Annals of Microbiology

, Volume 69, Issue 3, pp 291–298 | Cite as

Comparative study of growth temperature impact on the susceptibility of biofilm-detached and planktonic Staphylococcus aureus cells to benzalkonium chloride

  • Simon Oussama Khelissa
  • Marwan Abdallah
  • Charafeddine Jama
  • Adem Gharsallaoui
  • Nour-Eddine ChihibEmail author
Original Article


The present study investigated and compared the effect of growth temperature on the susceptibility of biofilm-detached and planktonic Staphylococcus aureus cells, to benzalkonium chloride (BAC). This study also highlights the impact of BAC on the bacterial physiology and the role of membrane fluidity regulation as a bacterial resistance mechanism. The minimum inhibitory concentration of BAC was characterized with micro-dilution growth inhibition assay. The BAC treatment was performed on S. aureus cultured at 20 °C and 37 °C, for 24 h. The morphology of S. aureus cells was examined using scanning electron microscopy. The loss of bacterial membrane integrity after BAC treatment was studied by monitoring the intracellular potassium ion leakage using the atomic absorption spectroscopy. The bacterial membrane total fatty acid composition, controlling the membrane fluidity, was analyzed by GC/MS. The results showed that the resistance of S. aureus cells to BAC increased with the increase of growth temperature. The planktonic cells were more susceptible to BAC than biofilm-detached ones. The rise of growth temperature resulted in an increase of S. aureus membrane rigidity. Furthermore, a higher membrane fluidity was observed in planktonic cells when compared to that in the biofilm-detached ones. The resistance of S. aureus seems to depend on the growth temperature. Compared to planktonic cells, biofilm-detached cells showed a greater resistance to BAC. The BAC targets and disturbs the bacterial membrane. Membrane fluidity modulation is likely a one of resistance mechanisms for S. aureus to BAC at the cellular scale. Therefore, disinfection procedures, in food sector, should be adapted for bacteria detached from biofilm.


Staphylococcus aureus Biofilm-detached cells Planktonic cells Growth temperature Membrane fluidity Susceptibility to BAC 



The authors thank the Haut de France region and FEDER (Fonds européen de développement régional) for their financial support.


This work was carried out within the framework of ALIBIOTECH (Agroalimentaire et Biotechnologie, Nord Pas-de-Calais region) program.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent



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Copyright information

© Università degli studi di Milano 2018

Authors and Affiliations

  • Simon Oussama Khelissa
    • 1
  • Marwan Abdallah
    • 2
  • Charafeddine Jama
    • 2
  • Adem Gharsallaoui
    • 3
  • Nour-Eddine Chihib
    • 1
    Email author
  1. 1.CNRS, INRA, UMR 8207-UMET-PIHMUniversité de LilleVilleneuve d’AscqFrance
  2. 2.CNRS, ENSCL, UMR 8207-UMET-PSIUniversité de LilleVilleneuve d’AscqFrance
  3. 3.Université Lyon 1, ISARA Lyon, Laboratoire BioDyMIA, Equipe Mixte d’Accueil, no. 3733, IUT Lyon 1Technopole AlimentecBourg en BresseFrance

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