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.     

Shrimp (Litopenaeus vannamei) production and stable isotope dynamics in clear‐water recirculating aquaculture systems versus biofloc systems

Closed recirculating aquaculture systems (RAS) offer advantages over traditional culture methods including enhanced biosecurity, the possibility of indoor, inland culture of marine species year‐round and potential marketing opportunities for fresh, never‐frozen seafood. Questions still remain regarding what type of aquaculture system may be best suited for the closed‐system culture of marine shrimp. In this study, shrimp (Litopenaeus vannamei) were grown in clear‐water RAS and in biofloc‐based systems. Comparisons were made between the system types with respect to water quality, shrimp production and stable isotope dynamics used to determine the biofloc contribution to shrimp nutrition. Ammonia and nitrite concentrations were higher, and shrimp survival was lower in the biofloc systems. Although stable isotope levels indicated that biofloc material may have contributed 28% of the carbon and 59% of the nitrogen in shrimp tissues, this did not correspond with improved shrimp production. Overall, the water column microbial communities in biofloc systems may be more difficult to manage than clear‐water RAS which have external filters to control water quality. Biofloc does seem to offer some nutritional contributions, but exactly how to take advantage of that and ensure improved production remains unclear.     

Proteome modifications of Pacific white shrimp (Litopenaeus vannamei) muscle under biofloc system

The objective of the study was to examine the effects of biofloc technology on the muscle proteome of Litopenaeus vannamei. Two biofloc treatments and one control were compared: biofloc‐based tanks under zero‐water exchange fed with 150 g/kg crude protein (BF15), or with 250 g/kg crude protein (BF25) diets, and clear water tanks with 50% of daily water exchange stocked with shrimp fed with similar amount of a 250 g/kg crude protein diet, referred to as control. The shrimp (5.28 ± 0.42 g) were divided into the 300‐L fibreglass tanks (water volume of 200 L) at a density of 35 shrimp per tank and were cultured for 35 days. The biofloc groups displayed better growth and survival compared to the control. The muscle tissue from the control and BF25 groups was subjected to proteomic analysis. Lactate dehydrogenase, enolase, arginine kinase, mitochondrial ATP synthase subunit alpha, mitochondrial ATPase inhibitor factor 1 precursor, serpin 3 and myeloid differentiation factor 88 had an increased abundance in the BF25 group, while myosin heavy chain type 1 and myosin heavy chain type 2 showed a decreased abundance. The results indicate that biofloc technology could alter the expression of proteins involved in structure, metabolism and immune status of cultured shrimp.     

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.     

Partial replacement of fishmeal with biofloc meal in the diet of postlarvae of the Pacific white shrimp Litopenaeus vannamei

This study evaluated the potential for the replacement of fishmeal (FM) by biofloc meal (BM) in the diet of Litopenaeus vannamei postlarvae. Four isonitrogenous (437.42 g kg^?1) and isocaloric (19.84 kJ g^?1) diets were formulated, in which FM was replaced with BM at 0% (T0), 7.5% (T7.5), 15% (T15) and 30% (T30). A commercial diet (CD) was used as reference. The study was conducted over 42 days in 50 L tanks connected to a water recirculation system. All tanks were stocked with three postlarvae (initial weight 0.0023 g) per litre. Shrimp survival, weight gain (WG), final weight (FW), specific growth rate (SGR) and protein efficiency ratio (PER) were measured. Survival rates were high (>91.1%) in all treatments. As the T30 treatment returned a significantly (<0.05) better performance in WG, FW, SGR and PER than all other treatments, partial replacement of FM with BM does not appear to affect productivity. In fact, a regression analysis indicated that a FM to BM replacement level of over 20% may actually improve shrimp growth. In addition to providing a cost‐effective alternative ingredient for L. vannamei postlarvae diets, the production of meal also represents a more sustainable way of disposing of the excess of biofloc produced by BFT systems.     

A comparison between water exchange and settling tank as a method for suspended solids management in intensive biofloc technology systems: effects on shrimp (Litopenaeus vannamei) performance,water quality and water use

Biofloc systems rely on microbial processes in the water column to recycle animal waste products, reducing the need for water exchange. These increases biofloc concentration in the water and some form of removal is needed. An experiment was carried out to evaluate two management practices to control biofloc in Litopenaeus vannamei culture. Six tanks (48 m³) were divided into two treatments: water exchange and solid settler. Shrimp were stocked at 164 shrimp m^?2 and with 0.67 g of weight. After 61 days, shrimp under solid settler treatment demonstrated mean weight of 12.7 ± 0.5 g with survival of 73.8 ± 1.4%, and those under water exchange had a final weight of 10.1 ± 0.2 g and survival rate of 57.8 ± 11.1%. Total suspended solids did not differ between the treatments: 326.8 ± 24.9 mg L^?1 for water exchange and 310.9 ± 25.3 mg L^?1 for solid settlers. Settleable solids and productivity/respiration ratio was higher (P < 0.05) in water exchange treatment, indicating differences in physical and biological characteristics of bioflocs. Solids removal method influenced the water use, in which 1150 ± 249 L of water was necessary to produce one kilogram of shrimp using water exchange strategy, and 631 ± 25 L kg^?1 with the use of settlers. Our results indicate that continuous operation of settlers can reduce variability in solids characteristics and water quality variables such as ammonia. Both strategies are efficient in controlling biofloc concentrations of the water; however, settlers can reduce water use and improve shrimp production.     

Effect of using sodium bicarbonate to adjust the pH to different levels on water quality,the growth and the immune response of shrimp Litopenaeus vannamei reared in zero‐water exchange biofloc‐based culture tanks

The bioflocs technology proved to be a sustainable technique used in zero‐water exchange shrimp culture systems. However, the pH and alkalinity may decrease due to the biofloc formation process and Nitrification. A 48‐day experiment was performed to investigate the effects of different pH (7.1–7.6; 7.6–8.1) conditions on water quality, the growth and the health status of shrimp in biofloc technology (BFT) through using sodium bicarbonate to adjust pH respectively. Two pH treatments and one control were compared: T0 — control, T1 — pH 7.6 — NaHCO₃, T2 — pH 8.1 — NaHCO₃, each treatment consisted of three replicate tanks (90 L water volume) and each replicate stocked with 30 shrimp (equivalent to 333 shrimp m^?3). Significant physical, chemical and biological differences (P < 0.05) were detected among treatments. At the end of the experiment, water quality, the growth and the immune response of shrimp in control were significantly lower (P < 0.05) than the other treatments. Moreover, the T2 treatment had a better performance in these three aspects. The results indicated that it was necessary to adjust the pH and alkalinity in the BFT, and a higher pH as well as alkalinity for shrimp growth and the stability of the BFT were more favourable.     

Isolation and identification of Vibrio campbellii as a bacterial pathogen for luminous vibriosis of Litopenaeus vannamei

Outbreak of luminescent disease was reported from Litopenaeus vannamei shrimp farms in Zhangpu County, Southern China during May–July 2011. The clinical signs included fluorescent, less food consumption and high mortality. Bacteria were isolated from the infected shrimps. The pathogen, a luminescent bacterium named VH1 was identified as Vibrio campbellii based on MLSA analysis (16S rDNA, rpoD and toxR). The haemolysin (hly) gene specific in V. campbellii was detected in strain VH1. Pathogenicity test using immersion infection confirmed that strain VH1 was virulent to L. vannamei postlarvae and juveniles, and the LC₅₀ value was 1.55 × 10⁶ CFU mL^?1 and 1.7 × 10⁶ CFU mL^?1 respectively.     

Comparing clear-water RAS and biofloc systems: Shrimp (Litopenaeus vannamei) production,water quality,and biofloc nutritional contributions estimated using stable isotopes

Indoor shrimp aquaculture systems can be used to produce fresh, never-frozen, quality shrimp near metropolitan seafood markets regardless of season and climate. However, questions still remain regarding what type of production system is best suited to maximize indoor production. In this project, two types of systems were compared: clear-water (CW) RAS and biofloc (BF) systems. Three, 1.36 m³ tanks were assigned to each of the two treatments; CW tanks had external settling chambers, two foam fractionators, and external biofilters, all operated continuously. BF tanks had settling chambers and one foam fractionator which were operated as needed to control solids accumulation. Shrimp weighing 0.42 g were stocked in all tanks at 250 m⁻³ and grown for 55 days. Ammonia and pH levels were significantly (P < 0.05) higher in the CW treatment, while nitrite, nitrate, and turbidity were all significantly higher in the BF treatment, although all parameters remained within acceptable ranges for shrimp growth. Shrimp mean harvest weight was significantly higher, biomass (kg m⁻³) was significantly greater, and FCR was significantly lower in the CW treatment; there were no significant differences in survival between treatments. Isotope levels indicated that shrimp in the BF treatment obtained a portion of the C (18-60%) and N (1-18%) in their tissues from biofloc material; however, this effect did not positively influence production in that treatment. By nearly eliminating solids from the water and using an external biofilter, substantially better water quality was maintained in the CW systems, which may have been a major contributor to the improved shrimp production in that treatment.     

Effect of bioflocs on growth and immune activity of Pacific white shrimp,Litopenaeus vannamei postlarvae

The bioflocs technology (BFT) for shrimp production has been proposed as a sustainable practice capable of reducing environmental impacts and preventing pathogen introduction. The microbial community associated with BFT not only detoxifies nutrients, but also can improve feed utilization and animal growth. Biofloc system contains abundant number of bacteria of which cell wall consists of various components such as bacterial lipopolysaccharide, peptidoglycan and β‐1, 3‐glucans, and is known as stimulating nonspecific immune activity of shrimp. Bioflocs, therefore, are assumed to enhance shrimp immunity because they consume the bioflocs as additional food source. Although there are benefits for having an in situ microbial community in BFT systems, better understanding on these microorganisms, in particular molecular level, is needed. A fourteen‐day culture trial was conducted with postlarvae of Litopenaeus vannamei in the presence and absence of bioflocs. To determine mRNA expression levels of shrimp, we selected six genes (prophenoloxidase1, prophenoloxidase2, prophenoloxidase activation enzyme, serine proteinase1, masquerade‐like proteinase, and ras‐related nuclear protein) which are involved in a series of responses known as the prophenoloxidase (proPO) cascade, one of the major innate immune responses in crustaceans. Significant differences in shrimp survival and final body weights were found between the clear water and in the biofloc treatments. mRNA expression levels were significantly higher in the biofloc treatment than the clear water control. These results suggest that the presence of bioflocs in the culture medium gives positive effect on growth and immune‐related genes expression in L.vannamei postlarvae.     

Dietary niacin levels in practical diets for Litopenaeus vannamei to support maximum growth

This study investigated the effects of dietary niacin on growth performance, feed utilization and non‐specific immune response in juvenile Pacific white shrimp. Six isonitrogenous and isolipidic practical diets were formulated with graded niacin levels of 10.9, 65.8, 121.2, 203.4, 387.5 and 769.3 mg kg^?1 of dry diet, respectively. Results indicated that per cent weight gain (WG), specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER) and protein productive value (PPV) were significantly influenced by the dietary niacin levels. The maximum WG and SGR occurred at 121.2 mg kg^?1 niacin diet. However, survival and proximate composition of whole body were not significantly affected by the dietary niacin levels. Dietary niacin levels had no significant effects on the total protein, glucose, triacylglycerol and cholesterol contents in the haemolymph. The activity of catalase and lysozyme in the haemolymph was significantly affected by dietary niacin levels. Based on a two‐slope regression analysis of SGR against dietary niacin level, the dietary niacin requirement of juvenile Pacific white shrimp was 109.6 mg kg^?1.     

Effect of different thermal regimes on glucose,enzymes involved in glycolysis and HSP70 of Litopenaeus vannamei

The purpose of this study was to investigate variations of glucose content, activities of enzymes involved in glycolysis and HSP70 in Litopenaeus vannamei subjected to one constant temperature (25°C) and four daily cyclical temperature change regimes (25 ± 1°C, 25 ± 2°C, 25 ± 3°C and 25 ± 4°C; max 12 am min 12 pm ). Both the glucose and HSP70 in treatment 25°C had a day/night rhythm city (L14:D10), but it gradually disappeared with the increase in temperature fluctuating amplitude. The PK activities varied more and more acutely with the increasing temperature fluctuating amplitude, especially, that in treatment 25 ± 4°C. HK activities were affected by the flux of glucose and the process of glycolysis, which tended to be stable with the increasing temperature fluctuating amplitude. Besides, the variations of PK activities were very abrupt at 25 ± 4°C, which might be unfavourable to the growth of shrimps. The temperature fluctuations affect metabolic adjustments and change the day/night rhythmicity of some physiological indicators.     

Assessment of the relative contribution of dietary nitrogen from fish meal and biofloc meal to the growth of Pacific white shrimp (Litopenaeus vannamei)

The relative contribution of the dietary nitrogen supplied by fish meal and a biofloc meal to the growth of Pacific white shrimp was evaluated using stable isotope analysis. Biofloculated material was obtained from an experimental tilapia culture system. Five formulated diets were supplied. Two of them consisted in isotopic controls having only fish meal or biofloc meal as protein source. Three mixed diets were formulated with varying proportions of these ingredients on a dietary nitrogen basis (75:25, 50:50 and 25:75). At the end of the trial, survival rates were similar (92–100%) but significant differences in mean final weight were observed and a negative correlation between the inclusion of biofloc meal and weight gain was evidenced. Mean final weight in shrimp fed on diet containing only fish meal was 2.8 g, while mean final weight of animals fed on diet containing 50% biofloc was 1.9 g. Isotopic mixing models indicated that all diets contributed higher proportions of dietary nitrogen from fish meal than from biofloc meal. Dietary nitrogen available in diets containing 25%, 50% and 75% of biofloc meal was incorporated in muscle tissue as 5%, 41% and 64% respectively. Diet supplying 25% of nitrogen from biofloc was the only mixed diet eliciting growth comparable to diet containing only fish meal. Lower growth and nitrogen deposition in shrimp fed on diets containing high proportions of biofloc meal were possibly associated to the use of only two protein sources and a restriction of essential amino acids.     

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.     

Effects of Vibrio harveyi on plasma clotting protein (coagulogen) of white shrimp Litopenaeus vannamei

Blood clotting exhibits various important functions, including the prevention of body fluid loss and invasion of pathogens in shrimp. The effects of pathogenic Vibrio harveyi on plasma of white shrimp (Litopenaeus vannamei) in vitro and in vivo were investigated in this study. The clotting protein (coagulogen) in plasma of white shrimp pre‐incubated with extracellular products (ECP) of V. harveyi was found apparently decreased and fast‐migrated in crossed immunoelectrophoresis (CIE) gels. In addition, the coagulogen had been degraded to many low molecular‐weight protein bands in plasma pre‐incubated with ECP on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) gels. When pre‐challenged with bacterial cells and ECP of V. harveyi, the white shrimp began to die at about 30 and 16 h respectively. Moreover, plasma coagulogen was decreased more obvious in shrimp challenged with ECP than that with bacterial cells as visualized in CIE gels, and total plasma protein in both group of shrimp were all decreased. Haemolymph withdrawn from moribund shrimp pre‐challenged with V. harveyi or its ECP was observed unclottable. However, the addition of clotting factors (transglutaminase and/or Ca²⁺) to these unclottable plasma could apparently promote their re‐clotting ability as jelly‐like solid observed in microtubes. The recovery of clotting ability of plasma from moribund shrimp was due to the reformation of coagulogen (200 kDa) after adding the two clotting factors as shown on CIE and SDS‐PAGE gels. The present results suggest that the infection of V. harveyi in white shrimp may not only degrade coagulogen but also influence the presence of transglutaminase and Ca²⁺ ion.     

Improvement of growth performance,water quality and disease resistance against Vibrio harveyi of postlarval whiteleg shrimp (Litopenaeus vannamei) by administration of mixed microencapsulated Bacillus probiotics

The objective of this study was to investigate the effects of mixed Bacillus on growth, water quality and disease resistance against Vibrio harveyi in whiteleg shrimp (Litopenaeus vannamei). Postlarval shrimp (PL₃₀) were fed with (a) a basal diet (the control), (b) a diet containing mixed freeze‐dried Bacillus probiotics (FB) and (c) addition of mixed microencapsulated Bacillus probiotics (MB) in culture water. Addition of FB and MB probiotics improved (p < .05) growth, feed efficiency, survival and culture water quality (ammonia and nitrite) compared to the control group although there was no difference (p > .05) between the two treated groups. Bacillus numbers in gastrointestinal tracts and culture water of FB‐ and MB‐administrated shrimp were higher (p < .05) than in the control. After a 30‐day culture, shrimp were infected with V. harveyi and monitored for 10 days. A significant reduction (p < .05) in cumulative mortality was observed in FB‐ and MB‐supplemented shrimp (43.24% and 45.05%, respectively), compared to the control (63.06%). This finding demonstrated that administration of microencapsulated probiotics was as effective as freeze‐dried probiotics for improving growth, feed efficiency, survival, Bacillus in gastrointestinal tracts, water quality (ammonia and nitrite) and conferring disease resistance to V. harveyi.     

Bacterial population in intestines of white shrimp,Litopenaeus vannamei fed a synbiotic containing Lactobacillus plantarum and galactooligosaccharide

The aims of this study were to unravel the intestinal microbiota of Litopenaeus vannamei after being fed a diet without (control) or with the synbiotic (SYN) for 60 days using next‐generation sequencing technology to see if changes in the intestinal microbiota were involved in the improved growth performance and health status of the shrimp. Next‐generation sequencing data showed that six phyla, 11 classes, 19 orders, 30 families, 58 genera and 73 species with taxonomic names assigned were detected. The majority of the operational taxonomic units (OTUs) was shared between the SYN and control shrimp and comprised 37 OTUs. However, intestinal biodiversity analyses revealed that SYN‐fed shrimp had a higher species richness, evenness and Shannon–Weaver index than did shrimp fed the control diet, but without reaching statistical significance. Interestingly, shrimp fed the SYN diet exhibited improved colonization of Lactobacillus plantarum and reduced prevalences of Vibrio harveyi and Photobacterium damselae in the intestines. These findings indicate that the SYN was able to modulate the intestinal bacterial community of shrimp and could be used to control vibriosis in shrimp.