ANTIBACTERIAL ACTIVITY FROM DIFFERENT PARTS OF MORINGA OLEIFERA (L.) AGAINST GRAM-POSITIVE AND GRAM-NEGATIVE BACTERIA

ANTIBACTERIAL ACTIVITY FROM DIFFERENT PARTS OF MORINGA OLEIFERA (L.) AGAINST GRAM-POSITIVE AND GRAM-NEGATIVE BACTERIA

Jitendra Kumar Verma, Tabish Qidwai * and Rohit Saluja

Faculty of Biotechnology, Institute of Bio-Sciences and Technology, Shri Ramswaroop Memorial University, Lucknow-Deva road, Barabanki, Uttar Pradesh, India.

ABSTRACT: Antibacterial activity of methanolic extracts of different parts (leaves, flowers, green pods and roots) of Moringa oleifera plant were evaluated against gram positive bacteria Bacillus subtilis (Bs), Staphylococcus aureus (Sa) and gram negative bacteria Klebsiella pneumonia (Kp), Escherichia coli (Ec), Pseudomonas aeruginosa (Pa). The activities were performed by the agar well diffusion and cup plate method, with methanol as the negative and tetracycline as a positive control. Secondary metabolites extracted from all the parts of M. oleifera were found active against bacteria. Furthermore, the antimicrobial activity of the mentioned plant extracts was also examined against various gram-positive and gram-negative bacterial strains. It was observed that against K. pneumonia, leaves extracts (17.6 mm) of M. oleifera possess higher antibacterial activity as compared to others, flower extract (19.2mm) against B. subtilis, leaves extracts (22.6 mm) more effective against E. coli, pod extracts (22.5 mm) against S. aureus and leaf extract (21.9 mm) showed against P. aeruginosa as compared to others. The results reveal that M. oleifera might be a potential source for the treatment of gram-positive and gram-negative bacteria.

Keywords: In-situ gel, Acyclovir, Anti-viral, HPMC E50 LV, Pluronic F-127

INTRODUCTION: Infectious diseases exert a significant socio-economic burden on countries, with a wide distribution of pathogens. Bacterial, plasmodium, and fungal infections affects people in many countries. The most acute group includes multidrug-resistant bacteria that pose a particular threat in hospitals, nursing homes, and among patients whose care requires devices such as ventilators and blood catheters. They cover Pseudomonas and various Enterobacteriaceae (including Klebsiella, E. coli, Serratia, and Proteus).

They can cause severe and often lethal infections ¹⁴. In addition, M. oleifera seeds shown strong antimicrobial effect against Enterococcus faecalis and Staphylococcus aureusstarins ^7-9. The seed extract and recombinant protein of M. oleifera are effective against Staphylococcus aureus, Streptococcus pyrogenes, Streptococcus mitis, Streptococcus pneumoniae, Enterococcus faecalis, Escherichia coli, and Legionella pneumophila for up to 150 min ¹².

Klebsiella pneumoniae are common intestinal bacteria that can cause lethal infections. Resistance in K. pneumoniae to last resort treatment (carbapenem antibiotics) has spread to all regions of the world. K. pneumoniae is a major cause of hospital-acquired infections such as pneumonia, bloodstream infections, and infections in newborn and intensive-care unit patients. In some countries, carbapenem antibiotics do not work in more than half of the patients treated for K. pneumoniae infections due to resistance (WHO, 2020). The development of drug resistance in pathogenic organisms has created problems in their treatments. Although a lot of research is being done effective treatment options are limited; therefore novel therapeutic compounds are urgently needed to prevent the aforementioned diseases.

Additionally, M. oleifera plant and its parts have well-known medicinal properties and have been used as an anti-inflammatory, anti-hypertensive, antimicrobial, anti-oxidant, anti-diabetic, anti-hyperlipidaemic, anti-neoplastic, anti-pyretic, anti-ulcer, cardio protectant and hepatoprotectant ¹⁰. Several important compounds, such as bactericidal and fungicidal, have been found in M. oleifera ^5-13. Methanolic extract of M. oleifera had a greater antibacterial effect than chlorhexidine (control group) ⁷. M. oleifera has gained fame as a nutritional immune booster in globally. Its consumption might be helpful in combination with traditional drugs like anti-malarial ¹¹. In the past, researchers focused only on leaves of M. oleifera for antibacterial properties. Current study aimed to investigate the antibacterial activities of methanolic extract of different parts of M. oleifera against disease-causing bacteria by disc agar diffusion method.

MATERIALS AND METHODS:

Collection of Plant Material: The leaves, flowers, green and pods of M. oleifera was collected from angelic region of Uttar Pradesh, India. Root collected from 30 days old plant in the nursery. The collected material was washed with water and shade dried, made to coarse powder and then packed in polythene bags for further analysis.

Extraction of Active Principles: The leaves, flowers, green pods, and root of M. oleifera has been dipped into polar and nonpolar solvents for the extraction. After 48 hours of incubation, filtration was done with polar & non-polar filtrates. The weighed amount of M. oleifera all parts powder was packed in an extraction thimble and placed in an extraction chamber which was suspended above the flask containing the solvent and below a condenser. In the last extraction process, the flask containing the extract was removed and solvent was evaporated using a rotary evaporator. The crude extract was stored in a refrigerator for antibacterial analysis.

Test Organisms: Gram positive bacteria B. subtilis (Bs), S. aureus (Sa), and gram negative bacteria K. pneumonia (Kp), E. coli (Ec), P. aeruginosa (Pa) were pure cultures used to test bacterial strains. The agar well diffusion and cup plate method performed the tests, with methanol as negative and tetracycline as a positive control.

Antibacterial Activity Assay: In-vitro antibacterial activity of selected plant extracts were tested by disc agar diffusion method ^4-15 was applied for antibacterial assay. The extracts were incorporated into appropriate medium and subsequently bacterial spore suspension was inoculated for bacterial susceptibility testing. The preparation was incubated at appropriate temperatures. After incubation, the zone of inhibition (diameter) originating in each disc of the medium was measured to resolve antibacterial activity of different parts of the plant. Each experiment was carried out in triplicate, and the mean diameter of the inhibition zone was measured in millimetre.

Determination of Minimum Inhibitory Concentration (MIC): The MIC for bacterial isolates was carried out using the tube dilution technique as described by Akinyemi et al. (2005) ¹. Whereas 0.5 mg of extracts and tetracycline was loaded from 5 mg / mL stock solutions from 100 µl of total volume.

RESULTS: Antibacterial activity of different parts of methanolic extract (leaves, flowers, green pods and root) of M. oleifera plant were analyzed against gram positive bacteria B. subtilis (Bs), S. aureus (Sa) and gram negative bacteria K. pneumonia (Kp), E. coli (Ec), P. aeruginosa (Pa). The present study suggested that antibacterial activity is shown in plant & their parts. In contrast, 0.5 mg concentration of both extracts and tetracycline was loaded from 5 mg / mL stock solutions from 100 µl of total volume. Secondary metabolites extracted from all four parts of M. oleifera were found to be active against bacteria. Furthermore, the antimicrobial activity of the mentioned plant extracts was also examined against various gram-positive and negative bacterial strains, and the results are summarized in Table 1. From the data, it was observed that against gram-negative bacteria K. pneumonia, leaves extracts (17.6 mm) of M. oleifera possess higher antimicrobial activity as compared to root extracts (16.1 mm), pod extracts (14.7mm) and lowest in flower extract (13.8 mm).

Antimicrobial activity of leaf extract (19.2mm) showed good antimicrobial activity against gram-positive bacteria B. subtilis as compared to pod extracts (14.3mm), root extracts (14.0 mm) and lowest in leaves extract (11.3 mm). However, leaves extracts (22.6mm) of the plant were found to be more effective against gram-negative bacteria E. coli than the pod extracts (19.6 mm), root extracts (16.9mm), and lowest in flower extracts (14.9 mm). Furthermore, pod extracts (22.5 mm) of the plant were found to be more effective against gram-positive bacteria S. aureus than the flower extracts (17.2 mm), root extracts (16.9 mm), and lowest in leaf extracts (14.1 mm).

Additionally, antimicrobial activity of leaf extract (21.9 mm) showed against gram-negative bacteria P. aeruginosa as compared to pod extracts (15.33 mm), but root and flower extracts no activity have been shown as described in Fig. 2, 3 and Table 1.

TABLE 1: ANTIBACTERIAL TESTING OF M. OLEIFERA, METHANOLIC EXTRACTION OF LEAF, ROOT, AND STEM AGAINST GRAM-POSITIVE BACTERIA B. SUBTILIS (BS), S. AUREUS (SA) AND GRAM-NEGATIVE BACTERIA K. PNEUMONIA (KP), E. COLI (EC), P. AERUGINOSA (PA)

Note: Value are presented as mean±S.E. of triplicate experiments.

FIG. 1: ANALYSIS OF ANTIBACTERIAL ACTIVITY OF DIFFERENT PARTS OF MORINGA OLEIFERA AGAINST GRAM-NEGATIVE AND GRAM-POSITIVE BACTERIA: 1-FLOWER EXTRACT, 2-POD EXTRACT, 3-ROOT EXTRACT, 4-LEAVES EXTRACT, 5-TETRACYCLINE AS CONTROL. A- ZONE OF INHIBITION (MM) OF B. SUBTILIS, B- ZONE OF INHIBITION (MM) OF S. AUREUS, C- ZONE OF INHIBITION (MM) OF K. PNEUMONIA, D- ZONE OF INHIBITION (MM) OF E. COLI, E-ZONE OF INHIBITION (MM) OF P. AERUGINOSA

After performing the antibacterial testing, it was found that extracts other than methanol could not show antibacterial activity as no clear zone of inhibition was obtained after the antibacterial sensitivity test.

FIG. 2: GRAPHICAL REPRESENTATION OF ANTIBACTERIAL ACTIVITY OF DIFFERENT PARTS OF M. OLEIFERA METHANOL EXTRACT AND TETRACYCLINE EFFECT AGAINST GRAM-POSITIVE BACTERIA.

FIG. 3: GRAPHICAL REPRESENTATION OF ANTIBACTERIAL ACTIVITY OF DIFFERENT PARTS OF M. OLEIFERA METHANOL EXTRACT AND TETRACYCLINE EFFECT AGAINST GRAM-NEGATIVE BACTERIA.

DISCUSSION: As the prevalence of diseases are being increased throughout the world, in which M. oleifera plant has rich in secondary metabolites expend therapeutic potential against a wide range of diseases, therefore, exploitation of M. oleifera and its products would be beneficial against the diseases above. In the present study four parts of the plant were used to evaluate for their antibacterial activity against gram positive bacteria B. subtilis (Bs), S. aureus (Sa) and gram negative bacteria K. pneumonia (Kp), E. coli (Ec), P. aeruginosa (Pa). The antimicrobial activities of M. oleiferaleaves, roots, bark and seeds were investigated in-vitro against bacteria, yeast, dermatophytes and helminths pathogenic to man ². Due to presence of glucosinolates and isothiocyanates; the stable isothiocyanate 4-[(α-l-rhamnosyloxy) benzyl] isothiocyanate (moringin) was evaluated for their antibacterial activity ⁶. Additionally, methanol, chloroform, ethyl acetate and aqueous bark extracts of Moringa oleifera were evaluated for their antibacterial activity against bacteria viz. Staphylococcus aureus, Citrobacter freundii, Bacillus megaterium and Pseudomonas fluorescens ¹⁵. Based on the results, it has been suggested that M. oleifera has antibacterial due to the presence of lipophilic compounds and different metabolites (carboxylic acid, 2, 4-diacetyl phloroglucinol, enzymes, and chitinases) in plant cell walls ⁸. Methanolic extract of flower and pod shown adequate activity against gram-positive bacteria B. subtilis (Bs), S. aureus( Sa) and leaves against gram-negative bacteria K. pneumonia (Kp), E. coli (Ec), P. aeruginosa (Pa).

CONCLUSION: The result of the present study showed that the methanolic extract of leaves and pod demonstrated better antibacterial activity against gram-positive and gram-negative bacteria than other plant parts. Leaves and pods can be used for further product development. Furthermore, it is important to evaluate key compounds responsible for the antibacterial activity.

ACKNOWLEDGEMENTS: The author thanks to Dr. Tabish Qidwai for providing innovative suggestions.

CONFLICTS OF INTEREST: The authors declare none conflict of interest statement.

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    How to cite this article:

    Verma JK, Qidwai T and Saluja R: Antibacterial activity from different parts of Moringa oleifera (L.) against gram-positive and gram-negative bacteria. Int J Pharm Sci & Res 2023; 14(6): 2894-98. doi: 10.13040/IJPSR.0975-8232.14(6).2894-98.

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