Marine Recirculating Aquaculture System Research Articles

Occurrence, distribution and sources of microplastics in typical marine recirculating aquaculture system (RAS) in China: The critical role of RAS operating time and microfilter

Industrial mariculture, a vital means of providing high quality protein to humans, is a potential source of microplastics (MPs) which have recently received increasing attention. This study investigated the occurrence and distribution of microplastics in feed, source water and recirculating aquaculture system (RAS) with long & short operating times as well as in fish from typical industrial mariculture farms in China. Results showed that microplastics occurred in all samples with the average concentration of 3.53 ± 1.39 particles/g, 0.70 ± 0.17 particles/L, 1.53 ± 0.21 particles/L and 2.21 ± 0.62 particles/individual for feed, source water, RAS and fish, respectively. Microplastics were mainly fiber in shape, blue in color and 20–500 μm in size. Compared with short operated RAS, long operating time led to higher microplastic concentration in RAS, especially that of microplastic in 20–500 μm, granular and blue. Regardless of short or long operating time, microplastics in RAS mainly gathered in culture tank, tank before microfilter and fixed-bed biological filter, and the microfilter removed efficiently the microplastic with the shape of film, granule, fragment as well as those with size > 1000 μm. As for the polymer types, polyamide (PA, 71.9 %) and polyethylene terephthalate (PET, 65.7 %) dominated in feed and source water, respectively, which may be the reason for the high proportion of PA (38.8 % and 26.4 %) and PET (31.8 % and 30.2 %) in RAS and fish. In addition, polypropylene (PP) was also detected in RAS (18.7 %) and fish (22.6 %), indicating that other plastic facilities such as PP brush carrier also made a contribution. Positive matrix factorization (PMF) model revealed three sources of MP in RAS, namely plastic facilities, industrial sewage and plastic packaging products. Our results provided a theoretical basis for the management of MP in RAS.

RETRACTED: Enhanced biofiltration coupled with ultrafiltration process in marine recirculating aquaculture system: Fast start-up of nitrification and long-term performance

In marine recirculating aquaculture systems (RAS), the biological nitrification system usually suffers from long time start-up and proliferation of potential pathogenic bacteria under the high salinity stress. This study aims to develop a strategy for fast start- up and long-term biosecurity of marine RAS by using a combined biofiltration system (ceramic rings (CR) - granular activated carbon (GAC) - ultrafiltration (UF) process) in an actual marine RAS for selenotoca multifasciata. The ammonia nitrogen and dissolved organic matter (DOM) removal performance, membrane fouling characteristics and microbial communities during long-term operation (200 days) were investigated. The CR-GAC-UF process minimized the start-up time of the nitrification system to 14 days under low ammonia concentrations (below 2 mg/L). Long-term operation of the marine RAS with ammonia and nitrite nitrogen stabilized below 0.1 mg/L with a daily ammonia nitrogen removal rate of 91.7 %. The UF membranes were operated at fluxes at 15 LHM and the protein/polysaccharide ratios of soluble microbial products (SMP) and extracellular polymeric substances (EPS) on the membranes were stable below 0.5 with low fouling levels. A number of bacterial genera that have the potential to degrade protein and humic-like substances were recognized, which was related to the long-term DOM removal. Candidatus_Nitrosopumilus, Nitrosomonas_sp. and Nitrospira sp. ENR4 were detected as the core nitrifiers in CR-GAC-UF process, which were closely related to the amoABC, hao and nxrA genes (p < 0.05). With the multi-stage barrier of CR-GAC-UF, the heterotrophic plate count in therecirculating water was kept at 3 ± 1 CFU/mL and the relative abundance of potential pathogens declined gradually below 0.81 %. Taken together, this study provides a theoretical basis for the wider application of marine RAS.

Characterization of simultaneous ammonium and nitrate removal and microbial communities in airlift reactor using 3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) as carbon source and biofilm carrier

As a novel trend, solid carbon sources are applied to act as electron donors and biofilm carrier in biological denitrification process. In this study, simultaneous nitrate and ammonium removal process in an airlift sequencing batch reactor using 3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) as carbon source and biofilm carrier under intermittent aeration conditions was established to treat effluent of synthetic marine recirculating aquaculture system. The results showed that maximum nitrate and ammonia nitrogen removal rates of 0.45 and 0.09 kg m−3 d−1 were achieved. No significant nitrite accumulation was found during 200-day operation, while effluent dissolved organic carbon accumulation and particle size reduction significantly increased. Microbial community analysis and batch tests illuminate that the generated sludge and attached biofilm played important roles in nitrogen removal. This study demonstrates the potential mechanism for the nitrogen removal process mediated by 3-hydroxybutyrate-co-3-hydroxyvalerate and provide a new idea for the alternative solutions of solid carbon sources.

Comparision of nitrogen removal characteristic and microbial community in freshwater and marine recirculating aquaculture systems

Recirculating aquaculture system (RAS) has a good prospect in aquaculture, but its nitrogen removal characteristics and microbial community changes in freshwater and marine water remain unclear. In this study, six RAS were designed and divided into freshwater group and marine water group with salinity of 0‰ and 32‰, respectively, and operated for 54 days to test changes in nitrogen (NH4+-N, NO2--N, NO3--N), extracellular polymeric substances and microbial communities. The results showed that ammonia nitrogen was rapidly reduced and almost converted to nitrate nitrogen in the freshwater RAS but to nitrite nitrogen in marine RAS. Compared with freshwater RAS, marine RAS had lower tightly bound extracellular polymeric substances and worse stability and settleability condition. 16S rRNA amplicon sequencing reflected significantly lower bacterial diversity and richness in marine RAS. Microbial community structure at phylum level showed lower relative abundance of Proteobacteria, Actinobacteria, Firmicutes, Nitrospirae, but higher abundance of Bacteroidetes under a salinity of 32‰. High salinity decreased the abundance of funtional genera (Nitrosospira, Nitrospira, Pseudomonas, Rhodococcus, Comamonas, Acidovorax, f_Comamonadaceae), which may account for nitrite accumulation and low nitrogen removal capacity in marine RAS. These findings could provide theoretical and practical basis for improving the start-up speed of high-salinity nitrification biofilm.

Candidatus Scalindua, a Biological Solution to Treat Saline Recirculating Aquaculture System Wastewater

Recirculating aquaculture systems (RAS) are promising candidates for the sustainable development of the aquaculture industry. A current limitation of RAS is the production and potential accumulation of nitrogenous wastes, ammonium (NH4+), nitrite (NO2−) and nitrate (NO3−), which could affect fish health and welfare. In a previous experiment, we have demonstrated that the marine anammox bacteria Candidatus Scalindua was a promising candidate to treat the wastewater (WW) of marine, cold-water RAS. However, the activity of the bacteria was negatively impacted after a direct exposure to RAS WW. In the current study, we have further investigated the potential of Ca. Scalindua to treat marine RAS WW in a three-phase experiment. In the first phase (control, 83 days), Ca. Scalindua was fed a synthetic feed, enriched in NH4+, NO2− and trace element (TE) mix. Removal rates of 98.9% and 99.6% for NH4+ and NO2−, respectively, were achieved. In the second phase (116 days), we gradually increased the exposure of Ca. Scalindua to nitrogen-enriched RAS WW over a period of about 80 days. In the last phase (79 days), we investigated the needs of TE supplementation for the Ca. Scalindua after they were fully acclimated to 100% RAS WW. Our results show that the gradual exposure of Ca. Scalindua resulted in a successful acclimation to 100% RAS WW, with maintained high removal rates of both NH4+ and NO2− throughout the experiment. Despite a slight decrease in relative abundance (from 21.4% to 16.7%), Ca. Scalindua remained the dominant species in the granules throughout the whole experiment. We conclude that Ca. Scalindua can be successfully used to treat marine RAS WW, without the addition of TE, once given enough time to acclimate to its new substrate. Future studies need to determine the specific needs for optimal RAS WW treatment by Ca. Scalindua at pilot scale.

Abundance and Diversity of Nitrifying Microorganisms in Marine Recirculating Aquaculture Systems

Recirculating aquaculture systems (RAS) are important for water quality management in aquaculture facilities, and can help resume water consumption. However, information about the community structure of the micro-ecosystem existing in biofilters, especially the participation of the known nitrifying groups (i.e., AOA, AOB, NOB, and comammox Nitrospira), remains to be fully clarified. In this research, we compared the community structures in three RAS systems operated at different temperatures in a marine aquarium, through both amoA-targeted qPCR assay and 16S rRNA-targeted next-generation sequencing. As result, AOA was the primary nitrifier in the biofilters and was typically abundant and diverse in high-temperature samples (ca. 25 °C). NOB’s relative abundance patterns were numerically similar to that of AOA, suggesting a cooperation relationship between AOA and NOB in the marine RAS system. AOB was at a comparable level with AOA in medium-temperature samples (ca. 19 °C), while their abundance sharply decreased in high-temperature samples. The number of observed OTUs of AOA in high-temperature samples was 1.9 and 1.5 times as much as that detected in low (ca. 10 °C) and medium temperature samples respectively, suggesting a much more diverse and predominant occurrence of AOA at high temperatures. Comammox Nitrospira was only detected at a low level in the biofilter samples, suggesting a negligible contribution to the nitrification process in such ammonia-limited, saline biofilms. Although comammox Nitrospira cannot be detected by 16S rRNA-based analysis, the high diversity and abundance of NOB that were detected in high-temperature samples indicated the prospective possibility of the occurrence of complete ammonia oxidation at high temperatures.

Dethiosulfovibrio faecalis sp. nov., a novel proteolytic, non-sulphur-reducing bacterium isolated from a marine aquaculture solid waste bioreactor.

A new Dethiosulfovibrio strain, designated F2BT, was isolated from an anaerobic digester for treating solid waste from a marine recirculating aquaculture system. The motile, Gram-negative, non-spore-forming curved rods were 2-7 µm long and 1 µm in diameter. Growth occurred at temperatures ranging from 20 to 40 °C with a maximum rate of growth at 30 °C. The pH range for growth was pH 6.0-8.0, with a maximum rate of growth at pH 7.5. This isolate was halotolerant growing in NaCl concentrations ranging from 0 to 1.6 M with a maximum rate of growth at 0.4 M. Similarly to the five described Dethiosulfovibrio species, this obligate anaerobe isolate was fermentative, capable of utilizing peptides, amino acids and some organic acids for growth, but unlike described strains in the genus did not reduce thiosulphate or elemental sulphur to hydrogen sulphide during fermentation of organic substrates. The G+C content of 55 mol% is similar to the described Dethiosulfovibrio species. The average nucleotide identity analysis between whole genome sequences showed less than 93.15% sequence similarity between strain F2BT and the five other described Dethiosulfovibrio species. Differences in the physiological and phylogenetic characteristics between the new strain and other Dethiosulfovibrio specied indicate that F2BT represents a novel species of this genus and the epithet Dethiosulfovibrio faecalis sp. nov. is proposed. The type strain is F2BT (=DSM 112078T=KCTC25378T).

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.

Dynamics of Water and Biofilm Bacterial Community Composition in a Mediterranean Recirculation Aquaculture System

Recirculation technology has been emerging in the marine aquaculture industry. The microbiome developed in recirculation aquaculture systems (RASs) is an important factor for the optimal operation of these systems and fish welfare. In this study, the microbial community dynamics in the water column and the biofilms of a marine RAS with Mediterranean species of gilthead sea bream and sea bass were investigated, while physicochemical conditions were also monitored. Microbiological, culture, and non-culture analyses based on PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) fingerprints were performed on the water column and biofilm developed on stainless-steel surfaces. According to the obtained results, feed administration seemed to cause changes in pH and TAN, as well as drive changes in the bacterial abundance in the water column. Tested surfaces were colonized within 24 h and sessile cells were stabilized in terms of density within 6 days. DGGE fingerprints indicated the stability of the microbial community in water and a dynamic succession in the community of the biofilms. The fish pathogen Tenacibaculum discolor was found to colonize the biofilm and the water column. The main findings confirmed that RAS technology can be used as a control strategy for the stability of the water microbial community, that there is a dynamic succession of the dominant species in the biofilm communities, and that pathogenic bacteria can be dominant in the latter.

Microbial community dynamics in a commercial RAS for production of Atlantic salmon fry (Salmo salar)

Recirculating aquaculture systems (RAS) harbour complex microbial communities which can have an impact on the growth and development of the reared fish. This study aimed to improve our understanding of microbial community dynamics in a RAS involving three production batches of Atlantic salmon fry and parr during a period of 20 months. Water for analysis of microbiota was sampled at different positions in the RAS, and we also examined the effect of UV treatment on the water microbiota. Microbial communities were characterized by 16S rDNA amplicon sequencing of water samples taken directly upstream and downstream of the UV treatment unit and from three of the rearing tanks. In total 6 sampling events were made during a 20-month period. The study showed that: 1) Two of the production batches had a highly similar water microbiota despite disinfection of the system between the batches and rearing fish of different stages. In contrast, the first production batch showed a different water microbiota with variable composition through the system and over time. A more immature biofilter in the first batch may explain these differences. 2) The full-flow UV treatment directly upstream the rearing tanks had no observable effect on the community composition of the water microbiota in the different sampling positions in the RAS. This was likely a consequence of the low hydraulic retention time (HRT) (23 min) in rearing tanks, low bacterial regrowth in the fish tanks and community changes throughout the RAS loop. 3) The disinfection effect on viable bacterial densities in the water directly downstream of the UV treatment was around 89%, when the water was clear. Regrowth of bacteria following disinfection was low compared to those reported for marine RAS with UV disinfection and long HRT in fish tanks. The study shows that UV disinfection can be used to efficiently reduce bacterial density without compromising the microbial water quality in the fish tanks in RAS with low HRT.

Humic substances modulate fish bacterial communities in a marine recirculating aquaculture system

The stressful conditions of intensive aquaculture systems, combined with excessive antibiotic use, may have dysbiotic effects on aquaculture microbiomes and promote the spread of opportunistic pathogens. Here, we hypothesized that humic substances (HS), when added to the rearing water of a marine recirculating aquaculture system (RAS), will act as a chemical modulator of fish-associated bacterial communities and suppress potential pathogens. To test this, a 28-day RAS trial for juvenile European sea bass (Dicentrarchus labrax) was conducted with and without HS modulation. High-throughput sequencing of the 16S rRNA gene was used to evaluate role of HS in modulating fish bacterial assemblages (gut and skin mucus). In addition to this, biometric, digestive and oxidative stress parameters were measured to assess the impact of HS on fish performance. HS modulation was associated with an increased bacterial diversity and a distinct bacterial community composition in the fish skin mucus. HS modulation significantly increased the relative abundance of amplicon sequence variants (ASVs) related to the potentially beneficial Roseobacter clade and lowered the abundances of ASVs related to potential fish pathogens (e.g., Acinetobacter jonhsonii and Vibrio harveyi). In line with compositional differences, there was an enrichment of predictive KEGG categories related to antagonism and significantly lower abundances of KEGG categories related to pathogenesis and invasibility. In the fish gut, HS modulation was associated with a lower abundance of the orders Enterobacteriales and Bacillales. HS modulation also was associated with significant reductions in fish oxidative stress parameters and a significant increase in the activity of the digestive enzyme chymotrypsin. For the first time, our results demonstrate the potential of dissolved HS as a modulator of fish microbial communities.

Thermophilic bacteria combined with alkyl polyglucose pretreated mariculture solid wastes using as denitrification carbon source for marine recirculating aquaculture wastewater treatment

In marine recirculating aquaculture systems (RAS), efficient nitrogen removal is challenging due to the high NO3−-N concentration, low organic matters content, and high salinity. In this study, mariculture solid wastes (MSW) acidogenic liquid pretreated by thermophilic bacteria (TB) combined with alkyl polyglucose (APG) was first used as carbon source for denitrification to remove NO3−-N. TB + APG pretreatment could accelerate the hydrolysis of MSW, and the highest volatile fatty acids (VFAs) yield (40.3%) was obtained with TB + 0.2 g/g VSS APG pretreatment. MSW acidogenic liquid pretreated by TB + 0.2 g/g VSS APG was a reliable carbon source for denitrification, and the optimum COD/NO3−-N ratio (C/N) was 8 with no residue of NOx−-N. VFAs were more effectively utilized by denitrifiers than carbohydrate and protein. The high denitrification potential (PDN) and denitrification rate (VDN) indicated the higher denitrification ability at C/N of 8 using MSW acidogenic liquid as carbon source. The outcomes of this work could provide useful information for promoting technological innovation in marine RAS wastewater treatment.

Characterization of a novel marine aerobic denitrifier Vibrio spp. AD2 for efficient nitrate reduction without nitrite accumulation.

Aerobic denitrifiers have the potential to reduce nitrate in polluted water under aerobic conditions. A salt-tolerant aerobic denitrifier was newly isolated and identified as Vibrio spp. AD2 from a marine recirculating aquaculture system, in which denitrification performance was investigated via single-factor experiment, Box-Behnken experiment, and nitrogen balance analysis. Nitrate reductase genes were identified by polymerase chain reaction. Results showed that strain AD2 removed 98.9% of nitrate-nitrogen (NO3--N) with an initial concentration about 100 mg·L-1 in 48 h without nitrite-nitrogen (NO2--N) accumulation. Nitrogen balance indicated that approximately 17.5% of the initial NO3--N was utilized for bacteria synthesis themselves, 4.02% was converted to organic nitrogen, 39.8% was converted to nitrous oxide (N2O), and 31.1% was converted to nitrogen (N2). Response surface methodology experiment showed that the maximum removal of total nitrogen (TN) occurred under the condition of C/N ratio 11.5, shaking speed 127.9 rpm, and temperature 30.8 °C. Sequence amplification indicated that the denitrification genes, napA and nirS, were present in strain AD2. These results indicated that the strain AD2 has potential applications for removing NO3--N from high-salinity (3%) wastewater.

Wood and sulfur-based cyclic denitrification filters for treatment of saline wastewaters

This study investigated the performance and microbiome of cyclic denitrification filters (CDFs) for wood and sulfur heterotrophic-autotrophic denitrification (WSHAD) of saline wastewater. Wood-sulfur CDFs integrated into two pilot-scale marine recirculating aquaculture systems achieved high denitrification rates (103 ± 8.5 g N/(m3·d)). The combined use of pine wood and sulfur resulted in lower SO42− accumulation compared with prior saline wastewater denitrification studies with sulfur alone. Although fish tank water quality parameters, including ammonia, nitrite, nitrate and sulfide, were below the inhibitory levels for marine fish production, lower survival rates of Poecilia sphenops were observed compared with prior studies. Heterotrophic denitrification was the dominant removal mechanism during the early operational stages, while sulfur autotrophic denitrification increased as readily biodegradable organic carbon released from wood chips decreased over time. 16S rRNA-based analysis of the CDF microbiome revealed that Sulfurimonas, Thioalbus, Defluviimonas, and Ornatilinea as notable genera that contributed to denitrification performance.

Biofilters are potential hotspots for H2S production in brackish and marine water RAS

The production of hydrogen sulphide (H2S) is a major concern and challenge in marine land-based recirculating aquaculture systems (RAS). In order to gain a thorough understanding on H2S production dynamics in RAS, an anaerobic batch reactors experiment was performed, evaluating the production potential of total dissolved sulphide (TDS) and H2S and characterizing the microbial communities in different RAS environments (solid waste, biomedia+RAS-water, biomedia+seawater [SW], and RAS-water) during 33 days. The highest H2S concentrations were achieved in the reactors with biomedia+RAS-water, where biofilms growing on biomedia hosted a high abundance of sulphate reducing bacteria (SRB), which activated once anoxic conditions and carbon sources were introduced. The biofilms in the biomedia+SW reactors produced lower amounts of H2S than the biomedia+RAS-water reactors, due to the lower abundance of SRB. The solid waste produced H2S at low rate, the production being limited by the substrate competition between SRB and other microbial groups. No production of H2S was observed in the reactors with only RAS-water, due to the absence of SRB and suitable carbon sources. Based on measurements of production of H2S in this study, a moving bed biofilter submerged in brackish water (17 ppt) can potentially produce 139 g of H2S per biofilter m3, when the biomedia is not adequately mixed and oxygenated. Altogether, the results demonstrate that although SRB are found both in biofilters and solid waste, these two environments exhibit different H2S production dynamics. Biofilters produce large quantities of H2S under anoxic conditions, while solid waste deposited to the rearing tanks exhibits a lower but constant production of H2S. However, both of these H2S sources can be suppressed through fast removal of solids from rearing tanks and optimal operational conditions for biofilters.

Coagulation of phosphorous and organic matter from marine, land-based recirculating aquaculture system effluents

Saline effluents from marine land-based aquaculture production can neither be disposed in common municipal wastewater treatment plants, nor disposed as landfill. Furthermore, stricter environmental regulations require the reduction of phosphorous and organic matter levels from marine environment discharges to minimize eutrophication. Chemical coagulation with FeCl3 and AlSO4 is commonly used for removing phosphorous and suspended solids in wastewater treatment. The capacity of these coagulants for creating particle aggregations depends on the characteristics and chemistry of the treated wastewater, such as the ionic strength or mixing conditions. Marine water has a higher ionic strength than fresh or brackish water, which may be beneficial when using chemical coagulants to treat the effluents from farms operated at high salinities. The following study compared the application of FeCl3 and AlSO4, to treat the two effluents discharged from a marine land-based recirculating aquaculture system (RAS) producing salmon (Salmo salar). The aim of the study was to determine; 1) in what effluent (sludge flow vs. exchange water overflow) at the end-of-pipe treatment the coagulant application is more efficient for the removal of PO43−-P, total suspended solids (TSS), total phosphorous (TP) and total chemical oxygen demand (TCOD); and 2) the optimal coagulant dose to apply and its associated chemical sludge production. The results show that more than 89 % removal of TCOD, TSS and TP is achieved when treating the sludge flow, arguably because the sludge flow contained the largest fraction of the target masses (P and organic matter) discharged from the system. Up to 80 % of TSS removal was achieved by simple sedimentation, and with the highest coagulant dose tested, up to 95 % of TSS could be removed from the effluent. To remove 90 % of PO43−-P, FeCl3 and AlSO4 need to be dosed at a molar ratio of 2.6:1 Fe:PO43−-P and 5.7:1 Al: PO43−-P, respectively. Dosing above 90 % removal efficiency did not significantly affect removal of PO43-P and TSS, but substantially increased the volume of chemical sludge produced. Finally, FeCl3 is proposed as a better overall alternative for P removal at the end-of-pipe treatment in marine land-based RAS.

Investigating the effect of nitrate on juvenile turbot (Scophthalmus maximus) growth performance, health status, and endocrine function in marine recirculation aquaculture systems

Nitrate (NO3−), a potential toxic nitrogenous compound to aquatic animals, is distributed in aquatic ecosystems worldwide. The aim of this study was to investigate the effects of different NO3− levels on growth performance, health status, and endocrine function of juvenile turbot (Scophthalmus maximus) in recirculating aquaculture systems (RAS). Fish were exposed to 0 mg/L (control, CK), 50 mg/L (low nitrate, LN), 200 mg/L (medium nitrate, MN), and 400 mg/L (high nitrate, HN) NO3-N for 60 d in experimental RAS. Cumulative survival (CS) was significantly decreased with increasing NO3− levels in LN, MN, and HN. The lowest CS was 35% in the HN group. Growth parameters, including absolute growth rate, specific growth rate, and feed conversion rate, were significantly different in HN compared with that in the CK. Histological survey of gills and liver revealed dose-dependent histopathological damage induced by NO3− exposure and significant differences in glutamate pyruvate transaminase and glutamate oxalate transaminase in MN and HN compared with that in the CK. The hepatosomatic index in HN was significantly higher than that in the CK. Additionally, NO3− significantly increased bioaccumulation in plasma in LN, MN, and HN compared to that in the CK. Significant decreases in hemoglobin and increases in methemoglobin levels indicated reduced oxygen-carrying capacity in HN. Additionally, qRT-PCR and enzyme-linked immunosorbent assay (ELISA) were developed to investigate key biomarkers involved in the GH/IGF-1, HPT, and HPI axes. Compared with that in the CK, the abundance of GH, GHRb, and IGF-1 was significantly lower in HN, whereas GHRa did not differ between treatments. The plasma T3 level significantly decreased in LN, MN, and HN and T4 significantly decreased in HN. The CRH, ACTH, and plasma cortisol levels were significantly upregulated in HN compared with that in the CK. We conclude that elevated NO3− exposure leads to growth retardation, impaired health status, and endocrine disorders in turbot and the NO3− level for juvenile turbot culture should not exceed 50 mg/L NO3-N in RAS. Our findings indicate that endocrine dysfunction of the GH/IGF-1, HPT, and HPI axes might be responsible for growth inhibition induced by NO3− exposure.

Integration of Marine Macroalgae (Chaetomorpha maxima) with a Moving Bed Bioreactor for Nutrient Removal from Maricultural Wastewater.

Rather than direct nutrient removal from wastewaters, an alternative approach aimed at nutrient recovery from aquacultural wastewaters could enable sustainable management for aquaculture production. This study demonstrated the feasibility of cultivating marine macroalgae (Chaetomorpha maxima) with a moving bed bioreactor (MBBR-MA), to remove nitrogen and phosphorus in aquaculture wastewater as well as to produce macroalgae biomass. MBBR-MA significantly increased the simultaneous removal of nitrate and phosphate in comparison with only MBBR, resulting in an average total nitrogen (TN) and total phosphorus (TP) removal efficiency of 42.8 ± 5.5% and 83.7 ± 7.7%, respectively, in MBBR-MA while MBBR had no capacity for TN and TP removal. No chemical oxygen demand (COD) removal was detected in both reactors. Phosphorus could be a limiting factor for nitrogen uptake when N : P ratio increased. The recovered nitrogen and phosphorus resulted in a specific growth rate of 3.86%–10.35%/day for C. maxima with an uptake N : P ratio of 6. The presence of macroalgae changed the microbial community in both the biofilter and water by decreasing the relative abundance of Proteobacteria and Nitrospirae and increasing the abundance of Bacteroidetes. These findings indicate that the integration of the macroalgae C. maxima with MBBR could represent an effective wastewater treatment option, especially for marine recirculating aquaculture systems.

Denitrification performance evaluation and kinetics analysis with mariculture solid wastes (MSW) derived carbon source in marine recirculating aquaculture systems (RAS)

Biological denitrification using mariculture solid wastes (MSW) carbon source is a promising solution for removing nitrate (NO3−-N) and disposing MSW in marine recirculating aquaculture systems (RAS). To enhance denitrification performance, heating (HT), rhamnolipid (RL), alkali (AL), thermophilic bacteria (TB) pre-treated MSW acidogenic fermentation effluents were prepared as carbon sources. Profiles of soluble organics in four types of fermentation effluents were first evaluated. The highest volatile fatty acids (VFAs) yield (52.1%) was obtained from TB treated MSW after acidification. RL and TB treated MSW acidogenic fermentation effluents showed high NO3−-N removal efficiency (NRE) (around 97%). Acidogenic fermentation effluent from TB treated MSW presented a high biodegradability, with the minimum effluent chemical oxygen demand (COD) amount (35 mg/L). Denitrification kinetics parameters were also analyzed; high fraction (74.5%) of the most readily biodegradable organics (SS) demonstrated that TB treated MSW acidogenic fermentation effluent is a high-quality carbon source for enhancing denitrification.

Activated sludge denitrification in marine recirculating aquaculture system effluent using external and internal carbon sources

Stringent environmental legislation in Europe, especially in the Baltic Sea area, limits the discharge of nutrients to natural water bodies, limiting the aquaculture production in the region. Therefore, cost-efficient end-of-pipe treatment technologies to reduce nitrogen (N) discharge are required for the sustainable growth of marine land-based RAS. The following study examined the potential of fed batch reactors (FBR) in treating saline RAS effluents, aiming to define optimal operational conditions and evaluate the activated sludge denitrification capacity using external (acetate, propionate and ethanol) and internal carbon sources (RAS fish organic waste (FOW) and RAS fermented fish organic waste (FFOW)). The results show that between the evaluated operation cycle times (2, 4, and 6 h), the highest nitrate/nitrite removal rate was achieved at an operation cycle time of 2 h (corresponding to a hydraulic retention time of 2.5 h) when acetate was used as a carbon source. The specific denitrification rates were 98.7 ± 3.4 mg NO3−-N/(h g biomass) and 93.2 ± 13.6 mg NOx−-N/(h g biomass), with a resulting volumetric denitrification capacity of 1.20 kg NO3−-N/(m3 reactor d). The usage of external and internal carbon sources at an operation cycle time of 4 h demonstrated that acetate had the highest nitrate removal rate (57.6 ± 6.6 mg N/(h g biomass)), followed by propionate (37.5 ± 6.3 mg NO3−-N/(h g biomass)), ethanol (25.5 ± 6.0 mg NO3−-N/(h g biomass)) and internal carbon sources (7.7 ± 1.6–14.1 ± 2.2 mg NO3−-N/(h g biomass)). No TAN (Total Ammonia Nitrogen) or PO43- accumulation was observed in the effluent when using the external carbon sources, while 0.9 ± 0.5 mg TAN/L and 3.9 ± 1.5 mg PO43--P/L was found in the effluent when using the FOW, and 8.1±0.7 mg TAN/L and 7.3 ± 0.9 mg PO43--P/L when using FFOW. Average sulfide concentrations varied between 0.002 and 0.008 mg S2-/L when using the acetate, propionate and FOW, while using ethanol resulted in the accumulation of sulfide (0.26 ± 0.17 mg S2-/L). Altogether, it was demonstrated that FBR has a great potential for end-of-pipe denitrification in marine land-based RAS, with a reliable operation and a reduced reactor volume as compared to the other available technologies. Using acetate, the required reactor volume is less than half of what is needed for other evaluated carbon sources, due to the higher denitrification rate achieved. Additionally, combined use of both internal and external carbon sources would further reduce the operational carbon cost.