Ammonia tolerance of Litopenaeus vannamei (Boone) larvae

The tolerance of Litopenaeus vannamei larvae to increasing concentrations of total ammonia nitrogen (TAN) using a short‐term static renewal method at 26°C, 34 g L^?1 salinity and pH 8.5 was assessed. The median lethal concentration (24 h LC₅₀) for TAN in zoea (1‐2‐3), mysis (1‐2‐3) and postlarvae 1 were, respectively, 4.2‐9.9‐16.0; 19.0‐17.3‐17.5 and 13.2 mg L^?1TAN (0.6‐1.5‐2.4; 2.8‐2.5‐2.6 and 1.9 mg L^?1 NH₃‐N). The LC₅₀ values obtained in this study suggest that zoeal and post‐larval stages are more sensitive to 24 h ammonia exposure than the mysis stage of L. vannamei larvae. On the basis of the ammonia toxicity level (24 h LC₅₀) at zoea 1, we recommend that this level does not exceed 0.42 mg L^?1 TAN – equivalent to 0.06 mg L^?1 NH₃‐N – to reduce ammonia toxicity during the rearing of L. vannamei larvae.     

Effects of salinity on standard metabolic rate of juvenile marbled spinefoot (Siganus rivulatus)

Marbled spinefoot, Siganus rivulatus, is a herbivorous euryhaline teleost widely distributed in the Eastern Mediterranean. It is an economically valuable species and a suitable candidate for warm water aquaculture. Accordingly, understanding the effects of environmental factors on fish metabolism is important to optimize culture conditions. Two experiments were performed to establish standard metabolic rate and study the effect of salinity on metabolism of marbled spinefoot. In the first experiment, a series of flow‐through respirometry experiments was performed at 27°C and 35 g L^?1. The standard metabolic rate of marbled spinefoot juveniles was calculated as 0.57 ± 0.02 mg O₂ g^?1 h^?1 (mean ± SE). In the second experiment, fish were maintained at salinities of 25, 30, 35 and 40 g L^?1 for 2 weeks. Flow‐through respirometry was performed to measure respiration rates at the various salinities. Respiration rates were similar among fish in salinities of 30, 35 and 40 g L^?1 but increased significantly at 25 g L^?1. Results suggest that despite the euryhalinity of marbled spinefoot, farmers should maintain salinity within the optimal range of 30–40 g L^?1 in order to improve productivity.     

Acute toxicity of ammonia in Pacu fish (Piaractus mesopotamicus,Holmberg, 1887) at different temperatures levels

Piaractus mesopotamicus juveniles (total length 12 ± 0.5 mm) were exposed to different concentrations of ammonia‐N (un‐ionized plus ionized ammonia as nitrogen), using the static renewal method at different temperature levels (15, 20 and 25°C) at pH 7. The 24, 48, 72, 96 h LC₅₀ values of ammonia‐N in P. mesopotamicus juveniles were 5.32, 4.19, 3.79 and 2.85 mg L^?1 at 15°C; 4.81, 3.97, 3.25 and 2.50 mg L^?1 at 20°C; and 4.16, 3.79, 2.58 and 1.97 mg L^?1 at 25°C respectively. The 24, 48, 72, 96 h LC₅₀ values of NH₃‐N (un‐ionized ammonia as nitrogen) were 0.018, 0.014, 0.013, 0.009 mg L^?1 at 15°C temperature; 0.023, 0.019, 0.016 and 0.012 mg L^?1 at 20°C; 0.029, 0.026, 0.018 and 0.014 mg L^?1 at 25°C. The temperature increase from 15 to 25°C caused an increase of ammonia‐N susceptibility by 21.80%, 9.55%, 31.92% and 30.87%, after 24, 48, 72 and 96 h exposure respectively. Furthermore, we found that exposure of fish to ammonia‐N caused an elevation in total haemoglobin and blood glucose with an increase of 2 mg L^?1 concentration. Ammonia levels tolerated, especially in different temperatures levels, have important implications for the management of aquaculture.     

Larval development and salinity tolerance of Japanese flounder (Paralichthys olivaceus) from hatching to juvenile settlement

Salinity tolerance and growth of Japanese flounder Paralichthys olivaceus at different developmental stages were evaluated, including newly hatched larvae (nhl), yolk sac larvae (ysl), oil droplet larvae (odl), post oil droplet larvae (podl), premetamorphic larvae (preml) and prometamorphic larvae (proml), at 11 salinities from 5 to 55 g L^?1 for 96 h. The ontogenesis during the early life of P. olivaceus was investigated under hatchery salinity 35 g L^?1. The results showed that suitable salinities for nhl, ysl, odl, podl, preml and proml larvae were 10 to 25 g L^?1, 10 to 30 g L^?1, 20 to 30 g L^?1, 30 g L^?1, 10 to 30 g L^?1, 15 g L^?1, respectively, demonstrating an ontogenetic variation of salinity tolerance. The salinity tolerance of nhl, ysl, preml was higher than that of odl, podl and proml. The ysl and preml larvae displayed wide salinity tolerances. The present findings demonstrate that the suitable salinity for larviculture of P. olivaceus is 20–25 g L^?1 before the depletion of oil droplet; after that, higher salinity (30 g L^?1) should be ensured for the post‐oil droplet larvae; the premetamorphic larvae can be cultured at a wide salinity range (10–30 g L^?1), and the metamorphosed larvae should be reared at salinity about 15 g L^?1.     

Acute toxicity of ammonia in Meagre (Argyrosomus regius Asso, 1801) at different temperatures

Argyrosomus regius (3.0 ± 0.9 g) were exposed to different concentrations of ammonia in a series of acute toxicity tests by the static renewal method at three temperature levels (18, 22 and 26°C) at a pH of 8.2. Low temperature clearly increased the tolerance of the fish to total ammonia nitrogen (TAN) and unionized ammonia (NH₃) (P < 0.05). While the 96‐h LC₅₀ values of TAN were 19.79, 10.39 and 5.06 mg L^?1, the 96‐h LC₅₀ of NH₃ were 1.00, 0.70 and 0.44 mg L^?1 at 18, 22 and 26°C respectively. The safe levels of NH₃ for A. regius was estimated to be 0.10, 0.07 and 0.04 mg L^?1 at 18, 22 and 26°C respectively (P < 0.05). This study clearly indicates that A. regius is more sensitive to ammonia than other marine fish species cultured on the Mediterranean and Eastern Atlantic coasts.     

Acute and chronic effects of aqueous ammonia on marbled spinefoot rabbitfish,Siganus rivulatus (Forsskål 1775)

Ammonia is a metabolite of aquatic organisms which might reach deleterious levels in intensive fish farms. The aim of the present study was to determine median lethal concentrations (96‐h LC₅₀) of total ammonia nitrogen (TA‐N) on marbled spinefoot rabbitfish (Siganus rivulatus) and chronic effects of TA‐N on survival, growth and behaviour of juvenile rabbitfish over a 50 day period. In the first experiment, fish were exposed to 0, 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 mg L^?1 TA‐N for 96 h and survival evaluated. In the second experiment, 12 fish were stocked per 50‐L tank and treated with one of 0, 2, 4, 6, 8, 10 and 12 mg L^?1 TA‐N with three replicate tanks per treatment. Survival and growth were determined and histopathological alterations of gills due to chronic ammonia exposure were studied by light and electron microscopy. The 96‐h LC₅₀ values were 16–18 mg L^?1 TA‐N. In the chronic exposure experiment, fish reared in water with 0 mg L^?1 TA‐N had 100% survival and had 50% weight increase in 50 days. Fish at 2 and 4 mg L^?1 TA‐N all died whilst fish in 6, 8, 10 and 12 mg L^?1 TA‐N survived and grew albeit less than in treatment 0 mg L^?1. Gills from ammonia treated fish displayed severe histological and ultrastructural alterations including hyperplasia, hypertrophy and fusion of secondary lamellae, aneurysms and presence of pleomorphic altered cells. Chronic exposure to ammonia is deleterious to marbled spinefoot rabbitfish and low concentrations of ammonia appear to kill the fish in <50 days whilst fish can survive for more than 50 days at concentrations between 6 and 12 mg L^?1 TA‐N.     

Effects of acute change in salinity and moulting on the infection of white leg shrimp (Penaeus vannamei) with white spot syndrome virus upon immersion challenge

In the field, moulting and salinity drop in the water due to excessive rainfall have been mentioned to be risk factors for WSSV outbreaks. Therefore, in this study, the effect of an acute change in environmental salinity and shedding of the old cuticle shell on the susceptibility of Penaeus vannamei to WSSV was evaluated by immersion challenge. For testing the effect of abrupt salinity stress, early premoult shrimp that were acclimated to 35 g L^?1 were subjected to salinities of 50 g L^?1, 35 g L^?1, 20 g L^?1, 10 g L^?1 and 7 g L^?1 or 5 g L^?1 and simultaneously exposed to 10^5.5 SID₅₀ mL^?1 of WSSV for 5 h, after which the salinity was brought back to 35 g L^?1. Shrimp that were transferred from 35 g L^?1 to 50 g L^?1, 35 g L^?1 and 20 g L^?1 did not become infected with WSSV. Shrimp became infected with WSSV after an acute salinity drop from 35 g L^?1 to 10 g L^?1 and lower. The mortality in shrimp, subjected to a salinity change to 10 g L^?1, 7 g L^?1 and 5 g L^?1, was 6.7%, 46.7% and 53.3%, respectively (P < 0.05)_. For testing the effect of moulting, shrimp in early premoult, moulting and post‐moult were immersed in sea water containing 10^5.5 SID₅₀ mL^?1 of WSSV. The resulting mortality due to WSSV infection in shrimp inoculated during early premoult (0%), ecdysis (53.3%) and post‐moult (26.72%) demonstrated that a significant difference exists in susceptibility of shrimp during the short moulting process (P < 0.05). The findings of this study indicate that during a drop in environmental salinity lower than 10 g L^?1 and ecdysis, shrimp are at risk for a WSSV infection. These findings have important implications for WSSV control measures.     

Effect of γ‐aminobutyric acid supplementation on growth performance,endocrine hormone and stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei,fed low fishmeal diet

An 8‐week feeding trial was conducted to evaluate the optimum dietary γ‐aminobutyric acid (GABA) level in low fishmeal diet for juvenile Pacific white shrimp, Litopenaeus vannamei. Six practical diets (449 g kg^?1 protein, 87 g kg^?1 lipid) supplemented with graded levels of GABA (0, 50, 100, 150, 200, 250 mg kg^?1) were formulated. Each diet was randomly assigned to triplicate groups of 30 shrimps (approximately 0.44 g), and the shrimps were fed four times a day to apparent satiation. Weight gain and gain rate were significantly increased with the supplementation of GABA (P < 0.05). Hepatosomatic index and survival were also significantly influenced by the dietary GABA levels (P < 0.05) and show a similar trend to those of growth performance. Insulin and neuropeptide Y concentrations in blood were increased with the supplementation of GABA. In total, 150 mg GABA kg^?1‐supplemented diets significantly increased the survival after 12, 24 and 36 h of NH₃ stress, also increased the total antioxidant capacity, total superoxide dismutase, catalase, antihydroxyl radical and phenoloxidase activities before and after the 36 h NH₃ stress compared to basic group. These results clearly indicated that GABA could improve growth performance, antioxidative capacity and resistance against NH₃ stress in L. vannamei, and 150 mg kg^?1 GABA supplementation was suitable for L. vannamei fed with low fishmeal diet.     

A diet of fructose‐enriched Artemia improves the response of juvenile Litopenaeus vannamei shrimp to acute low‐salinity challenge

The Pacific white shrimp (Litopenaeus vannamei), as an economically important species, has been reared in low‐salinity water during the last decade. To investigate how juvenile L. vannamei shrimp fed with fructose‐enriched Artemia respond to acute low‐salinity stress, the shrimp were randomly divided into four treatment groups, three groups were fed with Artemia enriched with either 100, 200 and 300 mg L^?1 of fructose and a control group fed with Artemia with no enrichment for 10 days. The results showed that the 300 mg L^?1 fructose group demonstrated the maximum survival rate and glycogen content. Additionally, the 300 mg L^?1 fructose group showed significantly higher Na⁺/K⁺‐ATPase activity, total antioxidant capacity, and expression levels of Na⁺/K⁺‐ATPase α‐subunit, V‐H ATPase α‐subunit, hexokinase, phosphofructokinase, antioxidants (SOD, CAT, GPX) and Hsp70 mRNA when compared with the control group. Furthermore, after exposure to low salinity, the mRNA levels of phosphofructokinase, V‐H ATPase α‐subunit, GPX, p38, JNK and Rac1 stayed constant in shrimp fed with fructose‐enriched Artemia but changed significantly in the control group. Thus, a diet of fructose‐enriched Artemia can improve the osmoregulation and survival of juvenile L. vannamei shrimp exposed to low salinities.     

Survival and respiration of marbled rabbitfish (Siganus rivulatus) fingerlings at various oxygen tensions

Marbled rabbitfish, Siganus rivulatus, is an economically valuable herbivorous fish and a potential candidate for warmwater aquaculture. This study was carried out to: (1) assess the effect of various oxygen concentrations on survival and behaviour of S. rivulatus fingerlings and (2) investigate the response of S. rivulatus to hypoxia and determine its critical oxygen tension (P_crit). In the first experiment, groups of rabbitfish (15 fish per group) were maintained for 1 h in waters of various oxygen concentrations. They were then transferred to well‐aerated tanks and observed for 72 h. Survival was recorded, fish behaviour at low oxygen concentrations observed, and LC₅₀ after 1‐h hypoxia and 72‐h recovery evaluated. In the second experiment, a series of stop‐flow respirometry experiments were performed during which dissolved oxygen was allowed to drop to 0.5 mg L^?1 and respiration rate recorded at various oxygen concentrations. In the first experiment, all fish survived for 1 h at oxygen concentration of 1.44 mg L^?1 and greater, but started dying at oxygen concentrations below 0.65 mg L^?1 (16% survival). The LC₅₀ of S. rivulatus fingerlings was 0.6 mg L^?1. Results of the second experiment showed that S. rivulatus is an oxyregulator until P_crit (1.7 mg L^?1 O₂) is reached, becoming an oxyconformer below this concentration. Findings allow for a better understanding of environmental oxygen tolerances and minimum acceptable oxygen concentration in rabbitfish aquaculture.     

Effect of different total suspended solids levels on a Litopenaeus vannamei (Boone, 1931) BFT culture system during biofloc formation

In a Biofloc Technology System (BFT), there is constant biofloc formation and suspended solids accumulation, leading to effects on water quality parameters that may affect the growth performance of cultured shrimp. This study aimed to analyse during biofloc formation the effect of different total suspended solids (TSS) levels on water quality and the growth performance of Litopenaeus vannamei shrimp in a BFT system. A 42‐day trial was conducted with treatments of three ranges of TSS: 100–300 mg L^?1 as low (TL), 300–600 as medium (TM) and 600–1000 as high (TH). The initial concentrations of 100 (TL), 300 (TM) and 600 mg L^?1 (TH) were achieved by fertilization before starting the experiment. Litopenaeus  vannamei juveniles with an average weight of 4.54 ± 1.19 g were stocked at a density of 372 shrimp m^?3. Physical and chemical water parameters and shrimp growth performance were analysed. After 6 weeks, TSS mean concentrations were 306.37, 532.43 and 745.2 mg L^?1 for, respectively, TL, TM and TH treatments. Significant differences (P < 0.05) were observed in TSS, settleable solids, pH, alkalinity and nitrite, especially between the TL and TH treatments. Similarly, differences (P < 0.05) were observed in the growth performance parameters, specifically final weight, survival, feed conversion and productivity. The water quality parameters at lower range of total suspended solids concentration (TL) treatment resulted in a better performance of L. vannamei in the BFT system. The maintenance at range of 100–300 mg L^?1 TSS is thus important to the success of shrimp culture.     

Effects of aqueous Na/K and dietary K on growth and physiological characters of the Pacific white shrimp,Litopenaeus vannamei,reared in low‐salt well water

The synchronous effects of aqueous Na/K and dietary potassium (K⁺) on growth and physiological characters was studied on the Pacific white shrimp (Litopenaeus vannamei) reared in low‐salinity well water (4 ppt) for 8 weeks with initial weight of 0.28 ± 0.01 g. Three practical diets were formulated with supplement of 0, 0.3%, 0.6% K⁺ which contained 1.29 g/100 g, 1.60 g/100 g, 1.93 g/100 g K⁺ respectively. The supplement of K⁺ to the low‐salinity well water was 10, 20, 40 mg L^?1 which formed Na:K ratios of 42:1, 33:1, 23:1 respectively. Results showed that when the aqueous Na:K ratio was 42 and dietary K⁺ was 1.93 g/100 g K⁺, the WGR and PER of L. vannamei were the highest and the FCR was lower than that of others (P < 0.01). Supplement of K⁺ into well water and diets did not showed significant effects on haemolymph ammonia‐N, uric acid, urea content (P > 0.05), but had a extremely significant effect on arginase activity and Cl^? concentration (P < 0.01). Moreover, similar results were observed in alkaline phosphatase (ALP), bacteriolytic activity (LSZ) and respiratory burst activity (O₂^?) (P < 0.05). These results suggested that aqueous Na/K in the low‐salt well water and dietary K had significant synergistic effect on the growth, osmoregulation and immunity of L. vannamei. Concluded from the growth performance, nitrogen metabolism, osmoregulation and immunity, as the Na/K in the low‐salinity well water descended from 42 to 23, the requirement of dietary K⁺ was also decreased.     

Effects of incubation water hardness and salinity on egg hatch and fry survival of Nile tilapia Oreochromis niloticus (Linnaeus)

This study examined the effects of water hardness and salinity on yolk sac larvae and swim‐up fry survival of Nile tilapia, Oreochromis niloticus (Chitralada strain), eggs during artificial incubation. Four experiments were conducted to evaluate the effects of hardness, salinity and the sources of saline incubation water. High water hardness treatments (500–4200 mg L^?1 as CaCO₃) resulted in higher yolk sac larvae and swim‐up fry survival than low water hardness treatments (50.0 and 132 mg L^?1 as CaCO₃); although yolk sac larvae and swim‐up fry survival did not differ among the high or low hardness treatments. Salinity of 4.0 g L^?1 using seawater, and 4.0 and 8.0 g L^?1 using unprocessed common salt resulted in the higher survival rate of yolk sac larvae and swim‐up fry than other salinity treatments. Yolk sac larvae and swim‐up fry survival was found to decrease with the increase in salinity and increase with the increase in water hardness. The present study demonstrated the positive effects of increased water hardness level (>132 mg L^?1) on yolk sac larvae and swim‐up fry survival. The study also showed that seawater salinity of 4 g L^?1 was the most appropriate salinity level for incubating Nile tilapia eggs.     

Effect of salinity on survival, growth, oxygen consumption and ammonia-N excretion of juvenile whiteleg shrimp, Litopenaeus vannamei

In this study, we tested the lower salinity tolerance of juvenile shrimps (Litopenaeus vannamei) at a relatively low temperature (20 °C). In the first of two laboratory experiments, we first abruptly transferred shrimps (6.91 ± 0.05 g wet weight, mean ± SE) from the rearing salinity (35 000 mg L^?1) to salinities of 5000, 15 000, 25 000, 35 000 (control) and 40 000 mg L^?1 at 20 °C. The survival of L. vannamei juvenile was not affected by salinities from 15 000 to 40 000 mg L^?1 during the 96‐h exposure periods. Shrimps exposed to 5000 mg L^?1 were significantly affected by salinity, with a survival of 12.5% after 96 h. The 24‐, 48‐ and 96‐h lethal salinity for 50% (LS₅₀) were 7020, 8510 and 9540 mg L^?1 respectively. In the second experiment, shrimps (5.47 ± 0.09 g wet weight, mean ± SE) were acclimatized to the different salinity levels (5000, 15 000, 25 000, 35 000 and 40 000 mg L^?1) and then maintained for 30 days at 20 °C. Results showed that the survival was significantly lower at 5000 mg L^?1 than at other salinity levels, but the final wet weight under 5000 mg L^?1 treatment was significantly higher than those under other treatments (P<0.05). Feed intake (FI) of shrimp under 5000 mg L^?1 was significantly lower than those of shrimp under 150 00–40 000 mg L^?1; food conversion efficiency (FCE), however, showed a contrasting change (P<0.05). Furthermore, salinity significantly influenced the oxygen consumption rates, ammonia‐N excretion rates and the O/N ratio of test shrimps (P<0.05). The results obtained in our work provide evidence that L. vannamei juveniles have limited capacity to tolerate salinities <10 000 mg L^?1 at a relatively low temperature (20 °C). Results also show that L. vannamei juvenile can recover from the abrupt salinity change between 15 000 and 40 000 mg L^?1 within 24 h.     

Menthol as anaesthetic for lambari Astyanax altiparanae (Garutti & Britski 2000): attenuation of stress responses

The anaesthetic potential of menthol was evaluated in lambari Astyanax altiparanae by exposing fingerlings to concentrations 50, 100, 150, 200, 250 and 300 mg L^?1 and measuring the induction and recovery times to deep anaesthesia, the mortality rates during and 96 h after procedure and after 6 min of continuous exposure. The effect of menthol on stress responses were evaluated by comparing glucose and cortisol levels of juveniles subjected to anaesthesia (50 mg L^?1), stress (air exposure) or pre‐anaesthesia associated to stress. All concentrations induced deep anaesthesia within 0.5 to 1 min, with recovery between 1.83 and 4.16 min, without mortality during the induction or up to 96 h after exposure. Induction time decreased and recovery time increased linearly as the menthol concentration increased. Continuous exposure to 50, 100 and 150 mg L^?1 concentrations resulted in mortality rates of 0%, 20% and 80% respectively. Anaesthesia or air exposure increase blood glucose but prior anaesthesia with menthol suppressed the elevation of cortisol caused by stress. Menthol has an anaesthetic effect and attenuates the stress response in lambari and 50 mg L^?1 is the most effective concentration for inducing deep anaesthesia in 1.0 min, safe for up to 6 min exposure.     

Acute tolerance of juvenile Florida pompano, Trachinotus carolinus L., to ammonia and nitrite at various salinities

The acute tolerance of juvenile Florida pompano Trachinotus carolinus L. (mean weight±SE=8.1±0.5 g) to environmental unionized ammonia‐nitrogen (NH₃‐N) and nitrite‐nitrogen (NO₂‐N) at various salinities was determined via a series of static exposure trials. Median‐lethal concentrations (LC₅₀ values) of NH₃‐N and NO₂‐N at 24, 48, and 96 h of exposure were calculated at salinities of 6.3, 12.5 and 25.0 g L^?1 at 28 °C (pH=8.23–8.36). Tolerance of pompano to acute NH₃‐N exposure was not affected by salinity, with 24, 48 and 96 h LC₅₀ values ranging from 1.05 to 1.12, 1.00 to 1.08 and 0.95 to 1.01 mg NH₃‐N L^?1 respectively. Regarding NO₂‐N, tolerance of pompano to this environmental toxicant was compromised at reduced salinities. Median‐lethal concentrations of NO₂‐N to pompano at 24, 48 and 96 h of exposure ranged from 67.4 to 220.1, 56.9 to 140.7 and 16.7 to 34.2 mg NO₂‐N L^?1 respectively. The results of this study indicate that juvenile Florida pompano are relatively sensitive to acute NH₃‐N and NO₂‐N exposure, and in the case of the latter, especially at lower salinities.     

Dietary vitamin C mitigates monocrotophos toxicity in the freshwater fish Clarias batrachus

The amelioration effect of balanced diet with supplementation of vitamin C against stress was evaluated in freshwater paddy field fish, Clarias batrachus, exposed to organophosphate insecticide, monocrotophos (MCP). Three groups of fishes, that is, (E₀) Control, (E₁) MCP (2.14 mg L^?1) exposure and E₂) Vitamin C, MCP (0.2 g kg^?1 body weight + 2.144 mg L^?1) were selected for the sublethal study. The effect of MCP on protein metabolism in physiological sensitive tissues, namely gills, kidney, liver and muscle of the fish has been determined. Stress on Clarias batrachus was evident that the total protein, amino acid and ammonia contents were decreased, and urea and glutamine levels were elevated in all tissues for 30 days. Similarly, the activities of protease, aminotransferases and phosphatases were elevated significantly in all the tissues during the same period. Fish fed with a diet supplemented by vitamin C (0.2 g kg^?1 body weight) could significantly counter these effects.     

Quantitative dietary leucine requirement of juvenile Pacific white shrimp,Litopenaeus vannamei (Boone) reared in low‐salinity water

The experiment was conducted to determine the leucine requirement of juvenile Pacific white shrimp Litopenaeus vannamei (Boone) in low‐salinity water (0.50–1.20 g L^?1). Six diets were formulated to contain 410 g kg^?1 crude protein with fish meal, peanut meal and precoated crystalline amino acids with different concentration of l ‐leucine (16.72, 19.60, 22.06, 24.79, 27.28 and 30.16 g kg^?1 dry diet). Each diet was randomly assigned to triplicate groups of 30 shrimps (0.38 ± 0.002 g), and the feed trial lasted for 8 weeks. The results indicated that the maximum weight gain was observed at 24.95 g kg^?1 dietary leucine group, whereas the diets containing higher leucine concentration conversely reduced the growth performance (P < 0.05). Moreover, the highest body protein content and body protein deposition and the lowest haemolymph AST and ALT activities were also found at 24.95 g kg^?1 dietary leucine group. With the increase in leucine in diets, a dose‐dependent increase was found in body lipid content and haemolymph urea concentration. The polynomial regression calculated using weight gain, feed efficiency and body protein deposition indicated that the optimal dietary leucine requirement for L. vannamei reared in low‐salinity water was 23.73 g kg^?1 leucine of dry diet, correspondingly 57.88 g kg^?1 of dietary protein.     

Acute toxicity of nitrite to mud crab Scylla serrata (Forsskål) larvae

Early larval stages of mud crab Scylla serrata were exposed to different concentrations of nitrite (40, 80 and 160 mg L⁻¹ and a control, without added nitrite) and three salinity levels (25, 30 and 35 g L⁻¹) using a static renewal method. No interactive effect of nitrite and salinity was detected. Estimated LT₅₀ in 96‐h toxicity tests decreased in all stages with increasing nitrite concentrations in all salinity levels. The 96‐h LC₅₀ values of nitrite‐N were 41.58, 63.04, 25.54, 29.98 and 69.93 mg L⁻¹ for zoea 1, 2, 3, 4 and 5 respectively. As the larvae grew, they showed a progressive increase in tolerance to nitrite. The toxicity of nitrite to larvae increased with exposure time. The median lethal concentration was not affected by salinity. The chloride component of salinity within 25–35 g L⁻¹ did not seem to be as effective in alleviating toxicity as has been reported in other crustacean species. Based on 96‐h LC₅₀ and an application factor of 0.1, the ‘safe level’ of rearing mud crab larvae was calculated to be 4.16, 6.30, 2.55, 2.99 and 6.99 mg L⁻¹ nitrite‐N for zoea 1, 2, 3, 4 and 5 respectively.     

Effect of l‐ascorbyl‐2‐polyphosphate supplementation on growth performance,body composition,antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp,Litopenaeus vannamei

An 8‐week feeding trial was conducted to evaluate the effects of ascorbic acid (AsA), in the form of l ‐ascorbyl‐2‐polyphosphate (LAPP) on growth performance, body composition, antioxidative capacity and salinity stress tolerance of juvenile Pacific white shrimp, Litopenaeus vannamei. Five practical diets (46% crude protein and 7.6% lipid) supplemented with graded levels of AsA (14.64, 48.55, 84.98, 308.36 and 639.27 mg kg^?1 diet) were fed to five replicate groups of L. vannamei (mean initial wet weight 0.57 g). No significant differences were found on growth performance among all treatments. However, whole body lipid content significantly decreased with dietary AsA levels increasing. Activities of total antioxidant capacity, glutathione reductase and glutathione peroxidase were significantly affected by dietary AsA levels. Shrimp fed LAPP‐free diet had higher malondialdehyde content than those fed the diets supplemented with LAPP. Dietary AsA levels higher than 308.36 mg kg^?1 diet increased the survival of shrimps after 1, 2 and 3 h of acute salinity change. Broken‐line regression analysis on survival after 3 h of salinity stress and second‐degree polynomial regression analysis on glutathione reductase data indicated that the optimal dietary AsA requirement of L. vannamei was estimated to be 306.39, 319.75 mg kg^?1 diet respectively.