Chemical characterization of Thymus zygis L. (thyme) and Elettaria cardamomum L. (cardamom) essential oils and investigation of their inhibitory potentials against cervicovaginal Escherichia coli isolates

Abstract

Introduction

The cervicovaginal microbiota plays a crucial role in women's well-being and reproductive health. This study initially focuses on identifying the chemical components of the essential oils (EOs) from Elettaria cardamomum L. (cardamom) (Ec) and Thymus zygis L. (thyme) (Tz). Subsequently, this study aims to determine the antimicrobial and biofilm inhibitory effects of these EOs against both cervicovaginal mixed cultures and the microbial strains isolated from them.

Methods

The compositions of EOs were determined by GC–MS analysis using an Agilent 7890B GC/5977A Series MSD System. Cervicovaginal samples were obtained from two women presenting complaints of a strong odor and vaginal discharge. Samples were collected from the cervix using a sterile cytobrush, ensuring no contact with the urethra, rectum, or external parts of the vagina. The cytological examination was conducted using Papanicolaou staining. Isolated microorganisms were identified through 16S Ribosomal RNA Gene Sequence Analysis and characterized using antibiogram tests. Antimicrobial tests of the EOs were performed by broth microdilution and disc diffusion methods. Biofilm formation was assessed using a crystal violet binding assay.

Results

The major components of Ec EO were α-terpinyl acetate (41.95 %) and eucalyptol (28.58 %), whereas Tz EO contained significant amounts of o-cymene (38.76 %), carvacrol (22.66 %), and thymol (20.69 %). Escherichia coli codes OR651248 and OR651249 were isolated from CV 6 and CV 7 mixed cultures, respectively. E. coli OR651248 demonstrated a higher antibiotic resistance level than E. coli OR651249. Biofilm formation by mixed cultures was significantly higher than that by E. coli isolates (P < 0.05). The Ec EO displayed minimal to no effect against both mixed cultures, showing zero inhibition zone, 500 μL/mL minimum inhibitory concentrations (MICs), and 500 μL/mL minimum microbicidal concentrations (MMCs). However, the Tz EO exhibited antimicrobial activity against both mixed cultures and the E. coli isolates, with inhibition zones > 15 mm and MICs/MMCs < 4 μL/mL (P < 0.05). In all microbial cultures, 0.5 MIC of both EOs inhibited biofilm formation by more than 80 %.

Conclusion

The Tz EO exhibits promising potential as an antimicrobial and biofilm inhibitory agent for managing cervicovaginal E. coli infections.