Comparing biofloc,clear-water,and hybrid nursery systems (Part I): Shrimp (Litopenaeus vannamei) production,water quality,and stable isotope dynamics

Indoor, intensive, nursery-based recirculating aquaculture systems (RAS) can provide high-quality juvenile shrimp for indoor or pond-based production systems in a biosecure manner. However, it is unclear what type of RAS is most appropriate for indoor shrimp nurseries. This study compared three types of RAS nurseries: biofloc (BF), clear-water (CW), and hybrid (HY). Each treatment included four, randomly assigned 160 L (0.35-m²) tanks that were stocked with 3000 post-larvae shrimp m⁻³. The post-larvae (PL10) shrimp had an initial average weight of 7 ± 0.0 mg and were grown for 48 days. The BF tanks included external settling chambers as the only filtration mechanism. The CW tanks had settling chambers, foam fractionators, and external biofilters to fully clarify the water and process nitrogenous waste. Hybrid tanks included settling chambers, and external biofilters to maintain some suspended solids along with external biofiltration. Overall, the CW treatment had significantly higher dissolved oxygen (DO) and pH levels than the BF and HY systems. The HY treatment had significantly higher DO than the BF treatment. Nitrite concentration was significantly higher in the HY treatment than the CW treatment. Turbidity in the BF treatment was significantly higher than the other treatments. On the final sample date, the BF treatment had significantly higher nitrite and nitrate concentrations than the other treatments. Differences between treatments in terms of shrimp survival, mean harvest weight, specific growth rate, and feed conversion ratio were not significant. The final weight of the shrimp at 48 days for the BF, CW, and HY were 670 mg, 640 mg, and 590 mg respectively. A stable isotope mixing model indicated that, in the BF treatment, 13% of the C and 34% of the N in harvested shrimp tissue may have originated from biofloc material, signifying some nutrient recycling. The nitrification process was more effective with the inclusion of an external biofilter. All three system types appear suitable for RAS shrimp nursery production although consideration should be given to water quality consistency and filtration costs.     

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 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.     

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.     

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.     

Microbiological quality of Litopenaeus vannamei culture using conventional and biofloc systems

Shrimp farming is a fast‐expanding activity that has supported the growth in the supply of these crustaceans to consumers around the world. However, the end product is vulnerable to contamination at all stages of the process, including the rearing tanks, where current practices prioritize to raise stocking densities and the minimization of water renewal. It is thus important to evaluate the potential of these systems for the proliferation of undesirable microorganisms, which may render the product unfit for human consumption. In the present study, the presence of coagulase‐positive Staphylococcus, Salmonella spp., Vibrio spp., and total and thermotolerant coliforms was verified in biofloc tank and conventional pond systems used for the rearing of Litopenaeus vannamei in Pernambuco, Brazil, and the results were compared with the legislation regulating the marketing of fresh shrimp. Samples were collected from two biofloc tanks with a density of 375 shrimp m^?2, and two conventional ponds with 12 shrimp m^?2. None of the samples tested positive for either Salmonella spp. or coagulase‐positive Staphylococcus, which is consistent with the legislation. While no standards are defined legally for Vibrio spp. or coliforms, very low concentrations were recorded in both systems, in comparison with other studies. While some variation in bacterial contamination was observed over the rearing process, the end product of both systems was fit for human consumption. The results of the study indicate that, while the water is not renewed in the biofloc system, the development of undesirable microorganisms can be controlled, with no adverse effects for the end product.     

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.     

Format and mode of artificial substrate fixation affect the performance of Litopenaeus vannamei in high‐density rearing systems

We investigated whether the positive impacts of artificial substrates on shrimp performance are altered in any way by their format or mode of fixation in the tanks. To examine this question, substrates were fixed vertically in the water column in three different configurations: SCF treatment (Substrate Completely Fixed), SPF treatment (Substrate Partially Fixed) and SFF (Substrate in Frond Format). Another treatment received no substrate and served as control (WS = Without Substrate). The shrimp were cultured for 38 days in intensive biofloc culture tanks at a stocking density of 1,125 shrimp m^?3. In general, water quality variables were similar among treatments and remained within the appropriate range for shrimp culture. The final biomass was higher (8.5 kg m^?3) and the feed conversion ratio (FCR) lower (1.6) in all tanks with substrates when compared with the WS treatment tanks (final biomass = 6.3 kg m^?3 and FCR = 3.1). However, only shrimp from the SCF and SPF treatments had a higher survival rate (>95.0%) compared to those in WS tanks (75.9%), which was statistically similar to the SFF treatment (88.0%). These results show that substrate format and its mode of fixation in tanks can alter shrimp performance. In well‐aerated intensive tanks, substrates in frond format are constantly pushed to the tank surface, making it difficult for shrimp to adhere to the screens. In such situation, the extra surface provided by the substrates is not always available to the shrimp, a fact that minimizes the positive effects of substrate.     

Effects of different dietary lipid levels and fatty acids profile in the culture of white shrimp Litopenaeus vannamei (Boone) in biofloc technology: water quality,biofloc composition,growth and health

The objective of this study was to evaluate the influence of different dietary lipid and fatty acids on the nutritional value of bioflocs used as a feed, as well as shrimp performance and health. A total of 1800 Litopenaeus vannamei juveniles (2.87 ± 0.01 g) were cultured in biofloc technology, with a density of 200 shrimp m^?2, and fed with three isoproteic experimental diets at different lipid levels (85 g kg^?1, 95 g kg^?1 and 105 g kg^?1); each treatment was performed in triplicate. After 61 days, no significant difference was observed (P > 0.05) among the water quality parameters. For the shrimp performance, significant difference was observed (P = 0.011) among the values of survival, where treatments with lower lipid levels had higher survival (92.5 ± 3.5% and 91.0 ± 2.5%). Although there are significant differences in survival, no significant differences in the total haemocytes count (THC) were observed. For other growth performance, no differences were observed (P > 0.05). A positive correlation (r = 0.75) has been observed between the dietary oleic acid and bioflocs. The bioflocs showed ‘long‐chain’ polyunsaturated fat acids (lcPUFA), especially arachidonic acid. The shrimp showed similar growth and stayed healthy at the end of the experimental period.     

Optimum dietary protein‐to‐energy ratio for juvenile whiteleg shrimp,Litopenaeus vannamei,reared in a biofloc system

An 8‐week feeding trial was conducted to evaluate the optimum dietary protein‐to‐energy (P/E) ratio in juvenile whiteleg shrimp, Litopenaeus vannamei. Six diets were formulated with two protein levels (30% and 35%) and three digestible energy levels (16, 17.5 and 19 kJ/g diet) at each protein level (₃₀P₁₆, ₃₀P_17.5, ₃₀P₁₉, ₃₅P₁₆, ₃₅P_17.5 and ₃₅P₁₉). Fifty shrimp averaging 0.97 ± 0.03g (Mean ± SD) were randomly distributed in biofloc tanks and fed one of the experimental diets. Weight gain (WG), specific growth rate (SGR) and feed efficiency (FE) of shrimp fed the ₃₅P_17.5 diet were significantly higher than those of shrimp fed ₃₀P₁₆, ₃₀P_17.5 and ₃₀P₁₉ diets (p < .05). Results for non‐specific immune responses showed that diet ₃₅P_17.5 caused higher lysozyme activity in shrimp comparing to lower protein diets. Thiobarbituric acid reactive substances (TBARS) of plasma were lower for shrimp fed on diets consisting of higher protein and energy levels than shrimp fed on the ₃₀P₁₉ diet. Analyses of digestive enzyme activities showed higher trypsin activities for shrimp fed on ₃₅P_17.5 and ₃₅P₁₉ diets comparing to ₃₀P₁₆ and ₃₀P_17.5 diets. Also, hepatopancreatic lipase activity of shrimp fed ₃₅P₁₆, ₃₅P_17.5 and ₃₅P₁₉ diets were significantly higher than those of shrimp fed the other diets. In conclusion, based on the results for growth performance, biochemical parameters, immune responses, oxidative stress and enzyme activities, 35% protein and 17.5 kJ/g digestible energy (₃₅P_17.5) could be considered as the optimum protein‐to‐energy ratio in the diet of juvenile whiteleg shrimp reared in a biofloc system.     

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.     

Replacement of fishmeal by vegetable meal mix in the diets of Litopenaeus vannamei reared in low‐salinity biofloc system: effect on digestive enzymatic activity

Biofloc (consortium of diverse microorganisms associated to suspending substrates) was developed from waste of shrimp Litopenaeus vannamei postlarvae culture under low salinity (5 g L^?1) to provide an additional nutritious biomass and reduce fishmeal inclusion in feeds in a 28‐day indoor shrimp nursery trial conducted in 15 experimental containers (250 L stocked at 600 org m^?3). Four experimental diets (isoproteic and isocaloric) containing different percentage of fishmeal: 0%, 10%, 20% and 30% substituted by vegetable meal mix (corn, sorghum and wheat) were formulated and elaborated. A control treatment consisted of a commercial feed. The main water quality parameters were monitored, and no significant differences were found among treatments. The growth and survival were similar among treatments. In general, digestive enzymatic activities showed differences being greater in the biofloc system compared with clear water. It was concluded that low‐salinity shrimp nursery could be successfully developed with minimum inclusion of fishmeal in feeds, without significant effect on production response. The adjustment of C : N ratio allowed the increase of microbial biomass in the bioflocs, which contributed to maintain good water quality, provide live food and enhance digestive enzymatic activity of cultured organisms.     

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.     

Influence of social factors on the nocturnal activity pattern of the noble crayfish,Astacus astacus (Crustacea,Decapoda) in recirculating aquaculture systems

The social environment plays a major role in the control of growth in many crustaceans, but little is known on the mechanisms involved. Two‐ and three‐year‐old noble crayfish, Astacus astacus are strictly nocturnally active. In the present article, we studied social effects on the pattern of nocturnal activity, represented by the time which the animals spent outside shelters. For this purpose, the activity of groups of uniformly large (‘dominant’) and uniformly small (‘subordinate’) individuals were recorded at different densities both separately and in mixed set‐ups. In crowded laboratory cultures, the presence of larger (dominant) conspecifics was found to reduce the time which smaller (subordinate) individuals spent outside shelters in search of food and feeding by about one‐third. It is suggested that this asymmetric inhibitory effect is an element of the functional chain which ultimately results in an extremely wide variation in individual growth rates of communally reared A. astacus. The relevance of the findings to crayfish aquaculture is discussed.     

Nursery performance of Pacific white shrimp (Litopenaeus vannamei Boone, 1931) cultivated in a biofloc system: the effect of adding different carbon sources

Effect of different carbon sources on nursery performance of Pacific white shrimp (Litopenaeus vannamei) cultivated in biofloc system was investigated. Shrimp postlarvae (98.47 ± 8.6 mg) were fed for 32 days in tanks with water volume of 130 L and density of 1 individual L^?1. One control treatment and four biofloc treatments (BFT1, BFT2, BFT3 and BFT4) with adding different carbon sources including molasses, starch, wheat flour and mixture of them, respectively, were considered at equal weight ratios. According to the results, salinity, dissolved oxygen and pH were not significantly different among the biofloc treatments (P > 0.05). Maximum pH (8.27) and maximum dissolved oxygen (6.35 mg L^?1) were recorded in the control. Maximum (0.43 mg L^?1) and minimum (0.09 mg L^?1) ammonia were recorded in the control and BFT2, respectively (P < 0.05). Using simple carbohydrates (molasses and starch) lowered the ammonia concentration significantly. The highest increase in body weight (1640.43 ± 231.28 mg), growth rate, specific growth rate (8.97 ± 0.42% per day) and biomass (190.29 ± 26.83 mg) were found in BFT1 and the highest survival (90 ± 0.77%) was found in BFT4. The highest feed conversion (1.52 ± 0.23) and the lowest feed efficiency (66.81 ± 7.95) were observed in the control (P < 0.05). The proximate composition analysis revealed an increase in lipid and ash in biofloc treatments. Results indicated that using biofloc technology with zero‐water exchange system and adding carbon sources could help to recycle waste and improve the water quality. Moreover, the type of carbonaceous organic matter as a substrate for heterotrophic bacteria would be effective in degradation and metabolization of ammonia and nitrite.     

Dietary protein level and C/N ratio manipulation in zero‐exchange culture of Litopenaeus vannamei: Evaluation of inorganic nitrogen control,biofloc composition and shrimp performance

A 30‐day experiment was conducted to evaluate inorganic nitrogen control, biofloc composition and shrimp performance in zero‐exchange culture tanks for juvenile L. vannamei offered a 35% (P35) or 25% (P25) crude protein feed, each feed supplemented with additional carbohydrate to increase the C/N ratio to 20:1 (CN20) or 15:1 (CN15). Sucrose was used as a carbohydrate to manipulate the two C/N ratios based on the carbon and nitrogen content of both the feeds and sucrose. The four treatments were referred to as: P35 + CN20, P35 + CN15, P25 + CN20 and P25 + CN15. Each treatment consisted of four replicate tanks (125 L), each stocked with 28 shrimp (equivalent to 224 shrimp m^?3). Bioflocs formed and developed based on initial inoculation in all four treatments; and monitored water quality parameters were maintained within acceptable ranges for shrimp culture throughout the experiment. No significant effects (P > 0.05) of dietary protein level, C/N ratio or their interaction were observed on biofloc development (BFV, TSS and BFVI) and inorganic nitrogen (TAN, NO₂^?‐N and NO₃^?‐N) concentrations. At the end of the experiment, proximate analysis of the bioflocs collected from the four treatments showed crude protein levels of 21.3% ~ 32.1%, crude lipid levels of 1.6% ~ 2.8% and ash levels of 43.4% ~ 61.4%. Extracellular protease and amylase activities of the bioflocs were 9.9 ~ 14.4 U g^?1 TSS and 293.5 ~ 403.8 U g^?1 TSS respectively. Biofloc composition and enzyme activity were both affected by dietary protein level (P < 0.01) and C/N ratio (P < 0.05). Survival, per cent weight gain and protein efficiency ratio of shrimp were not affected (P > 0.05) by dietary protein level, C/N ratio or their interaction; however, the feed conversion ratios were significantly lower (P < 0.05) in treatments with high dietary protein (P35) compared with those in treatments with low dietary protein (P25). The results from this study demonstrate that dietary protein level and C/N ratio manipulation can have important implications for water quality, biofloc composition and shrimp performance in intensive, zero‐exchange biofloc‐based culture systems.     

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).