The future prospects for aquaculture breeding in Europe

Paris, October 2008

STERILIZING FISH TO PROTECT

BIODIVERSITY AND IMPROVE QUALITY

Alicia Felip

Institute of Aquaculture of Torre de la Sal (IATS)

Spanish Council for Scientific Research (CSIC)
Ribera de Cabanes s/n, 12595 Castelln, Spain
afelip@iats.csic.es
 The future prospects for aquaculture breeding in Europe
Paris, October 2008

OVERVIEW
Thorgaard, 1983
Scientific knowledge Purdom et al., 1983
Industrial application Ihssen et al., 1990
Practical knowledge gaps
Main reserach axes
Benfey, 1999
Hulata, 2001; Felip et al., 2001
EUROPEAN FARMED FISH Piferrer et al., 2007

Salmon and Rainbow trout

Tilapia
Common carp
Sea bass and Sea bream
Turbot, Sole and Halibut
Cod IDENTIFICATION OF
Meagre RESEARCH PRIORITIES
new species?? FOR THE INDUSTRY
Chromosome set manipulation in fish: Triploidization
Gamets

X Disruption of the second

meiotic division
Normal eggs
Sperm (1N)
(1N)

SHOCK Treatments

1N Pressure Chemical
2nd polar body shocks shocks
Sperm
(1N) (1N)
Thermal
shocks

3N
Variables

Timing
Intensity
Duration

Triploids (3n)

Directive 90/220/CEE of April 23 of 1990

 The future prospects for aquaculture breeding in Europe
Paris, October 2008
Scientific knowledge

Atlantic salmon
(Salmo salar)
Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Pressure 100 88.6 Juveniles 2n = 3n
Heat 100 92.5 Adults 3n > 2n
Commercial-scale : Canada

Rainbow trout
(Oncorhynchus mykiss)
Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Heat 100 57.7 Juveniles 2n = 3n
Adults 3n > 2n
Commercial-scale : Canada, France, Japan

9 Production of aneuploid sperm

9 Preference for 3n females
 The future prospects for aquaculture breeding in Europe

Production of all-female triploids Paris, October 2008

Both sexes Genetic sex determination: XX/XY

XX females / XY males
Gynogenesis approach
Sex-reversal approach
Gynogenesis
By fertilizing eggs with sperm from neomales
100% females XX Triploidy approach

Androgens

Genotipically females Genotipically females

Neomales XX X XX
Phenotipically males Normal fish

Triploidization

100% females XX
genotipically/phenotipically
and sterile
 The future prospects for aquaculture breeding in Europe
Paris, October 2008
Scientific knowledge

Sea bass
(Dicentrarchus labrax)
Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Pressure 100 71 Juveniles 2n = 3n
Cold 89-100 40-80 Adults 2n > 3n
Inicial tests for commercial-scale : France

Sea bream
(Sparus aurata)
Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Heat 100 40 Juveniles 2n = 3n

Questionable interest??

350-450 g market size (all-males); 600 g sex change to females

Depending on cycle production, sexual maturation can appear
before commercialization
 The future prospects for aquaculture breeding in Europe
Paris, October 2008
Scientific knowledge

Atlantic cod
(Gadus morhua)

Type of 3n Survival Animal Growth

shock (%) (%) stage performance
Thermal 87 10-20 Underway

-------

Meagre
(Argyrosomus regium)

Data not found

Scientific knowledge The future prospects for aquaculture breeding in Europe
Paris, October 2008

Turbot
(Scopthalmus maximus)
Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Cold 90-100 60-80 Juveniles 2n = 3n
Adults 3n > 2n
Inicial tests for commercial-scale : Spain

Halibut
(H.hipoglossus)
Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Cold 92-95 50-82 Juveniles 2n = 3n
Heat 84 20 Adults 2n > 3n
Inicial tests for commercial-scale : France

Sole
(Solea solea)

DATA NOT FOUND

Bottleneck: Quantity and Quality of the gametes
 The future prospects for aquaculture breeding in Europe
Scientific knowledge Paris, October 2008

Grass carp
(Ctenopharyngodon idella)
Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Cold 50-100 20 Juveniles 2n = 3n
Heat 87 50 2n > 3n
Pressure 100 70
Comercial-scale: USA

Nile tilapia
(Oreochromis niloticus) Type of 3n Survival Animal Growth
shock (%) (%) stage performance
Cold 83.3-100 variable Juveniles 2n = 3n
Heat - - 3n > 2n (separated)
Males > Females
Pressure - -
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 The future prospects for aquaculture breeding in Europe

Industrial application Paris, October 2008

Polyploidy in aquaculture

Species Country Application

Atlantic salmon Canada B, P
Rainbow trout Canada, France, Japan B, P
Sea bass (France) I
Sea bream - -
Atlantic cod - -
Meagre - -
Turbot (Spain) I
Halibut (France) I
Sole - -
Grass carp USA B, P*
Nile tilapia - -
Polyploidy applied in breeding programs (B), practical culture (P) or initial work (I).
*In the USA. In China, still experimental
 The future prospects for aquaculture breeding in Europe
Paris, October 2008

Practical knowledge gaps (I)

Cannulation

GAMETE MANIPULATION

9 Evaluation of broodstock

Assessment of normal development of the gametes before induction

9Spawning control (environmental/hormonal response of males and females)

Quality criteria for gametes after induction and before manipulation

9 Artificial fertilization approach

Male Female Artificial

stripping stripping fertilization

Graduated cylinder evaluation

 The future prospects for aquaculture breeding in Europe
Paris, October 2008
Practical knowledge gaps (II)

SHOCK TREATMENT

9 Evaluation of type of shock

Analysis of the three variables: Time-course-like experiments

Variability of results and feasibility of application

Scaling up from laboratory to commercial conditions

5 min DUR

Felip et al. (1997)

 The future prospects for aquaculture breeding in Europe
Paris, October 2008
Practical knowledge gaps (III)

TRIPLOIDY PROCEDURE

9 If an optimal shock treatment is achieved

It needs to be conducted each time in each generation

9 Evaluate the possibility to produce tetraploids

Males 4n produce sperm 2n (sperm size vs eggs micropile)

Females 1n x Males 2n = Offspring 3n

Females 4n produce eggs 2n (fertility)

Females 2n x Males 1n = Offspring 3n

Practical knowledge gaps (IV)
9 Evaluate triploid performance in terms of growth
TRIPLOIDY EFFECTS All-female stocks? Mixed-sex triploid stocks?

1200

1000 A D ip lo id s
Weight ( g )

800 T rip lo id s

600

400
P < 0.05
200

40
B
Length ( cm )

30

20 P > 0.05

10

1.5 C P < 0.05

Condition factor

1.4

1.3

1.2

1.1
0 5 10 15 20 25 30 35 40 45 50 55 60
A g e ( m o n th s )

Sea bass Turbot

Felip et al. (1999; 2001) Cal et al. (2006)
 The future prospects for aquaculture breeding in Europe
Paris, October 2008
10
Adults
8
N = 7
6 Performance of triploids is
species specific
(4)
4
(2)
2 (1)
0
Species Growth Reference
Number of cases

10 Juveniles
(8) Ictalurus punctatus Wolters et al. (1982)
8 (7) +
Oreochromis niloticus + Bramick et al. (1995)
N = 16
6 Heteropneustes fossilis Tiwary et al. (1997)
+
4 Oreochromis aureus + Valenti (1976)
Silurus glanis + Krasznai & Marian (1986)
2 (1) Oncorhynchus mykiss Solar et al. (1984)
-
0 Lepomis gibbosus - Kerby et al. (1995)
Oncorhynchus kisutch - Withler et al. (1995)
10 Monosex stocks
(All-females) Salmo salar - Galbreath et al. (1995)
8 Misgurnus mizolepis NC Kim et al. (1994)
Salmo salar NC McGeachy et al. (1995)
6 N = 3 Oreochromis niloticus NC Hussain et al. (1995)
4 (3) Oreochromis aureus NC Chang et al. (1993)
2 (+) positive, (-) negative, (NC) no change
Basant et al. (2004)
0
3n > 2n 3n = 2n 3n < 2n

Felip (2000)
TRIPLOIDY EFFECTS 9 Evaluate triploid performance in terms of reproduction

Felip et al. (2001)

Cal et al. (2006)

Degree and permanence of gonadal sterility in triploids

 The future prospects for aquaculture breeding in Europe
Future research axes Paris, October 2008

Evaluation of scapes for those species with interest in aquaculture

Development of protocols and scaling up to commercial conditions

Availability and costs of triploid production at a commercial-scale

(different farming facilities, biological performances, adverse
conditions, economical benefits)
 The future prospects for aquaculture breeding in Europe
Future research axes Paris, October 2008

Further research in tetraploidization and fertility

Little information about large older triploids (variation in market sizes)

Little information about field experiences with triploids (salmonids)

Further research in triploids as a model for basic research

(physiology, gene regulation, gametogenesis and infertility)

Public concern

Specific govermental regulation for the use of triploids in aquaculture

 The future prospects for aquaculture breeding in Europe
Paris, October 2008

STERILISING FISH TO PROTECT BIODIVERSITY AND IMPROVE QUALITY

Chromosome set manipulation for sterilising fish

Species specific problems & research gaps
Topic Sp1 Sp2 Sp3 Sp4
Control of induction/fertilization and
evaluation of gamete quality
Type of shock and analysis of variables
(optimum conditions and available devices)
Possibility of tetraploidy induction
Effects of triploidy: growth and reproductive
performance
Scaling-up to industry

Triploid verification (producers & users)

Other subjects