FEEDING ZEBRAFISH:! A LESSON FROM AQUACULTURE CHRISTIAN LAWRENCE

Transcription

1 FEEDING ZEBRAFISH:! A LESSON FROM AQUACULTURE CHRISTIAN LAWRENCE

2 The Traditional! Zebrafish Feeding Paradigm! Westerfield, 1990, Nusslein-Volhard & Dahm, 2002 Paramecium, Artemia, Tetramin Flake, Powdered/Granular feed Larval diet, Artemia, Tetramin Flake Multiple times (2-6X) per day, regardless of life stage or application No published performance standards for this It works, but

3 Major proportion of resources allocated towards feed/feeding Performance is tightly correlated with diet and how it is delivered Improving performance has synergistic, positive effect on the science The importance of nutrition as experimental variable is underappreciated Why break with tradition?!

4 Refine methodology based on principles from commercial aquaculture (with a nod towards lab animal) FAO Aquaculture photo library

5 GOAL: A stable, defined diet that supports high performance

6 More Amount per feeding Flow rate Size of particle Protein content Frequency Less Larva Age of Fish Adult 24

7 BCH Feeding Paradigm! FEEDING Rotifers, CONTINUOUS Rotifers, 3-4X daily Gemma Micro 300, 1X 3-5% body weight Best et al., 2010! Lawrence et al., 2015! Lawrence et al., 2012! LIVE DIET! STATIC FLOW PROCESSED DIET! Day post-fertilization Parichy 2009

8 First Feeding Stage! (5-9dpf)! Small, digestible, nutritious meals on demand! Should be in the water column! Maximize energy savings/transfer! Keep water quality stable!

9

10

11

12 DAY 5! larva!

13 Production of million rotifers/day (scalable)

14 How can we recapitulate this in a laboratory setting? Fishconsult.org

15

16 DAY 5! Larvae! (rotifer densities > 500/mL)

17

18

19 Larval-Pre-adult Stage! (10-30dpf)! Small, digestible, nutritious meals frequently! Maximize energy savings/transfer! MORE BIOMASS! Most rapid phase of growth!

20 Producing million rotifers per day; only a small fraction of which being used to inoculate and drive early growth. What to do with the excess? Replace Artemia.

21 Artemia (AKA the third rail )! Expensive! Variable! Labor intensive! Waste producing!

22 Rotifers 2X Daily! 40 fish/l! 48dpf! Artemia 2X Daily! 40 fish/l! 48dpf!

23 Can we get enough biomass in with rotifers to meet performance goals?!

24 Spawning Performance! week 9 (60DPF)! Treatment Group Spawn Success Total Embryos Spawned Mean Clutch Size(MCS) MCS/female casper-rots wild-type-rots casper-art wild-type-art

25 Juvenile to Adult Stage! ( dpf)! 3-10%? body weight per day, multiple feedings! Maximize energy savings/transfer! High protein, proper PUFA/HUFA ratios, vitamins, minerals!

26 What should fish look like when they are ready for GM300? NOT READY READY When > 75% of the fish in a tank look READY begin feeding it!

27 Efficiency (especially at scale) is key.! What to feed! How often to feed it!

28 75% vs. 7% H 2 O content!! Artemia/rotifers are only efficient up until sub-adult stage Slide courtesy of Nick King

29 What feeding frequency is optimal for post-larval zebrafish?!

30 Gemma micro Nutrition -algae -predigested protein - natural fish meal blend (attractability) - conc. vit/min premix - phospholipids -conc. DHA oils -protein/fat ratio Microbiology/ Health -algae - Patented blend of organic acids & plant extracts -immunostimulants Physics/ Chemistry -particle shape -hygroscopy -leaching/binding -buoyancy -density -tensio-activity All natural ingredients : no colorants, GMO free! Slide courtesy of Nick King

31

32 One feed per day at 5% body weight/day is sufficient!

33 FEEDING Rotifers, CONTINUOUS Rotifers, 3-4X daily Gemma Micro 300, 1X 3-5% body weight LIVE DIET! STATIC FLOW PROCESSED DIET! Day post-fertilization Can we shrink this period down?! Can we refine approach for adult fish?!

34 Should feeding rates change once animals reach growth plateau?

35 Do we change profile of the diet? Do we simply adjust feeding rates? What about the application?

36 One Feed Per Day One Feed Every Other Day One Feed Every 5 Days High Usage Low Usage High Usage Low Usage High Usage Low Usage x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 x 10 Condition? Reproductive function? Histological changes?

37 Modern zebrafish research requires a more sophisticated approach to feeding and nutrition. We can borrow/adapt from aquaculture with an eye on laboratory animal science This will drive positive changes in efficiency, production, and in reproducibility of the science. Conclusions!

38 Acknowledgements EZM Organizing Committee Peter Alestrom ARP Jason Best, Althea James, Kara Maloney, Shane Hurley, Chris Lapointe, Likun Zhang BCH Zon Lab, Harris Lab Isaac Adatto, Nick King