Indian medicinal herb: Antimicrobial efficacy of Mesua ferrea L. seed extracted in different solvents against infection causing pathogenic strains

Sumitra Chanda, Kalpna Rakholiya, Jigna Parekh

Phytochemical, Pharmacological and Microbiological Laboratory, Department of Biosciences (UGC-CAS), Saurashtra University, Rajkot 360 005, Gujarat, India

Indian medicinal herb: Antimicrobial efficacy of Mesua ferrea L. seed extracted in different solvents against infection causing pathogenic strains

Sumitra Chanda*, Kalpna Rakholiya, Jigna Parekh

Phytochemical, Pharmacological and Microbiological Laboratory, Department of Biosciences (UGC-CAS), Saurashtra University, Rajkot 360 005, Gujarat, India

Antimicrobial activity

Mesua ferrea L.

Seed

Organic solvents

Microorganism

Objective: To study the antimicrobial potential of Mesua ferrea (M. ferrea) L. seed extracts employed for antimicrobial assay. Methods: The plant powder was extracted in seven different solvents of increasing polarities against a wide spectrum of microbial strains. Agar disc diffusion method was employed for antimicrobial assay at the concentration of 500 μg/disc. Gram-positive bacteria were most susceptible and yeast was most resistant. The pronounced antimicrobial activity was with the extracts in non-polar solvents than in polar ones. Results: The results were compared with the zones of inhibition produced by commercially available standard antibiotics. The lipophilic extracts of M. ferrea L. showed more activity towards Gram positive bacteria. Conclusion: These results indicate that activity could be attributed to the presence of essential oil, xanthones and coumarines present within the seed of this plant. These results indicate that M. ferrea extract could be useful in fighting emerging drug-resistant microorganisms.

1. Introduction

Herbal medicine represents one of the most important fields of traditional medicine all over the world. The widespread use of herbal remedies and healthcare preparations, as those described in ancient texts such as the Vedas and the Bible, and obtained from commonly used traditional herbs and medicinal plants, has been traced to the occurrence of natural products with medicinal properties. To promote the proper use of herbal medicine and to determine their potential as sources for new drugs, it is essential to study medicinal plants, which have folklore reputation in a more intensified way[1-6].

Recently, infections caused by microorganisms have increased tremendously and antibiotic resistance has become a global therapeutic problem. On account of this, special attention has been paid to extracts and biologically active compounds isolated from plant species[7,8] with a hope to get new promising drugs to treat these resistant stubborn microorganisms.Antimicrobials of plant origin are not only efficient in the treatment of infectious diseases but they are free from many of the side effects that are often associated with synthetic antimicrobials[9]. Natural products of higher plants may give a new source of antimicrobial agents with possibly novel mechanisms of action[10,11].

Mesua ferrea (M. ferrea) L. commonly known as Nagkesar belongs to the family Guttiferae. It is a medium sized glabrous tree found in the North-East and Southern part of India. The trunk is straight, erect and ash colored. The bark is grayish or reddish-brown. The leaves are oblong-lanceolate or acute. Flowers are large, white and fragrant. The fruits are ovoid and the seeds are angular, smooth and chest-nut brown[12]. The principal constituents of M. ferrea include mesuaferrone-A & B, mesuaferrol, mesuanic acid, α-&β-amyrin and β-sitosterol present in the stamen[13] while it is reported that seeds contain essential oils, xanthones and coumarins[14-16]. Traditionally this plant is widely used for curing many ailments in India. Decoction of 2-3 flowers with sugarcandy is given twice a day for 3 d to stop bloody stool. Powder of flowers and fruits with butter is applied locally on piles. Essential oil is used to treat skin diseases and its application is recommended in rheumatism[17]. The few scientific reports on thebiological activities of M. ferrea studied in the literature include antiasthmatic activity[18], anti-inflammatory and C.N.S. dependent activities[19], estrogenic and progestational activity[20], antispasmodic activity[21], antimicrobial and anthelmintic activity of essential oil[22], as bactericide from leaf[23] and antimicrobial activity of flower[20-25]. M. ferrea is also used in herbal formulation to treat piles[26] and cancer[27]. In-vivo antioxidant and immunomodulatory activity[28] as well as disinfection studies of seed kernel oil are reported[29]. According to Mathekga and Mayer[30], in vitro antimicrobial screening methods are useful tools to select active extracts for further chemical and pharmacological investigations.

This paper reports the first attempt to study the antimicrobial property of Indian medicinal plant, M. ferrea L. seed (Guttiferae) extracts in different solvents against an array of human pathogens.

2. Material and methods

2.1. Plant material

M. ferrea L. seeds were purchased from local market of Rajkot, India. The taxonomic identity of this plant was confirmed by Dr. N. K. Thakrar Department of Biosciences, Saurashtra University, Rajkot. The material was purchased in the form of dried part, in July, 2010 which was then homogenized to fine powder and stored in airtight bottles.

2.2. Extraction of plant material

The air-dried and powdered plant material (10 g of each) was extracted with 100 mL each of petroleum ether, 1, 4-dioxan, chloroform (CHCl3), acetone, N, N, dimethyl formamide, ethanol (CH3OH) and water (H2O), kept on a rotary shaker for 24 h. Thereafter it was filtered and centrifuged at 5 000 g for 15 min. The supernatant was collected and evaporated to dryness to give the crude dried extract. The extractive yield (%) of all the extracts is shown in Table 1.

2.3. Microbial strains

The test microbial strains investigated are listed in Table 1. All the bacterial strains were obtained from National Chemical Laboratory (NCL), Pune, India. The bacterial strains were grown in the nutrient broth and maintained on nutrient agar slants at 4 ℃while fungal strains were grown in Sabouraud broth and maintained on MGYP slants (yeast) and potato dextrose agar slants (mould) at 4℃

2.4. Antimicrobial assay

A modified agar disc diffusion method[31,32] was used to determine the antimicrobial activity. Molten Mueller Hinton agar No. 2 (HiMedia) was inoculated with microbial cell suspension (100 μL) and poured into sterile Petri dishes. Sterile filter paper discs of 7 mm diameter were impregnated with 20 μL extract solution equivalent to 500 μg of the each dried extract in 100% DMSO (Dimethylsulphoxide) and air dried. Thereafter the discs were placed on the surface of the seeded agar plates. Piperacillin (100 μg/disc), Gentamicin (10 μg/ disc) and Amphotericin B (100 units/disc) were used as positive controls. Paper discs loaded with 20 μL of DMSO served as negative control. The plates were incubated at 37 ℃ for 24 h for all the bacterial strains while that of fungal strains were incubated at 28 ℃ for 48 h. The experiment was done three times to minimize error. After incubation period the antibacterial activity was evaluated by measuring the inhibition zones. An inhibition zone of 14 mm or greater (including diameter of the disc) was considered as high antibacterial activity.

3. Results

The presence of antifungal and antibacterial substances in the higher plants is well established. Plants have provided a number of novel drugs and they have made significant contribution towards health. The extractive yield of different solvents is given in Table 1. Maximum yield was obtained in DMF (23.18%) while minimum was in petroleum ether (3.42%) (Table 1).

In the present study an array of solvents with different polarity were used to study the antibacterial activity of M. ferrea seed extract. The antibacterial activity was definitely better when the plant was extracted in organic solvents than in aqueous solution (Table 1). Out of the 9 Gram positive strains studied, significant antibacterial activity was shown against M. flavus followed by B. subtilis and S. subflava. Lowest antibacterial activity was shown against S. epidermidis. M. ferrea leaf extract in all the organic solvents and aqueous showed activity against the Gram positive bacteria studied. An entirely different trend was obtained with Gram negative bacteria. Here only 35% of the strains were active and maximum antibacterial activity was shown against P. putida followed by P. mirabilis and K. aerogenes; all the other strains viz. A. fecalis, C. freundii, E. aerogenes, E. coli, P. vulgaris, P. aeruginosa, P. pseudoalcaligenes, P. testosteroni and S. typhimurium were resistant.

In antifungal study, except T. beigelli, none of the yeast showed antifungal activity; on the other hand, 50% of the moulds studied showed antifungal activity. Only, chloroform, DMF and ethanol extracts showed activityagainst A. candidus and M. hiemalis wehmer, while the other three organic solvent extracts (petroleum ether, 1, 4-dioxan and ethanol) did not show any antifungal activity. Aqueous extract showed slight activity against A. candidus while it was ineffective against all the other studied moulds. There is no antifungal activity found due to the fungal cell wall is a complex structure and extensive cross-linking between chitin, glucans and other polymers.

Table 1Antimicrobial efficacy of M. ferrea L. seed extracts against microbial strains.

The results of the antibacterial and antifungal activity of the different extracts of M. ferrea were compared with the standard antimicrobics (Table 1). The extracts showed significant activity against more than 50% of the investigated microbial strains, which is infect a promising result, which was also comparable with standard antimicrobics, which suggest that the plant extracts contain certain constituents with antimicrobial properties that can be used as antimicrobial agents in new drugs for the therapy of infectious diseases caused by pathogens. It is interesting to note that the extracts are not pure compounds and in spite of it good results were obtained which only suggests the potency of these extracts.

4. Discussion

From the results it is observed that the antibacterial activity was more pronounced with the extracts in nonpolar solvents than in polar ones. This suggests that the polarity of the solvent plays an important role in exhibiting antibacterial activity[33]. The Gram negative bacteria were more resistant than Gram positive bacteria as also reported by many researchers[34-36]. Also the organic solvents extracts of M. ferrea seed were found to have good activity against bacteria than yeast or moulds. This differences in the activity may be attributed to the fact that the cell wall in Gram positive bacteria is of a single layer, whereas the Gram negative cell wall is multilayered structure and the yeast cell wall is quite complex[37,38]. These results are in good agreement with the results reported earlier by Ali et al[39].

Antifungal activity is not common in medicinal plants. Marting et al[40] screened 23 extracts of 12 Cuban plants but they did not inhibit the growth of yeast. The potential for developing antimicrobial from higher plants is rewarding as it will lead to the development of a phyto-medicine to act against microbes. Plant based antimicrobials have enormous therapeutic potential as they can serve the purpose without any side effects as compared to synthetic compounds. The results obtained are encouraging as the organic solvent extracts have shown considerable antibacterial activity[41,42].

The present work has demonstrated the antimicrobial potential of M. ferrea L. seed extracted in various solvents. This is the first report on the antimicrobial activity of M. ferrea. The above observations have clearly demonstrated M. ferrea, as a folk remedy which confirms its folkloric utilization. This study provides scientific insight to further determine the antimicrobial principles and investigate other pharmacological properties. Hence, M. ferrea seed extract should be taken up for further bioactivity guided isolation of active compound and carry out pharmaceutical studies.

Conflict of interest statement

We declare that we have no conflict of interest.

Acknowledgement

The authors thank Prof. S.P. Singh, Head, Department of Biosciences, Saurashtra University for providing excellent research facilities; and University Grants Commission, New Delhi, India (F. No. 37-524/2009 (SR)) for providing financial support in the form of a Major Research Project.

[1] Chanda S, Kaneria M. Indian nutraceutical plant leaves as a potential source of natural antimicrobial agents. In: Mendez-Vilas A, editor. Science against microbial pathogens: communicating current research and technological advances. Spain: Formatex Research Center; 2011.

[2] Hemalatha M, Thirumalai T, Saranya R, Elumalai EK, David E. A review on antimicrobial efficacy of some traditional medicinal plants in Tamilnadu. J Acute Dis 2013; 99-105.

[3] Fernandez-Agullo A, Pereira E, Freire MS, Valentao P, Andrade PB, Gonzalez-Alvarez J, et al. Influence of solvent on the antioxidant and antimicrobial properties of walnut (Juglans regia L.) green husk extracts. Ind Crop Prod 2013; 42: 126-132.

[4] Baravalia Y, Kaneria M, Vaghasiya Y, Parekh J, Chanda S. Antioxidant and antibacterial activity of Diospyros ebenum Roxb. leaf extracts. Turk J Biol 2009; 33: 159-164.

[5] Sanchez-Burgos JA, Ramirez-Mares MV, Larrosa MM, Gallegos-Infante JA, Gonzalez-Laredo RF, Medina-Torres L, et al. Antioxidant, antimicrobial, antitopoisomerase and gastroprotective effect of herbal infusions from four Quercus species. Ind Crop Prod 2013; 42: 57-62.

[6] Chanda S, Rakholiya K, Dholakia K, Baravalia Y. Antimicrobial, antioxidant, and synergistic properties of two nutraceutical plants: Terminalia catappa L. and Colocasia esculenta L. Turk J Biol 2013; 37: 81-91.

[7] Chanda S, Rakholiya K, Nair R. Antimicrobial activity of Terminalia catappa L. leaf extracts against some clinically important pathogenic microbial strains. Chinese Med 2011; 2: 171-177.

[8] Jeyaseelan EC, Jenothiny S, Pathmanathan MK, Jeyadevan JP. Antibacterial activity of sequentially extracted organic solvent extracts of fruits, flowers and leaves of Lawsonia inermis L. from Jaffna. Asian Pac J Trop Biomed 2012; 2: 798-802.

[9] Chanda S, Rakholiya K. Indian Combination therapy: Synergism between natural plant extracts and antibiotics against infectious diseases. In: Mendez-Vilas A, editor. Science against microbial pathogens: Communicating current research and technological advances. Spain: Formatex Research Center; 2011.

[10] Kaneria M, Chanda S. Evaluation of antioxidant and antimicrobial capacity of Syzygium cumini L. leaves extracted sequentially in different solvents. J Food Biochem 2013; 37: 168-176.

[11] Kaneria M, Baravalia Y, Vaghasiya Y, Chanda S. Determination of antibacterial and antioxidant potential of some medicinal plants from Saurashtra region, India. Indian J Pharm Sci 2009; 71: 406-412.

[12] Anandakumar A, Balasubramaniam M, Muralidharan R. Nagakesara - a comparative pharmacognosy. Ancient Sci Life 1986; 5: 263-268.

[13] Subramanyam Raju M, (late) Subba Rao NV. Chemical study of the stamens of Mesua ferrea Linn. J Recher Imagerie Med 1977; 12: 124-126.

[14] Subramanyam Raju M, (late) Subba Rao NV. Fatty acid composition of nahor (Mesua ferrea Linn) seed oil. J Recher Imagerie Med 1977; 12: 97-99.

[15] Banerjee R, Choudhary AR. Mesua ferrea: chemical constituents and biological activity. J Chem Soc Pakistan 1993; 15: 207-211.

[16] Dennis TJ, Akshaya Kumar K. Constituents of Mesua ferrea -A review. Fitoterapia 1998; 69: 291-304.

[17] Manandhar NP. Medicobotany of Borkha District, Nepal - An elucidation of medicinal plants. Int J Crude Drug Res 1990; 28: 17-25.

[18] Bhide MB, Naik PY, Joshi RS. Studies on the antiasthmatic activity of Mesua ferrea. Bull Haffkine Inst 1977; 5: 27-30.

[19] Gopalakrishnan C, Shankaranarayana D, Nazimudeen SK, Viswanathan S, Kameswaran L. Anti-inflammatory and C.N.S. dependent activities of xanthones from Calophyllum inophyllum and Mesua ferrea. Indian J Pharmacol 1980; 12: 181-191.

[20] Meherji PK, Chetye TA, Munshi SR, Vaidya RA, Antarkar DS, Koppikar S, et al. Screening of Mesua ferrea Linn. (nagkeshar) for estrogenic and progestational activity in human and experimental models. Indian J Exp Biol 1978; 16: 932-933.

[21] Prasad DN, Basu SP, Srivastava AK. Antispasmodic activity of the crude and purified oil of Mesua ferrea seed. Ancient Sci Life 1999; 19: 74-75.

[22] Kakrani HK, Nair GV, Dennis TJ, Jagdale MH. Antimicrobial and anthelmintic activity of essential oil of Mesua ferrea Linn. Indian Drugs 1984; 21: 261-262.

[23] Mazumder R, Dastidar SG, Basu SP, Kumar S. Emergence of Mesua ferrea Linn. leaf extract as a potent bactericide. Ancient Sci Life 2003; 22: 160-165.

[24] Mazumder R, Dastidar SG, Basu SP, Mazumder A, Singh SK. Antibacterial potentiality of Mesua ferrea Linn. flowers. Phytother Res 2004; 18: 824-826.

[25] Verotta L, Lovaglio E, Vidari G, Finzi PV, Neri MG, Raimondi A, et al. 4-alkyl & 4-phenylcoumarins from Mesua ferrea as promising multidrug resistant antimicrobials. Phytochem 2004; 65: 2867-2879.

[26] Sannd R, Bansal P, Bajwa RMS, Acharya MV. Folk medicine of Patiala for Arsha (piles). J Ayurveda Health 2004; 56: 525-529.

[27] Palani V, Senthilkumaran RK, Govindasamy S. Biochemical evaluation of antitumor effect of muthu marunthu (a herbal formulation) on experimental fibro sarcoma in rats. J Ethnopharmacol 1999; 65: 257-265.

[28] Chahar MK, Sanjaya Kumar DS, Lokesh T, Manohara KP. Invivo antioxidant and immunomodulatory activity of mesuol isolated from Mesua ferrea L. seed oil. Int Immunopharmacol 2012; 13: 386-391.

[29] Adewale AI, Mirghani MES, Muyibi SA, Daoud JI, Abimbola MM. Disinfection studies of Nahar (Mesua ferrea) seed kernel oil using pour plate method. Afr J Biotechnol 2011; 10: 18749-18754.

[30] Mathekga ADM, Meyer JJM. Antibacterial activity of South African Helichrysum species. South Afr J Bot 1998; 64: 239-295.

[31] Bauer AW, Kirby WMM, Sheriss JC, Turck M. Antibiotic susceptibility testing by standardized single method. Am J Clin Pathol 1966; 45: 493-496.

[32] NCCLS (National Committee for Clinical Laboratory Standards). Performance 545 standards for antimicrobial disk susceptibility test. 6th ed. Approved standard. M2-A6, 546 Wayne Pa, 1997.

[33] Parekh J, Karathia N, Chanda S. Evaluation of antibacterial activity and phytochemical analysis of Bauhinia variegate L. bark. Afr J Biomed Res 2006; 9: 53-56.

[34] Kiran SR, Sita P, Janardhan K. Evaluation of in vitro antimicrobial activity leaf and stem essential oils of Chloroxylon swietenia Dc. World J Microbiol Biotechnol 2008; 24: 1909-1914.

[35] Chanda S, Dudhatra S, Kaneria M. Antioxidative and antibacterial effects of seeds and fruit rind of nutraceutical plants belonging to the family fabaceae family. Food Funct 2010; 1: 308-315.

[36] Chanda S, Kaneria M, Vaghasiya Y. Evaluation of antimicrobial potential of some Indian medicinal plants against some pathogenic microbes. Indian J Nat Prod Res 2011; 2: 225-228.

[37] Chanda S, Baravalia Y, Kaneria M, Rakholiya K. Fruit and vegetable peels – strong natural source of antimicrobics. In: Mendez – Vilas A, editor. Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology. Spain: Formatex; 2010.

[38] Rakholiya K, Sumitra Chanda. In vitro interaction of certain antimicrobial agents in combination with plant extracts against some pathogenic bacterial strains. Asian Pac J Trop Biomed 2012; 2: S876-S880.

[39] Ali MA, Sayeed MA, Bhuiyan MSA, Sohel FI, Yeasmin MS. Antimicrobial screening of Cassia fistula and Mesua ferrea. J Med Sci 2004; 4: 24-29.

[40] Marting MJ, Betancourt J, Alonso-Gonzalez N, Jauregui A. Screening of some Cuban medicinal plants for antimicrobial activity. J Ethnopharmacol 1996; 52: 171-174.

[41] Mohanasundari C, Natarajan D, Srinivasan K, Umamaheswari S, Ramachandran A. Antibacterial properties of Passiflora foetida L. - a common exotic medicinal plant. Afr J Botechnol 2007; 6: 2650-2653.

[42] Chanda S, Kaneria M, Baravalia Y. Antioxidant and antimicrobial properties of various polar solvent extracts of stem and leaves of four Cassia species. Afr J Biotechnol 2012; 11: 2490-2503.

ment heading

10.1016/S2221-6189(13)60143-2

8 June 2013

*Corresponding author: Dr. SV Chanda, Phytochemical, Pharmacological and Microbiological Laboratory, Department of Biosciences (UGC-CAS), Saurashtra University, Rajkot-360 005, Gujarat, India.

E-mail: svchanda@gmail.com

ARTICLE INFO

Article history:

Received in revised form 15 July 2013

Accepted 28 July 2013

Available online 20 December 2013