Does poly-ß-hydroxybutyrate stimulate the immune system

of European sea bass larvae?

Andrea Franke1, Olivia Roth1, Peter de Schryver2, Catriona Clemmesen1
1 GEOMAR
Helmholtz Center for Ocean Research, Kiel, Germany
afranke@geomar.de 2 Laboratory of Aquaculture & Artemia Reference Center, Ghent University, Belgium

BACKGROUND
 Mass mortality of fish larvae is a major bottleneck in aquaculture production
 Promising solution:
Application of dietary supplements such as prebiotics to improve the immunocompetence of larval fish

EXPERIMENTAL APPROACH
 Species in focus
 European sea bass (Dicentrarchus labrax) larvae were used from
28 days post hatch onwards
European sea bass larva ©fishbase.us

 Prebiotic in focus
 Poly-ß-hydroxybutyrate (PHB) is a bacterial energy storage compound
which was shown to have an immunostimulatory potential
(De Schryver et al., 2011, Environmental Microbiology 13(4), 1042-1051)

 Freeze-dried PHB-containing bacteria (Alcaligenes eutrophus) with a

© K. Thomas

Bacteria containing PHB inclusions

high PHB content (75%) were used

 Experimental timeline & design

 Sea bass larvae were reared in a flow-through system and fed with brine
28 Start experiment
Time (dph)

shrimp nauplii (Instar II) 3 times a day over a period of 10 days

31 1. Sampling
38 2. Sampling  Experimental groups (in triplicate):
1. PHB group: PHB-containing bacteria were encapsulated in brine shrimp
2. Control group: no PHB

GENE EXPRESSION: 2.5

Insulin-like growth factor (1. sampling)
2.0
Dicentracin = antimicrobial peptide (2. sampling)
F = 9.80, p < 0.05 F = 10.85, p < 0.05
RESULTS & CONCLUSION 2.0
x-fold gene expression relative to control

x-fold gene expression relative to control

1.5

1.5

1.0

1.0

0.5
0.5

0.0 0.0
Control PHB Control PHB

MHC class II alpha (2. sampling) MHC class II beta (2. sampling)
4.5 3.5
F = 14.35, p < 0.05 F = 8.27, p < 0.05
4.0
3.0
3.5
x-fold gene expression relative to control
x-fold gene expression relative to control

2.5
3.0

2.5 2.0

2.0 1.5

1.5
1.0
1.0
0.5
0.5

0.0 0.0
Control PHB Graphs display mean + SEM Control PHB