Related Experiment Video
Updated: Jun 17, 2025

Antimicrobial Synergy Testing by the Inkjet Printer-assisted Automated Checkerboard Array and the Manual Time-kill Method
Published on: April 18, 2019
Synergistic interaction between Chlorella vulgaris extract and Origanum elongatum essential oil against
Sanaa Ouzakar1, Nadia Skali Senhaji2, Abdeltif El Harsal2
1Biotechnology and Applied Microbiology Team, Research Laboratory of Applied Chemistry and Microbiology and Biotechnology, Department of Biology, Faculty of Sciences, Abdelmalek Essaadi University, Tetouan, Morocco. sanaeouzakar@gmail.com.
Abstract:
The massive emergence of antimicrobial resistance in recent decades has rendered the use of a single-agent strategy ineffective. Consequently, the combination of different therapeutic agents has emerged as a promising new approach. The aim of the present study was to investigate the combined effect of Chlorella vulgaris methanol extract (CVME) and Origanum elongatum essential oil (OEEO) on methicillin-resistant Staphylococcus aureus (MRSA). Thus, the antibacterial activity of OEEO and CVME on Escherichia coli, Staphylococcus aureus, and MRSA was evaluated using the agar well diffusion and broth microdilution methods. The killing activity of CVME and OEEO, individually and in combination, on MRSA ATCC 43300 was tested using the time-kill assay. The synergistic effect was examined by determining the fractional inhibitory concentration index (FICI) using the checkerboard test. The results showed very significant antibacterial activity against all the bacteria tested, for both OEEO and CVME, with minimum inhibitory concentrations (MICs) ranging from 0.125 to 0.25% (v/v) for OEEO and from 3.12 to 6.25 mg mL-1 for CVME. Minimum bactericidal concentration (MBC) values for OEEO and CVME were in the range 0.125-0.5% (v/v) and 6.25-12.5 mg mL-1, respectively. The inhibition zones associated with OEEO were distinctly greater than those associated with CVME for all the bacteria examined. When used individually, the time-kill curves of OEEO and CVME revealed a dose-dependent effect on MRSA proliferation. Compared with controls, both agents were able to prolong the latent phase of growth curves and decelerate bacterial growth. The killing effect of OEEO on MRSA was considerably higher than that observed with CVME. OEEO prevented MRSA proliferation at only 1/2 of its MIC, while the CVME did so at 2 times its MIC. The combination of OEEO with CVME demonstrated a synergistic effect against MRSA, with a FIC index value of 0.49. The findings therefore suggest that the combination of C. vulgaris methanol extract and O. elongatum essential oil at very low doses may be promising anti-MRSA candidates. A search of the published literature revealed that, to our knowledge, no studies have yet been carried out on the antibacterial potential of combining essential oils and microalgae extracts in the fight against MRSA.
Related Concept Videos
Combined Effects of Drugs: Synergism
Such synergistic combinations...
Hand hygiene
Hand washing...
Surface Membrane Barriers
The outer layer of the skin, the epidermis, is a robust barrier comprising layers of closely packed keratinized cells. This dense arrangement prevents microbes from penetrating the body. The periodic shedding of epidermal cells...

