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STRANDBÚNAÐUR 2018 Grand Hótel Reykjavík, 19. – 20. mars 2018

Renée Katrin Bechmann

Environmental effects of chemicals used against salmon lice

21 March 2018 In aquaculture, pesticides are used against parasitic salmon lice to protect the health of farmed and wild Atlantic salmon.

- How can this use of pesticides as medicine affect our coastal marine environment?

Economic consequences  Costs billions for the aquaculture industry Salmon lice and the war against lice Environmental concequences  Animal welfare for farmed and wild salmon How to get rid of lice  Risk for reduced stocks of wild salmon  Kill them with chemicals  Animal welfare and overfishing of cleaner fish  Use cleaner fish and other non-chemical methods  Risk for non-target crustaceans, and the rest of  Protect the salmon from lice in (semi-)closed cages the coastal ecosystem The perfect anti-salmon lice medicine

Low toxicity to:

Humans High toxicity to: Salmon lice Salmon = target crustaceans Must eat the chemical or swim in a solution

 Fast depuration after treatment of the fish The environment including non-target crustaceans Must die!  Fast degradation  Low bioavailability … and not develop resistance  Low toxicity kg used per year (log scale) 100000000 10000000 1000000 100000 10000 1000 100 10 1

1981 againstChemicals used 1981lice salmon 1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 -

2010 2017 2011 2012 2013 2014 2015 2016 2017 peroxide Hydrogen benzoate Emamectin Teflubenzuron Azamethiphos Effects of Releeze medicine feed containing diflubenzuron High mortality of shrimp larvae exposed to tiny particles of medicated feed with diflubenzuron (DFB) as the active ingredient

100 Ambient Climate Control 80 Future Climate Control 2 g 60 3.5 mm pellets * * 40 DFB at Flow: 150 ml/min Ambient Climate *

20

% Survival of larvae of shrimp % Survival DFB at Pandalus borealis Future Climate 0 0 5 10 15 20 25 30 14 d exposure

Age of larvae (days post hatch) Ambient climate: pH 8.0, 7.0 °C Future Climate: pH 7.6, 9.5 °C Mortality High mortality of adult shrimp eating a few pellets Releeze 7.0°C pH 8.0

* *

9.5°C pH 7.6 • Control shrimp moulted successfully • Hardly any shrimp exposed to medicine feed for two weeks * managed to moult, and most died during moulting * • 4 pellets Releeze (ca 0.1 gram) is enough to kill adult shrimp that need to moult during the next 2-3 weeks • 8 million kg of this medicine feed was used in 2016

Female shrimp were fed Releeze a few times before moulting Mean for 6 replicate tanks with 17 shrimps in each - Are flubenzurones (from feed) bioavailable for crustaceans in the field? - Yes

 Langford et al. 2014, NIVA: Diflubenzuron and teflubenzuron were detected in shrimp, crab and blue mussels sampled 3 weeks after treatment.

 Samuelsen et al. 2015, IMR: Teflubenzuron was still detected in crustaceans and polychaets 8 months after treatment.

 Samuelsen et al. 2014 & 2015, IMR: The results indicate that the concentrations of teflubenzuron in king crab, shrimp, squat lobster and Norway lobster were high enough to induce mortality if moulting was imminent. Effects of «bath chemicals»

Hydrogen peroxide – in Paramove Strong oxidizing agent, non-specific

Azamethiphos – in Salmosan Neurotoxic acetylcholinesterase inhibitor

….. leading to paralysis

Deltamethrin – in AlphaMax Neurotoxic … leading to paralysis E xperiment with shrimp ( Pandalus borealis)

AlphaMax Salmosan AlphaMax Salmosan Paramove

DEL AZA DEL AZA H2O2

Deltamethrin Azamethiphos Deltamethrin Azamethiphos Hydrogen peroxide

5 L 5 L 5 L 5 L 5 L stock solution stock solution stock solution stock solution stock solution

Peristaltic pump Peristaltic pump Peristaltic pump Peristaltic pump Peristaltic pump 3.2 ml/min with 6 pump heads with 6 pump heads with 6 pump heads with 6 pump heads with 6 pump heads

10 shrimp with eggs 10 shrimp with eggs 10 shrimp with eggs 10 shrimp with eggs 10 shrimp with eggs Seawater Seawater Seawater Seawater Seawater 750 ml/min 750 ml/min 750 ml/min 750 ml/min 750 ml/min

tanks X 6 tanks X 6 tanks X 6 tanks X 6 tanks X 6

Control DEL AZA AZA + DEL H2O2 Sceening tests with AlphaMax (deltamethrin)

Control Control 100% 100%

50% 50%

0% 0%

2 hours @ 10 times diluted AlphaMax solution + 2 hours recovery 8 hours @ 1000 times diluted AlphaMax solution acc. Dead 100% 100% Lying down

50% R. I. P. Swimming 50%

Standing 0% 0% 01:15 01:30 01:45 02:00 01:15 01:30 01:45 02:00 15 min 30 min 45 min 60 min 15 min 30 min 45 min 60 min 2 hours exposure 2 hours recovery Main experiment, adult shrimp, means, n = 6 2 ng/L deltamethrin Control 1000 times diluted salmon treatment concentration of 100% AlphaMax 100%

50% 50%

0% 0%

0.1 µg/L azamethiphos + 2 ng/L deltamethrin 0.1 µg/L azamethiphos 1000 times diluted of the salmon treatment concentrations of Salmosan and AlphaMax 1000 times diluted salmon treatment concentration of 100% Salmosan 100%

50% 50%

0% 0% 1d 2d 3d 4d 5d 6d 7d 8d 9d 1d 2d 3d 4d 5d 6d 7d 8d 9d pre pre pre pre pre pre pre pre pre pre pre pre pre pre 10d 11d 12d 13d 10d 11d 12d 13d post post post post post post post post post post post post post post 1. 2. 3. 4. 5. 6. 7. Recovery 1. 2. 3. 4. 5. 6. 7. Recovery pulse pulse pulse pulse pulse pulse pulse pulse pulse pulse pulse pulse pulse pulse

1 dead shrimp after 7 pulses 2 h exposure per day 2 weeks recovery 1000 times diluted AlphaMax solution for 7 days 1,5 caused reduced feeding for adult shrimp

1,0 Control

+ SD) 0,5

0,0 mean ( 1,5 day

shrimp 0.1 µg/L azamethiphos 1,0 per

0,5 1000 times diluted salmon treatment concentration of Salmosan shrimp

0,0 per 1,5 2 ng/L deltamethrin (AlphaMax) rate for rate for 1,0 consumed 0,5 1000 times diluted salmon treatment concentration of AlphaMax

pellets 0,0

of 1,5

1,0 Feeding 1000 times diluted mix of Salmosan and AlphaMax Number

0,5 n = 6 replicate tanks per treatment, 0,0 each with 10 shrimp at start Day 1-2 Day 3-4 Day 5-6 Day 7-8 Day Day 9-10 Day Day 11-12 Day 13-14 Day 15-16 Day 17-18 Day 19-20 Day Histological assessment of adult shrimp – Digestive gland alterations (work in progress)

7 x 2 hours exposure to 1000 times diluted treatment water of Salmosan and/or AlphaMax

Salmosan + Control Salmosan AlphaMax AlphaMax

Ec Rc Rc Rc Bc Rc ABL

Fc Bc Abnormal lumen (ABL) and hemocytic infiltration (HI) in the interstitial sinus HI (IS) were observed.

Preliminary conclusion:

Both Salmosan and AlphaMax caused tissue damage, and it was worse in the combined exposure DIRECT EXPOSURE OF SHRIMP LARVAE

High mortality of larvae exposed for 2 hours to 1000 times diluted AlphaMax or AlphaMax + Salmosan

100 Salmosan 80 * Control 2 hour exposure 60 AZA + two weeks recovery

40 AlphaMax DEL

% Survivors * AZA/DEL 20 AlphaMax + Salmosan * * 0 0 2 4 6 8 10 12 14

Age of larvae (days) DIRECT EXPOSURE OF SHRIMP LARVAE

Low swimming activity of larvae exposed for 2 hours to 1000 times diluted AlphaMax or AlphaMax + Salmosan

Day 4 2000 2 hours exposure

hour + two weeks recovery 1600

1200

800 Low swimming beambreaks per per beambreaks

400

Mean * *

0 Control AZA DEL DEL/AZA

Salmosan AlphaMax AlphaMax Salmosan - How do shrimp respond to pulses of diluted Paramove (H2O 2) treatment water? High mortality of adult shrimp exposed to diluted Paramove (H2O2) Dead Immobilized Swimming 100% Increased mortality after exposure to Standing 3 pulses of 1000 times diluted

50% salmon treatment concentration

0% Control Control Control Control Control Control Control 1.5 mg/L 1.5 mg/L 1.5 mg/L 1.5 mg/L 1.5 mg/L 1.5 mg/L 1.5 mg/L Before After Before After Before After 1 d 100% Very high mortality after exposure to 3 pulses of 100 times diluted salmon treatment concentration

50% Mortality also increased 3 days after 1 pulse exposure: 0% Delayed effects! Control Control Control Control Control Control Control Control Control 15 mg/L 15 mg/L 15 mg/L 15 mg/L 15 mg/L 15 mg/L 15 mg/L 15 mg/L 15 mg/L Before After Before After Before After 1 d 1 d 2 d

3 x 2 h exposure pulses Recovery Significantly reduced feeding rate for shrimp during and after exposure to

3 pulses of 15 mg/L and 1.5 mg/L H2O2

1 pulse = 2 hours 100 times diluted 1000 times diluted Increased swimming activity during exposure to 15 mg/L H2O2 100 times diluted salmon treatment concentration of Paramove Indications of increased swimming activity for adult shrimp also after exposure to 1000 times diluted treatment concentrationof Paramove

H2O2 Mean for 4 shrimp

Activity of shrimp monitored continuously over seven days One hour exposure to 1.5 mg/L and 15 mg/L H2O2 caused gill damage in adult shrimp

1000 times diluted treatment solution for salmon

100 times diluted peroxidative stress Significant tissue damage in the digestive glad of adult shrimp exposed to 1.5 mg/L and 15 mg/L H2O2 1000 times diluted treatment water of AlphaMax & Paramove, or a few pellets of Releeze medicine feed can kill shrimp

- Is dilution the solution when pesticides are used as medicine?

- Maybe not? Page et al. (2014), Fisheries and Oceans Canada: Within a couple of hours after release, bath chemicals may be advected up to 1-2 kilometers, and the concentration is estimated to be 100 – 1000 times diluted

Samuelsen et al. 2015, IMR: Particles of medicine feed and feces from fish eating medicine can be transported 10 times 100 times Bath more than 1 km away from the farm diluted 1000 diluted treatment times diluted 1-2 km Please think about What is acceptable?

 How important is it to protect the coastal marine

Aquaculture is important for Norway ecosystem? Are shrimp & co “expendable”?

In 2017 we had ca. 3400 cages at 550 locations  How important is it to protect wild Atlantic salmon …… x 5 in the future? from lice (and escaped farmed salmon)?

= Sustainable?  Should farmed salmon be protected from lice in closed cages to stop the use of chemical treatment? Thank you all for listening!

Thank you to the project participants: Maj Arnberg 1), Emily Lyng 1), Thank you to the Research Council of Norway, Stig Westerlund 1), Shaw Bamber 1), EU and Solvay for the funding: Sree Ramanand 1), Mark Berry 1), Elisa Ravagnan 1), Jannicke Moe2), Dag Ø. Hjermann2), Paul Seear3), 1. The RCN project FluClim (PL: Renée K. Bechmann) Piero Calosi4), Katherine Langford2) Alfhild Kringstad2), Thomas Rundberget2), 2. The EU project ECOAST (IRIS WP leader Thorleifur Agustsson) Alessio Gomiero1), Tjalling Jager5), Frederike Keitel-Gröner, Thorleifur Agustsson1), 3. The RCN project PestPuls (PL: Renée K. Bechmann) Les Burridge6), Renée K. Bechmann1)

4. The Solvay project (PL: Renée K. Bechmann) 1) IRIS 2) NIVA 3) University of Leicester 4) Université du Québec à Rimouski 5) DEBtox Research 6) Burridge Consulting Inc