ARTICLE IN PRESS

Comparative Biochemistry and Physiology Part B xx (2002) xxx–xxx

1

3 Dietary amino acid profiles and growth performance in juvenile

4 kuruma prawn Marsupenaeus japonicus
5
1
6 Md. Shah Alam, Shin-ichi Teshima*, Dedy Yaniharto , Manabu Ishikawa, Shunsuke Koshio
7
8 Laboratory of Aquatic Animal Nutrition, Faculty of Fisheries, Kagoshima University, Shimoarata 4-50-20, Kagoshima 890-0056,
Japan 9
10
11 Received 19 March 2002; received in revised form 16 June 2002; accepted 25 June 2002

26 12

27 Abstract
28
29 To assess the reference dietary amino acid profiles for juvenile kuruma prawn Marsupenaeus japonicus a feeding trial
30 was conducted using six semi-purified diets containing casein-gelatin and pre-coated supplemental crystalline amino
31 acids (CAA) and a control diet containing intact protein (casein-gelatin). Pre-coated CAA were supplemented to the
32 diets to simulate dietary amino acid profiles to those of the prawn egg protein (PEP), prawn larvae whole body protein
33 (PLP), prawn juvenile whole body protein (PJP), squid meal protein (SMP), short-necked clam protein (SNP) and
34 brown fish meal protein (BFP). The result showed that kuruma prawn juveniles are capable of utilizing the pre-coated
35 CAA and higher growth performances were observed in the groups fed the PJP, SMP and the control diets than those
36 fed the PLP, SNP, BFP and PEP diets. The essential amino acid proportions (AyE ratios) of the whole body of kuruma
37 prawn differ slightly when compared with the other penaeids or freshwater prawn. The results suggest that PJP and
38 SMP would be suitable as a reference dietary amino acid profile for juvenile prawn.
39 䊚 2002 Elsevier Science Inc. All rights reserved.
40
41 Keywords: Kuruma prawn; Marsupenaeus japonicus; Reference protein; Amino acid profile; Pre-coating; Crystalline amino acid;
42 Utilization; AyE ratio
43

44 52
46 1. Introduction rements of fish and crustaceans are often deter-
mined by feeding experiments with diets 53
47 Since high protein diets are needed for good containing graded levels of the particular amino 54
48 growth of most aquatic animals (NRC, 1993), acid to be examined (Wilson, 1989). However, 55
49 estimation of minimum requirements of essential such a method requires much labor, expense and 56
50 amino acids (EAA) is indispensable to formulate time. In addition, adaptation of the method to 57
51 cost-effective diets. The quantitative EAA requi- slow-eating aquatic animals such as prawns faces 58
the obstacle that supplemental EAA leaches into 59
581 the water before being consumed. Accordingly, 60
582 *Corresponding author. Tel.: q81-99-286-4181; fax: q81- several attempts have been made to assess the 61
583 99-286-4184. EAA requirements based on methods using ideal 62
584 E-mail address: teshima@fish.kagoshima-u.ac.jp
protein and reference amino acid patterns (Boghen 63
585 (S.-i. Teshima).
587 1
Present address: Agency for the Assessment and Applica- et al., 1982; Wilson and Poe, 1985; Castell, 1990; 64
588 tion of Technology (BPPT), Jaluan, M. Husni, Thamrin No. Moon and Gatlin, 1991; Mambrini and Kaushik, 65
589 8; Jakarta Pusat 10340 – Indonesia. 1995). 66

1096-4959/02/$ - see front matter 䊚 2002 Elsevier Science Inc. All rights reserved.
PII: S 1 0 9 6 - 4 9 5 9 Ž 0 2 . 0 0 1 6 5 - 3
 ARTICLE IN PRESS
2 M.S. Alam et al. / Comparative Biochemistry and Physiology Part B xx (2002) xxx–xxx
3 83
Table 1 which were determined by dose-response or radio-
4 Composition of the diets (g per 100 g dry diet) used for juve- isotope methods, the knowledge of EAA require- 84
5
6 nile kuruma prawn
ments of crustaceans are only little and 85

Ingredient CAA-dieta Control fragmentary. 86

12
1
7 1
In a previous study, we have successfully eval- 87
13 Casein 17.0 30.0
14 Gelatin 8.0 15.0 uated the EAA requirements of the Japanese floun- 88
15 Amino acid mixb 20.0 0 der, Paralichthys olivaceus, by using pre-coated 89
16 Cholesterol 1.0 1.0 crystalline amino acids (CAA) which enable retar- 90
17 Squid liver oilc 4.0 4.0 dation of the leaching of CAA into water (Alam 91
18 Soybean lecithind 4.0 4.0 et al., 2002). The kuruma prawn, Marsupenaeus 92
19 Vitamin mixe 7.0 7.0
20 Mineral mixf 4.0 4.0 japonicus, is the most popular penaeid with a high 93

21 a-Starch 8.0 8.0 market value in Japan and many nutritional studies 94
22 Sucrose 7.0 7.0 on this prawn have been conducted (Teshima and 95
23 Glucose 5.0 5.0 Kanazawa, 1971; Teshima et al., 1986, 1989, 2001; 96
24 Carboxymethyl cellulose 4.0 4.0 Shiau, 1998; Ishikawa et al., 2002). However, the 97
25 k-Carrageenan 2.5 2.5
26 a-Cellulose 7.1 7.1 quantitative EAA requirements of this species have 98

Attractantsg 1.4 1.4 not been established. Hence, we intended to eval- 99

27
a
uate the EAA requirements of the prawn by using 100
28 Amino acid patterns of the diets were simulated to those
29 of prawn egg protein (PEP), prawn larvae protein (PLP), the same method as used in the Japanese flounder 101

30 prawn juvenile protein (PJP), squid meal protein (SMP) short- in the viewpoint of comparative nutrition, with 102
31 necked clam protein (SNP) and brown fish meal protein special interest in the difference in EAA require- 103
32 (BFP). ments between fish and prawn. This paper deals 104
b
33 See Table 4. with the relationship between the growth perform- 105
c
34 Feed oil squid (Riken Vitamin, Tokyo, Japan).
35 d
Kanto Chemical Co., Inc., Tokyo, Japan. ance of the prawn, M. japonicus, and dietary amino 106

36 e
(mgy100 g) r-Amino benzoic acid, 47.24; biotin, 1.89; acid profiles by feeding test diets containing pro- 107
37 inositol, 1889.41; nicotinic acid, 188.94; ca-pantothenate, visional reference amino acid profiles. 108
38 283.41; pyridoxine–HCl, 56.70; riboflavin, 37.77; thiamin–
39 HCl, 18.88; menadione, 18.93; vitamin A-palmitate, 47.53; a-
2. Materials and methods 109
40 tocopherol, 94.47; cyanocobalamin, 0.40; calciferol, 5.63;
41 ascorbyl-2-phosphate-Mg, 1474.86; folic acid, 3.79 and choline
42 chloride, 2830.13. 2.1. Experimental diets 110
f
43 (mgy100 g dry diet): MgSO4Ø7H2O, 1422;
44 NaH2PO4Ø2H2O, 370; KH2PO4, 935; Ca(H2PO4)2Ø2H2O, The basal and proximate compositions of the 111
45 1273.
46 g
(gy100 g): Glucosamine–HCl, 0.8; sodium citrate 0.3; CAA-based diets and the control diets are shown 112

47
48 sodium succinate, 0.3. in Tables 1 and 2, respectively. The amino acid 113
profiles of prawn M. japonicus egg protein (PEP), 114
67
prawn larvae (mysis) whole body protein (PLP), 115
The EAA requirements of fish have been con- prawn (50 days after hatching) whole body juve- 116
68 sidered not to vary markedly among species (Kau- nile protein (PJP), squid meal protein (Nippon 117
69 shik, 1998). However, crustaceans such as prawns Suisan Co., Ltd., Japan) (SMP), short-necked clam 118
70 are different to fish in that they have an open- (locally purchased) protein (SNP) and brown fish 119
71 vessel system and contain higher levels of non- meal (Nippon Suisan Co., Ltd., Japan) protein 120
72 protein nitrogen compounds such as free amino 52
73 acids in the body (Dall and Smith, 1987). Amino Table 2
74 acids also play a role in osmotic regulation in The proximate composition of the test diets (% dry matter 53
75 crustaceans (Cheng and Chen, 1998). Considering basis) fed to juvenile kuruma prawn 54
55

76 these facts together, we assume that the EAA Diet 56

77 requirements of crustaceans possibly differ from 57
78 those of fish. Although there are reports on the PEP PLP PJP SMP SNP BFP Control
58
65
64
79 EAA requirements of prawn (Cowey and Forster, Crude protein 42.5 42.0 42.4 42.0 42.6 42.0 43.4 66
80 1971; Shewbart et al., 1972; Kanazawa and Tesh- Total lipid 9.0 9.1 8.6 8.6 9.1 8.6 8.9 67
81 ima, 1981; Smith and Dall, 1991; Chen et al., Ash 5.8 6.1 6.2 6.2 6.2 6.1 6.1 68
Moisture 16.1 16.4 15.6 15.4 15.4 15.6 14.8
82 1992; Fox et al., 1995; Millamena et al., 1996) 70
69
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M.S. Alam et al. / Comparative Biochemistry and Physiology Part B xx (2002) xxx–xxx 3
74 128
Table 3 The other ingredients were according to the recent
75 Amino acid composition (g per 45 g protein) of different ref- nutrient requirement information for juvenile kuru- 129
76
77 erence proteins
ma prawn (Teshima et al., 2001). The amino acid 130

Amino acid PEP PLP PJP SMP SNP BFP Control compositions of the reference proteins, the dietary 131
78
83
82
ingredients, and the supplementation of CAA are 132
84 Arginine 3.14 3.71 4.59 3.72 3.44 2.85 2.49
85 Histidine 1.56 1.01 0.90 1.11 0.96 1.61 0.71 shown in Tables 3 and 4. The preparation of pre- 133
86 Isoleucine 2.66 1.85 1.88 1.99 1.68 1.86 1.76 coated CAA and diets were as per Alam et al. 134
87 Leucine 3.41 2.87 3.19 3.30 2.67 3.40 3.02 (2000). In brief, CAA mixes were pre-coated with 135
88 Lysine 3.66 3.17 3.40 3.48 3.59 4.34 3.48 carboxymethylcellulose (CMC) to prevent leach- 136
89 Methionine 1.07 1.27 1.30 2.14 0.63 1.12 0.91 ing losses. CMC-bound CAA mixture and the 137
90 Phenylalanine 2.08 1.93 1.66 1.91 1.46 2.05 2.17
91 Threonine 2.46 1.79 1.51 1.85 1.61 1.98 1.37 other dry ingredients were added to the casein– 138

92 Tryptophan 0.28 0.65 1.00 0.80 0.38 1.21 0.75 gelatin paste. To improve the water stability of the 139
93 Valine 3.06 2.17 1.88 1.86 1.85 2.21 2.21 diets, gelatinized k-carrageenan was added to the 140
94 Aspartic acid 3.42 4.12 4.14 4.54 4.51 4.25 2.86 mixture. The pH of the diets was adjusted to 7.0– 141
95 Glutamic acid 7.63 6.45 6.76 6.35 6.29 5.44 7.85 7.5 by adding 4 N sodium hydroxide. The pellets 142
96 Serine 1.85 2.00 1.10 1.55 1.85 1.43 1.37
97 Proline 2.05 4.56 3.42 2.78 2.16 2.34 5.24 with 2.1 mm diameter were obtained using a 143

98 Glycine 1.75 2.56 3.38 2.06 2.97 2.40 3.62 pelletizer and dried at 40 8C for 2 h. The dried 144
99 Alanine 2.42 2.16 2.23 2.16 2.55 2.59 2.11 pellets of prawn were steamed at 100 8C for 1 min 145
Tyrosine 1.91 1.78 1.58 1.92 1.18 1.79 2.02 in a cylindrical steamer to improve water stability. 146
101
100
Diets were stored in freezer at y30 8C until used. 147
121
(BFP) were considered as provisional references. 2.2. Experimental prawn and feeding protocol 148
122 Six semi-purified isocaloric diets were formulated
123 to contain the amino acid patterns of PEP, PLP, The brood stock prawn, M. japonicus were 149
124 PJP, SMP, SNP and BFP by supplementing casein– obtained from Matsumoto Suisan, Miyazaki, 150
125 gelatin (approx. 2:1; 25 g) with pre-coated CAA Japan, transported to the Kagoshima University 151
126 (approx. 20 g). A control diet contained only facility, and acclimatized to laboratory conditions. 152
127 intact protein sources (casein and gelatin, 2:1). The eggs were hatched and larvae were reared by 153

105
Table 4
106 Amino acid composition (g per 100 g dry diet) of ingredients and supplementation of CAA in different diets to simulate amino acid
107
108 pattern of references proteins (45% protein)

109 Amino acid Supplied by Supplementation of CAAa

110
111 Casein Gelatin Diet
112
17% 8%
PEP PLP PJP SMP SNP BFP
113
122
121
123 Arginine 0.59 0.78 1.77 2.34 3.22 2.35 2.07 1.48
124 Histidine 0.38 0.02 1.16 0.61 0.50 0.71 0.56 1.21
125 Isoleucine 0.82 0.17 1.67 0.86 0.89 1.00 0.69 0.87
126 Leucine 1.40 0.29 1.72 1.18 1.50 1.61 0.98 1.71
127 Lysine 1.53 0.41 1.72 1.23 1.46 1.54 1.65 2.40
128 Methionine 0.45 0.06 0.56 0.76 0.79 1.63 0.12 0.61
129 Phenylalanine 1.01 0.21 0.86 0.71 0.44 0.69 0.24 0.83
130 Threonine 0.61 0.16 1.69 1.02 0.74 1.08 0.84 1.21
131 Tryptophan 0.42 0.01 0 0.22 0.57 0.37 0 0.78
132 Valine 1.01 0.23 1.82 0.93 0.64 0.62 0.61 0.97
133 Aspartic acid 1.06 0.53 1.83 2.53 2.55 2.95 2.92 2.66
134 Glutamic acid 3.42 0.97 3.24 2.06 2.37 1.96 1.90 1.05
135 Serine 0.56 0.20 1.09 1.24 0.34 0.79 1.09 0.67
136 Proline 1.63 1.26 0 1.67 0.53 0 0 0
137 Glycine 0.22 1.73 0 0.61 1.43 0.11 1.02 0.45
138 Alanine 0.41 0.74 1.27 1.01 1.08 1.01 1.40 1.44
Tyrosine 1.06 0.09 0.76 0.63 0.43 0.77 0.03 0.64
139
a
140
141 Supplemented as L-form (Ajinomoto Co., Inc., Japan).
 ARTICLE IN PRESS
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145
Table 5
146
147 Growth performance of the juvenile kuruma prawn fed test diets for 40 days

Diet Weight gain (%) FCEa PERb APRc Survival (%)

148
153
152
154 PEP 131.3"4.7a 0.26"0.02 0.62"0.02a 9.4"0.3a 78"4
155 PLP 168.0"4.6b 0.34"0.06 0.81"0.03bc 10.9"0.2ab 78"4
156 PJP 206.7"4.7c 0.35"0.02 0.96"0.05d 12.6"0.9b 73"7
157 SMP 210.0"3.5c 0.33"0.03 0.97"0.02d 13.2"0.7b 75"4
158 SNP 170.7"3.5b 0.26"0.02 0.70"0.02ab 8.5"0.4a 80"0
159 BFP 184.0"4.6b 0.30"0.03 0.86"0.03cd 10.5"0.64ab 80"0
Control 212.7"5.5c 0.34"0.04 0.92"0.04cd 10.9"0.6ab 78"4
160
161 Values are means"S.E. of triplicate groups. Means with different letter in the same column differ significantly (P-0.05).
a
162 Feed conversion efficiency (FCE)sweight gain (g)ytotal feed intake in dry weight basis (g).
b
163 Protein efficiency ratio (PER)sweight gain (g)ytotal protein intake in dry weight basis (g).
c
164
165 Apparent protein retention (APR)sprotein gain=100yprotein intake.
154 189
feeding live food (Ceatocerus and Artemia) and sively washed with 7% TCA, ethanol, chloroformy
155 commercial diets (Higashimaru Feed, Shrimp seed methanol (3:1), and diethyl ether followed by 190
156 production number 0-1, Kagoshima, Japan) for 2.5 centrifugation to collect precipitates. Amino acid 191
157 months. Before starting the feeding trial, the juve- analyses of protein fractions, ingredients of the test 192
158 niles were sorted to obtain equal size prawns and diets and whole body of the juveniles were con- 193
159 transferred into other tanks. ducted according to Teshima et al. (1986) using 194
160 Seven triplicate groups of prawns, 0.50"0.01 g high performance liquid chromatography (HPLC), 195
161 (mean"S.D.) in initial body weight, were fed the (Shimadzu LC-6A, Japan). Ash and moisture con- 196
162 respective test diets in 21 rectangular tanks with a tents were analyzed by standard methods (AOAC, 197
163 plastic cover (30 l capacity, filled with 28 l sea 1990). Crude lipid was determined by the method 198
164 water) for 40 days. The tanks were individually of Bligh and Dyer (1959). Crude protein was 199
165 equipped with a false bottom and sand substrate determined by the Kjeldahl method (total- 200
166 that acted to filter the water. Water was circulated N=6.25) with a Tecator Kjeltec System (1007) 201
167 at 0.8 lymin through the action of an air-lift device. Digestion system, 1002 Distilling unit, and Titra- 202
168 Fifteen prawns were stocked randomly in each tion unit, Sweden) using boric acid to trap released 203
169 tank and diets were given daily by hand at 8–10% ammonia. 204
170 of body weight at 08:30 and 16:30 h. Known
171 quantities of diets were given, and fecal matter 2.4. Statistical analysis 205
172 and uneaten diets were collected by siphoning
173 from the tank every morning. Lighting was provid- All data were subjected to statistical verification 206
174 ed using a 12 h light:12 h dark regime. Body using one-way analysis of variance (package 207
175 weight was measured every 10 days of feeding super-ANOVA, ver. 1.11, Abacus Concepts, Berke- 208
176 periods when the tanks and sand were cleaned and ley, California, USA). Significant differences 209
177 filled with seawater again. Water temperature, between means were evaluated by the Tukey– 210
178 salinity and pH were monitored daily. The mean Kramer test (Kramer, 1956). Probabilities of P- 211
179 values of water temperature, salinity and pH value 0.05 were considered significant. 212
180 during the experimental period were 23.5"1.5 8C,
181 33.5"0.71 ppt and 8.02"1.8, respectively. These 3. Results 213
182 values were thought to be suitable environmental
183 conditions for the prawn M. japonicus (Shigueno, Table 5 presents the mean weight gain, percent 214
184 1975). survival, feed conversion efficiency (FCE), protein 215
efficiency ratio (PER) and apparent protein reten- 216
185 2.3. Biochemical analysis tion (APR) of the juveniles fed test diets. Percent 217
survivals were not significantly affected among 218
186 The protein fraction of dietary protein sources the dietary treatments. The highest weight gain 219
187 was obtained by homogenizing wet ingredients was obtained for prawn fed the control, PJP and 220
188 with 10% trichloroacetic acid (TCA) and succes- SMP diets, whereas the lowest weight gain was 221
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169
Table 6
170 Body composition (% dry matter basis) (means"S.E.) of kuruma prawn fed test diets. Means with different letters in the same column
171
172 differ significantly (P-0.05)

Diet Dry matter Crude protein Total lipid Ash

173
178
177
179 PEP 23.2"0.5c 58.1"0.2a 9.8"0.2b 23.2"0.2c
180 PLP 21.8"0.2abc 59.5"0.1b 9.4"0.1ab 21.9"0.4ab
181 PJP 22.2"0.6abc 59.1"0.1b 9.3"0.1ab 22.4"0.4bc
182 SMP 22.6"0.5bc 58.5"0.1ab 9.8" 0.1b 21.0"0.3a
183 SNP 20.7"0.2ab 58.1"0.2a 9.3"0.2ab 22.4"0.1bc
184 BFP 21.0"0.5ab 58.1"0.2a 9.4"0.1ab 22.4"0.1bc
Control 20.2"0.4a 59.4"0.3b 8.8"0.1a 21.8"0.1ab
186
185

222 238
found in prawn fed the PEP diet. Thus the weight shown in Table 7. Some differences in the level
223 gains for the prawn fed the PEP, PLP, SNP and of a few amino acids, such as arginine, histidine 239
224 BFP diets were significantly lower than those of and proline were significant (P-0.05) among the 240
225 the PJP, SMP and control diets. There was no dietary groups. The prawn fed the PJP and SNP 241
226 significant difference in FCE between the different diets had higher arginine levels than these fed the 242
227 groups. The SMP, PJP, BFP and control diets gave BFP diet, but no significant differences with those 243
228 statistically higher PER than the PEP and SNP fed the PEP, PLP, SMP or the control diets. Higher 244
229 diets. Protein, lipid and ash contents (% dry matter histidine was found in prawn fed the PJP and SNP 245
230 basis) of the whole body after growth trial were diets than in those fed the other test diets. The 246
231 influenced by the different patterns of the dietary other whole body amino acid concentrations did 247
232 amino acid in the diets (Table 6). Significantly not show any significant differences among the 248
233 higher protein contents were observed from the dietary treatments. 249
234 prawn fed the PLP, PJP and control diets than the The AyE ratio w(each EAA contentytotal EAA 250
235 other diets. content including cystine and tyrosine)=1000x of 251
236 The amino acid compositions of the whole body the EAA composition of the whole body after the 252
237 of the prawn after 40 days of the feeding trial are growth trial are shown in Table 8. The AyE ratios 253

190
Table 7
191 Amino acid composition (g per 100 g dry sample) of the whole body of the juvenile kuruma prawn fed diets with different amino acid
192
193 profiles

194 Amino acids Diet

195
PEP PLP PJP SMP SNP BFP Control
196
203
202
204 Arginine 3.49ab 3.56ab 3.69b 3.21ab 3.77b 2.72a 3.41ab
205 Histidine 1.07a 1.18a 1.48b 1.14a 1.45b 1.00a 1.31ab
206 Isoleucine 1.23 1.40 1.24 1.32 1.38 1.20 1.21
207 Leucine 2.38 2.63 2.33 2.40 2.56 2.13 2.30
208 Lysine 2.70 3.11 2.77 2.71 2.95 2.42 2.83
209 Methionine 0.76 0.93 0.73 0.75 0.84 0.72 0.69
210 Phenylalanine 2.22 2.14 2.03 1.99 2.24 1.82 1.90
211 Threonine 1.38 1.50 1.35 1.41 1.42 1.24 1.31
212 Tryptophan 0.50 0.60 0.63 0.42 0.53 0.40 0.34
213 Valine 1.49 1.40 1.31 1.39 1.39 1.33 1.32
214 Aspartic acid 3.33 3.60 3.36 3.35 3.55 3.00 3.37
215 Glutamic acid 6.40 6.96 6.25 6.44 6.77 5.82 6.50
216 Serine 1.32 1.39 1.22 1.27 1.33 1.15 1.31
217 Proline 2.50ab 2.75b 2.26ab 2.62ab 2.30ab 1.87a 2.16a
218 Glycine 3.97 4.71 3.57 3.60 4.62 3.77 3.84
219 Alanine 2.24 2.56 2.21 2.11 2.08 1.81 2.36
220 Tyrosine 2.18 2.01 2.25 1.98 2.15 2.01 1.91
Taurine 1.08 1.01 0.95 0.86 1.06 0.76 1.11
221
222
223 Values are means of triplicate groups.
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227
Table 8
228 AyE ratiosa of the essential amino acid composition of whole body tissue of the kuruma prawn fed different reference amino acid
229
230 profile diets and compare with P. monodon and M. rosenbergii

231 Diet P. M.
PEP PLP PJP SMP SNP BFP Control monodonb rosenbergiic
232
238
237
239 Arg 180 174 186 171 182 160 184 153 145
240 His 55 58 75 61 70 59 71 47 68
241 Ile 63 68 63 71 67 71 65 85 73
242 Leu 123 129 118 128 124 125 124 146 147
243 Lys 139 152 140 145 143 143 153 145 164
244 Met 39 46 37 40 41 42 37 74d 70
245 Phee 227 203 216 212 212 225 206 155 172
246 Thr 71 73 68 75 69 73 71 76 88
247 Trp 26 29 32 22 26 24 18 16 ND
Val 77 68 66 74 67 78 71 100 72
248
249 N.D.snot detected.
a
250 AyE ratios(each EAA contenty total EAA content including cystine and tyrosine)=1000.
b
251 Dy-Penaflorida (1989).
c
252 Reed and D’Abramo (1989).
d
253 Methionine and cystine.
e
254
255 Phenylalanine and tyrosine.
254 283
among the dietary groups did not show much tion of CAA by prawn, as also observed in our
255 variation except for arginine (ranging from 160 to previous study on flatfish Japanese flounder (Alam 284
256 186), histidine (from 55 to 75) and tryptophan et al., 2000). In fish and crustacean, feeding of 285
257 (from 18 to 32). The AyE ratios of the whole dietary CAA results in rapid increase in plasma 286
258 body were compared to the previously reported amino acid concentrations compared with that of 287
259 values for penaeid shrimp P. monodon (Dy-Pena- intact dietary protein (Deshimaru, 1976; Dabrows- 288
260 florida, 1989) and freshwater prawn M. rosenbergii ki, 1983). From the present study, we assume that 289
261 (Reed and D’Abramo, 1989) (Table 8). pre-coated CAA not only reduces the leaching 290
losses but also makes CAA absorption slowly in 291
262 4. Discussion the digestive tract of the prawn after ingestion. 292
Thus supplementation of CAA with coatings can 293

263 Earlier, attempts to investigate the utilization of be an efficient means for the estimation of EAA 294

264 CAA by replacing dietary protein on kuruma requirements of prawn as also reported for fish 295

265 prawn (Deshimaru and Kuroki, 1974) and tiger like rainbow trout (Cho et al., 1992). 296

266 prawn (Pascual, 1989) were unsuccessful due to In the present study, the body weight gain of 297

267 leaching of CAA from the diets. CAA supplemen- kuruma prawn fed the diet with amino acid profile 298

268 tation in the diet of P. vannamei yielded positive of prawn juvenile whole body protein (PJP) was 299

269 results, but it was still inferior to the control diet significantly higher than those fed the PEP, PLP, 300

270 (Lim, 1993). Millamena et al. (1996) have suc- SNP and BFP diets. This result supported the 301

271 ceeded in preparing water stable CAA based diet suggestion of Reed and D’Abramo (1989) that the 302

272 using CMC and k-carrageenan as binders to deter- whole body or tail muscle amino acid profile could 303
273 mine the amino acid requirements of tiger prawn, be used as reference dietary profiles for freshwater 304
274 P. monodon. In the present study, the weight gain prawn M. rosenbergii. In the case of fish, the EAA 305
275 on the SMP and PJP diets containing a mixture of pattern of the whole body of a given species has 306
276 casein-gelatin and pre-coated CAA were not sig- been considered to be representative of the EAA 307
277 nificantly different with that on the control diet requirement profile of that species (Wilson and 308
278 containing intact protein. This result showed that Cowey, 1985; Mambrini and Kaushik, 1995). In 309
279 kuruma prawn juveniles were capable of utilizing contrast, the amino acid profiles of whole hen egg 310
280 pre-coated CAA. It therefore appears that coating (Halver et al., 1959) and fish eggs (Ketola, 1982) 311
281 of CAA before mixing all ingredients in the diet have been suggested to reflect the EAA require- 312
282 is an applicable technique to improve the utiliza- ments of fish. Deshimaru and Shigueno (1972) 313
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314 366
also reported the best growth for kuruma prawn trout (Kaushik and Luquet, 1980). Dall and Smith
315 fed a squid meal-based diet. Deshimaru (1982) (1987) also showed that concentration of whole 367
316 proposed that the proportional composition of ami- muscle total amino acids in tiger prawn P. esculen- 368
317 no acids found in the whole body of short-necked tus changed only slightly when starved, but con- 369
318 clam could be ideal for kuruma prawn. In the centrations of various free amino acids changed 370
319 present study, however, the diets with the amino significantly. 371
320 acid profiles of the prawn egg (PEP) and short- Several studies have assessed the requirements 372
321 necked clam (SNP) gave significantly lower of EAA using the concept of AyE ratio (Moon 373
322 weight gain than those with amino acid profiles of and Gatlin, 1991). The concept of AyE ratio was 374
323 squid meal (SMP) and prawn juveniles proteins first introduced by Arai (1981) to formulate test 375
324 (PJP). There is much information to show that diets for coho salmon Oncorhynchus kisutch. In 376
325 well-balanced amino acid profiles in the diet are the present research, the calculated AyE ratios of 377
326 needed for good growth of the prawn (Millamena the whole body of kuruma prawn for arginine and 378
327 et al., 1997) and carp (Murai et al., 1989). We phenylalanine were a little higher as compared 379
328 deduce that the amino acid profiles of squid meal with the reported value for tiger shrimp P. mono- 380
329 protein and prawn juvenile whole body protein don (Dy-Penaflorida, 1989). However, the AyE 381
330 might reflect the requirement pattern of EAA for ratio of leucine for the kuruma prawn was a little 382
331 the prawn. lower than those values reported for P. monodon 383
332 In the present study, the amino acid profiles of and M. rosenbergii (Reed and D’Abramo, 1989). 384
333 the juvenile prawn protein and squid meal protein The other AyE ratios in the whole body did not 385
334 are thought to be ideal amino acid patterns for show a big difference as compared with other tiger 386
335 good growth of M. japonicus juveniles not only shrimp except methionine (as cystine content was 387
336 on the basis of weight gain data but also consid- not determined). This suggests that the differences 388
337 ering on the values of PER and APR. The values in the requirement values for EAA between M. 389
338 of these parameters are similar to those of the high japonicus and P. monodon could be minor, as 390
339 quality diet (Giri et al., 1997) and higher than pointed out by D’Abramo (1998) from a compar- 391
340 those of the other diets with the amino acid profiles ison of AyE ratios between M. rosenbergii and 392
341 of PEP and SNP diet (Table 5). The differences penaeid species. 393
342 in protein retention among the groups could be In conclusion, juvenile prawn whole body pro- 394
343 due to bioavailability of dietary amino acids by tein as well as squid meal protein appears to 395
344 prawn. Therefore, the amino acid profile of the provide the most appropriate dietary amino acid 396
345 juvenile prawn protein (PJP) or the squid meal profiles for the maximum growth of juvenile kuru- 397
346 protein (SMP) appeared to better reflect reference ma prawn. These data will provide information to 398
347 dietary amino acid patterns for juvenile kuruma prepare balanced amino acid based diets for kuru- 399
348 prawn. ma prawn until the requirements of all EAA are 400
349 The concentration of amino acids in a certain established. 401
350 protein in animal bodies is commonly regarded as
351 fixed by the messenger RNA sequence. In fact, Acknowledgments 402
352
Kaushik (1998) did not find any significant dif- The scholarship received by Md. Shah Alam 403
353 ference in the total whole body amino acids of from the Ministry of Education, Culture, Sports, 404
354 two different sizes of European sea bass, gilthead Science and Technology (Monbukagakusho) of 405
355 sea bream and turbot. In the present study, the Japan and the scholarship for D. Yaniharto from 406
356 whole body total amino acid composition (% dry Science and Technology for Industrial Develop- 407
357 sample) of the juvenile prawn, was not altered by ment Project, Indonesia are gratefully acknowl- 408
358 dietary treatments, except for some amino acids as edged. The authors wish to acknowledge 409
359 also observed in fish such as rainbow trout Ajinomoto Co., Inc., Japan for supplying crystal- 410
360 (Mohanty and Kaushik, 1991) and Japanese floun- line amino acids. 411
361 der (Alam et al., 2002). The small differences of
362 certain amino acid levels in the whole body among References 412
363 the dietary treatments found in the present study Alam, M.S., Teshima, S., Ishikawa, M., Koshio, S., 2000. 413
364 may be due to the differences in the tissue levels Methionine requirement of juvenile Japanese flounder Par- 414
365 of free amino acids as also observed in rainbow alichthys olivaceus. J. World Aquacult. Soc. 31, 618–626. 415
 ARTICLE IN PRESS
8 M.S. Alam et al. / Comparative Biochemistry and Physiology Part B xx (2002) xxx–xxx
416 476
Alam, M.S., Teshima, S., Yaniharto, D., Ishikawa, M., Koshio, Fox, J.M., Lawrence, A.L., Li-Chan, E., 1995. Dietary require-
417 S., 2002. Influence of different dietary amino acid pattern ment for lysine by juvenile Penaeus vannamei using intact 477
418 on growth and body composition of juvenile Japanese and free amino acid sources. Aquaculture 131, 279–290. 478
419 flounder Paralichthys olivaceus. Aquaculture 210, 359–369. Giri, I N.N., Teshima, S., Kanazawa, A., Ishikawa, M., 1997. 479
420 AOAC, 1990. In: Helric, K. (Ed.), Official Methods of Effects of dietary purodoxine and protein levels on growth, 480
421 Analysis of AOAC. 15th ed. Association of Official Ana- vitamin B6 content, and free amino acid profile of juvenile 481
422 lytical Chemists Inc, Arlington, VA,. Penaeus japonicus. Aquaculture 157, 263–275. 482
423 Arai, S., 1981. A purified test diet for coho salmon, Oncor- Halver, J.E., Delong, D.C., Mertz, E.T., 1959. Methionine and 483
424 hynchus kisutch, fry. Nippon Suisan Gakkaishi 47, 547–550. cystine requirements of chinook salmon. Fed. Proc. 18, 484
425 Boghen, A.D., Castell, J.D., Conklin, D.E., 1982. In search of 2076. 485
426 a reference protein to replace ‘vitamin free casein’ in lobster Ishikawa, M., Teshima, S., Koshio, S., Kanazawa, A., 2002. 486
427 nutrition studies. Can. J. Zool. 60, 2033–2038. Movement of triolin and cholestane in the digestive tract of 487
428 Bligh, E.G., Dyer, W.J., 1959. A rapid method of total lipid prawn Marsupenaeus japonicus. Aquacult. Res. 32, 488
429 extraction and purification. Can. J. Biochem. Physiol. 37, 375–382. 489
430 911–917. Kanazawa, A., Teshima, S., 1981. Essential amino acids of the 490
431 Castell, J.D., 1990. Reference diet for crustacean principles of prawn. Bull. Jpn. Soc. Sci. Fish. 47, 1375–1377. 491
432 experimentation.Advances in Tropical Aquaculture. Actes Kaushik, S.J., 1998. Whole body amino acid composition of 492
433 des Colloques 9, IFREMER, Plouzane, France. pp. 339–354. European sea bass (Dicentrarchus labrax), gilthead sea 493
434 Chen, H.Y., Len, Y.T., Roelants, I., 1992. Quantification of bream (Sparus aurata) and turbot (Psetta maxima) with an 494
435 arginine requirements of juvenile marine shrimp Penaeus estimation of their IAA requirement profiles. Aquat. Living 495
436 monodon using microencapsulated arginine. Mar. Biol. 114, Resour. 11, 355–358. 496
437 229–233. Kaushik, S.J., Luquet, P., 1980. Influence of dietary amino 497
438 Cheng, S.Y., Chen, J.C., 1998. Effects of nitrite exposure on acid patterns on the free amino acid contents of blood and 498
439 the hemolymph electrolyte, respiratory protein and free muscle of rainbow trout (Salmo gairdnerii R.). Comp. 499
440 amino acid levels and water content of P. japonicus. Aquatic Biochem. Physiol. 64B, 187–190. 500
441 Toxicol. 44, 129–139. Ketola, H.G., 1982. Amino acid nutrition of fishes: require- 501
442 Cho, C.Y., Kaushik, S.J., Woodward, B., 1992. Dietary arginine ments and supplementation of diets. Comp. Biochem. Phy- 502
443 requirement of young rainbow trout (Oncorhynchus mykiss). siol. B 73, 17–24. 503
444 Comp. Biochem. Physiol. 102A, 211–216. Kramer, C.Y., 1956. Extension of multiple range tests to group 504
445 Cowey, C.B., Forster, J.R.M., 1971. The essential amino acid means with unequal number of replications. Biometrics 12, 505
446 requirements of the prawn, Palaemon serratus. The growth 307–310. 506
447 of prawns on diets containing proteins of different amino Lim, C., 1993. Effect of dietary pH on amino acid utilization 507
448 acid composition. Mar. Biol. 10, 77–81. by shrimp (Peneaus vannamei). Aquaculture 114, 293–303. 508
449 D’Abramo, L.R., 1998. Nutritional requirements of the fresh- Mambrini, M., Kaushik, S.J., 1995. Indispensable amino acid 509
450 water prawn Macrobrachium rosenbergii: comparisons with requirements of fish: correspondence between quantitative 510
451 species of penaeid shrimp. Rev. Fish. Sci. 6, 153–163. data and amino acid profiles of tissue proteins. J. Appl. 511
452 Dabrowski, K., 1983. Comparative aspects of protein digestion Ichthyol. 11, 240–247. 512
453 and amino acid absorption in fish and other animals. Comp. Millamena, O.M., Bautista-Teruel, M.N., Kanazawa, A., 1996. 513
454 Biochem. Physiol. A 74, 417–425. Methionine requirements of juvenile tiger shrimp Penaeus 514
455 Dall, W., Smith, D.M., 1987. Changes in protein-bound and monodon Fabricus. Aquaculture 143, 403–410. 515
456 free amino acids in the muscle of the tiger prawn Penaeus Millamena, O.M., Bautista-Teruel, M.N., Reyes, O.S., Kana- 516
457 esculentus during starvation. Mar. Biol. 95, 509–520. zawa, A., 1997. Threonine requirement of juvenile marine 517
458 Deshimaru, O., 1976. Studies on a purified diet for prawn. 6. shrimp Penaeus monodon. Aquaculture 151, 9–14. 518
459 Absorption rate of amino acids in amino acid test diet. Bull. Mohanty, S., Kaushik, S.J., 1991. Whole body amino acid 519
460 Jpn. Soc. Sci. Fish. 42, 331–335. composition of Indian major carps and its significance. 520
461 Deshimaru, O., 1982. Protein and amino acid nutrition of the Aquat. Living Resour. 4, 61–64. 521
462 prawn Penaeus japonicus. In: Proceedings of the 2nd Inter- Moon, H.Y., Gatlin, D.M., 1991. Total sulfur amino acid 522
463 national Conference on Aquaculture Nutrition. Pruder, G.D., requirement of juvenile red drum, Sciaenops ocellatus. 523
464 Landgon, C., Conklin, D. (Eds.). Louisiana State University, Aquaculture 95, 97–106. 524
465 Baton Rouge, LA, pp. 106–123. Murai, T., Daozun, W., Ogata, H., 1989. Supplementation of 525
466 Deshimaru, O., Shigueno, K., 1972. Introduction to the artifi- methionine to soy flour diets of fingerling carp Cyprinus 526
467 cial diet for prawn Penaeus japonicus. Aquaculture 1, carpio. Aquaculture 77, 373–385. 527
468 115–133. NRC (National Research Council), 1993. Nutrient Require- 528
469 Deshimaru, O., Kuroki, K., 1974. Studies on a purified diet ments of Warmwater Fishes and Shellfishes. National Acad- 529
470 for prawn. III. A feeding experiment with amino acid test emy Press, Washington, DC p. 102. 530
471 diets. Bull. Jpn. Soc. Sci. Fish. 41, 101–103. Pascual, F.P., 1989. Status of shrimp nutrition and feed devel- 531
472 Dy-Penaflorida, V., 1989. An evaluation of indigenous protein opment in Southeast Asia. In: De Silver, S.S. (Ed.), Pro- 532
473 sources as potential component in the diet formulation for ceedings of the Third Asian Fish Nutrition Network 533
474 tiger prawn, Penaeus monodon, using essential amino acid Meeting, Asian Fisheries Society Special Publication 4, 534
475 index (EAAI). Aquaculture 83, 319–330. Asian Fisheries Society, Manila, Philippines. pp. 80–89. 535
 ARTICLE IN PRESS
M.S. Alam et al. / Comparative Biochemistry and Physiology Part B xx (2002) xxx–xxx 9
536 557
Reed, L., D’Abramo, L.R., 1989. A standard reference diet for larvae of the prawn, Penaeus japonicus. Aquaculture 51,
537 crustacean nutrition research. 3. Effects on weight gain and 225–235. 558
538 amino acid composition of whole body and tail muscle of Teshima, S., Kanazawa, A., Koshio, S., Horinouchi, K., 1989. 559
539 juvenile prawns Macrobrachium rosenbergii. J. World Aqua- Lipid metabolism of the prawn Penaeus japonicus during 560
540 cult. Soc. 20, 107–113. maturation: variation in lipid profiles of the ovary and 561
541 Shewbart, K.L., Meis, W.L., Ludwig, P.D., 1972. Identification hepatopancreas. Comp. Biochem. Physiol. B 92, 45–49. 562
542 and quantitative analysis of the amino acids present in Teshima, S., Koshio, S., Ishikawa, M., Kanazawa, A., 2001. 563
543 protein of the brown shrimp, Penaeus aztecus. Mar. Biol. Protein requirement of the prawn Marsupenaeus japonicus 564
544 16, 64–67. estimated by a factorial method. Hydrobiologia 449, 565
545 Shigueno, K., 1975. Shrimp Culture in Japan. Association for
293–300. 566
546 International Technical Promotion, Tokyo p. 153.
Wilson, R.P., 1989. Amino acids and proteins. In: Halver, J.E. 567
547 Shiau, S.Y., 1998. Nutrient requirements of penaeid shrimp.
548 Aquaculture 164, 77–93. (Ed.), Fish Nutrition. second ed. Academic Press, New 568

549 Smith, D.M., Dall, W., 1991. Metabolism of proline by the York, pp. 112–153. 569
550 tiger prawn Penaeus esculentus. Mar. Biol. 110, 85–91. Wilson, R.P., Cowey, C.B., 1985. Amino acid composition of 570
551 Teshima, S., Kanazawa, A., 1971. Biosynthesis of sterols in whole body tissue of rainbow trout and Atlantic Salmon. 571
552 the lobster Panulirus japonicus, the prawn Penaeus japoni- Aquaculture 48, 373–376. 572
553 cus, and the crab Portunus trituberculatus. Comp. Biochem. Wilson, R.P., Poe, W.E., 1985. Relationship of whole body 573
554 Physiol. B 38, 597–602. and egg essential amino acid patterns to amino acid require- 574
555 Teshima, S., Kanazawa, A., Yamashita, M., 1986. Dietary ment patterns in channel catfish, Ictalurus punctatus. Comp. 575
556 value of several proteins and supplemental amino acids for Biochem. Physiol. B 80, 385–388. 576
577