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Batam Mariculture Development Center, Directorate General of Aquaculture, Ministry of Marine Affairs
and Fisheries, Republic of Indonesia
2)
Master Student at Human Nutrion and Rural Development, Faculty of Bioscience Engineering,
University of Ghent
3)
Aquativ (DIANA Division, Member of SYMRISE Group), ZA du Gohlis 56250 Elven, France
*: Corresponding author : Romi Novriadi, Jl. Raya Barelang Jembatan III, Pulau Setoko, Batam 29422.
E-mail: Romi_bbl@yahoo.co.id
ABSTRACT
We investigated the effects of fish protein hydrolysate (FPH) on zootechnical performance and
immune response of the Asian Seabass Lates calcarifer Bloch. Experimental fish were fed with
3 diets: a local commercial diet (control), coated or not, with 2 and 3% FPH (w/w). Twelve
thousand Asian Seabass juveniles (5.880.56 g) were divided into three groups and two
replicates reared in nursery tanks (2000 L). The remaining fish were then used for grow-out
experiment in floating net cages (1m x 1 m x 3 m). Zootechnical performances were assessed
at both stages with following indicators: total weight gain (TWG), % relative weight gain (%
RWG), % specific growth rate (% SGR), final weight (g) and final length (cm). At the end of each
trial period, fish immune status was assessed through blood sampling and the measurement of
Neutrophile (%), Monocyte (%), Lymphocyte (%), Macrophage (105 cell/mL), Leukocyte (103
cell/mL) and Phagocytes activity (%). At the end of the nursery trial, an immersion bacterial
challenge with Vibrio parahaemolyticus (105 cells mL-1) was implemented. The results showed
that dietary FPH supplementation significantly influenced the growth and immune status of
Asian Seabass when compared to the control group. Fish fed FPH supplemented diet yielded
higher growth rates and survival rates than non supplemented group. Fish phagocytic activity
and resistance to a bacterial challenge were also improved by dietary FPH supplementation.
These results may be related to the significant changes observed in fish leukocyte profiles,
when fed FPH supplemented diets. Altogether, these results show the positive contribution of
FPH to the sustainability of Asian seabass farming.
Keywords: Functional hydrolysates, Asian Seabass, Zootechnical performance, Immune
response, Phagocytic activity, Nursery, Grow out
INTRODUCTION
In Asian Seabass culture, disease
outbreaks are being increasingly reported as
a major constraint to the sustainable growth
of production (Chong et al., 1987; Bloch and
Larsen, 1993; Chou et al., 1998). Many
diseases are linked to the stress conditions
associated with the intensification systems
of farming and the degradation of the
environmental
quality.
Under
poor
conditions, live food and larvi are often
opportunistically infected by fungi, bacteria
and viruses. Conventional treatment of
aquatic diseases through the use of
disinfectants and antibiotics to overcome the
bacterial infection problem are having limited
value and has stimulated the development
of bacterial resistance (Defoirdt et al., 2007;
Subasinghe, 1997; Cabello, 2006). Another
Design
and
Diet
Growth Performance
In the current study, the influence of
functional hydrolysates in Asian seabass
Table 1. Growth performances of L. calcarifer fed dietary treatments for 6 weeks during the
nursery phase.
Treatments
TWG (g)
AQUATIV 3
RWG (%)
SGR (%)
18.250.47a
530.3544.39a
6.570.25a
Final weight
(g)
21.710.39a
Final length
(cm)
11.760.21a
16.640.45b
483.6539.81b
6.290.25a
20.100.39a
11.000.17a
9.870.89c
287.1335.99c
4.820.34b
13.330.84b
10.330.32b
% FPH
AQUATIV 2
% FPH
Control
TWG: Total Weight Gain (g), RWG: Relative Weight Gain (%), SGR: Specific Growth Rate
Significant differences among the treatments and control are indicated by different letter (n=2, P<0.05).
Following from this initial nursery phase, the dietary test went on for another 8 weeks in grow-out units.
Table 2. Growth performances of L. calcarifer fed dietary treatments for 9 weeks during the
grow-out phase.
Treatments
TWG (g)
RWG (%)
SGR (%)
AQUATIV 3
40.741.23a
187.807.40a
1.920.05a
62.941.47a
Final length
(cm)
16.560.23a
36.762.60b
182.9913.84a
1.890.09a
56.942.26b
15.550.28b
26.311.99c
198.7726.21a
1.980.15b
39.601.49c
13.770.31c
% FPH
AQUATIV 2
% FPH
Control
TWG: Total Weight Gain (g), RWG: Relative Weight Gain (%), SGR: Specific Growth Rate
Significant differences among the treatments and control are indicated by different letter (n=2, P<0.05).
During the grow-out feeding trial, SGR were lower for supplemented diet fish groups due to their higher
average weight and length. TWG however remained higher for fish groups receiving FPH supplemented
diets.
Immune Performance
Blood white cell counts are detailed in Table 3 (nursery) and Table 4 (grow out).
Table 3. Cellular immune response of L. calcarifer fed dietary treatments for 6 weeks during
the nursery phase.
Parameter
FPH Treatments
Control
2%
b
3%
Neutrophile (%)
5.070.19
6.310.15
6.730.12a
Monocyte (%)
1.930.2b
2.650.1a
2.80.13a
50.871.49b
63.741.19a
66.710.71a
Lymphocyte (%)
5
1.830.25
2.620.12
2.830.15a
48.331.36b
55.671.14a
56.341.17a
15.771.18b
37.951.01a
40.230.52a
Significant differences among the treatments and control are indicated by different letter (n=6, P<0.05).
Table 4. Cellular immune response of L. calcarifer fed dietary treatments for 9 weeks during
the grow-out phase.
Parameter
FPH Treatments
Control
2%
3%
Neutrophile (%)
6.030.12
6.600.10
6.630.12
Monocyte (%)
2.400.10
2.870.12
2.930.06
55.210.76
65.251.09
66.301.07
2.300.20
2.600.10
2.670.12
49.680.56
57.012.36
54.670.43
28.290.67
39.641.03
40.480.95
Lymphocyte (%)
5
Significant differences among the treatments and control are indicated by different letter (n=6, P<0.05).
Survival Rates
As illustrated by Figure 1, at the end
of nursery feeding trial, the 2 and 3%
hydrolysate supplementations resulted in
96.750.28 % and 97.280.18 % survival
rate respectively, while the control diet
yielded 93.650.13 %. More contrasted
Figure 1. Histogram of the mean percentage survival (%) of L. calcarifer during experiment
period in nursery phase (a) and grow out phase (b). Significant differences among the
treatments and control are indicated by different letter (p< 0.05).
Figure 2. Histogram of the mean survival (%) of L. calcarifer (n=20, weight: 20g) challenged
with V. parahaemolyticus at 105 cells/ml. Survival was scored after 5 days challenge with V.
parahaemolyticus. Significant differences among the treatments and control are indicated by
different letter (n=2, p< 0.05).
Water Quality Analysis
Table 5 Water quality analysis during AQUATIV nursery feeding trial. Sampling campaign was
performed on weekly basis at 9 AM at the middle point of experimental cages.
Paramet
er
Total
Bacteria
Total
Vibrio
CFU/m
L
CFU/m
L
pH
Nitrate
(NO3)
Nitrite
(NO2)
Ammonia
(NH3)
Posphate
(PO4)
Salinity
Test Results
Unit
2%
1.1 2.7x102
1.4 2.7x102
0.2 1.1x102
1.4 2.7x102
7.76 8.25
7.72 8.18
7.73 8.19
<0.01
<0.01
<0.1
<0.1
<0.02 0.094
<0.02 0.094
0.018-0.027
0.015-0.022
30 31
30 - 31
mg/L
<0.01
mg/L
<0.1
mg/L
<0.02 0.094
mg/L
0.0120.024
Method
Specification
Control
30 31
Turbidity
NTU
0.65 1.42
Temperat
ure
30.1 30.4
3%
1.1
2.8x102
1.1
2.8x102
0.65 1.42
0.81 1.44
30.1 30.4
30.1 30.4
IKM/5.4.10/BBLB
Convensional
SNI 06-6989.112004
Colorimetry
Colorimetry
IKM/5.4.6/BBL-B
IKM/5.4.8/BBL-B
IKM/5.4.4/BBL-B
IKM/5.4.9/BBL-B
Thermometer
Table 6. Water quality analysis during AQUATIV grow-out feeding trial. Sampling campaign
was performed on weekly basis at 9 AM at the middle point of experimental cages.
Test Results
Parameter
Unit
15 Juli
Total
Bacteria
Total
Vibrio
CFU/
mL
CFU/
mL
22 Jul
29 Jul
5 Aug
12 Aug
19
Aug
26 Aug
173
1.2x103
5.2x103
2x102
23
131
2.7x102
8.18
7.78
8.26
1.4x1
03
3.4x1
02
7.76
3.7x103
8.18
2.7x1
02
3.1x1
02
8.37
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.1
<0.1
<0.1
<0.1
<0.1
<0.1
<0.1
<0.009
<0.009
<0.009
0.357
<0.009
<0.009
0.027
0.025
0.047
<0.033
30
30
31
<0.03
3
30
30
<0.00
9
<0.03
3
31
7.15
2.63
5.33
27
2.44
6.52
7.26
29.1
30.5
28.7
29.6
30.5
29.7
30.7
1.9x102
1.9x102
8.25
pH
Nitrate
(NO3)
Nitrite
(NO2)
Ammonia
(NH3)
Posphate
(PO4)
Salinity
Turbidity
Temperatu
-re
mg/L
mg/L
mg/L
mg/L
NTU
C
<0.033
31
Metoda
Analisa
IKM/5.4.10
/BBL-B
Convensio
nal
SNI
066989.112004
Colorimetr
y
Colorimetr
y
IKM/5.4.6/
BBL-B
IKM/5.4.8/
BBL-B
IKM/5.4.4/
BBL-B
IKM/5.4.9/
BBL-B
Thermom
eter
CONCLUSION
In conclusion, this work revealed that
the addition of functional hydrolysates in
Asian Seabass feed was able to enhance
significantly the fish immune system as well
as the fish zootechnical performances such
as Survival rate, total weight gain (TWG),
relative weight gain (RWG) and specific
growth rate (SGR) during the experimental
period. Therefore, dietary supplementation
of functional hydrolysates in Asian Seabass,
Lates calcarifer feeds should be encouraged
as a holistic approach to improving its
sustainable culture.
ACKNOWLEDGMENTS
This study was supported by SPF
DIANA (Member of SYMRISE Group),
through its AQUATIV Division by providing
operational fund, commercial feed and
functional hydrolysates to conduct field test
in Batam Mariculture Development Center.
We are thankful to the Reference laboratory
of Fish Diseases and Environmental
Analysis Serang for their assistance in
carrying out the histology analysis and
BMDC Laboratory staff for their assistance
in carrying out the water quality and
bacteriology analysis.
REFERENCES
Adibi,
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