Effects of water salinity and exercise on Atlantic salmon performance as postsmolts in land-based closedcontainment

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Ytrestøyl 1, J. Kolarevic 1, S. Calabrese 2,3, B.O. Rosseland 4, H-C.

1 Effects of water salinity and exercise on Atlantic salmon performance as postsmolts in land-based closedcontainment systems B.F. Terjesen 1*, T. Ytrestøyl 1, J. Kolarevic 1, S. Calabrese 2,3, B.O. Rosseland 4, H-C. Teien 4, Å. Åtland 5, T.O. Nilsen 2, S. Stefansson 2, S.O. Handeland 6, Jorn Schoordik 7, jh. Takle 1 1 Nofima, 2 Univ. of Bergen, 3 Marine Harvest Norway, 4 Norw. Univ. of Life Sciences; 5 NIVA; 6 UNI Research; 7 Hoogeschool Zeeland AIW#5 1

Background Loss of fish in Norwegian production at same level now, ~20% during cage phase, as 12 years ago (Arealutvalget 04/02/11) Much of the loss of fish, and

, 2010) Mass specific feeding rate of 1+ salmon (SFR, % day -1 ) stocked at sea late May Oehme, M., Grammes, F., Takle, H., Zambonino-Infante, J.-L., Refstie, S.

2 Background Loss of fish in Norwegian production at same level now, ~20% during cage phase, as 12 years ago (Arealutvalget 04/02/11) Much of the loss of fish, and feed intake and growth reduction, occur early after stocking small smolts in sea (<100 g/ind) (e.g. Oehme et al., 2010) Mass specific feeding rate of 1+ salmon (SFR, % day -1 ) stocked at sea late May Oehme, M., Grammes, F., Takle, H., Zambonino-Infante, J.-L., Refstie, S., Thomassen, M.S., Rørvik, K.-A., Terjesen, B.F., Dietary supplementation of glutamate and arginine to Atlantic salmon (Salmo salar L.) increases growth during the first autumn in sea. Aquaculture. 310, /11/2013 2

Background cont d Lice problems are considerable in cage production One possible solution can be to reduce the time spent in traditional open cages in sea The Ministry of Fisheries and Coastal

3 Background cont d Lice problems are considerable in cage production One possible solution can be to reduce the time spent in traditional open cages in sea The Ministry of Fisheries and Coastal Affairs in Norway recently opened for an increase in fish size in land-based facilities, from 250 g to 1 kg A larger smolt for stocking at sea, may reduce lice-problems, may reduce loss of fish, and improve growth may reduce prod. time, especially in areas with low winter temp Several Norwegian companies and R&D institutions joined forces in 2012 to explore how such post-smolt production can be done, in the RCN project OPP Optimized Postsmolt Production Aerial photo: Erik Røed 9/11/2013 3

4 Main objective in OPP To develop basic and applied knowledge about how the time spent in open cages in sea can be reduced, by increasing the time on land or in semi-closed systems at sea, and to determine the consequences for the performance, physiology, and welfare of the fish and production costs 9/11/2013 4

before slaughter 0-70 g 70 g 1 kg 1 kg 5 kg Semi-closed containment in sea Smolt prod in freshwater RAS

5 Combination-lines, to 1 kg in closed-containment systems Postsmolt in seawater? RAS landbased Closedcontainment on land Smolt prod in freshwater RAS landbased Shorter period in cages before slaughter 0-70 g 70 g 1 kg 1 kg 5 kg Semi-closed containment in sea Smolt prod in freshwater RAS landbased Postsmolts in semi-closed containment in sea Shorter period in cages before slaughter 0-70 g 70 g 1 kg 1 kg 5 kg

OPP: Problems to tackle Problem: Cato Lyngøy, former Group Technical Manager Technology & Environment Marine Harvest, and chairman in OPP: «So far we have not seen any closed-containment system that

requirements of Atlantic salmon postsmolts are in closed containment systems, for optimal technology performance, fish performance, welfare, and health To be tested in OPP: Land-based RAS

6 OPP: Problems to tackle Problem: Cato Lyngøy, former Group Technical Manager Technology & Environment Marine Harvest, and chairman in OPP: «So far we have not seen any closed-containment system that is sufficiently cost-effective, for production of a postsmolt up to 1 kg, neither on land or in sea» (IntraFish, 29/10/12) Problem: Insufficient biological knowledge of what the physiological requirements of Atlantic salmon postsmolts are in closed containment systems, for optimal technology performance, fish performance, welfare, and health To be tested in OPP: Land-based RAS (recirculating aquaculture systems) for production to 1 kg Nofima Centre for Recirculation in Aquaculture, Sunndalsøra, Norway To be tested in OPP: Production in semi-closed containment systems in sea to 1 kg ( m 3 vol, 450 m 3 /min flow) Photo: Aquafarm Equipment 9/11/2013 6

7 Salinity in land-based RAS, combination-line SW-RAS may have higher running costs than FW-RAS due to: CO 2 removal efficiencies are lower in SW than in FW (e.g. Moran, 2010) Ammonia removal is lower in SW compared to FW (e.g. Chen et al, 2006) Results in need for larger installation and/or higher flow Or can postsmolts be kept at lower salinity in RAS, and still handle full-strength SW at stocking in sea? Nofima Centre for Recirculation in Aquaculture (NCRA), Sunndalsøra, Norway 9/11/2013 7

8 Objectives and experimental design, effects of salinity and exercise for postsmolt in RAS What is optimal salinity for postsmolts in RAS, in terms of survival, health, and maturation to 1 kg size? Can lower salinity reduce maintenance costs and increase available energy for growth? Can exercise through water velocity contribute to these factors, and interact with salinity treatment? 2 + Exercise 12 -RAS 250g 12 -RAS 450g 2 - Exercise 12 -RAS 250g 12 -RAS 450g Parr ifw Nofima NCRA 12: L 2 + Exercise FT -Exercise 32 -RAS 250g 32 -RAS 32 -RAS 250g 32 -RAS 450g 450g 2 + Exercise 22 RAS 250g 22 RAS 450g 2 - Exercise 22 RAS 250g 22 RAS 450g 70g 250g 450g 800g 9/11/2013 8

9 Methods 3 separate RAS with salinities 12, 22 and 32 ppt, and two exercise levels, i.e. a 3x2 factorial study 7 200, 70 g smolts were stocked in 12 tanks (3.2 m 2 ), duplicate/treatment 12:12h light:dark during the experiment, C, high marine protein commercial diet (Havsbrun, 3-4 mm pellet), feed intake measurements On-line measurements of e.g flow, ph, ORP, temperature, O 2 A range of tissue samples collected, for trad. physiology, histology, and molecular physiology

RAS configuration during trial RAS 2 5 Ozone side-stream Pump Pump sump w/overflow 6 9 Belt filter w/sludge discharge 4 Ozone injection w/degas separator Make-up 11 water with flowmeter Pump sump

10 RAS configuration during trial RAS 2 5 Ozone side-stream Pump Pump sump w/overflow 6 9 Belt filter w/sludge discharge 4 Ozone injection w/degas separator Make-up 11 water with flowmeter Pump sump w/overflow 11 8 Degasser 9 10 Moving bed bioreactor 7 Central water treatment room 12 Oxygenation 2 1 Swirl separator w/sludge discharge Fish tank 3 Sidewall drain Fish tank Fish tank Experimental hall 1/2/

Swimming speed (BL/s) to ~250 g Body lengths/sec 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0 01.okt.

11 Swimming speed (BL/s) to ~250 g Body lengths/sec okt.12 Mean ± range (n=2 tanks per treatment) 15.okt.12 32ppt minus exercise 12ppt minus exercise 22ppt minus exercise 32ppt plus exercise 12ppt plus exercise 22ppt plus exercise Flow 85 l/min Flow 85 l/min 29.okt nov.12 Flow 132 l/min Flow 132 l/min 26.nov.12 Date Exp. day 10.des des jan Direction of the water flow Tank inlet % of total tank width Top view 9% 35% 63% % of total tank depth Lateral tank view

12 RAS conditions RAS Temp C ph CO 2 mg/l Alkalinity CaCO 3 mg/l % Reused flow % Water exchange /day Feedload, % of capacity Feedload/ water exh (kg/m 3 /day) ± ± ±1.5 72± ±0.8 25±7 11.0± ± ± ± ± ± ±0.7 24±6 10.6± ± ± ± ± ± ±0.4 25±2 9.1± ±

13 Water quality: TAN and NO 2 -N concentration TAN concentration (mg/l) Average TAN conc. at the tank outlet NO 2 -N conc. (mg/l) Average NO 2 -N conc. at the tank outlet

TAN removal efficiency over MBBR TAN removal efficiency (%) 60 50 40 30 20 10 0 36 34 0 38 49 22 21 13 04.12.12 03.01.13 07.05.

14 TAN removal efficiency over MBBR TAN removal efficiency (%) MBBR TAN removal efficiency (%) MBBR inlet TAN water conc. (mg/l)

CO 2 removal efficiency over countercurrent, forced-ventilated degasser Total CO 2 removal efficiency (%) 100 80 60 40 20 0 57 63 34 63 73 75 70 28 04.12.12 03.01.13 26.02.13 07.05.

15 CO 2 removal efficiency over countercurrent, forced-ventilated degasser Total CO 2 removal efficiency (%) CO2 inlet concentration (mg/l) CO2 removal efficiency vs CO2 inlet concentrations CO2 removal efficiency (%)

16 Postmolt sampled at ~250 g, produced in 12 ppt S RAS TGC to~250 g/ind Ind. weight at ~250 g/ind training 2.6 no training Thermal growth coefficient 250 g bodyweight (g) Salinity: p 0.01 Training: p ppt 22 ppt Training No training Factor salinity: p< Faktor exercise p< Salinity*exercise: p< ppt Salinity test 16 32

17 Postmolts sampled at ~800 g, produced in RAS Final weight TGC entire experiment Salinity: p 0.01 Training: p 0.01 training no training 12 ppt 22 ppt 32 ppt Thermal growth coeffcient Exercise No exercise Factor salinity, p<0.01 Factor exercise, p<0.01 Salinity, ppt

18 Cardiosomatic index (% BW-1) Results: Organ indexes at 250 and 800 g Heart index 800 g/ind Heart index 250 g/ind Exercise No exercise Faktor salinitet: p< Faktor trening: p< Male gonad size 250 g/ind 0.12 Exersise No excersise 0.10 Faktor salinitet: NS Faktor trening: NS Salinity 32 Gonadosomatic index (% BW-1) Gonadosomatic index (% BW-1) Salinity Male gonad size 800 g/ind 0.12 Exersise No excersise Salinity 32 Faktor salini NS Faktor trenin NS

19 Why is growth higher at lower salinity and training? Individual feed intake period 1 (BW g) g feed/day ppt 22 ppt 12 ppt Effect of salinity: p feeding days Cumulative feed intake g per tank period sampling 250 g sampling 450 g Effect of exercise: p 0.05 training no training feeding days sampling 250 g sampling 450 g feeding days feeding days

20 Survival, when the postsmolts were kept at 12, 22, or 32, through the entire trial Sensitive period % Cummulative survival Mean survival in cages 12 + Exercise 22 + Exercise 32 + Exercise 12 - Exercise 22 - Exercise 32 - Exercise salt, 99% 22 salt, 97% 32 salt, 71% Fish weight (g)

21 Survival when transfer from 0 ppt (start) or 12/22/32 to 32 at different sizes % Survival Transfer at 70 g (start) Training g 250g 450g g No training % Survival Transfer at 250 g no training training ppt 12 ppt 22 ppt 32 ppt 12 ppt 22 ppt 32 ppt Salinity before transfer to 32 Salinity before transfer to 32 At end of trial ~800 g % Survival Transfer at 450 g no training training ppt 22 ppt 32 ppt Salinity before transfer to 32 % Survival Change to 32 directly in existing12 tanks ppt 22 ppt 32 ppt Salinity before transfer to 32 All groups pooled, transferred to 12 new tanks, 32 FT % Survival All groups pooled

22 Summary survival, when the fish is transferred to 32 at different sizes 70g 250g 450g 800g

23 Sea water tolerance when transferred to 32-34, for 72 hours, at different sizes mg/l Plasma chloride 250 g no training 450 g training ~800 g ppt 22 ppt 32 ppt 12 ppt 22 ppt 32 ppt 12 ppt 22 ppt 32 ppt No significant effects of treatment (previous salinity or exercise)

exercised fish at 22 and 32, versus nonexercised, but this was not found at 12 NS NS Photo: Hoyle I, et al.

24 Welfare External welfare indcators: In our experience a set of sensitive indicators (e.g. density) Small, inconsistent effects on fin erosions Earlier manifestation of cataracts in fish kept at 32 RAS, versus 12 and 22 More external skin damage in exercised fish at 22 and 32, versus nonexercised, but this was not found at 12 NS NS Photo: Hoyle I, et al. (2007): A validated macroscopic key to assess fin damage in farmed rainbow trout (Oncorhynchus mykiss). Aquaculture 270, Cataract score (0-2) ppt 22 ppt 32 ppt Skin lesions (0-2) g 450 g 800 g 260 g 450 g 800 g

25 Skin health Important organ for pathogen and parasite control Les mucous and more tissue damage, at increasing salinity, and size Exercise had a negative effect on skin health at 22 and 32, but not at 12 Up-regulation of the stress related genes HSP70 and inos at 32,22 vs 12 We are establishing a skin analytical pipeline, since skin-health may be a particular problem Takle et al., unpubl

Commenced in July at Grieg SeaFood, Finnmark,

Oct. at Smøla Klekkeri and Settefisk, and in 21

26 Industry-scale testing in OPP Testing combination-line, closed-containment RAS Commenced in July at Grieg SeaFood, Finnmark, northern Norway, 12 and 22 salinity Foto: Nofima Foto: Nofima Testing combination-line, semi-closed in sea Foto: Nofima Start testing in Oct. at Smøla Klekkeri and Settefisk, and in m 3 tank, Marine Harvest, western Norway. Foto: Aquafarm Equipment

27 Conclusions Also salmon benefits from less salt and more exercise Closed-containment systems make environmental control possible Lower salinity: increased feed intake, growth rate, improved skin health Combination with exercise maximized this effect No general maturation was observed, via GSI Lower salinity improved removal efficency of TAN and CO2 Such types of findings may contribute to reduced costs in closedcontainment systems Production in closed-containment to 1 kg may contribute to smaller losses in Norwegian production Best group, 12 + exercise = 99% survival -6% at transfer to 32, = 93% survival to ~900 g BUT: at g the fish was very sensitive to handling This trial suggest a use for closed-contaiment RAS, also in grow-out phase in Norwegian aquaculture

Post-smolt welfare, performance and water quality in commercial scale, floating, semi-closed containment systems: Preline, Neptune and Eco-Cage.

Post-smolt welfare, performance and water quality in commercial scale, floating, semi-closed containment systems: Preline, Neptune and Eco-Cage. Sigurd Handeland Senior scientist, prof. II Production of