Formulation of Garlic Essential Oil-assisted Silver Nanoparticles and Mechanistic Evaluation of their Antimicrobial Activity against a Spectrum of Pathogenic Microorganisms.

BACKGROUND: The synthesis of nanoparticles using the principle of green chemistry has achieved huge potential in nanomedicine. Here, we report the synthesis of silver nanoparticles (Ag- NPs) employing garlic essential oil (GEO) due to wide applications of GEO in the biomedical and pharmaceutical industry. OBJECTIVE: This study aimed to synthesise garlic essential oil-assisted silver nanoparticles and present their antimicrobial and antibiofilm activities with mechanistic assessment. METHOD: Initially, the formulation of AgNPs was confirmed using different optical techniques, such as XRD, FT-IR, DLS, zeta potential, SEM, and EDX analysis, which confirmed the formulation of well-dispersed, stable, and spherical AgNPs. The antimicrobial and antibiofilm activity of GEO-assisted AgNPs was evaluated against a spectrum of pathogenic microorganisms, such as Gram-positive (S. aureus and B. subtilis) and Gram-negative (E. coli and P. aeruginosa) bacteria. RESULTS: The AgNPs exhibited remarkable antimicrobial and anti-biofilm activity against all tested strains. The mechanism behind the antimicrobial activity of AgNPs was explored by estimating the amount of reactive oxygen species (ROS) generated due to the interaction of AgNP with bacterial cells and observing the morphological changes of bacteria upon AgNP interaction. CONCLUSION: The findings of this study concluded that ROS generation due to the interaction of AgNPs with bacterial cells put stress on bacterial membranes, altering the morphology of bacteria, exhibiting remarkable antimicrobial activity, and preventing biofilm formation.

Mycobacterial biofilms: A therapeutic target against bacterial persistence and generation of antibiotic resistance.

Mycobacterium tuberculosis (M. tb) is the causative agent of Tuberculosis, one of the deadliest infectious diseases. According to the WHO Report 2023, in 2022, approximately 10.6 million people got infected with TB, and 1.6 million died. It has multiple antibiotics for treatment, but the major drawback of anti-tuberculosis therapy (ATT) is, its prolonged treatment duration. The major contributors to the lengthy treatment period are mycobacterial persistence and drug tolerance. Persistent M. tb is phenotypically drug tolerant and metabolically slow down which makes it difficult to be eliminated during ATT. These persisting bacteria are a huge reservoir of impending disease, waiting to get reactivated upon the onset of an immune compromising state. Directly Observed Treatment Short-course, although effective against replicating bacteria; fails to eliminate the drug-tolerant persisters making TB still the second-highest killer globally. There are different mechanisms for the development of drug-tolerant mycobacterial populations being investigated. Recently, the role of biofilms in the survival and host-evasion mechanism of persisters has come to light. Therefore, it is crucial to understand the mechanism of adaptation, survival and attainment of drug tolerance by persisting M. tb-populations, in order to design better immune responses and therapeutics for the effective elimination of these bacteria by reducing the duration of treatment and also circumvent the generation of drug-resistance to achieve the goal of global eradication of TB. This review summarizes the drug-tolerance mechanism and biofilms' role in providing a niche to dormant-M.tb. We also discuss methods of targeting biofilms to achieve sterile eradication of the mycobacteria and prevent its reactivation by achieving adequate immune responses.

Deciphering the Antibiofilm, Antibacterial, and Antioxidant Potential of Essential Oil from Indian Garlic and its Phytocompounds Against Foodborne Pathogens.

Garlic (Allium sativum L.), particularly its volatile essential oil, is widely recognized for medicinal properties. We have evaluated the efficacy of Indian Garlic Essential Oil (GEO) for antimicrobial and antibiofilm activity and its bioactive constituents. Allyl sulfur-rich compounds were identified as predominant phytochemicals in GEO, constituting 96.51% of total volatile oils, with 38% Diallyl trisulphide (DTS) as most abundant. GEO exhibited significant antibacterial activity against eleven bacteria, including three drug-resistant strains with minimum inhibitory concentrations (MICs) ranging from 78 to 1250 µg/mL. In bacterial growth kinetic assay GEO effectively inhibited growth of all tested strains at its ½ MIC. Antibiofilm activity was evident against two important human pathogens, S. aureus and P. aeruginosa. Mechanistic studies demonstrated that GEO disrupts bacterial cell membranes, leading to the release of nucleic acids, proteins, and reactive oxygen species. Additionally, GEO demonstrated potent antioxidant activity at IC50 31.18 mg/mL, while its isolated constituents, Diallyl disulphide (DDS) and Diallyl trisulphide (DTS), showed effective antibacterial activity ranging from 125 to 500 µg/mL and 250-1000 µg/mL respectively. Overall, GEO displayed promising antimicrobial and antibiofilm activity against enteric bacteria, suggesting its potential application in the food industry.

Antimycobacterial potential of Trachyspermum ammi seed essential oil via fume contact and determination of major compounds.

Trachyspermum ammi (L.), commonly known as carrom seeds or Ajwain, has been extensively studied for its medicinal properties. In this study, anti-mycobacterial effect of AEO in liquid and fume form was investigated against Mycobacterium smegmatis and Mycobacterium tuberculosis (M. tb). Results showed that AEO inhibits the growth of M. smegmatis at 0.03 mg/mL and becomes bactericidal at 0.125 mg/mL. MICs were observed at 0.03, 0.125 and 0.06 mg/mL against M. tb (H37Rv), isoniazid- and rifampicin-resistant (RIF-R) strains. Inverted disc-fume assay revealed AEO and Thymol efficiently inhibit the growth of M. smegmatis and M. tb. Similarly, in fume contact AEO and Thymol demonstrated antibiofilm activity at a dose of 1.25 mg/mL air and 40 mg/mL air against M.smegmatis effectively. GC-MS analysis showed that Thymol was the dominant compound. These findings suggest that the use of AEO in fume form may serve as a promising strategy as an anti-mycobacterial activity against M. tb.

GC/MS Profiling and Evaluation of Leaf Essential Oil for Bactericidal Effect and Free Radical Scavenging Activity of Plectranthus amboinicus (Lour.) Spreng Collected from Odisha, India.

Plectranthus amboinicus (Lour.) Spreng, known as the Indian borage or Mexican mint, is one of the most documented species in the family Lamiaceae for its therapeutic and pharmaceutical values. It is found in the tropical and subtropical regions of the world. The leaf essential oil has immense medicinal benefits like treating illnesses of the skin and disorders like colds, asthma, constipation, headaches, coughs, and fevers. After analyzing earlier reports with regard to the quantity and quality of leaf oil yield, we discovered that the germplasm taken from Odisha is preferable to other germplasms. The objective of the present work is to evaluate the free radical scavenging activity and bactericidal effect of leaf essential oil (EO) of Plectranthus amboinicus (Lour.) Spreng collected from the state of Odisha, India. The hydro distillation technique has been used for essential oil extraction. Upon GC/MS analysis, approximately 57 compounds were identified with Carvacrol as the major compound (peak area=20.25 %), followed by p-thymol (peak area=20.17 %), o-cymene (peak area=19.41 %) and carene (peak area=15.89 %). On evaluation of free radical scavenging activity, it was recorded that the best value of inhibitory concentration, was for DPPH with IC50 =18.64 ppm and for H2 O2 with IC50 =9.35 ppm. The EO showed efficient bactericidal effect against both gram positive (Mycobacterium smegmatis, Staphylococcus aureus, Enterococcus faecium) and gram negative (Escherichia coli, Vibrio cholerae, Klebsiella pneumoniae) bacteria studied through well diffusion method. Fumigatory action of the essential oil was found against M. smegmatis, the model organism for tuberculosis study. Alamar Blue assay, gave a result with MIC value for M. smegmatis i. e., 0.12 µg/ml and the MBC value of 0.12 µg/ml. Hence, P. amboinicus found in Odisha can be suggested as an elite variety and should be further investigated for efficient administration in drug formulation.