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Microbial Community Structure and Nitrogen in a Saline Filter Bioreactor Inoculated with Shrimp Pond Sludge

Zulkarnaini Zulkarnaini, Muhammad Varrel, Zelvi Indira, Norihisa Matsuura

Abstract


Understanding the composition of microbial communities is essential for optimizing anammox-based nitrogen removal in saline wastewater environments. This study examined the microbial diversity and ecological structure within a filter bioreactor inoculated with shrimp pond sludge and operated under saline conditions (30.1–33.0 ppt) and fed with synthetic seawater containing 70 mg-N/L of ammonium and nitrite with hydraulic retention time 24 h. Over 175 days of operation, the reactor maintained stable nitrogen removal, with peak ammonium conversion and nitrogen removal efficiencies of 46.25% and 44.33%, respectively. High-throughput 16S rRNA gene sequencing illuminated a diverse microbial community, dominated by Candidatus Brocadia (8.07%), alongside significant representations of Candidatus Jettenia (0.88%). The microbial consortium also included key nitrifying bacteria such as Nitrosomonas and Nitrospira, indicating synergistic interactions in nitrogen transformation. These taxa play key functional roles in nitrogen transformation, biofilm stability, and adaptation to saline, anoxic environments. Phylogenetic analysis showed close affiliations between amplicon sequence variants and recognized anammox species, such as Candidatus Brocadia sinica and Candidatus Jettenia asiatica. The detection of freshwater-associated anammox genera in a saline system highlights their ecological adaptability and potential application in saline wastewater treatment. Overall, this study provides insights into microbial consortia that drive anammox processes in engineered saline environments and supports the development of biological nitrogen removal strategies for marine and coastal applications.

Keywords



[1]    Y. Zhou, Y. Zhu, J. Zhu, C. Li, and G. Chen, “A comprehensive review on wastewater nitrogen removal and its recovery processes,” International Journal of Environmental Research and Public Health, vol. 20, no. 4, p. 3429, 2023, doi: 10.3390/ijerph20043429.

[2]    L. Zhang, L. Jiang, J. Zhang, J. Li, and Y. Peng, “Enhancing nitrogen removal through directly integrating anammox into mainstream wastewater treatment: Advantageous, issues and future study,” Bioresource Technology, vol. 362, p. 127827, Oct. 2022, doi: 10.1016/j.biortech.2022. 127827.

[3]    S. Liu, C. Cai, F. Sun, M. Ma, T. An, and C. Chen, “Advanced nitrogen removal of landfill leachate treatment with anammox process: A critical review,” Journal of Water Process Engineering, vol. 58, p. 104756, Feb. 2024, doi: 10.1016/j.jwpe.2023.104756.

[4]    D. Parde, M. Behera, R. R. Dash, and P. Bhunia, “A review on anammox processes: Strategies for enhancing bacterial growth and performance in wastewater treatment,” International Biodeterioration & Biodegradation, vol. 191, p. 105812, Jun. 2024, doi: 10.1016/j.ibiod.2024.105812.

[5]    J. A. C. Roques et al., “Tolerance of the marine anammox candidatus scalindua to high nitrate concentrations: Implications for recirculating aquaculture systems,” Water (Switzerland), vol. 16, no. 24, Dec. 2024, doi: 10.3390/w16243705.

[6]    I. N. Ismail et al., “Anammox process for aquaculture wastewater treatment: Operational condition, mechanism, and future prospective,” Water Science and Technology, vol. 86, no. 12, pp. 3093–3112, Dec. 2022, doi: 10.2166/wst. 2022.403.

[7]    X. Ji, Y. Wang, and P.-H. Lee, “Evolution of microbial dynamics with the introduction of real seawater portions in a low-strength feeding anammox process,” Applied Microbiology and Biotechnology, vol. 104, pp. 5593–5604, Apr. 2020, doi: 10.1007/s00253-020-10598-9.

[8]    Zulkarnaini, Q. Yujie, R. Yamamoto-Ikemoto, and N. Matsuura, “One-stage nitritation/ anammox process using a biofilm reactor with two-inflow,” Journal of Water and Environment Technology, vol. 16, no. 2, pp. 106–114, 2018, doi: 10.2965/jwet.17-050.

[9]    G. Gumelar, E. N. Zainuddin, and Z. Zulkarnaini, “Fast start-up marine anammox process using intensive shrimp pond solid waste as inoculum in filter bioreactor,” Journal of Sustainability Science and Management, vol. 19, no. 1, pp. 55–62, 2024.

[10]  N. Yokota et al., “High-rate nitrogen removal from waste brine by marine anammox bacteria in a pilot-scale UASB reactor,” Applied Microbiology and Biotechnology, vol. 102, no. 3, pp. 1501–1512, Feb. 2018, doi: 10.1007/s00253-017-8663-0.

[11]  F. Lulrahman, S. Silvia, and Z. Zulkarnaini, “Nitrogen removal by anammox process using sludge from muara penjalinan of padang city as inoculum,” Jurnal Teknologi Lingkungan, vol. 23, no. 2, pp. 143–150, 2022, doi: 10.29122/ jtl.v23i2.5284.

[12]  I. N. Ismail, “Development of anaerobic ammonium oxidation (anammox) process reactor for aquaculture wastewater treatment,” Universiti Malaysia Terengganu, 2022.

[13]  G. Gumelar, E. Zainuddin, and Z. Zulkarnaini, “Anaerobic ammonium oxidation performance in shrimp pond wastewater treatment,” Andalasian International Journal of Applied Science, Engineering and Technology, vol. 02, no. 02, pp. 51–56, 2022, doi: 10.25077/aijaset. v2i1.41.

[14]  T. Lotti, R. Kleerebezem, C. Lubello, and M. C. M. C. M. M. van Loosdrecht, “Physiological and kinetic characterization of a suspended cell anammox culture,” Water Research, vol. 60, pp. 1–14, Sep. 2014, doi: 10.1016/j.watres.2014.04. 017.

[15]  L. Zhang, M. Liu, S. Zhang, Y. Yang, and Y. Peng, “Integrated fixed-biofilm activated sludge reactor as a powerful tool to enrich anammox biofilm and granular sludge,” Chemosphere, vol. 140, pp. 114–118, Dec. 2015, doi: 10.1016/ j.chemosphere.2015.02.001.

[16]  APHA, Standard Methods for the Examination of Water and Wastewater, 23rd ed., Washington, DC: American Public Health Association, 2017.

[17]  K. Koike, G. J. Smith, R. Yamamoto-Ikemoto, S. Lücker, and N. Matsuura, “Distinct comammox Nitrospira catalyze ammonia oxidation in a full-scale groundwater treatment bioreactor under copper limited conditions,” Water Research, vol. 210, Feb. 2022, doi: 10.1016/j.watres.2021. 117986.

[18]  W. Ludwig et al., “ARB: A software environment for sequence data,” Nucleic Acids Research, vol. 32, no. 4, pp. 1363–1371, 2004, doi: 10.1093/nar/gkh293.

[19]  L. V. Duc, B. Song, H. Ito, T. Hama, M. Otani, and Y. Kawagoshi, “High growth potential and nitrogen removal performance of marine anammox bacteria in shrimp-aquaculture sediment,” Chemosphere, vol. 196, pp. 69–77, Apr. 2018, doi: 10.1016/j.chemosphere.2017.12. 159.

[20]  Y. Kawagoshi, Y. Nakamura, H. Kawashima, K. Fujisaki, K. Furukawa, and A. Fujimoto, “Enrichment of marine anammox bacteria from seawater-related samples and bacterial community study,” Water Science and Technology, vol. 61, no. 1, pp. 119–126, 2010, doi: 10.2166/wst. 2010.796.

[21]  M. Ali, D. R. Shaw, M. Albertsen, and P. E. Saikaly, “Comparative genome-centric analysis of freshwater and marine anammox cultures suggests functional redundancy in nitrogen removal processes,” Frontiers in Microbiology, vol. 11, Jul. 2020, doi: 10.3389/fmicb.2020.01637.

[22]  Y. Kawagoshi, K. Fujisaki, Y. Tomoshige, K. Yamashiro, and Y. Qiao, “Temperature effect on nitrogen removal performance and bacterial community in culture of marine anammox bacteria derived from sea-based waste disposal site,” Journal of Bioscience and Bioengineering, vol. 113, no. 4, pp. 515–520, Apr. 2012, doi: 10.1016/j.jbiosc.2011.11.024.

[23]  A. Oren, G. M. Garrity, and O. Garrity, “Valid publication of the names of forty-two phyla of prokaryotes,” International Journal of Systematic and Evolutionary Microbiology, vol. 71, p. 5056, 2021, doi: 10.1099/ijsem.0.005056.

[24]  X. D. Wang, Y. Y. Wang, S. K. Song, W. G. Wang, M. Wu, and D. L. Wang, “Impact of salinity on the performance and microbial community of anaerobic ammonia oxidation (anammox) using 16s rRNA high-throughput sequencing technology,” Global Nest Journal, vol. 19, no. 3, pp. 377–388, Nov. 2017, doi: 10.30955/GNJ.002207.

[25]  V. F. Dsane, S. An, M. K. Shahid, and Y. Choi, “From freshwater anammox bacteria (FAB) to marine anammox bacteria (MAB): A stepwise salinity acclimation process,” Science of the Total Environment, vol. 796, Nov. 2021, doi: 10.1016/J.SCITOTENV.2021.148753.

[26]  V. F. Dsane, S. An, T. Oh, J. Hwang, Y. Choi, and Y. Choi, “Saline conditions effect on the performance and stress index of anaerobic ammonium oxidizing (anammox) bacteria,” Chemosphere, vol. 267, Mar. 2020, doi: 10.1016/J.CHEMOSPHERE.2020.129227.

[27]  A. D. Pereira, A. Cabezas, C. Etchebehere, C. A. de L. Chernicharo, and J. C. de Araújo, “Microbial communities in anammox reactors: A review,” Environmental Technology Reviews, vol. 6, no. 1 pp. 74–73, Jan. 2017, doi: 10.1080/21622515.2017.1304457.

[28]  D. J. Steele, D. J. Franklin, and G. J. C. Underwood, “Protection of cells from salinity stress by extracellular polymeric substances in diatom biofilms,” Biofouling, vol. 30, no. 8, pp. 987–998, Sep. 2014, doi: 10.1080/08927014.  2014.960859.

[29]  A. Banerjee, S. Sarkar, S. Cuadros-Orellana, and R. Bandopadhyay, “Exopolysaccharides and biofilms in mitigating salinity stress: The biotechnological potential of halophilic and soil-inhabiting PGPR microorganisms,” Springer, Cham, 2019, pp. 133–153, doi: 10.1007/978-3-030-18975-4_6.

[30]  W. A. Alhoqail, “Exopolysaccharide-Producing PGPR: Mechanisms for alleviating salinity-induced plant stress,” Polish Journal of Environmental Studies, pp. 1–18, Mar. 2025, doi: 10.15244/PJOES/196747.

[31]  Y. Zhou, Q. Wen, C. Pang, Z. Wang, and Z. Chen, “Performance and adaptation mechanisms of Anammox granular sludge under salinity stress: Role of EPS, microbial community and functional genes,” Chemical Engineering Journal, vol. 514, p. 163185, Jun. 2025, doi: 10.1016/J.CEJ.2025.163185.

[32]  L. V. Duc, B. Song, H. Ito, T. Hama, M. Otani, and Y. Kawagoshi, “High growth potential and nitrogen removal performance of marine anammox bacteria in shrimp-aquaculture sediment,” Chemosphere, vol. 196, pp. 69–77, Apr. 2018, doi: 10.1016/j.chemosphere.2017.12. 159.

[33]  N. Yokota et al., “High-rate nitrogen removal from waste brine by marine anammox bacteria in a pilot-scale UASB reactor,” Applied Microbiology and Biotechnology vol. 102, no. 3, pp. 1501–1512, 2018, doi: 10.1007/s00253-017-8663-0.

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DOI: 10.14416/j.asep.2026.02.004

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