Antibacterial and Antifungal Activity of Wattakaka volubilis

 

Sangavi R., Anuradha R.

PG and Research Department of Biochemistry, Sengamala Thayaar Educational Trust Women’s College, Sundarakkottai, Mannargudi-614 016. Tiruvarur (Dt), Tamil Nadu, South India.

*Corresponding Author E-mail:

 

ABSTRACT:

Wattakaka volubilis is a tall woody climber belonging to the family Asclepiadaceae. Leaves, flowers and the rind of unripe fruits are boiled and eaten as a vegetable or used in curries, the cooking removes the bitterness. The ethanolic extracts of Wattakaka volubilis was examined for in vitro antimicrobial activity. The antimicrobial activity was performed on the extracts of Wattakaka volubilis against human pathogenic bacteria such as E.coli, S.aureus and fungal strain such as A.niger, A.flavus. The results were compared with reference drug ampicillin. The extracts exhibited the growth inhibitory activity in a dose dependent manner.

 

KEYWORDS: Wattakaka volubilis, antibacterial activity, antifungal activity.

 

 

INTRODUCTION:

Plants have always been a common source of medicaments, either in the form of traditional preparations or as pure active principles. Medicinal plants represent a rich source of   antimicrobial agents and are used as a source of many potential and powerful drugs in several countries (Srivastava et al., 1996). Plants are known to produce certain bioactive molecules which react with other organisms in the environment, inhibiting bacterial or fungal growth (Chopra et al., 1992) and have a great potential for producing new drugs for human benefit. Traditional medicine prepared using plants contain a vast array of substance that can be used to treat chronic and infectious disease. With an increasing acceptance of traditional medicine as an alternative form of health care, the screening of medicinal plants for active compounds is very important. Traditional medicine based on plants has played a key role in the health care system of many countries like India, China etc. About 60% of the total global population remains dependent on traditional medicines for their health care system3.

Infectious disease casued by bacteria, fungi, viruses and parasities are still a major threat to public health, despite the tremendous progress in human medicine. Pathogenic bacteria have always been considered as a major cause of morbidity and mortality in humans. Even though pharmaceutical companies have produced a number of new antibacterial in the last years, resistance to these drugs has increased and has now become a global concern1. In addition, in developing countries, synthetic drugs are not only expensive and inadequate for the treatment of disease but also often with adulterations and side effects. Therefore, there is need to search new infection-fighting strategies to control microbial infections6. Among the potential sources of new agents, plants have long been investigated.  Because they contain many bioactive compounds that can be of interest in therapeutic. Because of their low toxicity, there is a long tradition of infectious disease in folk medicine. The search is ongoing for new antimicrobial agents, either by the design and synthesis of new agents, or through the search of natural source for as yet  undiscovered antimicrobial  agents, or through the search of natural sources for as yet  undiscovered antimicrobial agents.

 

Medicinal plants are a valuable natural resource and regarded as potentially safe drugs. They have been playing an important role in alleviating human sufferings by contributing herbal medicines in the primary health care systems of rural and remote hilly areas where more than 70% of population depends on folklore and traditional system of medicines. Medicinal plants have been tested for biological and antimicrobial activity5.

 

The plant Wattakaka volubilis  is a climbing shrub of the family Asclepiadaceae. Traditionally, the plant is useful in cold and eye disease. Leaves are used as an application to boils abscesses. The study scientifically validates the plant in traditional medicine6. The GC-MS analysis of ethanol extract of Wattakaka volubilis showed Fifteen compounds were identified. The pharmacological activities such as antioxidant activity9, antimicrobial activity5, antidiabetic activity antioxidant activity8, Silver nanoparticle Synthesis9.cytotoxic activity8. hepatoprotective activity8.

 

The objectives of the present study is to study the in vitro antimicrobial activity of ethanolic extract of Wattakaka volubilis against bacterial species such as Escherichia coli, Staphylococcus aureus and fungal species like Aspergillus niger. Aspergilus flavus by disc diffusion method and compare the activity Ampicillin (bacterial species) and (fungal species) as standard drug.

 

MATERIALS AND METHODS:

Plant Collection of Plant Materials:

The fresh leaves of Wattakaka volubilis were collected from Tiruchirapalli. The leaves were washed thoroughly with tap water, shade dried, homogenized to fine powder.

 

Preparation of Plant Powder:

The plants were air dried under shade for 10-15 days. Then the dried materials were grinded to fine powder using an electric grinder and stored in air tight bottles. The powder matter was used further phytochemical analysis, in vitro antimicrobial activity.

 

Preliminary Phytochemical Screening:

Qualitative phytochemical analysis was cartied out for all the extracts as per the standared methods (Kokate et al., 1995).

 

Preparation of Media:

Muller Hinton agar Media for Bacteria.

 

Antimicrobial Assay:

Disc Preparation:

The 6mm (diameter) discs were prepared from Whatmann No. 1 filter paper. The discs were sterilized by autoclave at 121°C.After the sterilization the moisture discs were dried on hot air oven at 50°C.Then various solvent extract discs and control discs were prepared.  

Collection of test Microorganisms:

The Bacterial strains of Escherichia coli and Staphylococcus aureus and the fungal strains of Aspergillus niger were obtained from Microbial Type culture Collection Centre (MTCC), Chandigarh.

 

Assay of Antibacterial Activity:

Antibacterial activity test was carried out following the modification of the method originally described by Bauer et al., (1966). Muller Hinton agar was prepared and autoclaved at 15 lbs pressure for 20 minutes and cooled to 45ºC. The cooled media was poured on to sterile petriplates and allowed for solidification. The plates with media were seeded with the respective microbial suspension using sterile swab. The various solvents extract prepared discs individually were placed on the each petriplates and also placed control and standard (Gentamicin (10 µg) for Bacteria) discs. The plates were incubated at 37ºC for 24 hrs. After incubation period, the diameter of the zone formed around the paper disc were measured and expressed in mm.

 

Assay of Antifungal Activity:

Antifungal activity test was carried out following the modification of the method originally described by Bauer et al., (1966). Potato Dextrose Agar (PDA) was prepared and autoclaved at 15 lbs pressure for 20 minutes and cooled to 45ºC. The cooled media was added 10ml/L tartaric acid (10%) act as antibacterial agents and poured on to sterile petriplates and allowed for solidification. The plates with media were seeded with the respective microbial suspension using sterile swab. The various solvents extract prepared discs individually were placed on the each petriplates and also placed control and standard (Ketoconazole (10 µg)) discs. The plates were incubated at 28ºC for 72 hrs. After incubation period, the diameter of the zone formed around the paper disc were measured and expressed in mm.

 

RESULTS AND DISCUSSION:

Qualitative Phytochemical Analysis:

Qualitative Phytochemical analysis of ethanolic extract of Wattakaka volubilis represented in Table 1. In the present study, the investigation of phytochemical screening of ethanolic extracts of leaves of Wattakaka volubilis. The qualitative phytochemical analysis revealed the presence of alkaloids, saponin, phytosterols, phenols tannins, flavonoid, steroids triterpenes and coumarins were present Carbohydrates, glycosides, protein and amino acid are absent. Phytochemical constituents such as tannins, flavonoids and several aromatic compounds or secondary metabolites of plants serve as defense mechanism against predation by many microorganisms. The curative properties of medicinal plants are perhaps due to the presence of phenolic compounds, saponins and phytosterols. The presence of alkaloids, saponin, flavonoids, phenolic compounds, tannins phytosterol and terbenoids are used in analgesic, and antipllasmodic and bacteriocidal activities.

 

 

Table 1: Phytochemical screening of Wattakaka volubilis

S. No

Name of the test

Ethanol extract

1

Carbohydrate

-

2

Tannins

+

3

Saponins

-

4

Flavonoids

+

5

Alkaloids

+

6

Quinones

-

7

Glycosides

-

8

Cardiac glycosides

-

9

Terpenoids

-

10

Triterpenoids

-

11

Phenols

+

12

Coumarins

-

13

Steroids and phytosteroids

+

14

Phlobatannins

-

15

Anthraquinones

-

 

Plate 1: Antibacterial and antifungal activity of ethanolic extract of Wattakaka volubilis

 

 

Antimicrobial Activity:

The antimicrobial activity of ethanolic extracts of Wattakaka volubilis were studied in different concentrations (50 mg/ml, 100 mg/ml, 200 mg/ml, 500 mg/ml, 1000 mg/ml) against two pathogenic bacterial strains (Escherichia coli, Staphylococcus aureus) and two fungal strains (Aspergilus niger, Aspergilus flavus). Antibacterial and antifungal potential of ethanolic extracts were assessed in terms of zone of inhibition of bacterial growth. The results of the antibacterial activities are presented in table 2 and table 3. The antibacterial and antifungal activity of the extracts increased in concentration of extracts (mg/ml) as compared with standard drug such as Ampicillin for bacteria and fungi. The results revealed that the bacterial species, Staphylococcus aureus was more sensitive as compared to Escherichia coli and fungal species Aspergillus flavus shows good result as compared to Aspergillus niger. (Plate 1, Table 2).

 

The inhibitory effect of Wattakaka volubilis plant ethanolic extracts showed at 50, 100, 200, 500, 1000 mg/ml were (13, 14, 14.3, 14.3, 1.15, 16.3mm) Escherichia coli,(6.3, 6.6, 6.3, 7.3, 24mm) Staphylococcus aureus for bacterial strains (Plate 1),  and were (6, 6.3, 6.3, 7, 6.6, 14.3mm) Aspergillus niger (6.3, 8.3, 8, 11, 21, 10.3mm) Aspergillus flavus for fungal strains(Plate 2). The results shows that Wattakaka volubilis extracts were found to be more effective against all the microbes tested. In the present study, ethanolic extracts obtained from Wattakaka volubilis plant shows significant activity against most of the tested bacterial and fungal strains. The results were compared with standard antibiotic drugs Ampicillin for fungi.

 

 

The results showed that Wattakaka volubilis were found to be more effective against all the microbes tested. In the present study, ethanolic extracts obtained from Wattakaka volubilis showed significant activity against bacterial and fungal strains. The results of the present study showed that ethanol extracts of Wattakaka volubilis leaves has potent anti microbial activities. Thus the ethanolic extracts of leaves of Wattakaka volubilis may be attributed to the presence of phenolic compounds and flavonoids etc., Therefore, further investigation is needed to isolate and identify the active compounds present in the extract and its efficacy. However, detailed study is required to find out the specific bioactive compounds responsible for antimicrobial property through various advanced techniques.

 

 

 

 

Table 2: Antibacterial activity and antifungal activity  of ethanolic extract of Wattakaka volubilis.

S.

No

Species

Zone of Inhibition (mm in diameter)

Plant extract (µg/ml)

Standard antibiotic

(Ampicillin)

50 (µg/ml)

100 (µg/ml)

200 (µg/ml)

500 (µg/ml)

1000 (µg/ml)

1.

Escherichia coli

13 ± 1.42

14 ± 1.42

14.3±1.25

14.3±2.36

1.15±0.82

16.3±0.47

2.

Staphylococcus aureus

6.3±0.47

6.6±0.47

6.6±0.47

6.3±0.47

7.3±0.47

24±0.82

3.

Aspergillus Niger

6±0.23

6.3±0.47

6.3±0.47

7±0.82

6.6±0.47

14.3±0.47

 

 

 

 

ACKNOWLEDGEMENT:

Authors and thankful to Dr. V. Dhivagaran, M.Sc., D. C. M., Ph.D., Correspondent, S.T.E.T Women’s college, sundarakkottai, mannargudi for his support and guidance.

 

REFFERENCES:

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2.     Chopra, R.N., Nayer, S.L., Chopra, I.C. 1992. Glossary of Indian medicinal plants, 3rd edn. Council of scientific and Industrial Research, New Delhi.7-246.

3.     Kumar, M., Sridevi, K.N.M., Nanduri, S., Rajagopal, S. 2004. Anticancer and immune stimulatory compounds from Andrographis paniculata. J. Ethanopharmacol., 92:291-295.

4.     Rajeswari Satapathy, Ram S Jadhav and Paramjyoti L Swamy. 2016. Antioxidant activity of  Wattakaka volubilis (Linn.f) leaf extract in carbon tetrachloride induced mice, Indian journal of Experimental Biology 54:467-471

5.     Ramachandran A, M. Senthilkumar and D. Vinoth kumar, Evaluation of Antimicrobial Activity of Leaf Extracts of Wattakaka volubilis (L.) Stapf, A. Ramachandran et.al, Int. Journal of Engineering Research and applications 4 (1): 06-10

6.     Shahla Najafi, 2011, Studies on Wattakaka volubilis (L.f.) Stap. A medicinally important plant, research journal of pharmaceutical, Biological and chemical Science 2(2): 164-169.

7.     Sieradzki. K., Wu, S. W. and Tomasz, A. 1999. Inactivation of the methicillin resistance gene mecA in vancomycin-resistant Staphylococcus aureus. Micro. Drug Resist. 5(4): 253- 257.

8.     Usha Rani, S., R. Anuradha 2016. Study of Wattakaka volubilis on Tissue Lipids and Antioxidants in Aluminium Sulphate Exposed Rats, Int. J. Pharm. Sci. Rev. Res., 36(2):A.No 33: 203-208.

9.     Waseem iqbal Khanday, Eyini Muthukumarasamy and Balaji Paulraj 2016. Characterization and evaluation of biological activities of silver nano particles using Wattakaka volubilis Linn.F. Int J Pharm Bio Sci Oct; 7(4): 117 – 123.

 

 

 

 

Received on 13.04.2017                              Modified on 04.07.2017

Accepted on 26.07.2017                             © RJPT All right reserved

Research J. Pharm. and Tech 2017; 10(11): 3775-3778.

DOI: 10.5958/0974-360X.2017.00828.9