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larvi 2013 ghent university, belgium, 2-5 september 2013 6th fish & shellfish larviculture symposium Microparticulate enrichment of rotifers with taurine and other water-soluble substances and their subsequent effects on Lepidopsetta polyxystra larvae Matt Hawkyard

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Page 1: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

larvi 2013

ghent university, belgium, 2-5 september 2013

6th fish & shellfish larviculture symposium

Microparticulate enrichment of rotiferswith taurine and other water-soluble

substances and their subsequent effectson Lepidopsetta polyxystra larvae

Matt Hawkyard

Page 2: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Microparticulate enrichment of rotifers withwater-soluble substances and their

subsequent effects on Northern rock sole(Lepidopsetta polyxystra) larvae.

Matt Hawkyard, Ben Laurel,Yoav Barr and Chris Langdon

Page 3: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Live prey enrichment

Page 4: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Rotifers(cultured live prey)

Copepods(natural prey)

Page 5: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Rotifers Wild Zooplankton NRC

Taurine 0 7.7 -

Selenium <0.01 1.5 0.25-0.3

Iodine 3-8 50-350 0.6-1.1

Mæhre et al. 2012; Hamre et al 2008

Taurine in g kg-1 DWIodine and selenium in mg kg-1 DW

Water-soluble nutrients in rotifers vs.zooplankton

Page 6: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Problem: Water-soluble nutrients aredifficult to deliver to marine organisms.

Page 7: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Dissolved (Aqueous) vs. Particulateenrichments

Dissolvednutrient(s)

Microparticles or emulsionscontaining nutrient(s)

Page 8: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Phospholipidouter-

membrane

Aqueous core

LiposomesWax matrix

(yellow)

Aqueous orcrystalline core

Wax spray beads (WSB)Phospholipidouter-

membrane

Aqueous core

LiposomesWax matrix

(yellow)

Aqueous orcrystalline core

Wax spray beads (WSB)Phospholipidouter-

membrane

Aqueous core

Liposomes

Buchal and Langdon 1998

Page 9: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Phospholipidouter-

membrane

Aqueous core

LiposomesWax matrix

(yellow)

Aqueous orcrystalline core

Wax spray beads (WSB)Phospholipidouter-

membrane

Aqueous core

LiposomesWax matrix

(yellow)

Aqueous orcrystalline core

Wax spray beads (WSB)Phospholipidouter-

membrane

Aqueous core

Liposomes

Barr and Helland 2007

Page 10: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Wax Spray Beads (WSB) vs. aqueousmethods

Page 11: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Liposome enrichment

Water-solublemarker inaqueous corematerial

Lipid-solublemarker inphospholipidmembrane

Page 12: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Larval growth trials

Objective: Evaluate effects ofmicroparticle-WSB enriched rotifers on thegrowth of Northern rock sole larvae

Trial 1: Taurine-WSB

Trial 2: Taurine-liposome

Page 13: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Northern rock sole (Lepidopsetta polyxystra)

Egg Larva

JuvenileAdult

Page 14: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Northern rock sole distribution

Image: Fishbase.org

Hatfield Marine ScienceCenter, Newport, Oregon

Page 15: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Taurine-microparticle feeding trial

Trials occurred during rotiferphase (≈6 weeks)

Larvae grown in 100 L tanks

Flow though system ≈10° C, 35ppt seawater

Page 16: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Taurine

1. Low concentrations in rotifers

2. Sulfur amino acid

3. Not used in protein synthesis

1. Used for bile salt production,osmoregulation and neuralfunctions

Page 17: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Liposome equivalent

WSBequivalent

Zooplankton

Taurine enriched rotifers

Page 18: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Taurine-WSB feeding trial

Page 19: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Taurine concentrations in enriched rotifers

Note: Rotifers in all treatments were enriched with microparticles (empty or taurine filled)

Equal taurine providedto rotifers

Equal taurine providedto fish larvae

Control Dissolved50 mgtaurine l-1

Taurine-WSB(50 mgtaurine l-1)

Dissolved 4 gtaurine l-1

Page 20: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Larval growth (DW) during WSB trial

Page 21: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes
Page 22: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Taurine-liposome feeding trial

Page 23: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Equal taurine providedto rotifers

Equal taurine providedto fish larvae

Control Dissolved250 mgtaurine l-1

Taurine-Liposomes(250 mgtaurine l-1)

Dissolved15 gtaurine l-1

Taurine concentrations in enriched rotifers

Page 24: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Larval growth (DW) during taurine-liposome trial

Page 25: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Summary

• Microparticles were more efficient for enrichingrotifers with water soluble substances thanimmersion techniques

• Larval rock sole grew better when rotifers wereenriched with elevated concentrations of taurine

• Rock sole fed microparticle-enriched rotifersshowed intermediate growth response whencompared to the aqueous-taurine enrichedrotifers

Page 26: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Benefits of microparticles

• Reduced costs:– 92% to 95% reduction in materials costs– Reduced time to weaning

• Potential benefits:– Reduction of bacteria in rotifers– Delivery of additional nutrients or

antibiotics/vaccines

Page 27: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

NOAA NationalMarineAquacultureInitiative

USDA: Agricultureand food researchinitiative (AFRI):Competitive grant#2012-67015-19454

Acknowledgements

West CoastAssociation ofMarine Labs:Travel Award

Hatfield StudentOrganization:Travel Award

Page 28: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Questions?

Page 29: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

• Rotifers in all treatments were fed microparticles(empty or taurine containing)

• ½ of the rotifers fed to fish were fully enriched withAlgamac (taurine concentrations are reported forexperimentally enriched rotifers only, i.e. dietarytaurine concentrations are ½ of those reported

• There was no difference in the growth rates in fishfed 50% Algamac/50% microparticle enrichedrotifers (Controls) when compared to those fed100% Algamac

Lipids

Page 30: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes

Taurine leakage from WSB

Page 31: PowerPoint Presentation · 2013. 9. 17. · Aqueous core Liposomes Wax matrix (yellow) Aqueous or crystalline core Phospholipid Wax spray beads (WSB) outer-membrane Aqueous core Liposomes
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Cost (million rotifers-1)