Recirculating Aquaculture Systems Water Research Articles

Nutrient removal and biomass production of marine microalgae cultured in recirculating aquaculture systems (RAS) water with low phosphate concentration

This study was conducted to investigate the feasibility of microalgal biomass production and nutrient removal from recirculating aquaculture systems (RAS) water (RASW) with low phosphate concentration. For this purpose, Nannochloropsis oculata, Pavlova gyrans, Tetraselmis suecica, Phaeodactylum tricornutum, and their consortium were cultivated in RASW and RASW supplemented with vitamins (+V). Among them, N. oculata showed the maximum biomass production of 0.4 g/L in RASW. Vitamins supplementation significantly increased the growth of T. suecica from 0.16 g/L in RASW to 0.33 g/L in RASW + V. Additionally, T. suecica showed the highest nitrate (NO3–N) removal efficiency of 80.88 ± 2.08 % in RASW and 83.82 ± 2.08 % in RASW + V. Accordingly, T. suecica was selected for scaling up study of microalgal cultivation in RASW and RASW supplemented with nitrate (RASW + N) in 4-L airlift photobioreactors. Nitrate supplementation enhanced the growth of T. suecica up to 2.2-fold (day 15). The fatty acid nutritional indices in T. suecica cultivated in RASW and RASW + N showed optimal polyunsaturated fatty acids (PUFAs)/saturated fatty acid (SFAs), omega-6 fatty acid (n-6)/omega-3 fatty acid (n-3), indices of atherogenicity (IA), and thrombogenicity (IT)). Overall, the findings of this study revealed that despite low phosphate concentration, marine microalgae can grow in RASW and relatively reduce the concentration of nitrate. Furthermore, the microalgal biomass cultivated in RASW consisting of pigments and optimal fatty acid nutritional profile can be used as fish feed, thus contributing to a circular bioeconomy.

Plant nutritional value of aquaculture water produced by feeding Nile tilapia (Oreochromis niloticus) alternative protein diets: A lettuce and basil case study

Societal Impact StatementRefining circular multitrophic food production methods, which integrate plant, fish, and insect outputs, is imperative for environmental sustainability. Our findings suggest that the right protein choices in fish feed, like black soldier fly meal and poultry meal, can notably enhance the nutrient profile of fish waste water. This, in turn, is conducive for hydroponic cultivation, enhancing the nutritional attributes of plants like basil and lettuce. As we reduce environmental impact and optimize resource use, it is evident that our food ecosystems are deeply intertwined. Harnessing these synergies could redefine our approach to food production, paving the way for a more sustainable global future.Summary Optimization of nutrient use efficiencies in circular multitrophic food production systems (i.e., plant, fish, and insect production) is crucial for sustainability. This study tested how protein ingredient choice in fish feed influences the plant nutritional value of the fish waste water when used for hydroponic crop production. Waste water samples were obtained from recirculating aquaculture systems (RAS) in which Nile tilapia (Oreochromis niloticus) were fed different single protein source diets—black soldier fly meal (BSFM), poultry by‐product meal (PM), poultry blood meal (PBM), and fish meal. Water was analyzed for plant nutrients and used for lettuce and basil cultivation to evaluate their suitability for hydroponic crop production—viz. yield, mineral nutrient, and selected secondary metabolite levels. BSFM RAS water had the highest concentrations of K, Mg, and micronutrients (Cu, Mn, Mo, Zn) of the RAS waters, whereas PM RAS water contained the highest P concentration and had a mean pH of 6.5 closer to the optimum pH for hydroponic plants. These RAS waters consequently lead to the highest yields in basil and lettuce indicating the importance of the aforementioned factors. From a plant production perspective, BSFM appears promising as a protein source in fish feeds for aquaponics. Usage of RAS waters for plant production helps reduce (i) environmental impact of RAS water and (ii) resource input in plant production. Longer term RAS trials should be conducted to determine the maximum nutrient concentrations achievable during fish production with diets including BSFM as the main protein source.

Effect of light wavelengths on algal-bacterial symbiotic particles (ABSP): Nitrogen removal, physicochemical properties, community structure

The accumulation of nitrogenous waste, such as ammonia, and its biological nitrification products, namely nitrite and nitrate, poses one of the primary challenges in the water treatment process for Recirculating Aquaculture Systems (RAS). This study proposed the incorporation of light and algal-bacterial symbiotic particles (ABSP)into RAS as a viable solution to address these challenges. The findings indicated that ABSP achieved the RAS water quality standards (ammonia nitrogen <0.5 mg/L, nitrite nitrogen <0.1 mg/L) by the 14th day of operation, while red light achieved and maintained these standards from as early as the 9th day. Further analysis of chemical stoichiometry and functional genes revealed that microalgae (32.7%, 15.6%, 0%) and bacteria (47.7%, 55.8%, 48.4%) were responsible for the assimilation and removal of most organic and nutrient substances under red light, white light, and dark conditions, respectively. Additionally, it was observed that the presence of algae and bacteria in ABSP was significantly influenced by the wavelength of light. Under red light, an optimal distribution of functional species for both microalgae (Characium, Microcoleus, and Uronema) and bacteria (Nakamurella and Micropruina) was achieved, which is crucial for the self-sustaining synergistic reaction and stability of ABSP. This study may offer a promising engineering alternative with enormous potential to achieve energy-efficient and environmentally sustainable RAS water treatment.

Peracetic acid mode-of-action on aquaculture microbes evaluated by dual-staining flow cytometry

Flow cytometry with dual staining allows an exact quantification of live and dead cells. It is a potential tool for gaining new information on the mode-of-action of disinfection chemicals and supporting efficient microbial control in recirculating aquaculture systems (RAS). Here, we used a dual-staining flow cytometry to examine the disinfection efficiency of peracetic acid (PAA) against i) the fish pathogenic bacterium Aeromonas salmonicida and ii) on microbes present in RAS. The method allowed us to accurately quantify the A. salmonicida cell response to different PAA concentrations (1, 2, and 10 mg/L) in time and to calculate the PAA concentration needed to achieve 95% cell mortality (LC95 estimates). With the high PAA dose of 10 mg/L, LC95 was reached in <15 min, while with the intermediate dose of 2 mg/L, LC95 was reached only in the end of the 60 min trial and with the low 1 mg/L dose, the disinfection efficiency remained low. The addition of soluble organic matter increased the PAA decay and decreased the disinfection efficiency. In RAS water, disinfection efficiency increased with increasing PAA concentration (from 1 to 10 mg/L) but was significantly reduced with increasing organic matter concentration (TCOD from 7.6 to 30.3 mg O2/L). The increase in the proportion of dead cells was found to be a reliable proxy for disinfection efficiency in RAS water. In the pure cell culture, both the increase in the proportion of dead cells and the fraction of reduced live cells reflected the disinfection efficiency of PAA. Altogether, the study demonstrated the PAA mode-of-action on aquaculture microbes and showed that flow cytometry with dual staining allows fast and precise examination of microbial viability under different conditions.

Influence of dietary phosphorus on orthophosphate accumulation in recirculating aquaculture systems with rainbow trout (Oncorhynchus mykiss)

Dietary phosphorus (P) levels affect the excretion of orthophosphate (PO4-P) from fish. Since most recirculating aquaculture systems (RAS) do not specifically remove PO4-P within the recirculating loop, dietary P levels may influence PO4-P concentrations in RAS water. A feeding trial was performed in freshwater RAS, aiming to determine the relationship between dietary P levels and PO4-P accumulation in RAS water, and to examine potential effects of dietary P levels on microbial activity and abundance in RAS water given that P is an essential mineral for all microorganisms. Three diets with 1.49% (HiP), 1.20% (MeP), and 0.89% (LoP) total P, respectively, were each fed to triplicate pilot-scale RAS with juvenile rainbow trout (Oncorhynchus mykiss) for five weeks, applying a daily feed loading of 1.67 kg feed/m3 make-up water. Digestible dietary P levels correlated significantly with the PO4-P concentrations in RAS water at the end of the trial, while no similar correlations with microbial abundance or activity were observed. Nitrate-N concentrations measured by the end of the trial matched predicted equilibrium concentrations. In contrast, measured PO4-P concentrations were approximately 40 – 50% lower than predicted regardless of diets, suggesting that PO4-P was likely utilized by RAS microbes. We therefore speculate whether lowering dietary P levels could become an effective tool for microbial management in RAS.

Water level control of small-scale recirculating aquaculture system with protein skimmer using fuzzy logic controller

The recirculating aquaculture system (RAS) is a land-based aquaculture facility, either open-air or indoors, that minimizes water consumption by filtering, adapting, and reusing water. Solid organic matter from fish waste and food waste directly becomes waste that needs to be eliminated because it is a source of increasing total ammonia nitrogen (TAN), total suspended solids (TSS), total dissolved solids (TDS), and also has an impact on reducing dissolved oxygen (DO). RAS requires a water level control system so the fish tank does not experience water shortages or floods, disrupting the aquatic aquaculture ecosystem. In this study, small-scale RAS is modeled using a 3-coupled tanks system approach with a tank configuration that follows the most straightforward RAS water recirculation process (fish tank, mechanic filter, biofilter). Clean water from the reservoir flows into the fish tank through a protein skimmer. This study applies the fuzzy logic controller (FLC) to control the water level in the protein skimmer and biofilter tanks by controlling the position of several valves where the placement positions of the valves have been determined according to system requirements. The study results show that the tuned single-input FLC has the best average output response characteristics with t&lt;sub&gt;s&lt;/sub&gt;=50, h&lt;sub&gt;1ss&lt;/sub&gt;=49.98, e&lt;sub&gt;ss&lt;/sub&gt;=0.02 in protein skimmer and t&lt;sub&gt;s&lt;/sub&gt;=4700, h&lt;sub&gt;1ss&lt;/sub&gt;=39.75, e&lt;sub&gt;ss&lt;/sub&gt;=0.25 in the tank system.

Comparison of the gill and gut microbiomes of common carp (Cyprinus carpio) and zebrafish (Danio rerio) and their RAS environment

Recirculating aquaculture systems (RAS) are increasingly being used to grow fish, as intensive water reuse reduces water consumption and environmental impact. RAS use biofilters containing nitrogen-cycling microorganisms that remove ammonia from the aquaculture water. Knowledge of how RAS microbial communities relate to the fish-associated microbiome is limited, as is knowledge of fish-associated microbiota in general. Recently, nitrogen-cycling bacteria have been discovered in zebrafish and carp gills and shown to detoxify ammonia in a manner similar to the RAS biofilter. Here, we compared RAS water and biofilter microbiomes with fish-associated gut and gill microbial communities in laboratory RAS housing either zebrafish (Danio rerio) or common carp (Cyprinus carpio) using 16S rRNA gene amplicon sequencing. The phylogeny of ammonia-oxidizing bacteria in the gills and the RAS environment was investigated in more detail by phylogenetic analysis of the ammonia monooxygenase subunit A (amoA). The location from which the microbiome was sampled (RAS compartments and gills or gut) had a stronger effect on community composition than the fish species, but species-specific differences were also observed. We found that carp- and zebrafish-associated microbiomes were highly distinct from their respective RAS microbiomes, characterized by lower overall diversity and a small core microbiome consisting of taxa specifically adapted to the respective organ. The gill microbiome was also defined by a high proportion of unique taxa. Finally, we found that amoA sequences from the gills were distinct from those from the RAS biofilter and water. Our results showed that the gut and gill microbiomes of carp and zebrafish share a common and species-specific core microbiome that is distinct from the microbially-rich RAS environment.

Optimal Dietary Protein/Energy Ratio and Phosphorus Level on Water Quality and Output for a Hybrid Grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀) Recirculating Aquaculture System

In a recirculating aquaculture system (RAS), feed is critical to the growth of fish and is the main source of nutrient pollutants in aquaculture water. An eight-week feeding trial was conducted to investigate the role of feed on the growth efficiency of hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀) and water quality in a RAS. Five commercial feeds with different respective dietary protein/energy (P/E) ratios and available phosphorus levels were selected (LNLP, 31.97 g/MJ, 0.96%; LNMP, 32.11 g/MJ, 1.54%; MNLP, 36.26 g/MJ, 0.98%; MNMP, 36.53 g/MJ, 1.58%; and HNP, 41.54 g/MJ, 1.97%). The results showed that HNP had the highest growth efficiency and MNLP provided the best economic benefit. The trend in water quality within 6 h after feeding was similar among the five groups. The relative concentrations of ammonia nitrogen, total nitrogen, active phosphate, and total phosphorus reached a maximum 2 h after feeding, and the relative concentration of nitrite reached a maximum 1 h after feeding. The high P/E ratio feed increased the concentrations of total ammonia nitrogen and nitrite nitrogen. The total ammonia nitrogen concentration in HNP was much higher than those in the other treatments. The dietary P/E ratio had no significant effect on total nitrogen concentration. High dietary phosphorus levels increased the total phosphorus concentration in the water, but no significant effect on the active phosphate concentration was observed. Considering the growth efficiency, economic benefit, and water quality, it can be concluded that MNLP is the most suitable feed for RAS breeding hybrid grouper. The results of this study supplement the gap on the effects of feed on RAS water quality and provide data support for the sustainable development of RAS industry.

Toward Feeds for Circular Multitrophic Food Production Systems: Holistically Evaluating Growth Performance and Nutrient Excretion of African Catfish Fed Fish Meal-Free Diets in Comparison to Nile Tilapia

In aquaponics and circular multitrophic food production systems, dietary protein source, as well as fish species choice, particularly in cases of different nutritional physiology, could be factors affecting excreted nutrient profiles. Accordingly, growth performance, dissolved nutrient accumulation and feces nutrient profiles were evaluated for African catfish (Clarias gariepinus) reared in recirculating aquaculture systems (RAS) and fed single protein source diets based on black soldier fly larvae meal (BSF), poultry by-product meal (PM), poultry blood meal (PBM) and fish meal (FM) and the results were compared to previous findings for Nile tilapia (Oreochromis niloticus). All diets resulted in significantly different growth performances of African catfish, with FM producing the best growth performance, followed by PM, BSF and PBM. PM resulted in the highest soluble reactive phosphorus concentrations (SRP) in the RAS water; whereas, BSF resulted in the highest K, Mg and Cu concentrations. The highest feces nutrient density was recorded for PBM; whereas, FM and PM yielded the lowest feces nutrient density. Comparing African catfish to Nile tilapia revealed that the former showed significantly better growth performance with FM and PM, however, significantly weaker performance with BSF. Although dissolved K accumulation was similar between species across diets, significant differences were recorded for total inorganic nitrogen and SRP production per unit of feed for individual diets. Despite similar feces nutrient profiles, African catfish produce significantly less feces dry matter per unit of feed for each diet compared to Nile tilapia. Findings are discussed regarding their implications for aquafeed development in the context of circular multitrophic food production systems.

Efficacy of H2O2 on the removal kinetics of H2S in saltwater aquaculture systems, and the role of O2 and NO3−

The occurrence of hydrogen sulfide (H2S) represents a challenge for recirculating aquaculture systems (RAS) under saline conditions. Even low concentrations of the toxic gas can result in sudden mass mortalities of fish, leading to large economic losses. There is an urgent need for efficient strategies to remove H2S, which can be applied effectively with a short response time, to prevent the risk of H2S-induced casualties. This study examines the kinetics of the two common oxidants applied to rearing water in a RAS facility; oxygen (O2) and hydrogen peroxide (H2O2) and evaluates their efficiency and applicability for the removal of H2S in an industrial RAS. Furthermore, we tested whether nitrate (NO3−) can be an oxygen donor in the chemical oxidation of H2S. The baseline oxidation rates of H2S by O2 were determined in air-equilibrated seawater (SW) and RAS water (RASW). The feasibility of using H2O2 as a practical treatment was evaluated by testing increasing H2O2 to H2S ratios in SW. In addition, RASW dilutions that yielded different concentrations of NO3− and total chemical oxygen demand (TCOD) were tested to identify their effects on H2S removal. The half-lives (t½) of H2S, derived from O2 oxidation rates, were considerably shorter in SW (118.5 ± 28.6 min) compared to RASW (168.0 ± 18.7 min). The addition of a 1:1 mole ratio of H2O2 to H2S, significantly increased the removal rate and decreased the half-life (t½) of H2S in SW to 29.5 ± 6.6 min. Further increasing H2O2:H2S ratios to 2:1 and 4:1, reduced t½ to 21.7 ± 5.2 and 17.4 ± 6.1 min, respectively. Similarly, a dosage of H2O2 at a ratio of 1:1 in RAS water resulted in a considerably shorter t½ of 86.1 ± 10.1 min. The influence of organic matter on the required H2O2 dose was demonstrated by the t½, which were reduced by 49% in RAS water and 75% in SW. NO3− was not found to be involved in the chemical removal of H2S. The results provide an improved understanding of the influence of RAS water chemistry and quality on H2S kinetics and the direct applicability of the kinetics for treating acute H2S levels in RAS to avoid mass mortalities. In conclusion, the addition of H2O2 is an efficient water treatment technology for H2S removal, and by adjusting H2O2 dosages accordingly to the concentrations of H2S and specific systems water parameters, a t½ <30 min can be achieved. Thus the technology is applicable in an industrial RAS, as a treatment process for acute levels of the hazardous gas H2S that is easily implemented, and safe for the fish.

Intensified fish farming. Performance of electrochemical remediation of marine RAS waters

Aquaculture has been the fastest growing agricultural sector in the past few decades and currently supplies about half of the fish market. A range of environmental and management concerns including limited land and water availability have led to intensifying fish production by recirculating aquaculture systems (RAS). Fish's diet contains 30–60 % protein and about 4–10 % nitrogen (N). As fish assimilate only 20–30 % of the feed to produce body mass, the unassimilated N is released in the form of toxic ammonium that deteriorates water quality and compels its degradation. Widely extended biological nitrification is not efficient in the removal of nitrites nor other chemicals and pharmaceuticals used during fish culture. Electrochemical oxidation, a less developed alternative, reports several advantages such as, i) simultaneous degradation of ammonia‑nitrogen (TAN) and water disinfection in the same step with considerable simplification of the whole process, ii) easy adaptability to different production scales and periods of fish growth, and iii) no generation of harmful by-products and no use of chemicals, among others. Besides, in the case of marine aquaculture, the technology benefits from the high conductivity of seawater; thus, electrochemical oxidation is positioned in a very good place to satisfy the water treatment needs of the increasing production rate of marine aquaculture fish. Here, we report the analysis of the performance of a RAS demonstration plant aimed at farming gilthead sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax) and provided with electrochemical remediation of culture water. The performance of the plant, with 20 m3 of seawater operating at a recirculation rate of 0.9–1.4 h−1, has been analysed in terms of TAN removal, water disinfection, make-up water intake and energy consumption and compared to data of conventional RAS provided with biofilters. The benefits and advantages of the innovative electrochemical remediation of RAS water are highlighted.

Exposing Atlantic Salmon Post-Smolts to Fluctuating Sublethal Nitrite Concentrations in a Commercial Recirculating Aquaculture System (RAS) May Have Negative Consequences

Salmon farmers are interested in extending the time post-smolts are reared in recirculating aquaculture systems (RAS). However, there is a lack of knowledge regarding optimal water quality for post-smolts in RAS, and regarding potential consequences of long term exposure to different toxic compounds, such as nitrite, in the RAS water. To address this issue, we conducted a case study at a Faroese Atlantic salmon farm, that rears large post-smolts in fresh water RAS for 22 months before sea transfer, with no additional chloride salt, a known treatment for nitrite toxicity. The aim was to document the potential effects of long-term exposure of fluctuating sub-lethal nitrite concentrations in fresh water RAS on blood physiology of large post-smolts. The study was conducted over six weeks, at the end of the RAS production cycle. Our case study shows that after ~22 months in RAS with no additional chloride, the fish had accumulated a plasma nitrite concentration 8 to 16 times higher than the ambient water. Our results indicate that the accumulation may have resulted in extracellular hyperkaliemia, since there was a positive correlation between plasma nitrite and potassium levels (p=0.00095), with potassium levels almost twice as high than previously reported for Atlantic salmon. This could indicate that Atlantic salmon health is challenged due to prolonged sub-lethal nitrite exposure in fresh water RAS. Further research related to long-term nitrite exposure in RAS is needed to asses the potential negative impact, in order to optimize welfare and growth performance during production of Atlantic salmon post-smolts.

Phosphorus recovery from recirculating aquaculture systems: Adsorption kinetics and mechanism

Recirculating aquaculture systems (RAS) have grown in popularity in recent years due to their potential to provide a high-quality protein source in a contained environment. With increased production comes the need for RAS wastewater treatment to remove waste products such as phosphorus, which can harm other aquatic life in the area by causing algae blooms. Additionally, there is potential to harvest the wasted phosphorus for use as a fertilizer and to combat nutrient scarcity. This study investigates a phosphorus adsorbent, RhizoSorb® (base material of aluminum oxide), for its applicability in RAS use under fresh water, simulated seawater, and real RAS water conditions. Film diffusion and intraparticle diffusion were both rate-controlling steps in the adsorption process, and the good fit of the Elovich model indicated that chemisorption is likely the dominating adsorption mechanism. Phosphate removal decreased with increasing salinity in all tests, which can be attributed to both anion competition and increasing pH. The Freundlich isotherm fit the equilibrium data better than the Langmuir isotherm, showing that adsorption is a multi-layer process and that the adsorbent is highly heterogeneous. The Clark model was better suited than the Thomas model for predicting the performance of the adsorbent in a flow-through system. The adsorbent showed high removal efficiencies and good selectivity for phosphate in all tests.

Impact of ozone treatment on dissolved organic matter in land-based recirculating aquaculture systems studied by Fourier transform ion cyclotron resonance mass spectrometry

In land-based recirculating aquaculture systems (RAS), the accumulation of dissolved organic matter (DOM) can have detrimental effects on water quality impacting the system performance, microbial community, and consequently fish health and welfare. Ozone is used in the RAS water treatment process to improve water quality and remove DOM. However, little is known about the molecular composition of DOM in RAS and its transformation when exposed to ozone. In this study, we performed a detailed molecular characterization of DOM in RAS and explored its transformation induced by ozonation of RAS waters. Ultra-high resolution (UHR) Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) was used to characterize the DOM matrix of RAS waters (pump-sump and tanks) and to evaluate its transformation by ozonation. The analysis of DOM extracted from makeup water and feed samples allowed for the determination of DOM sources in RAS prior to ozonation. The CHO and unsaturated group of compounds were the most abundant class found in water samples. On the contrary, the DOM from feed samples was unique and consisted mainly of CHO, CHON and unsaturated group of compounds. After the ozonation of RAS waters, humic-like and unsaturated compounds [positive oxygen subtracted double bond equivalent per carbon (DBE–O)/C)] were decomposed, particularly the CHO-DOM that contained fewer -CH2- features. Fulvic-like compounds and several hundred saturated compounds [negative (DBE–O)/C)] were formed post ozonation, particularly the CHON and CHONS group of compounds that were associated with fish diets, makeup waters and transformation products from the ozonation of the RAS waters. This study showed that the high accuracy of the ultra-high resolution FTICR MS can be applied to characterize and monitor the changes of DOM at a molecular level in RAS waters. To our knowledge, this is the first study where FTICR MS was incorporated for the characterization of DOM and its sources in RAS.

Changes in rearing water microbiomes in RAS induced by membrane filtration alters the hindgut microbiomes of Atlantic salmon (Salmo salar) parr

An important step in water treatment in recirculating aquaculture systems (RAS) is the removal of particulate organic matter. While commonly applied treatment technologies remove most of the particles, fine particulate organic matter typically accumulate in RAS, and serve as substrate for heterotrophic bacteria in the system. Membrane ultrafiltration has been shown to reduce the concentrations of total organic carbon and to counteract blooms of opportunistic bacteria in periods with high organic loading. The potential influence of such opportunistic bacteria on the fish microbiomes and fish welfare is poorly understood. In this study, we investigated the effects of high organic loading on the water and hindgut microbiomes in RAS with Atlantic salmon parr. Two pilot-scale RAS, one with membrane ultrafiltration of a 10–15% side-stream (mRAS), and one without (cRAS), were operated in periods with low and high organic loadings, and the water and hindgut microbiomes were characterized by Illumina sequencing of 16S rDNA amplicons. At the end of the experiment, after a period of high organic loading, two OTUs representing Mycobacterium and Spartobacteria dominated the water microbiomes in cRAS, with relative abundances as high as 24 and 17%, respectively. In the mRAS water they were rare (relative abundances of 1.9 and 0.1%). The membrane ultrafiltration stabilized the water microbiota and efficiently prevented the growth of these bacterial populations in RAS. The parr hindgut microbiomes were generally highly distinct from the RAS water microbiomes and were dominated by the bacterial phyla Bacilli and Firmicutes. Furthermore, an OTU probably representing Carnobacterium inhibens, originally isolated from the gut of adult salmon, was highly abundant in the gut microbiomes, with relative abundances reaching over 30%. However, for the cRAS fish after a period with high organic loading, the relative abundance of this Carnobacterium OTU was reduced, and an increased similarity between the gut and the water microbiomes was observed. The Mycobacterium and Spartobacteria OTUs that were abundant in the cRAS water were also abundant in the cRAS hindgut samples. This indicates a direct influence of the water microbiomes on the gut microbiomes, and that high organic loading in the cRAS resulted in growth of opportunistic bacteria that were able to colonize the fish gut. The present study shows that membrane filtration is an applicable strategy to counteract growth of opportunistic bacteria in RAS and to promote positive fish – microbe interactions.

Design of an Intelligent Variable-Flow Recirculating Aquaculture System Based on Machine Learning Methods

A recirculating aquaculture system (RAS) can reduce water and land requirements for intensive aquaculture production. However, a traditional RAS uses a fixed circulation flow rate for water treatment. In general, the water in an RAS is highly turbid only when the animals are fed and when they excrete. Therefore, RAS water quality regulation technology based on process control is proposed in this paper. The intelligent variable-flow RAS was designed based on the circulating pump-drum filter linkage working model. Machine learning methods were introduced to develop the intelligent regulation model to maintain a clean and stable water environment. Results showed that the long short-term memory network performed with the highest accuracy (training set 100%, test set 96.84%) and F1-score (training 100%, test 93.83%) among artificial neural networks. Optimization methods including grid search, cuckoo search, linear squares, and gene algorithm were proposed to improve the classification ability of support vector machine models. Results showed that all support vector machine models passed cross-validation and could meet accuracy standards. In summary, the gene algorithm support vector machine model (accuracy: training 100%, test 98.95%; F1-score: training 100%, test 99.17%) is suitable as an optimal variable-flow regulation model for an intelligent variable-flow RAS.

Effect of different C/N ratios and hydraulic retention times on denitrification in saline, recirculating aquaculture system effluents

Data on operation and performance of cost-effective solutions for end-of-pipe removal of nitrate from land-based saltwater recirculating aquaculture systems (RAS) are scarce but increasingly requested by the aquaculture industry. This study investigated the performance of a (semi)commercial-scale fixed-bed denitrification unit using single sludge for treating effluent from a commercial, saltwater RAS used for production of Atlantic salmon (Salmo salar). A fixed-bed denitrification reactor was fed continuously with 3-days hydrolyzed sludge from the commercial RAS, and was operated at different hydraulic retention times (HRTs; 1.82, 3.64, 5.46, or 7.28 h) or influent C/N ratios (3, 5, 7, or 10). Twenty-four h pooled samples were collected from the inflowing RAS water and the hydrolyzed sludge as well as from the denitrification reactor outlet, and samples were analyzed for nutrients and organic matter content.Nitrate removal rates increased consistently with decreasing HRT (from 64.3 ± 5.2–162.7 ± 22.0 g NO3-N/m3/d within the HRTs tested) at non-limiting C/N ratios, while nitrate removal efficiencies decreased (from 99.6 ± 0.3–58.2 ± 8.9 %). With increasing influent C/N ratios at constant HRT (3.64 h), nitrate removal rates increased until the removal efficiency was close to 100 % and nitrate concentration in the denitrification reactor became rate-limiting. A maximum nitrate removal rate of 162.7 ± 2.0 g NO3-N/m3/d was achieved at a HRT of 1.82 h and an influent C/N of 6.6 ± 0.5, while the most efficient use of hydrolyzed sludge (0.19 ± 0.02 g NO3-N removed/g sCOD supplied) was obtained with a HRT of 3.64 h and a C/N ratio of 2.9. Removal rates of organic matter significantly and consistently increased with decreasing HRT and increasing C/N ratio. In addition, reducing HRT and increasing C/N ratios significantly improved removal of total phosphorus (TP) and PO4-P.In conclusion, optimal management of the operating parameters (HRT and C/N ratio) in a single-sludge denitrification process can significantly reduce the discharge of nitrogen, organic matter, and phosphorous from land-based saltwater RAS and thus contribute to increased sustainability.

Environmental and energy requirements for different production biomass of Nile tilapia (Oreochromis niloticus) in recirculating aquaculture systems (RAS) in Kenya

Recirculating Aquaculture Systems (RAS) offers a better option to increase aquaculture production with limited land and water resources while minimizing water pollution. The biggest challenges in RAS is to maintain favorable water quality for the fish to thrive. The practice of RAS in Kenya is minimal with the improper matching of RAS components and production densities for the existing few and in most cases, leading to a system failure. This study aimed at evaluating environmental and energy requirements for different production biomass of Nile tilapia in a RAS established in a controlled environment. Both production densities and water flow rates were varied while at the same time, the water quality parameters (dissolved oxygen, ammonia, pH, electrical conductivity and temperature) were monitored. The energy consumed for pumping and aeration was also monitored. Tilapia stocking biomass varied from 2.3 kg/m3 to 10 kg/m3 while flow rate was varied from 2.0 L/min and increased at intervals of 1.0 L/min to the maximum attainable flow rate of 10.0 L/min. Crushed pumice rock packed in a 1000 L tank and equipped with a bell siphon was used as the biofilter. Ammonia removal reduced with increasing flow rate with removal rates of 75% at 2.0–3.0 L/min to 9% at 8.0–10.0 L/min. The water pH increased with increasing flow rate with R2 ranging from 0.4 to 0.9. Electrical conductivity increased with flow rate from 112 mg/L to as high as 209 mg/L. Dissolved oxygen increased with flow rate and ranged from 1.0 mg/L to 7.0 mg/L. Energy consumption increased with flow rate and raged from 0.4 kW h at 2.3 kg/m3 stocking biomass to 2.4 kW h at 10 kg/m3 stocking biomass. High stocking densities and flow rates resulted to lower purification efficiencies and higher power consumptions than low stocking densities and flow rates. In order to maintain good RAS water quality, increased production and profits among farmers using RAS in Kenya, there is need to give proper advice on the right combination of stocking biomass, power, and water flow rate. In addition, similar studies should be carried out for other common fish species such as the African catfish.

Co-cultivation of microalgae in aquaculture water: Interactions, growth and nutrient removal efficiency at laboratory- and pilot-scale

Microalgae biotechnology is of increasing importance and a central application concerns the treatment of wastewater. Here, its implementation in a recirculating aquaculture system (RAS) to lower the discharge of wastewater is studied. To better cope with external variations in culture conditions, a co-cultivation of two species of microalgae, Chlorella vulgaris and Tetradesmus obliquus, was used to obtain a more reliable and robust culture and was compared to monocultures. This approach was tested using RAS water both under sterile and non-sterile conditions at laboratory scale and then compared to a co-culture at pilot-scale in an open thin-layer photobioreactor. Performance of cultures was tested in terms of microalgae growth and nutrient removal efficiency. Furthermore, to better understand the interaction between environmental variables and each microalgae species, their relative frequencies in co-cultures as well as the presence of protozoa and bacteria were monitored. All growth experiments were carried out successfully and, unlike in a previous study, no crashes were observed. However, shifts in species frequency in co-cultures indicated that the two species were differentially affected by cultivation conditions. Despite nutrient limitation, the pilot-scale cultivation had a high productivity (13.3 g m−2 d−1) and final dry weight (11.1 g l−1) after 29 days and demonstrated its suitability for RAS water treatment.

Effects of reduced organic matter loading through membrane filtration on the microbial community dynamics in recirculating aquaculture systems (RAS) with Atlantic salmon parr (Salmo salar)

A key challenge in recirculating aquaculture systems (RAS) is the accumulation of particulate organic matter, especially the fine and colloidal fraction due to low removal efficiency of today's technology. The supply of organic matter is typically the limiting resource determining the carrying capacity (CC) of heterotrophic bacteria in the system. An appropriate and stable CC is proposed as a strategy for an optimal microbial environment in RAS with less blooms of opportunistic bacteria and more stable community dynamics. In this study, we investigated the effects of including a membrane for ultrafiltration in the RAS water treatment loop (treating 10–15% of the total water flow) to reduce the amount of fine and colloidal organic matter. Atlantic salmon parr (Salmo salar) were reared in two pilot-scale RAS (mRAS: membrane, cRAS: conventional). To evaluate the bacterial dynamics with and without membrane filtration at different organic loadings, the water exchange rates of the systems were manipulated equally to create periods with high and low loading of organic matter. The results showed that in the mRAS water, the level of organic matter was more stable throughout the experiment for the changing organic matter loadings. As a consequence, water in mRAS had higher microbial diversity, lower and shorter bacterial blooms and generally lower bacterial densities than in cRAS. All variables indicate a better microbial environment in the water of the system with membrane filtration. Also, the physicochemical water quality was better in mRAS in terms of lower turbidity and particulate organic matter (POC), and slightly lower concentrations of total ammonia nitrogen (TAN). The composition of the microbial communities was significantly different between the two systems, and temporal variations in the community dynamics were observed in both systems during the periods with different organic loadings. At high organic loading, the genus Mycobacterium had high relative abundance in the cRAS water (up to 0.25) compared to mRAS (0.01–0.03). The fish in mRAS were significantly bigger (14%) than fish in cRAS at the end of the experiment, however it is hard to conclude whether the better growth in mRAS was due to higher temperatures (caused by membrane operation) or better water quality, as it was probably a combination of both. We can conclude that membrane filtration gave more stable and better physicochemical and microbial water quality, which will reduce the probability for microbially related accidents in RAS.