Optimizing betaine supplementation level in soybean meal-based diets to enhance feed intake and growth performance of juvenile grouper, Epinephelus fuscoguttatus.
The Epinephelinae groupers are popular species for the mariculture industry especially in the Southeast Asian region, due to its high price and demand in the live reef fish trade and seafood industry. Groupers are strict carnivorous species hence it requires feeds with high dietary protein level for optimum growth [1, 2]. Farming of groupers therefore has been relying on the fish meal (FM) based-compounded feeds, as the conventional feeding using low value fish is not sustainable . However, the commercial compounded feeds are getting expensive due to the increased price of FM . Therefore, researches have been carried out to find suitable alternative protein sources to substitute the FM. Plant protein such as soybean meal (SBM) appeared to be a potential alternative protein source because it is generally rich in protein and commercially available in large quantity . In fact, several studies have evidenced that SBM protein can be used to partially replace the FM protein in the diets for juvenile groupers [5, 6, 7]. Although high inclusion level of SBM protein can reduce the diets palatability that causes poor fish intake and growth [5, 6], supplementation of suitable dietary feeding stimulants or feed enhancer (definitions see ) can overcome this problem, as demonstrated in other fish species [9, 10, 11].
In our previous studies, betaine was identified as a feed enhancer  and dietary betaine supplementation indeed can improve the intake of high SBM-based diet in the juvenile grouper, Epinephelus fuscoguttatus . However, the growth performance of the fish was not fully evaluated, and the optimum supplementation level of dietary betaine was not determined. In the present study, we aimed to evaluate the full growth performance of the juvenile grouper fed high SBM-based diets supplemented with betaine and to determine the optimum supplementation level of betaine.
MATERIALS AND METHODS
Formulation of diets:
Formulation of the experimental diets used in the present study is shown in Table 1. A total of five isoproteic (50%) and isolipidic (11%) diets were prepared: (i) Control--FM-based diet, (ii) SBM40 -diet with 40% fish meal protein replaced by dehulled and defatted SBM protein, (iii) BET05, (iv) BET10, and (v) BET15 --diet with 40% of SBM protein supplemented with betaine in the amounts of 0.5%, 1.0% and 1.5% of the diet's total weight, respectively). All the dry ingredients were firstly mixed before the oil and water were added. To supplement betaine into the diets, rightful amount of betaine according to the total weight of the in preparation diet was pre-dissolved into the water added to the mixture of ingredients during mixing. After the mixture became moist dough, it was screw-through a 3 mm die by a meat chopper and the strands were dried at 40 [degrees]C in an oven for about 3 hours. All the experimental diets were refrigerated in 4 [degrees]C until used.
Experimental fish and culture conditions:
The experimental juvenile E. fuscoguttatus (BW 15.6 [+ or -] 0.2 g) were purchased from a local fish farmer. Among all fish, three individuals were randomly sampled for the whole body proximate analysis of initial fish, and another one hundred eighty three individuals were weighed individually and distributed evenly into 15 floating cylinder cages (12 individuals per cage; cage dimension, 50 cm diameter x 50 cm depth) placed randomly in 2 fiberglass tanks (3 tonnes each) that were provided with constant flow-through sea water and aeration. Each treatment of diet was hand-fed to triplicate cages of fish twice daily in morning and afternoon (approximately 0800 and 1400) until apparent satiation level for 8 weeks. The uneaten feed was counted then siphoned out; about 20% of water in each tank was exchanged. The fish were bulk-weighted for every two weeks for monitoring their growth performance; the mortality was recorded every day to calculate the survival rate and feed intake.
Samplings and data analysis:
At the end of the feeding trial, all fish were anesthetized and measured individually for their total length and body weight. Fish that fed on the same diet were pooled and then three individuals were randomly sampled for whole body proximate analysis, and another three were sacrificed for the assessment of hepatosomatic index (HSI) and viscerosomatic index (VSI). Before sampling, the fish were starved for 24 h to assure that their digestive tracts were empty. The fish body weight gain (WG), specific growth rate (SGR), survival rate (SR), feed intake (FI), feed conversion ratio (FCR), net protein utilization (NPU), protein efficiency ratio (PER), condition factor (CF), HSI and VSI were calculated using the following formula:
WG (%) = (Final--Initial fish weight)/ Initial fish weight x 100
SGR (%/d) = [ln (Final weight)--ln (Initial weight)]/ days x 100
SR (%) = (Final fish number/ Initial fish number) x 100
FI (g/ fish) = (Total given feed--Total uneaten feed)/ Fish number
FCR = Dry feed consumed (kg)/ Wet weight gain (kg)
NPU = (Final--Initial fish body protein)/ Total protein intake x 100
PER = Wet weight gain/ Total protein intake
CF = [Fish weight (g)/ Total [length.sup.3]] x 100
HIS = [Liver weight (g)/ Fish weight (g)] x 100
VSI = [Viscera weight (g)/ Fish weight (g)] x 100
Proximate composition analyses:
The proximate composition analyses on the experimental diets and fish whole-body were conducted according to the standard methods by Association of Analytical Chemists . The carcasses of the sampled fish were oven-dried and ground into powder prior to the analysis. The levels of crude protein and lipid were determined following the Kjeldahl method using an automatic system (Kjeltec 2300) and the ether-extraction method in a soxhlet extraction unit (Soxtec 2043), respectively. To determine the ash content, the samples were incinerated at 550[degrees]C in a muffle furnace for 6 hours and weighted the residue remaining. Fiber content of the experimental diets were determined using an semi-auto fiber analysis system (FibertecTM 1020, FOSS) after sample diets were de-fatted using the FibertecTM Cold Extraction Unit 1021.
All data was analyzed using One-Way ANOVA with the Tukey's post hoc test in the SPSS (version 17.0) to check if there were any significant differences among the dietary treatments. Significant differences were assumed when P<0.05.
Proximate composition of formulated diets:
The proximate composition of the formulated diets is shown in Table 2. The analyzed values of the crude protein and lipid were close to the calculated values which are 50% and 11%, respectively. Moisture and ash contents were similar among all the treatments, except that the control treatment contained slightly higher moisture content than the others. Interesting result was found in the crude fiber values. All the SBM-based with betaine--supplemented diets (BET05, BET10, and BET15 treatments) contained about 2 times lower of crude fiber values (6.4-7.6%) than none betaine-supplemented SBM40 treatment (16.9%).
Growth performance and other indices:
The growth performance and feed utilization of fish fed with the experimental diets in the present study are shown in Table 3. As expected, fish fed fish meal-based diet (control diet) attained the best performance in weight gain (WG 171.6%), specific growth rate (SGR 1.7%/ day), and feed intake (FI 47.2 g/fish). Fish fed BET05, BET10, and BET15 diets achieved better WG (124.3, 139.0, and 135.0%, respectively), SGR (1.4, 1.5, and 1.5%, respectively) and FI (32.1, 36.9, and 33.7 g/fish, respectively) than those fed SbM40 diet (WG 100.5%, SGR 1.2 %/day, and FI 25.9 g/fish). The SGR and FI of BET05, BET10, and BET15 were significantly higher than those of the SBM40; however, they were significantly lower than those of the control treatment. The lowest protein efficiency ratio (PER) was found in BET05 (3.6); it was significantly lower than that of the SBM40 treatment (4.5). The condition factor (CF) of the control, BET10 and BET15 treatments were same (1.7), and were significantly higher than that of the SBM40 treatment (1.5). No significant difference was found in the feed conversion ratio (FCR), net protein utilization (NPU), hepato-somatic index (HSI) and viscerasomatic index (VSI) among all treatments. The highest and lowest survival (SR) were recorded in the BET10 (91.7%) and SBM40 (63.9%) treatments, respectively. No significant difference was found among the SR of all treatments.
Proximate composition of fish whole-body:
Table 4 shows the whole-body proximate compositions of fish fed different types of experimental diet. No significant difference was found in the protein, lipid, and ash contents of all treatments. Only moisture content of the control treatment was significantly lower than those of the other treatments.
In the present study, the crude fiber levels of all the betaine-supplemented SBM-based diets were about half of the non-betaine-supplemented diet. This is the first report on such finding; it is hypothesized that the supplementary betaine may flourish the bacteria communities, including those which may help to break down the fiber content in the diets. Such hypothesis was made because betaine is well-known as an osmo-protectant which is essential for bacteria to adapt and survive in stress condition . Further study is necessary to elucidate this hypothesis.
The fish fed betaine-added-SBM diets generally attained better performance in WG, SGR, and FI than the fish fed solely SBM diet, although the results were not comparable to those fed control diet. The FI values of BET05, BET10, and BET15 were all significantly higher than that of the SBM40 treatment. Such results were in parallel with our previous findings that betaine supplementation can be used to enhance the feed intake  although the FI of BET10 in the present study was significantly lower than the control. The SGR values of BET05, BET10, and BET15 were also significantly higher than that of the SBM40 treatment. Apparently, the improvement in FI subsequently enhanced the fish growth because there was no significant difference among the FCR values of all treatments. Supplementation of dietary betaine was also reported to improve the FI and growth performance in the rainbow trout (Oncorhynchus mykiss) fed high SBM-based diets  but not in the largemouth bass (Micropterus salmoides) . Trushenski et al.  also reported that betaine supplementation can improve the FI and growth performance of the juvenile cobia (Rachycentron canadum) fed high soy-derived protein-based diet. These variations clearly demonstrated that the taste effects of betaine can be diverse among different fish species.
The low SR in the SBM40 treatment could be caused by cannibalism led by its very poor FI. The fish vomited most of the feed after capturing them in mouth. Therefore, the starved dominant fish may attack on their weaker mates as fish chasing was frequently observed in the SBM40 treatment cages during feeding, and the found dead fish bodies were generally with many injuries and sometimes missing parts . Besides that, the cannibals could grow bigger in shorter time which would contribute to growth depensation . Therefore, the cannibalism should have also affected the other parameters, beside of survival in the SBM40 treatment. Indeed, the NPU and PER of SBM40 were generally higher than the other treatments, including the control. Such results were abnormal and in contrast with those reported in the previous study . In addition, the moisture, protein and lipid contents in the whole fish body of SBM40 were generally similar to those of the other treatments, indicating that the fish were not in the condition of prolong starvation. These results further supported the positive occurrence of cannibalism in which to compensate the fish growth and survival, despite of the poor feed intake. Apparently, this outcome of the present study supported the suggestion by Hseu et al.  that satiation feeding should be practiced to reduce cannibalism in E. fuscoguttatus, besides of size grading.
Supplementation of dietary betaine is expected to improve the feed utilization as it can flourish the bacteria communities in fish , which should help in the digestion on the alternative source of protein. Substantial studies have been conducted to evaluate the effects of dietary betaine supplementation on fish growth performance [22, 23, 24, 25] but the evaluation on feed utilization was seldom reported. Shankar et al.  reported that dietary betaine supplementation can improve the PER in the Indian major carp (Labeo rohita); however, opposite result was reported by Tiril et al.  in the rainbow trout (O. mykiss). As the PER and NPU in the present study were influenced by cannibalism, evaluation on the effect of betaine on the feed utilization of grouper fed high SBM-based diet is not possible. Further study is required to clarify this theory.
Among all the betaine supplementation diets, fish fed BET10 generally attained the better performance in WG, SGR, and FI than those fed BET05 and BET15, although the results were statistically no significant difference. Therefore, it is concluded that the optimum supplementation level of betaine in the high-SBM based diet for the juvenile grouper is 1.0%.
Corresponding Author: Leong-Seng Lim, Borneo Marine Research Institute, Universiti Malaysia Sabah, 88400 Kota Kinabalu, Sabah, Malaysia.
Received 13 June 2015
Accepted 28 July 2015
Available online 5 August 2015
This study was funded by the Universiti Malaysia Sabah Research Grant Scheme (project code: SBK0036STWN-2012) and partially by the E-Science fund from the Ministry of Science, Technology, and Innovative of Malaysia (MOSTI) (project code: SCF0084-SEA-2012). The authors thank to the management of Tanjung Badak Marine Fish Centre of Sabah Fisheries Department for providing the facilities to conduct the experiment.
 Williams, K.C., 2009. A review of feeding practices and nutritional requirements of postlarval groupers. Aquaculture, 292: 141-152.
 Lim, L.S., A.S.K. Yong and R. Shapawi, 2014. Terrestrial animal- and plant-based ingredients as alternative protein and lipid sources in the diets for juvenile groupers: Current status and future perspectives. Annual Research and Review in Biology, 4: 3071-3086.
 Shapawi, R., S. Mustafa and W.K. Ng, 2011. A comparison of the growth performance and body composition of the humpback grouper, Cromileptes altivelis fed farm-made feeds, commercial feeds or trash fish. Journal of Fisheries and Aquatic Science, 6: 523-534.
 Tacon, A.G.J. and M. Metian, 2008. Global overview on the use of fish meal and fish oil in industrially compounded aquafeeds: Trends and future prospects. Aquaculture, 285: 146-158.
 Luo, Z., Y.J. Liu, K.S. Mai, L.X. Tian, D.H. Liu and X.Y. Tan, 2004. Partial replacement of fish meal by soybean protein in diets for grouper Epinephelus coioides juveniles. Journal of Fisheries of China, 28: 175-181.
 Shapawi, R., I. Ebi and A. Yong, 2013. Soybean meal as a source of protein in formulated diets for tiger grouper, Epinephelus fuscoguttatus juvenile. Part I: Effects on growth, survival, feed utilization and body compositions. Agricultural Sciences, 7: 317-323.
 Shiu, Y.L., S.L. Hsieh, W.C. Guei, Y.T. Tsai, C.H. Chiu and C.H. Liu, In Press. Using Bacillus subtilis E20-fermented soybean meal as replacement for fish meal in the diet of orange-spotted grouper (Epinephelus coioides, Hamilton). Aquaculture Research.
 Kasumyan, A.O. and K.B. Doving, 2003. Taste preferences in fish. Fish and Fisheries, 4: 289-347.
 Dias, J., E.F. Gomes and S.J. Kaushik, 1997. Improvement of feed intake through supplementation with an attractant mix in European seabass fed plant-protein rich diets. Aquatic Living Resources, 10: 385-389.
 Kubitza, F., L.L. Lovshin and R.T. Lovell, 1997. Identification of feed enhancer for juvenile largemouth bassMicropterus salmoides. Aquaculture, 148: 191-200.
 Papatryphon, E. and J.H. Soares Jr., 2000. The effect of dietary feeding stimulants on growth performance of striped bass, Morone saxatilis, fed-a-plant feedstuff-based diet. Aquaculture, 185: 329-338.
 Lim, L.S., W.K. Chor, A.D. Tuzan, R. Shapawi and G. Kawamura, In Press. Betaine is a feed enhancer for juvenile grouper (Epinephelus fuscoguttatus) as determined behaviourally. Journal of Applied Animal Research.
 Lim, L.S., I. Ebi, W.K. Chor, G. Kawamura and R. Shapawi, 2015. Determination on the possibility of dietary betaine supplementation to improve feed intake of soybean meal-based diet in the juvenile grouper (Epinephelusfuscoguttatus): A pilot study. Malaysian Applied Biology, 44: 137-141.
 Association of Official Analytical Chemists (AOAC), 1997. Official methods of analysis of Association of Official Analytical Chemists, International, 16th ed. Arlington, Virginia: AOAC International.
 Csonka, L.N., 1989. Physiological and genetic responses of bacteria to osmotic stress. Microbiological Reviews, 53: 121-147.
 Tiril, S.U., F. Alagil, F.B. Yagci and O. Aral, 2008. Effects of betaine supplementation in plant protein based diets on feed intake and growth performance in rainbow trout (Oncorhynchus mykiss). The Israeli Journal of Aquaculture--Bamidgeh, 60: 57-64.
 Trushenski, J., J. Laporte and H. Lewis, 2011. Fish meal replacement with soy-derived protein in feeds for juvenile cobia: Influence of replacement level and attractant supplementation. Journal of World Aquaculture Society, 42: 435-443.
 Takeshita, A. and K. Soyano, 2009. Effects of fish size and size-grading on cannibalistic mortality in hatchery-reared orange-spotted grouper Epinephelus coioides juveniles. Fisheries Science, 75: 1253-1258.
 Hseu, J.R., P.P. Hwang and Y.Y. Ting, 2004. Morphometric model and laboratory analysis of intracohort cannibalism in giant grouper Epinephelus lanceolatus fry. Fisheries Science, 70: 482-486.
 Hseu, J.R., W.B. Huang and Y.T. Chu, 2007. What causes cannibalization-associated suffocation in cultured brown-marbled grouper, Epinephelus fuscoguttatus (Forsskal, 1775)? Aquaculture Research, 38: 1056-1060.
 He, S.X., Z.G. Zhou, Y.C. Liu, Y.N. Cao, K. Meng, P.J. Shi, B. Yao and E. Ringo, 2012. Do dietary betaine and the antibiotic florfenicol influence the intestinal autochthonous bacterial community in hybrid tilapia (Oreochromis niloticus $ x O. aureus $)? World Journal of Microbiology and Biotechnology, 28: 785-791.
 Normandes, E.B., R.E. Barreto, R.F. Carvalho and H.C. Delicio, 2006. Effects of betaine on the growth of the Fish Piaugu, Leporinus macrocephalus. Brazilian Archives of Biology and Technology, 49: 752-762.
 Luo, Z., X.Y. Tan, X.J. Liu and H. Wen, 2011. Effect of dietary betaine levels on growth performance and hepatic intermediary metabolism of GIFT strain of Nile tilapia Oreochromis niloticus reared in freshwater. Aquaculture Nutrition, 17: 361-367.
 Zakipour Rahimabadi, E., M. Akbari, A. Arshadi and E. Effatpanah, 2012. Effect of different levels of dietary betaine on growth performance, food efficiency and survival rate of pike perch (Sander lucioperca) fingerlings. Iranian Journal of Fisheries Sciences, 11: 902-910.
 Fattahi, S., S.A. Hosseini and M. Sudagar, 2013. The effects of dietary betaine on the growth performance and carcass synthesis of Caspian roach (Rutilus rutiluscaspicus) fingerlings. Scientific Journal of Animal Science, 2: 132-137.
 Shankar, R., H. Shivananda Murthy, Prakash Pavadi and K. Thanuja, 2008. Effect of betaine as feed attractant on growth, survival, and feed utilization in fingerlings of the Indian major carp, Labeo rohita. The Israeli Journal of Aquaculture--Bamidgeh, 60: 95-99.
(1) Leong-Seng Lim, (1) Isabella Ebi, (1) Wei-Kang Chor, (2) Kien Chee Lu, (2) Moris Chong, (2) Ahemad Sade, (1) Rossita Shapawi
(1) Borneo Marine Research Institute, Universiti Malaysia Sabah, Jalan UMS, 88400, Kota Kinabalu, Sabah, Malaysia
(2) Department of Fisheries, Sabah, 4th Floor, Wisma Pertanian Sabah, Jalan Tasik, 88624, Kota Kinabalu, Sabah, Malaysia
Table 1: Formulation of the experimental diets. Ingredient (g per 100 g diet) Diets SBM0 SBM40 BET05 BET10 BET15 Fish meal (a) 62.2 37.3 37.3 37.3 37.3 Soybean meal 0.0 37.2 37.2 37.2 37.2 Tapioca starch (b) 8.6 7.9 7.4 7.4 7.4 Alfa-cellulose 15.0 1.0 1.0 0.5 0.0 CMC (c) 1.5 1.5 1.5 1.5 1.5 Vitamin Premix (d) 3.0 3.0 3.0 3.0 3.0 Mineral Premix (e) 2.0 2.0 2.0 2.0 2.0 Dicalcium phosphate 1.0 1.0 1.0 1.0 1.0 Wheat gluten 3.8 3.8 3.8 3.8 3.8 Fish oil (f) 2.9 5.4 5.4 5.4 5.4 Betaine (g) 0.0 0.0 0.5 1.0 1.5 Total 100.0 100.0 100.0 100.0 100.0 Values in the same row with different superscripts indicate significant difference at level 0.05 (P<0.05) (a) Danish fish meal. (b) Tapioca AAA brand. Bake with Me Sdn. Bhd. (c) Carboxymethyl cellulose (CMC), Sigma. (d) Vitamin mixture (g/kg mixture): ascorbic acid, 45.0; inositol, 5.0; choline chloride, 75.0; niacin, 4.5; riboflavin, 1.0; pyridoxine HCl, 1.0; thiamine HCl,0.92; d-calcium panothenate, 3.0; retinyl acetate, 0.60; vitamin D3, 0.083; Menadione, 1.67; DL alpha tocopherol acetate, 8.0; d-biotin, 0.02; folic acid, 0.09;vitamin B12, 0.00135. All ingredients were diluted with alpha cellulose to 1 kg. (e) Mineral mixture (g/kg mixture): Calcium phosphate monobasic, 270.98; Calcium lactate, 327.0; Ferrous sulphate, 25.0; Magnesium sulphate, 132.0;Potassium chloride, 50.0; Sodium chloride, 60.0; Potassium iodide, 0.15; Copper sulphate, 0.785; Manganese oxide, 0.8; Cobalt carbonate, 1.0; Zinc oxide,3.0; Sodium salenite, 0.011; Calcium carbonate, 129.274. (f) Cod liver oil, Seven Seas Brand. (g) Sigma Brand Table 2: Proximate composition of the experimental diets. Proximate Diets composition (%) Control SBM40 BET05 BET10 BET15 Crude protein 48.9 50.0 50.5 49.4 49.3 Crude lipid 11.0 10.0 11.0 11.0 11.0 Moisture 16.2 11.8 11.5 13.7 12.9 Ash 10.0 9.8 9.9 9.6 9.5 Crude fiber 8.2 16.9 7.6 6.4 6.7 Gross energy 338.1 338.1 338.1 338.1 338.1 (Kcal /100g) Table 3: Growth performance and other indices of fish fed different types of experimental diet. Parameters Diets Control SBM40 WG (%) 171.6 [+ or -] 20.1 (a) 100.5 [+ or -] 7.5 (b) SGR (%/ d) 1.7 [+ or -] 0.1 (a) 1.2 [+ or -] 0.1 (b) FI (g/ fish) 47.2 [+ or -] 3.3 (a) 25.9 [+ or -] 1.1 (b) FCR 2.2 [+ or -] 0.8 (a) 1.9 [+ or -] 0.4 (a) NPU 36.1 [+ or -] 7.7 (a) 44.7 [+ or -] 32.7 (a) PER 4.4 [+ or -] 0.4 (ab) 4.5 [+ or -] 0.3 (a) CF 1.7 [+ or -] 0.0 (a) 1.5 [+ or -] 0.1 (b) HSI 0.01 [+ or -] 0.0 (a) 0.01 [+ or -] 0.0 (a) VSI 0.08 [+ or -] 0.02 (a) 0.07 [+ or -] 0.01 (a) SR (%) 80.6 [+ or -] 17.4 (a) 63.9 [+ or -] 4.8 (a) Parameters BET05 BET10 WG (%) 124.3 [+ or -] 13.8 (b) 139.0 [+ or -] 16.6 (ab) SGR (%/ d) 1.4 [+ or -] 0.1 (c) 1.5 [+ or -] 0.0 (c) FI (g/ fish) 32.1 [+ or -] 2.8 (c) 36.9 [+ or -] 2.1 (c) FCR 1.7 [+ or -] 0.3 (a) 1.9 [+ or -] 0.4 (a) NPU 35.6 [+ or -] 3.4 (a) 36.2 [+ or -] 7.1 (a) PER 3.6 [+ or -] 0.4 (bc) 4.3 [+ or -] 0.1 (ab) CF 1.6 [+ or -] 0.0 (ab) 1.7 [+ or -] 0.0 (a) HSI 0.01 [+ or -] 0.0 (a) 0.01 [+ or -] 0.0 (a) VSI 0.07 [+ or -] 0.0 (a) 0.07 [+ or -] 0.01 (a) SR (%) 83.3 [+ or -] 8.3 (a) 91.7 [+ or -] 0.0 (a) Parameters BET15 WG (%) 135.0 [+ or -] 13.8 (ab) SGR (%/ d) 1.5 [+ or -] 0.0 (c) FI (g/ fish) 33.7 [+ or -] 0.7 (c) FCR 1.9 [+ or -] 0.5 (a) NPU 33.1 [+ or -] 3.2 (a) PER 4.8 [+ or -] 0.1 (a) CF 1.7 [+ or -] 0.0 (a) HSI 0.01 [+ or -] 0.0 (a) VSI 0.07 [+ or -] 0.01 (a) SR (%) 80.6 [+ or -] 12.7 (a) * Different alphabets indicate significant differences at P<0.05 Table 4: Proximate composition of the whole-body of fish fed different types of experimental diet. Proximate Treatments Initial Control composition (%) Crude protein 16.1 [+ or -] 0.6 (a) 18.1 [+ or -] 0.4 (a) Crude lipid 5.5 [+ or -] 0.2 (a) 5.1 [+ or -] 0.3 (ab) Moisture 72.9 [+ or -] 0.8 (a) 70.8 [+ or -] 0.4 (b) Ash 25.3 [+ or -] 0.6 (a) 26.2 [+ or -] 0.2 (a) Proximate SBM40 BET05 composition (%) Crude protein 17.5 [+ or -] 1.1 (a) 18.0 [+ or -] 0.2 (a) Crude lipid 4.1 [+ or -] 0.6 (ab) 3.9 [+ or -] 0.5 (b) Moisture 71.8 [+ or -] 0.1 (a) 71.8 [+ or -] 0.4 (a) Ash 25.0 [+ or -] 0.6 (a) 25.3 [+ or -] 1.0 (a) Proximate BET10 BET15 composition (%) Crude protein 17.4 [+ or -] 0.8 (a) 16.8 [+ or -] 1.2 (a) Crude lipid 4.0 [+ or -] 0.8 (b) 4.0 [+ or -] 0.4 (b) Moisture 72.9 [+ or -] 0.5 (a) 72.2 [+ or -] 1.5 (a) Ash 25.1 [+ or -] 1.2 (a) 25.1 [+ or -] 0.8 (a)
|Printer friendly Cite/link Email Feedback|
|Author:||Lim, Leong-Seng; Ebi, Isabella; Chor, Wei-Kang; Lu, Kien Chee; Chong, Moris; Sade, Ahemad; Shapawi,|
|Publication:||Advances in Environmental Biology|
|Date:||Aug 1, 2015|
|Previous Article:||Corporal punishment: the case of a child abuse by a Malaysian couple in Sweden.|
|Next Article:||Factors influence the larvae distribution and spat settlement of Perna viridis in Marudu Bay, Sabah, Malaysia.|