Effect of Temperature and Preservation Period on the Viability of Lyophilized Bacillus subtilis

Kim  Vanessa Phoane; Enty  Tjoa; Shikha Joon; Listya  Utami Karmawan; Lucky  Hartati Moehario

doi:10.35652/IGJPS.2022.12017

Authors

Kim Vanessa Phoane Faculty of Medicine and Health Sciences, Atma Jaya Catholic University of Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-6254-7189
Enty Tjoa Department of Microbiology, School of Medicine and Health Sciences, Atma Jaya Catholic University of Indonesia, Jakarta, Indonesia https://orcid.org/0000-0001-7187-4280
Shikha Joon Institutes for Systems Genetics, Frontiers Science Center for Disease-Related Molecular Network, West China Hospital, Sichuan University, Chengdu, Sichuan, China. ; iGlobal Research and Publishing Foundation, New Delhi, India. https://orcid.org/0000-0002-9178-962X
Listya Utami Karmawan Department of Biotechnology, Faculty of Biotechnology, Atma Jaya Catholic University of Indonesia, Jakarta, Indonesia https://orcid.org/0000-0002-6591-5362
Lucky Hartati Moehario Department of Microbiology, School of Medicine and Sciences, Atma Jaya Catholic University of Indonesia, Jakarta, Indonesia. https://orcid.org/0000-0003-4234-9932

DOI:

https://doi.org/10.35652/IGJPS.2022.12017

Keywords:

Bacillus subtilis, Temperature, Preservation period, Viability, Lyophilized

Abstract

Objective: We sought to study the effect of temperature and preservation period on the viability of lyophilized Bacillus subtilis. Methods: Forty-five test tubes that contained suspension of B. subtilis ATCC 6633 were incubated for 4 hours at 37°C and washed twice before freeze-drying. Next, the lyophilized bacteria were exposed to different temperatures (-20°C, 2-8°C, and 25°C) and storage periods (30 days, 120 days, and 240 days). Further, the lyophilized bacteria were evaluated for their viability using the Miles and Misra method. Finally, the colony count of B. subtilis in each group was compared and analyzed using the two-way ANOVA and the Tukey-test (post-hoc test). Results: The lyophilized B. subtilis showed the viability of 93.3% (42/45) of test tubes. All of the lyophilized B. subtilis stored at -20°C for 30, 120, and 240 days remained viable while the bacteria stored at 2-8°C and 25°C showed the viability of 93.3% (14/15) and 86.6%, respectively. The lyophilized B. subtilis stored at three group temperatures and kept for 120 and 240 days showed a significant decline in their number over the control group (p <0.05). Conclusion: In conclusion, the temperature and preservation period significantly affect the viability of the lyophilized B. subtilis. © 2022 iGlobal Research and Publishing Foundation. All rights reserved.