Abstract
Ocimum tenuiflorum (KT) is a common ethnobotanical plant of Southeast Asia. The ethnic communities of these regions use the various parts of the plants, especially the leaves, for the treatment of various ailments like cold and flu, chronic infections, and surface ailments. The leaves of these plants are consumed to act as immune boosters in the body. With this ethnical background, we performed the antimicrobial and antibiofilm potential of the methanolic extract of Ocimum tenuiflorum against biofilm formed by S. aureus biofilm. The biofilm formed by S. aureus is a potent cause for the development of gastrointestinal (GI)-associated chronic infection. The extract from the KT leaf was analyzed using UV spectroscopy and HPLC to confirm the presence of the active ingredients present within the extract. The HPLC and GC-MS studies revealed the presence of eugenol and linalool in a greater proportion having the maximum drug-like properties. It was observed that KT showed maximum inhibition of biofilms, proteins, and carbohydrates being present with the extracellular polymeric substance (EPS). Interestingly, the maximum inhibition to the quorum sensing (QS) and the genomic DNA, RNA content was reduced by eugenol and linalool in comparison to the plant extract. The studies were supported by in silico interaction between eugenol and linalool with the QS proteins of S. aureus. The studies were further confirmed with microscopic studies SEM and FCM. The IR studies also confirmed much reduction in biofilm when treated with eugenol, linalool, and KT with respect to the untreated sample.
Highlights
Microbial biofilm are complex association of sessile microorganisms attached to biotic or abiotic substrata with the help of extracellular polymeric substances (EPS) [1] forming a matrix and develops as a defense mechanism during extreme stressed conditions such as insufficient nutrient supply, extreme temperature, salt concentrations or pH variations etc
The major components found were gallic acid, p-coumaric acid, cinnamic acid, catechol, caffeic acid, 3,4dimethoxycinnamic acid, luteolin, diosmetin, kaempferol, apigenin, rosmarinic acid, genistein, eucalyptol, camphor and eugenol [28] Presence of various compounds eluted at different retention times were detected once the large compound fragments into small compounds resulting into characteristic peaks at different m/z ratios which can be further compared with the standard NIST library
Biofilms are formed by sessile microorganisms adhered to biotic/abiotic surfaces and are well protected within a self-secreted exopolymeric matrix consisting of proteins, carbohydrates, lipids, nucleic acids, small signaling molecules etc
Summary
Microbial biofilm are complex association of sessile microorganisms attached to biotic or abiotic substrata with the help of extracellular polymeric substances (EPS) [1] forming a matrix and develops as a defense mechanism during extreme stressed conditions such as insufficient nutrient supply, extreme temperature, salt concentrations or pH variations etc. The genetic diversity and inter-community interaction within the biofilm varies widely due to these stressed conditions forces the indwelling microbes to undergo a cell number density dependent sensing capabilities known as quorum sensing (QS) that helps in the survival of the microorganisms within the biofilm niche by modulating the biofilm formation via up/down regulating cascade of genes. QS or cell-cell communication is performed by small bacterial signalling molecules known as autoinducers (AI) that detects the neighboring population density and triggers specific bacterial genes controlling factors such as sporulation, antibiotic production, competence, virulence, motility and biofilm formation [2]. Though the signal transduction mechanisms, chemical nature of the signaling molecules and the target genes associated with various bacterial QS circuits differ widely in bacteria, ability of cell- cell communication and coordinated gene expression is a common factor in the bacterial society [3]
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