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.     

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.     

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.     

Acute toxicity of nitrite on white shrimp Litopenaeus vannamei (Boone) juveniles in low‐salinity water

The nitrite toxicity was estimated in juveniles of L. vannamei. The 24, 48, 72 and 96 h LC₅₀ of nitrite‐N on juveniles were 8.1, 7.9, 6.8 and 5.7 mg L^?1 at 0.6 g L^?1; 14.4, 9.6 8.3 and 7.0 mg L^?1 at 1.0 g L^?1; 19.4, 15.4, 13.4 and 12.4 mg L^?1 at 2.0 g L^?1 of salinity respectively. The tolerance of juveniles to nitrite decreased at 96 h of exposure by 18.6% and 54.0%, when salinity declined from 1.0 to 0.6 g L^?1 and from 2.0 to 0.6 g L^?1 respectively. The safe concentrations at salinities of 0.6, 1.0 and 2.0 g L^?1 were 0.28, 0.35 and 0.62 mg L^?1 nitrite‐N respectively. The relationship between LC₅₀ (mg L^?1), salinity (S) (g L^?1) and exposure time (T) (h) was LC₅₀ = 8.4688 + 5.6764S – 0.0762T for salinities from 0.6 to 2.0 g L^?1 and for exposure times from 24 to 96 h; the relationship between survival (%) and nitrite‐N concentration (C) for salinity of 0.6–2.0 g L^?1, nitrite‐N concentrations of 0–40 mg L^?1 and exposure times from 0 to 96 h was as follows: survival (%) = 0.8442 + 0.1909S – 0.0038T – 0.0277C + 0.0008ST + 0.0001CT–0.0029SC, and the tentative equation for predicting the 96‐h LC₅₀ to salinities from 0.6 to 35 g L^?1 in L. vannamei juveniles (3.9–4.4 g) was 96‐h LC₅₀ = 0.2127 S² + 1.558S + 5.9868. For nitrite toxicity, it is shown that a small change in salinity of waters from 2.0 to 0.6 g L^?1 is more critical for L. vannamei than when wider differences in salinity occur in brackish and marine waters (15–35 g L^?1).     

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.     

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.     

Water quality,Vibrio density and growth of Pacific white shrimp Litopenaeus vannamei (Boone) in an integrated biofloc system with red seaweed Gracilaria birdiae (Greville)

An indoor trial was conducted for 42 days to evaluate water quality, Vibrio density and growth of Litopenaeus vannamei in an integrated biofloc system (IBS) with Gracilaria birdiae. Four treatments were used, each in triplicate: Control (monoculture shrimp); IBS 2.5 (L. vannamei and 2.5 kg wet weight seaweed m^?3); IBS 5.0 (L. vannamei and 5.0 kg wet weight seaweed m^?3) and IBS 7.5 (L. vannamei and 7.5 wet weight seaweed m^?3). Shrimp individuals (0.34 ± 0.01 g) were stocked at a density of 500 shrimp m^?3. No water exchange was carried out during the experimental period. Molasses was added once a day as an organic carbon source to maintain the C:N ratio at 12:1. The IBS significantly decreased (P < 0.05) dissolved inorganic nitrogen (DIN) ranging from 19% to 34% (3.12–3.83 mg L^?1), NO₃‐N ranging from 19% to 38% (2.40–3.16 mg L^?1), Vibrio density ranging from by 8–83% (0.40–2.20 log 10³ colony‐forming units mL^?1), and FCR ranging from by 20–30% (1.20–1.37), as compared to the control (4.73 mg L^?1, 3.93 mg L^?1, 2.40 log 10³ colony‐forming units mL^?1, and 1.74 respectively). Moreover, the IBS significantly increased (P < 0.05) crude protein in whole body shrimp, ranging from 8% to 13% (13.2–13.7% wet weight basis); as well as final weight, ranging from 25% to 32% (3.90–4.12 g), weekly growth ranging from 25% to 34% (0.59–0.63 g), and shrimp yield by 22–39% (1.72–1.96 kg m^?3), as compared to control (12.1% wet weight basis, 3.12 g, 0.47 g, and 1.41 kg m^?3 respectively). It can thus be concluded that cultivating Gracilaria birdiae in an IBS with L. vannamei can contribute to DIN and NO₃‐N removal, lower Vibrio density, increased crude protein in whole body shrimp, higher growth and yield parameters in shrimp culture.     

Effects of hypoxia on lysozyme activity and antioxidant defences in the kidney and spleen of Carassius auratus

Assessing the responses of lysozyme activity and antioxidant defences to hypoxia is important to understand the adaptation and tolerance strategies of fish under hypoxia. This study investigated the effects of different dissolved oxygen (DO) levels on Carassius auratus, a natural triploid fish, from Qihe River. The content of malondialdehyde (MDA) and the activities of lysozyme and antioxidant enzymes were measured in the kidney and spleen after hypoxic exposure. At the DO concentrations of 1 and 2 mg L^?1, the activities of antioxidant enzymes and lysozyme significantly decreased and the content of MDA significantly increased (P < 0.01). This result suggests that hypoxia decreased antioxidant enzyme and lysozyme activities and caused MDA accumulation in a concentration‐dependent manner. The DO level of 4 mg L^?1 increased the activities of superoxide dismutase and catalase in the kidney and the activities of glutathione peroxidase and catalase in the spleen (P < 0.05). This result implies that slight hypoxic stress can enhance antioxidant defences to alleviate the damage of oxidative stress. The reduced activities of antioxidant enzymes and the accumulation of MDA at the DO level of ≤2 mg L^?1 implied the decrease in antioxidative ability and the occurrence of oxidative stress. The decrement in lysozyme activity indicated that the antibacterial ability was weakened to some degree. Therefore, hypoxic stress at DO levels ≤2 mg L^?1 should be removed by aeration to avoid the oxidative damage resulting from the reduced antioxidative ability and prevent the outbreak of diseases caused by weakened antibacterial effects.     

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.     

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.     

The efficacy of MS‐222 as anaesthetic agent in four freshwater aquarium fish species

The efficacy of anaesthetic tricaine methanesulfonate (MS‐222) was evaluated in four freshwater aquarium fish species, Zebrafish (Danio rerio), Guppy (Poecilia reticulata), Discu (Symphysodon discus) and Green swordtail (Xiphophorus helleri). The correct dose of anaesthetic should induce the plane 4 of anaesthesia in less than 180 s, recovery in less than 300 s and must survive when exposed during 30 min to anaesthetic. Fishes were exposed to six concentrations of anaesthetic (75, 100, 125, 150, 200 and 250 mg L^?1) and the time of fish reaching plane 4 of anaesthesia, post exposure recovery, and the percentage of survival when fish were subject to 30 min in the anaesthetic were recorded. The optimal doses varied according to the species: D. rerio – 75, 100 and 125 mg L^?1, P. reticulata – 125, 150 and 200 mg L^?1, S. discus – 75 and 100 mg L^?1 and X. helleri – 125 and 150 mg L^?1. The induction time generally decreased significantly with increasing concentration of MS‐222 for all of the species evaluated. The recovery time had a tendency to increase with the increase of the MS‐222 concentration for D. rerio, P. reticulata and S. discus. On the other hand, X. helleri recovery time decreased with the increase of MS‐222 concentration. MS‐222 proved to be effective in anaesthesia for all the freshwater ornamental species studied. The main results clearly show that the optimal dose to anesthetize is fish species dependent and it is completely wrong to extrapolate optimal anaesthetic concentrations between different species.     

Preliminary success using hydrogen peroxide to treat Atlantic salmon,Salmo salar L., affected with experimentally induced amoebic gill disease (AGD)

Currently, the only effective and commercially used treatment for amoebic gill disease (AGD) in farmed Tasmanian Atlantic salmon is freshwater bathing. Hydrogen peroxide (H₂O₂), commonly used throughout the aquaculture industry for a range of topical skin and gill infections, was trialled in vitro and in vivo to ascertain its potential as an alternative treatment against AGD. Under in vitro conditions, trophozoites of Neoparamoeba perurans were exposed to three concentrations of H₂O₂ in sea water (500, 1000 and 1500 mg L^?1) over four durations (10, 20, 30 and 60 min) each at two temperatures (12 and 18 °C). Trophozoite viability was assessed immediately post‐exposure and after 24 h. A concentration/duration combination of 1000 mg L^?1 for >10 min demonstrated potent amoebicidal activity. Subsequently, Atlantic salmon mildly affected with experimentally induced AGD were treated with H₂O₂ at 12 and 18 °C for 15 min at 1250 mg L^?1 and their re‐infection rate was compared to freshwater‐treated fish over 21 days. Significant differences in the percentage of filaments affected with hyperplastic lesions (in association with amoebae) and plasma osmolality were noted between treatment groups immediately post‐bath. However, the results were largely equivocal in terms of disease resolution over a 3‐week period following treatment. These data suggest that H₂O₂ treatment in sea water successfully ameliorated a clinically light case of AGD under laboratory conditions.     

Effects of dietary α‐ketoglutarate supplementation on the antioxidant defense system and HSP 70 and HSP 90 gene expression of hybrid sturgeon Acipenser schrenckii ♀ × A. baerii ♂ exposed to ammonia‐N stress

This study was conducted to determine the effects of dietary α‐ketoglutarate (AKG) supplementation on the antioxidant defense system and gene expression of heat shock protein (HSP) 70 and HSP 90 in hybrid sturgeons Acipenser schrenckii ♀ × A. baerii ♂ exposed to ammonia‐N stress. A 2 × 3 factorial experiment was arranged, in which each diet (0%, 1% AKG) was randomly assigned to 0.25 (control) 5 and 10 mg L^?1 ammonia‐N groups with three replicate aquaria for each 72 h. The 10 mg L^?1 ammonia‐N significantly increased serum ammonia concentrations and intestinal Gln concentrations and GS activity compared with the 0.25 or 5 mg L^?1 ammonia‐N groups. The intestinal Gln concentration and GS activity increased, and the serum ammonia concentration decreased, in fish given dietary supplementation of 1.0% AKG compared with fish given diets without AKG. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity in serum, gills and intestines decreased when fish were exposed to 5 or 10 mg L^?1 ammonia‐N, and their activity increased in fish given diets with 1% AKG. Catalase in the serum and gills decreased when fish were exposed to 5 or 10 mg L^?1 ammonia‐N and increased in fish given diets with 1% AKG. The 10 mg L^?1 ammonia‐N or 1% AKG supplementation increased HSP 70 and HSP 90 gene expression in the liver. The increased activity of antioxidant enzymes, and increased HSP 70 and HSP 90 gene expression in fish fed diets containing 1% AKG suggested higher tolerance to ammonia‐N stress.     

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.     

Performance of a photo‐heterotrophic,hypersaline system for intensive cultivation of white leg shrimp (Litopenaeus vannamei) with minimal water replacement in lined ponds using a stochastic approach

The aim of the study was to evaluate the performance of Litopenaeus vannamei in an intensive photo‐heterotrophic hypersaline system with minimal seawater replacement, and establish relationships between parameters of a stochastic production model and relevant water quality variables. Six experimental 1000 m² lined ponds were stocked at a density of 120 shrimp m^?2 for a 105‐day trial. Salinity increased from 37 to 45 ± 2 g/L, and the water level was maintained with the weekly addition of filtered seawater, equivalent to 1.6% per day. The stochastic model predicted that, at harvest, there is 95% confidence that the system produces between 12.1 and 14.7 t/ha with a mean final individual weight of 13.1 g and a mean survival of 84.2%. Sensitivity analyses showed that dissolved oxygen and individual final weight of shrimp were the main variables influencing yield variance. Nitrogenous compounds were maintained between optimal cultivation levels (NH₃–NH₄⁺ = 0.73 ± 0.43 mg/L, N–NO₂^? = 0.09 ± 0.05 mg/L, N–NO₃^? = 3.22 ± 0.11 mg/L). Heterotrophic bacteria (6.6 ± 3.4 × 10⁵ CFU/ml) and chlorophyll‐α concentration (108.5 ± 80.2 μg/L) showed a similar development pattern, indicating a strong relationship between bacteria and microalgae during cultivation. Vibrio spp. concentrations were low (1.24 ± 1.42 × 10³ CFU/ml). It was shown that the photo‐heterotrophic system could be used in hypersaline conditions, typical of semi‐arid regions, to consistently produce between 12.1 and 14.7 t/ha in 15 weeks.     

Dose‐dependent protective effects of dietary selenium on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper

This study was designed to assess the protective effects of dietary selenium (Se) on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper (Cu). A 60‐day feeding trial was conducted in a static water system for abalone (initial weight: 3.17 ± 0.01 g) exposed to 0.02 mg L^?1 of waterborne Cu. The animals were fed one of the three experimental diets with 0.10, 1.31 and 4.20 mg kg^?1 of Se from Na₂SeO₃·5H₂O respectively. Results showed that the abalone fed 1.31 mg kg^?1 of dietary Se had the lowest Cu concentration in shell, muscle, mantle, gill, hepatopancreas and serum. Meanwhile, the significant lowest contents of malondiadehyde and protein carbonyl in hepatopancreas were also found in the treatment with 1.31 mg kg^?1 of dietary Se (P < 0.05). In addition, this treatment had significant higher glutathione content and thioredoxin reductase activity in hepatopancreas (P < 0.05). However, the activity of Se‐dependent glutathione peroxidase (Se‐GPx) was significantly decreased in the treatment with 4.20 mg kg^?1 of dietary Se (P < 0.05). In this treatment, the protein carbonyl content in hepatopancreas was significantly higher than that in the group with 1.31 mg kg^?1 of dietary Se (P < 0.05). In conclusion, in terms of anti‐oxidation and Cu accumulation, the protective effects of dietary Se on abalone against waterborne Cu were dose‐dependent. The 1.31 mg kg^?1 of dietary Se had this effect, but not 4.20 mg kg^?1 of dietary Se. Moreover, the latter increased the oxidative stress in abalone exposed to the waterborne Cu.     

In vivo evaluation of the effects of commercial Bacillus probiotics on survival and development of Litopenaeus vannamei larvae during the early hatchery period

Three in vivo experiments were conducted to measure the effectiveness of commercial Bacillus probiotics on survival and development of Litopenaeus vannamei larvae from nauplii 3–4 (N_3–4) to zoea 3 (Z₃) stages. Experiment I: Nine commercial Bacillus probiotics were individually added to larvae twice at N₆ and Z₁ at levels of 2, 20, and 100 mg L^?1. Only six of nine products at 20 mg L^?1 exhibited higher or significantly higher larval survival (P < 0.05) than the control. Experiment II: Two superior products from the first experiment were administered to larvae with six dose frequencies at 20 mg L^?1. For both products, three doses, once at each of N_3–4, N₆ and Z₁ stages, yielded the best larval survival and development rates (P < 0.05), and these were confirmed by enhanced activities of tryptase and amylase. Experiment III: The isolates of these two products, identified as Bacillus subtilis and Bacillus licheniformis, were delivered to larvae singly at concentrations of 1.0 × 10⁸ and 1.0 × 10⁹ CFU L^?1, or at the same concentrations by mixing the two equally. At 1.0 × 10⁹ CFU L^?1, B. subtilis exerted more beneficial effects on larvae than B. licheniformis or the mixture. Therefore, the optimal dosage and dose frequency of commercial Bacillus products should be evaluated prior to large‐scale application in shrimp hatcheries to avoid futility or even adverse effects, as spore counts and non‐bacterial ingredients are key factors influencing the efficacy of Bacillus probiotics.     

Effect of different biofloc system on water quality,biofloc composition and growth performance in Litopenaeus vannamei (Boone, 1931)

The experiment was conducted with three biofloc treatments and one control in triplicate in 500 L capacity indoor tanks. Biofloc tanks, filled with 350 L of water, were fed with sugarcane molasses (BFT_S), tapioca flour (BFT_T), wheat flour (BFT_W) and clean water as control without biofloc and allowed to stand for 30 days. The postlarvae of Litopenaeus vannamei (Boone, 1931) with an Average body weight of 0.15 ± 0.02 g were stocked at the rate of 130 PL m^?2 and cultured for a period of 60 days fed with pelleted feed at the rate of 1.5% of biomass. The total suspended solids (TSS) level was maintained at around 500 mg L^?1 in BFT tanks. The addition of carbohydrate significantly reduced the total ammonia‐N (TAN), nitrite‐N and nitrate‐N in water and it significantly increased the total heterotrophic bacteria (THB) population in the biofloc treatments. There was a significant difference in the final average body weight (8.49 ± 0.09 g) in the wheat flour treatment (BFT_W) than those treatment and control group of the shrimp. Survival of the shrimps was not affected by the treatments and ranged between 82.02% and 90.3%. The proximate and chemical composition of biofloc and proximate composition of the shrimp was significantly different between the biofloc treatments and control. Tintinids, ciliates, copepods, cyanobacteria and nematodes were identified in all the biofloc treatments, nematodes being the most dominant group of organisms in the biofloc. It could be concluded that the use of wheat flour (BFT_W) effectively enhanced the biofloc production and contributed towards better water quality which resulted in higher production of shrimp.