Hvordan påvirker CO 2 post-smolt i brakkvann RAS?

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Εἰλείθυια Γιαννόπουλος
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1 Hvordan påvirker CO 2 post-smolt i brakkvann RAS? Tom O. Nilsen,. Mota, V.C., Kolarevic, J., Ytterborg, E., Ebbesson, Lars O.E., Handeland, S.O., Stiller, K., Terjesen, B.F SUNNDALSØRA, 12. NOVEMBER 2018

2 Bendik Fhyn Terjesen Vasco Mota Thanks to Michele Gallo, Jascha Gerwins, and all user partners, researchers who have contributed.

Problem definition Dissolved CO 2 accumulates in RAS Fish

oxygen consumption Higher: Nephrocalcinosis (deposits in

acidosis Altered gene expression Safe levels for Atlantic

3 Problem definition Dissolved CO 2 accumulates in RAS Fish exposed to high CO 2 shows: Lower: Feed intake, growth and oxygen consumption Higher: Nephrocalcinosis (deposits in kidneys) & cataracts Impaired osmoregulation and blood acidosis Altered gene expression Safe levels for Atlantic salmon: < mg/l Is tolerance for CO 2 in brackish RAS different? 3

4 Experimental design Reciprocal CO 2 exposure Water quality: - Oxygen >85 % - Salinity 12 ppt - Temperature C - ph Alkalinity mg/l -3 wk 0 wk 3 wk 6 wk 12 wk 6 wk Acclimation Chronic CO 2 exposure Seawater phase (common garden) Stocking 50 fish/tank Sampling 5 fish/tank Sampling 5 fish/tank Sampling all fish Sampling 5 fish/tank Sampling 5 fish/tank 4

5 5

6 Growth (SGR) Negative effects of elevated pco 2 on fish growth 10% growth reduction for every 10 mg/l increase in pco 2 Negative growth effects of elevated pco 2 in RAS persisted following transfer to seawater 5mg/L 19mg/L -12% 40mg/L CO2 concentration

7 Health & welfare No observed effects on: hematocrit glucose eye cataracts welfare score (skin & fins) nephrocalcinosis Photos: NOFIMA

8 Skin thickness (μm) Health & welfare 12 weeks in RAS 5 mg/l 100 μm 40 mg/l Dermis Epidermis Scale 100 μm 8 Mota et al., 2019

9 Physiological responses 12 weeks in RAS pco 2 plasma vs. water Choride vs. bicarbonate 9

10 Physiological responses - long term Plasma bicarbonate Plasma chloride 5 mg/l 12 mg/l 19 mg/l 26 mg/l 33 mg/l 40 mg/l 10

RAS 40 FTS 40 RAS 5 FTS 5 40R1>3100.diff_corr 40R2>3101.diff_corr 40R3>3102.diff_corr 40R4>3103.diff_corr 40R5>3104.diff_corr 40R6>3105.diff_corr 40S1>3106.diff_corr 40S2>3107.diff_corr 40S3>3108.

diff_corr 5S2>3119.diff_corr 5S3>3120.diff_corr 5S4>3121.diff_corr 5S5>3122.diff_corr 5S6>3123.

11 RAS 40 FTS 40 RAS 5 FTS 5 40R1>3100.diff_corr 40R2>3101.diff_corr 40R3>3102.diff_corr 40R4>3103.diff_corr 40R5>3104.diff_corr 40R6>3105.diff_corr 40S1>3106.diff_corr 40S2>3107.diff_corr 40S3>3108.diff_corr 40S4>3109.diff_corr 40S5>3110.diff_corr 40S6>3111.diff_corr 5R1>3112.diff_corr 5R2>3113.diff_corr 5R3>3114.diff_corr 5R4>3115.diff_corr 5R5>3116.diff_corr 5R6>3117.diff_corr 5S1>3118.diff_corr 5S2>3119.diff_corr 5S3>3120.diff_corr 5S4>3121.diff_corr 5S5>3122.diff_corr 5S6>3123.diff_corr Gene expression, microarray Gill: 40 mg/l and 5 mg/l treatments Low DEG / gene suppression in RAS Genes Tpr protein - Ident Ankycorbin Dystonin Tektin Keratin_ type I cytoskeletal 18 - Ident Tubulin--tyrosine ligase-like protein Exosome complex exonuclease RRP4 [Salmo salar] Dexamethasone-induced Ras-related protein precursor [Salmo salar] Rho GTPase-activating protein AP-1 complex subunit gamma-like 2 [Salmo salar] Diaphanous Telomerase-binding protein EST1A E3 ubiquitin-protein ligase MARCH MHC class I antigen [Salmo salar] TAP2b [Salmo salar] CC chemokine with stalk CK2 [Oncorhynchus mykiss] C-C motif chemokine 8 precursor [Salmo 0.30 salar] C-X-C chemokine C-X-C chemokine Complement component C7 [Salmo salar] Complement component C8 gamma chain 1.57precursor [Salmo 0.97 salar] SAPS domain family member 3 [Salmo-1.29 salar] Small inducible cytokine A13 [Oncorhynchus mykiss] Small inducible cytokine A13 [Oncorhynchus mykiss] putative interferon-alpha/beta receptor 0.12 alpha chain [Oncorhynchus mykiss] Complement component 9, Perforin precursor fish virus induced TRIM protein [Oncorhynchus mykiss] Gig Gig Gig ISG15-like Ubiquitin protein ligase E3A, hect domain ZNF C-type lectin domain family 4 member 0.56 E [Salmo -0.89salar] C-type lectin M4, cd209l Fish-egg lectin [Salmo salar] Rhamnose binding lectin STL2 [Oncorhynchus mykiss] Rhamnose-binding lectin precursor [Salmo salar] Growth factor independent Matrix metalloproteinase modified T cell receptor alpha [Salmo salar] TNF receptor member 11B S100 calcium binding protein A10b aminolevulinate synthase, erythroid-specific, mitochondrial precursor [Salmo salar] Cytidine deaminase [Salmo salar] Ectonucleoside triphosphate diphosphohydrolase [Salmo-0.57 salar] Carboxypeptidase A1 precursor [Salmo 0.43 salar] Nucleolar protein 5 [Salmo salar] Poly polymerase 10 [Salmo salar] Fatty aldehyde dehydrogenase [Salmo-1.52 salar] Sulfotransferase family cytosolic 2B member [Salmo salar] Claudin-like protein ZF4A22 [Salmo salar] Fibulin-1 [Salmo salar] Nephronectin variant 2 - Ident Otoancorin Glucocorticoid receptor Inactive serine protease PAMR Solute carrier family 13 member 3, slsc13a Tripartite motif-containing protein [Salmo-1.56 salar] homeobox protein HoxC8ba [Salmo salar] Lim homeobox protein 3 [Salmo salar] Extracellular matrix protein 1 precursor [Salmo salar] leukolectin protein [Salmo salar] leukolectin protein [Salmo salar] Up-regulator of cell proliferation Col6a2 protein - Ident GMP Giant mucus protein Calcitonin-1 precursor [Salmo salar] growth hormone receptor isoform 1 precursor [Salmo salar] Angiogenin-1 precursor / RNase ZF G-protein coupled receptor UDP-GlcNAc:betaGal beta-1_3-n-acetylglucosaminyltransferase-like Ident Optineurin [Salmo salar] Synaptobrevin homolog ykt6 [Oncorhynchus mykiss] Synaptosomal-associated protein 25-A [Salmo salar] Zymogen granule membrane protein precursor 0.06 [Salmo salar] Zymogen granule membrane protein precursor [Salmo salar] Ceacam EGF-like domain-containing protein 7 precursor [Salmo -2.49salar] Fibronectin 1b - Ident Leucine rich repeat containing 8 family_ member C Tetraspanin-1 [Salmo salar] Uncharacterized protein Uncharacterized protein Uncharacterized protein Unknown Ankyrin repeat and SAM domain-containing protein Immunoglobulin superfamily member Immune genes Other 11

12 Experimental design Reciprocal CO 2 exposure Water quality Oxygen >85 % Salinity 12 ppt Temperature C ph Alkalinity mg/l -3 wk 0 wk 3 wk 6 wk 12 wk 6 wk Acclimation Chronic CO 2 exposure Seawater phase (common garden) Stocking 50 fish/tank Sampling 5 fish/tank Sampling 5 fish/tank Sampling all fish Sampling 5 fish/tank Sampling 5 fish/tank 12

13 Post-smolts rapidly acclimate to large changes in pco 2 Rapid changes in pco 2 elicits clear transient stress responses measured as plasma cortisol and glucose. 5 mg/l 40 mg/l 5 to 40 mg/l 40 to 5 mg/l Plasma cortisol Plasma glucose 13

14 Post-smolts rapidly acclimate to large changes in pco 2 Rapid changes in pco 2 elicits clear transient stress responses measured as plasma cortisol and glucose. 5 mg/l 40 mg/l 5 to 40 mg/l 40 to 5 mg/l Plasma cortisol Plasma glucose 14

15 Post-smolts rapidly acclimate to large changes in pco 2 Rapid changes in pco 2 elicits clear transient compensatory responses measured as plasma HCO3 - and chloride. 5 mg/l 40 mg/l 5 to 40 mg/l 40 to 5 mg/l Plasma HCO3 - Plasma chloride 15

16 Summary and conclusions No mortalities, cataracts, nephrocalcinosis or poor external welfare observed in fish exposed to CO 2 from 5 40 mg/l Classic compensatory response (HCO3 & Cl) to elevated pco 2 Atlantic salmon very resilient to high CO 2 Elevated CO 2 has a growth penalty, and this penalty starts at lower concentrations than previously reported Ongoing work is focusing on physiological mechanisms and interaction of acid-base and ion regulation in smolts and postsmolts coupled to water qualities 16

17 Takk for oppmerksomheten TOM OLE NILSEN

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