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).     

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.     

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.     

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.     

Evaluation of the impact of nitrate-nitrogen levels in recirculating aquaculture systems on concentrations of the off-flavor compounds geosmin and 2-methylisoborneol in water and rainbow trout (Oncorhynchus mykiss)

Aquatic animals raised in recirculating aquaculture systems (RAS) can develop preharvest “off-flavors” such as “earthy” or “musty” which are caused by the bioaccumulation of the odorous compounds geosmin or 2-methylisoborneol (MIB), respectively, in their flesh. Tainted aquatic products cause large economic losses to producers due to the inability to market them. Certain species of actinomycetes, a group of filamentous bacteria, have been attributed as the main sources of geosmin and MIB in RAS. Previous studies have demonstrated that certain nutritional factors can stimulate or inhibit bacterial biomass and geosmin production by certain actinomycetes. In the current study, the effects of two nitrate-nitrogen (NO₃^--N) levels (20–40 mg/L and 80–100 mg/L) on geosmin and MIB levels in culture water and the flesh of rainbow trout (Oncorhynchus mykiss) raised in RAS were monitored. Water and fish tissue samples were collected over an approximately nine-week period from six RAS, three replicates each of low and high NO₃^--N, and analyzed for geosmin concentrations using solid phase microextraction–gas chromatography–mass spectrometry. Results indicated no significant difference in geosmin concentrations in water or fish flesh between the low and high NO₃^--N RAS. Therefore, higher NO₃^--N levels that may occur in RAS will not adversely or beneficially impact geosmin-related off-flavor problems.     

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.     

絮团浓度对革胡子鲇零换水养殖效果的影响

为研究絮团浓度对革胡子鲇零换水养殖效果的影响，在不额外添加有机碳源（只利用饲料中的碳）的革胡子鲇（）养殖系统中，设置了平均絮团质量浓度为561.18 mg/L和780.41 mg/L两个处理组，比较了两实验组的水质、菌群结构、鱼生长及氮利用效率。结果表明，两种浓度絮团条件下，总氨氮（total ammonia nitrogen，TAN）和亚硝酸氮（NO₂^--N）能分别维持1.84 mg/L和1.79 mg/L以下。两处理组间pH、溶解氧（dissolved oxygen，DO）、TAN、NO₂^--N、氮素利用效率及主要生长指标无显著差异（-N）浓度（822.0 mg/L）明显高于低浓度絮团组（623.33 mg/L）。高通量测序分析菌群结构结果表明，两组间门水平的菌群组成种类及优势度无显著性差异（<0.05）。两处理组中的革胡子鲇存活率分别达到（91.11±1.53）%和（94.44±2.08）%，饲料系数为（1.41±0.18）和（1.27±0.26），特殊生长率为（2.13±0.04）%/d和（2.19±0.08）%/d，均无显著差异（>0.05）。两实验组饲料氮的利用率分别达到了72.17%和71.34%。综合以上结果认为，仅利用饲料中的碳既能维持革胡子鲇的零换水养殖且能取得较高的氮素利用效率，两种絮团浓度对革胡子鲇的生长无显著影响，高浓度絮团组中的硝化作用更明显。     

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).     

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.     

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.     

Effect of Dietary Protein Concentration on Nitrogenous Waste in Intensively Fed Catfish Ponds

Channel catfish were fed five diets containing 24, 28, 32, 36 or 40% protein in intensively stocked earthen ponds over a 141 d growing season. Mean standing crop at harvest was 7,559 kg/ha, and maximum daily feed allowance was 105 kg/ha. Dietary protein concentration had a negative linear effect on weight gain. Total ammonia-nitrogen (TAN) in pond water increased linearly as dietary protein concentration increased and was positively correlated with total protein fed. However, unionized ammonia-nitrogen (NH₃-N) was not influenced by dietary protein concentration. Dietary protein had a positive linear effect on nitrite-nitrogen (NO₁^-_- -N) concentration, which was positively correlated with total protein fed and TAN. There was no significant correlation between NO₂^-_--N and fish weight gain, although there was a significant positive correlation between NO₂^-_- -N/Cl-molar ratio in pond water and concentration of methemoglobin in the fish. Results from this study indicate that when the feeding rate is as high as 100 kg/ha/d, or 3,000 kg protein/ha/season, dietary protein concentrations of 36% and above can result in harmful concentrations of NO₂^-_--N when Cl concentration in the ponds is 2–3 mg/L. Although the NO₂^-_--N/Cl- ratio in the ponds increased to harmful levels with protein concentration of the diets, this might not be the major cause of the reduction in fish growth rate as dietary protein increased because the greatest difference in weight gain occurred at the lower protein concentrations and the greatest difference in NO₂^-_--N occurred at the higher dietary protein concentrations.     

Mathematical model for goldfish recirculating aquaculture system (GRAS)

A mathematical model is framed for a goldfish recirculating aquaculture system based on unsteady-state mass balance for prediction of the concentration of total ammonia nitrogen (TAN), nitrite-nitrogen (NO₂-N), nitrate-nitrogen (NO₃-N), dissolved oxygen (DO) and total suspended solids (TSS). The goldfish were stocked at 100 numbers per m³ of rearing water volume of 5 m³ tank capacity in the years 2009 and 2010 and the model was calibrated and validated. The recirculation flow rate was fixed at 29,000 L/day. The model parameters were estimated as k_TAN (mg of TAN generated per kg of feed): 20,000, M (mortality rate): 0.002 day⁻¹, α (percentage of feed conversion to suspended solids): 23.8, k_oxy (mg of oxygen required for fish respiration per kg of feed applied in unit time): 300,000, k_b (partial nitrification in the culture tank): 0.86 and the reaction rate constants, k₁ and k₂: 84.65 day⁻¹ and 42.03 day⁻¹ respectively and temperature growth coefficient (TGC): 5.00 × 10^-5. The model efficacy was adjudged by estimation of the coefficient of determination (R²), root mean square error (RMSE), Nash-Sutcliffe modelling efficiency (E_NS) and graphical plots between predicted and observed values.     

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。     

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.     

氨、亚硝酸盐和硝酸盐对凡纳滨对虾幼虾的毒性作用

研究了 NH₃-N、NO₂ ^- -N与 NO₃ ^- -N对凡纳滨对虾幼虾的毒性作用。获得了 NH₃-N_t(NH₃-N_m) 与 NO₂ ^- -N对体长2．4cm幼虾的 24h、48h、72h、96h之 LC₅₀值，两者对幼虾的安全质量分数分别为1．30 (0．101)mg/L和3．80mg/L。当 NH₃-N_t(NH₃-N_m)质量分数在1．3(0．101)~4．3(0．333)mg/L时，存活率为71．4% ~92．9%，体长增长率为36．3% ~57．1%，体重增长率为188．5% ~322．3%。当 NO₂ ^- -N质量分数在3．00~21．00mg/L时，成活率为75．0% ~91．7%，体长增长率为21．2% ~59．2%，体重增长率为72．0% ~311．9%。NO₃ ^- -N对体长7．37cm幼虾的亚急性毒性效应：NO₃ ^- -N的质量分数在 30~195mg/L时，成活率为 35% ~100%，体长增长率为8．5% ~20．5%，体重增长率为29．6% ~56．8% 。三态氮在一定质量分数范围内均对幼虾的存活率和生长率产生影响。     

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

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

Growth, stress response and free amino acid levels in Senegalese sole (Solea senegalensis Kaup 1858) chronically exposed to exogenous ammonia

Stressful husbandry conditions are likely to affect growth and amino acid metabolism in fish. In this study, chronic ammonia exposure was used to test the effects of a stressor on growth and amino acid metabolism of Senegalese sole juveniles. The fish were exposed for 52 days to 11.6 mg L⁻¹ [low‐TAN (L‐TAN)] or 23.2 mg L⁻¹ [high‐TAN (H‐TAN)] of total ammonia nitrogen (TAN), or to 0 mg L⁻¹ (Control). Growth in L‐TAN groups was slightly but significantly different from the Control groups [relative growth rate (RGR=0.35±0.13 and 0.52±0.23% day⁻¹ respectively)]. In H‐TAN groups, growth was severely affected (RGR=0.01±0.13% day⁻¹). Stress parameters (plasma cortisol and glucose) showed slight or no significant differences between treatments. Plasma free amino acid (FAA) concentrations were affected after H‐TAN treatment. Increases in glutamine and aspartate concentrations in H‐TAN fish suggest alterations in amino acid metabolism related to nitrogen excretion processes. Some of the changes in FAA concentrations also suggest mobilization to energy supply and synthesis of metabolites related to stress response. Therefore, Senegalese sole seem to adapt to the L‐TAN concentration tested, but the H‐TAN concentration reduced growth and affected amino acid metabolism.     

Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

海水观赏鱼居室养殖循环系统中三态氮的变化规律及硝化细菌对水质的影响

于 2005年 3月 5日 ~5月 3日连续监测了海水观赏鱼居室养殖循环系统的水质，研究该系统中三态氮的变化规律以及添加硝化细菌后对水质的影响。结果表明：1)试验初期氨氮的质量分数迅速上升， 在 1周内达到高峰(峰值2．56mg/L)，并在1．50mg/L的范围内维持 1周左右，此后迅速下降至0．01mg/L 左右，并一直维持在该水平直至试验结束。亚硝态氮的质量分数在氨氮的质量分数迅速回落时(约试验开始后 2周)呈现出直线上升的趋势，并在 3~3．5mg/L左右的水平上维持 2~3周时间(峰值为3．65 mg/L)，此后迅速下降至0．01mg/L以下，并一直维持在低水平直至试验结束。而硝酸盐的质量分数在整个试验期间基本保持稳定上升的趋势，至本试验末期，NO₃ ^- -N的质量分数达到 15mg/L左右。2)系统的生物滤器需要 4~5周左右时间才能基本成熟，即氨氮和亚硝酸氮均降至 ＜0．01mg/L，到达安全的质量分数，适合海水观赏鱼健康生长。3)添加硝化细菌的试验组，氨氮和 NO₃ ^- -N的质量分数变化与对照组基本相似，而 NO₂ ^- -N的质量分数变化与对照组明显不同。试验组从高质量分数水平迅速下降的时间比对照组提前了约 1周。     

Ammonia and nitrite toxicity to false clownfish <Emphasis Type="Italic">Amphiprion ocellaris</Emphasis>

False clownfish, Amphiprion ocellaris, is one of the most commercialized fish species in the world, highly produced to supply the aquarium market. The high stocking densities used to maximize fish production can increase ammonia and nitrite to toxic levels. In this study, A. ocellaris juveniles (1.20 ± 0.34 g) were exposed to six concentrations of ammonia ranged from 0.23 to 1.63 mg/L NH₃-N and eight concentrations of nitrite (26.3–202.2 mg/L NO₂ ^?-N). The LC₅₀- 24, LC₅₀-48, LC₅₀-72 and LC₅₀-96 h were estimated to be 1.06, 0.83, 0.75 and 0.75 mg/L for NH₃-N and 188.3, 151.01, 124.1 and 108.8 mg/L for NO₂ ^?-N. Analysis of gill lesions caused by sublethal concentrations of these nitrogenous compounds showed that both nitrogenous compounds induced tissue lesions such as hyperplasia of epithelium cells, hypertrophy of chloride cells and lamellar lifting to all concentrations tested. However, histopathological alterations were more conspicuous accordingly the increase of ammonia or nitrite in fish exposed to 0.57 mg/L NH₃-N or 100 mg/L NO₂ ^?-N. Based on our results, we recommend to avoid concentrations higher than 0.57 mg/L of NH₃-N and 25 mg/L of NO₂-N in water.