anthropogenic other and overexploitation acidification, of populations local suffer may Sea Mediterranean of species precious represent more than 230 million US$yr Tsounis 2009; Muzik & (Nonaka Taiwan and is coralprocessedsignificantlythe(Italy)Greco indelonly overdeclined of Torreand, today,30% time i coral red Europe, In worldwide. decline a suffer may industry coral red the deflation, price possible and production material raw limited causing pressures, economic and environmental 50 reach can >4cm diameter 230 for sold be can branches juvenile

Accepted Articlerubrum C. Furthermore, without proper protection and management plans, this slow this plans, management and protection proper without Furthermore,

play in the benthic pelagic coupling and biogeochemical cycles (Rossi et al et (Rossi cycles biogeochemical and coupling pelagic benthic the in play , whereas 70 whereas , . rubrum C.

- 80% is represented by other species of precious corals imported fromJapanimported corals precious of species other byrepresented is 80%

tructure could affect the role that engineering species (Jones et al. 1994) as1994) al. etengineering (Jonesrolespeciesthat theaffect could tructure a b ual t cp wt cmie pesrs f et ae, ocean waves, heat of pressures combined with cope to unable be may .000 US$ kg US$ .000

- 1

(Tsounis et al. 2010). - ing us with SEM imaging and the ZAE staff at the ICM for ICM the at staff ZAE the and imaging SEM with us ing h otoe f hs smltos n re t peev this preserve to order in simulations these of outcome the 0 U$ kg US$ 300 th - 1

FP (grant agreementFP (grant (Tsounis et al. 2007, 2010). Under the increasing global increasing the Under 2010). 2007, al. et (Tsounis

t l 21; hn 02. hs eooi losses economic These 2012). Chen 2010; al. et - 1

hl Pcfc oalu s. oois with colonies sp. Corallium Pacific while

xicin n h na ftr. Young future. near the in extinction

265103 , Project MedSeA) andby the - ions of populationions of trends 84, rjc ACDC). Project 08849, - growing, endemic growing, . , 2012). Data 2012). , ndustry has ndustry

© 2013 Blackwell Publishing Ltd analysis variance. of Department Statistics, of University Auckland, of New Zealand. PERMANOVA:a (2005) MJ Anderson Ecology 26,32 non for method new A (2001) MJ Anderson doi:10.1007/9780114 Netherlands. Springer Dordrecht: Media. Eds.) Stambler, N. & Dubinsky (Z Reefs. to Cells Calcification, Coral (2011) S Tambutté and D, Zoccola É, Tambutté D, Allemand review. a Coral: Red Mediterranean the of skeletogenesis Precious Corals & Octocorals Research and biology The (1993) D Allemand 419 coral scleractinian the in respiration and photosynthesis Al Publishing andSpringer. Kingsford PA, Hutchings (eds management and environment biology, Reef: Barrier Great The In: Octocorals. (2008) KE Fabricius P, Alderslade 11: 36 InternationalBiophotonics ImageJ. with processingImage (2004) SJ Ram PJ,Magalhaes M, Abràmoff Refe GeneralitatdeCatalunya through grant2009SGR142. Global and Biogeochemistry Marine the from contribution 08 OCE (RyCContract Cajal y Ramón a byS.R.2009/No.3) UABEuropeanand no.JEMES scolarship (contract ErasmusMundus of by M.G. Curie

Accepted- Article Horani FA, Al FA, Horani – rences - 426.

4. - E flosi (EC7 fellowship IEF 42.

-

48 poie spot o .. uig pa during L.B. to support provided 44785

- 46. - Moghrabi SM, de Beer D (2003) The mechanism of calcification and its relation torelation its calcificationand mechanism of The (2003) D Beer de SM, Moghrabi

th

P Poet º 202, .. y a by J.M. 221072), nº Project FP, - 2007

- 01327) from MINECO. US National Scien National US MINECO.from 01327) 2 FORTRAN computer program for permutational multivariatepermutational for programcomputerFORTRAN ,19 – 39 - aaerc utvrae nlss f aine Austral variance. of analysis multivariate parametric J n Heh udeg ) p 222 pp. O) Guldberg Hoegh and MJ

t f h wiig f aucit Ti i a is This manuscript. of writing the of rt Galaxea fascicularis Galaxea

hne eerh ru, udd by funded group, research Change P suethp (BES studentship FPI aie B Marine . ce Foundation grantFoundation ce

- - 2007 4. CSIRO 245. iology - - 94 16537), - 007 142 - . , © 2013 Blackwell Publishing Ltd Caswell H (2001) Matrix Population Models, second ed. Sinauer Associates, Sunderland, MA. Méditerranée. la de gorgonaires Oceanographique Monaco, Les (1975) M Grasshoff C, Carpine Research Sea. Catalan the of case the ecosystems: marine Mediterranean on change climate of Effects(2011) C Pelejero JM, Gili SabatésA, J, M,Pascual Ribes ComaR, R, Simó E, Calvo acroporid corals. Journal of Experimental Marine Biology an D,Bucher Harriott RobertsVJ, LG (1998) Skeletal micro Mediterranean. NOAA Technical Memorandum CRCP International Workshop on Red Coral Science, Management, an Bussoletti E, Cottingham Bruckner D, AW, Roberts G, Sandulli R (2010) Proceedings of the trade, Hong Kong, 2009. NOAA Tech Memo NMFS Bruckner AW, Roberts G, (2009) 1st coral red BrewerLimits ET PG,Peltzer (2009) tomarine life. Science Mediterranean of case The monitoring: species ( emblematic in divers scuba recreational of Involvement (2011) G Santangelo S, Stolfa S, Rossi I, Vielmini L, Bramanti 2856. management gorgonians of corals: young andolds, could they coexist? Ecol Mod 20 (21): 2851 Bramanti L,Santangelo four a 1758), (L rubrum Corallium JournalMarine ofExperimental BiologyandEcology Coral Red Mediterranean the of growth the in Forcing Brama Anthropogenic and Sequestration CO2 , Ratios MediterraneanEnvironment ChemicalHdb Sea. Carbon and Oxygen Ruiz MS, Boukhary, JP, Bethoux, Detailed tissues: and animals marine in 12, 103 lipids Expe of of Journal lipids. total Estimationfor method sulphophosphovanillin the of (1973) investigation J Blackstock H, Barnes ocean of implications margin: the on 265 Life (2008) NR Mg Bates on acidification FT, Mackenzie AJ, Andersson Accepted rubrum Corallium Article – 273.

nti L, Magagnini G, DeMaio L, Santangelo G (2005) Recruitment, early survival and survival early Recruitment, (2005) G Santangelo L, DeMaio G, Magagnini L, nti – 118.

50

, 1 , - 29. -

). Journal for Nature Conservation calcite,

G, Iannelli M (2009) Mathematical modelling for conservation and

high latitude and cold and latitude high 71(

1), 1 Int Workshop on Corallium science, management and - Pino, D, Morin, P, Copin Montegut, C, (2005) Nutrient, (2005) C, Montegut, Copin P, Morin, D, Pino, - 140.

- water marine calcifiers. marine water - 5 OPR , 67 - 13, Silver Spring,13, Silver MD233 pp. - density, porosity andbulk densityof - - 19 43:71 , 86. 314 d Ecology , 312

d LessonsTrade: from the , 69 - 86 324 –

318. - 78. , 347. 228

, 117

Mar Ecol Prog Ser 373, Ser Prog Ecol Mar – ultn e Institut de Bulletin 136. rimental Biology rimental

- year study. study. year Climate Climate

-

© 2013 Blackwell Publishing Ltd marine ecosystem services. Kite S, Cooley 2165 10.1007/s00227 DOI Biol Mar acidification. ocean to rus Porites coral the of response (2013)Comeau EdmundsPJ Effects Carpenter S, offeeding and intensity light onthe RC, warming Global (2009) J Sciencesof Pascual J, Nati the of Salat Proceedings Mediterranean. the E, in events mortality Jiménezmass and stratification E, Serrano M, Ribes R, Coma Octocora (Cnidaria: singularis Eunicella of Mediterranean). Marine Ecology Progress Series 327, 51 mass populations a of Consequences (2006) in J Ballesteros J, mortality Garrabou D, Diaz M, Ribes C, Linares R, Coma doi:10.1371/journal.pone.0011842 Mediterranean Biodiversitythe (2010) al. The Raiset of Steenbeek PiroddiBenLasramF, C, Kaschner M, Coll K, J, Mineralogyin andGeochemistry, Co the Uncovering acidification: ocean about care corals Why mechanism. Oceanography, (2009) M Holcomb and AL Cohen 2129. m of calibration scale concentration measurements: pH seawater Spectrophotometric (1993) RH Byrne TD, Clayton Marine Policy 36(3) Chen CS (2012) Management of the precious coral fishery in Taiwan: Progress and perspectives meta ConnollyChan NCS, (2012) SR Sensitivity coral calcification toocean of acidification: 1999. summer Mediterranean), Letters (NW Sea Ligurian the in organisms other and gorgonians CN, Bianchi G, Bavestrello C, Cerrano

AcceptedReviewsmineralization. coral on perspectivegeochemical A (2003) McconnaugheyT and AL hen Article - -

analysis. Biology282 Global Change 19(1): 5 ,

3, 284 106 – 293. , 6176 - oel , Doney H, Powell e: siae, atrs ad het. LS N 58: e11842. 5(8): ONE PLoS Threats. and Patterns, Estimates, Sea:

:623 – 81. -

629. 22

(4), 118 Oceanography

54

(1), (1), 151 -

rsl upe n at and purple cresol (09. ca aiiiains oeta t atr global alter to potential acidification’s Ocean (2009). S – 127. t al. et

– 187. , 22

20) ctsrpi mass catastrophic A (2000) (4), 172 (4), - 290. – 60. - e rsls Deep results. sea - 181.

la i Mnra (NW Menorca in llia) - e Rsac 4, 2115 40, Research Sea - otlt eioe of episode mortality oa hdoe ion hydrogen total onal Academy onal - enhanced Ecology

a - 012

- - © 2013 Blackwell Publishing Ltd from recover Series and survive rubrum species Accepted coral i event mortality Mass (2001) JG Harmelin S, Sartoretto T, Scleractinian Perez J, Garrabou (2007) D decalcification. Tchernov M, Fine Porite Edmunds (2012) PJ Effect pCOof coralthe Edmunds Cumbo PJ, Fan VR, Progress In Oceanography,Mediterranean. 91(2), 97 the in al. forcings et anthropogenic C and climatic Estournel to F, responses D’Ortenzio ecosystems’ D, Marine Cossa (2011) P, Conan R, Sempéré C, Guieu X, Madron de Durrieu the for method Colorimetric (1956) F Smith PA, Rebers JK, determination of sugars and related substances. Hamilton KA, Gilles M, Dubois ocean for practices Annual Review Marine of Science best CO other the acidification: Ocean (2009) J to Kleypas R, Feely V, Fabry Guide S, Doney (2007) JR Christian CL, measurements. special PICES publication, 3. Sabine AG, Dickson Articleweighing techni Short (1989) PS Davies pelagicthe marine environment. St J, Dalsgaard output. Mar Ecol Prog Ser. press).(in reproductive quantifying population: gorgonian recovering reproductive, highly a of structure Sexual (2012) G Santangelo L, Bramanti M, Iannelli A, Peirano S, Ferrando V, Manno S, Cocito R, Cupido human multiple of effects cumulative and Interactive stressors marinein systems. Ecology 11: Letters 1304 (2008) BS Halpern K, Kroeker CM, Crain 269. characterization. genetic preliminary Sea: Mediterranean the from A, Remia M, Taviani F, Constantini

s spp. in Moorea, French Polynesia. Mar Biol 159: 2149 , 217 populations in the Provence region (France, NW Mediterranean). NW (France, region Provence the in populations Seriatopora caliendrum , 263

John M, Kattner G, Müller G, Kattner M, John - 272. que. Science

Marine Biology Marine ,

- 315 em rwh esrmns f oas sn a acrt buoyant accurate an using corals of measurements growth term

TY (2012)TY Metabolic costs of larval settlement and metamorphosis in , Advances in Marine Biology 1811.

under ambient and elevated pCO 2

on the growth, respiration, and photophysiology massive of , -

166. 1

, 169 ,

101 et al. et

- Navarra D, Hagen W (2003) Fatty acid trophic markers inmarkers trophic acid Fatty (2003) W Hagen D, Navarra - (3), 389 192. Analytical Chemistry (2010) Deep (2010)

- - 131 395. 5.

, - 46, - 2160 water water 225 2 . Biol Bull. Biol (in press).

, - 28 Corallium rubrum Corallium 340. , 350 Marine Ecology Marine

Marine Ecology Progress Ecology Marine – 356. n red coral red n

2

, (L.,1758) Corallium

problem. 31 , 261 , -

© 2013 Blackwell Publishing Ltd 5:213 Jeyasuria P,Lewis JC (1987) Mechanical properties of axial the skeleton gorgonians. in Coral Reefs and Ocean Acidification.Science 318,1737 Coral (2007) ME Hatziolos A, Dubi RH, Bradbury RH, Iglesias CM, Eakin N, Knowlton K, Caldeira AJ, Edwards PF, Sale CD, Hoegh to ecosystem perspective. organisman ecosystems: marinecalcifying in on organismsoceanacidification of effects The (2010) MA Sewel T, Klinger DA, Hutchins RD, Gates PJ, Edmunds JP, Barry GE, Hofmann (2008) V ecosystems. Fabry J, Guinotte Corallium rubrum Gol MC, Grillo (2012b) S Rossi L, Bramanti L, Magni C, shallow Cucio gorgonian versus deep M, in ecology Kotta trophic and JM, cycle Reproductive Gili N, Villadrich A, Gori DOI:10.1007/s00227 Mediterranean gorgonian (2012a) and of morphotypes thezooxanthellate azooxanthellate Characterization the of L,GoriLopez A,Bramanti 2010. SAC 13/2011/if. (2010) Report of the Transversal Workshop on Red Coral Alghero (Sardinia), Italy, 16 GF 5 France, (Corsica), 2011. Ajaccio Coral Red on Workshop Transversal the of Report (2011) SAC) Committee Advisory Scientific Mediterranean the for Commission Fisheries (General GFCM Global Change Biology 15,1090 o effects communities: benthic rocky Mass Mediterranean Northwestern (2009). in mortalityC Cerrano, F, Zuberer, M, Zabala, O, Torrents, N, Teixido, E, Serrano, JC, Romano, Harmelin, Gambi,JG, MC, D, Diaz, M, Cigliano, P, Chevaldonné, M, Bally, N, Bensoussan, R, Coma, J, Garrabou,

Accepted ArticleCM (General Fisheries Commission for the Mediterranean Scientific Advisory Committee SAC) –

219 - udeg , ub P, otn J Seek S Genil P Gmz , Harvell E, Gomez P, Greenfield RS, Steneck AJ, Hooten PJ, Mumby O, Guldberg

Eunicella singularis Annual New York Academy York Annual New dberg WM, Allemand D (1993) Skeleton and sclerite formation in the precious coral precious the in formation sclerite and Skeleton (1993) D Allemand WM, dberg Kersting, DK,Ledoux, Lejeusne,JB, C, Linares, C, Marschal, C, Ribes, Pérez, T, M, . Marine Biology -

012 - Eunicella singularis Annu. Rev. Evolu Ecology, 1928 - Gonzalez P, Thoma P, Gonzalez . Coral Reefs 31(3): 823 –

1103. - 3.

ca aiiiain n is oeta efcs n marine on effects potential its and acidification Ocean 117: 119

Sciences, - 128 - 1742.

. Mar. Biol. 159:1485 J, Gili JM,Grinyo UceiraJ, Gili S V,Rossi J,

1134, - 837 tion, andSystematicstion, .

320 Reefs Under Rapid Climate Change Climate Rapid Under Reefs –

ouain o te Mediterranean the of populations 342.

- 1496, - f the 2003 heat wave. heat 2003 the f Prieto R, Muthinga R, Prieto , 41, – 127 17 September – - 47. October 7

© 2013 Blackwell Publishing Ltd Octocorallia) . in sclerites surface of function The (1991) E Wallis von JC, Lewis Accepted Ecol Trends Sea. Mediterranean Evol 25(4):250 impacted highly diverse, highly the ocean: miniature a on effects Chevaldonn C, Lejeusne reef of acids Fatty (1991). YY Latypow VI, building corals. Svetashev NV, Naumenko NA, Latyshev atmosp increased of Effects (2004) development. Y Shirayama H, Kurihara Meta (2010). GG Singh, variable effects of ocean acidification on marine organisms. and Ecology letters, RN, Crim, RL, Kordas, KJ, Kroeker, 74, 4 Acta Cosmochimica et Geochimica acidification. and ocean Physiological to (2010) corals scleractinian A of Shemesh responses GL, isotopic Foster A, Meibom R, Yam M, Fine EJ, Hendy S, Krief a of Report Geological Survey, 88pp. research. future for guide 18 a held calcifiers: oceanworkshop marine of other Impacts and (2006) LL reefs Robbins coral CL, on Sabine acidification C, Article Langdon VJ, Fabry RA, Feely JA, Kleypas concentrations best predict calcification. Global Change Biology 16, calcification ofrates Jury CP, Whitehead RF, Szmant AM (2010) Effects of variations in carbonate chemistry onthe University Press, Oxford, pp. 272 ocean acidification projections, in:Ocean acidification Gattuso, (JP LHansson, eds.), Oxford F,FrJoos CJ,Jones Lawton ShachakJH, M(1994) Organisms ecosystem as engineers. Oikos 69:373 methodology Methods Research Franzisketgrowth:Coralbuoya Maragos(1978) PL, L JE, Jokiel 988 - 5001. ölicher SteinarcherTL, M, Plattner GK (2011) Impact climate of change mitigation on ,

Marine Ecology Progress Series pp 529

Biology Bulletin - 260. Mar. Ecol. Prog.Ser. Mar. Ecol. –

0 pi 20, t Ptrbr, L spo FL, Petersburg, St. 2005, April 20 es tdat R Jhne R) UEC mngah o oceanographic on monographs UNESCO RE), Johannes DR, Stoddart (eds Madracis auretenra -

542 é .

Paris. P, Pergent P,

- , 290. 181 - Martini C, Boudouresque CF, P CF, Boudouresque C, Martini

, 275

, (= 76 Madracis mirabilis - : 295 288. , 274, – 301. 161 –

69.

srd y S, OA ad h U.S. the and NOAA, NSF, by nsored

sensu Wells, 1973): bicarbonate nt weight technique.weightntIn: ei C2 n e uci early urchin sea on CO2 heric

1632 é - nlss eel ngtv yet negative reveals analysis rez T (2009) Climate change Climate (2009) T rez

- gorgonians (Coelenterate, gorgonians 1644. 13 (11), 1419

– Coral Reefs: Coral 34 – 386.

- © 2013 Blackwell Publishing Ltd Palmer AR (1992) Calcification in marine molluscs: how costly is it? Proceedings of Nationalthe Accepted from marine gastropods. Marine Biolog Palmer AR (1983) Relative cost producingof skeletal organic matrix versus calcification: evidence the twenty Totterdell, IJ, Weirig, MF, Yamanaka, Y,and Yool, A (2005) Antrop Najjar, RG, Plattner, GK, Rodgers, Sabine,KB, CL, Sarmiento, JL, Schlitzer, R, Slater, RD, Ishida, A,Joos,F, Key, RM, Lindsay, K, Maier C., Orr, Fabry,J VJ, Aumont, O, Bopp, L,Doney, SC, Feely, RA, Gnanadesikan, A, Gruber Ryukyu ArchipelagoMar Ecol Prog 397: Ser 269 Nonaka Muzik M, K (2009) Recent harvest records of commercially valuable precious corals in the corals, ProceedingsInternational ofthe 11thSymposium, Coral Reef Ft. scleractininan two of growth skeletal on temperature increased and acidification ocean of Effects (2008) PJ Edmunds N, Muehllehner accepted. non acidification. and ocean native to in response recovery and reduction Fernández E, Calcification Serrano M, Ribes R, Coma coral. C, Pelejero in E, Calvo J, acids Movilla fatty of analysis for method A (1974) N andOceanography,Limnology Marshall J, Quinn P, Meyers and global warming through pH acidification ocean to resilience ArticleCoral (2012) P Montagna J, Trotter J, Falter M, McCulloch reagent. Lowry elevated under truncata Myriapora bryozoan calcite Mg seawaterI: pCO2simulatingacidification. ocean PSZN Ecology Marine the in alterations geochemical Rodolfo C, Lombardi polymorphism i (2011a) S Cocito, P, Taylor C, Vasapollo MC, Gambi C, Lombardi SerProg 402: 69 areas and theconservation long of Linares C, Bianchimani O, Torrents O, Marschal C, Drap CO high a in more feeds and ocean. 1 bulletin pollution Marine respires producer secondary marine A (2012) K Gao W, Li

OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol Folin the with measurement Protein (1951) RJ Randall AL, Farr NJ, Rosebrough OH, Biological Chemistry

- first centuryfirst and its impact on calcifying organisms, Nature, 437, 681 - n bryozoans atnatural CO 79

- Metalpa R, Cocito S, Gambi MC, Taylor PD (2011b) Structural and and Structural (2011b) PD Taylor MC, Gambi S, Cocito R, Metalpa , 193

19 , 265 - up lived invertebrates:lived the Mediterranean red coral. Ecol Mar – , 846 ora o Eprmna Mrn Booy n Ecology and Biology Marine Experimental of Journal - 5. regulation.623 Change NatureClimate 2, y 292,287 –

275. - 848.

2

vents. - Reimer, R, E,Matear, Monfray, P,Mouchet, A,

- 278. – 292.

Zoomorphology

P, GarrabouP, J(2010) Marineprotected oilpr meandrina Pocillopora - aie eierna crl in corals Mediterranean native ogenic ocean acidification over Lauderdale, Florida. , 130 Skeletal alterations and alterations Skeletal , 135 32 – 686. : 211−222 - - Vallejo P (2012) P Vallejo 145. and – 627.

oie rus Porites

, N,

2 , .

© 2013 Blackwell Publishing Ltd Ecology: anevolutionary perspective 31:447 bryozoanthe temperatureson high andacidification AcceptedRodolfo 2100 AD. caespitosa Cladocora Rodolfo Open Survey Geological 2010 U.S. (iPhone): iOS and X, OS Max Windows, for calculator CO2calc (2010)SC Meylan JA, Kleypas HansenME, LL, Robbins CO to responsecalcification biological the interpreting for frameworkphysicochemical A (2011) JB Ries ocean acidification by the coral (2010) DC McCorkle and AL Cohen JB, Ries to CO exh calcifiers Marine (2009) DC McCorkle & AL, Cohen JB, Ries project.org Foundation for Statistical Computing, Vienna,Austria. ISBN 3 R Development Core Team (2007) R: a Article under different temperatures. ExperimentalJournal of Marine Biology andEcology 390,39 octocoralsMediterraneanin consumptionOxygen (2010) R, Osinga C, Cerrano A, Scinto M, Previati the dissolution of Mg to application data: rate from solubility mineral Predicting (1974) FT Mackenzie LN, Plummer carbon cycle models. F Joos R, Knutti GK, Plattner alkalinity determination. Perez FF, Rios AF, Rellán AlvarezT, M(2000) Marine Chemistry 21,169 Perez FF, Fraga F (1987) A precise and rapid analytical procedure for alkali Academy Sciences of the United States of America 89, 1379 2 - – induced oceaninduced acidification. 1280, 17p. 2 - - - induced ocean Geology,1131 acidification. 37(12), Metalpa R, LombardiR, Metalpa C, Ferrier S, Martin R, Metalpa

Biogeosciences,

-

calcites. American Jour Journal of Climate to long to Ciencias Marinas 26,463 - 182. 7 , 289 - term exposure to to exposure term et al. al. et

Oculina arbuscula

Geochimica et Cosmochimica Acta -

300. CocitoS,Hall (2008) Long (2008) -

Pages C, Gattuso JP (2010a) Response of the temperate coral temperate the of Response (2010a) JP Gattuso C, Pages language environment and for statistical computing. R ,

21 nal of Sciences, 274:61 , 2721 - 456. Improvements in a fast potentiometric seawater - p Spencer J, an Spencer J, nnier acfcto rsos t CO to response calcification nonlinear A

- – . Coral Reefs 29 (3): 661 - CO term climate commitments projected with climate with projected commitments climate term 478. 2751. Myriapora truncataMyriapora 2 n tmeaue ees rjce o h year the for projected levels temperature and

- d Gambid EffectsMC. (2010b)oceanof – 900051 – 82. 1134. – 83.

—

75 A user A

-

, 4053 07

at natural COnaturalat ibit mixed responses mixed ibit - - nity determination. 0. 674. - friendly seawater carbonseawater friendly – http://www.R 4064.

2

vents. Marinevents. - ie Report File – - 48. 2 - induced

- © 2013 Blackwell Publishing Ltd of Patterns Corallium rubrum (2012b) JM Gili C, Lott I, Vielmini recruitment variation in post and G, Tsounis S, Rossi L, Bramanti G, Santangelo Coral ReefCairns, Australia, Symposium, 9 long of Demography (2012a) M Iannelli G, Tsounis L, Bramanti S, Cocito R, Cupido G, Santangelo of theover biology conservation and dynamics Population (2007) M Iannelli L, Bramanti G, Santangelo 204. acid fattyMediterran in over the of Demography (2004) columns L Bongiorni L, Bramanti E, Maggi extraction G, Santangelo phase solid of Use The (2007) purification. J Werne J, Russell method for analysis of lipid fractions musclein foods. AntequeraRuiz J, AndresT, AI, MJ, Petron Muriel E (2004) Improvement a of solid phase extraction 97 population dynamics in a S,BramantiRossi L, Broglio E,Gili JM (2012). Trophic impact of long Mediterranean). deep of Survey Rossi in S, Rossi in protein, evidencesfor feeding summer autumn constraints. of concentrations cycles Seasonal lipid (2006) N Vert and A, Gori carbohydrate C, Linares R, Coma JM, Gili S, Rossi feeders. suspension benthic PhD Thesis, University of Barcelona. of ecology trophic the affecting factors Environmental (2002) S Rossi acidification adversely affected by warming. Climate Nature Change, 1,308 – Accepted Article rubrum Corallium 111, doi: 10.3354/meps09848

, sui G Oea C Pdó T Gl J, rmni , exd N Gt J ( J Gutt N, Teixidó L, Bramanti JM, Gili T, Padrón C, Orejas G, Tsounis S, - - lived octocorals: survival and local extinction. Proceedings of the 12th International 12th the of Proceedings extinction. local and survival octocorals: lived Tsounis G (2007) Temporal and spatial variation in protein, carbohydrate, and lipids lipids and carbohydrate, protein, in variation spatial and Temporal (2007) G Tsounis - ean red coral ( coral red ean exploited Mediterraneanexploited coral. red Organic Chemistry - wlig e crl ( coral red dwelling . J Exp Mar Marine Biology Marine (Anthozoa, Octocorallia)

short Corallium rubrum Corallium Biol Ecol 411: 7 - lived hydrozoan, - recruitment processes of the Mediterraneanrecruitmenttheprocesses gorgonianof precious coral

, , 38 154

(1), 48 oalu rubrum Corallium , 533 - – 51. - - 545. 13. aauie clavata Paramuricea 13 July 201212A 13 July Eudendrium racemosum L. 1758). 1758). L.

Marine Biology, Marine

Journal ofTheoretical BiologyJournal

. (2011) Coral and Coral (2011) . Anal Chem Acta520:201 Marine Biology ) populations at Cap de Creus (NW (NW Creus de Cap at populations ) Scientia Marina, Scientia Life

157 - lived species indicated by (nhza octocorallia): (anthozoa, : , - , 429 149 Mar Ecol Prog Ser. 467: ocean to resistance mollusk history and reproduction. historyand – 312. , 643 - - 439. 205. 68

–

(Suppl. 1), 199 1), (Suppl. 651. ,

24 4,

- 416 exploited 2008) – 423. -

© 2013 Blackwell Publishing Ltd Review In: corals. precious of exploitation The (2010) conservation JM Gili and L, Bramanti (NW G, Santangelo R, Brava GriggAccepted S, Rossi Costa G, Tsounis the at fishery coral Red (2007) W Arntz JM, Gili Mediterranean): case study an for over harvested prec S, Rossi G, Tsounis ScienceLetters, 303,163 caespitosaCladocora etTrotter al.(2011) J Quantifying'vitaleffect' thepH inthe temperate zooxanthellate coral Marine BiologyEcology, and357,7 assessing Mediterranean.NW effects ofExperimental thepotential in Journal ofwarming vs. deep ofthe populations precious red Mediterranean Torrents N,Garrabou E, Caminiti (2008) O,Tambutte Upperofshallow J thermal thresholds the on Transient Sea.I: DeepPart Research Sea Oceanographic Papers Research Mediterranean Eastern the of CO anthropogenic Impact of distribution (2011) C Goyet and F, Touratier 1708 Mediterraneanthe mid from Sea TouratierGoyet andDecadal (2009) of evolution anthropogenicCO Articleworld.Biology GlobalChange 18,3015 La and ME Warner VR, Johnson Hall DJ, Suggett and temporalEarth patterns. AM,Key GordonSmith mineralogy DP(2006) MMJr, Skeletal bryozoans: Taxonomic of 190, Ecology and Biology Marine Experimental of Journal compounds. antifouling for evidence corals: soft Antarctic in defenses Chemical (1995) JN Heine and JB McClintock M, Slattery thein eastern Me contentcarbon anthropogenicHigh (2010) DWR Wallace and A, Körtzinger T, Tanhua A, Schneider Research Letters Geophysical Sea. Mediterranean the of Alkalinity (2007) A Körtzinger DWR, Wallace A, Schneider 61 – 1716 - (eds 77.

Gibson R, Atkinson R, and R, Atkinson GordonGibson R, J diterranean, Geophys. J. Res., 115,C12050, doi:10.1029/2010JC006171. 34 - pne J, Rodolfo JM, Spencer , 1 - 5. : Validation of theboron Planetary seawaterand Earth pHproxy.

- 173. - Sci. Rev.78:287

- 1990s to themid 1990s to 2 sn (02 Sa nmns a trv i a ih CO high a in thrive may anemones Sea (2012) T wson

n frt siae o estimate first and – 19. - -

3025, doi: D, Pettay Tye R, Payton TG, Boatman R, Metalpa caorpy n Mrn Booy A Annual An Biology: Marine and Oceanography - 306. ious coral.

- ), 2000s, Deep Sea Research Part I,2000s, Deep Part Research Sea 56,

10.1111/j.1365 48, coral 161 aiiiain o te Mediterranean the for acidification f Ecosystems - 212. Corallium rubrum Corallium 58

, 1 - 2486.2012.02767.x 2

- in the northwestern , 15. 10,

975 – 986.

(L.):

2

© 2013 Blackwell Publishing Ltd Accepted Fig S1.Colony of Figure captions (Integrated acidifica Marine Biogeochemistry andEcosystem Research), Newsletter, Issue n.#20,May 2012. to turns Research (2012) P Ziveri Sea 124. Black pp. & 2009, 36, Mediterranean Vol. Series, the Monograph (CIESM), Committee in Science Mediterranean Systems Mediterranean, Physical and Chemical Biological, on Acidification Jeffree R,Montagna P,Rees AP, ReynaudRodolfo S, C, Goyet M, Gehlen F, Gazeau JP, Gattuso SW, Fowler M, Fine S, Dupont G, Lange De A, Yilmaz atbut a cost. Proc. R.Soc. B 275,1767 calcif increase may acidification Ocean (2008) S Widdicombe and JI Spicer HL, Wood Proteomics 6: 311 CO2 htt Thiyagar PTY, Leung AC, Lane KKW, Wong Article Mar.Policy. high a in invertebrates Marine Biology AnnualReview, Benthic (2012). JM Roberts and L Wicks rubrum. architecture of red coral ( biomineral macroscale to Nano (2008) C Marschal O, Grauby A, Baronnet J, Garrabou D, Vielzeuf Corallium the from northwesternspecies Mediterranean ascidian several of survival and Growth (1992) MA Becerro of and X Turon p://dx.doi.org/10.1016/j.marpol.2012. sustainable harvesting for hurdles Management (2013) G Santangelo L, Bramanti S, Rossi G, Tsounis (2012). S. Rossi A, Martinez JM, Gili N, octocoral Viladrich Mediterranean the L, in reserves energy of allocation and Bramanti effort reproductive on effects Anthropogenic L, Martinez G, Tsounis

- driven seawater acidification. Comparative Biochemistry and Physiology Part D: Genomics and Genomics D: Part Physiology and BiochemistryComparative acidification. seawater driven Paramuricea clavata

- Corallium rubrum 321.

Corallium rubrum . Mar Ecol. Mar Prog Ser 82: 235 . MEPS 449:161 50

, 127

in the experimental aquarium – 1773 doi:10.1098/rspb.2008.0343 – 12.010i 188. ). American). Mineralogist 93: 1799 in n wrig n h Mdtraen e, IMBER Sea, Mediterranean the in warming and tion

ajan V (2011) Response of larval barnacle proteome toproteome barnacle larval of Response (2011) V ajan - 172

- Metalpa R,Ziveri P,Bri - 247.

- CO 2

ol. caorpy and Oceanography world. - and Impact F(2008) of 1815

ication rates, ication

© 2013 Blackwell Publishing Ltd un (n=24). colonies mg (µg concentration acid fatty Total 4. Fig. respectively (mean SD). ± coefficientCV= of variation. mg (µg sampled. were concentration (c) carbohydrate and (b) of coenenchyma lipid (a), protein Total 3. Fig. units), respectively (mean ± SD). experiment. whole coenenchymaof the in % matterorganic Total 2. Fig. gram weight initial of per day. n=15,error bars represent SE. pH bars, (black conditions control of rates Calcification 1. Fig. Acids; PUFA: Poly Unsaturated fatty acids. Fig. S4.Fatty acids composition. observed in the of images (SEM) Microscope Electron Scanning of Examples S3. Fig. respectively (three replicates per treatment); J) Microbubble diffusers. CO kg 50 G) filter; lime soda F) valves; E logger; data and controller pH D) measurements; temperature for probes PT100 and pH pH conditioningat seawater for tanks l 150 large setup Experimental S2. Fig.

Accepted its), respectively (mean ± SD). CV= coefficient variation. of Article C. rubrum. Black and grey bars represent control (pH control represent bars grey and Black Corallium rubrum Corallium

lc ad ry as ersn cnrl (pH control represent bars grey and Black Black and grey bars represent control (8.10 pH (8.10 control represent bars grey and Black

C. rubrum C. used to control and modify seawater pH in each aquarium. A) and B) and A) aquarium. each in pH seawater modify and control to used

SAFA: total Saturated Fatty Acids; MUFA: Mono Unsaturated Fatty

T

8.10 units). Calcification rates are expressed as mg CaCO mg as expressed are rates Calcification units). 8.10 colonies (n = 24) during all the periods of time when colonies when time of periods the all during 24) = (n colonies

after 314 days under lo under days 314 after 2

bottle; H) and I) control and low pH experimental aquaria, experimental pH low and control I) and H) bottle; - 1 T

7.81 and 8.10, respectively; C) glass electrodes forelectrodes glass C) respectively; 8.10, and 7.81 OM) in the coenenchyma of of coenenchyma the in OM) Corallium rubrum Corallium

T

8.10 units) and treatment (7.81 pH (7.81 treatment and units) 8.10

T w pH w

.0 nt) n tetet 78 pH (7.81 treatment and units) 8.10 T T

units) and treatment (7.81 pH (7.81 treatment and units) (grey bars, pH bars, (grey

colonies (n = 24) during the during 24) = (n colonies he tps f sclerites of types three oalu rubrum Corallium T

- 1 7.81 units) and units) 7.81

M i the in OM) ) solenoid ) T

units), 3

per T T

© 2013 Blackwell Publishing Ltd Accepted Article

© 2013 Blackwell Publishing Ltd Accepted Article

© 2013 Blackwell Publishing Ltd Accepted Article