Aquaculture Nutrition
doi: 10.1111/j.1365-2095.2010.00807.x
2011 17; e615–e621
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1,2
2
2
3
1
1
2
2
1
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China; 2 Nutrition
Laboratory, Institute of Aquatic Economic Animals, School of Life Science, Sun Yat-sen University, Guangzhou, China;
3
Laboratory of Aquaculture Nutrition, College of Fisheries, Ocean University of China, Qingdao, China
One experiment was conducted to determine the nutritive
value of phospholipids on growth performance of early
postlarval shrimp, Litopenaeus vannamei. Five isoenergic and
isonitrogenous diets with five supplemented levels of phospholipids (P1, P2, P3, P4 and P5 with 0, 10, 20, 40 and
80 g kg)1 diet, respectively) were fed to triplicate groups of
L. vannamei (mean initial wet weight 0.8 mg) for 27 days.
After the 27-day feeding trial, the lowest weight gain (WG,
%) and specific growth rate (SGR, % day)1) was found in P1
treatment, the highest WG and SGR was found in P3, P4 and
P5 treatments, P2 treatment provided intermediate result and
showed significant difference compared to P1, P3, P4 and P5
treatments. Shrimp fed the P1 diet had significantly lower
survival than shrimp fed other diets, while no significant
difference was found in survival among P2, P3, P4 and P5
treatments. Broken-line analysis on WG indicated that the
optimum dietary phospholipids for early postlarval shrimp,
L. vannamei, is 45.96 g kg)1 diet.
KEY WORDS: growth performance, Litopenaeus vannamei
larvae, phospholipids requirement
Received 7 February 2010, accepted 20 May 2010
Correspondence: Li-Xia Tian, Nutrition Laboratory, Institute of Aquatic
Economic Animals, School of Life Science, Sun Yat-sen University,
Guangzhou 510275, China. E-mail: [email protected]
Phospholipids are major constituents of membranes and are
vital to the normal function of every cell and organ. They
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2010 Blackwell Publishing Ltd
maintain cell structure and function and have regulatory
activities within the membrane and outside the cell. Without
phospholipids in membranes, there is neither cell respiration
occurring in the mitochondria nor mobility of the membranes (Mu 2005). For instance, phospholipids serve as second messengers in cell signalling, which is an essential
process in regulating cell growth, proliferation, differentiation, metabolism, nutrient uptake, ion transport, and even
programmed cell death (DÕAbramo et al. 1981). In addition,
there is evidence that phospholipids containing choline,
sphingomyelin, and their metabolites are important mediators and modulators of transmembrane signalling (Zeisel
1993). Phospholipids act as emulsifiers and facilitate the
digestion and absorption of fatty acids, bile salts and other
lipid-soluble matters. They also have a role in the transport
of lipids. Phospholipids act not only in the transport of absorbed lipids from the gut epithelium into the haemolymph,
but also in the transport of lipids between tissues and organs
(Coutteau et al. 1997) because they are constituents of lipoproteins. Even though some crustaceans can synthesize
phospholipids (Shieh 1969), their biosynthesis generally
cannot meet their metabolic demand (Kanazawa et al. 1985).
The importance of phospholipids in penaeid shrimp nutrition, including Litopenaeus vannamei (Boone), has been
demonstrated by many researchers (Coutteau et al. 1996,
2000; Paibulkichakul et al. 1998; Gong et al. 2000a, 2001;
González-Félix et al. 2002a).
Beneficial effects on culture performance as a result of
the inclusion of phospholipids in diets were first reported in
the late 1970s for lobster (Conklin et al. 1977) and prawn
(Kanazawa et al. 1979). While most studies of dietary lecithin
requirements have focused on juvenile and larger shrimp
rather than larval and early postlarval shrimp. Coutteau et al.
(1996) reported that the growth response of L. vannamei
postlarvae was significantly improved by the addition of 1.5%
soybean phosphatidylcholine (PC, 95% purity) or 6.5% deoiled soybean lecithin (23% PC) to the diet. Paibulkichakul
et al. (1998) indicated that 1.0% lecithin may be suitable for
P. monodon postlarval requirement. Coutteau et al. (2000)
indicated that the addition of any PC types (1.5% SPC from
soybean lecithin or 1.5% MPC from marine fish roe) resulted
in a significant increase in growth and decrease in sensitivity
to osmotic stress compared to PC-deprived shrimp. Gong
et al. (2000a) observed that juvenile L. vannamei growth was
enhanced as the level of phospholipids increased from 0 to
50 g kg)1 diet. However, as the level of dietary cholesterol
increased from 0 to 4 g kg)1 diet, the growth-promoting effect
of phospholipids diminished. Gong et al. (2001) evaluated
different types of soybean lecithin and their dietary requirement for juvenile L. vannamei in two 6-week growth trials.
Under their experimental conditions, there were no significant
interactions detected between lecithin type and phospholipid
level on shrimp growth or survival. However, shrimp growth
increased with phospholipids levels up to 3–5% of diet.
Hence, for juvenile L. vannamei, they recommended a supplementation level of phospholipids from 3% to 5% of diet.
González-Félix et al. (2002a,b) evaluated the effect of dietary
phospholipids on essential fatty acid (EFA) requirements of
juvenile L. vannamei and their potential interaction in a
6-week trial. A 3 · 3 factorial experiment was carried out
with increasing levels of soybean lecithin as the dietary
phospholipids (0%, 1.5%, or 3% of diet) and three dietary
levels (0%, 0.25%, or 0.5% of diet) of docosahexaenoic acid
(DHA) or n-3 highly unsaturated fatty acid (HUFA). No
significant interactions between the effects of phospholipids
and DHA or n-3 HUFA on growth were detected under
their experimental conditions; however, higher dietary inclusion level (0.5% of diet) of DHA did not further improve
growth and appeared to have a detrimental effect on survival
of shrimp. Meanwhile, high level (3% of diet) of dietary
phospholipids significantly improved growth of shrimp. The
optimal levels of phospholipids are dependent upon the
species, age and phospholipids fraction. In general, the
phospholipids requirement decreases with age or developmental stage of shrimp, with the larval stage very sensitive to
dietary phospholipids deficiency (Mu 2005).
The phospholipids requirement of postlarval L. vannamei
is not yet established. Moreover, a wide variety of phospholipids sources and phospholipids purity have been
employed in the previously cited nutritional studies. This
makes it difficult to draw conclusion with regard to the
optimal levels/types of dietary phospholipids required and to
compare studies performed by different investigators using
different shrimp species. Therefore, the objectives of this
study were to evaluate the nutritive values of phospholipids
for early postlarval L. vannamei.
Five artificial diets (P1, P2, P3, P4 and P5) were prepared by
supplementing phospholipids at 0, 10, 20, 40 and 80 g kg)1 in
each diet, as shown in Table 1, respectively. The phospholipids (provided by Jiakangyuan Beijing Company Ltd,
Beijing, China) are a product refined from soybean lecithin.
Chemical analyses indicated that phospholipids purity is
97% and contains 60% PC.
Table 1 Ingredients and proximate composition of the five experimental diets (g kg)1 dry matter)
Ingredients
White fish meal1
Protein hydrolysate2
a-Starch
Soybean oil3
Phospholipids (purity
97%, pc-60)4
Vitamin premix5
Mineral premix6
Vitamin C
Krill meal
Beer yeast
Cholesterol (purity 95%)
Others7
Composition
Phospholipids
Moisture
Crude protein
Crude lipid
Ash
1
2
3
4
5
6
7
P1
P2
P3
P4
P5
487.5
200
50
80
0
487.5
200
50
70
10
487.5
200
50
60
20
487.5
200
50
40
40
487.5
200
50
0
80
10
40
6.5
30
30
10
56
10
40
6.5
30
30
10
56
10
40
6.5
30
30
10
56
10
40
6.5
30
30
10
56
10
40
6.5
30
30
10
56
27.2
63.5
567.2
173.1
169.5
36.5
73.8
570.0
171.3
169.1
45.8
79.0
565.8
170.7
168.4
64.4
64.2
564.4
172.5
170.7
101.6
69.4
562.5
170.8
170.4
Imported from Australia.
Huaqi Guangzhou Company Ltd, Guangzhou, China.
Guangzhou Donghao Company Ltd, Guangzhou, China.
Jiakangyuan Beijing Company Ltd, Beijing, China.
Contained (g kg)1) retinyl acetate, 2.5 g; cholecalciferol, 6.25 g;
all-rac-a-tocopheryl acetate, 75 g; menadione, 2.5 g; thiamin,
0.25 g; riboflavin, 1 g; D-calcium pantothenate, 5 g; pyridoxine
HCL, 0.75 g; cyanocobalamin, 2.5 g; niacin, 2.5 g; folic acid,
0.25 g; biotine, 2.5 g; meso-inositol, 379 g; cellulose, 500 g (Niu
et al. 2008).
Contained (g kg)1) KCl, 90 g; KI, 40 mg; NaCl, 40 g;CuSO4-5H2O,
3 g; ZnSO4-7H2O, 4 g; CoSO4-7H2O, 20 mg; FeSO4-7H2O, 20 g;
MnSO4-H2O, 3 g; MgSO4-7H2O, 124 g; Ca(HPO4)2-2H2O, 500 g;
CaCO3, 215 g. (Niu et al. 2008).
Contained (g kg)1): sodium alginate, 30; lysine, 1 g; choline
chloride, 10 g; methionine, 10; tryptophan, 5.
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Aquaculture Nutrition 17; e615–e621 2010 Blackwell Publishing Ltd
The method of diet preparation was the same as in Niu
et al. (2009). Briefly, all the dry ingredients for each of the
experimental diets were weighed, combined and thoroughly
mixed to a homogenous consistency in a Hobart-type mixer.
Then, the soybean oil was added and thoroughly mixed for
5 min. Deionized water (250 mL kg)1 dry ingredients mixture) was added and mixed for another 5 min. The wet
mixture was placed in a monoscrew extruder (Institute of
Chemical Engineering, South China University of Technology, Guangzhou, China) and extruded through a 1.2-mm die.
The resulting pellets were dried at 25 C with the aid of an air
conditioner and an electrical fan. After drying, the diets were
broken up and ground using a mortar and pestle and graded
through a series of different-sized metal sieves. Shrimps were
acclimated to the experimental conditions and fed the control
diet (without supplemented phospholipids) at a particle size
of 300 lm for 3 days before the start of the experiment. The
particle size changed to 450 lm, 600 lm, 900 lm and
1.2 mm, respectively, from days 1 to 5, 6 to 10, 11 to 21 and
22 to 30. All the diets were stored at )20 C until used.
A 27-day feeding trial was conducted in a recirculating water
system. The system consisted of 15 aquaria (150 ·
70 · 60 cm) supplied with 500 L continuous filtered seawater
circulating constantly. Each aquarium was connected with a
separate recirculating system. Each recirculating system was
equipped with a valve and could modulate the flow rate of
water depending on the shrimp developmental condition.
The seawater was continuously recirculated by two air–water
lifts at a rate of 20 L min )1 (when the valve was opened
completely). Each recirculating system was equipped with a
sand filter and packed-column biological filter. The water
was changed from the separate recirculating system according to the water quality, and about one-third (150–200 L)
new water was added into the recirculation system daily.
During the trial, the diurnal cycle was 15 h light/9 h dark.
Water quality parameters were recorded daily and were
maintained as follows: salinity, 30–32 g L)1; temperature,
27–29 C; dissolved oxygen, 5.6–6.2 mg L)1; ammonia–
nitrogen, 0.05–0.07 mg L)1.
The shrimp used in this experiment were obtained from
Evergreen (Zhanjiang) South Ocean Science and Tech Co.
Ltd, Zhanjiang, China, and the early postlarvae were
obtained just after they metamorphosized from mysid stage.
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Aquaculture Nutrition 17; e615–e621 2010 Blackwell Publishing Ltd
Shrimps were collected randomly, and groups of 100 shrimps
were weighed (shrimps were fasted for 24 h and then weighed)
before being stocked into individual tanks. Initial average wet
weight (0.8 mg) was calculated by dividing the group weight
by the number of shrimps. Three replicate tanks (with 1000
shrimps initially in each tank) were used for each dietary
treatment. Shrimps were fed the experimental diets six times
daily (07:00, 10:00, 13:00, 16:00, 19:00 and 22:00 h). Feeding
quantity was adjusted so that shrimps were fed to slight
excess. Uneaten feed, faecal waste and moult exuviate were
removed automatically by the separate recirculating system
after the nightly circulation. At the termination of the feeding
trial, all surviving shrimps from each tank were also weighed
as a group. Final weights were calculated by dividing the
group weight by the number of shrimp. Survival was calculated by individually counting of all the surviving shrimp at
the beginning and at the end of the experiment.
At the end of feeding trial, shrimps were fasted for 24 h and
then weighed. After weighting, all shrimps in each tank were
used for samples, dried and ground for whole body composition and lipid analysis. Lipid were extracted with chloroform–methanol (Folch et al. 1957) from the whole body of
shrimp and then further separated into neutral lipid and
polar lipid fractions by Sep-pak silica cartridge (Juaneda &
Rocquelin 1985). Both fractions were analysed for lipid
classes by an Iatroscan in Sun Yat-Sen University of Madical
Sciences. Lipid classes were identified in comparison with
standard (Sigma, St. Louis, MO, USA). Moisture, crude
protein and ash of the experimental diets and shrimps were
determined using standard methods (AOAC 1984). Moisture
was determined by drying in an oven at 105 C for 24 h;
crude protein was analysed by Kjeldahl method after acid
digestion (1030-auto-analyzer; Tecator, Höganäs, Sweden).
Oven-dried feed and whole body were ashed at 550 C for
24 h in a muffle furnace.
Optimal dietary phospholipids were determined using weight
gain by the broken-line model (Robbins et al. 1979). Other
data from triplicate tanks of each diet were analysed using
one-way analysis of variance and DuncanÕs multiple-range
test. The software was SPSS (Version 10.0, SPSS Inc., Chicago, IL, USA). Differences were considered significant at
P < 0.05.
Weight gain (WG), specific growth rate and survival are
summarized in Table 2. Survival of postlarvae in P1 treatment was significantly lower than that of shrimp in P2, P3, P4
and P5 treatments and was in the range of 63–89%. Growth
performance of shrimp in P3, P4 and P5 treatments were
significantly higher than that of shrimp in P1 and P2 treatments (P < 0.05). No significant difference was found in
growth performance of shrimp among P3, P4 and P5 treatments. Increased weight gain is one of the beneficial effects of
supplementing phospholipids to the diet of shrimp and has
been well documented for many species, such as P. monodon
(Piedad-Pascual 1986), P. vannamei (Coutteau et al. 1996),
P. monodon (Paibulkichakul et al. 1998) and P. vannamei
(Coutteau et al. 2000). However, significant effect of phospholipids supplementation on survival has only been demonstrated in some other larval and postlarval stages of
shrimp, such as M. japonicus (Kanazawa et al. 1985) and
P. monodon (Paibulkichakul et al. 1998). Juvenile shrimp
appear to be less sensitive to deficiency of phospholipids,
indicating that the phospholipids requirement diminishes in
juvenile stages, possibly because of a more developed digestive tract and enhanced activity of lipolytic enzymes or an
increased capacity for de novo synthesis of phospholipids in
L. vannamei (Coutteau et al. 1996; Gong et al. 2001; González-Félix et al. 2002a) and P. merguiensis (Thongrod &
Boonyaratpalin 1998). In this experiment, dietary supplemented phospholipids significantly improved the growth
performance of early postlarval L. vannamei. It could indicate that early life stages of shrimp are not capable of
synthesizing phospholipids at a rate sufficient to meet the
requirement for formation of new cell components during the
initially short period of rapid growth. Moreover, survival was
also improved by supplementing phospholipids in the present
experiment. Geurden et al. (1998) declared that the phospholipids composition of PC has an early growth-promoting
effect preceding developmental abnormalities with phosphatidylinositol (PI) almost completely preventing the deformities. PC and PI are more effective in promoting growth and
survival of larval and juvenile shrimp and are regarded as
active phospholipid fractions. The significantly better initial
growth and survival in the phospholipids-supplemented
treatments may be because of the components of PC and PI.
A dietary phospholipids could be superior to supplementation with a single purified class in aquatic diets (Geurden
et al. 1998). According to Coutteau et al. (1997), dietary
phospholipids may serve as a source of choline, inositol, EFA
and energy. Other studies have indicated that dietary phospholipids may improve the efficiency of EFA supplied as
neutral lipid. This would be because of a more efficient
transport and better lipid mobilization from the hepatopancreas to the haemolymph and to other tissues and organs,
resulting in enhanced lipid deposition and increased energy
availability for growth (Teshima et al. 1986a,b; Kontara
et al. 1998).
Optimal inclusion levels of dietary phospholipids reported
for various species of juvenile penaeids range from 12.5 to
30 g kg)1 diet, such as juvenile P. penicillatus: 12.5 g kg)1
diet (Chen & Jenn 1991), juvenile P. monodon: 12.5 g kg)1
diet (Chen 1993), juvenile P. chinensis: 20 g kg)1 diet
(Kanazawa 1993) and juvenile P. japonicus: 30 g kg)1 diet
Table 2 Growth performance of shrimp fed diets with and without supplementation of phospholipids
One-way
Dietary phospholipids
levels (g kg)1)
P1
27.2
P2
36.5
P3
45.8
P4
64.4
P5
101.6
(P-value)
Growth performance
Initial number
IBW (mg)
Final number
FBW (mg)
WG (%)
SGR (% day)1)
Survival (%)
1000
0.8
625.7 ±
38.4 ±
4705 ±
14.3 ±
62.6 ±
1000
0.8
875.7 ±
41.4 ±
5075 ±
14.6 ±
87.6 ±
1000
0.8
891.0 ±
43.8 ±
5370 ±
14.8 ±
89.1 ±
1000
0.8
877.3 ±
44.0 ±
5406 ±
14.8 ±
87.7 ±
1000
0.8
885.3 ±
44.3 ±
5441 ±
14.9 ±
88.5 ±
–
–
0.000
0.001
0.001
0.001
0.000
60.0b
1.0c
124c
0.1c
6.0b
8.4a
0.4b
44b
0.1b
0.8a
13.7a
0.9a
110a
0.1a
1.4a
9.1a
0.5a
64a
0.1a
0.9a
ANOVA
19.9a
0.5a
59a
0.2a
2.0a
Values are shown as means ± SE of three replicates. Values within the same row and not sharing a common superscript are significantly
different (Ducans, P < 0.05).
IBW (mg shrimp)1): initial body wet weight (mg shrimp)1).
FBW (mg shrimp)1): final body wet weight (mg shrimp)1).
WG (%): weight gain = 100 · (final body weight – initial body weight)/initial body weight.
SGR (% day)1): specific growth rate = 100 · (ln final wt. – ln initial wt.)/total number of experimental days.
Survival (%) = 100 · (final shrimp number)/(initial shrimp number).
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Aquaculture Nutrition 17; e615–e621 2010 Blackwell Publishing Ltd
6000
y = 35.784x + 3743.6
R = 0.91
WG (%)
5500
y = 0.9511x + 5344.6
R = 0.18
5000
4500
Xopt = 45.96
Figure 1 Optimum phospholipids level
based on weight gain as determined by
the broken-line model.
4000
0
20
40
60
80
100
120
–1
Dietary phospholipids (g kg diet)
(Teshima et al. 1986a). While for juvenile L. vannamei, Gong
et al. (2001) recommended a supplementation level of phospholipids from 30 to 50 g kg)1 diet. In our experiment,
broken-line analysis on weight gain indicated that the optimum phospholipids requirement of early L. vannamei postlarvae is 45.96 g kg)1 diet (Fig. 1). The phospholipids
requirement for postlarval L. vannamei in this experiment
using the common broken-line regression analysis is a little
higher than that for the above-mentioned shrimp juveniles. A
comparison between different sources of phospholipids
within a study for an individual aquatic species should be
limited to comparing the potency of each source of phospholipids from different studies because the experimental
conditions such as the duration of the study, the developmental stage of shrimp and the water temperature often
differ. Moreover, the larval/postlarval shrimp have higher
nutritional requirements than juvenile shrimp. These may be
the reasons that the phospholipids requirement for early
postlarval L. vannamei in this experiment is a bit higher than
that of other published data.
Table 3 shows the concentration of lipid classes in the
whole body of shrimp fed the diets with and without phospholipids supplementation. Total lipid content increased with
the increase in dietary supplemented phospholipids, and
shrimp fed the diet with dietary phospholipids at
101.6 g kg)1 had the highest whole body lipid. Moreover,
dietary phospholipids also influenced the accumulation of
neutral lipid and polar lipid contents in total lipid. The
decrease in free fatty acids (FFA) content directly resulted in
the decrease in neutral lipid content, and the retention of PC
Table 3 The total lipid and lipid class of whole body shrimp fed diets with and without supplementation of phospholipids
One-way
Dietary phospholipids
levels (g kg)1)
P1
27.2
Lipid composition
Total lipid1
Neutral lipid2
TC
TG
FFA
Polar lipid
PC
PE
PI
Others
1.5
42.5
23.7
0.6
17.3
57.5
39.4
12.5
0.29
5.4
P2
36.5
±
±
±
±
±
±
±
±
±
±
0.1b
1.2a
0.4
0.03
0.5a
1.2c
0.6b
0.6
0.02
1.4
1.6
41.1
23.5
0.6
16.3
58.9
40.0
12.4
0.28
6.2
P3
45.8
±
±
±
±
±
±
±
±
±
±
0.1ab
1.6a
0.2
0.01
0.8a
1.6bc
0.8b
0.6
0.03
1.2
1.6
35.9
23.2
0.5
11.7
64.1
43.2
12.3
0.28
8.3
P4
64.4
±
±
±
±
±
±
±
±
±
±
0.1ab
2.5ab
1.0
0.02
1.5b
2.5ab
1.8ab
0.7
0.02
1.2
1.7
35.7
23.7
0.5
10.5
64.3
45.1
13.1
0.28
5.8
P5
101.6
±
±
±
±
±
±
±
±
±
±
0.2ab
1.6ab
0.1
0.01
1.3bc
1.6ab
1.8a
0.1
0.02
0.3
1.9
32.6
23.6
0.5
7.6
67.4
47.1
13.3
0.29
6.7
±
±
±
±
±
±
±
±
±
±
ANOVA
(P-value)
0.1a
1.6b
0.3
0.02
1.3c
1.6a
1.0a
0.2
0.02
0.5
0.027
0.013
0.955
0.349
0.001
0.013
0.008
0.535
0.991
0.368
(ns)
(ns)
(ns)
(ns)
(ns)
Values are shown as means ± SE of three replicates. Values within the same row and not sharing a common superscript are significantly
different (Ducans, P < 0.05).
TC, total cholesterol; TG, triglycerides; FFA, free fatty acids; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol.
1
% Wet weight.
2
% Total lipid.
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Aquaculture Nutrition 17; e615–e621 2010 Blackwell Publishing Ltd
directly resulted in the increase in polar lipid content.
Teshima et al. (1986b) reported that increased total lipid
levels in hepatopancreas and haemolymph in juvenile
P. japonicus as a result of phospholipids supplementation
were caused by an increase in both neutral lipids (triglycerides and cholesterol) and polar lipids (mainly PC). Teshima
et al. (1986a,b,c) indicated that larval P. japonicus feeding on
a diet containing 3% soybean lecithin showed higher whole
body levels of sterol esters (SE), free sterol (FS), PC and PI
compared to larvae fed on a phospholipids deficient diet.
Chen & Jenn (1991) did not observe a preferential increase in
the proportion of PC in total lipids in muscle of P. penicillatus as a result of the supplementation of purified PC up to
50 g kg)1 diet. Chen (1993) observed a preferential increase
in FFA content in muscle of P. monodon as a result of the
increase in dietary phosphatidylcholine. In the work of Gong
et al. (2000a), a higher dietary phospholipids resulted in
higher total lipid in hepatopancreas and lower total lipid in
muscle. In the work of Gong et al. (2000b), increasing purified PC in the diet decreased total lipid, FFA and other polar
lipid levels in hepatopancreas and increased PC in muscle.
The inclusion of phospholipids in the diet affected lipid
deposition of whole body, resulting in increased lipid retention and levels in the animal (Coutteau et al. 1997). In this
experiment, the increased total lipid levels in whole body of
early postlarval L. vannamei as a result of phospholipids
supplementation were found to be the result of increased
polar lipids, mainly PC. Studies using different species
showed different results. The physiological mechanisms
behind this have not been clarified yet. Moreover, the PC
proportion in total lipid did not increase preferentially when
the dietary phospholipids reached 45.8 g kg)1. Thus, it suggests that the early postlarval L. vannamei may require
phospholipids near to 45.8 g kg)1 diet. It is very consistent
with the optimum phospholipids requirement (45.96 g kg)1
diet) by broken-line analysis on weight gain.
Proximate composition of the whole body moisture, crude
protein and lipid seemed to be related to the levels of dietary
phospholipids (Table 4). The contents of crude protein and
lipid increased along with the decrease in whole body moisture. With the increase in the dietary phospholipids level, the
protein content of shrimp in P3, P4 and P5 treatments was
significantly higher than that of shrimp in P1 and P2 treatments, while the moisture content of shrimp in P1 and P2
treatments was significantly higher than that of shrimp in P3,
P4 and P5 treatments. It is well known that phospholipids in
the diet affected lipid deposition, resulting in increased lipid
retention and levels in the animal (Coutteau et al. 1997).
Phospholipids are more polar and may be easily emulsified,
and thus susceptible to a hypothetical bile salt limitation for
their assimilation (Sargent et al. 1993; Coutteau et al. 1997).
Moreover, Olsen et al. (1991) indicated that dietary phospholipids as polar lipid had preferable digestibility compared
with dietary neutral lipid. It can be hypothesized that excessive addition of dietary phospholipids may improve the
digestible energy content. This leads to the reduction in protein as the consumed energy in the diet, so it can be better
synthesized as shrimp body protein, because no decreased
growth was found when the dietary phospholipids level is
increased beyond the required level, the results were similar as
in the previous studies of Chen (1993) and Kanazawa (1993).
Shrimps just like mammals are able to synthesize phospholipids from precursor compounds, such as fatty acids, glycerophosphate, etc. This experiment showed a positive effect
of phospholipids supplementation, and broken-line analysis
on weight gain indicated that the optimum phospholipids
requirement of early L. vannamei postlarvae is 46 g kg)1 diet.
The authors are grateful for the financial support by grant
no. 2007BAD29B04 from the National Key Technology R
and D Program during the 11th five-year plan, China and
Table 4 Whole body composition (g kg)1 wet weight) of shrimp fed diets with and without supplementation of phospholipids
One-way
Dietary phospholipids
levels (g kg)1)
P1
27.2
Whole body composition
Moisture
Protein
Lipid
Ash
822
136
15
39
P2
36.5
±
±
±
±
5a
1b
1b
1
820
136
16
39
P3
45.8
±
±
±
±
6a
1b
1ab
1
775
174
16
40
P4
64.4
±
±
±
±
4c
3a
1ab
1
789
173
17
40
P5
101.6
±
±
±
±
5bc
3a
1ab
1
796
170
19
40
±
±
±
±
ANOVA
(P-value)
6b
2a
1a
1
0.000
0.000
0.027
0.374 (ns)
Values are shown as means ± SE of three replicates. Values within the same row and not sharing a common superscript are significantly
different (Ducans, P < 0.05).
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Aquaculture Nutrition 17; e615–e621 2010 Blackwell Publishing Ltd
co-workers for the help in sampling. We also thank the staff
of the Guangdong Evergreen Group for providing the
experimental base.
AOAC (Association of Official Analytical Chemists) (1984) Official
Methods of Analysis, 14th edn, pp. 114. AOAC, Arlington, VA.
Chen, H.Y. (1993) Requirements of marine shrimp, Penaeus monodon, juvenile for phosphatidylcholine and cholesterol. Aquaculture, 109, 165–176.
Chen, H.Y. & Jenn, J.S. (1991) Combined effects of dietary phosphatidylcholine and cholesterol on the growth, survival and body
lipid composition of marine shrimp, Penaeus peniciliatus. Aquaculture, 96, 167–178.
Conklin, D.E., Devers, K. & Bordner, C.E. (1977) Development of
artificial diets for the lobster, Homarus americanus. Proc. World
Maricult. Soc., 8, 841–852.
Coutteau, P., Camara, M.R. & Sorgeloos, P. (1996) The effect of
different levels and sources of dietary phosphatidylcholine on the
growth, survival, stress resistance, and fatty acid composition of
postlarval Penaeus vannamei. Aquaculture, 147, 261–273.
Coutteau, P., Geurden, I., Camara, M.R., Bergot, P. & Sorgeloos, P.
(1997) Review on the dietary effects of phospholipid in fish and
crustacean larviculture. Aquaculture, 155, 149–164.
Coutteau, P., Kontara, E.K.M. & Sorgeloos, P. (2000) Comparison
of phosphatidylcholine purified from soybean and marine fish role
in the diet of postlarval Penaeus vannamei Boone. Aquaculture,
181, 331–345.
DÕAbramo, L.R., Bordner, C.E. & Conklin, D.E. (1981) Essentiality
of dietary phosphatidylcholine for the survival of juvenile lobsters.
J. Nutr., 111, 425–431.
Folch, J., Lees, M. & Stanley, G.H.S. (1957) A simple method for the
isolation and purification of total lipides from animal tissues.
J. Biol. Chem., 226, 497–509.
Geurden, I., Marion, D., Charlon, N., Coutteau, P. & Bergot, P.
(1998) Comparison of different soybean phospholipidic fractions
as dietary supplements for common carp, Cyprinus carpio, larvae.
Aquaculture, 161, 225–235.
Gong, H., Lawrence, A.L., Jiang, D.H. & Gatlin, D.M. (2000a)
Lipid nutrition of juvenile Litopenaeus vannamei: I. Dietary cholesterol and de-oiled soy lecithin requirements and their interaction. Aquaculture, 190, 305–324.
Gong, H., Lawrence, A.L., Jiang, D.H. & Gatlin, D.M. (2000b)
Lipid nutrition of juvenile Litopenaeus vannamei: I. Active components of soybean lecithin. Aquaculture, 190, 325–342.
Gong, H., Lawrence, A.L., Gatlin, D.M., Jiang, D.H. & Zhang, F.
(2001) Comparison of different types and levels of commercial
soybean lecithin supplemented in semipurified diets for juvenile
Litopenaeus vannamei Boone. Aquac. Nutr., 7, 11–17.
González-Félix, M.L., Lawrence, A.L., Gatlin, D.M. & PerezVelazquez, M. (2002a) Growth, survival and fatty acid composition of juvenile Litopenaeus vannamei fed different oils in the
presence and absence of phospholipid. Aquaculture, 205, 325–343.
González-Félix, M.L., Gatlin, D.M., Lawrence, A.L. & PerezVelazquez, M. (2002b) Effect of dietary phospholipid on essential
fatty acid requirements and tissue lipid composition of Litopenaeus
vannamei juveniles. Aquaculture, 207, 151–167.
Juaneda, A. & Rocquelin, G. (1985) Rapid and convenient separation of phospholipid and non phosphorus lipids from rat heart
using silica cartridges. Lipids, 28, 40–41.
..............................................................................................
Aquaculture Nutrition 17; e615–e621 2010 Blackwell Publishing Ltd
Kanazawa, A. (1993) Essential phospholipid of fish and crustaceans. In: Fish Nutrition in Practice (Kaushik, S.J. & Luquet, P.
eds), pp. 519–530. INRA, Paris. Les Colloques nr. 61, June
24–27, 1993.
Kanazawa, A., Teshima, S., Tokiwa, S., Endo, M. & Abdel Razek,
F.A. (1979) Effect of short-necked clam phospholipid on the
growth of prawn. Bull. Jpn. Soc. Sci. Fish, 45, 961–965.
Kanazawa, A., Teshima, S. & Sakamoto, M. (1985) Effects of dietary
lipids, fatty acids, and phospholipid on growth and survival of
prawn (Penaeus japonicus) larvae. Aquaculture, 50, 39–49.
Kontara, E.K.M., Djunaidah, I.S., Coutteau, P. & Sorgeloos, P.
(1998) Comparison of native, lyso and hydrogenated phosphatidylcholine as source for phospholipids in the diet of postlarval
Penaeus japonicus. Arch. Anim. Nutr., 51, 1–19.
Mu, Y.Y. (2005) De-oiled soy lecithin plays crucial roles in shrimp.
AQUA Culture Asia Pacific, 26–27.
Niu, J., Liu, Y.J., Tian, L.X., Mai, K.S., Yang, H.J., Ye, C.X. &
Gao, W. (2008) Effect of dietary phosphorus sources and varying
levels of supplemental phosphorus on survival, growth and body
composition of larval shrimp (Litopenaeus vannamei). Aquac.
Nutr., 14, 472–479.
Niu, J., Liu, Y.J., Tian, L.X., Mai, K.S., Yang, H.J., Ye, C.X. &
Gao, W. (2009) Nutrient values of dietary ascorbic acid
(L-ascorbyl-2-polyphosphate) on growth, survival and stress tolerance of larval shrimp, Litopenaeus vannamei. Aquac. Nutr., 15,
194–201.
Olsen, R.E., Henderson, R.J. & Pedersen, T. (1991) The influence of
dietary lipid classes on the fatty acid composition of small cod
Gadus morhua L. juvenile reared in an enclosure in northern
Norway. J. Exp. Mar. Biol. Ecol., 148, 59–76.
Paibulkichakul, C., Piyatiratitivorakul, S., Kittakoop, P., Viyakarn,
V., Fast, A.W. & Menasveta, P. (1998) Optimal dietary levels of
lecithin and cholesterol for black tiger prawn Penaeus monodon
larvae and postlarvae. Aquaculture, 167, 273–281.
Piedad-Pascual, F. (1986) Effect of supplemental lecithin and lipid
sources on the growth and survival of Penaeus monodon juveniles.
Asia Fisheries Society. Proceedings of the First Asian Fisheries
Forum, Manila, Philippines, pp. 615–618.
Robbins, K.R., Norton, H.W. & Baker, D.H. (1979) Estimation of
nutrient requirements from growth data. J. Nutr., 109, 1710–1714.
Sargent, J.R., Bell, J.G., Bell, M.V., Henderson, R.J. & Tocher,
D.R. (1993) The metabolism of phospholipids and polyunsaturated fatty acids in fish. In: Aquaculture: Fundamental and
Applied Research, Vol. 43. Coastal and Estuarine Studies (Lahlou,
B. & Vitiello, P. eds), pp. 103–124. American Geophysical Union,
WA.
Shieh, H.S. (1969) The biosynthesis of phospholipids in the lobster,
Homarus americanus. Comp. Biochem. Physiol., 30, 179–184.
Teshima, S., Kanazawa, A. & Kakuta, Y. (1986a) Effects of dietary
phospholipids on lipid transport in the juvenile prawn. Bull. Jpn.
Soc. Sei. Fish., 52, 159–163.
Teshima, S., Kanazawa, A. & Kakuta, Y. (1986b) Role of dietary
phospholipids in the transport of [14C] tripalmitin in the prawn.
Bull. Jpn. Soc. Sci. Fish., 52, 519–524.
Teshima, S., Kanazawa, A. & Kakuta, Y. (1986c) Growth, survival
and body lipid composition of the prawn larvae receiving several
dietary phospholipids. Mem. Fac. Fish., Kagoshima Univ., 35,
17–27.
Thongrod, S. & Boonyaratpalin, M. (1998) Cholesterol and lecithin
requirement of juvenile banana shrimp, Penaeus merguiensis.
Aquaculture, 161, 315–321.
Zeisel, S.H. (1993) Choline deficiency. J. Nutr. Biochem., 1, 332–344.