ANTIMICROBIAL ACTIVITY OF DIFFERENT VARIETIES OF TERMINALIA CATAPPA LEAVES

ANTIMICROBIAL ACTIVITY OF DIFFERENT VARIETIES OF TERMINALIA CATAPPA LEAVES

M. I. Farzana Tasneem ^{* 1} and P. N. Narsegowda ²

Tumkur University ¹, Tumkur - 572101, Karnataka, India.

Sangenomics Research Labs ², Hoskote, Bengaluru - 562114, Karnataka, India.

ABSTRACT: Terminalia catappa Linn. belongs to the family Combretraceae found in tropical regions of Asia, Africa and Australia, which is commonly called as Indian almond, sea-almond, tropical almond, wild badam. The fruit is useful in bronchitis and bowels. Juice of leaves is used in the preparation of the ointment for scabies, leprosy, cutaneous diseases and also as analgesic. The root bark is used in dysentery and diarrhoea. The stem bark cures fevers. The oil yielded from kernels is used commercially as a substituted for almond oil. According to the literature the different parts of the plants is used in many therapies including, antibacterial anticlastogenic, antioxidant, antifungal, antiulcer and molluscidal activity, management of sickle cell aneamia, anticancer, anti HIV reverse transcriptase, hepato- protective, anti-inflammatory, antidiabetic. In this study the methanol extracts of two varieties of Terminalia catappa (yellow and red variety) leaf was screened for antifungal like Candida albicans and Aspergillus niger and antibacterial like Eschiershea coli, Staphylococcus aureus, Citrobacter species, Enterococcus faecalis, Pseudomonas aerogenosa  and Bacillus  cereus  was evaluated. The variation in the intensity of inhibitory activity of this microorganism between two varieties was observed.

Terminalia catappa, Antibcaterial, Antifungal, Staphylococcus  aureus, Candida albicans

INTRODUCTION: Herbal medicines have been known to man for centuries, the therapeutic efficacy of many indigenous plants for several disorders discovered by well known scientist. Antimicrobial properties of medicinal plants are being increased reported from different parts of the world, the plant extract or their active constituents used as fold medicines in traditional therapies of 80% of the world’s population.

The harmful microorganism can be controlled with the drugs and these results in the emergence of multiple drug resistant bacteria and it has created an alarming clinical situation in the treatment of infections, in effort to expand the spectrum of antibacterial agents from natural resources.

Medicinal plants contain active chemical constituent in various parts of plants like root, leaf and stem. Plant extract has active ingredients ¹. The combination of plant extract and antibiotics could be useful in fighting emerging drug - resistance microorganisms. Terminalia catappa Linn. is an ornamental tropical tree belonging to the family Combretaceae is native to Southeast Asia in the tropical regions of Asia, Africa and Australia. It is known by the common names Bengal almond, Country almond, false kamani, Indian almond, Malabar almond, and about 35m (115 ft) tall, 9 m in width of its symmetrical canopy ² and horizontal branches. The greenish yellow leaves are clustered in axillary spikes, small and inconspicuous. It usually commences flowering within 2 - 3 years of out planting but this may vary with site and genotype. It is a large deciduous stately tree with a characteristic pagoda shape ³. It is cultivated in Nigeria solely as a shade tree and for its fruits and seeds as well as medicinal uses ⁴. The several reports revealed the identification, characterization and standardization of Terminalia catappa as medicinally important plant ^{5, 6}.

Plant with red leaves are used to expel worms applied to rheumatism where as ordinary leaves are mixed with the oil and rubbed on to the breast to relieve mammary pain, secondary metabolites which are useful for the growth and development of plant ⁷. The pharmacological significance of the plant is an updated review is for the traditional medicinal uses which play a major chemical constitutes reported on this tree and plant was sensitive to pollution control in Nature ⁸.

Many scientists have contributed and studied phytochemicals ⁹ play an important role in the treatment of different types of diseases and disorders and are still used in both traditional and modern medicine. Phenolic compounds are known as powerful chain breaking antioxidants ¹⁰. Juices from leaves were used as an ointment for scabies, leprosy and other skin infections ¹¹. The plant showed a rich source of bioactive compounds and justifying their traditional use to treat any disease and disorder is postulated. The leaves have antioxidant property anticancer used to cure liver diseases ¹² anti HIV reverse transcriptase and management of sickle cell anemia ¹³, hepato-protective, anti-inflammatory ¹⁴ antidiabetic  ^{15, 16} fish pathogen ¹⁷.

The leaves found to possess to fight against the infectious microbes which might be due to the phytochemicals identified in the leaf thus acting as a potent antimicrobial agent ^{18, 19, 20, 21, 22}. Methanol extract showed significant antifungal activity against most susceptible mold ^{23, 24}. Terminalia catappa as a source of naturally derived anti-microbial formulations to herbal medicines against bacterial infection ²⁵ and in pharma industry. In present study the methanol extracts of two varieties of Terminalia catappa (yellow and red variety) leaf was screened for antimicrobial activities.

MATERIALS AND METHODS:

Collection of Material: The leaves of Terminalia catappa were collected in the reference of two varieties that is the yellow and red variety from Bangalore city. The two yellow varieties are called as T1 and T2. The two red varieties were called as T3 and T4.

Preparation of Plant Extract:

Extraction: The leaves of Terminalia catappa were air dried and then the sample was powdered in a homogenizer 10 g was used for methanol extraction by Soxhlet method. The solvent was then evaporated to dryness under reduced pressure in Roto evaporator and the extracted compound left was used for the antimicrobial assay.

Preliminary Phytochemical Assay: The methanolic extract of different varieties of Terminalia catappa leaves were subjected to preliminary phyto-chemical assay  to detect the presence of different bioactive compounds like proteins, carbohydrates, glycosides, flavanoids, tannins and phenol ^{26, 27}.

Quantitative Analysis of Phytochemicals:

Determination of Total Phenol: The total phenol concentration in methanolic leaf extracts were determined by using Folin-Ciocalteu method ²⁸. To 1 ml of leaf extract, 0.5 ml of 0.2N Folin- Ciocalteu reagent and 1.5 ml of 0.7 M sodium carbonate solution was added and adjusted to 10ml with distilled water. After incubation in dark at 25 ºC for 1 h, the absorbency of reaction mixture was measured at 725 nm. Catechol was used as a standard to produce the calibration curve.

Determination of Total Flavonoids: The total flavonoid content of leaf extract was measured using aluminium chloride method ²⁹. One ml of leaf extract was mixed with 0.5 ml of 1.2% aluminium chloride and 0.5 ml of 120 mM potassium acetate. The volume was then increased to 10 ml with distilled water. The mixture was incubated for 30 min at room temperature. The absorbance was measured at 415 nm. Rutin was used as standard for the calibration curve.

Test Microorganisms and Growth Media: The following microorganisms such as Eschiershea coli, Pseudomonas aerogenosa, Bacillus cereus, Staphylococcus aureus, Staphylococcus mutans, Enterococcus and fungal strains like Candida albicans, A. niger, Enterococcus, Klebsiella, Citrobacter were chosen based on their clinical and pharmacological importance.

The bacterial and fungal stock strains were incubated for 24 h at 37 ºC on nutrient agar and potato dextrose agar medium (PDA), stock cultures were maintained at 4 ºC. All bacterial strains were procured in lyophilized form from the Institute of Microbial Technology (IMTECH), Chandigarh.

Antimicrobial Activity:

Determination of Zone of Inhibition: In-vitro antibacterial and antifungal activities were examined for methanol extract against six pathogenic bacteria (Gram positive and Gram negative) and two pathogenic fungal were investigated by well diffusion method. Each purified extracts were dissolved in methanol for the determination of zone of inhibition and standard antibiotic ampicillin (10 mg/ml) used for comparison of the results.

All the extracts (T1, T2, T3, T4) were screened with different concentration of plant extract (50, 100, 150, 200 μg/ml) against the Eschiershea coli, Pseudomonas aerogenosa, Bacillus cereus, S. aureus, Staphylococcus mutans, Enterococcus, Klebsiella pneuomonia, Citrobacter and fungal strains like Candida albicans, Aspergillus niger. The zones of growth inhibition around the discs were measured after 18 to 24 h in incubation at 37ºC and 28 ºC for bacterial and fungal respectively. For each extract three replicate trials were conducted against each organism. The diameters of the inhibition zones were measured in mm.

RESULTS AND DISCUSSION:  According to the result of qualitative analysis of chemical constitutes of different varieties in Terminalia catappa  showed the presence of amino acid, protein, carbohydrates, glycosides, alkaloids (+) and  saponin, tannin, flavanoid showed very good result (++) shown in Table 1.

The quantitative analysis with a focus on flavanoids, and phenol from the methanol extract of different varieties of the sample in Table 2. The phenol content was determined by Folin-ciocalteau reagent. The maximum concentration of phenol was observed T3-195 µg/ml, T4-185 µg/ml, T2- 180 µg/ml minimum concentration was observed in-T1-178 µg/ml. The flavanoid content was evaluated by aluminium chloride method highest amount of flavanoid content was obtained in sample T1-0.89 µg/ml, T4-0.86 µg/ml, T3-0.75 µg/ml and low in sample T2-0.72 µg/ml.

TABLE 1: QUALITATIVE ANALYSIS OF LEAF EXTRACT

TABLE 2:  QUANTITATIVE ANALYSIS OF PHENOL AND FLAVANOID

FIG. 1: REPRESENTING THE QUANTITATIVE ESTIMATION OF PHENOL AND FLAVANOID

Antimicrobial activity of methanol extract of Terminalia catappa leaf variety was studied with selected G+ve and G-ve pathogens such as Bacillus cereus, Staphylococcus aureus, K. pneumonea, Eschiershea coli, Citrobacter sp., Pseudomonas aerogenosa and fungus such as Candida albicans and Aspergillus niger was performed in well diffusion method.

Result revealed that the methanol extract of Terminalia catappa leaves was found to be effective against standard ampicillin (40 mm), G +ve such as Staphylococcus aureus   with the zone of inhibition is 40 mm in T1 and T2, 30 mm in T3, 20 mm in T4. The G+ve Bacillus cereus showed the zone of inhibition 22 mm in T2 and T3, no zone of inhibition in T1 and T4.

An antifungal activity of G+ve like Candida albicans showed 12 mm in T1, 26 mm in T2, 07 mm in T4, no zone of inhibition in T3. Aspergillus niger with the zone of inhibition as 20 mm in T2,18 mm in T3, 10 mm in T4, no zone of inhibition in T1. G–ve bacteria like Eschiershea coli showed 40 mm in T1, T2 and 32 mm in T3, 20 mm in T4. Pseudomonas aeurogenosa has 20 mm in T1 and 22mm in T2 and 19 mm in T3, 11mm in T4. Citrobacter sp showed 16 mm in T1, 12 mm in T2, and 08 in T3, 12 mm in T4. Klebsiella pneumonia showed the zone of inhibition as 18 mm in T1, T2 and T3 is 10 mm, 12 mm in T4 respectively. The positive control, ampicillin has showed the highest level of 40 mm when compared to sample taken.

According to the Table 3, variety T2 showed significant antibacterial and antifungal activities in all pathogens tested in different concentrations. The G+ve bacteria Staphylococcus aureus, G-ve bacteria Eschiershea coli was significantly inhibited in methanolic extract of all varieties with the standard in accordance with earlier report ³⁰ and its bioactive compounds can be used as chemo preventive agents to treat bacterial diseases without risk assessment ³¹.

TABLE 3: ANTIMICROBIAL ACTIVITY

CONCLUSION: In the current investigation, the methanol extracts of different varieties of Terminalia catappa leaves showed high antibacterial activity, exactly to standard whereas antifungal activity was half to the standard. Therefore the antibacterial activity of four different varieties is equally important and can be used for clinically isolated microorganisms as compared with the standard. The present study justified that the different varieties of Terminalia catappa leaves can be used in the traditional system to treat various infectious diseases caused by microbes especially by two microorganisms like staphylococcus aureus (a Gram -positive bacteria), Eschiershea coli (Gram -negative bacteria) were more susceptible and for the further investigations in the potential discovery of new natural bioactive compounds.

ACKNOWLEDGEMENT: The author of this paper is thankful to Surana College, South End, Bangalore, India for providing support.

CONFLICT OF INTEREST: Nil

REFERENCES:

1. Nair R and Chanda S: Antimicrobial activity of Terminalia catappa, Manilkara zapota and Piper betel Leaf extract. Indian Journal of Pharmaceutical Sciences 2008; 70(3): 390-393.
2. Edward F and Dennis GW: Tropical Island Terminalia catappa a series of the publication of the Environmental Horticulture Department Florida Fact sheet 1964; ST626.
3. Wright MH, Arnold MSJ, Lee CJ, Courtrey R, Anthony, Greese C and Cock IE: Evaluation of Indian Terminalia species against the pharyngitis causing Streptococcus pyogene. Pharmacognosy Communications 2016; 16(2): 85-92.
4. Oni O and Bada SO: Effects of seed size on seedling vigour in Terminalia ivorensis. Journal of Tropical Forest Science 1982; 4: 215-224.
5. Neelavathi P, Venkatalakshmi P and Brinda P: Anti-bacterial activities of aqueous and ethanol extracts of Terminalia catappa and bark against some pathogenic Bacteria. International Journal of Pharmacy and Pharma-ceutical Sciences 2013: 5(1).
6. Venkatalakshmi P, Brinda P and Vellangiri V: In-vitro antioxidant and anti-inflammatory studies on bark, wood, fruit of Terminalia catappa. International Journal of Phytomedicine, 2015; 7(3).
7. Godghate AG, Sawant RS and Jadhav SD: Phytochemical analysis of acetonic extract of fruits of Anacardium occidentals (Cashew nut) and Terminalia catappa (Almond). Asian Journal of Plant Science and Research 2013; 3(2): 150-153.
8. Yazdi AMM and Kamalinejad M, Antibacterial activity of Terminalia catappa against bacteria isolated from burn wounds and comparison with effects of selective antibiotics in-vitro. International Journal of Infectious Diseases 2008; 12(1): e410.
9. Kankia H: Phytochemical screening and antibacterial activities of leaf extracts of Terminalia catappa. Inter-national Journal of Science and Research 2012; 3: 358.
10. Wee YCA: Guide to medicinal plants. Singapore Science Centre, Description, Chemical Compounds and Uses 1992; 146.
11. Faruque MO, Uddin SB, Barlow JW, Hu S, Dong S, Cai Q, Li X and Hu X: Quantitative ethanobotany of medicinal plants used by indegenous communities in the Bandar ban district of Bangladesh, Frontiers in Pharmacology 2018; 9: 40.
12. Tan GT, Pezzuto JM, Kinghorn AD and Hughes SH: Evaluation of natural products as inhibitors of Human Immunodefieciency Virus type-1 (HIV-1) reverse transcriptase. Jou of Natural Products 1991; 54(14): 3-154.
13. Yonemoris MT, Oyama Y, Takeda Y, Tanaka T, Andoh T, Shinohara A and Nakata M: Evaluation of the anti-oxidant activity of environmental plants activity of the leaf extracts from seashore plants. Journal of Agricultural Food Chem 1999; 47(4): 1749-1754.
14. Krishnaveni M, Krishnakumari G, Banu CR and Kalaivani M: GC-MS Analysis of phytochemical in Terminalia catappa Antimicrobial assay. Indo American journal of pharmaceutical Research 2015; 5(3).
15. Krishnaveni M, Krishnakumari G, Banu CR and Kalaivani M: GC-MS Analysis of phytochemical in Terminalia catappa Linn. Antimicrobial assay, American Journal of Chinese Medicine 1997; 25 (2): 153-61.
16. Chandra S, Rakholiya K and Nair R: In-vitro interaction of certain antimicrobial agents in combination with plant extracts against some pathogenic bacterial strains. Asian Pacific Journal of Tropical Biomedicine 2012; 2(2): S876-S880.
17. Nagappa AN, Thakurdesai PA, Venkat RN and Singh J: Antidiabetic activity of Terminalia catappa Journal of Ethanopharmacology 2003; 88: 45-50.
18. Yazdi AMM and Kamalinejad M: Antibacterial activity of Terminalia catappa against bacteria isolated from burn wounds and comparison with effects of selective antibiotics in-vitro. International Journal of Infectious Diseases 2008; 12 (1): e410.
19. Ram J: Phytochemical screening and reported biological activities of some medicinal plants of Gujarat region. Jou of Pharm and Phyto 2015; 4(2): 192-198.
20. Noel ZG, Koffi NG, Justin KND, Kiyinima C and Joseph DA: Evaluation and Comparison of antifungal activities of Terminalia catappa and Terminalia mantaly (Com-bretaceae) on the in-vitro growth of Aspergillus fumigates. Journal of Medicinal Plants 2012; 6(12): 2299-2308.
21. Varkey IC and Kasturi MG: HPTLC analysis of stem Bark extracts of Terminalia chebula For alakoloid profile, Asian Journal of Biochemistry 2016; 11: 97-103.
22. Mandloi S, Srinivas R, Mishra R and Varma R: Antifungal activity of alcoholic leaf extracts of Terminalia catappa and Terminalia arjuna on some pathogenic and allergenic Fungi. Advances in Life Science and Technology 2013; 8: 25-27.
23. Ramachandramoorthy T, George MS and Balasubramani-yam S: Phytochemical screening and antibacterial activity of ethanolic extracts of Terminalia catappa International Journal of Research in Pharmacy and Chemistry 2016; 6(2): 345-349.
24. Rakholiya K, Chanda S: In-vitro interaction of certain antimicrobial agents in combination with plant extract against some bacterial strains. Asian pacific Journal of Tropical Biomedicine 2012; 2(2): S876-880.
25. Unnikrishna G, Shwetha V, Bhangale A and Patil S: Microbial sensitive assay of Terminalia catappa and Adathoda vasica. Journal of Biomedical and Biomedical Research 2014; 3(4): 58-62.
26. Sofowara A, Ogunbodede E and Onayade A, African Journal of Traditional Complementry Alternative Medicine 2013; 10(5): 210-229.
27. Evans WC: Trease and Evans Pharmacognosy, W.R. Sauders, London, Edition15^th, 2002; 137-140.
28. Slinkard K and Singleton VL: Total phenol analyses: automation and comparison with manual methods. Ame Journal of Enology and Viticulture 1977; 28(1): 49-55.
29. Charng-charng, Chyau, Pci and Mau TJ: Antioxidant properties of aqueous extracts from three different leaves of Terminalia catappa 2006; 39(10): 1099-1108.
30. Manzur A, Raju A and Rahman S: Terminalia catappa extracts against some pathogenic microbial strains, Pharmacology and Pharmacy 2011; 2: 299-305.
31. Babayi HI, Okujun KJI and Ijan UJJ: The antimicrobial activities of methanolic extracts of Eucalyptus camaldeulensis and Terminalia catappa against some pathogenic Microorganisms. Biochemistry 2004; 16(2): 106-111.
    

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    How to cite this article:

    Tasneem MIF and Narsegowda PN: Antimicrobial activity of different varieties of Terminalia catappa leaves. Int J Pharm Sci & Res 2018; 9(10): 4430-35. doi: 10.13040/IJPSR.0975-8232.9(10).4430-35.

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