Anti-microbial Activity of Rhizome extracts of Kaempferia scaposa (Nimmo) Benth

Anti-microbial Activity of Rhizome extracts of Kaempferia scaposa (Nimmo) Benth
Dr. Sanjay Jagtap
Department of Botany, Elphinstone College, Mumbai.
ABSTRACT
Kaempferia scaposa is a perennial herb belonging to family Zingiberaceae, an important medicinal plant widely used in several indigenous medicinal formulations. Rhizome is a perennial organ with a number of primary and secondary metabolites. In present investigation the antimicrobial study of rhizome extracts of Kaempferia scaposa were evaluated against Salmonella typhi, Staphylococcus aureus, Escherichia coli and Aspergillus terreus. The methanolic extract result showed 14mm diameter zone of inhibition in Staphylococcus aureus, 12mm in Salmonella typhi. Similarly in acetone extract 2mm in Staphylococcus aureus, 6mm in Salmonella typhi and 1mm in Escherichia coli. Chloroform extract showed moderate zone of inhibition in Staphylococcus aureus and Escherichia coli. The present findings for antimicrobial activity suggested that the presence of bioactive compounds within rhizome were responsible for activity.
Key words: - Antimicrobial activity, Kaempferia scaposa, Secondary metabolites, Medicinal
plant.
Introduction
Kaempferia scaposa is an erect perennial herb 0.5 to 1.5 meter tall with fleshy sweet scanted rhizome belonging to family Zingiberaceae, an important medicinal plant widely used in several indigenous medicinal formulations. Distributed widely in peninsular and extra peninsular parts of India; generally found on floors of moist deciduous forests from 600 -750 m ; has been reported from Anamalai hills- Tamilnadu ; Hassan and Canada districts – Karnataka ; Ranchi – Orisssa ; Palmaujungke – Bhihar ; Kashia hills – Assam ; Dehradun – Haryana ; Punjab and Bengal. Lonawala in Pune district of Maharashtra (Hooker, 1882; Cooke, 1903; Duthie, 1911; Bamber, 1916; Fischer, 1921; Babu, 1977; Mathew, 1981 – 1983; Ugemuge, 1986; Yadav and Sardesai, 2010).
In Ayurveda, the important formulations using this herb are Cyavanaprāśam, Aśokāriṣṭam, Baladhātryādi tailam, Kalyanakaghritham, etc (Shivranjan, et al., 1994).The drug "Hallakam" prepared from this is in popular use in the form of powder or as an ointment application to wounds and bruises to reduce swellings. The tubers are useful in vitiated conditions of vāta and kapha, gastropathy, dropsy, inflammations, wound, ulcers, blood clots, tumours and cancerous swellings (Warrier, et al., 1995). Recently, Arambewela et al., (1999) reported that the essential oils of Kaempferia galanga root and rhizome showed antibacterial activity against Escherichia coli and Staphylococcus aureus.
Hence, several medicinal plants need to be necessary possible antimicrobial activity evaluation and to get admirable remedy for a microbial origin from a variety of ailments (Ubramani, et al.,2003).
In present investigation rhizome extracts were tested against Salmonella typhi, Staphylococcus aureus , Escherichia coli and Aspergillus terreus.

A :- Habit of Kaempferia scaposa, B:- Rhizome of Kaempferia scaposa.
Material and Methods
Sampling:
Fresh samples of rhizome of Kaempferia scaposa were collected from Lonavala, Dist., Pune region of Western Ghats of Maharashtra (Fig. 1 and 2). These plants were identified and authenticated using herbarium collection at Dept. of Botany, DST-FIST School of Life Science, SRTM University, Nanded (MS), India, Department of Botany Dr.Babasaheb Ambedkar Marathwada University, Aurangabad .Fresh rhizome were washed thoroughly under running tap water followed by sterile distilled water and dried under shade. The shade dried material was ground into coarse powder using mechanical grinder. This coarse powder was sieved by 1 mm pore size sieve. The powder was stored in airtight containers at room temperature till further antimicrobial activity.



Soxhlate extraction:
Exhaustive Soxhlate extraction was performed using a classical Soxhlate apparatus with
accurately weighed 10 g of the crude powder of plant material for 18-40 h. The solvents like water, methanol, chloroform and acetone were used for extraction. The extraction was conducted for 6-8 h/d and finally all the extracts were evaporated under vacuum. The water, methanol, chloroform and acetone extracts of rhizome of the plant were prepared according to standard methods (Harbone, 1998). Nitrogen gas was purged through these extracts to prevent oxidation of secondary metabolites. These extracts were sealed in airtight containers and stored at -40C.
Agar- well diffusion method
Principle
The antimicrobials present in the plant extract are allowed to diffuse out into the medium and interact in a plate freshly seeded with the test organisms. The resulting zones of inhibition will be uniformly circular as there will be a confluent lawn of growth. The diameter of zone of inhibition can be measured in millimetres
Requirements
The anti-bacterial and anti-fungal activity of the plant extracts in solvents viz. methanol, acetone and chloroform in water was assayed against bacterial and fungal cultures.
Plant extracts in aqueous, methanol, acetone and chloroform
Nutrient Agar
Potato Dextrose Agar
Standard antibiotic: Streptomycin
Bacterial cultures: Staphylococcus aureus, Escherichia coli and Salmonella typhi.
Fungal culture: Aspergillus terreus
DMSO (dimethyl sulfoxide)

Procedure
Petriplates containing 20ml Nutrient agar medium were seeded with 24hr culture of Staphylococcus aureus, Escherichia coli, and Salmonella typhi. Similarly, Petri plates containing potato dextrose agar (PDA) seeded with Aspergillus terreus were prepared. Wells were cut and 25 µl of the plant extracts (methanol, acetone and chloroform) were added. All the plates were kept in refrigerator for half an hour for proper diffusion of plant extracts. The plates were then incubated at 37°C for 24 hours. The antibacterial activity was assayed by measuring the diameter of the inhibition zone formed around the well. Streptomycin disc was used as a positive control.
Result and Discussion
All extracts of rhizome of Kaempferia scaposa were screened in-vitro for their anti-microbial activities against clinically isolated bacterial and fungal strains, such as Staphylococcus aureus, Salmonella typhi, Escherichia coli and Aspergillus terreus (Fig. 3-7).
In our results acetone extract showed 10mm zone of inhibition in Salmonella typhi, 2 mm in Staphylococcus aureus, 6mm in Escherichia coli. There were no activity or zone of inhibition was seen against Aspergillus terreus. Chloroform extract showed 2 mm zone of inhibition in Salmonella typhi and 2mm in Escherichia coli. Methanolic extract showed 12mm zone of inhibition against Salmonella typhi and 14mm against Staphylococcus aureus. There was no zone of inhibition against Escherichia coli(Table-1). There were no zone of inhibition was seen against Aspergillus terreus and water extract against all the microorganisms suggested that the selected microorganisms are resistant to them. The plant rhizomes of Kaempferia galanga shown good secondary metabolite products and medicinal activities have not been widely recognized. The entire plant K. galanga were used for antifungal and antibacterial properties for extracting plant metabolites against fungi and bacteria using agar diffusion and the zones of inhibition methods (Kaladhar, et al,2014)
The concentration of the contents in the extract was not known, although the methanolic extracts showed anti-microbial activity suggested that with the minimum inhibition concentration of the contents in acetone and methanolic extracts might be considered as anti-microbial agents

Table no: 1. Anti-microbial activity of rhizome extracts of Kaempferia scaposa
Sample
Extract
Diameter of zone of inhibition in mm


Salmonella typhi
Staphylococcus aureus
Escherichia coli
Aspergillus terreus
Kaempferia scaposa
Acetone
10.0
2.0
6.0
-

Chloroform
2.0
-
2.0
-

Methanol
12.0
14.0
-
-

Water
-
-
-
-



Fig: 3 J : CE – Chloroform extract zone of inhibition of Fig:4 J :CE – Chloroform extract zone of inhibition of Kaempferia scaposa Kaempferia scaposa against Escherichia coli against Salmonella typhi

Fig:5 J :ME – Methanolic extract zone of inhibition of Fig:6J :AE – Acetone extract zone of inhibition of Kaempferia scaposa Kaempferia scaposa against Salmonella typhi against Salmonella typhi

Fig:7 J :ME - Methanolic extract zone of inhibition of Kaempferia scaposa against Staphylococcus aureus
Conclusion
The anti-microbial screening of rhizome extracts against the clinically isolated microorganisms like Salmonella typhi, Staphylococcus aureus, Escherichia coli, Aspergillus terreus was evaluated. Methanolic extracts showed prominent zone of inhibition followed by acetone and chloroform. The poor zone of inhibition is may be due to quantity of bioactive compounds in extracts. Even though it showed medicinal potential of the bioactive compound presents in Kaempferia scaposa.
References
Arambewela, L.S.R., Perera, A. and Wijesundera R.L.C. (1999). Antibacterial activity of Kaempferia galanga. Fitoterapia. 70 (4): 425-427.
Babu, C. R. (1977). Herbaceous flora of Dheradun, C.S.I.R., New Delhi, India.
Bamer, C. J. (1916). Plants of Punjab, Superintendent, Government Printing, Lahore, Pakistan.
Cooke,T. (1903). The Flora of the Presidency of Bombay, I-III, Sri Gouranga Press Pvt. Ltd. Calcutta, India.
Duthie, J. F. (1911). Flora of the upper gangetic plain and of the adjacent Siwalik and Sub-Himalayan tracts. Government Printing Press, Calcutta, India
Fischer, C.E.C. (1921). Flora of Anamalai Hills: Records of the Botanical Survey of India, IX, (I), Superintendent, Government Printing Press, Calcutta, India.
Harborne, J. B. (1998). Phytochemical methods. 3rd ed. London, U. K.: Thomson Science: 44–46.
Hooker, J. D. (1882). The Flora of British India, J. of A.O.A.C. 32 : 291-294.
Mathew, K. N. (1981-1983). The flora of Tamilnadu, Carnatic III, part III. Monocotyledons. The Ranipat Herberium, Tiruchirapalli, India.
Narasinga Rao V, DSVGK Kaladhar (2014). Antioxidant and antimicrobial activities of rhizome extracts of Kaempferia galanga. World Journal of Pharmacy and Pharmaceutial Sciences.3(5) :1180-1189.
Sivarajan V. V. and Indira B. (1994). Oxford and IBH Publishing Co. Pvt. Ltd, New Delhi, India, Ayurvedic drugs and their plant sources: 570.
Ubramani, S.P., Goraya, G.S.(2003).( Some folklore medicinal plants of Kolli hills: Record of a Watti vaidyas Sammelan). J Econ Taxon Bot. 27: 665–78.
Ugemuge, N. R. (1986). Flora of Nagpur District, Maharashtra. Shree Prakashan, Shankarnagar, Nagpur, India.
Warrier, P. K., Nambiar, V. P. and Ramankutty, C. (1993-1995). Madras: Orient Longman Ltd; Ind. Med. Plants: 1–5.
Yadav, S. R. and Sardesai, M. M. (2010). Flora of Kolhapur district. Shivaji University, Kolhapur, India.