Annals of Microbiology

, Volume 63, Issue 2, pp 733–744

Characterisation and identification of spoilage psychotrophic Gram-negative bacteria originating from Tunisian fresh fish

  • Mouna Boulares
  • Mélika Mankai
  • Chedia Aouadhi
  • Ben Moussa olfa
  • Mnasser Hassouna
Original Article


The contamination of fresh fish by spoilage bacteria is undesirable, particularly when Gram-negative bacteria, which produce thermo-resistant protease and lipase, can grow. The purpose of the present work was, therefore, to isolate and identify psychrotrophic Gram-negative (Psy G(-)) bacteria, isolated from 80 samples of 12 species of wild and aquacultured fresh seafood, by biochemical and molecular methods using 16S rRNA gene sequencing. Twenty-eight identified strains were studied to evaluate their catalase, nitrate reductase, lipolytic and proteolytic activities, as well as growth ability, at different temperatures, pH and NaCl concentrations. Among 150 Psy G(-) strains, the most dominant species found were: Pseudomonas fluorescens, Aeromonas hydrophila, Pseudomonas putida and Photobacterium damselae. All strains of Psy G(-) had catalase activity and were able to reduce nitrates to nitrites. Proteolytic activity on milk and on gelatin agar was demonstrated for the majority of the isolates. However, extracellular proteolytic activity as assessed by the azocasein method wasn’t very high in all the strains. Lipolytic activity, as assessed by the agar method, showed that 92.9 % of strains could hydrolyse egg yolk, against 82.1 % and 57 % that could hydrolyse Tween 20 and Tween 80, respectively.


Fresh fish Psychrotrophic Gram-negative bacteria Proteolytic activity Lipolytic activity 


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Copyright information

© Springer-Verlag and the University of Milan 2012

Authors and Affiliations

  • Mouna Boulares
    • 1
  • Mélika Mankai
    • 1
  • Chedia Aouadhi
    • 2
  • Ben Moussa olfa
    • 1
  • Mnasser Hassouna
    • 1
  1. 1.Unité de Recherche “Sciences et Technologies des Aliments”, École Supérieure des Industries Alimentaires de Tunis (ESIAT)TunisTunisie
  2. 2.Laboratoire de Microbiologie, “Groupe bioprocédés”, Institut Pasteur de Tunis (IPT)TunisTunisie

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