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All Journal Jurnal Riset Biologi dan Aplikasinya
Fauzi Akhbar Anugrah
Department of Biology, Malang State University, Jl. Semarang No. 5 Malang, East Java, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Immunology & microbiology Medicine & Pharmacology

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Stability of Microalgae-Bacteria Co-cultures During Storage in a Laboratory Freezer at −20°C: A Case Study of Chlorella vulgaris and an Indigenous Bacterial Consortium Diah Fitriana; Sitoresmi Prabaningtyas; Fauzi Akhbar Anugrah
Jurnal Riset Biologi dan Aplikasinya Vol. 8 No. 2 (2026)
Publisher : Universitas Negeri Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26740/jrba.v8n2.p137-145

Abstract

The development of microalgae-based biofuels through co-culture systems holds great potential; however, the scale-up process is often hindered by issues related to inoculum stability during storage. This study aimed to evaluate the effect of storage duration at -20 °C (for 2, 4, 6, 8, 10, and 12 weeks) on cell size and cell density of Chlorella vulgaris in a monoculture system compared with a co-culture system with an indigenous bacterial consortium (Bacillus paramycoides, Lysinibacillus fusiformis, and Delftia sp.). Optical density was measured using a UV-Vis spectrophotometer at 675 nm, while cell morphology was evaluated through microscopic observation in five fields of view. Statistical analysis using ANOVA revealed a significant decline in cell density over storage time (p < 0.05) throughout the 12 weeks in both systems. This structural preservation is presumed to be mediated by Extracellular Polymeric Substances (EPS) forming microalgae-bacteria flocs (MaB-Flocs) that serve as a protective physical buffer. However, viability assays conducted at week 4 showed no cell proliferation after thawing in either system, accompanied by distinct color degradation pathways. Therefore, storage at -20 °C without cryoprotectants is ineffective as a cryopreservation method and unsuitable for delaying cell density measurements, although co-cultivation effectively stabilizes cell morphology evaluations.
Co-Authors Diah Fitriana Sitoresmi Prabaningtyas