National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
The global rise of antibiotic resistance against bacteria recent years increased the interest of research in alternative antimicrobial agents particularly those of the respiratory tract infection. Among these alternatives, Essential oils (EOs) are these mixtures of volatile compounds from plants more documented for their multiple therapeutic properties against airborne bacteria with multiple irreversible effects in-cell targets of bacteria. This study investigates the effect of EOs from Pentadiplandra brazzeana Baill. fresh roots, Allium sativum L. bulbs, Echinops giganteus A. Rich. dry roots on cell membrane integrity of Haemophilus influenzae by correlating their bactericidal action to cell release intracellular biomolecules. The essentials oils were extracted by hydrodistillation using a Clevenger apparatus. Then the inhibition parameters Minimal Inhibitory Concentrations (MIC) and Minimal Bactericidal Concentrations (MBC) of EOs and Chloramphenicol were investigated using a 96 wells microdilution assay against respiratory tract Gram- bacteria strains of Haemophilus influenza ATCC 49247, Pseudomonas aeruginosa ATCC 27853 and Klebsiella pneumoniae ATCC 17102. The dynamic action of each EOs on cell membrane integrity was demonstrated by showing their lethal effect on Haemophilus influenzae exposed at (MBC) of EOs, by quantifying the DNA release using the Nanodrop 1000 spectrophotometer and number of living cells at different times during 24 h at 37 °C. A correlation coefficient R2 involved between the DNA released and the number of living cells was established by plotting the curve concentration of DNA release versus the number of living cells counted in a solid medium. The results showed that, compared to Chloramphenicol, Pentadiplandra brazzeana and Allium sativum EOs exhibited strong activities with MICs of 9.76 μg/mL, and 39.06 μg/mL respectively, while E. giganteus EO showed moderate activity with MIC of 156.25 ng/µL. The lethal effect of Pentadiplandra brazzeana EO’s against H. influenza showed the decrease of living cells from 4.7 Log to 2 Log at the same time an increase of DNA concentration from 0 to 290 ng/µL with a symmetric axis of action at half time (280 min). Meanwhile, Allium sativum and Echinops giganteus EOs exhibited against H. influenza, a decrease of living cells from 4.7 Log to 2 Log and from 4.7 to 2.5 Log respectively and an increase of DNA concentration from 0 to 300 ng/µL after 420 min of exposition. In contrast, Chloramphenicol showed only lethal action against Haemophilus influenza, with a decrease of living cells from 4.7 Log to 0.2 Log and an increase of DNA concentration from 0 to 40 ng/µL at 500 min. Allium sativum and Echinops giganteus EOs correlated lethal and cell lysis effects respectively with R2 = 0.81 and R2 = 0.89 meanwhile Pentadiplandra brazzeana and Chloramphenicol correlated at 0.94 and 0.66 respectively. The action of EOs on Haemophilus influenzae cell membrane’s reveals a lethal effect directly correlated with cell lysis for Pentadiplandra brazzeana EO indirectly correlated with cell lysis for Allium sativum and Echinops giganteus EOs, depending on their antibacterial activity.
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