An in vitro Evaluation of the Antimicrobial Activity of Curcuma longa against Selected Pathogenic Microorganisms

Nidhi Rao, Sandhya Mittal

Suresh Gyan Vihar University, Jaipur. India

*Corresponding Author E-mail: nidhiscorpian03@gmail.com

ABSTRACT:

The use of plants in healing of dermatophytes, burns along with infectious diseases is frequent in conventional medicine. The expansion of new antimicrobial agents against defiant pathogens is of escalating interest. Curcuma longa (turmeric) is a diminutive rhizomatous perennial herb of Zingiberaceae (Ginger family) originates from south eastern Asia, mainly from India. It has an elongated history of therapeutic uses as it is credited with a diversity of imperative beneficial properties. Therefore, the methanolic extract of Curcuma longa used locally in folk medicine was evaluated for antibacterial activity against different bacteria i.e. Shigella flexneri, Staphylococcus aureus, Salmonella typhi, Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus vulgaris, Enterobacter aerogenes, Aspergillus niger, Candida albicans, Trichophyton rubrum by disc diffusion method. When screening for antimicrobial activity, the results were expressed in terms of the diameter of the inhibition zone: The maximum efficacy of methanolic extract was showed against Shigella flexneri and Candida albicans.

KEYWORDS: Curcuma longa, Antibacterial activity, Antifungal activity, Plant extract.

INTRODUCTION:

Currently, multiple drug resistance has built-up due to the arbitrary use of commercial antimicrobial drugs frequently used in the treatment of infectious diseases^1,2. In accumulation to this predicament, antibiotics are occasionally coupled with adverse effects on the host together with immune suppression, hypersensitivity as well as allergic reactions³. There is an invariable necessitate for new moreover effectual therapeutic agents⁴. Consequently, there is a need to expand unconventional antimicrobial drugs for the healing of communicable diseases from medicinal plants^5,6.

Curcuma longa L., commonly known as turmeric, a deep yellow-to-orange powder that comes from the underground stems of the tropical perennial herb Curcuma longa of the family Zingiberaceae⁷. The powder of turmeric which is imitative from the rhizome of Curcuma longa, is generally used as a spice, food preservative, as well as food-colouring agent⁸. It also has an extensive history of restorative uses.

All over the world, it is one of large indispensable spices with a long in addition to distinguished human use predominantly in the Eastern civilization⁹. A few drops of turmeric juice obtained from the rhizomes can generate everlasting stain on the clothes. The source of the plant is not convinced, but it is contemplation to be originated from the south eastern Asia, most probably from India and further this plant is cultivated in all parts of India¹⁰. Turmeric is also cultivated in southern China, Taiwan, Burma, Japan, along with Indonesia¹¹ in addition to the way through the African continent¹². Turmeric powder has placid fragrance somewhat reminiscent of orange and ginger and also has a peppery bitter flavor and also best known as one of the major ingredients worn to make the curry spice.

The present study was done to evaluate the antimicrobial potential of methanolic extract of Curcuma longa. In this study, the drug has been used due to the several bacterial infections. They have been screened for their antibacterial activity against Shigella flexneri, Staphylococcus aureus, Klebsiella pneumonia, Proteus vulgaris, Enterobacter aerogenes, Chryseobacteriu gleum, Bacillus subtilis as well as the antifungal activity against Candida albicans, Aspergillus niger, Aspergillus fumigates, Aspergillus flavus. The antimicrobial activity of the plant extract was compared with standard antibacterial drug Tetracycline.

MATERIALS AND METHODS:

Collection:

Authentic samples: Various market samples of Curcuma longa were procured from Chunnilal Attar Ayurvedic Store, Ghat Gate, Jaipur in the month of March, 2010.

Identification:

All the samples were authenticated and were given identification number. The identification was as follows:

These samples were authenticated and submitted in Ethnomedicinal Herbarium, Centre of Excellence funded by DST, MGiaS, Jaipur (Rajasthan).

Processing of plant materials:

During the course of the study each sample was screened for its foreign matter and milled, before use.

Experimental details:

Present studies were performed on Curcuma longa for the antimicrobial studies.

ANTIMICROBIAL ACTIVITY:

Sources of test organisms:

Bacteria-Pure culture of all test organisms, namely Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Salmonella typhi, Shigella flexneri, Proteus vulgaris, Enterobactor aerogenes and fungi Candida albicans, Aspergillus niger, Trichophyton rubrum were obtained through the courtesy of Mahatma Gandhi Institute of Applied Sciences (MGiaS), Jaipur, which were maintained on Nutrient broth media. Culture of test microbes: For the cultivation of bacteria, Nutrient Agar Medium (NAM) was prepared by using 20 g Agar, 5 g Peptone, 3 g beef extract and 3 g NaCl in 1 L distilled water and sterilized at 15 lbs pressure and 121°C temperature for 25-30 min. Agar test plates were prepared pouring approximately 15 ml of NAM into the Petri dishes (10 mm) under aseptic conditions. A saline solution was prepared (by mixing 0.8% NaCl) in distilled water, followed by autoclaving and the bacterial\ cultures were maintained on this medium by regular sub-culturing and incubation at 37°C for 24-48 h. To prepare the test plates, in bacteria, 10-15 ml of the respective medium was poured into the Petri plates and used for screening. For assessing the bactericidal efficacy, a fresh suspension of the test bacteria was prepared in saline solution from a freshly grown Agar slant.

Preparation of test extracts:

Crushed powder (50 g) of all the species were successively soxhlet extracted with Methanol. Later, the homogenates was filtered and the residue was re-extracted twice for complete exhaustion, the extract was pooled. The filtrate was concentrated to dryness in vitro and re dissolved in respective solvents, out of which 80mg/10disc i.e. 8mg/disc concentration were stored at 4°C in a refrigerator, until screened for antibacterial activity.

Bactericidal assay:

For both, bactericidal in vitro Disc diffusion method was adopted (Gould and Bowie, 1952), because of reproducibility and precision. The different test organisms were proceeded separately using a sterile swab over previously sterilized culture medium plates and the zone of inhibition were measured around sterilized dried discs of Whatman No.1 paper(6 mm in diameter), which were containing 8 mg of the text extracts, its control (of the respective solvent) and tetracycline as reference drugs(standard disk) separately. Such treated discs were air-dried at room temperature to remove any residual solvent, which might interfere with the determination, sterilized and inoculated. These plates were initially placed at low temperature for 1 h so as to allow the maximum diffusion of the compounds from the test disc into the agar plate and later, incubated at 37°C for 24 h in case of bacteria, after which the zones of inhibition could be easily observed. Five replicates of each test extract were examined and the mean values were then referred.

The inhibition zone (IZ) in each case were recorded and the activity index (AI) was calculated as compared with those of their respective standard reference drugs (AI = Inhibition Zone of test sample / Inhibition zone of standard).

RESULTS AND DISCUSSION:

Antimicrobial Activity:

Antibacterial Activity of Curcuma longa (Turmeric):

When antibacterial activity of Curcuma longa was performed against above seven microorganisms through the preparation of alcoholic extract of plant and disc of .01mg/ml, 1mg/ml and 2mg/ml was prepared, the result of Curcuma longa antibacterial activity were appreciable but was maximum against Shigella flexneri (12 mm) and minimum against Klebsiella pneumoniae (6mm) and showed no activity against Proteus vulgaris and Staphylococcus aureus.

Fig 1: Graph showing Antibacterial Activity of Curcuma longa at different concentration.

Table 1: Showing Inhibition zone of Curcuma longa on different concentrations.

Microorganism	Inhibition zone(mm)
Microorganism	A(0.01mg/ml)	B(0.1mg/ml)	C(0.2mg/ml)
Proteus vulgaris	-	-	-
Staphylococcus aureus	-	-	-
Klebsiella pneumoniae	6	-	10
Shigella flexneri	6	12	11
Chryseobacteriu gleum	6	7	9
Enterobacter aerogenes	8	10	11
Bacillus subtilis	10	-	-

Antifungal Activity of Curcuma longa (Turmeric):

When antifungal activity of Curcuma longa was performed against above four fungus it was found that activity was quiet good but shown positive activity against Candida albicans (13mm) and no activity was shown against Aspergillus flavus, Aspergillus niger and Aspergillus fumigates.

Fig 2: Graph showing Antifungal Activity of Curcuma longa at different concentration.

A Fig (Against Klebsiella pneumonia)

B Fig (Against Chryseobacteriu gleum)

C Fig (Against Bacillus subtilis)

Figures showing Antibacterial activity of Curcuma longa (Turmeric) against select test organisms

Table 2: Showing Inhibition zone of Curcuma longa on different concentrations with standard.

Micro organism	I.Z of Standard	Inhibition zone(mm)
		A(.01mg/ml)	B(0.1mg/ml)	C(0.2mg/ml)
		IZ AI	IZ AI	IZ AI
*Proteus vulgaris*	19	-	-	-
*Staphylococcus aureus*	22	-	-	-
*Klebsiella* *pneumoniae*	14	6 0.42	-	10 0.71
*Shigella* *flexneri*	13	6 0.46	12 0.92	11 0.84
*Chryseobacteriu* *gleum*	18	6 0.33	7 0.38	9 0.50
*Enterobacter* *aerogenes*	16	8 0.50	10 0.62	11 0.68
*Bacillus subtilis*	21	10 0.47	-	-

I.Z=Inhibition zone of microorganisms A.I=Activity index A.I=I.Z of successive/I.Z 0f standard

Table 3: Showing Inhibition zone of Curcuma longa on different concentrations

Microorganism	Inhibition zone(mm)
Microorganism	A (0.01mg/ml)	B (0.1mg/ml)	C(0.2mg/ml)
*Candida albicans*	13	-	7
*Aspergillus* *niger*	-	-	-
*Aspergillus* *fumigates*	-	-	-
*Aspergillus* *flavus*	-	-	-

Table 4: Showing Inhibition zone of Curcuma longa on different concentrations with standard

Micro organism	I.Z of Standard	Inhibition zone(mm)
		A(.01mg/ml)	B(0.1mg/ml)	C(0.2mg/ml)
		IZ AI	IZ AI	IZ AI
*Candida albicans*	12	13 1.08	- -	7 0.58
*Aspergillus* *niger*	16	- -	- -	- -
*Aspergillus* *fumigates*	19	- -	- -	- -
*Aspergillus* *flavus*	9	- -	- -	- -

CONCLUSION:

From the above studies it can be accomplished that the methanolic extract of Curcuma longa possesses the competent antimicrobial substances. It may be diminish on the basis of using this plant’s extract as folkloric remedies. The results of antibacterial activity were quite good i.e. Shigella flexneri showed very good results and minimum against Klebsiella pneumonia whereas antifungal activity of Curcuma longa was found that activity was appreciable but showed positive activity against Candida albicans. It shows that this plant have potentials role in future as drug or therapeutic targets.

ACKNOWLEDGEMENT:

Authors would take pride in rendering our deep sense of gratitude to beloved Chairperson Shri Sunil Sharma, Chief Mentor Dr. Shudhanshu, Vice Chancellor Dr. D. N. Rao for their constant encouragement and ever willingness to provide the necessary facilities at the Institution. Authors wish to express the sincere thanks to all the office staff for their constant support and encouragement throughout this work.

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Received on 31.12.2013 Modified on 20.04.2014

Research J. Science and Tech. 6(2): April- June 2014; Page 71-74