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Evaluation of Bactericidal & Bacteriostatic

Invisible enemies of mankind are microorganisms. Despite being small, they have a profound effect on human bodies as well as other living organisms.
Invisible enemies of mankind are microorganisms. Despite being small, they have a profound effect on human bodies as well as other living organisms. Antimicrobial agents and drugs inhibit the multiplication of microbes or kill them. Antimicrobial drugs are found in nature as well as those discovered or developed.

However, because many pathogenic bacteria developed resistance to commonly prescribed antibiotics, it was necessary to look for other sources of antimicrobial substances, including plants. Many essential oils have been extracted from natural plants, including aromatics and medicinal. Essential oils are bioactive.

Material and Methods

Plant material

It is air-dried at room temperature and shaded for one week in the shade of the Taltemssen region (south of Agadir) from Thymus satureioides leaves and flowers harvested during May.

Microorganisms that are used and their growth conditions

Bacillus cereus, Acinetobacter baumannii, Escherichia coli, Staphylococcus aureus, Acinetobacter baumannii, and Staphylococcus aureus are the microorganisms used in this study. Several diseases are caused by the bacterial strains used in this study (urinary, intestinal, respiratory, etc.). These bacteria are grown on nutrient agar for 24 hours at 37°C in the dark.

Extraction of essential oils

Steam distillation was used to obtain the essential oils; 100 g of vegetable matter, accurately weighed, was mixed with 600 ml of distilled water and heated to boiling in a Clevenger apparatus. Sodium sulfate (Na2SO4) was used to dehydrate the oils and store them in dark bottles at 4 ° C to protect them from light and heat. Based on the dry vegetable matter, the essential oil yield is estimated.

Methods for evaluation

There are two methods in total

Diffusion disk method

A 24h bacterial suspension is seeded into Mueller Hinton in this method. With sterile tweezers, 6mm filter paper discs are deposited on the agar, previously impregnated with essential oils. Observing a clear halo around discs, if essential oils inhibited growth of the organisms over 24 hours, indicates that the essences inhibited growth of the organisms. A larger zone around a disk indicates a more sensitive germ.

Agar dilution method

There are five concentrations of essential oils prepared: a 10th, a 1/25, a 50th, a 100th, a 2nd, and a 500th diluted in a 0.2% agar solution. After autoclaving and cooling at 45 ° C, we add aseptically 1,5 ml of each of the dilutions to each test tube containing 13.5 ml of Mueller-Hinton reagent to obtain the final concentrations of 1/100, 1/250, 1/500, 1/1000, 1/2000, 1/3000 and 1/5000 (v/v). Before pouring the essential oils into Petri dishes, the tubes are properly agitated to disperse them. Cultures were also prepared in which the agar solution at 0.2% was added alone to the culture medium (Muller Hinton). An essential oil's minimum inhibitory concentration is the lowest concentration that inhibits visible growth after 16 to 20 hours of incubation at 37 ° C. A sample of Petri dish, on which there has been no growth of bacteria, is seeded with minimum bactericidal concentration (MBC). Upon incubation at 37°C for 24 hours, the minimum concentration of essential oils that would kill 99.99% of the bacteria corresponds to the MBC. Three treatments are supposed to be repeated three times to minimize the possibility of error.
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Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
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