Factors influencingFactors influencing skeletal malformations i A t li lt din Australian cultured

marine finfish

Jennifer Cobcroft Barbara NowakMalcolm BrownStephen BattagleneStephen Battaglene

TAFI is a joint venture between the State Government and the University of Tasmania

Outline

• Australian marine fish aquaculture industry

• Survey of hatcheries

• Species malformation case studiesp

• Future research direction

Australian Aquaculture

• Total finfish industry value 2007-08 AU$546 million; 41 200 t *AU$546 million; 41,200 t *

Salmonids Southern Bluefin Tuna

* ABARE, 2009

AU$299 million; >25,500 t AU$187 million; 9,800 t

Australian Aquaculture

• Marine fish value (based on hatchery produced fry)~AU$57 million; ~ 5 600 t *~AU$57 million; ~ 5,600 t

• Major speciesBarramundi (Lates calcarifer)Barramundi (Lates calcarifer) AU$34 million; 3,360 t

Yellowtail kingfish (Seriola lalandi)

Mulloway (Argyrosomus japonicus)

Australian bass (Maquaria novemaculeata)

* ABARE, 2009

3 spp combined < AU$23 million; 2,240 t

Australian Aquaculture• Species under development

Flowery grouper (Epinephelus fuscoguttatus)• Flowery grouper (Epinephelus fuscoguttatus)

• Gold Spot grouper (Epinephelus coioides)

• Cobia (Rachycentron canadum)• Cobia (Rachycentron canadum)

• Striped trumpeter (Latris lineata)

• Australian snapper (Pagrus auratus)pp ( g )

• Southern bluefin tuna (Thunnus maccoyii)

Images courtesy of L. Woolley and Clean Seas Tuna

Australian AquacultureTarget to increase finfish aquaculture

production from 41,000 toproduction from 41,000 to 100,000 tonnes by 2015 *

2 mm

Provision of sufficient numbers of high quality fry is a bottleneck to industry expansion

• What are the issues at the hatchery level?

* Hone, 2008 (FRDC Fish 16(2), 14-17)

Hatchery Survey

• Survey emailed to 26 marine fish hatcheriesF ll ith t l h i t i• Follow-up with telephone interview

• incidence and severity of skeletal malformations

• perceived impact on hatchery production

• techniques used to rear larvae

• Hatchery informationkept confidential

Hatchery Survey• Responses from 18 hatcheries (69%)

• Samples from 2 ‘no issue’ hatcheries had >5% severe malformations (i e 56% hatcheries with an issue)

* Minimal production 2008-2009

malformations (i.e. 56% hatcheries with an issue)– more work to be done

Barramundi• Spinal Normal Short body

*

LordosisVertebral body – compression, dorsal or ventral shift in centre*

Barramundi• Jaw - occasional

Images providedImages provided by Darwin

Aquaculture CentreCentre

Solved by changing th i l lthe commercial algae pastes for rotifer and

larval culture *

Normal jaw Subtle deformedjaw

Medium deformedjaw

* Bosmans personal communication; Schipp et al 2007

Barramundi• Jaw - occasional

‘Pinched’ lower *Normal

* Similar to the description of Fraser and de Nys, 2005

Grouper• Epinephelus fuscoguttatus and Epinephelus coioides

• Jaw (maxilla, dentary) and some spinal

Day 24. E. fus. Day 40. E. fus. Day 40. E. fus.

D 32 E fDay 40. E. fus. normal Day 40. E. fus.

Day 32. E. fus.

Day 42. E. coi.

Yellowtail kingfish• Jaw – several malformation types

Yellowtail kingfish• Jaw – several malformation types

Yellowtail kingfish• Jaw – several malformation types

Yellowtail kingfish

• Jaw – onset of fusion from day 11

Maxilla located inside lower jawjDay 11

0.5 mm 0 5 mm0.5 mm

0.5 mm

0.5 mm

Yellowtail kingfish

• Jaw – onset of fusion, force exerted on maxilla and premaxilla by lower jawmaxilla and premaxilla by lower jaw movement

Striped trumpeter• Occasional spinal

Striped trumpeter

Day 44 1Jaw

0Jaw

malformation index

2

0.5

3

Striped trumpeterOnset of jaw malformation

Jaw score

80

10000.512

Jaw scoreco

re (%

)

60

23

Jaw

s

20

40

27 30 35 41 50 57 63 70 79 90 1000

Age (days post-hatching)

Striped trumpeter

• Conducted a series of experiments (di t d i t l f t )(dietary and environmental factors)

• Experimental system – 24 x 300 L tanksC t ll d h t i d t t• Controlled photoperiod, temperature

• Flow through, recirculation or both• Ozonated seawater

Striped trumpeter

• Live feed - Artemia enrichment with Vitamin C– Larvae fed Artemia Day 17 to Day 44– Multifactor experiment (Vit C and prey density)

Artemia with Vitamin C enrichment (ascorbic acid– Artemia with Vitamin C enrichment (ascorbic acid, aa, 7,900 µg g-1) and without (aa 640 µg g-1)

– 4 Artemia densities (0.125, 0.25, 0.5 and 1.0 mL-1), ( , , ),fed 4x daily

Battaglene et al, unpublished data

Striped trumpeter

• Live feed - Artemia enrichment with Vitamin CDay 44Day 44

) 80

100 Lower incidence with Vitamin C (8%) enrichment than

scor

e (%

)

40

60without (11%)

Jaw

20

40

1-NI

L0.

5-NI

L0.

25-N

IL0.

125-

NIL

1-VI

TC0.

5-VI

TC0.

25-V

ITC

125-

VITC

0

Battaglene et al, unpublished data

Treatment diet

0 0 0 0.1

Striped trumpeter

• Tank colour, enrichment & water type– Tank colour (black & white)– Diet (enriched & non-enriched)– Water type (green & clear)ate type (g ee & c ea )

• Larvae cultured Day 1 to Day 29• Rotifers Day 6 – Day 19• Artemia Day 18 – Day 29• Assessed growth, survival, jaw malformation• Daily assessment of larval walling behaviour

Cobcroft et al, unpublished data

Striped trumpeter

Away from wall

Walling behaviourWalling behaviour

Results – jaw malformationA) Day 15

80

100

0 0 5

B) Day 20

80

1007.5 mm when malformation first observed

Jaw

sco

re (%

)

40

60

0.5 1 2 3

40

60Day 29In clearwater treatments

first observed

0

20

C) Day 24100

D) Day 29100

0

20 treatments WH CL AM 70 ± 15%> all others

scor

e (%

)

40

60

80

40

60

80In black tanksCW > GWAM > NE

CL NE

CL AM

CL NE

CL AM

R NE

R AM

Jaw

0

20

40

CL NE

CL AM

CL NE

CL AM

GR NE

R AM

0

20

40 AM > NE

Treatment

WH C

LW

H CL

BL CL

BL CL

BL GR

BL GR

Treatment

WH C

LW

H CL

BL CL

BL CL

BL GR

BL GR

Cobcroft et al, unpublished data

Results – walling behaviour

100

g (%

) 80

WH CL NEWH CL AMBL CL NEBL CL AMBL GR NE

on w

allin

40

60BL GR NEBL GR AM

Prop

orti

20

10 15 20 25 30

0

Age (days)Mean + SD

Cobcroft et al, unpublished data

Results j lf ti lli- jaw malformation vs walling

Correlation between jaw malformation and wallingj gs

(%) 100 AM

r2 = 0.830P < 0.001

orm

ed ja

ws

60

80

with

mal

fo

40

0 10 20 30 40 50 60 70

Larv

ae w

0

20 NE - nsr2 = 0.197P = 0.148

Proportion walling (%)0 10 20 30 40 50 60 70

Cobcroft et al, unpublished data

Comparison with tank colour

Tank colour• Design: 6 tank

colours, 4 reps• Larvae culturedLarvae cultured

Day 16 to Day 44• Daily assessment

of larval wallingof larval walling

Cobcroft & Battaglene, 2009 (Aquaculture 289: 274-282)

Comparison with tank colour

100 black blue

ling

(%)

60

80

blue green marble red white

Walling behaviour

port

ion

wal

l

40

60Pr

op

0

20

Age (days)20 25 30 35 40 45

Mean ± SD

Cobcroft & Battaglene, 2009 (Aquaculture 289: 274-282)

Comparison with tank colour100

0 0 5

Jaw score

re (%

)60

80 0.5 1 2 3

Jaw malformation

Jaw

sco

r

40Day 44

k e e n d e

0

20

Tank colour

black

marble blu

e

gree

n

red

white

Cobcroft & Battaglene, Aquaculture 2009

0 Correlation between jaw malformation and walling

Day 44aw

s (%

)

60

80

Day 442 0 871

alfo

rmed

ja

40

60 r2 = 0.871P < 0.001

3

ae w

ith m

a

20

1 mm0 10 20 30 40 50 60

Larv

0

Proportion walling (%)

Cobcroft & Battaglene, 2009 (Aquaculture 289: 274-282)

Striped trumpeter• Improvement in quality of larvae from production trials• 75% fish with normal & minor malformations at75% fish with normal & minor malformations at

Day 100 in 2 trials in 2006 & 2007

2005STT1 05

2007STT2 07 Tank 2

80

10000.512

STT1-05

80

10000.512

STT2-07 Tank 2

aw s

core

(%)

40

60

23

aw s

core

(%)

40

60

23

25 28 30 34 39 45 50 60 70 80 90 100

126

153

170

187

197

200

Ja

0

20

20 23 25 30 36 37 40 50 60 71 80 91 100

Ja

0

20

1 1 1 1 1 1 2 1

Striped trumpeter

• Live feed - rotifer enrichment with Vitamin A (retinol palmitate)Vitamin A (retinol palmitate)– Larvae stocked at day 1

R tif i h d ith 8 diff t ti l– Rotifers enriched with 8 different retinol palmitate levels (enrichments contained: 0, 100, 300, 750, 1500, 3000, 5000, 10000 mg L-1; NutraKol)

– Rotifers fed day 6 to 18Artemia (all AlgaMac 3050 enriched) fed day– Artemia (all AlgaMac 3050 enriched) fed day 17 to 43

Negm et al, unpublished data

Striped trumpeter

Slightly higher atJaw score

Day 43

Slightly higher at lower VA doses 100 and 300 mg L-1.

Overall, jaw malformation higher than tank colour experimentexperiment.

Analysis of live feed VA profile and other

lf timalformations are underway.

Negm et al, unpublished data

Future Research

• Striped trumpeter (ARC Linkage) - vitamin A in live feeds on malformation and gene expression and tank systemon malformation and gene expression and tank system design

• Yellowtail kingfish (Seafood CRC)• Southern bluefin tuna (Seafood CRC, CST)• Communication with industry & identification of target

( b di)areas (e.g. barramundi)• International collaboration

5 mm

AcknowledgementsTAFI – MRL Research team StaffStephen Battaglene

PhD CandidateReham Negmp g

Alan BeechJenny CobcroftMelanie EvansRoss Goldsmid

g

Tanaz JungalwallaTom Litjens Anna Overweter Gavin ShawBill Wilkinson

Acknowledgements

Hatchery and Research staff of Clean Seas Tuna

D i A lt C t NT DRDPIFRDarwin Aquaculture Centre – NT DRDPIFRNorthern Fisheries Centre – QDPI

Huon Aquaculture CompanyNutraKolSkrettinggClean Seas TunaTasmanian Government