Skip to main content

The influence of wastewater pretreatment, attachment material, and inoculation strategy on the growth of target algal species in cultivated biofilms

Abstract

Microalgal biofilms are promising sources of sustainable biofuels and bioproducts because of their minimal growth requirements, ability to sequester CO2, and high biomass concentrations. However, strategies to enhance the growth of product-producing species within the biofilm communities must be investigated to optimize the performance and profitability of algal biofilm cultivation systems. Quantitative polymerase chain reaction assays were used to investigate how wastewater, biofilm attachment material, and inoculation strategies influence the abundance of Chlorella vulgaris and Scenedesmus obliquus in biofilms. A 5.6-fold increase in biofilm dry weight was achieved by pretreating reactors with wastewater and using cotton biofilm attachment materials compared to that on non-pretreated polycarbonate attachment materials. Furthermore, depending on the inoculation strategy and the presence of wastewater, the composition of the algal community in biofilms differed from their corresponding suspensions and from the inoculums, highlighting the importance of analyzing the species in algal biofilms in cultivation systems. This study represents the first application of quantitative polymerase chain reaction to estimate cell abundance of C. vulgaris and S. obliquus in cultivated algal biofilms and provides insights into the optimization of algal biofilm cultivation systems for the commercialization of algal biofuels or other bioproducts.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Data availability

All data generated or analyzed during this study are included in this published article [and its supplementary information files].

References

  • Blanken W, Janssen M, Cuaresma M et al (2014) Biofilm growth of Chlorella sorokiniana in a rotating biological contactor based photobioreactor. Biotechnol Bioeng 111:2436–2445

    CAS  Article  Google Scholar 

  • Borowitzka MA (2013) High-value products from microalgae-their development and commercialisation. J Appl Phycol 25:743–756

    CAS  Article  Google Scholar 

  • Cavanaugh SE, Bathrick AS (2018) Direct PCR amplification of forensic touch and other challenging DNA samples: a review. Forensic Sci Int Genet 32:40–49

    CAS  Article  Google Scholar 

  • Chen X, Kong L, Wang X, Tian S, Xiong Y (2015) Accelerated start-up of moving bed biofilm reactor by using a novel suspended carrier with porous surface. Bioprocess Biosyst Eng 38:273–285

    CAS  Article  Google Scholar 

  • Cheng P, Wang Y, Liu T, Liu D (2017) Biofilm attached cultivation of Chlorella pyrenoidosa is a developed system for swine wastewater treatment and lipid production. Front Plant Sci 8:1594

    Article  Google Scholar 

  • Cheung SL, Allen DG, Short SM (2020) Specific quantification of Scenedesmus obliquus and Chlorella vulgaris in mixed-species algal biofilms. Bioresour Technol 295:122251

    CAS  Article  Google Scholar 

  • Christenson LB, Sims RC (2012) Rotating algal biofilm reactor and spool harvester for wastewater treatment with biofuels by-products. Biotechnol Bioeng 109:1674-1684 

    CAS  Article  PubMed  Google Scholar 

  • Cole JJ (1982) Interactions between bacteria and algae in aquatic ecosystems. Annu Rev Ecol Syst 13:291–314

    Article  Google Scholar 

  • Deantes-Espinosa VM, Zhang T-Y, Wang X-X, Wu Y, Dao G-H, Hu H-Y (2019) Attached cultivation of Scenedesmus sp: LX1 on selected solids and the effect of surface properties on attachment. Front Environ Sci Eng 13:57

    Article  Google Scholar 

  • Ferris MJ, Muyzer G, Ward DM, Spring O (1996) Denaturing gradient gel electrophoresis profiles of 16S rRNA-defined populations inhabiting a hot spring microbial mat community. Appl Environ Microbiol 62:340–346

    CAS  Article  Google Scholar 

  • Gao F, Yang Z-H, Li C, Zeng G-M, Ma D-H, Zhou L (2015) A novel algal biofilm membrane photobioreactor for attached microalgae growth and nutrients removal from secondary effluent. Bioresour Technol 179:8–12

    CAS  Article  Google Scholar 

  • Genin SN, Aitchison SJ, Allen DG (2014) Design of algal film photobioreactors: Material surface energy effects on algal film productivity, colonization and lipid content. Bioresour Technol 155:136–143

  • Gross M, Henry W, Michael C, Wen Z (2013) Development of a rotating algal biofilm growth system for attached microalgae growth with in situ biomass harvest. Bioresour Technol 150:195–201

    CAS  Article  Google Scholar 

  • Gross M, Zhao X, Mascarenhas V, Wen Z (2016) Effects of the surface physico-chemical properties and the surface textures on the initial colonization and the attached growth in algal biofilm. Biotechnol Biofuels 9:38

    Article  Google Scholar 

  • Hodges A, Fica Z, Wanlass J, VanDarlin J, Sims R (2017) Nutrient and suspended solids removal from petrochemical wastewater via microalgal biofilm cultivation. Chemosphere 174:46–48

    CAS  Article  Google Scholar 

  • Huang Y, Zheng Y, Li J, Liao Q, Fu Q, Xia A, Fu J, Sun Y (2018) Enhancing microalgae biofilm formation and growth by fabricating microgrooves onto the substrate surface. Bioresour Technol 261:36–43

    CAS  Article  Google Scholar 

  • Irving TE, Allen DG (2011) Species and material considerations in the formation and development of microalgal biofilms. Appl Microbiol Biotechnol 92:283–294

    CAS  Article  Google Scholar 

  • Ji C, Wang J, Liu T, Tag TÁ (2015) Aeration strategy for biofilm cultivation of the microalga Scenedesmus dimorphus. Biotechnol Lett 37:1953–1958

    CAS  Article  Google Scholar 

  • Katevatis C, Fan A, Klapperich CM (2017) Low concentration DNA extraction and recovery using a silica solid phase. PLoS One 12:e0176848

    Article  Google Scholar 

  • Kiperstok AC, Sebestyén P, Podola B, Melkonian M (2017) Biofilm cultivation of Haematococcus pluvialis enables a highly productive one-phase process for astaxanthin production using high light intensities. Algal Res 21:213–222

    Article  Google Scholar 

  • Lanlan Z, Lin C, Junfeng W, Yu C, Xin G, Zhaohui Z, Tianzhong L (2015) Attached cultivation for improving the biomass productivity of Spirulina platensis. Bioresour Technol 181:136–142

  • Lee SH, Oh HM, Jo BH, Lee SA, Shin SY, Kim HS, Lee SH, Ahn CY (2014) Higher biomass productivity of microalgae in an attached growth system, using wastewater. J Microbiol Biotechnol 24:1566–1573

    CAS  Article  Google Scholar 

  • Li J, Helmerhorst EJ, Leone CW, Troxler RF, Yaskell T, Haffajee AD, Socransky SS, Oppenheim FG (2004) Identification of early microbial colonizers in human dental biofilm. J Appl Microbiol 97:1311–1318

    CAS  Article  Google Scholar 

  • Liu T, Wang J, Hu Q, Cheng P, Ji B, Liu J, Chen Y, Zhang W, Chen X, Chen L, Gao L, Ji C, Wang H (2013) Attached cultivation technology of microalgae for efficient biomass feedstock production. Bioresour Technol 127:216–222

    CAS  Article  Google Scholar 

  • Molina Grima E, Belarbi EH, Acién Fernández FG, Robles Medina A, Chisti Y (2003) Recovery of microalgal biomass and metabolites: process options and economics. Biotechnol Adv 20:491–515

    CAS  Article  Google Scholar 

  • Morden CW, Delwiehe CF, Kuhsel M, Palmer JD (1992) Gene phylogenies and the endosymbiotic origin of plastids. BioSystems 28:75–90

    CAS  Article  Google Scholar 

  • Mujtaba G, Lee K (2017) Treatment of real wastewater using co-culture of immobilized Chlorella vulgaris and suspended activated sludge. Water Res 120:174–184

    CAS  Article  Google Scholar 

  • Ozkan A, Berberoglu H (2013a) Adhesion of algal cells to surfaces. Biofouling 29:469–482

  • Ozkan A, Berberoglu H (2013b) Cell to substratum and cell to cell interactions of microalgae. Colloids Surfaces B 112:302–309

  • Peng J, Kumar K, Gross M, Kunetz T, Wen Z (2020a) Removal of total dissolved solids from wastewater using a revolving algal biofilm reactor. Water Environ Res 92:766–778

    CAS  Article  Google Scholar 

  • Peng YY, Gao F, Yang HL, Wu HW, Li C, Lu MM, Yang ZY (2020b) Simultaneous removal of nutrient and sulfonamides from marine aquaculture wastewater by concentrated and attached cultivation of Chlorella vulgaris in an algal biofilm membrane photobioreactor (BF-MPBR). Sci Total Environ 725:138524

    CAS  Article  Google Scholar 

  • Piltz B, Melkonian M (2018) Immobilized microalgae for nutrient recovery from source-separated human urine. J Appl Phycol 30:421–429

    CAS  Article  Google Scholar 

  • Rajendran A, Hu B (2016) Mycoalgae biofilm: development of a novel platform technology using algae and fungal cultures. Biotechnol Biofuels 9:1–13

    CAS  Article  Google Scholar 

  • Schenk PM, Thomas-Hall SR, Stephens E, Marx UC, Mussgnug JH, Posten C, Kruse O, Hankamer B (2008) Second generation biofuels: high-efficiency microalgae for biodiesel production. BioEnergy Res 1:20–43

    Article  Google Scholar 

  • Scott NG, J., Stewart Aitchison J, Grant Allen D (2014) Design of algal film photobioreactors: Material surface energy effects on algal film productivity colonization and lipid content. Bioresour Technol 155:136-143

    CAS  Article  PubMed  Google Scholar 

  • Sekar R, Venugopalan VP, Satpathy KK, Nair KVK, Rao VNR (2004) Laboratory studies on adhesion of microalgae to hard substrates. Hydrobiologia 512:109–116

    Article  Google Scholar 

  • Shen Y, Xu X, Zhao Y, Lin X (2014) Influence of algae species, substrata and culture conditions on attached microalgal culture. Bioprocess Biosyst Eng 37:441–450

    CAS  Article  Google Scholar 

  • Spolaore P, Joannis-Cassan C, Duran E, Isambert A (2006) Commercial applications of microalgae. J Biosci Bioeng 101:87–96

    CAS  Article  Google Scholar 

  • Stein J (ed) (1973) Handbook of phycological methods: culture methods and growth measurements. Cambridge University Press, Cambridge

    Google Scholar 

  • Toth CRA, Luo F, Bawa N, Webb J, Guo S, Dworatzek S, Edwards EA (2021) Anaerobic benzene biodegradation linked to growth of highly specific bacterial clades. Environ Sci Technol 55:7970–7980

    CAS  Article  Google Scholar 

  • Wang JH, Zhuang LL, Xu XQ, Deantes-Espinosa VM, Wang XX, Hu HY (2018) Microalgal attachment and attached systems for biomass production and wastewater treatment. Renew Sustain Energy Rev 92:331–342

    Article  Google Scholar 

  • Xu XQ, Wang JH, Zhang TY, Dao GH, Wu GX, Hu HY (2017) Attached microalgae cultivation and nutrients removal in a novel capillary-driven photo-biofilm reactor. Algal Res 27:198–205

    Article  Google Scholar 

  • Yang J, Gou Y, Fang F, Guo J, Liu L, Zhou Y, Ma H (2018) Potential of wastewater treatment using a concentrated and suspended algal-bacterial consortium in a photo membrane bioreactor. Chem Eng J 335:154–160

  • Ye Y, Huang Y, Xia A, Fu Q, Liao Q, Zeng W, Zheng Y, Zhu X (2018) Optimizing culture conditions for heterotrophic-assisted photoautotrophic biofilm growth of Chlorella vulgaris to simultaneously improve microalgae biomass and lipid productivity. Bioresour Technol 270:80–87

    CAS  Article  Google Scholar 

  • Zhang Q, Wang L, Yu Z, Zhou T, Gu Z, Huang Q, Xiao B, Zhou W, Ruan R, Liu Y (2020) Pine sawdust as algal biofilm biocarrier for wastewater treatment and algae-based byproducts production. J Clean Prod 256:120449

    CAS  Article  Google Scholar 

Download references

Acknowledgements

Thank you to Line Lomheim and Dr. Shen Guo for qPCR support.

Funding

Financial support for this project was received from the Natural Sciences and Engineering Research Council (NSERC) of Canada Strategic Grants program [grant number 463037–14] and Discovery Grants program [grant number 2016–05524]. Scholarship support was received from the Ontario Graduate Scholarships program, the NSERC CGS program, the Queen Elizabeth II Graduate Scholarship in Science and Technology program, and the Department of Chemical Engineering and Applied Chemistry at the University of Toronto.

Author information

Authors and Affiliations

Authors

Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Samantha Cheung. The first draft of the manuscript was written by Samantha Cheung and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to D. Grant Allen.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Cheung, S.L., Short, S.M. & Allen, D.G. The influence of wastewater pretreatment, attachment material, and inoculation strategy on the growth of target algal species in cultivated biofilms. J Appl Phycol 34, 113–125 (2022). https://doi.org/10.1007/s10811-021-02637-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10811-021-02637-3

Keywords

  • Microalgae
  • Biofilm
  • Wastewater
  • Quantitative PCR
  • Scenedesmus obliquus
  • Chlorella vulgaris