End-of-pipe denitrification using RAS effluent waste streams: Effect of C/N-ratio and hydraulic retention time

Environmentally sustainable aquaculture development requires increased nitrogen removal from recirculating aquaculture systems (RAS). In this study, removed solids from a large commercial outdoor recirculated trout farm (1000 MT year⁻¹) were explored as an endogenous carbon source for denitrification. This was done by (1) a controlled laboratory experiment on anaerobic hydrolysis of the organic matter (from sludge cones, drumfilter, and biofilter back-wash) and (2) an on-site denitrification factorial experiment varying the soluble COD (COD_S)/NO₃-N ratio from 4 to 12 at hydraulic retention times (HRT) from 50 to 170 min in simple 5.5 m³ denitrification reactors installed at the trout farm.The lab-experiments showed that the major part of the readily biodegradable organic matter was hydrolyzed within 14 days, and the hydrolysis rate was fastest the first 24 h. Organic matter from the sludge cones generated 0.21 ± 0.01 g volatile fatty acids (VFA) g⁻¹ total volatile solids (TVS), and the VFAs constituted 75% of COD_S. Analogously, 1 g TVS from the drum filter generated 0.15 ± 0.01 g VFA, constituting 68% of the COD_S. Comparison of the laboratory hydrolysis experiments and results from the on-farm study revealed as a rough estimate that potentially 17–24% of the generated VFA was lost due to the current sludge management.Inlet water to the denitrification reactors ranged in NO₃-N concentration from 8.3 to 11.7 g m⁻³ and COD_S from 52.9 to 113.4 g m⁻³ (10.0 ± 1.2 °C). The highest NO₃-N removal rate obtained was at the intermediate treatments; 91.5–124.8 g N m⁻³_reactor d⁻¹. The effect of the C/N ratio depended on the HRT. At low HRT, the variation in C/N ratio had no significant effect on NO₃-N removal rate, contrary to the effect at the high HRT. The stoichiometric ratio of COD_S/NO₃-N was 6.0 ± 2.4, ranging from 4.4 (at the high HRT) to 9.3 (at the low HRT). A simple model of the denitrification reactor developed in AQUASIM showed congruence between modeled and measured data with minor exceptions. Furthermore, this study pointed to the versatility of the NO₃-N removal pathways expressed by the bacterial population in response to changes in the environmental conditions; from autotrophic anammox activity presumably present at low C/N to dissimilatory nitrate reduction to ammonia (DNRA) at high C/N, besides the predominate “normal” heterotrophic dissimilatory nitrate reduction (denitrification).     

Discharge management in fresh and brackish water RAS: Combined phosphorus removal by organic flocculants and nitrogen removal in woodchip reactors

The current study combined P and N removal using organic flocculant chemicals and woodchip bioreactors in both freshwater and brackish water (7 ppm) recirculating aquaculture systems (RAS). The use of carbon (C) containing flocculant chemicals in the process was hypothesized to further stimulate C-demanding N removal (denitrification) in bioreactors. The trial of combined P and N removal consisted of four treatments: freshwater and brackish water RAS with and without the addition of supernatant from flocculation process to the woodchip reactor. Duplicate woodchip reactors were used per treatment and the trial was run for six weeks. 56% and 49% of P was removed from fresh and brackish sludge water, respectively. The nitrate-N (NO₃-N) removal rate was improved in the treatment when supernatant from flocculation process was used together with RAS discharge water when compared against the control. In brackish water RAS, the improvement was more pronounced (from 6.6–16.5 g NO₃-N m⁻³ d^-1) than in freshwater RAS (from 5.1–6.5 NO₃-N m⁻³ d^-1). In the freshwater bioreactors using supernatant, N was largely discharged as a nitrite-N (NO₂-N). High NO₂-N concentrations in freshwater reactors allude to incomplete denitrification reactions taking place. The results suggest that the organic flocculants did provide an additional C source for denitrification, which improved the N-removal process. However, in freshwater RAS this might have been partly due to untargeted processes such as DNRA (dissimilatory nitrate reduction to ammonium), and/or insufficient denitrification reactions taking place (excessive NO₂-N production).     

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 NO_x⁻-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 PO₄^3- 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 PO₄^3--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 PO₄^3--P/L when using FFOW. Average sulfide concentrations varied between 0.002 and 0.008 mg S^2-/L when using the acetate, propionate and FOW, while using ethanol resulted in the accumulation of sulfide (0.26 ± 0.17 mg S^2-/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.     

Denitrifying granules in a marine Upflow Anoxic Sludge Bed (UASB) reactor

Marine land-based Recirculating Aquaculture Systems (RAS) are generally perceived as environmentally friendly aquatic production systems. To promote their sustainability even further and reduce the discharge of nutrients, there is a need for cost-effective end-of-pipe treatment technologies for removing nutrients. This includes nitrate-nitrogen (NO₃⁻-N) for which well-proven technologies for freshwater systems exists, while similar technologies for saltwater systems are less advanced. Granular technology has been developed since the 1970s in wastewater treatment under the upflow anaerobic sludge bed (UASB) concept. This concept is based on the enrichment of different bacterial aggregations into a compact granule, optimizing synergistic and syntrophic bacterial processes by reducing the diffusion distance of substrates between the different bacterial consortia forming the granule. The following study examined the: 1) granular formation; and 2) nitrate removal capacity of a marine Upflow Anoxic Sludge Bed (UASB) reactor operating at different up-flow velocities (0.40–2.11 m/h). The results showed that marine denitrifying granules developed within 27 days using preconditioned rainbow trout (Oncorhynchus mykiss) organic matter waste, and that the highest specific denitrification rate (321.9 ± 13.1 mg NO₃⁻-N/g Total Volatile Suspended Solids (TVSS)/d) was found at an upflow velocity of 0.97 m/h. The marine UASB denitrifying granule reactor had a total capacity of removing 14.9 kg NO₃⁻-N/m³ reactor volume per day at a hydraulic retention time of 1.9 h, making it a strong candidate for end-of-pipe denitrification of marine RAS effluent as well as for in-line treatment in marine systems.     

End-of-pipe single-sludge denitrification in pilot-scale recirculating aquaculture systems

A step toward environmental sustainability of recirculat aquaculture systems (RAS) is implementation of single-sludge denitrification, a process eliminating nitrate from the aqueous environment while reducing the organic matter discharge simultaneously. Two 1700 L pilot-scale RAS systems each with a 85 L denitrification (DN) reactor treating discharged water and hydrolyzed solid waste were setup to test the kinetics of nitrate and COD removal. Nitrate removal and COD reduction efficiency was measured at two different DN-reactor sludge ages (high θ_X: 33–42 days and low θ_X: 17–23 days). Nitrate and total N (NO₃⁻ + NO₂⁻ + NH₄⁺) removal of the treated effluent water ranged from 73–99% and 60–95% during the periods, respectively, corresponding to an overall maximum RAS nitrate removal of approximately 75%. The specific nitrate removal rate increased from 17 to 23 mg NO₃⁻-N (g TVS d)⁻¹ and the maximal potential DN rate (measured at laboratory ideal conditions) increased correspondingly from 64–68 mg NO₃⁻-N (g TVS d)⁻¹ to 247–294 mg NO₃⁻-N (g TVS d)⁻¹ at high and low θ_X, respectively. Quantification of denitrifiers in the DN-reactors by qPCR showed only minor differences upon the altered sludge removal practice. The hydrolysis unit improved the biodegradability of the solid waste by increasing volatile fatty acid COD content 74–76%. COD reductions in the DN-reactors were 64–70%. In conclusion, this study showed that single-sludge denitrification was a feasible way to reduce nitrate discharge from RAS, and higher DN rates were induced at lower sludge age/increased sludge removal regime. Improved control and optimization of reactor DN-activity may be achieved by further modifying reactor design and management scheme as indicated by the variation in and between the two DN-reactors.     

Formaldehyde degradation in denitrifying woodchip bioreactors: Effects of temperature,concentration and hydraulic retention time

Formalin is applied in certain aquaculture systems to control parasites infestations as well as bacterial and fungal diseases. This study investigated the capacity of end-of-pipe denitrifying woodchip bioreactors to remove potentially harmful amounts of residual formaldehyde (FA) from aquaculture effluents. Formaldehyde was readily removed by experimental- and field-scale denitrifying woodchip bioreactors and the removal of FA was found to be a combination of an initial adsorption of FA to woodchip surfaces (52 ± 2.8 g FA/m³ woodchips) and microbial degradation. Volumetric FA removal rates reaching 261 ± 27 g FA/m³/d were found at FA inlet concentrations of 90 mg FA/L and hydraulic retention times (HRT) of 5 h. High FA removal efficiencies ranged from 88.3 ± 4.6–99.8 ± 0.2% found for FA inlet concentrations –up to 105 mg FA/L and HRTs between 3.4 and 15 h. Microbial FA degradation rates in woodchip bioreactors were positively correlated to temperature with a Q₁₀ value of 2.27 and a corresponding Arrhenius temperature coefficient of 1.086 for the investigated temperature range of 7–23 °C. At a commercial, outdoor recirculating aquaculture system (RAS) three full-scale woodchip compartments, achieved an average volumetric FA removal rate of 29.4 ± 0.2 g FA/m³/d and a removal efficiency of 82.5 ± 0.8% during the first 24 h following addition of FA. The results demonstrated that woodchip bioreactors are efficient in removing residual FA from RAS effluents and that nitrate removal was transiently enhanced during FA removal.     

Effect of support medium,hydraulic loading rate and plant density on water quality and growth of halophytes in marine aquaponic systems

The development of marine intensive land‐based aquaculture systems has been limited due to the absence of methods to manage saline wastewater. Aquaponic systems, although commonly applied to freshwater aquaculture, can potentially manage nutrient wastes while providing a secondary product. The aim of this study was to evaluate both the capacity for water treatment and the production requirements of two saltwater‐tolerant plant species (Sesuvium portulacastrum and Batis maritima) when grown hydroponically in a marine aquaponic system. The presence of plants was found to significantly contribute to nitrate removal, such that mean nitrate concentrations were 10.1 ± 5.4 and 12.1 ± 6.1 mg/L NO₃^?‐N in planted and unplanted treatments respectively. The use of coconut fibre as a planting medium also significantly contributed to nitrate removal, such that mean nitrate concentrations were 9.78 ± 5.4 and 12.4 ± 6.0 mg/L NO₃^?‐N in coconut fibre and expanded clay treatments respectively. Daily nitrogen removal was greatest in the coconut fibre/plants treatment, ranging from ?18% to 67%. Hydraulic loading rate, plant species and plant density did not significantly affect water quality or plant growth. The low flow/saltwort/low density treatment had the greatest mean daily nitrogen removal, ranging from 25% to 172%. The results indicate that the main nitrogen removal mechanisms were simultaneous nitrification–denitrification in the hydroponic plant beds and nitrogen removal through plant growth. This study demonstrates that marine aquaponics could be an effective way to manage nutrient removal in marine land‐based aquaculture systems.     

Effects of C/N ratio on nitrogen removal with denitrification phase after a nitrification-based biofloc aquaculture cycle

In the current study, we set up a denitrification process to remove the nitrogen pollutants, especially nitrate (NO₃^–-N), from the wastewater after a nitrification-based biofloc technology (BFT) aquaculture cycle. Five different treatments (CN0, CN1, CN2, CN4 and CN6, respectively) were used, which involved addition of extra carbohydrate with variable ratios of elementary organic carbon to NO₃^–-N by weight (C/NO₃^–-N ratio equal to 0, 1, 2, 4, and 6, respectively). With CN2, CN4, and CN6 treatments, NO₃^–-N was decreased (with increasing alkalinity) to ≤ 6.42 ± 0.30 mg·L⁻¹ and low amounts (close to zero) of nitrite (NO₂^–-N) were achieved. However, there were high concentrations of residual NO₃^–-N and/or NO₂^–-N in CN0 and CN1. CN2 achieved the best denitrification, wherein 81.00 ± 0.95% of the initial input nitrogen was removed. By fitting the equations, the highest nitrogen recycling rate (23.08 mg-N·g-C⁻¹) was achieved with a C/NO₃^–-N ratio of 4.16. Denitrifying bacteria were the dominant bacteria in all extra carbohydrate added treatment groups. Although denitrifying polyphosphate accumulating organisms contributed to the removal of phosphorus, high concentrations of residual soluble reactive phosphate (SRP) were observed in all treatment groups. Overall, extra addition of carbohydrate with C/NO₃^–-N ratio ≥ 2 is advisable for nitrogen removal, while the highest nitrogen recycling rate will be achieved with a ratio of 4.16.     

Heterotrophic denitrification of aquaculture effluent using fluidized sand biofilters

The ability to consistently and cost-effectively reduce nitrate-nitrogen loads in effluent from recirculating aquaculture systems would enhance the industry's environmental stewardship and allow improved facility proximity to large markets in sensitive watersheds. Heterotrophic denitrification technologies specifically employing organic carbon found in aquaculture system waste offer a unique synergy for treatment of land-based, closed-containment production outflows. For space-efficient fluidized sand biofilters to be used as such denitrification reactors, system parameters (e.g., influent dissolved oxygen and carbon to nitrogen ratios, C:N) must be evaluated to most effectively use an endogenous carbon source. The objectives of this work were to quantify nitrate removal under a range of C:Ns and to explore the biofilter bacterial community using three replicated fluidized sand biofilters (height 3.9 m, diameter 0.31 m; fluidized sand volume plus biofilm volume of 0.206 m³) operated at a hydraulic retention time of 15 min and a hydraulic loading rate of 188 L/min m² at The Conservation Fund Freshwater Institute in Shepherdstown, West Virginia, USA. Nitrate reduction was consistently observed during the biofilter study period (26.9 ± 0.9% removal efficiency; 402 ± 14 g NO₃-N/(m³ biofilter d)) although nitrite-N and total ammonium nitrogen concentrations slightly increased (11 and 13% increases, respectively). Nitrate removal efficiency was correlated with carbonaceous oxygen demand to nitrate ratios (R² > 0.70). Nitrate removal rates during the study period were moderately negatively correlated with influent dissolved oxygen concentration indicating it may be possible the biofilter hydraulic retention time was too short to provide optimized nitrate removal. It is reasonable to assume that the efficiency of nitrate removal across the fluidized sand biofilters could be substantially increased, as long as organic carbon was not limiting, by increasing biofilter bed depths (to 6–10 m), and thus hydraulic retention time. These findings provide a low-cost yet effective technology to remove nitrate-nitrogen from effluent waters of land-based closed-containment aquaculture systems.     

Wood chips and wheat straw as alternative biofilter media for denitrification reactors treating aquaculture and other wastewaters with high nitrate concentrations

This study evaluated wood chips and wheat straw as inexpensive and readily available alternatives to more expensive plastic media for denitrification processes in treating aquaculture wastewaters or other high nitrate waters. Nine 3.8-L laboratory scale reactors (40 cm packed height × 10 cm diameter) were used to compare the performance of wood chips, wheat straw, and Kaldnes plastic media in the removal of nitrate from synthetic aquaculture wastewater. These upflow bioreactors were loaded at a constant flow rate and three influent NO₃–N concentrations of 50, 120, and 200 mg/L each for at least 4 weeks, in sequence. These experiments showed that both wood chips and wheat straw produced comparable denitrification rates to the Kaldnes plastic media. As much as 99% of nitrate was removed from the wastewater of 200 mg NO₃–N/L influent concentration. Pseudo-steady state denitrification rates for 200 mg NO₃–N/L influent concentrations averaged (1360 ± 40) g N/(m³ d) for wood chips, (1360 ± 80) g N/(m³ d) for wheat straw, and (1330 ± 70) g N/(m³ d) for Kaldnes media. These values were not the maximum potential of the reactors as nitrate profiles up through the reactors indicated that nitrate reductions in the lower half of the reactors were more than double the averages for the whole reactor. COD consumption per unit of NO₃–N removed was highest with the Kaldnes media (3.41–3.95) compared to wood chips (3.34–3.64) and wheat straw (3.26–3.46). Effluent ammonia concentrations were near zero while nitrites were around 2.0 mg NO₂–N/L for all reactor types and loading rates. During the denitrification process, alkalinity and pH increased while the oxidation–reduction potential decreased with nitrate removal.

Wood chips and wheat straw lost 16.2% and 37.7% of their masses, respectively, during the 140-day experiment. There were signs of physical degradation that included discoloration and structural transformation. The carbon to nitrogen ratio of the media also decreased. Both wood chips and wheat straw can be used as filter media for biological denitrification, but time limitations for the life of both materials must be considered.     

Comparing denitrification rates and carbon sources in commercial scale upflow denitrification biological filters in aquaculture

Aerobic biological filtration systems employing nitrifying bacteria to remediate excess ammonia and nitrite concentrations are common components of recirculating aquaculture systems (RAS). However, significant water exchange may still be necessary to reduce nitrate concentrations to acceptable levels unless denitrification systems are included in the RAS design. This study evaluated the design of a full scale denitrification reactor in a commercial culture RAS application. Four carbon sources were evaluated including methanol, acetic acid, molasses and Cerelose™, a hydrolyzed starch, to determine their applicability under commercial culture conditions and to determine if any of these carbon sources encouraged the production of two common “off-flavor” compounds, 2-methyisoborneol (MIB) or geosmin. The denitrification design consisted of a 1.89 m³ covered conical bottom polyethylene tank containing 1.0 m³ media through which water up-flowed at a rate of 10 lpm. A commercial aquaculture system housing 6 metric tonnes of Siberian sturgeon was used to generate nitrate through nitrification in a moving bed biological filter. All four carbon sources were able to effectively reduce nitrate to near zero concentrations from influent concentrations ranging from 11 to 57 mg/l NO₃–N, and the maximum daily denitrification rate was 670–680 g nitrogen removed/m³ media/day, regardless of the carbon source. Although nitrite production was not a problem once the reactors achieved a constant effluent nitrate, ammonia production was a significant problem for units fed molasses and to a less extent Cerelose™. Maximum measured ammonia concentrations in the reactor effluents for methanol, vinegar, Cerelose™ and molasses were 1.62 ± 0.10, 2.83 ± 0.17, 4.55 ± 0.45 and 5.25 ± 1.26 mg/l NH₃–N, respectively. Turbidity production was significantly increased in reactors fed molasses and to a less extent Cerelose™. Concentrations of geosmin and MIB were not significantly increased in any of the denitrification reactors, regardless of carbon source. Because of its very low cost compared to the other sources tested, molasses may be an attractive carbon source for denitrification if issues of ammonia production, turbidity and foaming can be resolved.     

Evaluating the chronic effects of nitrate on the health and performance of post-smolt Atlantic salmon Salmo salar in freshwater recirculation aquaculture systems

Commercial production of Atlantic salmon smolts, post-smolts, and market-size fish using land-based recirculation aquaculture systems (RAS) is expanding. RAS generally provide a nutrient-rich environment in which nitrate accumulates as an end-product of nitrification. An 8-month study was conducted to compare the long-term effects of “high” (99 ± 1 mg/L NO₃-N) versus “low” nitrate-nitrogen (10.0 ± 0.3 mg/L NO₃-N) on the health and performance of post-smolt Atlantic salmon cultured in replicate freshwater RAS. Equal numbers of salmon with an initial mean weight of 102 ± 1 g were stocked into six 9.5 m³ RAS. Three RAS were maintained with high NO₃-N via continuous dosing of sodium nitrate and three RAS were maintained with low NO₃-N resulting solely from nitrification. An average daily water exchange rate equivalent to 60% of the system volume limited the accumulation of water quality parameters other than nitrate. Atlantic salmon performance metrics (e.g. weight, length, condition factor, thermal growth coefficient, and feed conversion ratio) were not affected by 100 mg/L NO₃-N and cumulative survival was >99% for both treatments. No important differences were noted between treatments for whole blood gas, plasma chemistry, tissue histopathology, or fin quality parameters suggesting that fish health was unaffected by nitrate concentration. Abnormal swimming behaviors indicative of stress or reduced welfare were not observed. This research suggests that nitrate-nitrogen concentrations ≤ 100 mg/L do not affect post-smolt Atlantic salmon health or performance under the described conditions.     

褐煤的碳缓释特征及其对池塘底泥脱氮作用的影响

厌氧氨氧化和反硝化作用是底泥生物脱氮的主要过程,碳源是调控厌氧氨氧化和反硝化作用的关键因子。本研究以褐煤为对象,对褐煤的静态碳释情况及其对池塘底泥中脱氮作用的影响进行了研究。结果显示,褐煤在室温条件下的碳释放规律符合二级动力学方程,具备作为反硝化碳源的可行性;在脱氮实验中,发现褐煤对底泥上覆水体中的亚硝酸盐氮(NNO₂^--N)的去除具有促进作用,NNO₂^--N的去除率随褐煤浓度的增加而升高,当褐煤质量浓度为40 g/L时,N\${\rm{O}}_2^ - $\-N去除率最高达99.61%,此时硝酸盐氮(NO₃^--N)的浓度也最低;同时发现,水体中氨氮(NH₄⁺-N)氧化的最适褐煤质量浓度为10 g/L,其去除率达99.39%;对底泥中的厌氧氨氧化菌群进行Illumina高通量测序发现,其中浮霉菌门占比最大(39.6%~71.8%),优势菌属为Candidatus Brocadia (13.9%~35.8%)和Desulfovibrio (17.1%~34.8%),添加褐煤组Candidatus Scalindua菌属比例高于未添加组;荧光定量PCR得出,随着褐煤质量浓度升高,底泥中的反硝化菌丰度呈增长趋势,而厌氧氨氧化菌丰度则低于无褐煤添加组,表明添加褐煤对底泥反硝化有促进作用,而对厌氧氨氧化有一定的抑制作用。研究表明,褐煤具备作为反硝化碳源的条件,可用于池塘养殖底泥脱氮作用。     

Anaerobic digestion of solid waste in RAS: effect of reactor type on the biochemical acidogenic potential (BAP) and assessment of the biochemical methane potential (BMP) by a batch assay

Anaerobic digestion is a way to utilize the potential energy contained in solid waste produced in recirculating aquaculture systems (RASs), either by providing acidogenic products for driving heterotrophic denitrification on site or by directly producing combustive methane. In this study the biochemical acidogenic potential of solid waste from juvenile rainbow trout was evaluated by measuring the yield of volatile fatty acids (VFA) during anaerobic digestion by batch or fed-batch reactor operation at hydrolysis time (HT)/hydraulic retention time (HRT) of 1, 5, or 10 days (and for batch additional 14 and 20 days) in continuously stirred tank reactors. Generally, the VFA yield increased with time and no effect of the reactor type used was found within the time frame of the experiment. At 10 days HT or 10 days HRT the VFA yield reached 222.3 ± 30.5 and 203.4 ± 11.2 mg VFA g⁻¹ TVS₀ (total volatile solids at day 0) in batch and fed-batch reactor, respectively. For the fed-batch reactor, increasing HRT from 5 to 10 days gained no significant additional VFA yield. Prolonging the batch reactor experiment to 20 days increased VFA production further (273.9 ± 1.6 mg VFA g⁻¹ TVS₀, n = 2). After 10 days HT/HRT, 16.8–23.5% of total Kjeldahl N was found as TAN and 44.3–53.0% of total P was found as ortho-phosphate. A significant difference between reactor types was detected for the phosphorous dissolution at 5 days HT/HRT as a relatively steep increase (of a factor 2–3) in ortho-P content occurred in fed-batch reactors but similar steep increase was only notable after 10 days HT for batch reactors. No differences between reactor types at the other HT/HRT were recorded for P as well as (for all HT/HRT for) N. Based on this study a HRT of approximately 5 days would be recommended for the design of an acidogenic continuously stirred reactor tank in a RAS single-sludge denitrification set-up. The biochemical methane potential of the sludge was estimated to 318 ± 29 g CH₄ g⁻¹ TVS₀ by a batch assay and represented a higher utility of the solid waste when comparing the methane yield with the VFA yield (in COD units). This points toward a technological challenge of ultimately increase the acidogenic output to match the methane yield as both products are formed from the same source.     

The “self cleaning inherent gas denitrification-reactor” for nitrate elimination in RAS for pike perch (Sander lucioperca) production

Denitrification reactors have proven their functionality in commercial recirculation aquaculture systems (RAS). Nevertheless, clogging occurs due to the low hydraulic loads necessary to accomplish anoxic conditions for a successful denitrification process in RAS, which hampers the adjustment of stable working conditions within fixed bed denitrification reactors. Reactors working on the basis of activated sludge demand careful hydraulic control and/or complex configurations for sludge retention.To develop a low-maintenance denitrification reactor, an enclosed moving bed filter, driven by recirculation of the inherent, oxygen poor gas was designed. A Self cleaning Inherent gas Denitrification reactor (SID-reactor) of 0.65 m³, which offered a moving bed volume of 0.39 m³ was connected with a RAS of semi-industrial scale for pike perch (Sander lucioperca) production. This species indicates suboptimal environmental conditions (as e.g. NO₃-N concentrations above approximately 68 mg l⁻¹) by prompt reduction of the feed intake. In different experimental series, the SID-reactor was operated with denatured ethanol, methanol, acetic acid or glycerin as carbon sources and changing operational modes.Clogging was prevented by a 40 second inherent gas recirculation twice an hour, which provided continuous, maintenance free operation with marginal energy demand. With inlet (RAS) and outlet NO₃-N concentrations in the range of 49 mg l⁻¹ and 12 mg l⁻¹, mean denitrification rates of 199 g to 235 g NO₃-N per m³ moving bed volume and day were determined for all tested carbon sources. Negative effects on the feed intake of the reared pike perch were detected with all carbon sources except methanol. Changing the mode of operation to continuous circulation of the filter bed at inlet NO₃-N concentrations of 26 mg l⁻¹, the denitrification performance reached 451 g NO₃-N per m³ moving bed volume and day. The SID-reactor allowed for the reduction of freshwater exchange in the pike perch RAS from 600 l to 70 l (−88%) and the sodium bicarbonate buffer from 182 g to 31 g (−83%) per kg of administered food. The easy and reliable operation of the SID-reactor could help to establish controlled denitrification as a routine purification step in RAS.     

生物强化移动床生物膜反应器处理水产养殖循环水初步研究

利用自制的硝化细菌菌剂促进移动床生物膜反应器(Moving bed biofilm reactor,MBBR)的挂膜启动,分析不同载体氨氮负荷、碳氮比条件下反应器运行状况,并进一步进行了实验室模拟循环水养殖草金鱼实验。结果显示,利用自制硝化菌剂能够完成整个移动床反应器的启动过程,在接种15 d后使循环出水氨氮稳定在1 mg/L以下。单位体积载体氨氮负荷实验表明,MBBR能够在100 mg TAN/(L填料·d)条件下,使出水满足一般水产养殖水质要求(氨氮0.5 mg/L,亚硝氮0.1 mg/L)。进水碳氮比在1以内时MBBR能够稳定高效运行。在实验室模拟循环水养殖过程中,经菌剂强化的MBBR能维持循环出水氨氮低于0.5 mg/L,亚硝氮低于0.05 mg/L。     

Larviculture of the painted river prawn Macrobrachium carcinus in different culture systems

The objective of this study was to evaluate different hatchery systems used for the larviculture of the Macrobrachium carcinus based on survival, larval development and production of post-larvae. The experimental culture was carried out in three phases designated as Phase I (Zoea VI to VIII – Z_{VI – VIII}), Phase II (Zoea VIII to X – Z_{VIII – X}), and Phase III (Zoea X to PL – Z_{X – PL}), with densities of 30, 27.5 and 25 larvae / L, respectively. The M. carcinus larvae (Z_VI) were reared in four culture systems, two being open (Greenwater – GW and Clearwater – CW) and two being closed (Biofloc – BFT and Bio-filter – RAS), distributed in twelve 10 L plastic containers, filled with 20 ppt brackish water, equipped with constant aeration, and water circulated by air lift and heated with thermostat (∼30 °C). The GW treatment was maintained with Chlorophyceae algae in the density of 3–5 × 10⁵ cells/mL. In the CW, the water was previously filtered through a 5 μm mesh screen, sterilized with 10 ppm active chlorine and, dechlorinated with vitamin C and subjected to aeration for 24 h. The BFT received water rich in bioflocs that was matured prior to the experiment and used molasses as a source of organic carbon. In the RAS, the culture water circulated through an external “Dry-Wet” biological filter. The feeding was carried out ad libitum four times daily, alternating a wet diet formula with a commercial diet, which was supplemented with newly hatched Artemia nauplii at a rate of 40–50 per larvae/day. Temperature, dissolved oxygen and pH were monitored daily and the salinity two times per week. Total ammonia, nitrite, nitrate, orthophosphate, alkalinity, total suspended solids, chlorophyll-a, COD and BOD were also analyzed. The best water quality (P < 0.05) was obtained in the RAS, with 0.49 (±0.38), 0.23 (±0.22), and 9.0 (±1.5) mg/L of TAN, NO₂-N and NO₃-N, respectively. In the GW, the nitrogen species showed high fluctuations and higher concentrations at 2.32 (±1.68), 3.53 (±3.53) and 18.2 (±12.9) mg / L of TAN, NO₂-N and NO₃-N, respectively. Considering the three phases (Z_{VI – PL}), the overall survival was 0.03, 1.97, 2.23 and 17.32 % for the BFT, CW, GW and RAS, respectively. When considering the phases separately, the survival in Phase I (Z_{VI – VIII}) was highest in the GW system at 58.7 % while the RAS was the highest in Phases II (Z_{VIII – X}) and III (Z_{X – PL}) at 70.6 % and 60.3 %, respectively. The BFT showed 8.4 (±3.5) PL/L, which was higher (P < 0.05) than that obtained in the RAS (2.8 ± 1.2 PL/L) and the GW (1.3 ± 1.1 PL/L) and similar to that obtained in the CW (5.6 ± 2.0 PL/L). Thus, the larviculture for the M. carcinus may be optimized by adopting a multiphase management strategy, which the intermediate larval stages (Z_{VI – IX}) are reared in the GW system and the final stages (Z_{X – PL}) are reared in the BFT system.     

Depuration system flushing rate affects geosmin removal from market-size Atlantic salmon Salmo salar

Common off-flavor compounds including geosmin (GSM) bioaccumulate in fish cultured in recirculating aquaculture systems (RAS) resulting in unpalatable fillets that are objectional to consumers. Most RAS facilities relocate fish from grow-out tanks to separate depuration systems with increased water flushing to remediate pre-harvest off-flavors, but certain aspects of this procedure have not been optimized including characterization of water exchange rates that effectively diminish off-flavor. To this end, a study was carried out to evaluate the effects of flushing rate and associated depuration system hydraulic retention time (HRT) on GSM removal from Atlantic salmon Salmo salar originally produced in a semi-commercial scale freshwater RAS. Twenty-six fish (5−7 kg each) were stocked into twelve replicate depuration systems operated with system HRTs of 2.4, 4.6, and 11.3-h, respectively (N = 4). Geosmin was assessed at intervals in system water and fish flesh over a 10-day feed withholding period. Waterborne GSM concentration was affected by flushing rate and associated system HRT (P < 0.05). Depuration systems operated with an 11.3-h HRT had greater waterborne GSM levels at 3, 6, and 10 days post-stocking compared to 2.4 and 4.6-h HRT. A similar trend was generally reflected in salmon flesh. Residual GSM levels were successively higher in fillets on Day 6 from depuration systems with increasingly longer HRT. Geosmin levels were greatest in salmon flesh from the 11.3-h HRT treatment on Day 10, but fillet GSM between the 2.4 and 4.6-h HRT was similar. This research indicates that lowest residual GSM is achieved in water and Atlantic salmon flesh in depuration systems with increased flushing and shorter HRT, i.e., 2.4–4.6-h under conditions of this study. Selection of optimal flushing rate to remediate off-flavor from RAS-produced Atlantic salmon may also be dictated by water and energy use metrics and site-specific water availability among other factors.     

A novel approach to denitrification processes in a zero-discharge recirculating system for small-scale urban aquaculture

This paper presents an innovative process to solve the nitrate build-up problem in recirculating aquaculture systems (RAS). The novel aspects of the process lie in a denitrification bioreactor system that uses solid cotton wool as the primary carbon source and a unique degassing chamber. In the latter, the water is physically stripped of dissolved gaseous O₂ (by means of a Venturi vacuum tube), and the subsequent denitrification becomes more efficient due to elimination of the problems of oxygen inhibition of denitrification and aerobic consumption of cotton wool. The cotton wool medium also serves as a physical barrier that traps organic particles, which, in turn, act as an additional carbon source for denitrification. Operation in the proposed system gives an extremely low C/N ratio of 0.82 g of cotton wool/g of nitrate N, which contributes to a significant reduction of biofilter volume. The additional advantage of using solid cotton wool as the carbon source is that it does not release organic residuals into the liquid to be recycled. Operation of the system over a long period consistently produced effluents with low nitrate levels (below 10 mg N/l), and there was only a very small need to replace system water. The overall treatment scheme, also incorporating an aerobic nitrification biofilter and a granular filtration device, produced water of excellent quality, i.e., with near-zero levels of nitrite and ammonia, a sufficiently high pH for aquaculture, and low turbidity. The proposed system thus provides a solution for sustainable small-scale, urban aquaculture operation with a very high recovery of water (over 99%) and minimal waste disposal.     

Denitrifying bioreactor inflow manifold design for treatment of aquacultural wastewater

Recirculating aquaculture systems (RAS) facilities subject to point-source effluent regulations need to implement cost-effective N remediation for their wastewater outflows. Relatively low-cost denitrifying “woodchip” bioreactors can effectively remove N from aquaculture effluents for at least one year, but questions remain about bioreactor lifespan for aquacultural wastewaters. Four pilot-scale bioreactors (L × W × D; 3.8 × 0.76 × 0.76 m), two with a conventional single distribution inflow manifold and two with an experimental multiple-header, feed-forward distribution manifold, were operated over 784 d to observe second-year N removal performance and to determine if the manifold design can influence bioreactor effectiveness. The study also quantified performance metrics for chemical oxygen demand, total suspended solids, and phosphorus. Manifold style did not have notable impact on bioreactor performance when treating wastewater under the facilities’ normal operating conditions, but the multiple distribution style demonstrated an 11 % increase in nitrate and 12 % increase in total suspended solids removal efficiency over the single distribution manifold toward the end of the study when bioreactors treated higher strength wastewater. Additionally, bioreactor performance in both manifold designs decreased from an average of 92 % total suspended solids removal efficiency under normal operating conditions to <76 % when treating the high-strength wastewater. The bioreactors provided N removal rates of 17−25 g NO₃-N m⁻³ d⁻¹ during the second year of study, demonstrating woodchip bioreactors can effectively treat aquaculture effluent for at least two years without major detrimental impacts due to clogging.