Abstract
Objective: Melaleuca alternifolia (M. alternifolia) and its essential oil (EO) fractions have been used widely and traditionally in the treatment of various infectious diseases and hence its antibacterial potential is investigated in the present study.
 Methods: The antibacterial activity was studied through the agar disc diffusion method and broth dilution method, DNA fragmentation studies and confocal microscopy morphological studies were done. In-silico molecular interaction was studied against microbial target using docking software.
 Results: The inhibitory concentration of the EOs was recorded at 75% dilution with larger inhibition zones. The DNA fragmentation analyzed in the essential oil derived from in-vitro propagated leaves (EOIPL) of M. alternifolia treated bacterial cultures was compared with negative and positive controls. In Minimum Inhibitory Concentration (MIC) of EOIPL treated Staphylococcus aureus (S. aureus) showed time-dependent growth inhibition. The DNA content in the EOIPL treated bacterial cultures was comparatively less than in control cultures. The cell morphology changes of S. aureus cells were studied through confocal laser scanning microscopic analysis which showed a significant decrease in viable bacterial cells. The active component, terpinen-4-ol docked to autolysin receptor revealed stable interaction with the microbial target.
 Conclusion: Thus EOIPL was explored to possess bactericidal activity against common infectious bacteria and could in incorporated in therapeutic natural antibiotic formulations as with future studies.
Highlights
Since ancient times, medicinal plants have been used to maintain human healthcare [1]
The essential oil (EO) were extracted from all three sources of M. alternifolia through the steam distillation process and the oil content was comparatively analyzed
When compared with other bacteria, S. aureus was highly susceptible to essential oil derived from in-vitro propagated leaves (EOIPL) of M. alternifolia with larger inhibition zone whereas in E. coli showed less inhibition zone due to the presence of lipoproteins and lipopolysaccharides in their cellular walls that form a barrier to hydrophobic compounds
Summary
Medicinal plants have been used to maintain human healthcare [1]. WHO has recommended the development and use of environment-friendly alternative methods to control diseases [2]. Many plant extracts and essential oils isolated from plants have been shown to exert biological activity in-vitro and in-vivo, which justified research on traditional medicine focused on the characterization of antimicrobial activity of these plants [3]. The development of new antibiotics depends on the strategies such as bioavailability, targeting sites, route of administration, half-life period, spectrum of resistance. Melaleuca alternifolia (M. alternifolia) commonly known as Tea tree, M. alternifolia belongs to the family of Myrtaceae. Tea tree oil (TTO) of M. alternifolia contains various mono and sesquiterpenes, aromatic compounds. The capabilities of the unique and demonstrated anti-bacterial, anti-fungal, anti-viral, anti-inflammatory, and anti-septic, essential oil were first scientifically documented in 1905. The TTO is considered as a universal treatment for acne, eczema, and skin infections like herpes, warts, wounds, burns, nail mycosis, insect bites, colds, sore throat, gingival infections, haemorrhoids and vaginal infections [4]
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