Microbial spoilage investigation of thawed common cuttlefish (Sepia officinalis) stored at 2 °C using next generation sequencing and volatilome analysis

Food Microbiol. 2018 Dec;76:518-525. doi: 10.1016/j.fm.2018.08.004. Epub 2018 Aug 6.


Cephalopods are highly appreciated with increasing demand seafood, but are also very perishable and deteriorate fast mainly due to microbiological spoilage. For this reason exploration of bacterial communities through 16S Next Generation Sequencing (NGS) and Volatile Organic Compounds (VOCs) analysis was performed. Furthermore, sensory evaluation, classical microbiological analysis, Total Volatile Base-Nitrogen/TVB-N and Trimethylamine-Nitrogen/TMA-N determination were also carried out. Shelf-life of thawed cuttlefish (Sepia officinalis) stored at 2°C determined by sensory evaluation was 4 days. Aerobic Plate Counts (APC) reached the levels of 6.6 log cfu/g. The initial and final population of all spoilage microorganisms enumerated with selective media was under detectable levels with the exception of Pseudomonas. Based on 16S NGS analysis, Psychrobacter were the dominants among others, e.g. Pseudomonas, Shewanella, Comamonas, Carnobacterium, Vagococcus, of the initial microbiota. Psychrobacter was also the dominant microorganisms of the spoiled cuttlefish. TVB-N and TMA-N increased considerably only at the late stages of storage. A plethora of VOCs were produced and some exhibited an increasing profile throughout storage, making them promising molecules as freshness indicators in contrast to TVB-N and TMA-N. The application of next generation sequencing revealed the microbiota that escapes the classic microbiological methodologies, showing that other microorganisms different from those determined on selective culture media might be the main cause of microbiological spoilage.

Keywords: 16S rRNA; Cephalopods; Cuttlefish; Next generation sequencing (NGS); Spoilage microbiota; Volatile organic compounds (VOCs).

MeSH terms

  • Animals
  • Bacteria / classification
  • Bacteria / genetics
  • Bacteria / growth & development*
  • Bacteria / isolation & purification*
  • Cold Temperature
  • Food Contamination / analysis
  • Food Storage
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Microbiota
  • Seafood / microbiology*
  • Sepia / microbiology*
  • Smell
  • Taste
  • Volatile Organic Compounds / chemistry*
  • Volatile Organic Compounds / metabolism


  • Volatile Organic Compounds