ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES OF LEAF EXTRACTS OF BERBERIS ARISTATA AND ABIES WEBBIANA: THE ETHNOMEDICINAL PLANTS

ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES OF LEAF EXTRACTS OF BERBERIS ARISTATA AND ABIES WEBBIANA: THE ETHNOMEDICINAL PLANTS

Sucheta Gautam, Neetu Sachan, Avijit Mazumder, Rupa Mazumder and Dilipkumar Pal *

Department of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India.

ABSTRACT: This study was carried out to investigate the antibacterial and antifungal potentials of Abies webbiana and Berberis aristate Linn. The study aims to assess theantimicrobial activity and determine the zone of inhibition of extracts on some bacterial and fungal strains. The extracts determined the antimicrobial activity using the agar disc diffusion method. The antibacterial and antifungal activities of extracts (5, 25, 50, 100, 250 μg/ml, and so on) of A. webbiana and B. aristata were tested against different Gram-positive and Gram-negative bacteria and four fungal strains: Aspergillus niger, Candida albicans, and Penicillium notatum and Penicillium funiculus. The zone of inhibition of extracts was compared with that of a standard like ciprofloxacin for antibacterial activity and griseofulvin for antifungal activity. The results showed that the remarkable inhibition of the bacterial as well as fungal growth was shown against the tested organisms. The phytochemical analyses of the plant extracts indicated the presence of saponins, tannins, alkaloids, flavonoids, triterpenoids, steroids, glycosides, anthraquinones, coumarin, saponins, gum, protein, and amino acids. The antimicrobial activity of AWE and BAE may be due to the presence of such secondary metabolites. Hence, these plants may be used to discover bioactive natural products that may lead to the developing of new antimicrobials.

Keywords: Abies webbiana, Berberis aristate, Ethnomedicinal plants, Antibacterial and antifungal activities, Minimum inhibitory concentrations, zone of inhibition

INTRODUCTION: Antibiotics are one of our most important weapons in fighting bacterial infections and have greatly benefited the health-related quality of human life since their introduction.

However, over the past few decades, these health benefits are under threat. Many commonly used antibiotics have become less effective against certain illnesses because many produce toxic reactions and the emergence of drug-resistant bacteria.

It is essential to investigate newer drugs with lesser resistance. Drugs derived from natural sources play a significant role in preventing and treating human diseases. In many developing countries, traditional medicine is one of the primary healthcare systems ^1-13.

Herbs are widely exploited in traditional medicine. Their curative potentials are well documented ^14. About 61% of new drugs developed between 1981 and 2002 are based on natural products and have been very successful, especially in infectious disease and cancer ¹⁵. Recent trends, however, show that the discovery rate of active novel chemical entities is declining ¹⁶. Natural products of higher plants may give a new source of antimicrobial agents with possibly novel mechanisms of action ^17-18. The effects of plant extracts on bacteria have been studied by many researchers in different parts of the world ¹⁹. Much work has been done on ethnomedicinal plants in India ²⁰. Plants are rich in various secondary metabolites such as tannins, terpenoids, alkaloids, flavonoids, glycosides, etc., which have been found in-vitro to have antimicrobial properties ^21-22. Herbal medicines have been known to humans for centuries. Practitioners of traditional medicine ²³ have described the therapeutic efficacy of many indigenous plants for several disorders. Antimicrobial properties of medicinal plants are being increasingly reported from different parts of the world.

The World Health Organization estimates that plant extracts or their active constituents are used as folk medicine in traditional therapies of 80% of the world’s population ^24-28. The harmful microorganisms can be controlled with drugs. These results in the emergence of multiple drug-resistant bacteria and have created alarming clinical situations in treating infections. The pharmacological industries have produced several new antibiotics, and resistance to these drugs by microorganisms has increased gradually. Bacteria have the genetic ability to transmit and acquire resistance to synthetic drugs, which are utilized as therapeutic agents ²⁹.

To expand the spectrum of antibacterial agents from natural resources, Abies webbiana has been selected. In the Indian literature, this plant has been described to be useful against skin diseases, liver troubles, and tuberculosis glands, and its use in the treatment of hematemesis, fruits, leukoderma, and diabetes has been suggested. It has been concluded that plant parts could be used as a therapeutic agent in treating hypercholesterolemia partially due to their fiber and mucilage content.

Besides its pharmacological uses, the plant extract is also recommended as a pest and disease control agent in India. Berberis aristatais widely used by tribal people to treat various ailments, including ringworm and other fungal skin infections. The leaves are laxative, antiperiodic, depurative, anti-inflammatory and useful in skin diseases, boils, carbuncles, ulcers, intermittent fever, gouty arthritis and rheumatological disorders. A. webbiana and B. aristata plant organs are known to be an important source of secondary metabolites, they exhibit significant properties that support folkloric use in the treatment of some diseases caused by microbes. Thus, A. webbiana and B. aristata are well anchored in its traditional uses and now found wide-spread acceptance across the world ^29-31.

In the current investigation carried out, a screening of ethanol extracts of A. webbiana and B. aristata leaves against pathogenic bacteria and fungi is done to detect new sources of antimicrobial agents.

MATERIALS AND METHOD:

Collection of Plant Materials: The fresh and healthy leaves of the plant A. webbiana and B. aristate were collected between June and August 2016 from Kalka ji nursey Ranikhet (India). It is commonly known as Daruhaldi. The National botanical research institute, Lucknow, Uttar Pradesh (CIF-RB-3-207) authenticated the plant specimen. Plant parts were collected based on the information provided in the ethnobotanical survey of India. Each specimen/plant material was labelled and numbered and noted with the collection date, locality and medicinal uses recorded.

Preparation of Plant Extract:

Extraction: The leaves were dried in the shade at room temperature. The dried material was coarse powdered and packed in Soxhlet apparatus and extracted with petroleum ether (60-80 °C), ethyl acetate (70-75°C), and ethanol (78°C). The extraction of the A. webbiana and B. aristata leaves was carried out using known standard procedures ²⁸. The extracts were filtered using Whatman filter paper (No.1) while hot, concentrated in a vacuum under reduced pressure using a rotary flask evaporator, and dried in a desiccator. The alcoholic extract yields a dark greenish solid residue weighing 5.750 g (23.0% w/w). The ethanol extracts were then kept in sterile bottles, under refrigerated conditions, until further use. The extract was preserved at 2 to 4°C. These crude alcohol extracts (AWE and BAE, respectively) were used for further investigation for evaluation of antimicrobial properties.

Preliminary Phytochemical Screening: The extracts were subjected to preliminary phytochemical testing to detect the presence of different chemical groups of compounds for the presence of saponins, tannins, alkaloids, flavonoids, triterpenoids, steroids, glycosides, anthraquinones, coumarin, saponins, gum,  protein, and amino acids ^25-26.

Antimicrobial Activity: The antimicrobial activity of the plant extract was tested in-vitro against 21 microorganisms. Stock solutions (1 mg/mL) of both the alcohol extracts of the plants were prepared by dissolving each extract in dimethyl sulfoxide. Calculated volumes of stock solutions were dispensed in series of McCartney bottles previously containing calculated volumes of sterile cooled molten nutrient agar media (40–45°C) to prepare the volume of 30mL each with dilutions of 5, 10, 25, 50, 100,200, 400 and 800µg/mL these sterile nutrient agar media solutions were poured into Petri plates and allowed to solidify.

These plates were kept in the refrigerator at 4°C for 24h to allow the uniform diffusion of the compounds throughout the nutrient agar medium. Before spot inoculation, plates were kept at 37°C for 2 h. One loop full (loop diameter: 3 mm) of an overnight grown peptone-water culture of each microorganism was inoculated and the checkerboard technique marked the location of the inoculation. The spot inoculated plates were incubated at 37°C for 24h, and the minimum inhibitory concentration (MIC, mM) values were obtained ^25-37. The pMIC (−log10MIC) values of both the plant extract Berberis aristata (BAE) and Abies webbiana (AWE) were calculated. A MIC value of each extract was calculated and compared with that of the standard. Experiments were done in triplicate, and the results were presented as mean values of the three measurements.

Antifungal Activity: The antifungal activities of both the alcoholic extracts of the plants were studied by the disc diffusion assay method.

The pathogenic stains of different fungi given below in Tables 5-7 were used in the study for performing experiments.

RESULT AND DISCUSSION: Results of antibacterial studies are exhibited in Tables 1-7. From the results, it is found that the degree of effectiveness of microbial strain and the percent of inhibition were found to differ in both plant extracts at different concentration values. Both the alcoholic extracts of plants present in the research work showed inhibition against different bacterial and fungal MIC strains ranging from 50-200µg/ml.

In the case of antibacterial activity, the zone of inhibition of ethanol extract of B. aristate (BAE) ranged between 6.0 to 16.5 mm. At the same time, the ciprofloxacin showed a zone of inhibition between 16.0 to 20.5 mm. The extract showed lower antibacterial against Bacillus pumilus 82, whereas higher antibacterial activity was observed against Shigella flexneri Type 6.

The extract showed weak to moderate antibacterial activity against bacteria. The ethanol extract inhibited the growth of Staphylococcus aureus ML 267, Bacillus pumilus 82 and Bacillus subtilis ATCC 6633 at higher concentrations (200 µg/ml). BAE demonstrated maximum antibacterial activity here compared to AWE (Tables 1-4).

In the case of antifungal activity, the zone of inhibition of ethanol extract of A. webbiana (AWE) ranged between 12.5 to 14.5 mm, while the griseofulvin showed a zone of inhibition between 14.0 to 16.0 mm against fungal. The extract showed lower antifungal against Candida albicans ATCC 10231, whereas higher antifungal activity was observed against Penicillium notatum ATCC 11625. The extract showed weak to moderate antifungal activity against fungal strain. On the other hand, minimum inhibitory concentration values of BAE at 400 µg/ml were observed towards Aspergillus niger ATCC 6275 and correspond to the lowest concentration of extract inhibiting fungal growth. The ethanol extract inhibited the growth of C. albicans ATCC 10231 at higher concentrations (1000 µg/ml). Here, the AWE demonstrated maximum antifungal activity (Tables 5-7). The preliminary phytochemical screening found that AWE and BAE possess various phenolic and flavonoid contents. Although the antibacterial mechanisms of medicinal plants used in this study against various microorganisms were not fully illustrated, we suggest that phenolic and flavonoids may play an important role in their antibacterial activity. We suggest that plant extracts used in this study may become a source for discovering novel antibiotic agents from plant sources.

TABLE 1: DETERMINATION OF MINIMUM INHIBITORY CONCENTRATIONS OF THE SAMPLE CODED BAE

(0*-control (Sterile DMSO); + growth; - No growth; ± Inhibited growth)

TABLE 2: COMPARISON OF ZONES OF INHIBITION OF BAE WITH THAT OF STANDARD ANTIBACTERIAL AGENT CIPROFLOXACIN AT 200µG/ML

TABLE 3: DETERMINATION OF MINIMUM INHIBITORY CONCENTRATIONS OF THE SAMPLE CODED AWE

(0*-control (Sterile DMSO); + growth; - No growth; ± Inhibited growth)

TABLE 4: COMPARISON OF ZONES OF INHIBITION OF AWE WITH THAT OF STANDARD ANTIBACTERIAL AGENT CIPROFLOXACIN AT 200µG/ML

TABLE 5: DETERMINATION OF MINIMUM INHIBITORY CONCENTRATIONS OF THE SAMPLE CODED BAE

TABLE 6: DETERMINATION OF MINIMUM INHIBITORY CONCENTRATIONS OF THE SAMPLE CODED AWE

TABLE 7: COMPARISON OF ZONES OF INHIBITION OF VARIOUS SAMPLES AT 2000µg/ml

CONCLUSION: It is concluded that ethanol extract of B. aristate demonstrated maximum antibacterial activity compared to ethanol extract of A. webbiana. On the other hand, ethanol extract of A. webbiana Showed maximum antifungal activity compared to the ethanol extract of B. aristate. From the preliminary phytochemical screening, it was found that ethanol extract of A. webbiana and B. aristate possessed various phenolic and flavonoids contents. It is suggested that phenolic and flavonoids played an important role in their antibacterial activity.

ACKNOWLEDGEMENT: This work was supported by IFTM University Moradabad.

CONFLICTS OF INTEREST: The authors have no relevant conflicts of interest to declare.

REFERENCES:

1. Rani P, Pal D, Hegde RR and Hashim SR: Leuckart synthesis and pharmacological assessment of novel acetamide derivatives. Anticancer agent in Medicinal Chemistry 2016; 16(7): 898-906.
2. Rani P, Pal D, Hegde RR and Hashim SR: Acetamides: Chemotherapeutic agents for inflammation associated cancers. Journal of Chemotherapy 2016; 28(4): 255-65.
3. Saha S, Pal D and Kumar S: Antifungal and antibacterial activities of phenyl and ortho hydroxyl phenyl linked imidazolyl triazolo hydroxamic acid derivatives. Inventi Rapid Med Chem 2017; 1: 1-8.
4. Goutam S, Sachan N, Shrivastav A and Pal D: Evaluation of antidepressant activity of ethanolic extract of Abies webbiana and Berberis aristata in Laboratory Animals. J of Drug Delivery and Therapeutics 2019; 9(1): 244-247.
5. Rani P, Pal D, Hegre RR and Hashim SR: New mercaptoacetamide derivatives: synthesis and assessment as antimicrobial and antimycobacterial agents. Pharmaceutical Chemistry Journal 2021; 55 (7): 715-23.
6. Saccan N, Chandra P, Pal D: Assessment of gastroprotective potential of Dalonixrevia (Boj-Ex Sooa) Raf. against ethanol and cold rextrin stressed-induced ulcer in rats. Tropical J of Pharma Res 2015; 14(6): 1063-70.
7. Kanta C, Sharma IP and Sheikh MA: Ethnobotanical studies on medicinal plants of Longata area, Kupwara, Jammu and Kashmir, Indian Journal of Medicinal Plants Studies 2018; 6(2): 94-7.
8. Yelmeta AK and Thonte SS: Antimicrobial activity and FTIR spectroscopic analysis of Argemona mexicana L leaves extracts, International Journal of Pharmaceutical Sciences and Research 2021, 12 (12): 6653-59.
9. Jaykant Vora A, Anshu Srivastava B and Hashmukh Modi A: Antibacterial and antioxidant strategies for acne treatment through plant extracts. Informatics in Medicine Unlocked 2019; 2(13): 128-32.
10. Suresh KD, Choudhury PK, Srivastava R and Sharma M: Antimicrobial, anti-inflammatory and wound healing activity of polyherbal formulations. Biomedicine and Pharmacotherapy 2019; 2(1): 555-67.
11. Sindhu RK, Chitkara M. Gagandeep K, Kaur A, Arora S, IS: Sandhu: Formulation development and antimicrobial evaluation of polyherbal soap. Plant Archieves 2019; 2: 1342-46.
12. Hintz T, Matthews KK and Di R: The use of plant antimicrobial compounds for food preservation. Bio Med Research International 2015; 2015: Article ID 246264.
13. Lucera A, Costa C, Conte A and Del Nobile MA: Food applications of natural antimicrobial compounds Frontiers in Microbiology 2012; 3: 287.
14. Lee KW and Lip GYH: The role of omega-3 fatty acids inthe secondary prevention of cardiovascular disease. QJM 2003; 96: 465–80.
15. Wu HS, Raza W, Fan JQ, Sun YG, Bao W and Shen QR: Cinnamic acid inhibits growth but stimulates production of pathogenesis factors by in vitro cultures of Fusarium oxysporum sp. niveum. JAFC 2008; 56: 1316.
16. BagchiK and Puri S: Free radicals and antioxidants in healthand disease. EMHJ 1998; 4: 350-60.
17. Kim YW and Byzova TV: Oxidative stress in angiogenesis and vascular disease. Blood 2014; 123: 625-631.
18. Young IS and Woodside JV: Antioxidants in health and disease. Journal of Clinical Pathology 2001; 54: 176-86.
19. Pal D and Nimse SB: Screening of the antioxidant activity of verticillata plant. Asian J of Chem 2006; 18: 3004-8.
20. Suh HJ, Chung MS and Cho YH: Estimated daily intakes of butylate hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl hydroquinone (TBHQ) antioxidants in Korea. Food Additives and Contaminants 2005; 22: 1176–1188.
21. Lafay S and Gil-Izquierdo A: Bioavailability of phenolic acids. Phytochemistry Reviews 2008; 7: 301-11.
22. Clifford MN: Chlorogenic acids and other cinnamates nature, occurrence, dietary burden, absorption and metabolism. J of the Sci of Fo and Agri 2000; 80: 1033-43.
23. De P, Baltas M and Bedos-Belval F: Cinnamic acid derivativesas anticancer agents-a review. Current Medicinal Chemistry 2011; 18: 1672-1703.
24. Pal D, Mandal M, Senthilkumar GP and Padhiari A: Antibacterial activity of methanol extract of Cuscuta reflexa stem and Corchorus olitorius Linn. Seed. Fitoterapia 2006; 77: 589-91.
25. Mohanta TK, Patra JK, Rath SK, Pal D and Thatoi HN: Evaluation of antimicrobial activity and phytochemical screening of oils and nuts of Semicarpusana cardium f., Scientific Research and Essays 2007; 2: 486- 490.
26. Pal D, Tripathy R, Pandey DD and Mishra P: Synthesis, characterization, antimicrobial and pharmacological evaluation of some 2,5-disubstituted sulphonyl amino 1,3,4- oxadiazole and 2-amino-disubstituted 1,3,4-thiadiazole derivatives. Journal of Advanced Pharmaceutical Technology and Research 2014; 196-201.
27. Nimse SB, Pal D, Mazumder A and Mazumder R: Anti-oxidant and antimicrobial activity of amide derivatives of cinnamic acid. Journal of Chemistry 2015.
28. Gurjar VK, Pal D, Mazumdar A and Mazumdar R: Synthesis, Biological Evaluation and Molecular Docking studies of novel 1,8 Naphthyridine-3-carboxylic acid derivatives as potential antimicrobial agents (Part-1). Indian J of Pharmaceutical Sciences 2020; 82: 41-53.
29. Sova M: Antioxidant and antimicrobial activities of cinnamicacid derivatives, Mini- Reviews in Medicinal Chemistry 2012; 12: 749-67.
30. Vishnoi S, Agrawal V and Kasana VK: Synthesisand structure activity relationships of substituted cinnamic acidsand amide analogues: a new class of herbicides; Journal of Agricultural and Food Chemistry 2009; 57: 326.
31. S´anchez-Moreno C: Review: methods used to evaluate thefree radical scavenging activity in foods and biological systems. Food Science and Tech Inter 2002; 8(3): 121–37.
32. Rani P, Pal D, Hegde RR and Hashim SR: Synthesis, characterization and pharmacological evaluation of substituted phenoxy acetamide derivatives. Hemijskaindustrija 2015; 69(4):405-15.
33. S´anchez-Moreno C, Larrauri JA and Saura-Calixto F: Aprocedure tomeasure the antiradical efficiency of polyphenols. Journal of the Science of Food and Agriculture 1998; 76(2): 270–6.
34. Oktay M, Gulcin I and Kufrevioglu OI: Determination of in-vitro antioxidant activity of fennel (Foeniculum vulgare) seed extracts. LWT Food Science and Technology 2003; 36(2): 263–71.
35. Nimse SB and Pal D: Free radicals, natural antioxidants, and their reaction mechanisms. RSC Advances 2015; 5(35): 27986–28006.
36. Ali HM, Abo-Shady A and Eldeen HAS: Structural features, kinetics and SAR study of radical scavenging and antioxidant activities of phenolic and aniline compounds. Chemistry Central Journal 2013; 7(1): 53-61.
37. Guna G: Antimicrobial activity of some ethnomedicinal plants used in Kashmir. India. International J of Pharma Sciences and Research 2018; 9(2): 5339-43.
    

    ---

    How to cite this article:

    Gautam S, Sachan N, Mazumder A, Mazumder R and Pal D: Antibacterial and antifungal activities of leaf extracts of Berberis aristata and Abies webbiana: the ethnomedicinal plants. Int J Pharm Sci & Res 2022; 13(8): 3350-56. doi: 10.13040/IJPSR.0975-8232.13(8).3350-56.

    ---

    All © 2022 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.