http://www.fao.org/documents/card/en/c/28b6bd62-5433-4fad-b5a1-8ac61eb671b1/ International Technical Seminar/Workshops on Acute hepatopancreatic necrosis disease (AHPND)

1. FAO TCP/INT/3502 “Reducing and managing the risk of
Acute Hepatopancreatic Necrosis Disease (AHPND) of Cultured Shrimp”
BREEDING FOR DISEASE
RESISTANCE IN SHRIMPS: A
tale of two pathogens
 Marcela Salazar, James Cock,
Morten Rye
 msalazar@ceniacua.org
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND: Government, Scientist and Farmer Responses”
22-24 June 2015, Tryp Hotel, Panama City 1

2. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 2

3. HOST DEFENSE
Resistance
Ability to limit
pathogen load.
Tolerance
Ability to “deal with”
a given pathogen
load.
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 3

4. A = Difference in resistance
B = Difference in tolerance
C= Both resistance and
Tolerance
D= No differences in
Resistance or tolerance
Science 2007. 318 pag 812
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 4

5. Decrease
risk of
cerebral
malaria
Increase
parasite
count
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 5

6. Resistance
Tolerance
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 6

7. EVOLUTIONARY
IMPLICATIONS Resistance = negative effect on pathogen prevalence
Tolerance = neutral or positive
Host resistance selects for parasite adaptation which then
selects for host resistance
= Evolutionary dynamics
A high rate of infection but low virulence should select for
host tolerance, whereas the opposite should favour
resistance (Restif & Koella 2004)
Tolerance is more likely to go to fixation than resistance (Roy
& Kirchner 2000). This is because resistance mechanisms
work by eliminating parasites, thus reducing the very
selection pressures that favoured them in the first place.
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 7

8. BREEDING FOR
DISEASE R/T
REAL BENEFIT OF
SELECTION
Genetic variation in
resistance
Standardized
methods for
selection of R/T
Correlation with
other traits
Genotype x
Genotype
interaction
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 8

9. MEASURING RESISTANCE /
TOLERANCE
Experimental challenge
 Infection route
 Minced tissue
 Per os
 Reverse gavage
 Injection
 Immersion
 Conditions
 Temperature
 Density
 Oxygen
 Statistical analysis
 Binary trait
 Survival curve analysis
 Hazard model
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 9

10. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 10

11. Sobrevivencia prueba de desafio
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0
0
10
20
30
40
50
60
70
80
90
100
Canastas
Libres
days
Percentsurvival
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 11

12. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 12

13. TAURA SYNDROME VIRUS
Hit Colombia 1994
Pond survivors as
breeders
increased survival
by 30%
Family selection
started in 1997
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 13

14. TYPICAL CHALLENGE TEST
% Mortalidad diaria
0,00%
2,00%
4,00%
6,00%
8,00%
10,00%
12,00%
14,00%
1
3
5
7
9
11
13
15
17
19
21
23
25
Dias Post-inf
%
Tanque 1
Tanque 2
Tanque 3
0,00%
10,00%
20,00%
30,00%
40,00%
50,00%
60,00%
70,00%
80,00%
90,00%
100,00%
81 12 23 28 18 86 80 69 2 56 46 87 18 54 9 85 73 8 7 71 70 48 58
Flias
% Sopervivencia Familias L20
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 14

15. TSV vs. pond survival
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 15
PC 1-2 PC 11-14 SI R6 Farm 1 Farm2
PC 1-2 0.83** 0.70** 0.50* 0.51*
PC 11-14 0.20 0.72** 0.42* 0.54*
SI R6 0.15 0.28 0.42* 0.38*
Farm 1 0.17 -0.10 0.03 0.50**
Farm 2 -0.07 0.22 -0.03
TSV CT 0.12 0.16 0.12 0.63** 0.03

16. OCEANIC INSTITUTE TSV
1995-1997
 One line combining two
selection traits: Growth and
TSV
 Phenotypic and genetic
negative correlation between
the 2 traits
1998
 Two separate lines
 Growth 100%
 21% increase in one
generation
 TSV (70% TSV – 30%
growth)
 Gain in 18.4% survival per
generation
 h2 = 0.30
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 16

17. # molts (26 days) Molt cycle
TSV susceptible 3.25 ± 0.72 7.49 ± 1.71
TSV resistant 2.60 ± 1.10 9.64 ± 3.41
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 17

18. If resistance is negatively
correlated with growth and
reproductive capacity it will be
rapidly lost from the breeding
nucleus unless continuous
selection pressure is maintained
for resistance.
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 18

19. 20
40
50
60
70
80
90
30
1993
1994
1997
1999
2004
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 19

20. 
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 20

21. GENOTYPE X GENOTYPE
INTERACTION
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 21

22. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 22

23. “ A piece of bad news
wrapped up in a protein
coat”
Hit Colombia
1999
Family
selection
2000-2004
Mass
selection
from 2008
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 23

24. 0
0,5
1
0 5 10 15 20 25 30
SDF
Batch 8 full dose
Batch 9 full dose
Batch 10 diluted dose
Batch 11 diluted dose
Batch 13 diluted dose*
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 24

25. SUMMARY OF RESULTS
BATCHES 1-25
TRAIT h2 c2
Harvest weight 0.44 ± 0.02 0.07 ± 0.05
Pond survival 0.07 ± 0.01 0.03 ± 0.01
WSSV resistance 0.03 ± 0.01 0.02 ± 0.01
TSV resistance 0.19 ± 0.09 0.07 ± 0.01
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 25

26. International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City
GENETIC CORRELATIONS
Familias
-0.01
-0.005
0
0.005
0.01
0.015
0.02
-1.5 -1 -0.5 0 0.5 1 1.5
BV Crecimiento
BVparaResistenciaaWSSV
Lote Corr
Gen
4 -0.23
6 -0.31
3 -0.54
5 -0.35
7 -0.47
8 -0.25
24/06/2015
26

27. CHANGE OF STRATEGY
Millions of animals infected
Several different origins
Including wild populations
Infection at PL 40 stage
First challenge survival 1x10-5
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 27

28. 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
0
10
20
30
40
50
60
70
80
90
100
Dias
Percentsurvival
TUMACOCARTAGENA
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 28

29. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 29

30. ADAPTIVE IMMUNITY
INVERTEBRATES
 Viral accommodation TW Flegel
 Several studies showing immune priming in
invertebrates
 Some reports of transgenerational priming in insects
 Maternal
 Paternal
 DSCAM molecules in shrimp associated with specific
response to certain bacteria.
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 30

31. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 31

32. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 32

33. Growth
Reproduction
Immunity
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 33

34.  Negative genetic correlation between tolerance and
fitness in the absence of parasites.
 Identified in plants, Insects, Shrimps, Vertebrates
 Allocation of resources
 Autoimmunity
 Inbred strains longevity and Sendai infection
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 34

35. INBREEDING
BREEDERS – full diversity
high quality, low inbreeding
HATCHERIES
copy
HATCHERIES
subsample
FARMS
restricted subsample
subsample
zero inbreeding inbred epidemics
Inbreeding, disease and epidemics Roger Doyle 2014
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 35

36. INBREEDING
 Inbred strains of mice a
valuable tool for research
 Inbreeding can help in the
fixation of very low frequency
genes such as WSSV
resistance
 Genes can be introgressed
 Successful crops highly
inbred
 Genetic variability in
resistance key to breeding
selection
 Mating selection based in
differences in MHC
 Mega variability in MHC
alleles (>10,000 alleles)
explained by
 Heterocigote advantage
 Overdominant selection
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 36

37. NEGATIVE CORRELATION?
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 37

38. CONCLUSIONS
 Control measures
 Environment
 Microbiota
 Other ???
 Determine genetic variability in survival against the
disease in several different populations
 Develop appropriate challenge tests
 Toxin
 V parahaemolyticus
 Make use of genomic tools to find markers
 Mass selection – IMMUNE PRIMING. Clean breeders
24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 38

39. 24/06/2015
International Technical Seminar/Workshop “EMS/AHPND:
Government, Scientist and Farmer Responses” 22-24 June 2015,
Tryp Hotel, Panama City 39