Denitrifying Microorganisms on Suspended Carrier Biofilm in a Recirculating Aquaculture System of Pacific White Shrimp Litopenaeus vannamei
LI Yayuan1,2,3, ZHANG Wang1,2,3, LIU Xingguo2,3, XIAO Shuwen1,2,3, JIANG Haixin4, LU Shimin2,3
1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;
2. Fishery Machinery and Instrument Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200092, China;
3. Key Laboratory of Aquaculture Facilities Engineering, Ministry of Agriculture and Rural Affairs, Shanghai 200092, China;
4. College of Environmental Science and Engineering, Tongji University, Shanghai 200092, China
Abstract:To understand the nitrate reduction potential and nitrogen metabolism pathway character of moving bed bioreactor in a Pacific white shrimp Litopenaeus vannamei in a recirculating aquaculture system(RAS), the microbial community structure and denitrification efficiency of biofilms were analyzed on the surface of suspended carriers. Illumina sequencing results revealed that Proteobacteria(20.31%—25.36%) and Bacteroidetes(14.42%—24.28%) were dominant in all samples. At the genus level, aerobic denitrifiers such as Rhodococcus (0.28%—0.97%), Paracoccus (0.50%—0.99%) and Pseudomonas (0.15%—0.30%) were primarily responsible for nitrate reduction. Furthermore, qRT-PCR detection showed that the denitrification functional gene nirK was more abundant than that of nirS and nosZ, so the predominant denitrification bacteria were of the nirK-type.The prediction results from functional annotation of prokaryotic taxa (FAPROTAX) confirmed the presence of active heterotrophic denitrification in biofilms, the denitrifying bacteria were mainly competitive with each other and with the dominant biofilm bacteria,possibly due to the limited carbon sources. The anaerobic nitrogen removal tests showed that there was nitrate removal efficiency of up to 98.07%, without adding any carbon source. Based on the characteristics of nitrogen metabolism pathway, the partial denitrification-anammox process for shrimp aquaculture wastewater nitrogen removal was further proposed. The finding provides novel insights into comprehending of nitrogen metabolic pathways in RAS and future management, utilization of endogenous carbon sources in the biofilms, and potential denitrification functions.
李雅媛, 张旺, 刘兴国, 肖述文, 江海鑫, 陆诗敏. 凡纳滨对虾循环水养殖系统生物膜反硝化菌研究[J]. 水产科学, 2024, 43(5): 775-783.
LI Yayuan, ZHANG Wang, LIU Xingguo, XIAO Shuwen, JIANG Haixin, LU Shimin. Denitrifying Microorganisms on Suspended Carrier Biofilm in a Recirculating Aquaculture System of Pacific White Shrimp Litopenaeus vannamei. Fisheries Science, 2024, 43(5): 775-783.
[1] VAN RIJN J. Waste treatment in recirculating aquaculture systems[J]. Aquacultural Engineering,2013,53:49-56.
[2] 穆显鑫,苗德霞,孙丹妮,等.氨氮质量浓度及附着基筛选对硝化细菌氨氮净化影响[J].水产科学,2022,41(1):122-129.
[3] MENASVETA P, PANRITDAM T, SIHANONTH P, et al. Design and function of a closed, recirculating seawater system with denitrification for the culture of black tiger shrimp broodstock[J]. Aquacultural Engineering,2001,25(1):35-49.
[4] PUNGRASMI W, PLAYCHOOM C, POWTONGSOOK S. Optimization and evaluation of a bottom substrate denitrification tank for nitrate removal from a recirculating aquaculture system[J]. Journal of Environmental Sciences,2013,25(8):1557-1564.
[5] ALVES NETO I, BRANDÃO H, FURTADO P S, et al. Acute toxicity of nitrate in Litopenaeus vannamei juveniles at low salinity levels[J]. Ciência Rural,2019,49(1):1-9.
[6] HIBIYA K, TERADA A, TSUNEDA S, et al. Simultaneous nitrification and denitrification by controlling vertical and horizontal microenvironment in a membrane-aerated biofilm reactor[J]. Journal of Biotechnology,2003,100(1):23-32.
[7] BHATTACHARYA R, MAZUMDER D. Simultaneous nitrification and denitrification in moving bed bioreactor and other biological systems[J]. Bioprocess and Biosystems Engineering,2021,44(4):635-652.
[8] ZHOU X L, ZHANG Y A, LI Z H, et al. A novel two-stage anoxic/oxic-moving bed biofilm reactor process for biological nitrogen removal in a full-scale municipal WWTP:performance and bacterial community analysis[J]. Journal of Water Process Engineering,2022,50:103224.
[9] ZHU L, YUAN H Z, SHI Z, et al. Metagenomic insights into the effects of various biocarriers on moving bed biofilm reactors for municipal wastewater treatment[J]. Science of the Total Environment,2022,813:151904.
[10] YUAN Q A, WANG H Y, CHU Z S, et al. Influence of C/N ratio on MBBR denitrification for advanced nitrogen removal of wastewater treatment plant effluent[J]. Desalination and Water Treatment,2017,66:158-165.
[11] FENG L J, LUO Y Q, YANG J Y, et al. Nitrogen removal characteristics in a biofilm system for recirculating aquaculture wastewater treatment under high-salinity conditions and oligotrophic stress[J]. Journal of Environmental Engineering,2020,146(7):3-8.
[12] LU J, ZHANG Y X, WU J, et al. Nitrogen removal in recirculating aquaculture water with high dissolved oxygen conditions using the simultaneous partial nitrification, anammox and denitrification system[J]. Bioresource Technology,2020,305:123037.
[13] ZHANG Y X, LU J, WU J, et al. Potential risks of microplastics combined with superbugs:enrichment of antibiotic resistant bacteria on the surface of microplastics in mariculture system[J]. Ecotoxicology and Environmental Safety,2020,187:109852.
[14] GAO Y D, GUO L, SHAO M Y, et al. Heterotrophic denitrification strategy for marine recirculating aquaculture wastewater treatment using mariculture solid wastes fermentation liquid as carbon source:optimization of COD/NO-3-N ratio and hydraulic retention time[J]. Bioresource Technology,2020,304:122982.
[15] WANG S Y, LIU C L, WANG X X, et al. Dissimilatory nitrate reduction to ammonium (DNRA) in traditional municipal wastewater treatment plants in China:widespread but low contribution[J]. Water Research,2020,179:115877.
[16] PICAZO A, VILLAESCUSA J A, ROCHERA C, et al. Functional metabolic diversity of bacterioplankton in maritime Antarctic Lakes[J]. Microorganisms,2021,9(10):2077.
[17] 蔺凌云,尹文林,潘晓艺,等.自然微生物挂膜处理水产养殖废水的效果及微生物群落分析[J].水生生物学报,2017,41(6):1327-1335.
[18] HUANG Z T, WAN R, SONG X F, et al. Metagenomic analysis shows diverse, distinct bacterial communities in biofilters among different marine recirculating aquaculture systems[J]. Aquaculture International,2016,24(5):1393-1408.
[19] RUAN Y J, GUO X S, YE Z Y, et al. Bacterial community analysis of different sections of a biofilter in a full-scale marine recirculating aquaculture system[J]. North American Journal of Aquaculture,2015,77(3):318-326.
[20] ITAKURA M, SAEKI K, OMORI H, et al. Genomic comparison of Bradyrhizobium japonicum strains with different symbiotic nitrogen-fixing capabilities and other Bradyrhizobiaceae members[J]. The ISME Journal,2009,3(3):326-339.
[21] 孙永欣,田斌,任同军,等.微生物在海水循环水养殖系统中的作用及其应用进展[J].水产科学,2023,42(2):321-330.
[22] ZHAO W H, PENG Y Z, WANG M X, et al. Nutrient removal and microbial community structure variation in the two-sludge system treating low carbon/nitrogen domestic wastewater[J]. Bioresource Technology,2019,294:122161.
[23] CHEN S N, LI S L, HUANG T L, et al. Nitrate reduction by Paracoccus thiophilus strain LSL 251 under aerobic condition:performance and intracellular central carbon flux pathways[J]. Bioresource Technology,2020,308:123301.
[24] CHEN P Z, LI J, LI Q X, et al. Simultaneous heterotrophic nitrification and aerobic denitrification by bacterium Rhodococcus sp. CPZ24[J]. Bioresource Technology,2012,116:266-270.
[25] SONG C G, ZHAO C K, WANG Q Z, et al. Impact of carbon/nitrogen ratio on the performance and microbial community of sequencing batch biofilm reactor treating synthetic mariculture wastewater[J]. Journal of Environmental Management,2021,298:113528.
[26] SHU D T, HE Y L, YUE H, et al. Metagenomic and quantitative insights into microbial communities and functional genes of nitrogen and iron cycling in twelve wastewater treatment systems[J]. Chemical Engineering Journal,2016,290:21-30.
[27] CHEN Z, CHANG Z Q, QIAO L, et al. Effect of hydraulic retention time on solid-phase denitrification reactor in recirculating aquaculture system[J]. Aquaculture,2021, 543:736928.
[28] DENG M, DAI Z L, SENBATI Y, et al. Aerobic denitrification microbial community and function in zero-discharge recirculating aquaculture system using a single biofloc-based suspended growth reactor:influence of the carbon-to-nitrogen ratio[J]. Frontiers in Microbiology,2020,11:1760.
[29] JONES C M, STRES B, ROSENQUIST M, et al. Phylogenetic analysis of nitrite, nitric oxide, and nitrous oxide respiratory enzymes reveal a complex evolutionary history for denitrification[J]. Molecular Biology and Evolution,2008,25(9):1955-1966.
[30] YUAN H G, HUANG S B, YUAN J Q, et al. Characteristics of microbial denitrification under different aeration intensities:performance, mechanism, and co-occurrence network[J]. Science of the Total Environment,2021,754:141965.
[31] HENZE M. Wastewater treatment:biological and chemical processes[M]. 3rd ed. Berlin:Springer,2002.
[32] GAN Y L, ZHAO Q L, YE Z F. Denitrification performance and microbial diversity of immobilized bacterial consortium treating nitrate micro-polluted water[J]. Bioresource Technology,2019,281:351-358.