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Comparison of various approaches to detect algal culture contamination: a case study of Chlorella sp. contamination in a Phaeodactylum tricornutum culture
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SYSNO ASEP 0547567 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Comparison of various approaches to detect algal culture contamination: a case study of Chlorella sp. contamination in a Phaeodactylum tricornutum culture Author(s) Grivalský, Tomáš (MBU-M) ORCID
Střížek, Antonín (MBU-M)
Přibyl, Pavel (BU-J) RID, ORCID
Lukavský, Jaromír (BU-J) RID, ORCID
Čegan, Radim (BFU-R) RID, ORCID
Hobza, Roman (BFU-R) RID, ORCID
Hrouzek, Pavel (MBU-M) ORCIDSource Title Applied Microbiology and Biotechnology. - : Springer - ISSN 0175-7598
Roč. 105, č. 12 (2021), s. 5189-5200Number of pages 12 s. Language eng - English Country US - United States Keywords Phaeodactylum tricornutum ; Chlorella ; Microalgae ; Contamination detection ; Methods Subject RIV EE - Microbiology, Virology OECD category Microbiology Subject RIV - cooperation Institute of Botany - Microbiology, Virology
Institute of Biophysics - Biotechnology ; BionicsR&D Projects TN01000048 GA TA ČR - Technology Agency of the Czech Republic (TA ČR) Method of publishing Limited access Institutional support MBU-M - RVO:61388971 ; BU-J - RVO:67985939 ; BFU-R - RVO:68081707 UT WOS 000663488800001 EID SCOPUS 85108540559 DOI 10.1007/s00253-021-11396-7 Annotation Microalgal contamination in algal culture is a serious problem hampering the cultivation process, which can result in considerable economic and time losses. With the field of microalgal biotechnology on the rise, development of new tools for monitoring the cultures is of high importance. Here we present a case study of the detection of fast-growing green algae Chlorella vulgaris (as contaminant) in a diatom Phaeodactylum tricornutum culture using various approaches. We prepared mixed cultures of C. vulgaris and P. tricornutum in different cell-to-cell ratios in the range from 1:10(3) to 1:10(7). We compared the sensitivity among microscopy, cultivation-based technique, PCR, and qPCR. The detection of C. vulgaris contamination using light microscopy failed in samples containing cell ratios <1:10(5). Our results confirmed PCR/qPCR to provide the most reliable and sensitive results, with detection sensitivity close to 75 cells/mL. The method was similarly sensitive in a pure C. vulgaris culture as well as in a mixed culture containing 10(7)-times more P. tricornutum cells. A next-generation sequencing analysis revealed a positive discrimination of C. vulgaris during DNA extraction. The method of cultivation media exchange from sea water to fresh water, preferred by the Chlorella contaminant, demonstrated a presence of the contaminant with a sensitivity comparable to PCR approaches, albeit with a much longer detection time. The results suggest that a qPCR/PCR-based approach is the best choice for an early warning in the commercial culturing of microalgae. This method can be conveniently complemented with the substitution-cultivation method to test the proliferating potential of the contaminant. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2022 Electronic address https://link.springer.com/article/10.1007%2Fs00253-021-11396-7
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