This study examined the performance of three independently operated denitrifying moving bed biofilm reactors (MBBRs) in a zero-exchange marine recirculating aquaculture system (RAS) stocked with European seabass (Dicentrarchus labrax). A semi-automated control strategy was applied to foster spontaneous denitrification. Process automation consisted of a pulsed carbon supply and an inflow of nitrate-rich, aerated process water controlled by the oxidation-reduction potential (ORP) in the MBBR. Carbon dosing frequency was adjusted manually if the process produced unwanted products (i.e., nitrite or ammonia). OPR-controlled inflow stimulated bacterial activities in the MBBRs until inflow reached the pre-set maximum at a hydraulic retention time (HRT) of 0.75 h. This allowed for a quick start-up of the denitrification processes in spite of high initial variability of process water inflow and of nitrate removal efficiency (NRE). A start-up with glycerol did not induce a stable denitrification process; however, after the process had been established with acetate, glycerol promoted efficient denitrification with NRE close to one. The successive application of the two carbon sources resulted in a high nitrate removal rate (NRR) of 2 kg nitrate-N m−3 day−1 in the biofilters. This diminished the concentration of nitrate-nitrogen (nitrate-N) in the RAS (volume 9 m3) from 176 to 36 g m−3 in 42 days with biofilters comprising only 1% of the RAS volume. The implications for the development of an automated denitrification process are discussed.
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Hydraulic retention time
Moving bed biofilm reactor
Nitrate removal rate
Nitrate removal efficiency
Recirculating aquaculture system
Badiola M, Mendiola D, Bostock J (2012) Recirculating aquaculture systems (RAS) analysis: main issues on management and future challenges. Aquac Eng 51:26–35. https://doi.org/10.1016/j.aquaeng.2012.07.004
Balderston WL, Sieburth JMN (1976) Nitrate removal in closed-system aquaculture by columnar denitrification. Appl Environ Microbiol 32:808–818
Bodík I, Blstáková A, Sedlácek S, Hutnan M (2009) Biodiesel waste as source of organic carbon for municipal WWTP denitrification. Bioresour Technol 100:2452–2456. https://doi.org/10.1016/j.biortech.2008.11.050
Burgin AJ, Hamilton SK (2007) Have we overemphasized the role of denitrification in aquatic ecosystems? A review of nitrate removal pathways. Front Ecol Environ 5:89–96. https://doi.org/10.1890/1540-9295(2007)5[89:HWOTRO]2.0.CO,2
Cyplik P, Juzwa W, Marecik R, Powierska-Czarny J, Piotrowska-Cyplik A, Czarny J, Drożdżyńska A, Chrzanowski L (2013) Denitrification of industrial wastewater: influence of glycerol addition on metabolic activity and community shifts in a microbial consortium. Chemosphere 93:2823–2831. https://doi.org/10.1016/j.chemosphere.2013.09.083
Davidson J, Good C, Welsh C, Summerfelt S (2011) The effects of ozone and water exchange rates on water quality and rainbow trout Oncorhynchus mykiss performance in replicated water recirculating systems. Aquac Eng 44:80–96. https://doi.org/10.1016/j.aquaeng.2011.04.001
Davidson J, Good C, Welsh C, Summerfelt ST (2014) Comparing the effects of high vs. low nitrate on the health, performance and welfare of juvenile rainbow trout Oncorhynchus mykiss within water recirculating aquaculture systems. Aquac Eng 59:30–40. https://doi.org/10.1016/j.aquaeng.2014.01.003
FAO (2016) Contributing to food security and nutrition for all. The state of world fisheries and aquaculture 2016. Rome. 200pp.
Ge S, Peng Y, Wang S, Lu CX, Zhu Y (2012) Nitrite accumulation under constant temperature in anoxic denitrification process: the effects of carbon sources and COD/NO(3)-N. Bioresour Technol 114:137–143. https://doi.org/10.1016/j.biortech.2012.03.016
Gutierrez-Wing MT, Malone RF, Rusch KA (2012) Evaluation of polyhydroxybutyrate as a carbon source for recirculating aquaculture water denitrification. Aquacultural Engineering 51:36–43. https://doi.org/10.1016/j.aquaeng.2012.07.002
Hamlin HJ, Michaels JT, Beaulaton CM, Graham WF, Dutt W, Steinbach P, Losordo TM, Schrader KK, Main KL (2008) Comparing denitrification rates and carbon sources in commercial scale upflow denitrification biological filters in aquaculture. Aquac Eng 38:79–92. https://doi.org/10.1016/j.aquaeng.2007.11.003
Han Z, Wu W, Zhu J, Chen Y (2008) Oxidization–reduction potential and pH for optimization of nitrogen removal in a twice-fed sequencing batch reactor treating pig slurry. J Biosyst Eng 99:273–281. https://doi.org/10.1016/j.biosystemseng.2007.09.013
Hardison AK, Algar CK, Giblin AE, Rich JJ (2015) Influence of organic carbon and nitrate loading on partitioning between dissimilatory nitrate reduction to ammonium (DNRA) and N2 production. Geochim Cosmochim Acta 164:146–160. https://doi.org/10.1016/j.gca.2015.04.049
He Q, Zhang D, Main K, Feng C, Ergas SJ (2018) Biological denitrification in marine aquaculture systems: a multiple electron donor microcosm study. Bioresour Technol 263:340–349. https://doi.org/10.1016/j.biortech.2018.05.018
Kiemer MCB, Black KD, Lussot D, Bullock AM, Ezzi I (1995) The effects of chronic and acute exposure to hydrogen sulphide on Atlantic salmon (Salmo salar L.). Aquacult 135:311–327. https://doi.org/10.1016/0044-8486(95)01025-4
Kraft B, Tegetmeyer HE, Sharma R, Klotz MG, Ferdelman TG, Hettich RL, Geelhoed JS, Strous M (2014) Nitrogen cycling. The environmental controls that govern the end product of bacterial nitrate respiration. Science (New York, NY) 345:676–679. https://doi.org/10.1126/science.1254070
Lee NM, Welander T (1996) The effect of different carbon sources on respiratory denitrification in biological wastewater treatment. J Ferment Bioeng 82:277–285. https://doi.org/10.1016/0922-338X(96)88820-9
Lee PG, Lea RN, Dohmann E, Prebilsky W, Turk PE, Ying H, Whitson JL (2000) Denitrification in aquaculture systems: an example of a fuzzy logic control problem. Aquac Eng 23:37–59. https://doi.org/10.1016/S0144-8609(00)00046-7
Lewis WM, Morris DP (1986) Toxicity of nitrite to fish: a review. Trans Am Fish Soc 115:183–195. https://doi.org/10.1577/1548-8659(1986)115<183:TONTF>2.0.CO,2
Li B, Irvin S (2007) The comparison of alkalinity and ORP as indicators for nitrification and denitrification in a sequencing batch reactor (SBR). Biochem Eng J 34:248–255. https://doi.org/10.1016/j.bej.2006.12.020
Li X, Blancheton JP, Liu Y, Triplet S, Michaud L (2014) Effect of oxidation-reduction potential on performance of European sea bass (Dicentrarchus labrax) in recirculating aquaculture systems. Aquacult Int 22:1263–1282. https://doi.org/10.1007/s10499-013-9745-3
Lu H, Chandran K, Stensel D (2014) Microbial ecology of denitrification in biological wastewater treatment. Water Res 64:237–254. https://doi.org/10.1016/j.watres.2014.06.042
Luccarini L, Pulcini D, Sottara D, Di Cosmo R, Canziani R (2017) Monitoring denitrification by means of pH and ORP in continuous-flow conventional activated sludge processes. Des Water Treat 61:319–325 DWt 61:319–325. https://doi.org/10.5004/dwt.2017.11119
Lupatsch I, Kissil GW, Sklan D (2001) Optimization of feeding regimes for European sea bass Dicentrarchus labrax: a factorial approach. Aquacult 202:289–302. https://doi.org/10.1016/S0044-8486(01)00779-7
Lupatsch I, Santos GA, Schrama JW, Verreth JAJ (2010) Effect of stocking density and feeding level on energy expenditure and stress responsiveness in European sea bass Dicentrarchus labrax: a factorial approach. Aquacult 298:245–250
Martins CIM, Eding EH, Verdegem MCJ, Heinsbroek LTN, Schneider O, Blancheton JP, d'Orbcastel R, Verreth JAJ (2010) New developments in recirculating aquaculture systems in Europe: a perspective on environmental sustainability. Aquac Eng 43:83–93. https://doi.org/10.1016/j.aquaeng.2010.09.002
Mota VC, Nilsen TO, Gerwins J, Gallo M, Ytteborg E, Baeverfjord G, Kolarevic J, Summerfelt ST, Terjesen BF (2019) The effects of carbon dioxide on growth performance welfare and health of Atlantic salmon post-smolt (Salmo salar) in recirculating aquaculture systems. Aquacult 498:578–586. https://doi.org/10.1016/j.aquaculture.2018.08.075
Moussavi G, Jafari SJ, Yaghmaeian K (2015) Enhanced biological denitrification in the cyclic rotating bed reactor with catechol as carbon source. Bioresour Technol 189:266–272. https://doi.org/10.1016/j.biortech.2015.04.019
Müller-Belecke A, Zienert S, Thürmer C, Kaufhold S, Spranger U (2013) The “self cleaning inherent gas denitrification-reactor” for nitrate elimination in RAS for pike perch (Sander lucioperca) production. Aquac Eng 57:18–23. https://doi.org/10.1016/j.aquaeng.2013.06.001
Oh J, Silverstein JA (1999) Acetate limitation and nitrite accumulation during denitrification. J Environ Eng 125:234–242. https://doi.org/10.1061/(ASCE)0733-9372(1999)125:3(234)
Orellana J, Waller U, Wecker B (2014) Culture of yellowtail kingfish (Seriola lalandi) in a marine recirculating aquaculture system (RAS) with artificial seawater. Aquac Eng 58:20–28. https://doi.org/10.1016/j.aquaeng.2013.09.004
Park JY, Yoo YJ (2009) Biological nitrate removal in industrial wastewater treatment: which electron donor we can choose. Appl Microbiol Biotechnol 82:415–429. https://doi.org/10.1007/s00253-008-1799-1
Pham VP, Nakano A, van der Star WRL, Heimovaara TJ, van Paassen LA (2018) Applying MICP by denitrification in soils: a process analysis. Environ Geotech 5:79–93. https://doi.org/10.1680/jenge.15.00078
Quispe CAG, Coronado CJR, Carvalho JA Jr (2013) Glycerol: production consumption prices characterization and new trends in combustion. Renew Sust Energ Rev 27:475–493. https://doi.org/10.1016/j.rser.2013.06.017
Randall DJ, Tsui TKN (2002) Ammonia toxicity in fish. Mar Pollut Bull 45:17–23
Robiana L, Corraze G, Aguirre P, Banc D, Melcion JP, Kaushik S (1999) Digestibility, postprandial ammonia excretion and selected plasma metabolites in European sea bass (Dicentrarchus labrax) fed pelleted or extruded diets with or without wheat gluten. Aquacult 179:45–56
Sauthier N, Grasmick A, Blancheton JP (1998) Biological denitrification applied to a marine closed aquaculture system. Water Res 32:1932–1938. https://doi.org/10.1016/S0043-1354(97)00406-5
Siikavuopio SI, Sæther BS (2006) Effects of chronic nitrite exposure on growth in juvenile Atlantic cod Gadus morhua. Aquacult 255:351–356. https://doi.org/10.1016/j.aquaculture.2005.11.058
Strohm TO, Griffin B, Zumft WG, Schink B (2007) Growth yields in bacterial denitrification and nitrate ammonification. Appl Environ Microbiol 73:1420–1424. https://doi.org/10.1128/AEM.02508-06
Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41:100–180
Timmons MB, Ebeling JM (2010) Recirculating Aquaculture. NRAC Publication No 401-2010. Cayuga Aqua Ventures, Ithaca NY
Torno J, Naas C, Schroeder JP, Schulz C (2018) Impact of hydraulic retention time backflushing intervals and C/N ratio on the SID-reactor denitrification performance in marine RAS. Aquacult 496:112–122. https://doi.org/10.1016/j.aquaculture.2018.07.004
Torrans EL, Clemens HP (1982) Physiological and biochemical effects of acute exposure of fish to hydrogen sulfide. Comp Biochem Physiol C: Comp phar 71:183–190
van Bussel CGJ, Schroeder JP, Wuertz S, Schulz C (2012) The chronic effect of nitrate on production performance and health status of juvenile turbot (Psetta maxima). Aquacult 326:163–167. https://doi.org/10.1016/j.aquaculture.2011.11.019
van den Berg EM, Rombouts JL, Kuenen JG, Kleerebezem R, van Loosdrecht MCM (2017) Role of nitrite in the competition between denitrification and DNRA in a chemostat enrichment culture. AMB Express 7:91. https://doi.org/10.1186/s13568-017-0398-x
Viana MB, Freitas AV, Leitão RC, Pinto GAS, Santaella ST (2012) Anaerobic digestion of crude glycerol: a review. Environ Technol Rev 1:81–92. https://doi.org/10.1080/09593330.2012.692723
Waller U (2001) Tank culture and recirculating systems. Kenneth D Black (Hg.): Environmental impacts of aquaculture. Sheffield Academy Press (Sheffield biological sciences), Sheffield, pp 99–127
Waller U, Orellana J, Sander M (2007) The control of water quality and hygienic conditions in aquaculture recirculation systems (RAS): the use of foam fractionation and ozone. Proceedings of the IOA/IUVA Joint World Congress on Ozone and Ultraviolet Technologies. Los Angeles USA pp. 1916–1936
Wolfe AJ (2005) The acetate switch. Microbiol Mol Biol Rev 69:12–50. https://doi.org/10.1128/MMBR.69.1.12-50.2005
Yang X, Wang S, Zhou L (2012) Effect of carbon source C/N ratio nitrate and dissolved oxygen concentration on nitrite and ammonium production from denitrification process by Pseudomonas stutzeri D6. Bioresour Technol 104:65–72. https://doi.org/10.1016/j.biortech.2011.10.026
Yilmaz H, Corraze G, Panserat S, Eroldogan OT (2016) Effects of alternate feeding with different lipid sources on fatty acid composition and bioconversion in European sea bass (Dicentrarchus labrax). Aquacul 464:28–23
Zhu T, Dittrich M (2016) Carbonate precipitation through microbial activities in natural environment and their potential in biotechnology: a review. Front Bioeng Biotechnol 4. https://doi.org/10.3389/fbioe.2016.00004
Zhu SM, Deng YL, Ruan YJ, Guo XS, Shi MM, Shen JZ (2015) Biological denitrification using poly(butylene succinate) as carbon source and biofilm carrier for recirculating aquaculture system effluent treatment. Bioresour Technol 192:603–610. https://doi.org/10.1016/j.biortech.2015.06.021
This study was a part of the project “Modellbasierte innovative Regelungsstrategien für biologische Prozesse der Lebensmittelindustrie (MARE)” funded by the BMBF-Support program for young engineers 2013 (AZ: 13FH004I3) and the project "Microbial Stabilization Technologies (MicStaTech)" funded by the COFASP ERA-NET partners, who received funding from the European Union’s Seventh Framework Programme for research, technological development, and demonstration under grant agreement no. 321553. O.S.-Z. was supported by an ERASMUS grant to pursue his master degree at the htw saar.
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Stavrakidis-Zachou, O., Ernst, A., Steinbach, C. et al. Development of denitrification in semi-automated moving bed biofilm reactors operated in a marine recirculating aquaculture system. Aquacult Int 27, 1485–1501 (2019). https://doi.org/10.1007/s10499-019-00402-5
- Moving bed biofilm reactor
- Nitrate removal
- ORP control
- Recirculating aquaculture system