Box-Behnken Design-Based Optimization of Treatment Parameters for Soluble Reactive Phosphorus Removal of Synthetic Wastewater using Immobilized Spirulina platensis Beads

Authors

  • Sean Andre D. Calajate Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines
  • Francis Edric M. Robles Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines
  • Maria Francesca I. Rojas Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines
  • Tristan Josef A. Tolentino Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines
  • Angela Nicole S. Masongsong Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University – Laguna, Laguna Boulevard, LTI Spine Road, Barangays Biñan and Malamig, Biñan City, Laguna 4024, Philippines
  • John Ray C. Estrellado Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University – Laguna, Laguna Boulevard, LTI Spine Road, Barangays Biñan and Malamig, Biñan City, Laguna 4024, Philippines https://orcid.org/0000-0002-5789-1380 (unauthenticated)

DOI:

https://doi.org/10.11594/ijmaber.06.06.32

Keywords:

alginate, immobilization, soluble reactive phosphorus, Spirulina platensis, wastewater treatment

Abstract

Soluble reactive phosphorus (SRP), a bioavailable phosphorus form, contributes to over-eutrophication by stimulating uncontrolled algal growth. This study aims to determine the optimum treatment parameters for the SRP removal from synthetic wastewater using the alginate-immobilized cyanobacteria Spirulina platensis. S. platensis was immobilized in alginate beads with varying alginate concentrations (2.5%, 3%, and 3.5% w/v), and subjected to varying operation time (1, 2, and 3 days), and bead dosage (1.5, 2, and 2.5 beads/mL) for SRP removal using Box-Behnken experimental design. Resulting model indicated a strong predictive relationship with R2 = 0.9253 and p = 0.0212. Main effects of bead dosage (p = 0.01372), its quadratic effect (p = 0.01643), and its interaction with alginate concentration (p = 0.00465) were found to be statistically significant. Predicted optimum parameters (2.5% w/v alginate, 3 days, and 1.5 beads/mL) were validated and resulted in a lower SRP removal of 92.80 ± 0.73% with a percent error of 5.22% relative to a predicted SRP removal of 97.91%. Extrapolation of the prediction model to 100% outside the experimental region was verified resulting in SRP removal of 97.39 ± 0.08% with a percent error of 2.61% was achieved by adjusting the operation time to 3.4 days. The study shows promising potential of immobilized S. platensis beads in addressing over-eutrophication through significant phosphorus reduction.

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Author Biographies

  • Sean Andre D. Calajate, Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines

    Sean Andre D. Calajate is a Grade 12 student of STEM from DLSU Manila. He has experience in journalism and has presented as a delegate of the Integrated School of DLSU Manila during the 7th International Conference on Philippine and Asian Studies in 2024.

  • Francis Edric M. Robles, Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines

    Francis Edric M. Robles is a Grade 12 STEM student from DLSU Manila. He was the former Editor in Chief of his previous school. He also has a background in managing funds and project leading due to his internship at his father’s company.

  • Maria Francesca I. Rojas, Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines

    Maria Francesca I. Rojas is a Grade 12 Student from the STEM Department of DLSU Manila. She is the current President of the DLSU-SHS STEM Society, the Student Council Grade 12 STEM Representative, as well as former mentee of the 2023 STEM-Konek Program by Unilab PH.

  • Tristan Josef A. Tolentino, Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University, 2401 Taft Ave., Malate, Manila, Philippines

    Tristan Josef A. Tolentino is a Grade 12 student of the STEM Department of DLSU Manila, recognized as the 15th most outstanding student of Makati City during the Ten Outstanding Students of Makati competition last 2023. He represented DLSU Manila during the 7th International Conference on Philippine and Asian Studies.

  • Angela Nicole S. Masongsong, Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University – Laguna, Laguna Boulevard, LTI Spine Road, Barangays Biñan and Malamig, Biñan City, Laguna 4024, Philippines

    Angela Masongsong is a licensed Geologist working as a STEM Research Mentor at De La Salle University Integrated School. She obtained her B.S. Geology degree from the University of the Philippines Diliman as a DOST-SEI Merit Scholar. Her research interests are mainly scientific education, environmental monitoring, and geospatial applications. 

  • John Ray C. Estrellado, Department of Science, Technology, Engineering, and Mathematics, The Academy, De La Salle University – Laguna, Laguna Boulevard, LTI Spine Road, Barangays Biñan and Malamig, Biñan City, Laguna 4024, Philippines

    John Ray C. Estrellado is a licensed chemical engineer currently taking his Master of Science in Chemical Engineering in De La Salle University. His research interests include encapsulation technologies and hydrocolloids. He is teaching practical research courses under the Department of Science, Technology, Engineering, and Mathematics at DLSU Integrated School.

References

Abdel Hameed, M. S. (2007). Effect of algal density in bead, bead size, and bead con-centrations on wastewater nutrient re-moval. African Journal of Biotechnology, 6(10), 1185-1191. https://www.ajol.info/index.php/ajb/article/view/57139

Banerjee, S., Tiwade, P. B., Sambhav, K., Banerjee, C., & Bhaumik, S. K. (2019). Ef-fect of alginate concentration in wastewater nutrient removal using algi-nate-immobilized microalgae beads: Up-take kinetics and adsorption studies. Bio-chemical Engineering Journal, 149, 107241. https://doi.org/10.1016/j.bej.2019.107241

Bouabidi, Z. B., El-Naas, M. H., & Zhang, Z. (2018). Immobilization of microbial cells for the biotreatment of wastewater: A re-view. Environmental Chemistry Letters, 17(1), 241–257. https://doi.org/10.1007/s10311-018-0795-7

Brandão, B. C. S., Oliveira, C. Y. B., Santos, E. P., Abreu, J. L. D., Oliveira, D. W. S., Cabral da Silva, S. M. B., & Gálvez, A. O. (2023). Mi-croalgae-based domestic wastewater treatment: A review of biological aspects, bioremediation potential, and biomass production with biotechnological high-value. Environmental Monitoring and As-sessment, 195(1384). https://doi.org/10.1007/s10661-023-12031-w

Calvo-López, A., Ymbern, O., Puyol, M., & Alonso-Chamarro, J. (2021). Soluble reac-tive phosphorus determination in wastewater treatment plants by automat-ic microanalyzers. Talanta, 221, 121508. https://doi.org/10.1016/j.talanta.2020.121508

Chai, W. S., Tan, W. G., Munawaroh, H. S. H., Gupta, V. K., Ho, S. H., & Show, P. L. (2021). Multifaceted roles of microalgae in the application of wastewater bio-treatment: A review. Environmental Pol-lution, 269, 116236. https://doi.org/10.1016/j.envpol.2020.116236

Chaieb, K., Kouidhi, B., Ayed, L., Hosawi, S. B., Abdulhakim, J. A., Hajri, A., & Altayb, H. N. (2023). Enhanced textile dye removal from wastewater using natural bio-sorbent and Shewanella algae B29: Ap-plication of Box Behnken design and ge-nomic approach. Bioresource Technology, 374, 128755. https://doi.org/10.1016/j.biortech.2023.128755

Chen, X., Lee, Y., Yuan, T., Lei, Z., Adachi, Y., Zhang, Z., Lin, Y., & Van Loosdrecht, M. C. (2022). A review on recovery of extracel-lular biopolymers from flocculent and granular activated sludges: Cognition, key influencing factors, applications, and challenges. Bioresource Technology, 363, 127854. https://doi.org/10.1016/j.biortech.2022.127854

Cruz, I., Bashan, Y., Hernàndez-Carmona, G., & De-Bashan, L. E. (2013). Biological dete-rioration of alginate beads containing immobilized microalgae and bacteria during tertiary wastewater treatment. Applied Microbiology and Biotechnology, 97(22), 9847–9858. https://doi.org/10.1007/s00253-013-4703-6

de-Bashan, L. E., & Bashan, Y. (2010). Immobi-lized microalgae for removing pollutants: Review of practical aspects. Bioresource Technology, 101(6), 1611-1627. https://doi.org/10.1016/j.biortech.2009.09.043

Department of Environment and Natural Re-sources. (2021). Water quality guidelines and general effluent standards of 2016 (DENR Administrative Order No. 2021-XX, Sec. 5.3). https://www.denr.gov.ph/

Domini, M., Abbà, A., & Bertanza, G. (2022). Analysis of the variation of costs for sew-age sludge transport, recovery, and dis-posal in Northern Italy: A recent survey (2015–2021). Water Science & Technolo-gy, 85(4), 1167–1175. https://doi.org/10.2166/wst.2022.040

El-Sheekh, M., Morsi, H., & Hassan, L. (2020). Growth Enhancement of Spirulina platen-sis through Optimization of Media and Ni-trogen Sources. Egyptian Journal of Botany, 0(0), 0. https://doi.org/10.21608/ejbo.2020.27927.1487

Eroglu, E., Smith, S. M., & Raston, C. L. (2015). Application of various immobilization techniques for algal bioprocesses. In Bi-omass and Biofuels from Microalgae (pp. 19–44). Springer. https://doi.org/10.1007/978-3-319-16640-7_2

Ghaeni, M., & Roomiani, L. (2016). Effects of Spirulina, microalgae. Journal of Ad-vanced Agricultural Technologies, 3(2), 114-117. https://doi.org/10.18178/joaat.3.2.114-117

Gichana, Z., Liti, D., Drexler, S., Zollitsch, W., Meulenbroek, P., Wakibia, J., Ogello, E., Akoll, P., & Waidbacher, H. (2019). Ef-fects of aerated and non-aerated biofil-ters on effluent water treatment from a small-scale recirculating aquaculture sys-tem for Nile tilapia (Oreochromis nilot-icus L.). Die Bodenkultur Journal of Land Management Food and Environment, 70(4), 209–219. https://doi.org/10.2478/boku-2019-0019

Halim, A. A., & Haron, W. N. a. W. (2021). Im-mobilized Microalgae using Alginate for Wastewater Treatment. Pertanika Jour-nal of Science & Technology, 29(3). https://doi.org/10.47836/pjst.29.3.34

Hossain, S. M. Z., Alnoaimi, A., Razzak, S. A., Ezuber, H., Al‐Bastaki, N., Safdar, M., Al-kaabi, S., & Hossain, M. M. (2018). Multi-objective optimization of microalgae (Chlorella sp.) growth in a photobioreac-tor using Box‐Behnken design approach. The Canadian Journal of Chemical Engi-neering, 96(9), 1903–1910. https://doi.org/10.1002/cjce.23168

Hossain, S. M. Z., Sultana, N., Jassim, M. S., Coskuner, G., Hazin, L. M., Razzak, S. A., & Hossain, M. M. (2022). Soft-computing modeling and multiresponse optimization for nutrient removal process from munic-ipal wastewater using microalgae. Jour-nal of Water Process Engineering, 45, 102490. https://doi.org/10.1016/j.jwpe.2021.102490

Huno, S. K., Rene, E. R., van Hullebusch, E. D., & Annachhatre, A. P. (2018). Nitrate re-moval from groundwater: a review of natural and engineered processes. Jour-nal of Water Supply: Research and Tech-nology—AQUA, 67(8), 885-902

Karydis, M. (2013). Eutrophication assessment of coastal waters based on indicators: a literature review. Global NEST Journal, 11(4), 373–390. https://doi.org/10.30955/gnj.000626

Khatoon, H., Penz, K. P., Banerjee, S., Rahman, M. R., Minhaz, T. M., Islam, Z., Mukta, F. A., Nayma, Z., Sultana, R., & Amira, K. I. (2021). Immobilized Tetraselmis sp. for reducing nitrogenous and phosphorous compounds from aquaculture wastewater. Bioresource Technology, 338, 125529. https://doi.org/10.1016/j.biortech.2021.125529

Klokk, T. I., & Melvik, J. E. (2002). Controlling the size of alginate gel beads by use of a high electrostatic potential. Journal of Mi-croencapsulation, 19(4), 415–424. https://doi.org/10.1080/02652040210144234

Lee, B., Ravindra, P., & Chan, E. (2013). Size and shape of calcium alginate beads pro-duced by extrusion dripping. Chemical Engineering & Technology, 36(10), 1627–1642. https://doi.org/10.1002/ceat.201300230

Li, Y., Wu, X., Liu, Y., & Taidi, B. (2024). Immo-bilized microalgae: Principles, processes, and its applications in wastewater treat-ment. World Journal of Microbiology and Biotechnology, 40(150). https://doi.org/10.1007/s11274-024-03930-2

Lin, Y., & Tanaka, S. (2006). Oxygen transfer and mixing in bioreactors: A review.

Biochemical Engineering Journal, 30(1), 1–7. https://doi.org/10.1016/j.bej.2005.11.010

Maali, A., Gheshlaghi, R., & Mahdavi, M. A. (2024). Maximizing key biochemical products of Spirulina platensis: optimal light quantities and best harvesting time. OCL, 31, 21. https://doi.org/10.1051/ocl/2024019

Malone, T., & Newton, A. (2020). Effects of nu-trient pollution in marine ecosystems are compounded by human activity. Frontiers in Marine Science. https://phys.org/news/2020-08-effects-nutrient-pollution-marine-ecosystems.html

Molinuevo-Salces, B., Riaño, B., Hernández, D., & García-González, M. C. (2019). Microal-gae and wastewater treatment: Ad-vantages and disadvantages. In M. Alam & Z. Wang (Eds.), Microalgae biotechnology for development of biofuel and wastewater treatment (pp. 505–533). Springer. https://doi.org/10.1007/978-981-13-2264-8_20

Mollamohammada, S. (2020). Nitrate and herbicides removal from groundwater using immobilized algae (Doctoral disser-tation). University of Nebraska-Lincoln. https://digitalcommons.unl.edu/civilengdiss/154

Mazur, L. P., Cechinel, M. A., De Souza, S. M. U., Boaventura, R. A., & Vilar, V. J. (2018). Brown marine macroalgae as natural cat-ion exchangers for toxic metal removal from industrial wastewaters: A review. Journal of Environmental Management, 223, 215–253. https://doi.org/10.1016/j.jenvman.2018.05.086

Oldenborg, K. A., & Steinman, A. D. (2019). Im-pact of sediment dredging on sediment phosphorus flux in a restored riparian wetland. Science of the Total Environ-ment, 650, 1969-1979.

Osman, G. A., Ali, M. S., Kamel, M. M., & Amber, S. G. (2011). The role of Cladophora sp. and Spirulina platensis in the removal of microbial flora in Nile water. New York Science Journal, 4(3), 8–17, 4(3). http://www.sciencepub.net/newyork

Paerl, H. W. (2009). Controlling eutrophication along the freshwater–marine continuum: Dual nutrient (N and P) reductions are essential. Estuaries and Coasts, 32(4), 593–601. https://doi.org/10.1007/s12237-009-9158-8

Parsons, T. R., Maita, Y., & Lalli, C. M. (1984). Determination of phosphate. In Elsevier eBooks (pp. 22–25). https://doi.org/10.1016/b978-0-08-030287-4.50015-3

Porkka, T. (2021). Optimization of microalgal immobilization for cultivation in aquacul-ture wastewater (Master's thesis). Uni-versity of Eastern Finland. https://erepo.uef.fi/items/19c210d8-376e-4ee2-82f2-6071d70371bb

Purev, O., Park, C., Kim, H., Myung, E., Choi, N., & Cho, K. (2023). Spirulina platensis im-mobilized alginate beads for removal of Pb(II) from aqueous solutions. Interna-tional Journal of Environmental Research and Public Health, 20(2), 1106. https://doi.org/10.3390/ijerph20021106

Patnaik, S., Sarkar, R., & Mitra, A. (2001). Algi-nate immobilization of Spirulina platensis for wastewater treatment. Indian journal of experimental biology, 39(8), 824–826. https://pubmed.ncbi.nlm.nih.gov/12018590/

Rajasekaran, C., Ajeesh, C. P. M., Balaji, S., Shalini, M., Siva, R., Das, R., Fulzele, D. P., & Kalaivani, T. (2015). Effect of Modified Zarrouk’s Medium on Growth of Different Spirulina Strains. Walailak Journal of Sci-ence and Technology (WJST), 13(1), 67–75. https://www.researchgate.net/publication/291699334_Effect_of_Modified_Zarrouk's_Medium_on_Growth_of_Different_Spirulina_Strains

Sajid, M., Asif, M., Baig, N., Kabeer, M., Ihsanul-lah, I., & Mohammad, A. W. (2022). Car-bon nanotubes-based adsorbents: Prop-erties, functionalization, interaction mechanisms, and applications in water purification. Journal of Water Process En-gineering, 47, 102815. https://doi.org/10.1016/j.jwpe.2022.102815

Shpigel, M., Neori, A. (2007). Microalgae, Macroalgae, and Bivalves as Biofilters in Land-Based Mariculture in Israel. In: Bert, T.M. (eds) Ecological and Genetic Impli-cations of Aquaculture Activities. Meth-ods and Technologies in Fish Biology and Fisheries, vol 6. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-6148-6_24

Santos, A. F., Mendes, L. S., Alvarenga, P., Gan-do-Ferreira, L. M., & Quina, M. J. (2024). Nutrient Recovery via Struvite Precipita-tion from Wastewater Treatment Plants: Influence of Operating Parameters, Coex-isting Ions, and Seeding. Water, 16(12), 1675. https://doi.org/10.3390/w16121675

Tam, N., & Wong, Y. (2000). Effect of immobi-lized microalgal bead concentrations on wastewater nutrient removal. Environ-mental Pollution, 107(1), 145–151. https://doi.org/10.1016/s0269-7491(99)00118-9

Taqiyyah, A. M., Risjani, Y., Prihanto, A. A., Yanuhar, U., & Fadjar, M. (2022). Effect of Aquaculture Wastewater And Zarrouk in Increasing Biomass, Protein, and Carote-noids levels of Spirulina platensis. Jurnal Ilmiah Perikanan Dan Kelautan. https://doi.org/10.20473/jipk.vi.40822

Velusamy, K., Periyasamy, S., Kumar, P. S., Vo, D. V. N., Sindhu, J., Sneka, D., & Sub-hashini, B. (2021). Advanced techniques to remove phosphates and nitrates from waters: A review. Environmental Chemis-try Letters, 19, 3165–3180. https://link.springer.com/article/10.1007/s10311-021-01239-2

Vonshak, A. (1997). Spirulina platensis arthro-spira. In CRC Press eBooks. https://doi.org/10.1201/9781482272970

Wang, L., Liu, X., Li, Z., Wan, C., & Zhang, Y. (2023). Filamentous aerobic granular sludge: A critical review on its cause, im-pact, control and reuse. Journal of Envi-ronmental Chemical Engineering, 11(3), 110039. https://doi.org/10.1016/j.jece.2023.110039

Xu, S., Li, Z., Yu, S., Chen, Z., Xu, J., Qiu, S., & Ge, S. (2024). Microalgal–bacteria biofilm in wastewater treatment: Advantages, prin-ciples, and establishment. Water, 16(18), 2561. https://doi.org/10.3390/su162411196

Yang, Z., Pei, H., Han, F., Wang, Y., Hou, Q., & Chen, Y. (2018). Effects of air bubble size on algal growth rate and lipid accumula-tion using fine-pore diffuser photobiore-actors. Algal Research, 32, 293–299. https://doi.org/10.1016/j.algal.2018.04.016

You, F., Fan, Y., Tang, L., Liu, X., Jin, C., Zhao, Y., Wang, Y., & Guo, L. (2025). Optimiza-tion of Phaeodactylum tricornutum culti-vation for enhancing mariculture wastewater treatment and high value product recovery using Box–Behnken de-sign. Process Safety and Environmental Protection, 107022. https://doi.org/10.1016/j.psep.2025.107022

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Published

2025-06-24

How to Cite

Calajate, S. A. D., Robles, F. E. M., Rojas, M. F. I., Tolentino, T. J. A., Masongsong, A. N. S., & Estrellado, J. R. C. (2025). Box-Behnken Design-Based Optimization of Treatment Parameters for Soluble Reactive Phosphorus Removal of Synthetic Wastewater using Immobilized Spirulina platensis Beads. International Journal of Multidisciplinary: Applied Business and Education Research, 6(6), 3070-3092. https://doi.org/10.11594/ijmaber.06.06.32