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Updated: Jun 1, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Seafood quality analysis: Molecular identification of dominant microbiota after ice storage on several general growth
K Broekaert1, M Heyndrickx, L Herman
1Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science, Brusselsesteenweg 370, 9090 Melle, Belgium. Katrien.Broekaert@ilvo.vlaanderen.be
Abstract:
This study points out the limitations of several general growth media frequently used in seafood research by a systematic identification of the microorganisms on fish samples during ice storage unable to grow on those media. Aerobic psychrotrophic count (APC), replication on various general media and total cultivable microbial community denaturing gradient gel electrophoresis (DGGE) analysis revealed that many potential spoilage microorganisms were overlooked. Those microorganisms overlooked by using only one single growth medium were identified by partial 16S rRNA gene and gyrB gene sequencing. Members of the genera Shewanella, Vibrio, Aliivibrio, Photobacterium, Pseudoalteromonas and Psychrobacter, including Photobacterium phosphoreum, Shewanella baltica and Pseudomonas fluorescens are unable to grow on PCA. APC analysis also confirmed that on plate count agar (PCA) the enumeration of the microbiota was underestimated. Although Long and Hammer agar (LH) and marine agar (MA) obtained the best quantitative (APC analysis) and qualitative (replication and DGGE analyses) results for fish quality analysis, analysts have to keep in mind that some species were also unable to grow on those media, such as Pseudomonas fragi and Acinetobacter sp.
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