Water quality and growth of Pacific white shrimp Litopenaeus vannamei (Boone) in co-culture with green seaweed Ulva lactuca (Linaeus) in intensive system

An indoor trial was conducted for 28 days to evaluate the effects and interactions of biofloc and seaweed Ulva lactuca in water quality and growth of Pacific white shrimp Litopenaeus vannamei in intensive system. L. vannamei (4.54 ± 0.09 g) were stocked in experimental tanks at a density 132 shrimp m^?2 (566 shrimp m^?3) and the U. lactuca was stocked at a density 0.46 kg m^?2 (2.0 kg m^?3). Biofloc with seaweed (BF-S) significantly reduced (P < 0.05) total ammonia nitrogen (TAN) by 25.9 %, nitrite–nitrogen (NO₂–N) by 72.8 %, phosphate (PO ₄ ³ -P) by 24.6 %, and total suspended solids by 12.9 % in the water and significantly increased (P < 0.05) settleable solids by 34.2 % and final weight of shrimp by 6.9 % as compared to biofloc without seaweed. The BF-S can contribute by reducing nitrogen compounds (TAN and NO₂–N), phosphate (PO ₄ ³ -P), and total suspended solids in water and increased final weight of shrimp.     

Effects of carbohydrate supplementation on water quality,microbial dynamics and growth performance of giant tiger prawn (Penaeus monodon)

Biofloc technology based 45-day indoor growth trial was conducted to evaluate the effect of carbohydrate (CHO), molasses supplementation on two developmental stages viz, juvenile (J) and sub-adult (SA) of Penaeus monodon in zero-water exchange system. P. monodon juveniles (1.56 ± 0.04 g) and sub-adults (14.32 ± 0.22 g) were stocked in fibre-reinforced plastic tanks (1,000 L) with soil base. Carbohydrate supplementation significantly reduced (p < 0.05) the total ammonia nitrogen (TAN), nitrite-N (NO₂-N) and nitrate-N (NO₃-N) over the time periods in sub-adult group. However, no significant difference in TAN and NO₂-N was observed in juvenile-based treatments. Supplementation of carbohydrate significantly increased (p < 0.05) the water and soil total heterotrophic bacterial (THB) count in both juvenile and sub-adults. Despite increase in total Vibrio count (TVC), lower TVC/THB % was noticed in the water column of J + CHO (0.82) and SA + CHO (0.73) compared to control groups, J ? CHO (1.48) and SA ? CHO (1.21). Supplementation of carbohydrate increased the final body weight in J + CHO (6.51 ± 0.44 g) and SA + CHO (22.52 ± 0.98 g) compared with respective controls, J ? CHO (5.05 ± 0.45 g) and SA ? CHO (20.00 ± 0.33 g). Similarly, significantly lower (p < 0.05) feed conversion ratio and better protein efficiency ratio were recorded in carbohydrate-supplemented juvenile and sub-adults treatment groups. The present study demonstrates that supplementation of carbohydrate reduced the nitrogenous metabolites and significantly enhanced the growth performance of juvenile and sub-adult of P. monodon.     

Effect of the C/N ratio on inorganic nitrogen control and the growth and physiological parameters of tilapias fingerlings,Oreochromis niloticu reared in biofloc systems

The objective of this study was to document the effects of different C/N ratios on inorganic nitrogen control and the growth and health of tilapia (Oreochromis niloticus) fingerlings reared in biofloc systems. The fish (0.50 ± 0.02 g) were randomly assigned to the control group (clear water) or to one of three biofloc groups (added glucose) with a C/N ratio of 10, 15, or 20 (CN‐10, CN‐15, or CN‐20, respectively). All four groups were cultured for 120 days. The inorganic nitrogen dynamics showed that heterotrophic and autotrophic bacteria existed in the low C/N ratio group, and heterotrophic bacteria dominated the high C/N ratio group. In addition, growth was significantly better in the CN‐10 and CN‐15 groups (15.67 ± 0.78 g and 14.75 ± 0.54 g, respectively) compared to the CN‐20 (13.04 ± 0.94 g) and control (13.04 ± 0.46 g) groups (p < .05). Activities of the digestive enzymes lipase and trypsin were significantly higher in the CN‐10 and CN‐15 groups than in the control group (p < .05). In addition, activities of lysozyme, alkaline phosphatase, superoxide dismutase, glutathione and malondialdehyde were significantly higher in the CN‐10 and CN‐15 groups than in the control group (p < .05). Finally, the concentrations of glucose and cortisol in the biofloc systems were significantly lower than those in the control group (p < .05). However, the concentrations of triglyceride, aspartate transaminase and alanine aminotransferase did not differ significantly among the four groups (p > .05). At the end of the trial, an acute crowding and hypoxia stress test performed for 3 days showed that the CN‐10 and CN‐15 groups had the best survival.     

Biofloc technology with addition molasses as carbon sources applied to Litopenaeus vannamei juvenile production under the effects of different C/N ratios

The current study was conducted to evaluate the effects of increasing carbon to nitrogen (C/N) ratios on water quality, growth performance, and body composition of Litopenaeus vannamei juveniles. Shrimp with initial average weight of 2.50?±?0.3 g were cultivated for 35 days in 300-L tanks (160 L of water volume) with a density of 1 g per liter. The experiment was performed in five treatments with three replicates. One control group and four biofloc treatments with different C/N ratio were considered: C/N of 10 (CN10), 14 (CN14), 18 (CN18), and 22 (CN22). Shrimp were fed three times a day, and molasses just was added as a carbon sources to the biofloc treatments after each feeding. According to the results, the lowest amount of dissolved oxygen (5.33 mg L^?1) and pH (7.83) was observed in CN22 treatment, which showed a significant difference with other treatments (P?<?0.05). The highest level of total ammonia nitrogen (1.05 mg L^?1) and nitrite (mg L^?1) were recorded in control group, while the highest total density of heterotrophic bacteria was obtained in CN22 treatment. Growth performance parameters were at the highest level in the CN14 treatment, so the highest amount of final weight (6.88 g), biomass, and survival rate (94.79%) were observed in this treatment. The biochemical composition (protein, lipid, and ash) of the shrimp body and produced biofloc were affected by different C/N ratios, so these parameters were increased by rising of C/N ratio. In general, this study showed that the growth performance and body composition of Pacific white shrimp juveniles and water quality were suitable for rearing in biofloc system with C/N ratio of 14 than other treatments.

     

Enhancement of growth performance, digestive enzyme activities and disease resistance in black tiger shrimp (Penaeus monodon) postlarvae by potential probiotics

Outbreaks of multidrug-resistant pathogens have been constraint on aquaculture production in Thailand, thereby controlling shrimp pathogens by preventive probiotics being importance to sustain the aquaculture system. In this study, the effect of potential probiotics, Bacillus subtilis and Enterococcus sp. was related to growth, digestive enzyme activities (trypsin and chymotrypsin) and pathogenic resistance by postlarval black tiger shrimp (Penaeus monodon). The experiment was divided into two treatments, in triplicate, with and without supplementation of probiotics as a food additive. Shrimp fed with probiotics for a culture period of 84 days showed a significant increase (P < 0.05) in weight (2.03 ± 0.29 g) and survival (71.91 ± 3.15 %) in comparison with non-treated shrimp (1.53 ± 0.28 g and 65.20 ± 5.68 %, respectively). Trypsin activity of treated shrimp was significantly higher (P < 0.05) than that of control shrimp whereas chymotrypsin activities of the two treatments were not significantly different (P > 0.05). After challenging P. monodon postlarvae with a shrimp pathogen, Vibrio harveyi for 10 days, percentage of mortality of P. monodon postlarvae fed with probiotics was 46.67 ± 1.44 %, significantly lower (P < 0.05) than that of non-treated shrimp (61.67 ± 6.29 %). This study showed that potential probiotics was appropriate for application in P. monodon postlarvae cultivation under laboratory condition due to improvement of shrimp weight and survival, enhancement of trypsin activity and reduced mortality causing by pathogenic V. harveyi. This is the first publication reported the effect of probiotics on trypsin and chymotrypsin activities of P. monodon postlarvae.     

C/N ratio control and substrate addition for periphyton development jointly enhance freshwater prawn Macrobrachium rosenbergii production in ponds

The present research investigated the effect of carbon/nitrogen ratio (C/N ratio) control in ponds with or without substrate addition for periphyton development on production of giant freshwater prawn. C/N ratios of 10, 15 and 20 were investigated in 40 m^− 2 ponds stocked with 2 prawn juveniles (5.023 ± 0.02 g) m^− 2 with or without added substrates for periphyton development. The various treatment combinations of C/N ratio and periphyton substrate addition are abbreviated as ‘CN10’, ‘CN15’, ‘CN20’, ‘CN10 + P’, ‘CN15 + P’ and ‘CN20 + P’, P representing periphyton substrate. A locally formulated and prepared feed containing 30% crude protein with C/N ratio10 was applied. Tapioca starch was used as carbohydrate source for manipulating C/N ratio and applied to the water column separately from the feed. Increasing the C/N ratio from 10 to 20 reduced (P < 0.001) the total ammonia-nitrogen (TAN), nitrite–nitrogen (NO₂–N) and nitrate–nitrogen (NO₃–N) in water column and total Kjeldahl nitrogen (TKN) in sediment. The addition of substrates only influenced the NO₂–N concentration in the water column (P < 0.001). Increasing the C/N ratio raised the total heterotrophic bacterial (THB) population in the water column, sediment and periphyton (P < 0.001). It also increased the dry matter (DM), ash free dry matter (AFDM), and chlorophyll a content of periphyton (P < 0.001). The lowest specific growth rate (SGR), the highest food conversion ratio (FCR), and the lowest protein efficiency ratio (PER) were recorded in treatment CN10 (P < 0.05). The addition of substrates did not influence size at harvest (P > 0.05) but improved the survival from 62.8 to 72% (P < 0.001). Increasing the C/N ratio from 10 to 20 increased the net yield by 40% and addition of substrate increased the net yield by 23%. The combination of C/N ratio control and substrate addition increased the net yield by 75% from 309 (CN10) to 540 (CN20 + P) kg ha^− 1 (120 days)^− 1. This 75% higher production concurred with (1) a lower inorganic nitrogen content in the water column, (2) a higher THB abundance supplying additional single cell protein to augment the prawn production, and (3) an improved periphyton productivity and quality.     

Application of different doses of calcium hydroxide in the farming shrimp Litopenaeus vannamei with the biofloc technology (BFT)

The reduction in alkalinity and pH occurs due to the consumption of inorganic carbon by bacteria present in the biofloc. The objective of the study was to evaluate the effects of different doses of calcium hydroxide on the water quality and growth performance of the Litopenaeus vannamei in a biofloc system. The experiment consisted of four treatments with three repetitions for each treatment: Control treatment (TC), in which the pH maintained above 7.2 due to the application of 0.05 g L^?1 of calcium hydroxide, and treatments T10, T20 and T40, in which daily doses of calcium hydroxide equivalent to 10, 20 and 40 % of the total amount of feed offered to the shrimp, respectively, were added to the environment. Twelve experimental units (150 L) were stocked with 85 juveniles of L. vannamei (0.18 ± 0.06 g), corresponding to a density of 425 shrimp m^?3, and cultivated for 56 days. The values of alkalinity and pH for treatments TC and T10 were similar and significantly lower (p < 0.05) than those for treatments T20 and T40, which differed (p < 0.05) between each other. In terms of growth performance, TC, T10 and T20 performed significantly better (p < 0.05) than T40. There were no significant differences (p > 0.05) in the survival rates. The results obtained indicate that doses of 0.05 g L^?1 of calcium hydroxide or daily applications between 10 and 20 % of the total amount of feed offered to the shrimp can be used for the correction of alkalinity and pH.     

Effect of feeding level on growth,body composition,fatty acid profile,and nutrient accumulation in shrimp (<Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>)

A study was conducted in a recirculating aquaculture system (RAS) to assess the growth performance, body composition, fatty acid profile, and nutrient accumulation in different tissues of shrimp using two feeding levels (10 and 16 g/kg^0.8/day). The shrimp PL₂₅ was nursed in the nursing tank for 2 months and stocked in six grow out tanks (ten individuals in each 30 L tank). A commercial feed with 42.7% protein was used over a 16-h period per day. After 4 weeks, the final average weight when fed 16 g/kg^0.8/day was 9.7 g which was not significantly different from a final weight of 8.6 g achieved when feeding 10 g/kg^0.8/day. The protein and energy content increased in high feeding level whereas moisture, crude fat, and ash content decreased with feeding level (P?>?0.05). With the high feeding level, the fatty acid content increased in the eye, muscle, and whole body, with eye containing the highest concentration. The isotopic ratio of δ¹³C was enriched with feeding level in the muscle and whole shrimp whereas the ratio of δ¹³C was depleted in the viscera with feeding level. The fraction of δ¹⁵N was enriched with decreasing feeding level and highest value was found in the muscle than in the whole shrimp and viscera.     

Enriched Artemia with L-lysine and DL-methionine on growth performance,stress resistance,and fatty acid profile of Litopenaeus vannamei postlarvae

A 15-day study was conducted to evaluate the effect of enrichment of Artemia metanauplii with four different emulsions, including: (A) Easy-DHA Selco; (B) Easy-DHA Selco + L-lysine (4 g/L); (C) Easy-DHA Selco + DL-methionine (4 g/L); and (D) Easy-DHA Selco + L-lysine (2 g/L) + DL-methionine (2 g/L) on growth and stress resistance of white leg shrimp Litopenaeus vannamei larvae (from Mysis I to postlarvae12). It is noticed that shrimp PL in group B had the highest wet (36.1 ± 3.1 mg) and dry (1116.7 ± 60.0 µg) weights as well as stress resistance to fresh water (64.0 ± 5.5%).     

Water quality,phytoplankton composition and growth of Litopenaeus vannamei (Boone) in an integrated biofloc system with Gracilaria birdiae (Greville) and Gracilaria domingensis (Kützing)

A 28-day indoor trial was conducted to evaluate the water quality, phytoplankton composition and growth of Litopenaeus vannamei in an integrated biofloc system with Gracilaria birdiae and Gracilaria domingensis. The experimental design was completely randomized with three treatments: control (shrimp monoculture); SB (shrimp and G. birdiae) and SD (shrimp and G. domingensis), all with three replicates. Random sampling was done (6 % of total population per experimental unit) to confirm white spot syndrome Virus (WSSV) infection using nested-PCR analysis due to suspicion of presence of the virus in the experiment (treatment and control groups). Shrimp L. vannamei (2.63 ± 0.10 g) were stocked in experimental tanks at a density of 425 shrimp m^?3, and the Gracilaria was stocked at a biomass of 2.0 kg m^?3. Shrimp mortality began in both the experimental and control groups at 10 days of culture. The integrated biofloc system (shrimp and seaweed) increased settleable solids (by 26–52 %); final weight (by 6–21 %); weekly growth (by 17–43 %); weight gain (by 17–43 %); specific growth rate (by 16–36 %); and yield (by 5–7 %) and decreased feed conversion ratio (by 21–28 %) and Cyanobacteria density about 16 % as compared to the control (shrimp monoculture). The use of red seaweed Gracilaria in an integrated biofloc system can enhance shrimp growth and reduce Cyanobacteria density in the presence of WSSV.     

Use of shrimp protein hydrolysate in Nile tilapia (Oreochromis niloticus, L.) feeds

A 45-day feeding trial was carried out to evaluate the use of shrimp protein hydrolysate (SPH) in diets for Oreochromis niloticus, L. SPH was included in isonitrogenous diets replacing fish meal protein by 0, 5, 10, and 20% and offered to Nile tilapia juveniles (1.7 ± 0.4 g) stocked in 40-L glass aquaria. The inclusion of SPH produced no significant differences (P ≥ 0.05) in final weight, survival, weight gain, average daily gain, specific growth rate, feed conversion ratio, protein efficiency ratio, or apparent net protein utilization. The inclusion of SPH Nile tilapia diets significantly affected (P < 0.05) the final fish body composition. Protein and ash contents decreased and fat content increased slightly with the increase in SPH. This study has demonstrated that SPH is a promising protein feedstuff and could account for as much as 6% of Nile tilapia diets with no adverse effects on growth and nutrient utilization.     

Effect of temperature decrease on hemocyte apoptosis of the white shrimp Litopenaeus vannamei

This study investigated the effect of continuous temperature decrease on hemocyte apoptosis of the white shrimp Litopenaeus vannamei. In the stress group, water temperature decreased from 26 to 17 °C at a rate of 1 °C/h. Shrimp kept at 26 ± 0.5 °C were used as control group. Total hemocyte count (THC), reactive oxygen species (ROS) production, cytoplasmic free-Ca²⁺ (CF-Ca²⁺) concentration, mitochondrial membrane potential (MMP), apoptotic cell ratio, and caspase-3 activity of L. vannamei hemocytes were determined when water temperature decreased to 23, 20, and 17 °C, respectively. Increased ROS production in hemocytes was observed when water temperature decreased to 20 and 17 °C. Decreased THC and cellular MMP, increased CF-Ca²⁺ concentration, apoptotic cell ratio, and caspase-3 activity were shown when water temperature decreased to 17 °C. These results indicate that water temperature decrease can induce oxidative stress on shrimp hemocytes and then cause mitochondria and caspase-3 mediated hemocyte apoptosis and THC reduction, when water temperature decreased to an unconformable level.     

Purification and characterization of chymotrypsin from viscera of vermiculated sailfin catfish, Pterygoplichthys disjunctivus, Weber, 1991

Pterygoplichthys disjunctivus viscera chymotrypsin was purified by fractionation with ammonium sulfate (30–70 % saturation), gel filtration, affinity, and ion exchange chromatography. Chymotrypsin molecular weight was approximately 29 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), shown a single band in zymogram. Electrofocusing study suggested being an anionic enzyme (pI ≈ 3.9), exhibiting maximal activity at pH 9 and 50 °C, using Suc-Ala-Ala-Pro-Phe-p-nitroanilide (SAAPNA) as substrate. Enzyme was effectively inhibited by phenyl methyl sulfonyl fluoride (PMSF) (99 %), and N-tosyl-l-phenylalanine chloromethyl ketone (TPCK) (94 %). Enzyme activity was affected by the following ions in decreasing order: Hg²⁺, Fe²⁺, Cu²⁺, Li¹⁺, Mg²⁺, K¹⁺, Mn²⁺, while Ca²⁺ had no effect. Chymotrypsin activity decreased continuously as NaCl concentration increased (from 0 to 30 %). K _m and V _max values were 0.72 ± 1.4 mM and 1.15 ± 0.06 μmol/min/mg of protein, respectively (SAAPNA as substrate). Results suggest the enzyme has a potential application where low processing temperatures are needed, such as in fish sauce production.     

Phylogenetic analysis of bacterial communities in the shrimp and sea cucumber aquaculture environment in northern China by culturing and PCR–DGGE

In this study, we investigated the bacterial communities in the shrimp and sea cucumber culture environment, including shrimp ponds (SP), sea cucumber ponds (SCP), mixed-culture ponds (MCP) and the effluent channel (EC) in Qingdao, China. Bacteria cultivation showed that the counts of heterotrophic, nitrate-reducing and sulfate-reducing bacteria in the sediment of SP were higher than that in the sediment of SCP and MCP, varying between 8.7 × 10⁴ and 1.86 × 10⁶, 2.1 × 10⁴ and 1.1 × 10⁵, and 9.3 × 10¹ and 1.1 × 10⁴ CFU g^?1, respectively. In contrast, the counts of ammonium-oxidizing and nitrifying bacteria in the sediment of SP was lower than that in the sediment of SCP and MCP. Denaturing gradient gel electrophoresis (DGGE) of 16S rDNA gene and dendrogram analyses showed that bacterial diversity in the mixed-culture environment was higher than that in the monocultures. The similarity of bacterial community between EC and SCP or MCP was higher than that between EC and SP. These results indicated that sea cucumber culture played a significant role in influencing the environmental bacterial communities that were composed mainly of Flavobacteriaceae (64.3%), Bacteriodetes (21%) and delta proteobacteria (14.7%), including the genera of Croceimarina, Lutibacter, Psychroserpens and so on. The results explained the benefit of sea cucumber culture in shrimp ponds at the level of microbial ecology.     

Biofilm versus biofloc: Are artificial substrates for biofilm production necessary in the BFT system?

The aim of this study was to evaluate the addition of artificial substrates in biofloc technology systems that are used for the intensive production of Litopenaeus vannamei. The experiment lasted 35 days. Tanks each with a useful volume of 800 L were filled with water containing bioflocs (25 % of the total volume) and filtered seawater. Three treatments with three replicates each were compared: (1) Ctrl (control), i.e., without the addition of artificial substrate, (2) 200 and (3) 400, with added artificial substrate equivalent to 200 and 400 % of the lateral area of the tanks, respectively. The shrimp with initial weight of 0.40 ± 0.15 g were stocked at a density of 300 shrimp/m². Feed (Guabi/38 Active, Brazil) was supplied two times per day. The water quality parameters were suitable for the production of L. vannamei. The settleable solids (SS) were higher in the Ctrl treatment (82.1 ± 19 mL L^?1) and differed statistically from the 200 and 400 treatments that presented average SS values below 10 mL L^?1. The reuse of water with bioflocs accelerated nitrification in the tanks, and no significant differences were shown between tanks. The presence of biofilm did not interfere with ammonium or nitrite levels, but it did serve as a food source that optimized shrimp performance, as shown by higher final weights. More studies are needed to evaluate the circulation and mixing intensity effects of the substrates on water throughout the production cycle.     

Retention efficiency and release of nutrients in the digestive tract of larval shrimp (Penaeus japonicus Bate) using different microencapsulated diets

Retention efficiency and release of the nutrients in the digestive tract of larval shrimp (Penaeus japonicus) using the diets microencapsulated with gelatin and ethyl cellulose respectively were evaluated. The microencapsulated diets were produced using the fluidized bed coating process. 7.8 % gelatin and 4.2 % ethyl cellulose to the whole microencapsulated diet were respectively adopted as coating material. After immersion in 35 ‰ NaCl solution for 1 h, the nitrogen retention efficiency was significantly lower in the diet microencapsulated with gelatin compared with the diet microencapsulated with ethyl cellulose. Free amino acid retention efficiency of the diet microencapsulated with gelatin and ethyl cellulose was 12.9, 17.2 %, respectively. The mysis II larval shrimp (P. japonicus) 10 days after hatch were fed two different microencapsulated diets for 20 days and grew significantly more than the control larval shrimp fed with Artemia and shrimp flake. The nutrient components in intermediate intestine of larval shrimp were increased gradually in the order of the control (50 % shrimp flake + 50 % Artemia), Group I (50 % diet microencapsulated with gelatin + 25 % shrimp flake +25 % Artemia), Group II (100 % diet microencapsulated with gelatin), and Group III (100 % diet microencapsulated with ethyl cellulose). This study confirmed that the microencapsulated diets with slow and controlled release characteristic in the digestive tract of the larval shrimp (P. japonicus).     

Chemical composition and physical properties of salted shrimp paste (Kapi) produced in Thailand

Chemical composition and physical properties of 11 salted shrimp pastes (Kapi) obtained from various places of Thailand were determined. Based on proximate composition, protein constituted the major component (29.44–53.27 %, dry wt. basis). All samples contained 22.77–35.47 % NaCl with A _w of 0.695–0.774. Various formal nitrogen contents (11.96–22.87 mg N/g sample) were in agreement with different degrees of hydrolysis (12.68–20.76 %), suggesting the varying cleavage of peptides among the samples. From electrophoretic study, salted shrimp paste contained a large amount of small molecular weight proteins and peptides. Different samples had different colors with \( \Delta E^{*} \) of 47.10–60.43 and \( \Delta C^{*} \) of 9.46–20.76. The samples had total carotenoid content of 0.54–1.97 mg/g sample. Free astaxanthin, astaxanthin diester and canthaxanthin were the major carotenoids in salted shrimp paste. Thus, salted shrimp paste is a good source of protein and serves as the nutritious condiment.     

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.     

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.     

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.