RESEARCH ARTICLE


Antimicrobial Evaluation of Leaf and Stem Extract of Cordia macleodii



Deepkumar Joshi1, *, Rinku Patel2, Nupur Patel2, Devendra Patel1, Chintan Pandya2
1 Chemistry Division, Sheth M. N. Science College, Patan;
2 Chemistry Department, HVHP Institute of PG Studies and Research, Kadi


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© Joshi et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Chemistry Division, Sheth M. N. Science College, Patan; Tel: +91-9974147411; E-mail: deepjoshi359@yahoo.co.in,


Abstract

The leaf and stem of cordia macleodii plant were investigated for evaluating antibacterial and antifungal properties.Different extracts of leaf and stem viz. double distilled water, methanol, ethyl acetate ane n-hexane were collectedand content was optimized for the maximum amount of extraction with varying time of 6 and 12 h. The extracts weretested for their antimicrobial activities against gram-positive bacteria (B. sutilis), gram-negative bacteria (E. coli) andfungi (A. niger). The extracts were found to be more active against bacteria as compared to fungi. It was also observedthat the antibacterial and antifungal potential of the water extracts were found to be excellent as 1A (38 mm) and 1B (22mm) compared to the other extracts.

Graphical Abstract:

Keywords: Antibacterial and antifungal evaluation, antimicrobial, Cordia macleodii, leaf extract, medicinal plants, stem extract.



INTRODUCTION

Medicinal plants are found to have great potential in healing the wound with minimal discomfort and pain to the patient [1]. Cordia macleodii belonging to Boraginaceae family and familiarly known as Dahiphalas, has green leaves and broad ovate. The leaves of cordia macleodii are found to possess antimicrobial property as reported by Acharya et al. [2]. The flavonoids content present in the leaf of the plant cordia macleodii is reported to exhibit antioxidant and hepatoprotective activities [3]. Also, the HPTLC profile conducted for analyzing the leaf content of the plant, shows the presence of alkaloids, glycosides, tannins at 254 and 366 nm [4]. The biological properties of the solvent extracts of leaf and other part of cordia macleodii have attracted us to study the antimicrobial properties against bacterial and fungal strains.

EXPERIMENTAL

Materials and Methods

Cordia macleodii plant was identified and collected for the analysis by the authentic authority from the Junagadh region of Gujarat state in India. Stem bark and leaves of the plant were cleaned to remove the dust and washed with water and dried. The dried parts were crushed to powder and stored in dry place away from moisture and direct heat. Solvents required for extraction were purchased from Merck Ltd. and SD fine chemicals.

Extraction

Stem and leaf powders around 12 gm were added to 300 ml of solvents (double distilled water, methanol, ethyl acetate and n-hexane) and were exposed to hot extraction method for 6h using Soxhlet method of extraction [5] and 12h by heating at 80°C with constant stirring [6]. The filtration process was carried out using whatman filter paper no. 1, avoiding the loss of solvent. The left over residues were dried and weigh to calculate the extractable matter in various solvents. The crude extract (filtrate) was collected and used for further biological evaluation.

The solvent extracts collected from stem and leaf were numbered in order to identify the samples during further analysis. The water extract collected after 6 h was titled 1A and that after 12 h was named 1B. Similarly, the solvent extracts from methanol, ethylacetate and n-hexane were named 2, 3 and 4, respectively. Notation ‘A’ was used for 6h and ‘B’ for 12h extract for each solvent extract as indicated in Table 1.

Table 1.

Annotation of extracts collected from stem and leaf of Cordia macleodii.


Time / Extract Double Distilled Water Methanol Ethylacetate n-hexane
6h 1A 2A 3A 4A
12h 1B 2B 3B 4B

Antibacterial and Antifungal Evaluation

The antimicrobial tests were carried out using well-diffusion method with standard protocols. Each gram-positive, gram-negative bacteria and fungi were employed for testing the antimicrobial potential of the stem and leaf extracts of the plant. B. subtilis (ATCC no. 6633-gram-positive bacteria), E. coli (ATCC no. 25922-gram-negative bacteria) and A. niger (ATCC no. 1015-fungi) were used for antibacterial and antifungal testing of the compounds. Zone of inhibition (mm) was observed for each of the sample ranging from 1A to 4B and the results were reported in the Tables 2 and 3. Antibiotics like ciprofloxacin and fluconazole available in market were used as standard for antibacterial and antifungal analyses respectively.

Table 2.

Antibacterial and antifungal evaluation of different extracts from leaf of Cordial macleodii.


Extract Gram-positive bacteria
B. subtilis
Gram-negative bacteria
B. subtilis
Fungi
A. niger
1A 22 0 0
1B 38 0 15
2A 6 0 16
2B 8 0 0
3A 7 0 0
3B 9 0 5
4A 7 0 0
4B 7 0 0
Std. 25 24 -
Std. - - 25

Values indicated in the table above are Zone of inhibition (mm)

Std. drug for antibacterial activity is ciprofloxacin and antifungal activity is fluconazole

Table 3.

Antibacterial and antifungal evaluation of different extracts from stem of Cordial macleodii.


Extract Gram-positive bacteria
B. subtilis
Gram-negative bacteria
B. subtilis
Fungi
A. niger
1A 4 0 6
1B 16 0 7
2A 6 0 3
2B 13 0 7
3A 3 0 2
3B 7 8 3
4A 3 2 6
4B 19 5 9
Std. 25 24 -
Std. - - 25

Values indicated in the table above are Zone of inhibition (mm)

Std. drug for antibacterial activity is ciprofloxacin and antifungal activity is fluconazole

RESULTS AND DISCUSSION

Antimicrobial Activity

The antimicrobial results showed in Tables 2 and 3 exhibited promising results against gram-positive bacteria B. subtilis and fungi A. niger, but showed poor results against gram-negative bacteria E. coli, when compared with the standards ciprofloxacin and fluconazole.

Antibacterial and Antifungal Results of Leaf Extracts

From Table 2, it was very clearly observed that none of the extracts was active against gram-negative bacteria E. coli. Water extracts 1A (6h) and 1B (12h) exhibited excellent results (22 and 38 mm) against gram-positive bacteria B. subtilis. Leaf extract 1A was found to be inactive against A. niger, but exhibited moderate zone of inhibition against the same fungal strain. As compared to the standards ciprofloxacin and fluconazole, other extracts exhibited low activity viz. 2A, 2B, 3A, 3B, 4A and 4B against the gram-positive bacterial strain B. subtilis. Methanol extract 2A showed moderate results (15 mm) and ethylacetate extract 3B exhibited poor activity (5 mm) against A. niger

Antibacterial and Antifungal Screening of Stem Extracts

For testing antibacterial and antifungal potentials of the stem extracts as indicated in Table 3, same microbial strains B. subtilis, E. coli and A. niger were utilized. Very few extracts were found to show positive results against gram-negative bacteria E.coli viz. 3B (ethylacetate-8 mm), 4A (n-hexane-2 mm) and 4B (n-hexane-5 mm). All the extracts showed poor to moderate results against B. subtilis (3 mm to 19 mm) as compared to the standard (25 mm). Among those, Water extract 2A and n-hexane extract 4B, exhibited moderate results of 16 mm and 19 mm irrespectively. Stem extracts (1A to 4B) were found to possess low antifungal property when tested against A. niger.

CONCLUSION

The antimicrobial results very well indicated that the water (1B) and n-hexane (4B) extracts collected after 12 hrs were found to show excellent results, also, the antibacterial potential of water extract of leaf (1B) of Cordia macleodii was found to be more than the standard drug ciprofloxacin. Overall, the extracts were active against gram-positive bacteria B. subtilis and fungi A. niger. It was also observed that the extracts collected after 12 hrs exhibited comparatively more antibacterial and antifungal values than the extracts collected after 6 h. Further optimization in the extraction method and identification of the chemical constituent in the leaf and stem extracts can help in providing a newer antimicrobial agent to the society.

CONFLICT OF INTEREST

The authors confirm that this article content has no conflict of interest.

ACKNOWLEDGEMENTS

Authors are thankful to Kadi Sarva Vishwvidyalaya, Gandhinagar and Sarva Vidyalaya Kelavani Mandal, Kadi for providing infrastructural facilities to the researchers to carry out the work. Authors are thankful to Prof. B. A. Golakiya for his kind help in literature survey and identification and collection of Cordia macleodii plant.

REFERENCES

[1] MacKay DJ, Miller AL. Nutritional support for wound heal-ing Alternat Med Rev 2003; 8(4): 359-77.
[2] Bhide B, Ashok BK, Acharya RN, Ravishankar B. Anti-microbial and wound healing activities of Cordia macleodii Hook f & Thoms leaves Ind J Nat Prod Resour 2011; 1: 198-203.
[3] Qureshi NN, Kuchekar BS, Logade NA, Haleem MA. Antiox-idant and hepatoprotective activity of Cordia macleodii leaves Saud Pharm J 2009; 17(4): 299-302.
[4] Bhide B, Acharya RN, Naria P, Pillai APG, Shukla VJ. Phar-macognostic Evaluation of Cordia macleodii Hook Stem Bark Pharmacog J 2011; 3(26): 49-53.
[5] Wang L, Weller CL. Recent advances in extraction of nutraceuticals from plants Trend Food Sci Technol 2006; 17(6): 300-12.
[6] Akroum S, Satta D, Lalaoui K. Antimicrobial, antioxidant, cytotoxic activities and phytochemical screening of some Al-gerian plants Eur J Sci Res 2009; 31(2): 289-95.