Effects of Lactobacillus pentosus on the growth performance,digestive enzyme and disease resistance of white shrimp,Litopenaeus vannamei (Boone, 1931)

The objective of this study was to evaluate the probiotic properties of lactic acid bacteria (LAB) strains isolated from digestive tract of white shrimp Litopenaeus vannamei. Eighteen LAB colonies were isolated and one bacterium was found capable of producing three extracellular enzymes (protease, cellulose and lipase) simultaneously and exhibited antagonistic activity against shrimp pathogens (Vibrio vulnificus, V. rotiferianus and V. campbellii). The putative probiotic strain AS13 was identified as Lactobacillus pentosus based on 16S rRNA sequencing. The L. vannamei were fed diet containing 0 (control), 10⁶, 10⁷ and 10⁸ CFU g^?1 bacterial cells of AS13 for 28 days. The results showed that supplementation of L. pentosus significantly improved the growth performance and feed utilization in the treated groups over the control. Similarly, digestive enzyme activities were elevated in the intestines of treated groups. Moreover, feeding of supplemented diets containing AS13 significantly reduced the mortality rate caused by pathogenic Vibrio species (V. vulnificus, V. rotiferianus and V. campbellii). Our results indicated L. pentosus AS13 addition at 10⁷ CFU g^?1 can effectively enhance the growth performance, feed utilization, digestive enzymes and disease resistance of L. vannamei in the laboratory condition.     

健康和患病凡纳滨对虾幼虾消化道菌群结构的比较

凡纳滨对虾养殖过程中,早期阶段是病害易感阶段,而消化道菌群结构与对虾健康关系密切。因此,探讨幼虾的消化道菌群尤其是弧菌类细菌与对虾发病的关系对病害有效防治具有重要意义。本实验采用Illumina测序研究了凡纳滨对虾健康和患病幼虾的消化道细菌群落结构,并基于纯培养和16S r DNA,rec A和pyr H基因序列比对,分析了幼虾消化道中弧菌的主要种类组成。结果发现,健康幼虾消化道中α-变形菌纲和厚壁菌门丰度较高,而患病幼虾中γ-变形菌纲、拟杆菌门、放线菌门和β-变形菌纲较高,其中放线菌门丰度差异显著。基于科水平的响应比分析,发现患病幼虾消化道中动性球菌科和噬菌弧菌科的丰度显著降低,而弧菌科的丰度显著升高。相似度分析发现,驱动群落变异的OTU主要来源于弧菌属、海洋杆状菌属、冷杆菌属、假交替单胞菌属以及未分类至属的红杆菌科和微杆菌科。健康幼虾消化道弧菌组成以锡那罗亚州弧菌为主,而患病幼虾消化道弧菌组成以坎氏弧菌为主。尽管健康和患病幼虾消化道内细菌群落结构整体差异不显著,但一些重要类群丰度变化显著,其特征与已知的生态功能一致。     

Impact of a reduced water salinity on the composition of Vibrio spp. in recirculating aquaculture systems for Pacific white shrimp (Litopenaeus vannamei) and its possible risks for shrimp health and food safety

Tropical shrimp, like Litopenaeus vannamei, in land‐based recirculating aquaculture systems (RAS) are often kept at low water salinities to reduce costs for artificial sea salt and the amount of salty wastewater. Although these shrimp are tolerant against low salinities, innate immunity suppression and changes in the microbial composition in the water can occur. As especially Vibrio spp. are relevant for shrimp health, alterations in the species composition of the Vibrio community were analysed in water from six RAS, run at 15‰ or 30‰. Additionally, pathogenicity factors including pirA/B, VPI, toxR, toxS, vhh, vfh, tdh, trh, flagellin genes and T6SS1/2 of V. parahaemolyticus were analysed. The Vibrio composition differed significantly depending on water salinity. In RAS at 15‰, higher numbers of the potentially pathogenic species V. parahaemolyticus, V. owensii and V. campbellii were detected, and especially in V. parahaemolyticus, various pathogenicity factors were present. A reduced salinity may therefore pose a higher risk of disease outbreaks in shrimp RAS. Because some of the detected pathogenicity factors are relevant for human health, this might also affect food safety. In order to produce healthy shrimp as a safe food for human consumption, maintaining high water salinities seems to be recommendable.     

Ingestion of food pellets containing Escherichia coli overexpressing the heat‐shock protein DnaK protects Penaeus vannamei (Boone) against Vibrio harveyi (Baumann) infection

Feeding aquatic animals with bacterial encapsulated heat‐shock proteins (Hsps) is potentially a new method to combat vibriosis, an important disease affecting aquatic animals used in aquaculture. Food pellets comprised of shrimp and containing Escherichia coli overexpressing either DnaK‐DnaJ‐GrpE, the prokaryotic equivalents of Hsp70‐Hsp40‐Hsp20, or only DnaK were fed to juveniles of the white leg shrimp Penaeus vannamei, and protection against pathogenic Vibrio harveyi was determined. Maintaining pellets at different temperatures for varying lengths of time reduced the number of live adhering E. coli, as did contact with sea water, demonstrating that storage and immersion adversely affected bacterial survival and attachment to pellets. Feeding P. vannamei with E. coli did not compromise their survival, indicating that the bacteria were not pathogenic to shrimp. Feeding P. vannamei with pellets containing bacteria overproducing DnaK (approximately 60 cells g^?1 pellets) boosted P. vannamei survival twofold against V. harveyi, suggesting that DnaK plays a role in Vibrio tolerance. Pellets containing DnaK were effective in providing protection to P. vannamei for up to 2 weeks before loss of viability and that DnaK encapsulated by these bacteria enhanced shrimp resistance against Vibrio infection.     

Vibrio spp. Control at Brine Shrimp,Artemia, Hatching and Enrichment

Experiments were conducted to evaluate different prophylactic methods to control the bacterial load in brine shrimp, Artemia, hatching. The first experiment evaluated three treatments to control Vibrio spp. during the Artemia hatching: microalgae (Chaetoceros calcitrans), probiotic (Bacillus spp.), and antibiotic (Florfenicol). In the second experiment, Artemia metanauplius were enriched in distinct treatments with C. calcitrans, probiotic, and emulsion rich in docosahexaenoic and eicosapentaenoic fatty acids. Enriched Artemia metanauplius and nauplii (control) were offered to white shrimp, Litopenaeus vannamei, postlarvae (PL₇–PL₁₉). Presumptive Vibrio were quantified in Artemia, PL, and rearing water. Microalgae and probiotic were effective to control Vibrio spp. in Artemia nauplii. The enrichment process increased the Artemia bacterial load but did not affect Vibrio load in L. vannamei.     

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata in integrated culturing method under zero water exchange system

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata were measured using integrated culturing method under zero‐water exchange system in a 45‐day period. A 2 × 3 factorial design was used with two levels of shrimp stocking densities and three levels of seaweed weight densities. G. corticata was cultured on a net tied to a round polyethylene frame. Culture tanks were filled with 750‐L filtered seawater. A 40‐W compact fluorescent lamp was hung over each tank to provide adequate and sufficient light for seaweed growth. Growth parameters of shrimp and seaweed such as specific growth rate (SGR), weight gained (WG) and average daily growth (ADG) were computed based on the initial and final weight of shrimp and seaweed. The maximum and minimum SGR of L. vannamei (1.97 and 1.69%/day) were observed in treatment S₁A₃ (25 shrimp/m² and 400 g seaweed/m²) and S₂A₁ (50 shrimp/m² without seaweed) respectively. The best survival rate (94.67 ± 1.33%), WG (129.9 ± 2.9%) and feed conversion ratio (1.67 ± 0.04) were also observed in treatment S₁A₃. The SGR of G.corticata in the treatment S₁A₃ (1.97 ± 0.00%/day) was significantly higher, compared to others. Strong positive correlations were obtained between the density of G. corticata and the growth parameters of L. vannamei. The red seaweed G. corticata could boost the growth parameters, survival rate and total production of L. vannamei under zero‐water exchange system.     

Growth performance,haematological parameters,antioxidant status and salinity stress tolerance of juvenile Pacific white shrimp (Litopenaeus vannamei) fed different levels of dietary myo‐inositol

A 10‐week growth trial was run to evaluate effects of myo‐inositol (MI) on growth performance, haematological parameters, antioxidative capacity and salinity stress tolerance of Litopenaeus vannamei. Six practical diets supplemented with graded levels of MI (designated as MI0, MI600, MI1200, MI2400, MI 3600 and MI4800 for 448.8, 974.2, 1568.0, 2810.6, 3835.5 and 4893.6 mg/kg diet, respectively) were fed to six replicate groups of L. vannamei (mean initial body weight 0.63 ± 0.00 g). The results showed that significant increment of growth performance was observed in shrimp fed MI600 diet than those fed MI1200 diet. Lipid concentration in whole body of the shrimp fed MI600 diet was significantly increased. Shrimp fed MI0 diet had lower total protein (TP) as compared to shrimp fed the MI‐supplemented diets (except MI4800 diet). In general, lower activities of antioxidant enzymes and higher malondialdehyde (MDA) content in haemolymph and hepatopancreas were recorded in shrimp fed MI0 diet, compared to those fed the MI‐supplemented diets. Reduced survival after 7‐h salinity stress was present in shrimp fed MI0 diet as compared to those fed MI4800 diet. Dietary MI requirement for glutathione peroxidase activity of L. vannamei was 2705 mg/kg diet.     

Toxicity of the antifouling biocide Sea-Nine 211 to marine algae, crustacea, and a polychaete

We evaluated the acute toxicity of the antifouling biocide Sea-Nine 211 to the algae Chaetoceros calcitrans, Dunaliella tertiolecta, Tetraselmis tetrathele, and Skeletonema costatum, the crustacea Tigriopus japonicus and Portunus trituberculatus, and the polychaete Perinereis nuntia. The algae, and especially the diatoms C. calcitrans and S. costatum, were sensitive to Sea-Nine 211 toxicity, with the average acute toxicity values being 0.32, 3.9, 1.6, 0.22, 1.6, 12, and 27 μg/l for C. calcitrans, D. tertiolecta, T. tetrathele, S. costatum, T. japonicus, P. trituberculatus, and P. nuntia, respectively. A sediment toxicity test for Sea-Nine 211 using the polychaete P. nuntia revealed demonstrated that the 14-day median lethal concentration was 110 μg/kg dry-wt sediment and that growth was the most sensitive indicator. The chronic toxicity values of Sea-Nine 211 for the diatoms C. calcitrans and S. costatum were within the range of reported Sea-Nine 211 concentrations in seawater in coastal Japan, and the toxicity values for P. nuntia were within the reported concentrations in sediment. Based on these results, Sea-Nine 211 may have toxic effects on some sensitive species residing in the coastal areas of Japan, but the ecological risk posed by Sea-Nine 211 would appear to be confined to a limited area of Japanese coastal waters.     

Characteristics and pathogenicity of a Vibrio campbellii-like bacterium affecting hatchery-reared Penaeus indicus (Milne Edwards, 1837) larvae

In the summer months, epizootics occurred regularly among larvae of Penaeus indicus (Milne Edwards) in a hatchery located at Narakkal near Cochin, India. The clinical signs of infected larvae were expansion of chromatophores. opaqueness of body with twisting and degeneration of appendages. Bacteria were isolated from the infected larvae. These bacteria had morphological, physiological and biochemical characteristics of the genus Vibrio, but these properties did not correspond with those of known Vibrio species. The Vibrio was most related to Vibrio campbellii according to DNA-hybridization experiments. By pathogenicity experiments, the present Vibrio isolate was proved to be pathogenic to larvae, postlarvae and adult shrimp. Shrimp larvae challenged by immersion showed significant mortalities. The bacterial isolate failed to produce the infection in adult shrimp after oral inoculation but it caused significant mortality in adults after intramuscular injection. LC⁵⁰ and LD⁵⁰ values of the Vibrio isolate were determined for larvae and adult shrimps, respectively, at different time intervals.     

Effects of dietary marine microalgae,Tetraselmis suecica,on production,gene expression,protein markers and bacterial count of Pacific white shrimp Litopenaeus vannamei

The marine flagellated Chlorophyta Tetraselmis suecica is among the most important live food species in marine aquaculture. In the present study, the effects of dietary supplementation of dried marine microalgae, Tetraselmis suecica, on growth performance; feed utilization; chemical composition; gene expression of superoxide dismutase (SOD), glutathione peroxidase (GPx) and insulin‐like growth factor 2 (IGF‐II) gene of Pacific white shrimp, Litopenaeus vannamei; muscle protein polymorphism; and microbial count were assessed and evaluated. Three hundred and sixty L. vannamei (postlarvae) Pls (0.124 ± 0.002 g) were randomly stocked into 40‐L glass aquaria (30 shrimp/aquarium) and fed three times daily four tested diets: a basal diet (control), diet incorporated with 2.5 g kg^?1 dried T. suecica (T1), 5 g kg^?1 dried T. suecica (T2) and 7.5 g kg^?1 dried T. suecica (T3) in triplicates, for 90 days. At the end of the trial, the survival rate (SR) of L. vannamei fed diets supplemented with different levels of T. suecica was significantly (p < .05) higher than the control diet. The highest weight gain and specific growth rate and the best feed conversion ratio were recorded on L. vannamei fed a diet supplemented with a 7.5 g/kg dried T. suecica. The highest protein, lipid and ash contents were obtained in L. vannamei fed the diet containing 7.5 g/kg T. suecica, when compared with the remaining tested diets. The gene expression of antioxidant genes SOD and GPx was the lowest in the T3 group in comparison with the control group. Meanwhile, expression level of IGF‐II was higher in the T2 group. The total heterotrophic bacterial count was significantly (p < .05) increased with the cumulative T. suecica level, while no significant (p > .05) differences were found in the total Vibrio count among treatments. Overall, the present results have shown that the diet supplemented with the highest inclusion level of dried T. suecica resulted in improved growth and nutrient utilization.     

Antibacterial activity of immobilized silver nanoparticles on TEPA‐Den‐SiO2 against shrimp pathogen,Vibrio sp. Persian1

In this study, we prepared silver nanoparticles immobilized onto silica sand beads as an antibacterial material against pathogenic luminous Vibrio sp. Persian1. Silica beads were modified with 3‐aminopropyltriethoxysilane (APTES), cyanuric chlorid and tetraethylene pentamine, and silver nanoparticles were generated in various concentrations (0.5, 1, and 2 mM g^?1 of silica beads) of AgNO₃ on the surface using chemical reduction. Ag/TEPA‐Den‐SiO₂ samples were characterized by TEM, FE‐SEM/EDS, FT‐IR and ICP OES and their antibacterial activity assayed by zone of inhibition and test tube tests against pathogenic Vibrio sp. The results of the zone inhibitory test revealed that all the Ag/TEPA‐Den‐SiO₂ samples had an antibacterial effect against luminous Vibrio sp. Persian1. In addition, the tube test results showed 100% killing of bacteria in 2 h contact period. Ag/TEPA‐Den‐SiO₂ samples maintained their antibacterial activity after 14‐day immersion in seawater by slow release of silver ions. These results suggest that Ag/TEPA‐Den‐SiO₂ substrates could be effective antibacterial materials for disinfection of seawater used to culture Penaeid shrimp larvae.     

Physiological, metabolic and haematological responses in white shrimp Litopenaeus vannamei (Boone) juveniles fed diets supplemented with astaxanthin acclimated to low-salinity water

The effects of dietary astaxanthin supplemented at 0, 40, 80 or 150 mg astaxanthin kg⁻¹ on growth, survival, moult frequency, osmoregulatory capacity (OC) and selected metabolic and haematological variables in Litopenaeus vannamei acclimated to low‐salinity water (3 g L⁻¹) were evaluated. Supplemented astaxanthin at 80 mg kg⁻¹ improved growth, survival and moult frequency in shrimp. The lowest OC was also exhibited in shrimp fed with dietary astaxanthin at 80 mg kg⁻¹. Shrimp haemolymph concentrations of glucose, lactate, haemocyanin and total haemocyte count were all significantly enhanced by feeding the diet supplemented with 80 mg astaxanthin kg⁻¹ compared with shrimp fed with the other diets. On the basis of these results, dietary astaxanthin supplementation of 80 mg kg⁻¹ is recommended for juvenile L. vannamei cultured in low‐salinity water.     

Differences in uptake and killing of pathogenic and non‐pathogenic bacteria by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei, (Boone)

Phagocytosis is an important function of both invertebrate and vertebrate blood cells. In this study, the phagocytic activity of haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei, (Boone), against pathogenic and non‐pathogenic particles was investigated in vitro. The haemocytes of penaeid shrimp were firstly separated by centrifugation on a continuous density gradient of iodixanol into four fractions with five subpopulations (sub), of which sub 1 (hyalinocytes) and sub 4 (semi‐granulocytes) have the main function in phagocytosis of both pathogenic and non‐pathogenic bacteria as well as fluorescent polystyrene beads. It was found that these haemocyte subpopulations engulfed virulent Vibrio campbellii and Vibrio harveyi at a higher rate than non‐virulent Escherichia coli and polystyrene beads. When these bacteria were mixed with shrimp haemocyte subpopulations and incubated for 180 min, the percentage of viable intracellular V. campbellii (25.5 ± 6.0%) recovered was significantly higher than the percentage recovered from V. harveyi (13.5 ± 1.1%). No viable intracellular E. coli was observed in this study. In contrast to V. harveyi and E. coli, V. campbellii containing endosomes did not acidify in time. Incubation of haemocyte subpopulations with the most virulent V. campbellii strain resulted in a significant drop in haemocyte viability (41.4 ± 6.3% in sub 1 and 30.2 ± 15.1% in sub 4) after 180 min post‐inoculation in comparison with the less virulent V. harveyi (84.1 ± 5.6% in sub 1 and 83.4 ± 4.1% in sub 4) and non‐virulent E. coli (92.7 ± 2.8% in sub 1 and 92.3 ± 5.6% in sub 4) and polystyrene beads (91.9 ± 1.6% in sub 1 and 84.4 ± 3.4% in sub 4). These findings may be a valuable tool for monitoring shrimp health and immunological studies.     

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.     

Effect of polysaccharides extracts of algae Ulva rigida on growth,antioxidant, immune response and resistance of shrimp,Litopenaeus vannamei against Photobacterium damselae

This study was carried out to investigate the effect of water‐soluble polysaccharides extract of algae Ulva rigida (WPU) as dietary supplement on growth performance, antioxidant enzyme activity, lysozyme and phenoloxidase activity, and resistance of shrimp (Litopenaeus vannamei) subjected to bacterial infection with Photobacterium damselae. Three replicate groups of shrimp (1.0 g) were fed four diets containing four levels, 0 or control, 0.5, 1 and 1.5 g/kg of WPU for 8 weeks over the growth trial. Thereafter, 30 shrimps from each dietary treatment were infected with bacteria P. damselae to evaluate disease resistance of infected shrimp. The results of this study showed that WPU was effective as a growth promoter for L. vannamei. The best growth rate was observed in shrimp fed 1.5 g/kg of WPU diet. Regarding antioxidant defences, the diets supplemented with three levels of WPU stimulated glutathione peroxidase and catalase activates in experimental shrimps. MDA content of L. vannamei‐fed diet containing WPU 1.5 and WPU 1.0 was lower than WPU 0 and WPU 0. 5 diets. Also, lysozyme and phenoloxidase activities of shrimp receiving WPU at 1.0 and 1.5 level were significantly higher than those fed WPU 0 and WPU 0.5 diets. In addition, using WPU extract in all diets decreased mortality in L. vannamei in a dose‐dependent manner after challenge with P. damselae. These results suggest that incorporation of water‐soluble polysaccharides from green algae U. rigida at 1.5 g/kg doses improves growth and antioxidant activity and enhances the immune responses in shrimp L. vannamei.     

Some Limiting Factors in Superintensive Production of Juvenile Pacific White Shrimp,Litopenaeus vannamei,in No‐water‐exchange,Biofloc‐dominated Systems

Superintensive shrimp culture in zero‐exchange, biofloc‐dominated production systems is more biosecure and sustainable than traditional shrimp farming practices. However, successful application of this technology depends upon optimizing dietary formulations, controlling Vibrio outbreaks, and managing accumulative changes in water quality and composition. A 49‐d study investigated the effect of two commercial feeds of differing protein content and an indoor limited‐exchange, biofloc‐dominated culture environment on Litopenaeus vannamei performance and tissue composition, water quality and ionic composition, and Vibrio dynamics. Juveniles (5.3 g) were stocked at 457/m³ into four 40 m³ shallow raceways containing biofloc‐dominated water and fed one of two commercial feeds with differing protein content, 35 or 40%. Shrimp performance, Vibrio populations, and changes in shrimp and culture water composition were monitored. There were no significant differences (P > 0.05) in shrimp performance (survival, weight, growth, specific growth rate, total biomass, yield, feed conversion ratio, and protein efficiency ratio) or proximate composition between feed types. The 40% protein feed resulted in higher culture water nitrate and phosphate concentrations, alkalinity consumption and bicarbonate use, and higher phytoplankton density. The presence of Vibrio, specifically Vibrio parahaemolyticus, reduced shrimp survival. This survival decrease corresponded with increased culture water Vibrio concentrations. Culture water K⁺ and Mg²⁺ increased significantly (P < 0.05), and Sr²⁺, Br^?, and Cl^? decreased significantly (P < 0.05) over time. While Cu²⁺ and Zn²⁺ did increase in shrimp tissue, no heavy metals accumulated to problematic levels in culture water or shrimp tissue. These results demonstrate the importance of monitoring Vibrio populations and ionic composition in limited‐exchange shrimp culture systems.     

Vitamin D3 Requirement in Practical Diet of White Shrimp,Litopenaeus vannamei,at Low Salinity Rearing Conditions

A 10‐week feeding trial was conducted to determine the requirements of vitamin D₃ in juvenile Litopenaeus vannamei at low salinity rearing conditions. The basal practical diet contained 685 IU vitamin D₃ per kg diet, Diets 2–6 contained 1960, 3240, 4220, 5630, and 7550 IU vitamin D₃ per kg diet, respectively. Diets were fed twice daily to triplicate groups of L. vannamei (initial average weight 0.39 ± 0.01 g). Growth performance and alkaline phosphatase activity in hepatosomatic were not affected by the supplementation of vitamin D₃. Hepatosomatic index of shrimp fed diets containing vitamin D₃ at 685, 1960, and 3240 IU/kg was lower than the diet containing vitamin D₃ at 7550 IU. Whole‐body moisture and zinc content were significantly decreased with increasing vitamin D₃. Similarly, higher value was also observed on whole‐body protein, ash content, calcium, and phosphorus and magnesium concentration with increasing vitamin D₃. Results of the current study indicated that vitamin D₃ from ingredients could meet the growth requirement but not for whole‐body mineral deposition. Broken‐line regression analysis based on maximum whole‐body ash indicated that vitamin D₃ requirement was recommended to be 6366 IU/kg for juvenile L. vannamei at low salinity rearing conditions.     

Successful rearing of whiteleg shrimp Litopenaeus vannamei larvae fed a desiccation‐tolerant nematode to replace Artemia

The nematode Panagrolaimus sp. was tested as live feed to replace Artemia nauplii during first larval stages of whiteleg shrimp Litopenaeus vannamei. In Trial 1, shrimp larvae were fed one of four diets from Zoea 2 to Postlarva 1 (PL1): (A) Artemia nauplii, control treatment; (NC) nematodes enriched in docosahexaenoic acid (DHA) provided by the dinoflagellate Crypthecodinium cohnii; (N) non‐enriched nematodes; and (Algae) a mixture of microalgae supplemented in C. cohnii cells. In Trial 2, shrimp were fed (A), (NC) and a different treatment (NS) with nematodes enriched in polyunsaturated fatty acids (PUFAs) provided by the commercial product S.presso^®, until Postlarva 6 (PL6). Mysis 1 larvae fed nematodes of the three dietary treatments were 300 μm longer (3.2 ± 0.3 mm) than control larvae. At PL1, control shrimp were 300 μm longer (4.5 ± 0.3 mm) than those fed DHA‐enriched or PUFAs‐enriched nematodes. No differences were observed in length and survival at PL6 between control larvae and those fed DHA‐enriched nematodes (5.1 ± 0.5 mm; 33.1%–44.4%). Shrimp fed microalgae showed a delay in development at PL1. This work is the first demonstration of Panagrolaimus sp. suitability as a complete substitute for Artemia in rearing shrimp from Zoea 2 to PL6.     

Effects of dietary chitosan on growth,survival and stress tolerance of postlarval shrimp,Litopenaeus vannamei

The effect of chitosan, a polymer of glucosamine obtained by the deacetylation of chitin, on growth, survival and stress tolerance was studied in postlarval Litopenaeus vannamei. An experiment was performed with postlarval shrimp (mean initial wet weight 1.2 mg) fed five isoenergic and isonitrogenous diets containing five supplemented levels of chitosan (0, 0.5, 1, 2 and 4 g kg^?1 diet, respectively). The five compound diets (C0, C0.5, C1, C2 and C4) sustained shrimp growth throughout the experiment. Growth performance (final body weights; weight gain; SGR: specific growth rate) in shrimp fed diet C2 was significantly higher than that in shrimp fed diets C0, C0.5 and C1 (P < 0.05), diet C4 treatment provided intermediate growth result. The survival in shrimp fed diet C1 was significantly higher than that in shrimp fed C0 diet (P < 0.05), other diets treatments gave the intermediate survival results. No significant differences were found in growth and survival between diet C2 and C4 treatments. After 9 days of a stress tolerance test, survival in shrimp fed diets C1, C2 and C4 was significantly higher than that in shrimp fed diets C0 and C0.5. We concluded from this experiment that the incorporation of a moderate dietary chitosan was beneficial to the development of postlarval L. vannamei. Considering the effect of chitosan on both growth and survival of postlarval L. vannamei, second‐degree polynomial regression of SGR and survival indicated optimum supplement of dietary chitosan at 2.67 and 2.13 g kg^?1, respectively, so the level of chitosan supplemented in the diet should be between 2.13 and 2.67 g kg^?1.