MORINDA PUBESCENS BARK EXTRACTS SHOWS ANTIMICROBIAL ACTIVITY

MORINDA PUBESCENS BARK EXTRACTS SHOWS ANTIMICROBIAL ACTIVITY

Bharat Kumar Tyagi *, Rajesh Kumar Goyal and Pradeep Chauhan

Department of Pharmaceutical Chemistry, Institute of Professional Studies, College of Pharmacy, Shivpuri link road, Gwalior-474001, Madhya Pradesh, India.

ABSTRACT: Morinda Pubescens belonging to the family rubiaceae has been reported to posses many medicinal properties according to indigenous system of medicine. The search for biologically active compounds from natural sources has always been of great interest to researchers looking for new drugs useful in infectious disease. In the present study, the various extracts of Morinda Pubescens bark was tested for their antibacterial and antifungal activities against gram positive bacteria such as Staphylococcus aureus, Bacillus cereus, gram negative organisms such as Escherichia coli, Pseudomonas aeruginosa and fungi such as Candida albicans, Aspergillus niger respectively by cup plate method. All the extracts were effective against the tested microorganism which was comparable with standard antibiotics. The ethanolic extract showed highest zone of inhibition then other extracts tested. The zone of inhibition of the extracts of Morinda Pubescens indicating that the plant of Morinda Pubescens can fight these microorganism effectively due to the presence of phenol,triterpinoids and tannins it could be better alternative to the modern medicine

Morinda Pubescens, Bark extracts, antibacterial, antifungal

INTRODUCTION: India has a rich cultural heritage of traditional medicines which chiefly comprised the two widely flourishing system of treatment i.e Ayurvedic and Unani systems since ancient times. The multiple therapeutic action and uses of these drugs are sufficient described in classical literature on indigenous medicines in many medicinal plant books and pharmacopoeias ¹. For a long period of time, plants have been a valuable source of natural products for maintaining human health, especially in the last decade, with more intensive studies for natural therapies. Now a day, the use of phytochemical for pharmaceutical purpose has gradually increased in many countries

According to World Health Organization (WHO) medicinal plants would be the best source to obtain a variety of drugs. About 80% of individuals from developed countries use traditional medicine, which has compounds derived from medicinal plants ².

The antimicrobial compounds produced by plants are active against plants and human pathogenic microorganism ³. Medicinal plants represent a rich source of antimicrobial agents. Plants are used medicinally in different countries and are a source of many potent and powerful drugs ⁴. Infectious diseases caused by number of microorganisms are the world’s major threat to human health and account for almost 50,000 deaths every day ⁵.

 Morinda Pubescens (Rubiaceae) is an important medicinal plant of tropical and subtropical traditional uses.  It also occurs in Jhabua, Kerala, Karnataka, Pune, Maharashtra, Chennai, Tamil Nadu, Thanjavur District, drought prone area of Osmanabad, Andman and Nicobar Island. It is commonly known as ‘Indian Mulberry’ and aal in Hindi. Its medicinal usage has been reported in the Indian Traditional systems of medicine.

The various parts of this tree has been extensively used as- eczema, fever due to primary complex, ulcer,  glandular swellings, digestive disorders especially in children, in venereal diseases ⁶, rheumatic disease, dysentery ⁷, Antioxidant, cytotoxic activities, anticancer activity ⁸, antiplasmodial activity ⁹, nutritive analysis ¹⁰, antimicrobial activity and phytochemical screening ¹¹, antiplasmodial and Hepatoprotective ¹².

Literature survey revealed that the plant bark extract has yet not been screened for its traditional claim of antimicrobial activity. Therefore the objective of this work was to explore the antimicrobial properties of Morinda Pubescens bark.

MATERIAL AND METHOD:

Plant Material

The barks of Morinda Pubescens were collected from the area around the Jhabua district of Madhya Pradesh, India. The Botanical identity was confirmed by department of Botany, Jiwaji University, Gwalior, Madhya Pradesh, India.

Preparation of extract

The collected barks were shade dried at room temperature. The dried barks were size reduced to coarse powder (75gm) and macerated with petroleum ether, Chloroform, ethanol and distilled water separately for five days. The extracts of M. Pubescens barks were collected separately, filtered and concentrated under vacuum using rotary vacuum evaporator (Dolphin Instrument). The consistency, color, odour, taste and total yield (%) of the extracts of M. Pubescens bark was noted in Table 1. All the extracts were kept in desiccator(Dolphin Instrument) until further use.

TABLE 1: CONSISTENCY AND COLOR OF THE VARIOUS EXTRACTS OF MORINDA PUBESCENS BARK

Preliminary Phytochemical Analysis

Chemical tests below were carried out on the solid, semi-solid and sticky extracts of M. pubescens to determine the active constituents according to the procedures and methods in Trease and Evans (2009), Harborn (1998) and Kokate (2007). These tests were employed in detect the presence of secondary metabolites, such as alkaloids, glycosides, fixed oil & fats, flavonoids, phenol, triterpinoids and tannins in the plant under investigation in Table 2. ^{13, 14, 15}.

TABLE 2: PRELIMINARY PHYTOCHEMICAL STUDIES OF MORINDA PUBESCENS BARK EXTRACTS

Collection of microorganisms

Gram positive organism such as gram positive bacteria such as Staphylococcus aureus (MTCC 3160), Bacillus cereus (MTCC 9786), gram negative organisms such as Escherichia coli (MTCC 118), Pseudomonas aeroginosa (MTCC 4673) and fungi such as Candida albicans (MTCC 1637), Aspergillus niger (MTCC 282) were used for this study. All the bacterial cultures were obtained from Microbial Type Culture Collection & Gene Bank (MTCC), Sector 39-A, Chandigarh.

Antimicrobial activity

The antibacterial activity was studied by cup plate method as described by Indian Pharmacopeia 1996 and Anandarajagopal et al ^{16, 17}. All the extracts were dissolved separately in 2% v/v Tween 80 at a concentration of 10mg/ml. The respective bacterial culture was spread into the nutrient agar plates for uniform distribution of colonies. Using a sterile cork borer, 6mm wide well was made on each agar plates. All the extracts (10mg/ml) were poured separately into each well using a sterile micropipette and Ofloxacin (10µg/ml) was used as standard. The plates were incubated for 24 h at 37±2ºC. After incubation, the zone of inhibition was measured by Zone reader (Jyoti Scientific) and the values were tabulated in Table 3. All the experiments were done in triplicate.

TABLE 3: ANTIBACTERIAL ACTIVITY ZONE OF INHIBITION (mm)

Antifungal activity                                           

The antifungal activity was studied by cup plate method as described by Indian Pharmacopeia 1996 and Ramesh C. Ket al ^{16, 18}. First of all Sabouraud Dextrose Agar media was prepared, sterilized than spread into sterilized pertiplates. After solidification the Fresh cultures of Candida albicans and Aspergillus niger were introduced into Petri plates and BOD incubation at 25±2ºC for 96 hours.

Using a sterile cork borer, 6mm wide well was made on each agar plates. All the extracts were dissolved separately in 2% v/v Tween 80 at a concentration of 10mg/ml were poured separately into each well using a sterile micropipette and fluconazole (10µg/ml) was used as standard. The zone of inhibition was measured by Zone reader (Jyoti Scientific) and the values were tabulated in Table 4. All the experiments were done in triplicate.

TABLE 4: ANTIFUNGAL ACTIVITY ZONE OF INHIBITION (mm)

RESULT AND DISCUSSION:

The consistency, color, odour, taste and total yield of petroleum ether, chloroform, ethanol and aqueous extracts of M. pubescens were recorded in Table 1. Many medicinal plant extracts have been known to possess antimicrobial activity and are used for the treatment of microbial infections due to the presence of certain chemical constituents.

In the present study, the preliminary phytochemical investigation on M. pubescens extracts revealed the presence of various chemical constituents in the extracts was reported in Table 2.   Antimicrobial activity was carried against four different extracts from (bark part) plant M. pubescens (Rubiaceae), were tested against both gram positive and gram negative bacteria and fungus.

The results of the plant extracts viz., petroleum ether, chloroform, ethanol and Aqueous had significant antibacterial and antifungal activities among which ethanol extract (10mg/ml) exhibited maximum (14-17mm) zone of inhibition against all the tested microorganism whereas chloroform extract (10mg/ml) exhibited lower (09-13mm) zone of inhibition followed by petroleum ether extract (10mg/ml) which was well comparable with reference standards ofloxacin and fluconazole at the concentration of 10µg/ml respectively.

This study has shown the scientific basis for the therapeutic uses of traditional plant and confirmed its ethanolicmedicinal claims. The results mentioned revealed that M. pubescens possess considerable antimicrobial activity against selected microbial strains. The extracts of M. pubescens may be useful as an alternative antimicrobial agent as natural medicine for the treatment of diseases caused by microbes.

 CONCLUSION: The plant of M. pubescens (bark part) belonging to the family Rubiaceae was taken up for the study of activity against both gram positive and gram negative bacteria and fungi. The properties of these extracts encourage development a novel broad spectrum antimicrobial formation in future. The isolation of bioactive components of these extracts responsible for the activity is in progress. The extracts of M. pubescens may be useful as an alternative antimicrobial agent as natural medicine for the treatment of diseases caused by microbes.

ACKNOWLEDGMENT: Authors are thankful to Dr. Arun Kumar Tyagi, Director, IPS group of colleges, Gwalior (M.P.), India, for his constant support in conducting the study.

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