Growth, body composition, and ammonia tolerance of juvenile white shrimp Litopenaeus vannamei fed diets containing different carbohydrate levels at low salinity.
KEY WORDS: shrimp, Litopenaeus vannamei. low salinity, carbohydrate, ammonia tolerance, diet, growth
The white shrimp Litopenaeus vannamei is the largest shrimp species in aquaculture, accounting for more than 70% of the world's farmed shrimp (Li et al. 2009) production. With the development of inland low-salinity shrimp farming, L. vannamei has become one of the most important shrimp species for aquaculture in many countries (Saoud et al. 2003, Cheng et al. 2006). The shrimp L. vannamei is euryhaline and can tolerate a wide range of salinity, from 1-50 (Pante 1990), but ambient low-salinity water can affect its growth performance and physiological status (Kinne 1971). At salinities less than 5, slow growth, low survival rates, and poor stress tolerance were found in L. vannamei (Diaz et al. 2001, Lin & Chen 2001, Lin & Chen 2003, Li et al. 2007, Li et al. 2008); however, the culture of white shrimp at low salinities has continued to increase throughout the world during the past decade (Roy et al. 2010). Therefore, there is a need to explore further the physiological status of white shrimp cultured at low salinities and to identify practical ways to improve the growth performance and physiological adaptation of L. vannamei to low salinity.
Although attempts have been made, through dietary manipulation, to improve the physiological status of Litopenaeus vannamei at low salinities, little success has been achieved. Increase of dietary highly unsaturated fatty acids did not change the ion regulation at low salinity (Hurtado et al. 2007). Inclusion of cholesterol and lecithin did not improve the growth performance of L. vannamei at a low salinity (Roy et al. 2006). In contrast, a high-protein diet improved the growth of L. vannamei at a low salinity because the dietary amino acids served as osmotic regulators to reduce the energy loss from muscle and therefore promote the somatic growth of shrimp (Cuzon et al. 2004, Li et al. 2011). However, more dietary protein means greater feed costs, which do not contribute to the sustainable development of this industry. Because osmoregulation is an energy-cost process, carbohydrate (CBH) metabolism as a cheaper energy source should be considered in shrimp diet formulation. Besides, CBH is the primary and immediate source of energy for shrimp (Lehninger 1978), and can meet the high energy demands of aquatic animals existing under stressful conditions (Welcomme & Devos 1991, Tseng & Hwang 2008, Wang et al. 2012). When challenged with salinity, blood glucose levels increased in Chasmagnathus granulatus (Santos & Nery 1987) and Crangon crangon Linnaeus (Spaargaren & Mors 1985). Chloride ion in seawater could promote gluconeogenesis to satisfy the high energy requirement under salinity stress for Lihinia emarginata and Charybdis natator, as indicated by the increased glucose-6-phosphatase activity (Scholnick et al. 2006). Comparison of the growth and metabolism of L. vannamei fed high and low CBH diets with low and high protein contents at salinities of 15 and 40 showed that a low-CBH diet activates the gluconeogenic pathway, by which shrimp convert protein to glycogen to meet their energy demands (Rosas et al. 2001, Rosas et al. 2002). As far as we know, however, no attempt has been made to study the effects of CBH level on L. vannamei juveniles at salinities less than 5--an environment in which most inland shrimp are farmed.
Therefore, this study examined the roles of different levels of diary CBHs on growth performance, body composition, key enzyme activities in glycometabolism, and ammonia tolerances of juvenile Litopenaeus vannamei at a low salinity of 3. Results obtained from this study would provide a practical solution, through dietary manipulation, to improve growth performance and to reduce environmental stress for inland white shrimp farming at low salinities.
MATERIALS AND METHODS
Six isonitrogenous (40%) and isolipid (6%) diets, using fishmeal as protein, and fish oil and soybean oil (1:1) as lipid sources, were formulated to contain 5%, 10%, 15%, 20%, 25%, and 30% of corn CBH (Table 1). Dietary ingredients were ground by 80-size mesh and weighed accurately on an electronic balance. All dry ingredients were mixed thoroughly before lipids and water were added. Diets were processed into 3-mm-diameter pellets, dried at room temperature to a moisture content less than 100 g/kg diet, ground and sieved to an appropriate size, and stored at -20[degrees]C until use (Peres et al. 2003).
Experimental Animals and Facilities
Juvenile Litopenaeus vannamei were obtained from a local farm in Hainan, China. Shrimp were cultured in fiberglass tanks (80 X 60 X 50 cm) at a salinity of 32 for 1 wk, and were then acclimated to the target salinity of 3 by changing 2.0 per day. During the acclimation, shrimp were fed with a commercial feed containing 42% crude protein and 9.5% crude lipid. Seawater was pumped from the sea at a depth of 10 m close to the Baisha coast near Haikou City. Tap water was aerated before being added to the tank to adjust water salinity. Daily water exchange was around one third of the tank volume. Water quality parameters were monitored 2-3 times a week throughout the feeding trial, and were maintained at 26.8-28.8[degrees]C, 6.63-7.34 mg dissolved oxygen/L, and less than 0.06 mg total ammonia nitrogen/L.
Experiment Design and Sampling
After acclimation to a salinity decrease from 32 to 3, 30 Litopenaeus vannamei juveniles (8.6 [+ or -] 0.5 mg) in triplicate tanks (40 X 50 X 50 cm) were assigned randomly to each of the experimental diets and were fed twice daily (0800 hr and 1700 HR) for 42 days. Based on the amount of feed left from the previous day, daily rations were adjusted to a slight oversatiation. The uneaten feed was removed daily with a siphon tube. At the end of the experiment, all shrimp in each tank were counted and starved for 24 h before being weighed. The growth performance and morphometric indexes were calculated as follows:
Weight gain (%) = 100 x [[W.sub.t] - [W.sub.0]/[W.sub.0]]
where [W.sub.0] is the initial weight and [W.sub.t] is the final weight.
Survival rate (%) = 100 x [Final number of shrimp/Initial number of shrimp],
CF = 100 x [W/[L.sup.3]]
where CF is the condition factor, W is shrimp wet weight in grams, and L is the whole body length in centimeters.
HSI (%) = 100 x Hepatopancreas weight/Body weight.
where HIS is the hepatosomatic index.
Biochemical Composition Analysis
All experimental diets and fish samples were analyzed in triplicate for proximate composition following standard methods (AOAC 2000). Moisture was determined by oven drying at 105[degrees]C to a constant weight. Samples used for dry matter determination were digested with nitric acid and incinerated in a muffle furnace at 600[degrees]C overnight for ash weight. Protein was measured by the combustion method using an FP-528 nitrogen analyzer (Leco Corporation, St. Joseph, MI). Lipid was determined by the ether extraction method using the Soxtec system (2055 Soxtec Avanti; Foss Tecator, Hoganas, Sweden).
Soluble Protein and Glycogen Content Determination
The hepatopancreas was homogenized (1:10 w/v) in ice-cold potassium phosphate buffer (137 mmol/L NaCl, 2.7 mmol/L KC1, 4.3 mmol/L Na2HP04, 1.4 mmol/L K[H.sub.2]P[O.sub.4], pH 7.4). Soluble protein in the hepatopancreas was measured with the Lowry method using serum bovine albumin as a standard (Lowry et al. 1951). The glycogen concentrations in the hepatopancreas and muscle homogenates were measured as follows (Dubois et al. 1956). Homogenate (100 mg/mL, w/v) was prepared in 5% trichloroacetic acid buffer for 2 minutes at 8,000g\ After collection, the glycogen was dissolved by the addition of 0.5 mL distilled water, and then 5 mL concentrated sulfuric acid and phenol (5%) were added and mixed. The glycogen content was read at 490 nm on a spectrophotometer and the glycogen concentration was expressed as milligrams glycogen per gram wet weight.
Enzyme Activities Assays
Pyruvate kinase activity was determined as follows (Feksa et al. 2003). Samples of the hepatopancreas from different treatments were homogenized (1:10 w/v) in ice-cold buffer with a homogenizer. Pyruvate kinase activity was assayed in an incubation medium containing 0.1 M Tris-HCl buffer, 10 mM Mg[Cl.sub.2], 0.16 mM NADH, 75 mM KC1, 5.0 mM ADP, 1.0 U L-lactate dehydrogenase, and 10 pL of the mitochondria-free supernatant at pH 7.5 in a final volume. The reaction was started using 1.0 mM phosphoenolpyruvate. All assays were performed in duplicate at 25[degrees]C. Results are expressed as units per milligram protein.
Glucose-6-phosphate dehydrogenase (G6PDH) activity was assayed as follows (Nagayama et al. 1972). The hepatopancreas samples from each tank were homogenized (1:10 w/v) in ice-cold buffer (0.15 M KOH, 0.004 M MgS04, 0.004 M EDTA, and 0.004 M acetylcysteine). Glucose-6-phosphate dehydrogenase activity was assayed in the incubation medium containing 0.2 M Mg[Cl.sub.2], 0.025 M G6P, 0.0075 M NDAP, and distilled water. After 3-5 min of incubation, G6PDH activity was estimated by measuring the concentration of NADPH at 340 nm at 20[degrees]C. Results are expressed as units per milligram protein.
Unless otherwise stated, all preparations were performed at 0-4[degrees]C. A homogenate (1:10 w/v) was prepared in 0.25 M buffered sucrose (0.05 M potassium phosphate buffer at pH 7.4, 10 mM [beta]-mercaptoethanol, and 0.25 M sucrose) using a homogenizer (Mishra & Shukla 2003). The absorbance at 340 nm of oxidation-reduction reactions catalyzed by malate dehydrogenase reduced over time. Enzyme activity was calculated by the change of absorbance in 1 min. Results are expressed as units per milligram protein. All protein contents in the samples were measured by the method of Lowry et al. (1951), and serum bovine albumin was used as a standard.
Ammonia Nitrogen Tolerance
Nine shrimp remaining in each tank were used for the challenge with 9.33 mg/L ammonium nitrogen at salinity of 3 for 96 h without feeding (Li et al. 2007). The testing medium was refreshed every 24 h at 26.8-27.5[degrees]C (pH 8.0 [+ or -] 0.2). Shrimp mortality was recorded at 6 h, 12 h, 24 h, 48 h, 72 h, and 96 h.
To compare the effects of dietary treatments on all the parameters tested, and the survival rate at each time point in the trial testing ammonia nitrogen tolerance, data were subjected to 1-way analysis of variance (SPSS 19.0). We converted the variance, which was not neat before statistical analysis, to guarantee all data were tested on homogeneity of variance. If a significant difference was identified for the diet treatment, the differences among all diet levels were compared using Duncan's multiple range tests. A significance level of P < 0.05 was used for all statistical tests.
Weight gain and survival rate of shrimp fed 20% CBH were the greatest in all groups, and significantly greater than shrimps fed 5% CBH and~30% CBH (Table 2). Shrimp fed 25% CBH had the greatest HSI, which differed from that of shrimp fed 5%-20% CBH. No significant differences were found in condition factor among all treatments (Table 2).
Whole-body crude protein and lipid contents tended to increase with elevated dietary CBH levels (Table 3). Crude protein content of shrimp fed 25% CBH and 30% CBH was significantly greater than shrimp fed 5%-20% CBH (Table 3). Crude body lipid levels of shrimp fed 20%-30% CBH were significantly greater than those fed 5% CBH. No significant differences were found in moisture and ash content.
Hepatopancreas soluble protein increased with the increase of dietary CBH and peaked in shrimp fed 20% CBH, which was significantly greater than that of shrimp fed 5% CBH and 10% CBH. No significant differences in hepatopancreas soluble protein levels were found among shrimp fed the 15%, 25%, and 30% CBH diets (Table 4). Both hepatopancreas glycogen and muscle glycogen were significantly affected by dietary CBH levels, showing a pattern of up and down with an increase of dietary CBH. Hepatopancreas glycogen levels of shrimp fed 15% CBH and 20% CBH were significantly greater than other groups. Shrimp fed 15% CBH had the greatest muscle glycogen content, which differed significantly from that of shrimp fed 25% CBH and 30% CBH (Table 4). Glucose-6-phosphate dehydrogenase activity also showed a pattern of up and down with an increase of dietary CBH (Table 4). Shrimp fed 15% CBH had the greatest G6PDH activity, which was significantly different from those fed 5%, 10%, 20%, and 30% CBH. No significant differences were found in pyruvate kinase and malate dehydrogenase among all treatments.
After the 96-h ammonia nitrogen challenge, the survival rate of the shrimp fed 20% CBH was the greatest (92.21%). The lowest survival rate occurred in shrimp fed 25% CBH. The survival rates of the shrimp fed 25% and 30% CBH were lower than those fed 5%-20% CBH. However, no significant differences in shrimp survival rate were observed among CBH levels throughout the experiment (Fig. 1).
As an osmoregulator, Litopenaeus vannamei has a strong ability to regulate its osmotic pressure and blood ions in a certain level when exposed to an ambient salinity change, during which they need much more extra energy (Pante 1990, Pequeux 1995). Therefore, as a cheaper, primary, and immediate source of energy for organisms in stress conditions (Lehninger 1978, Welcomme & Devos 1991, Tseng & Hwang 2008, Wang et al. 2012), CBH nutrition may play an essential role in the osmoregulation of L. vannamei because CBH is often included in shrimp diet (Alava & Pascual 1987, Shiau et al. 1991, Shiau & Peng 1992, Cruz-Suarez et al. 1994, Cuzon et al. 2004). In this study, shrimp fed 20% CBH had the best growth performance and the greatest survival rate. Greater dietary CBH inclusions decreased the growth of L. vannamei, which is in agreement with previous finding that excess dietary CBH can reduce diet digestibility in aquatic animals, in part, because digestive enzymes for CBH are relatively low compared with CBH in the substrate (Stone 2003). In addition, a-amylase activity was saturated in Litopenaeus stylirostris fed 21% CBH, indicating that the digestibility of CBH in this species is limited by the [alpha]-amylase enzyme substrate balance (Rosas et al. 2000). For L. vannamei cultured at an ambient salinity of 6-14, 15% dietary CBH is optimal for its growth (Guo et al. 2011). In contrast, this study showed that more dietary CBH (20%) is needed to satisfy the extra energy demand for optimal growth at a low salinity of 3 for juvenile L. vannamei. Similarly, L. vannamei at salinity of 15 used more dietary CBH as an energy source than those at a salinity of 40 (Rosas et al. 2000). Therefore, more dietary CBH content contributes to energy provided for L. vannamei to deal with hypoosmotic stress through osmoregulation and ion regulation at low salinity (Welcomme & Devos 1991. Rosas et al. 2000, Tseng & Hwang 2008, Wang et al. 2012), meanwhile guaranteeing the normal growth of white shrimp.
In this study, whole-body crude protein and lipid levels, and hepatopancreas soluble protein content were improved by an elevated level of dietary CBH. Higher whole-body crude protein and hepatopancreas soluble protein content can be explained by the protein-sparing effect of dietary CBH (Rosas et al. 2001), because Litopenaeus vannamei need more protein for growth or as a source of metabolic energy to ensure sufficient free amino acids for osmoregulation at low salinity (Li et al. 2009, Li et al. 2011), and at this time the dietary CBH can replace the protein or amino acids as a source of energy. Besides, excess CBH can be modified and stored as lipids in animals in the form of triacylglycerides or sterols, and most CBH appears to be directed toward storage as glycogen and lipids rather than crossing directly to the hemolymph (Verri et al. 2001), which explains the greater whole-body crude lipid storage in this study. Shrimp need more energy to adapt to stress and more hemolymph proteins as a source of osmotic effectors or as metabolic energy at low salinities, which leads to the slow growth of L. vannamei. Therefore, the findings in this study indicate that enough dietary CBH can improve the slow growth of shrimp at low salinities. Therefore, the protein-sparing effect and increased lipid storage associated with increasing dietary CBH may explain why the shrimp fed 25% CBH and 30% CBH had a greater HS1 than those fed 5%-20% CBH, because the hepatopancreas is not only the main site for the secretion of digestive enzymes (Oliveira & Da Silva 1997), but also it is the primary site for nutrient reserve (Al-Mohanna & Nott 1987).
In this study, a tendency of increased glycogen and soluble protein in the hepatopancreas and muscle glycogen, followed by a decline with an increase of dietary CBH was observed, which is possibly related the glycogen synthesis affected by the soluble protein content, because Litopenaeus vannamei can convert protein to glycogen through gluconeogenesis, depending on the degree to which the amino acid is used for osmotic regulation at low salinities (Rosas et al. 2001). We found that the glycogen content decreased in shrimp fed a greater level of CBHs. Because shrimp are able to store glycogen in muscle and the hepatopancreas, the glycogen content does not continue to increase in muscle and the hepatopancreas with an increase of dietary CBH. Similarly, there was a saturation curve of digestive gland glycogen concentration in relation to dietary CBH in Litopenaeus stylirostris (Rosas et al. 2000).
In this study, malate dehydrogenase, pyruvate kinase, and G6PDH dehydrogenase activity was not affected by dietary CBH levels, indicating that the glycolytic pathway and pentose phosphate pathway are not affected by the level of dietary CBH in Litopenaeus vannamei at low salinities. This can be explained in part by the saturation of digestive enzyme activity. The results from Chasmagnathus granulatus are in contrast to our results, as the pyruvate kinase level in the jaw muscle of crabs fed a high-CBH diet was 4-fold greater than in crabs fed a high-protein but low-CBH diet (Marqueze et al. 2006). In European sea bass and gibel carp, hepatic pyruvate kinase activity increased when fish were fed diets with a high starch content (Enes et al. 2006, Tan et al. 2009). Moreover, there was no effect of diet composition on hepatopancreas glucose uptake and glycogen synthesis (Marqueze et al. 2011). In previous studies, CBH metabolism, such as crustacean hyperglycemic hormone, is involved in the osmoregulation of aquatic animals (Charmantier et al. 1988, Spanings-Pierrot et al. 2000, Morris 2001). Currently, the role of CBH in osmoregulation is not fully understood and needs further investigation.
White shrimp are more susceptible to ambient toxicants at low salinities, including ammonia (Lin & Chen 2001, Li et al. 2007), boron (Li et al. 2008), nickel (Leonard et al. 2011), and pesticides (Wang et al. 2013), because the optimal salinity for the growth of Litopenaeus vannamei is around 20 (Li et al. 2007). When L. vannamei is exposed to a diluted medium or low salinities, extremely low salinities pose a serious stress on L. vannamei, and more energy is needed to improve the osmoregulation capacity to release the adverse effects. Shrimp fed 20% CBH had the greatest survival after they were challenged with ammonia nitrogen for 96 h, indicating that under hypo-osmotic stress, a moderate level of CBHs can serve as a direct energy source for osmoregulation (Welcomme & Devos 1991, Tseng & Hwang 2008). Because the survival rate of shrimp fed 20% CBH was only greater numerically and did not differ significantly from shrimp in other treatments, further studies should be conducted to confirm this finding.
Overall, 15%-20% CBH in the diet of Litopenaues vannamei improved growth performance at inland low-salinity ponds by providing direct extra energy for growth and osmoregulation. A dietary CBH level of 20% alleviated the adverse effect of ammonia stress at a low salinity, but additional studies are needed to understand the mechanism why dietary CBH may relieve the stress of ammonia in the white shrimp.
This project was supported by grants from National Natural Science Foundation of China (nos. 31172422, 31001098, and 31472291), the Special Fund for Agro-Scientific Research in the Public Interest (nos. 201003020 and 201203065), the National Key Technology Support Program (no. 2012BAD25B03), the Shanghai Committee of Science and Technology (no. 10JC1404100), the Specialized Research Fund for the Doctoral Program of Higher Education of China (no. 20100076120006), and in part by the Innovation fund from East China Normal University and the E-Institute of Shanghai Municipal Education Commission (no. E03009). We thank Qiu Yong from Hainan University for his help during the experiment.
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XIAODAN WANG, (1) ERCHAO LI, (1) * JIAN G. QIN, (2) SHIFENG WANG, (3) XUEFEN CHEN, (3) YAN CAI, (3) KE CHEN, (1) YINGMEI HOU, (3) NA YU, (1) MEILING ZHANG, (1) ZHENYU DU (1) AND LIQIAO CHEN (1) *
(1) Laboratory of Aquaculture Nutrition and Environmental Health, School of Life Sciences, East China Normal University, No. 500 Dongchuan Road, Shanghai, 200062, China; (2) School of Biological Sciences, Flinders University, Adelaide, SA 5001, Australia; (3) Department of Aquaculture, Hainan University, No. 58, Renmin Road, Haikou, Hainan, 570228, China
* Corresponding author. E-mail: firstname.lastname@example.org; lqchen@bio. ecnu.edu.cn
TABLE 1. Percentage composition and analyzed nutrient contents of experimental diets. Diets g/100 g Ingredients 1 2 3 Fish meal 55.0 55.0 55.0 Corn starch 5.0 10.0 15.0 Fish oil 2.0 2.0 2.0 Soybean oil 2.0 2.0 2.0 Lecithin 1.0 1.0 1.0 Cholesterol 0.5 0.5 0.5 Vitamin Premix * 2.0 2.0 2.0 Mineral Premix ([dagger]) 0.5 0.5 0.5 Carboxymethyl cellulose 2.0 2.0 2.0 Cellulose 28.5 23.5 18.5 Calcium carbonate 1.5 1.5 1.5 Crude protein 40.19 40.49 40.81 Crude lipid 5.90 5.99 6.17 Diets g/100 g Ingredients 4 5 6 Fish meal 55.0 55.0 55.0 Corn starch 20.0 25.0 30.0 Fish oil 2.0 2.0 2.0 Soybean oil 2.0 2.0 2.0 Lecithin 1.0 1.0 1.0 Cholesterol 0.5 0.5 0.5 Vitamin Premix * 2.0 2.0 2.0 Mineral Premix ([dagger]) 0.5 0.5 0.5 Carboxymethyl cellulose 2.0 2.0 2.0 Cellulose 13.5 8.5 3.5 Calcium carbonate 1.5 1.5 1.5 Crude protein 39.94 39.96 40.33 Crude lipid 6.01 5.98 6.12 * Vitamin premix, diluted in cellulose, contained the following vitamins (measured in milligrams per kilogram diet): vitamin A, 350,000 IU; vitamin D3,450,00 IU; vitamin E, 20 g; menadione, 7.5 g; thiamin, 10 g; riboflavin, 10 g; pyridoxamine, 12 g; cobalamin, 20 mg; nicotinamide, 40 mg; folic acid, 3 g; calcium pantothenate, 30 g; biotin, 100 mg; ascorbic acid, 60 g; and inositol, 60 g. ([dagger]) Trace mineral premix contained the following minerals (measured in grains per 100 g premix): cobalt chloride, 0.004: cupric sulfate pentahy-drate, 0.250: ferrous sulfate, 4.0: magnesium sulfate heptahydrate, 28.398; manganous sulfate monohydrate, 0.650: potassium iodide, 0.067: sodium selenite, 0.010: zinc sulfate heptahydrate, 13.193: sodium dihydrogen phosphate, 15: and filler, 38.428. TABLE 2. Growth performance and morphological parameters of Litopenaeus vannamei fed experimental diets for 42 days. CBH level (%) Weight Survival gain (%) rate (%) 5 5,808.56 (ab) 63.33 (a) 10 4,871.06 (a) 76.67 (bc) 15 6,161.96 (bc) 80 (c) 20 7,097.92 (c) 81.11 (c) 25 6,704.78 (bc) 65.55 (ab) 30 4,955.94 (a) 66.67 (ab) Pooled SEM 370.68 4.08 CBH level (%) Hepatosomatic Condition index (%) factor (%) 5 3.12 (a) 1.14 10 3.24 (a) 1.20 15 3.2 (a) 1.21 20 3.41 (a) 1.14 25 5.02 (b) 1.25 30 4.02 (ab) 1.26 Pooled SEM 0.43 0.09 The different superscripts of the same column values are significantly different (P < 0.05). CBH. carbohydrate. TABLE 3. Whole-body proximate composition as wet weight of Litope- naeus vannamei fed different experimental diets for 42 days. CBH level Moisture Crude protein Crude lipid Ash (%) (%) (%) (%) (%) 5 83.12 11.14 (a) 1.173 (a) 2.43 10 82.57 11.59 (b) 2.226 (ab) 2.34 15 82.56 11.53 (b) 1.955 (ab) 2.25 20 82.69 11.59 (b) 2.882 (b) 2.17 25 81.48 12.53 (d) 3.201 (b) 2.37 30 82.16 12.09 (c) 3.020 (b) 2.34 Pooled SEM 0.66 0.04 0.223 0.11 The different superscripts of the same column values are significantly different (P < 0.05). CBH, carbohydrate. TABLE 4. The soluble protein in the hepatopancreas, glycogen in the hepatopancreas and muscle, and malate dehydrogenase (MDH), pyruvate kinase (PK), and glucose-6-phosphate dehydrogenase (G6PDH) activities of Litopenaeus vannamei fed experimental diets for 42 days. CBH Soluble protein Glycogen in Glycogen in level (%) in hepatopanereas hepatopanereas muscle (mg/g) (mg/mL) (mg/g) 5 113.63 (a) 3.11 (a) 5.02 (abc) 10 I54.97 (b) 4.5 (ab) 5.02 (abc) 15 166.8 (bc) 6.98 (c) 6.93 (c) 20 189.97 (c) 5.33 (bc) 5.53 (bc) 25 166.63 (bc) 4.73 (ab) 4.31 (ab) 30 169.8 (bc) 3.73 (ab) 3.1 (a) Pooled SEM 9.7 0.59 0.62 CBH MDH activities PK activity G6PDH activity level (%) (U/mg protein) (U/mg protein) (U/mg protein) 5 1.71 2.10 0.27 (a) 10 1.12 2.17 0.32 (a) 15 1.72 2.16 0.61 (b) 20 1.61 2.10 0.25 (a) 25 2.46 1.97 0.44 (b) 30 1.18 2.30 0.34 (a) Pooled SEM 0.47 0.18 0.07 The different superscripts of the same column values are significantly different (P < 0.05). CBH, carbohydrate.
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|Author:||Wang, Xiaodan; Li, Erchao; Qin, Jian G.; Wang, Shifeng; Chen, Xuefen; Cai, Yan; Chen, Ke; Hou, Yingm|
|Publication:||Journal of Shellfish Research|
|Date:||Aug 1, 2014|
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