Effects of formaldehyde on nitrification in biofilters of small‐scale recirculating systems

Florfenicol (Aquaflor®) is the only U.S. Food and Drug Administration (FDA) approved drug for treating diseased fish reared in recirculating aquaculture systems (RAS). Treating diseased fish in RAS is challenging because of the potential to damage nitrifying bacteria in the biofilters. Impaired nitrification can lead to concentrations of ammonia and nitrite that compromise fish welfare. The objective of this study was to determine the effects of a FDA‐approved parasiticide and fungicide, Parasite‐S^® (formalin), on biofilter nitrification. Stable biofilters were exposed once to 0, 9.25, 18.5, 37, or 55.5 mg/L formaldehyde. Total ammonia nitrogen (TAN) and nitrite nitrogen were monitored daily before and throughout the study to quantify biofilter function. Formaldehyde concentrations ≥37 mg/L increased TAN and nitrite nitrogen concentrations, and nitrification did not recover to pre‐exposure concentrations up to 8 day postexposure. On the basis of those results, a second trial was conducted. Stable biofilters were exposed once or on four consecutive days to 9.25 or 18.5 mg/L formaldehyde. Biofilters repeatedly exposed to formaldehyde showed signs of impairment and had variable recovery relative to single exposures. Results of this study may help identify formaldehyde concentrations that can be safely applied to RAS when treating diseased fish.     

Effect of water velocity on ammonium and nitrite removal in pilot scale fixed bed biofilters

The effect of water velocity on nitrification rates in fixed bed biofilters was investigated in three freshwater pilot scale RAS with rainbow trout. Removal of total ammonia nitrogen (TAN) and nitrite-nitrogen were assessed by NH₄Cl spikes and tested at four different water velocities in the biofilters (1.4, 5.4, 10.8 and 16.2 m h⁻¹) under identical conditions. Water velocities below 10.8 m h⁻¹ significantly reduced TAN- and nitrite removal rates. The surface specific TAN removal rates correlated with the TAN concentrations at the water velocities 10.8 and 16.2 m h⁻¹, and the first order surface removal rate constant was estimated at 0.45 m h⁻¹. However, no correlations between TAN removal and TAN concentrations were found at the lowest velocities. Up to five-fold elevated nitrite levels were found in the RAS when biofilters were operated at 1.4 m h⁻¹ compared to the trials at other water velocities, substantiating the significant effect of water velocity on both nitrification processes. The importance of biofilter hydraulics documented in this pilot scale RAS probably have implications for design and operation in larger scale RAS.     

Limited water exchange production systems for freshwater ornamental fish

Two biofilter designs and a control were tested in triplicate to determine if inexpensive bioremediation devices could increase production and decrease water use on ornamental fish farms in Hawaii. Koi (Cyprinus carpio L.) were used as the model species and the experiment was conducted outdoors in greenwater. When fish density was 9.7 kg per 2.08 m³ and they were eating 125 g day^?1, the 20 L trickle filters were able to maintain acceptable water quality. Tanks with the same size submerged filters suffered significantly lower dissolved oxygen levels compared with tanks with trickle filters and control tanks with no biofilters exhibited significantly higher nitrite‐nitrogen (about 20 mg L^–1) and nitrate levels (about 400 mg L^–1). As typical ornamental fish weigh 3 g, the trickle biofilter system described here can produce 1.55 fish L^?1 (compared with the industry standard 0.25 fish L^?1) and use very little water other than the water originally in the tanks.     

A case study of biofilter activation and microbial nitrification in a marine recirculation aquaculture system for rearing Atlantic salmon (Salmo salar L.)

Marine recirculation aquaculture system (RAS) is a prominent technology within fish farming. However, the nitrifying bacteria in the biofilter have low growth rates, which can make the biofilter activation a long and delicate process with periods of low nitrification rates and variations in water quality. More knowledge on the microbial development in biofilters is therefore needed in order to understand the rearing conditions that favour optimal activation of the biofilters. In this case study, we investigated the activation of two biofilters in a marine RAS for Atlantic salmon post‐smolt associated with either high or low stocking densities of fish by monitoring the microbial communities and chemical composition. The results showed that the microbial communities in both biofilters were similar during the first rearing cycle, despite variations in the water quality. Nitrifying bacteria were established in both biofilters; however, the biofilter associated with low stocking density had the highest relative abundance of ammonia‐oxidizing Nitrosococcus (1.0%) and nitrite‐oxidizing Nitrospira (2.1%) at the end of the first rearing cycle, while the relative abundance of ammonia‐oxidizing Nitrosomonas (2.3%–2.9%) was similar in both biofilters. Our study showed that low fish stocking density during the first rearing cycle provided low and steady concentrations of ammonium, nitrite and organic load, which can stimulate rapid development of a nitrifying population in new marine RAS biofilters.     

Peracetic acid degradation and effects on nitrification in recirculating aquaculture systems

Peracetic acid (PAA) is a powerful disinfectant with a wide spectrum of antimicrobial activity. PAA and hydrogen peroxide (HP) degrade easily to oxygen and water and have potential to replace formalin in aquaculture applications to control fish pathogens, for example the ectoparasite, Ichthyophthirius multifiliis.We studied water phase PAA and HP decay in three aquaculture situations, i) batch experiments with two types of system waters, ii) PAA decay at different fish densities, and iii) degradation of PAA in submerged biofilters of recirculating aquaculture systems (RAS). Furthermore, effect of PAA on the nitrification activity and the composition of the nitrifying population were investigated.PAA and HP decay showed first order kinetics. High dosage PAA/HP in water with low COD inhibited HP removal, which was not observed in water having a higher COD content. PAA decay was significantly related to fish stocking density, with half life constants for PAA of 4.6 and 1.7 h at 12 and 63 kg m^− 3, respectively.PAA application to RAS biofilter showed rapid exponentially decay with half life constants of less than 1 h, three to five times faster than the water phase decay rates.Biofilter surface specific PAA removal rates ranged from 4.6 to 13.9 mg PAA m^− 2 h^− 1 and was positively correlated to the nominal dosage. Low PAA additions (1.0 mg L^− 1) caused only minor impaired nitrification, in contrast to PAA application of 2.0 and 3.0 mg L^− 1, where nitrite levels were significantly increased over a prolonged period, albeit without fish mortality. The dominant ammonium oxidizer was Nitrosomonas oligotropha and the dominant nitrite oxidizer was Nitrospira. Based on the present findings and other recent results from field and in vitro studies, application perspectives of PAA are discussed.     

Microbial dynamics in RAS water: Effects of adding acetate as a biodegradable carbon-source

This study evaluated the effect of an abrupt increase in easily biodegradable carbon (acetate) on bacterial activity and abundance in the water of recirculating aquaculture systems (RAS). The study included a batch experiment with RAS water only, and an experiment at system level where twelve pilot scale RAS were used. The batch experiment was made to test how acetate concentration would influence the microbial state in RAS water. Further, we wanted to observe if the selected microbial analysis tools would be able to detect these changes. The second experiment was carried out in twelve identical and independent RAS that had been operated under constant loading conditions (1.6 kg/m³ make-up water) for five months prior to the trial. The twelve RAS were divided into four treatment groups in triplicates: i) control with submerged biofilter (Ctrl + bf); ii) control without submerged biofilter (Ctrl-bf); iii) acetate addition in RAS with submerged biofilter (Ac + bf); and iv) acetate addition in RAS without submerged biofilter (Ac-bf). The biofilter media from the groups without submerged biofilter (Ac-bf and Ctrl-bf) was removed just 5 h prior to the start of the trial. The two acetate treatment groups (Ac + bf and Ac-bf) were spiked with 40 mg/L of acetate three consecutive times (0, 24 and 48 h). Consumption of acetate, bacterial abundance and bacterial activity were followed for 72 h after the first acetate spike for both experiments. Bacterial activity was quantified by BactiQuant^® and hydrogen peroxide (HP) degradation assay. Bacterial abundance was assessed by quantifying micro-particles and free-living bacteria. In the batch experiment we observed a significant increase in bacterial activity proportional to the amount of acetate added, and a corresponding significant increase in microparticles (1–3 μm). In the pilot scale RAS experiment, the acetate addition in RAS with submerged biofilter did not cause an increase in bacterial activity, or in the number of microparticles in the water phase but a significant increase in bacterial activity and number of microparticles were observed in the RAS without submerged biofilter (Ac-bf). These changes were particularly pronounced shortly after each acetate spike.In RAS with submerged biofilters, the acetate was presumably consumed primarily by the bacterial community within the biofilm, and consequently, only minor changes were observed in densities of free-living bacteria in the water phase. The results of the study suggest that heterotrophic bacteria in the submerged biofilter have a high capacity to handle fluctuation of organic matter loading in RAS, thereby stabilizing the abundance and activity of bacteria in the water column.     

Effects of abrupt salinity increase on nitrification processes in a freshwater moving bed biofilter

The nitrification process is a widely used biological approach responsible for ammonia and nitrite removal in recirculating aquaculture system (RAS) biofilters. Given this pivotal role, the influence of different water quality parameter on nitrification efficiency is important information for RAS operations. One influencing parameter is salinity, and salinity fluctuations in freshwater RAS biofilters are reported to affect the nitrifying bacteria. This study investigated the effects of abrupt increase in salinity in freshwater RAS on substrate-dependent (1’-order) as well as substrate independent (0’-order) nitrification rates. A 100% inhibition was found for surface specific removal (STR) of total ammonia nitrogen (TAN) and surface specific nitrite removal (SNR) when salinity was abruptly increased to 25‰ and above. A fast turnover (i.e. steep decline in [NH₄-N⁺] and [NO₂-N⁻]) were observed at lower salinities (≤10‰), while limited/no degradation of either ammonia or nitrite was seen at salinities above 25‰. At low substrate loading (1’-order process), removal rate constants (k_1a) of 0.22 and 0.23 m d^-1 were observed for ammonia and nitrite degradation, respectively, declining to 0.01 m d^-1when adding marine RAS water increasing the salinity to 15‰. Similar observations followed at high nutrient loadings (0’-order process) with STR and SNR of 0.10 and 0.12 g N m^-2 d^-1, respectively, declining to 0.01 g N m^-2 d^-1 at 15‰. When salinities of 25‰ and 35‰ were applied, neither TAN nor nitrite degradation was seen. The results thus demonstrate a pronounced effect of salinity changes when freshwater RAS biofilters are subjected to fast/abrupt changes in salinity. RAS facility operators should be aware of such potential effects and take relevant precautions.     

Depletion of florfenicol amine in tilapia (Oreochromis sp.) maintained in a recirculating aquaculture system following Aquaflor®‐medicated feed therapy

Aquaflor^® [50% w w^?1 florfenicol (FFC)], is approved for use in freshwater‐reared warmwater finfish which include tilapia Oreochromis spp. in the United States to control mortality from Streptococcus iniae. The depletion of florfenicol amine (FFA), the marker residue of FFC, was evaluated after feeding FFC‐medicated feed to deliver a nominal 20 mg FFC kg^?1 BW d^?1 dose (1.33× the label use of 15 mg FFC kg^?1 BW d^?1) to Nile tilapia O. niloticus and hybrid tilapia O. niloticus × O. aureus held in a recirculating aquaculture system (RAS) at production‐scale holding densities. Florfenicol amine concentrations were determined in fillets taken from 10 fish before dosing and from 20 fish at nine time points after dosing (from 1 to 240 h post‐dosing). Water samples were assayed for FFC before, during and after the dosing period. Parameters monitored included daily feed consumption and biofilter function (levels of ammonia, nitrite and nitrate). Mean fillet FFA concentration decreased from 13.77 μg g^?1 at 1‐h post dosing to 0.39 μg g^?1 at 240‐h post dosing. Water FFC concentration decreased from a maximum of 1400 ng mL^?1 at 1 day post‐dosing to 847 ng mL^?1 at 240 h post‐dosing. There were no adverse effects noted on fish, feed consumption or biofilter function associated with FFC‐medicated feed administration to tilapia.     

Degradation of urea,ammonia and nitrite in moving bed biofilters operated at different feed loadings

This study investigated how removal rates of urea, ammonia, and nitrite in laboratory scale moving bed biofilters were affected by long-term feed loading. To generate different loadings, five identical freshwater flow-through systems (100 l/h) with rainbow trout (Oncorhynchus mykiss) were fed increasing fixed rations of a commercial diet. The filtered effluent from each system was lead through a moving bed biofilter installed end-of-pipe. After an acclimatization period of four months, the moving bed biofilters were spiked separately with urea, ammonia and nitrite in batch mode in three successive trials to investigate degradation kinetics. Results showed that urea, in addition to ammonia and nitrite, was degraded although the substrate limited/dependent removal rate of urea (first order kinetic) was lower than that of ammonia and nitrite. Degradation of urea could be described as first order kinetics below 2.5 mg N/l. Degradation of total ammonia nitrogen (TAN) and nitrite was substrate independent (zero order kinetic) above 2 mg N/l and subsequently substrate dependent as substrate concentrations in the bulk water declined. The transition zone from zero to first order degradation was elevated with increase in long-term biofilter loading. For ammonia and nitrite, a significant increase in the zero order removal rate constants related to long-term loading were observed up to a long-term feed loading of 207 g/d, corresponding to 69 g feed/m² filter media/d and an TAN + urea-N concentration of 2.70 mg N/l. Long-term feed loading had no obvious effect on first order removal rate constants of any of the three nitrogenous compounds. Degradation of urea resulted in generation of ammonia demonstrating that urea degradation contributes to the ongoing nitrification activity in aquaculture biofilters. For all three types of spiking (urea, ammonia and nitrite) accumulation of nitrate was observed in the moving bed biofilters, sustaining that nitrification had occurred.     

Use of fluidized bed biofilter and immobilized Rhodopseudomonas palustris for ammonia removal and fish health maintenance in a recirculation aquaculture system

Intensive recirculating aquaculture relies on biofilters to sustain satisfactory water quality in the system. Fluidized bed and immobilized cell technologies were used to remove ammonia from the water and maintain fish health. A high‐rate nitrifying fluidized bed biofilter combined with valveless filter was designed for use in a recirculation aquaculture system (RAS). The suspended solids produced during fish culture could automatically be removed using a valveless filter. Natural porosity with fitting proportion, steady fluidization and expanding rate was chosen as the fluidized carrier. The technology of bacterial separation and cultivation was used. The immobilized Rhodopseudomonas palustris (R. palustris) produced through a biotechnologically embedding medium is suitable for fish and could help prevent diseases. Nitrification was promoted through the selective rearing of nitrobacteria in a fluidized bed biofilter. Water quality was improved using fluidized bed biofilter and immobilized R. palustris in the RAS. In addition, the proposed system was able to reduce costs. Maximum fish load was 45 ± 3 kg m^?3 in the closed recirculating water fish culture system, and water use was reduced by 80–90%. The total ammonia nitrogen removal rate of the technology was 80–95%, and nitrite N removal rate was above 80%.     

Optimization of blue mussel (Mytilus edulis) seed culture using recirculation aquaculture systems

By introducing recirculation aquaculture systems (RAS) in the nursery phase of the blue mussel (Mytilus edulis) (17–18 mm), we aimed at a similar growth and survival and a similar water quality compared to the commonly used flow‐through systems (FTS). To calculate water flow and size of the biofilter, a series of experiments were done to determine clearance rate (9.26 mL min^?1), pseudo faeces threshold (60 000 cells Pavlova lutheri mL^?1), nitrogen production (0.00065 mg TAN h^?1 ind^?1 and 1.6 × 10^?5 mg NO₂–N h^?1 ind^?1) and oxygen consumption (0.03 ± 0.01 mg O₂ h^?1 ind^?1). RAS showed no significant differences in water quality (0.06 mg TAN L^?1; 7.7 mg O₂ L^?1) and growth performance of mussel seed specific growth rate (SGR = 5% day^?1) after the experimental period of 4 weeks compared with FTS. The low water refreshment, 10% per day, as well as the constant chlorophyll concentrations (9.76 ± 1.06 μg L^?1), suggests the potential of RAS as culture system for mussel seed.     

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.     

Taxonomic and functional profiling of nitrifying biofilms in freshwater,brackish and marine RAS biofilters

In recirculating aquaculture systems (RAS), the crucial step of eliminating toxic N compounds like ammonia and nitrite is mediated via nitrifying microorganisms and takes place in biofilters. In this study, analyses of microorganisms colonizing biocarriers of nine moving-bed biofilters of three different RAS operated with freshwater, brackish or marine process water uncovered site specific communities. Illumina-based amplicon sequencing of the V4-region of the 16S rRNA gene revealed a high microbial diversity with 1000–2500 species-level operational taxonomic units (OTUs) in all biofilters with the highest diversity in the brackish RAS. Proteobacteria, Bacteriodetes, Plantomycetes, Chloroflexi and Nitrospirae represented the most abundant phyla. 76 out of 674 known genera occurred in all nine biofilters and were defined as core-taxa, including nitrifying bacteria (Nitrosomonas and Nitrospira) as well as members of the (heterotrophic) genera Planctomyces, Blastopirellula, Nannocystis and Lewinella. Nitrifying communities composed of different, closely related and so far uncultured members of Nitrosomonas and Nitrospira were identified, strongly indicating that several potentially novel ammonia and nitrite oxidizing species are present in RAS biofilters. Relatives of known comammox Nitrospira were detected in the brackish biofilters, revealing 94–99 % identity of the 16S rRNA gene sequence to Ns. inopinata. Salinity tolerance tests with biocarriers derived from biofilters of the three distinct RAS showed an unexpected broad physiological flexibility with regard to salinity. Nitrification performance of freshwater nitrifiers was drastically reduced with increasing salinity and nearly completely inhibited at 15 PSU, while the brackish and marine nitrifiers showed a high resistance and maintained nitrification activity in a broad range of salt concentrations. This data can help to improve the nitrification process in RAS with changing salinity of the process water.     

Startup and effects of relative water renewal rate on water quality and growth of rainbow trout (Oncorhynchus mykiss) in a unique RAS research platform

The aquaculture industry is growing fast but facing two major challenges: a shortage of suitable locations for growth and the need to reduce environmental impacts. One solution for both these challenges is inland production through recirculating aquaculture systems (RAS). The RAS technique is rather new, and several practical issues need to be solved. In this study, an experimental platform, consisting of ten individual RAS units, was built for small-scale testing of different RAS designs and operation methods, and two preliminary experiments were conducted. In the first experiment, the capability of different chemical additions (sodium nitrite, ammonium chloride and/or cane sugar) to fasten the startup of the nitrification bioreactor was tested. In addition, the suitability and reliability of an online water measurement system in monitoring nitrification process with was evaluated. We demonstrated that when using a combination of sodium nitrite and ammonium chloride in a concentration of 5 mg l⁻¹, nitrification started one week before than when using only ammonium chloride or a clean start with rainbow trout (Oncorhynchus mykiss). In the second experiment, the effect of different relative water renewal rates (RWR) on water quality, rainbow trout growth and feed conversion ratio (FCR) were examined at 16 °C. Based on the results, FCR increased when RWR went below 478 l kg⁻¹, and the specific growth rate decreased when RWR went below 514 l kg⁻¹. Furthermore, when RWR decreased, nitrate, nitrite and organic material accumulated in the circulating water. In conclusion, we showed using experimental RAS platform that online water quality monitoring is a useful tool in following the effect of different management practices. Furthermore, we demonstrated that chemical substrate additions provide the fastest biofilter startup, and that water management is still in the key role in defining the fish production in RAS.     

Pyocyanin as a safe aquaculture drug for the control of vibriosis in shrimp recirculating aquaculture system (RAS)

Diseases are one of the most critical limiting factors in aquaculture. Recirculating aquaculture systems (RAS) are one of the most functionally viable sustainable aquaculture production systems adopted world over. In the event of any eventuality caused by bacterial pathogens, antibiotics or other antibacterial agents cannot be applied due to the vulnerability of biological filters which form an integral part of the RAS. Because of this, newer drugs are required for the effective control of diseases in RAS which would not interfere with the activity of nitrifying bacteria used in the biological filters. The antagonistic activity of pyocyanin, a bioactive compound produced by Pseudomonas aeruginosa, against vibrios is well established. The purpose of this study was to prove the effectiveness of pyocyanin as an aquaculture drug for application in RAS by employing a pilot-scale shrimp culture under RAS. It was noted that at the concentration of 5 mg L^?1, pyocyanin could significantly bring down the population of Vibrio spp. in RAS without affecting noticeably the other natural heterotrophic bacteria. Also, pyocyanin at this concentration did not inhibit the activity of nitrifying bacterial consortia used in the SBSBR (stringed bed suspended bioreactor) of RAS. The reared shrimp (Penaeus monodon) showed 100% survival after the application of pyocyanin, besides exhibiting normal health signs. Pyocyanin was non-toxic to the shrimp hybrid cell line (PmLyO-Sf9) at the concentration required for its application in disease management (IC₅₀?=?419.26 mg L^?1). The present study has demonstrated that pyocyanin is effective as an environment-friendly and safe aquaculture drug for the application in RAS-based shrimp culture to control Vibrio spp. without impeding nitrification established through the deployment of nitrifying bioreactors.

     

Effects of ammonia and nitrite on survival, growth and moulting in juvenile tiger crab, Orithyia sinica (Linnaeus)

The effects of ammonia and nitrite on survival, growth and moulting were investigated in juvenile tiger crab, Orithyia sinica (carapace length 3.91±0.15 mm, carapace width 3.84±0.23 mm, n=440), after 30 days exposure to ammonia‐N (0, 20, 50, 100 and 150 mg L^?1) and nitrite‐N (0, 50, 100, 150, 200 and 250 mg L^?1) using a continuous flow system. Survival rates of tiger crab exposed to ammonia and nitrite decreased linearly with the exposure time and concentration. The growth rate of tiger crab exposed to 50, 100 and 150 mg L^?1 ammonia was significantly lower than that of control crabs. The growth rate of tiger crab exposed to nitrite decreased at 150, 200 and 250 mg L^?1 nitrite. During the ammonia and nitrite exposure, the intermoult period of the juveniles of tiger crab O. sinica was shortened between the first and second moult, and the number of moulting of crabs exposed to a higher concentration were significantly higher than that of control crabs.     

Nutrient removal efficiency of Hydropuntia cornea in an integrated closed recirculation system with pink shrimp Farfantepenaeus brasiliensis

In this study, we have tested the effect of seaweed stocking density in an experimental seaweed biofilter using the economically important red seaweed Hydropuntia cornea integrated with the cultivation of the pink shrimp Farfantepenaeus brasiliensis. Nutrient removal efficiency was evaluated in relation to seaweed stocking density (2.5, 4, 6 and 8 g fw L^?1). Total ammonia nitrogen (TAN) was the main nitrogen source excreted by F. brasiliensis, with concentrations ranging from 41.6 to 65 μM of NH₄⁺‐N. H. cornea specific growth rates ranged from 0.8 ± 0.2 to 1.4 ± 0.5% day^?1 with lowest growth rates at higher seaweed stocking density (8 g fw L^?1). Nutrient removal was positively correlated with the cultivation densities in the system. TAN removal efficiency increased from 61 to 88.5% with increasing seaweed stocking density. Changes in the chemical composition of the seaweed were analysed and correlated with nutrient enrichment from shrimp effluent. The red seaweed H. cornea can be cultured and used to remove nutrients from shrimp effluents in an integrated multi‐trophic aquaculture system applied to a closed recirculation system. Recirculation through seaweed biofilters in land‐based intensive aquaculture farms can also be a tool to increase recirculation practices and establish full recirculation aquaculture systems (RAS) with all their known associated benefits.     

Hydrogen peroxide oxygenation and disinfection capacity in recirculating aquaculture systems

Hydrogen peroxide (H₂O₂) treatment is an alternative for disinfection in aquaculture, which may be advantageous as it dissociates and disinfects while increasing water oxygen concentration. Yet, accurate dosing remains undeveloped in Recirculating Aquaculture Systems (RAS). Dosage requirements can depend on organic burden, stocking density, feeding frequency, salinity, temperature and biofilter performance. The present case study investigated the dual effect of H₂O₂ application for oxygen enrichment and disinfection when continuously applied to a RAS rearing European seabass. H₂O₂ addition equivalent to 2.4 and 15.8 H₂O₂ mg L⁻¹ were applied for 4 h per day in three 5-days experiments. H₂O₂ was injected at the inlet of protein skimmer and/or the rearing tanks in or without combination with traditional disinfection methods. Water microbial load and oxygen saturation were determined, along with stress markers glucose and cortisol in blood plasma of fish. Doses of 15.8 mg L⁻¹ H₂O₂ steadily increased oxygen levels in holding tank water from ∼50 % to over 100 % saturation while reducing microbial load (from 604.4 CFU ml⁻¹ in the rearing tanks before dosing to 159.8 CFU ml⁻¹ after application), achieving suitable conditions for commercial fish densities in RAS. The doses used had negligible impact on biofilter performance and did not affect the fish in terms of stress. Overall results indicate H₂O₂ is effective for disinfection and oxygenation of RAS systems when applied at appropriate dosage and we recommend the protein skimmer as the safest position in order to protect the bacterial community of the biofilters and the reared fish.     

Haematological and biochemical profiles of carp blood following nitrite exposure at different concentrations of chloride

Haematological parameters of 2‐year‐old carp (Cyprinus carpio L.) were assessed to study the protective effect of chloride on the health of fish exposed to elevated nitrite concentrations. Four groups of carp were exposed to different concentrations of nitrite and chloride for 96 h (group E1: 67 mg L^?1 NO₂^?, 11 mg L^?1 Cl^?; group E2: 67 mg L^?1 NO₂^?, 100 mg L^?1 Cl^?; group E3: 0 mg L^?1 O₂^?, 100 mg L^?1 Cl^? and group C: 0 mg L^?1 NO₂^?, 11 mg L^?1 Cl^?). The main haematological response of carp to an acute exposure to nitrite (group E1) was a significant decrease (P<0.05) in haemoglobin concentrations (53.40±6.61 g L^?1), haematocrit (0.21±0.02 LL^?1), erythrocyte count (1.13±0.12 TL^?1), leucocyte count (7.1±4.19 GL^?1) and lymphocyte count (5.28±2.51 GL^?1), and a significant increase in methaemoglobin concentration (90.50±4.38%, P<0.01) and mean corpuscular haemoglobin concentration (0.27±0.2 LL^?1, P<0.05). At higher chloride concentrations (group E2), a lower nitrite toxicity was observed. In group E2 carp, methaemoglobin made up 38.32±13.30%. Erythrocytes in carp exposed to nitrite showed qualitative changes. Compared with the control group C, group E1 carp showed a significantly higher number (P<0.05) of elongated erythrocytes, with the nucleus located at one cell pole (0.519±0.388 TL^?1). All erythrocytes of group E1 carp had remarkably clear cytoplasms compared with the cytoplasm in the control group C. The biochemical values found were comparable with those found in controls. The main histological lesions were found in the gills of carp exposed to nitrite and consisted of hyperplasia and an elevated number of chloride cells.     

A novel approach for ammonia removal from fresh-water recirculated aquaculture systems,comprising ion exchange and electrochemical regeneration

A new physico-chemical process for ammonia removal from fresh-water recirculated aquaculture systems (RASs) is introduced. The method is based on separating NH₄⁺ from RAS water through an ion-exchange resin, which is subsequently regenerated by simultaneous chemical desorption and indirect electrochemical ammonia oxidation. Approach advantages include (1) only slight temperature dependence and no dependence on bacterial predators and chemical toxins; (2) no startup period is required and the system can be switched on and off at will; and (3) the fish are grown in much lower bacterial concentration, making the potential for both disease and off-flavor, lower. A small pilot scale RAS was operated for 51 d for proving the concept. The system was stocked by 105 tilapia fish (initial weight 35.8 g). The fish, which were maintained at high TAN (total ammonia nitrogen) concentrations (10–23 mgN L⁻¹) and fish density of up to 20 kg m⁻³, grew at a rate identical to their established growth potential. NH_3(aq) concentrations in the fish tank were maintained lower than the assumed toxicity threshold (0.1 mgN L⁻¹) by operating the pond water at low pH (6.5–6.7). The low pH resulted in efficient CO₂ air stripping, and low resultant CO_2(aq) concentrations (<7 mg L⁻¹). Due to efficient solids removal, no nitrification was observed in the fish tank and measured nitrite and nitrate concentrations were very low. The system was operated successfully, first at 10% and then at 5% daily makeup water exchange rate. The normalized operational costs, calculated based on data derived from the pilot operation, amounted to 28.7 $ cent per kg fish feed. The volume of the proposed process was calculated to be ∼13 times smaller than that of a typical RAS biofilter. The results show the process to be highly feasible from both the operational and economical standpoints.