Mazlan, A.G.

Muchlisin, Z.A.

Muhammadar, A.A.

Samat, A.

Simon, K.D.

Aquatic Commons

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AACL Bioflux, 2011, Volume 4, Issue 4. 
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AACL BIOFLUX 
          Aquaculture, Aquarium, Conservation & Legislation 
                       International Journal of the Bioflux Society 
 
Crude protein and amino acids content in some 
common feeds of tiger grouper (Epinephelus 
fuscoguttatus) juvenile  
1,2Abdullah A. Muhammadar, 1A. Ghaffar Mazlan, 1Abdullah Samat,  
2Zainal A. Muchlisin, and 1K. Das Simon 
 
 
1 School of Environmental and Natural Resource Sciences, Faculty of Science and 
Technology, Universiti Kebangsaan Malaysia, Bangi Selangor D.E., Malaysia; 
2 Department of Aquaculture, Syiah Kuala University, Banda Aceh 2311, Indonesia. 
Corresponding author: M. A. Ghaffar, magfish05@yahoo.com 
 
 
Abstract. The objective of present study was to determine the nutrient contents of natural feed of 
tiger grouper (Epinephelus fuscoguttatus (Forsskål, 1775)) especially for crude protein and amino acids 
content. Five types of feed which have been commonly used by farmers were tested in this study, 
namely shrimp (Mesopodytes sp.), Love Larva® pellet, EP2® pellet, anchovies (Stolephorus spp.) and 
tilapias (Oreochromis mossambicus (Peters, 1852)). The result showed that the average content of 
crude protein were 46.51% in shrimp, 50.16% in pellet Love Larva®, 55.00% in pellet EP2®, 52.54% 
in anchovies and 45.16% in tilapias. The result also showed that EP2® pellet comprised significantly 
higher protein content that other diets (p<0.05). The major essential amino acid in all of five feed 
types were Lysine (2.2 to 3.7%), Leucine (2.5 to 3.6%) and Arginine (2.4 to 3.4 %). The major non 
essential amino acid in all five feed types were Glutamic acid (range from 4.7 to 6.5%) and  Aspartic 
acid (range from 3.1 to 4.7%). Pellet EP2® had highest amino and Aspartic acid compared to other 
diets whereas higher Tyrocine was detected in shrimp.   
Key Words: Aceh Province, type of feeds, crude protein, amino acids content.  
 
 
Introduction. The tiger grouper, Epinephelus fuscoguttatus (Forsskål, 1775) is one of 
the important fishery commodities in Asian countries. It is very popular for seafood, and 
is cultured intensive and extensively in Malaysia, Indonesia and Hongkong (Teng et al 
1978; Liu & Mitcheson 2008; Sugama et al 2008). In Indonesia especially in Aceh 
Province, extensive and semi intensive aquaculture is commonly practice and trash fish is 
usually used as feed item for the fishes. 
The commercial pellets are the most effective feed for grouper because it 
comprises all nutritional components required by the fish. The most common commercial 
feed are EP2® and Love Larva® pellets. However, the pellets are rarely used by farmers 
because its price is relatively high and considered costly for traditional aquaculture or 
extensive and semi-intensive scales. Therefore, the development of mass culture of 
grouper in Indonesia is still infancy.  
In Aceh Province, Indonesia, most farmers used shrimp (Mesopodytes sp.), 
anchovies (Stolephorus spp) and tilapias (Oreochromis mossambicus (Peters, 1852)) as 
alternative feeds to reduce the production cost of grouper. However, the composition of 
nutrients content of these natural feeds were unkown. Moreover, the composition of 
common artificial feeds (EP2®  and Love Larva® pellets) were not reported yet. Therefore, 
in the present study we attempt to evaluate the nutrients content of some natural and 
artificial feeds which have been commonly used for tiger grouper culture with the 
objective of obtaining the best alternative feed for tiger grouper juvenile. 
   
 
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AACL Bioflux, 2011, Volume 4, Issue 4. 
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Material and Method 
  
Sampling. Protein and amino acid profiles of five feeds of grouper (E. fuscoguttatus) i.e. 
shrimp (Mesopodytes sp.), Love Larva® and EP2® pellets, anchovies (Stolephorus spp) 
and tilapias (O. mossambicus) were evaluated. The feeds were collected from the nearby 
farm and transported to laboratory. The feeds were then washed and dried at room 
temperature prior to further analysis. 
Crude Protein and Amino Acids Analysis. Protein content of the dried feeds (shrimp, 
Love Larva® and EP2® pellets, anchovies and tilapias) were determined using Kjeldahl 
method (AOAC 1990). A total of 0.2 grams of powdered samples were added with 6N 
hydrochloric acid (HCl) in borosilicate test tubes. The tubes were covered with parafilem 
and masked tape tightly. The samples were placed in a furnace at temperature of 110°C 
for 24 hours. The samples were then cooled at room temperature for 20 minutes. A total 
of 400 µL of alpha amino butyric acid (AABA) was added into sample and filtered with 
Sartorius stedim filter paper (Biotech, 1288) into 100 mL flask. The samples were then 
diluted with deionized water and then filtered with aqueous syringe nylon filter 0.2 µm. A 
total of 2 mL filtered sample were cooled at -20°C prior to amino acid analysis using 
HPLC standart procedure. 
Data analysis. The data were subjected to one-way analysis of variance (ANOVA), 
followed by comparison of means using Duncan’s multiple range test to determine 
significance of each data (Dytham 2003). All statistical analyses were performed using 
SPSS software v14 (SPSS Inc., Chicago, IL, USA). 
 
Results and Discussion 
 
Crude protein. The ANOVA test showed that the effect of different feed types on the 
crude protein levels were significant (p<0.05). The Duncan multi range test showed that 
the higher crude protein content was found in EP2® pellet and it was significantly different 
than other diets (p<0.05). However, the crude protein content in the shrimp was not 
significantly different than tilapia (p>0.05) (Figure 1).   
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
The average of crude protein content (CP%) of the feeds were 46.51%, 50.16%, 
55.00%, 52.54% and 45.16% for shrimp, Love Larva® pellet, EP2® pellet, anchovies and 
tilapia, respectively. According to Lewell (1989) the optimum protein levels for fish have 
ranged from 25% to 50%. Therefore, in general the crude protein contents in tested 
diets were higher and sufficient for the fish. According to Docan et al (2011) the feed 
0
10
20
30
40
50
60
 Srimph Love Larva ® Pellet  EP2® Pel let Anchovy Tilapia  sp
 Feed types
Cr
ud
e 
Pr
ot
ei
n 
 C
on
te
nt
 (%
)
Shri p Love larvae® pell t 2® p ll t Anchovies Tilapias 
Figure 1. Crude protein content of five feeds for juvenile tiger grouper (different letters 
indicate significant different at p<0.05). 
 
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quality is an important factor to ensure the growth and survival of the cultured fishes. 
Moreover, the carnivorous fishes need higher protein requirements than herbivorous or 
omnivorous. For instance, Malabar grouper (Epinephelus malabaricus (Bloch & Schneider, 
1801)) needs protein with a range from 40% to 50% and 45-52% for other grouper spp. 
(Boonyaratpalin 1997). 
Furthermore, the optimum dietary protein level for E. malabaricus was ranged 
from 40% to 50 (Teng et al 1978). In addition, humpback grouper (Cromileptes altivelis 
(Valenciennes, 1828)) and Malabar grouper (E. malabaricus) required a dietary protein of 
51% and 55%, respectively (Usman et al 2005; Tuan & Williams 2007). Hence, we 
assumed that shrimp, anchovies and tilapia were suitable for alternative diet of tiger 
grouper jouvenile. This is in agreement with Bombeo-Tuburan et al (2001) who reported 
that Oreochromis sp. can be used as a feed for juvenile grouper (Epinephelus coioides 
(Hamilton, 1822)). In addition, Sugama et al (2008) reported that shrimp (Mesopodytes 
sp.) can be consumed effectively by the grouper juvenile. However, further studies are 
needed to evaluate the role of these diets on the survival rate and growth performance of 
tiger grouper. 
Essential amino acids. Essential amino acid profiles of five feeds were showed in Table 
1. The major amino acids in the feeds were Lysine (2.2 to 3.7%), Leucine (2.5 to 3.6%) 
and Arginine (2.4 to 3.4 %). In general, the EP2R has higher essential amino acids level 
compared to other diets. 
The results showed that the alternative feeds tested in this study had nine types 
of essential amino acids. However, the values of essential amino acids were not 
significantly different among the feeds for Arginine, Threonine, Valine, Methionine, 
Isoleucine, Leucine, and Phenylalanine (p>0.05). The higher Histidin and Lysine were 
found in the EP2® pellet. There was significantly difference between EP2® pellet with other 
diets (p<0.05), however, no significantly differences were observed among the EP2® 
pellet shrimps, Love Larva® pellet and anchovies for Lysine (p>0.05), however, these 
values were significantly different than tilapia (p<0.05) (Table 1).   
Actually, fishes require ten types of essential amino acids namely; Arginine, 
Histidine, Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Theorine, Tryptophan 
and Valine (Chau et al 2008; Lewell 1989). However, only nine essesensial amino acids 
were recorded in this study where Tryptophan was not detected by this method. 
However, the level of each essential amino acids requirement are different from species 
to species. For instance, juvenile of red sea bream (Pagrus major (Temminck & Schlegel, 
1843)) requires Histidine, Lysine and Leucine of 12.8 mg-1, 40.6 mg-1 and 41.1 mg-1, 
respectively (Koshio 2002). For Nile tilapia, the Histidine, Lysine and Leucine 
requirements were 1.7%, 5.1% and 2.8-3.6%, respectively (Santiago & Lewell 1988). 
However, the Histidine, Lysine and Leucine requirements for wild juvenile of estuary 
grouper (E. coioides) were 1.62%, 6.49% and 6.16%, respectively (Alava et al 2004; 
Toledo et al 2004).  
Non essential amino acids. The profile of seven non essential amino acids (Aspartic 
acid, Serine, Glutamic acid, Proline, Glycine, Alanine and Tyrosine) from five tested feeds 
(shrimp, Love Larva® and EP2® pellet, anchovies and tilapia) are presented in the Table 2. 
The major non essential amino acid in all feeds were Glutamic acid (ranged from 4.7 to 
6.5%) and Aspartic acid (ranged from 3.1 to 4.7%). The Duncan’s multi range test 
showed that there were no significantly differences among the feeds for Serine, Glutamic 
acid, Proline, Glycine and Alanine (p>0.05). The higher Aspartic acid was found in the 
EP2® pellet, but it was not significantly different than shrimp, Love Larva® pellet and 
anchovies (p>0.05), while the higher Tyrosine was detected in shrimp, but it was not 
significantly different than Love Larva® pellet, EP2® pellet and anchovies (p>0.05). 
Overal, EP2® pellet had higher contents of Aspartic acid, Serine, Glutamic acid, Proline 
and Alanine, while shrimp and anchovies had higher content in Tyrosine and Glycine, 
respectively. 
 
 
 
 
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Table 1 
Percentage of essential amino acid contents in the various alternative feeds of tiger grouper (E. fuscoguttatus).  
Mean of values in the same column followed by a different superscript are significant different (p< 0.05). 
 
Feed HIS ARG THR VAL MET LYS ILE LEU PHE 
A 0.85±0.1a 2.46±0.4a 1.59±0.3a 2.22±0.3a 1.09±0.2a 2.59±0.3ab 2.15±0.3a 3.08±0.4a 1.97±0.3a 
B 1.03±0.2ab 3.19±0.1a 1.87±0.1a 2.19±0.1a 1.37±0.0a 2.94±0.1ab 2.19±0.0a 3.27±0.1a 1.20±0.0a 
C 1.03±0.2ab 3.19±0.1a 1.87±0.1a 2.19±0.1a 1.37±0.0a 2.94±0.1ab 2.19±0.0a 3.27±0.1a 1.20±0.0a 
D 0.96±0.3ab 3.21±1.0a 1.99±0.6a 2.16±0.7a 1.41±0.4a 3.14±0.9ab 1.97±0.6a 3.23±1.0a 1.86±0.6a 
E 0.88±0.0a 2.40±0.1a 1.52±0.1a 1.95±0.1a 1.09±0.0a 2.16±0.2a 1.67±0.1a 2.54±0.2a 1.51±0.0a 
 
Note: A = shrimp, B = Love Larva® pellet, C = EP2® pellet, D = anchovies, E = tilapias, HIS = Histidine, ARG = Arginine, THR = Threonine, VAL = Valine, MET = Methionine, 
LYS = Lysine, ILE = Isoleucine, LEU = Leucine, PHE = Phenylalanine. 
 
 
 
 
Table 2 
Percentage of non essential amino acid contents in the various alternative feeds of tiger grouper (E. fuscoguttatus).  
Mean of values in the same column followed by a different superscript are significant different (p< 0.05). 
 
Feed ASP SER GLU PRO GLY ALA TYR 
A 3.86±0.6ab  1.58±0.3a 5.69±0.7a 1.75±0.4a 2.45±0.3a 2.59±0.3a 1.67±0.2b 
B 4.13±0.1ab 1.82±0.1a 5.67±0.2a 1.90±0.1a 2.16±0.1a 2.40±0.1a 1.59±0.2ab 
C 4.72±0.4b 1.94±0.0a 6.54±0.1a 2.14±0.1a 2.81±0.0a 2.99±0.0a 1.54±0.0ab 
D 3.89±1.1ab 1.93±0.6a 5.87±1.7a 2.10±0.7a 2.89±1.0a 2.65±0.8a 1.53±0.3ab 
E 3.11±0.1a 1.44±0.0a 4.72±0.2a 2.10±0.1a 3.18±0.1a 2.69±0.3a 1.06±0.1a 
 
Note: A = shrimp, B = Love Larva® pellet, C = EP2® pellet, D = anchovies, E = tilapias, ASP = Aspartic acid, SER = Serine, GLU = Glutamic acid, PRO = Proline, GLY = 
Glycine, ALA = Alanine, TYR = Tyrosine.  
 
 
 
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AACL Bioflux, 2011, Volume 4, Issue 4. 
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According to Ghassem et al (2009) Glutamic acid and Aspartic acid were the most 
important essential amino acids for Channa striatus (Bloch, 1793), Pangasianodon 
hypophthalmus (Sauvage, 1878), Clarias batrachus (Linnaeus, 1758) and Oreochromis 
niloticus (Linnaeus, 1758). According to Lewell (1989) insufficient essential amino acids 
in the feed can be replaced by non-essential amino acid. For example, Cysteine can 
replace about 60% of the Methionine. In addition, Tyrosine can replace around 50% of 
the Phenylalanine requirement for channel catfish.  
 
Conclusions. The average content of crude protein contents (CP%) established in the 
feeds were shrimp (46.51%), Love Larva® pellet (50.16%), EP2® pellet (55.00%), 
anchovies (52.54%) and tilapia (45.16%). The EP2® pellet has highest protein content 
and it was significantly different compare to other diets. The major essential amino acid 
in all of the feeds were Lysine (2.2 to 3.7%), Leucine (2.5 to 3.6%) and Arginine (2.4 to 
3.4 %). On average, EP2® pellet had highest amino acid level compared to other diets. 
The major non essential amino acid in the feeds were Glutamic acid (range from 4.7 to 
6.5%) and Aspartic acid (range from 3.1 to 4.7%). In addition, the higher Aspartic acid 
was found in the EP2® pellet, while the higher Tyrosine was detected in shrimp.  
 
Acknowledgements. This study was supported by the Ministry of Science and 
Technology and Innovation Malaysia through Universiti Kebangsaan Malaysia (UKM) 
Science Fund Grant # 04-01-02-SF0124 and UKM Research Grant # UKM-GUP-ASPL-08-
04-235, # UKM-FST-2011. The technical assistance provided by the members of 
Aquaculture Research group of Universiti Kebangsaan Malaysia is acknowledged. 
 
References 
 
Alava V. R., Priolo F. M. P., Arnaiz M., Toledo J. D., 2004 Amino acid and fatty acid 
profiles of wild-sourced grouper (Ephinephelus coides) broodstock and larvae. In: 
Advances in grouper aquaculture. Rimmer M. A., McBridge S., Williams K. C. (eds), 
pp. 47-52. Australian Centre for International Agricultural Research. Canberra.  
AOAC, 1990 [Official methods of analysis of the association of official analytical chemists, 
15th eds. W. Horwitz (ed).] Association of Official Analytical Chemists, Arlington, 
VA, USA, p. 1298. 
Bombeo-Tuburan I., Coniza E. B., Rodriguez E. M., Agbayani R. F., 2001 Culture and 
economics of wild grouper (Epinephelus coioides) using three feed types in ponds. 
Aquaculture 201:229-240. 
Boonyaratpalin M., 1997 Nutritional requirements of marine food fish cultured in 
Southeast Asia. Aquaculture 151:283-313. 
Chau H. Y., Lin Y. H.,  Shiau S. Y., 2008 Nutrition, immunology and health management 
of groupers. In: The aquaculture of grouper. Liao I. C., Leano E. M. (eds). pp. 189-
205, Word Aquaculture Society. USA. 
Docan A., Dediu L., Cristea V., 2011 Effect of feeding with different dietary protein level 
on hematological indices of juvenile Siberian sturgeon, Acipenser baeri reared under 
recirculating systems condition. AACL Bioflux 4(2):180-186. 
Dytham C., 2003 Choosing and using statistics, a biologist’s guide, 2nd edition. Blackwell 
Publishing, Oxford, p. 248. 
Ghassem M., Khoo T. C., Feni H. S., Rozaina T. T. M., 2009 Proximate composition, fatty 
acid and amino acid profiles of selected Malaysian freshwater fish. Malaysian 
Fisheries Journal 8(1):7-16. 
Koshio S., 2002 Red Sea Bream, Pangus majore. In: Nutrient requirements and feeding 
of finfish for aquaculture. CIBI Publishing. New York. p. 418. 
Lewell T., 1989 Nutrition and Feeding of Fish. Van Nostrand Reinhold Publisher, p. 259 
New York. 
Liu M., Mitcheson Y. S. D., 2008 Grouper aquaculture in Mailand China and Hong Kong. 
In: The aquaculture of groupers. Liao I. C., Leano E. M. (eds). pp. 111-142, Word 
Aquaculture Society. USA. 
504 
AACL Bioflux, 2011, Volume 4, Issue 4. 
http://www.bioflux.com.ro/aacl 
Santiago C. B., Lowell R. T., 1988 Amino acid requirements for growth of Nile tilapia. 
Journal of Nutrition 118(12):1540-1546.  
Sugama K., Insan I., Koesshahrani I., 2008 Hatchery and growth-out technology of 
groupers in Indonesia. In: The aquaculture of groupers. Liau I. C., Leano E. M. 
(eds.), p. 341, Word Aquaculture Society. USA. 
Teng S. K., Chua T. E., Lim P. E., 1978 Preliminary observation on the dietary protein 
requirement of estuary grouper, Epinephelus salmoides Maxwell, cultured in floting 
net cages. Aquaculture 15:257-271. 
Toledo J. D., Caberoy N. B., Quinitio G. F., 2004 Environmental factors affecting 
embryonic development, hatching and survival of early stage larvae of the grouper 
(Epinephelus coioides). In: Advances in grouper aquaculture. Rimmer M. A., 
McBridge S., Williams K. C. (eds), pp. 10-16. Australian Centre for International 
Agricultural Research. Canberra.  
Tuan L. A., Williams K. C., 2007 Optimum dietary protein and lipid specifications for 
juvenile Malabar grouper (Epinephelus malabaricus). Aquaculture 267:129-138.  
Usman, Rahmansyah, Lening A., Ahmat T., Williams K., 2005 Optimum dietary protein 
and lipid specifications for grow-out of humpback grouper Cromileptes altivelis 
(Valenciennes). Aquaculture Research 36(13):1285-1292. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Received: 13 May 2011. Accepted: 20 June 2011. Published online: 14 July 2011.   
Authors: 
Abdullah A. Muhammadar, School of Environmental and Natural Resource Sciences, Faculty of Science and 
Technology, Universiti Kebangsaan Malaysia, Bangi Selangor D. E., Malaysia; Department of Aquaculture, Syiah 
Kuala University, Banda Aceh 23111 Indonesia, e-mail: m_dar71@yahoo.com 
A. Ghaffar Mazlan, School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, 
Universiti Kebangsaan Malaysia, Bangi Selangor D. E., Malaysia, e-mail: magfish05@yahoo.com 
Abdullah Samat, School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, 
Universiti Kebangsaan Malaysia, Bangi Selangor D. E., Malaysia, e-mail: nature@ukm.my 
Zainal Abidin Muchlisin, Department of Aquaculture, Syiah Kuala University, Banda Aceh 23111, Indonesia, e-
mail: muchlisinza@yahoo.com 
K. Das Simon, School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, 
Universiti Kebangsaan Malaysia, Bangi Selangor D. E., Malaysia, e-mail: skdas_maa@yahoo.com 
How to cite this article: 
Muhammadar A. A., Mazlan A. G., Samat A., Muchlisin Z. A., Simon K. D., 2011 Crude protein and amino acids 
content in some common feeds of  tiger grouper (Epinephelus fuscoguttatus) juvenile. AACL Bioflux 4(4):499-
504. 
  
 
 
 


Crude protein and amino acids content in some

common feeds of tiger grouper (Epinephelus

fuscoguttatus) juvenile

http://aquaticcommons.org/7528/1/2011_Crude_protein_and_amino_acids_content_in_some.pdf

Crude protein and amino acids content in some common feeds of tiger grouper (Epinephelus fuscoguttatus) juvenile

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