Black cardamom essential oil prevents Escherichia coli O157:H7 and Salmonella Typhimurium JSG 1748 biofilm formation through inhibition of quorum sensing

Abstract

This study aimed to investigate the chemical composition, using GC-MS, and anti-biofilm potential of black cardamom essential oil (BCEO) against biofilms of Escherichia coli O157:H7 and Salmonella Typhimurium JSG 1748 through inhibition of bacterial quorum sensing. GC-MS quantification demonstrated that BCEO contains 1,8-cineole (44.24%), α-terpinyl acetate (12.25%), nerolidol (6.03%), and sabinene (5.96%) as the major bioactive compounds. Antioxidant assays for BCEO revealed the total phenolic and flavonoid mean values were 1325.03 ± 7.69 mg GAE 100/g and 168.25 ± 5.26 mg CE/g, respectively. In regards to antimicrobial potential, Candida albicans was the most sensitive species compared to Streptococcus mutans, Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, and Salmonella Typhimurium with the following zones of inhibition; 14.4 ± 0.52, 13.2 ± 0.42, 11.2 ± 0.28, 11.0 ± 0.52, 8.2 ± 0.24 and 6.6 ± 0.18 mm in diameter, respectively. Biofilm inhibition by BCEO was concentration-dependent, when various concentrations of 0.03, 0.06, 0.12, 0.25 and 0.5% were applied, 33.67, 34.14, 38.66, 46.65 and 50.17% of Salmonella Typhimurium biofilm was inhibited, while 47.31, 54.15, 76.57, 83.36 and 84.63% of Escherichia coli biofilm formation was prevented. Chromobacterium violaceum ATCC 12,472 and its product violacein, was used as a microbial indicator for enhancement or inhibition of quorum sensing. Our data showed that 0.5% of BCEO inhibited violacein production without influencing the growth of Chromobacterium violaceum, while 1% of BCEO, caused 100% inhibtion of violacein production together with 30% inhibition of growth. This study shows that BCEO possesses promising antioxidant and antimicrobial potential, and found anti-biofilm activities linked to the quenching of the quorum sensing system of E. coli and S. Typhimurium.

Keywords: Anti-biofilm; Antioxidants; Black cardamom essential oil; GC–MS; Quorum sensing inhibition.

Fig. 1

Chromatogram indicating black cardamom essential oil components analyzed by Shimadzu GC 2010 Plus and GCMS-TQ8040 using Shimadzu SH-Rxi-5Sil MS column (30 m long, 0.25 mm ID, 0.25 μm film thickness). The area of the peak in the chromatogram reflect quantity (relative percentage, %) of each component present in the oil (for more details, see Table 1)

Fig. 2

Antimicrobial activity of black cardamom essential oil against planktonic and biofilm associated cells of E. coli O157:H7

Fig. 3

Antimicrobial activity of black cardamom essential oil against planktonic and biofilm associated cells of S. Typhimurium JSG 1748

Fig. 4

Quorum sensing inhibition (violacein production inhibition of C. violaceum ATCC 12,472) by black cardamom essential oil