Hardangerfjordseminaret 2014 Thon Hotel Sandven, Norheimsund, 21. november. Stemmer det?

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1 Hardangerfjordseminaret 2014 Thon Hotel Sandven, Norheimsund, 21. november Media skriver: Lakselusmedisin tar knekken på rekene Stemmer det? Resultater fra prosjektet FLUCLIM - Effects of diflubenzuron on Northern shrimps (Pandalus borealis) at ambient and future climate conditions IRIS Biomiljø Lab-team: Renée K. Bechmann, Shaw Bamber, Emily Lyng, Stig Westerlund, Sree Ramanand, Marianne Nilsen Elisa Ravagnan - Modelling NIVA Katherine Langford Analysis of Jannicke Moe & Dag Ø. Hjermann - Modelling University of Leicester Paul Seear Genomics Université du Québec à Rimouski Piero Calosi Scientific advisor Finansiert av Forskningsrådet Havet og Kysten ( )

2 Markus J. Thonhaugen nrk.no

3 T h e P r o b l e m.. T h e e n v i r n o m e n t a l C h a l l e n g e A m b i e n t c l i m a t e ph C F u t u r e c l i m a t e Ocean acidification (ph 7.6) and increased temperature (10 C) Shrimps Pandalus borealis Diflubezuron from medicated fish feed A chitin synthesis inhibitor Log K ow 3.8 Benzoylurea pesticide Anti-parasitic drug against salmon lice Controls: Normal fish feed

4 Shrimp larvae are exposed to from suspended particles + dissolved : Negative effects? H o w c a n s h r i m p s be e x p o s e d? Medicated fish feed 14 days treatment of salmon to get rid of salmon lice Positive effect for salmon: Salmon lice die during moulting Experiment done spring 2014 Pilot test spring 2014 Main experiment next year (Re)suspended particulates Dissolved from fish feed Adult shrimps eat feed waste, feces and sediment: Negative effects on non-target epibenthic crustacean? Feed waste Feces from salmon Sediment

5 Temperature controlled seawater Heat exchangers 10 C 7 C Continuos flow exposure system + CO 2 Aqua Medic Pressure regulator system Connection plug C CO 2 Solenoid valve Stock solutions of sediment slurry with medicated or clean fish feed

6 Main exposure: Medicated fish feed (0.66 g /kg) 3.5 mm pellets 2 g Flow: 200 ml/min A little extra : 8% fish feed in sediment: 0.5 mg/l Shrimp larvae were exposed to the pellets for 2 weeks from hatching

7 Tr e a t m e n t s & T i m e : Ambient Climate 7 C, ph 8.1 x 6 at Ambient Climate 7 C, ph 8.1 x 6 Future Climate 10 C, ph 7.6 x 6 at Future Climate 10 C, ph 7.6 x 6 Climate acclimation of shrimps (females with embryos) Hatching Age of larvae (days) Two weeks exposure to medicated pellets with

8 Ambient climate: 7 C and ph 8.1 Future climate: 10 C and ph 7.6 X 6 X 6 Exposure to X 6 X 6 Exposure to X 6 batches X 6 batches X 6 batches X 6 batches Control OA/T OA/T +

9 Analysis of by Katherine Langford, NIVA Total concentration of diflubenzuron in the aquaria (dissolved and on particles) 1 mm mesh 0.2 L/min (288 L in 24 h) After 4 hours: 2 µg/l 1.32 mg in 2 g pellets After 24 hours: 1 µg/l Conc. of 15 litre Fish feed after 24 h in the cylinder: 3.5 mm pellets

10 µg/l Analysis of by Katherine Langford, NIVA C o n c e n t r a t i o n of d i f l u b e n z u r o n a f t e r 4 h a n d 2 4 h 3,0 Total Concentration of in the aquaria (mean + SE) i.e. attached to feed particles and dissolved in the water 2,5 2,0 1,5 1,0 0,5 0,0 4 h 4 h 24 h 24 h n = 3 n = 3 n = 3 n = 3 Ambient Climate + Future Climate + Controls samples: < µg/l was analyzed at NIVA by liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS)

11 Percent survival of shrimp larvae SURVIVAL for shrimp larvae Mean percent survival for 6 replicate batches of shrimp larvae exposed to at two climate scenarios % Ambient Climate Control % Future Climate Control 20 0 E x p o s u r e t o D F B o r c l e a n p e l l e t s - 82 % Days post-hatch Ambient Climate + Future Climate +

12 Development time (zoeal progression) for shrimp larvae The tail of the shrimp larvae change when they develop from stage II III - IV Photo: Tandberg & Arnberg Stage I zoea Stage II zoea Stage III zoea Stage IV zoea Time Shrimp larvae go through five pelagic zoea stages and one megalopa stage before they settle as post-larva We followed them until stage 4

13 C o n t r o l C o n t r o l C o n t r o l C o n t r o l C o n t r o l C o n t r o l D F B C o n t r o l C o n t r o l L A R VA L D E V E L O P M E N T T I M E f o r the s u r v i v o r s 100 % stage II % stage III % stage IV Ambient Climate No stage II No stage III No stage IV 80 Future Climate Day 6 Day 9 Day 13 Day 17 Day 19 Day 27 Day no = age of shrimp larvae Exposure to medicated () or clean (control) pellets

14 1 litre Feeding rate for stage II and IV larvae 5 shrimp larvae and 150 Artemia nauplii in 1 litre seawater. Pandalus borealis larvae Wait 5 hours. Count remaining Artemia. Artemia nauplii

15 Number of Artemia eaten per shrimp larvae per hour Feeding rate for the survivors 6 Stage II zoea Stage IV zoea Control Number of Artemia eaten per shrimp larvae per hour Control Control Control No stage 4, these are stage 3! 0 Ambient 1 Climate 2 Future 3 Climate 4 Ambient 5 Climate 6 Future 7 Climate 8 code B

16 Shaw Bamber, IRIS B S T «B a m b e r s S w i m m i n g T e s t» L i g h t I R l i g h t Detector Shrimp larvae are photo-positive (they swim towards the light) - Will affect swimming activity at ambient or future climate conditions?

17 Shaw Bamber, IRIS S w i m m i n g a c t i v i t y Ambient Climate Similar swimming activity for control and exposed STAGE I and II larvae. Future Climate STAGE I: Similar swimming activity for control and exposed larvae. STAGE II: Lower swimming acitivity for exposed larvae than control larvae. STAGE I: Before first moult STAGE II: Those that survived first moult Control (future climate) (future climate) Example - Raw data

18 Conclusions from the shrimp larvae experiment medicated pellets caused high mortality of shrimp larvae and negative effects on the surviving larvae More mortality and more negative effects on the survivors at future climate - What about the adult shrimps? - Are they more tolerant to?

19 P i l o t e x p e r i m e n t Effect of exposure on post-hatch moulting Shrimps in large tanks and in 15 litre aquaria Exposed to a few grams of medicated or clean fish feed in the water for 2-4 weeks Upubliserte resultater fra HAVKYST prosjekt #234407/E40. Prosjektleder: Bechmann, IRIS Biomiljø.

20 P i l o t e x p e r i m e n t Control shrimps moulted successfully in the lab a few weeks after all eggs had hatched Shrimp exit

21 P i l o t e x p e r i m e n t

22 P i l o t e x p e r i m e n t S h r i m p s d i e d d u r i n g m o u l t i n g w h e n e x p o s e d t o m e d i c i n e p e l l e t s c o n t a i n i n g D F B ( d i f l u b e n z u r o n ) Flubenzurones are chitin synthesis inhibitors: they stop the organism from casting its exuvium during the molting process resulting in death. Exposed to a few grams of medicated fish feed in the water for 2-4 weeks

23 The Score Controls exposed Successfull moults: 14-0 Dead of unknown cause: 1-3 Dead during moulting: 0-13 vs

24 Oppsummering! Rekelarver og andre krepsdyr i planktonet kan få i seg dette Rekelarver er veldig følsomme for diflubenzuron fra medisin-fôr! Medisinfôr med flubenzuroner Fôr - spill Lite metaboliseres Alt skilles ut raskt Strømmen tar med seg rester av medisin-fôr, feces og forurenset sediment Feces med flubenzuroner Voksne reker kan spise dette og få problemer under skallskiftet Flubenzuroner har «lang holdbarhet» i det marine miljø

25 Konklusjon! Både rekelarver og voksne reker dør under skallskifte når de utsettes for medisin-fôr med diflubenzuron Mer negative effekter ved høyere temperatur (framtidig klimascenarie) Så svaret er: Ja! Lakselusmedisin kan ta knekken på rekene hvis for mye slippes ut i fjordene våre.

26 - So what? - Kan vi både beskytte ville krepsdyr og laksens helse?? - Bør vi bruke «føre-var-prinsippet» hvis vi er usikre på hvor mye flubenzuroner lokale krepsdyrpopulasjoner tåler? Eller bør vi forske mer? - Kan vi kun bruke andre metoder enn pesticider for å kvitte oss med lakselus. mens vi venter på lakselus-vaksinen til Frank Nilsen & co?. eller i hvert fall droppe pesticider med lang holdbarhet og høy toksisitet for ville krepsdyr? P l e a s e!

27 On- g o i n g p r o j e c t w o r k : Genomics The aim is to develop a genetic marker for exposure to chitin synthesis inhibitors in shrimps Paul Seear is working with the shrimp genomics F u t u r e ( ) p r o j e c t w o r k : Main experiment with adult shrimps F u t u r e ( ) p r o j e c t w o r k : Elisa Ravagnan Jannicke Moe & Dag Ø. Hjermann Modelling. analyse the multiple stressors impacts of increased temperature and OA together with exposure on Northern shrimp populations

28 T a k k t i l d e r e s o m h ø r t e p å! Thank you for the money: H A V K Y S T NFR Thak you for the science: The local crowd Partners Stig Westerlund Emily Lyng Shaw Bamber Elisa Ravagnan Marianne Nilsen Sreerekha S. Ramanand Renée K. Bechmann Katherine Langford, Jannicke Moe Dag Ø. Hjermann University of Leicester Paul Seear Université du Québec à Rimouski Piero Calosi M o r e i n f o : r k i r i s. n o Photo: Elisabeth Tønnessen