An in-vitro Study on the effect of  Calotropis Gigantea in different  Potencies on Trichophyton Rubrum

Mansoor  Ali KR; Nisha Paul; Kannan  G; Jijith  US; Chandraja  CV

doi:10.48165/ahr.2024.9.4.2

Authors

Mansoor Ali KR PhD Scholar, Vinayaka Mission Research Foundation, Salem. Professor Govt Homoeopathic Medical College Calicut
Nisha Paul Professor, Vinayaka Mission’s Homeopathic Medical College, Salem
Kannan G Professor, Vinayaka Mission’s Kirupananda Variyar Medical College Salem
Jijith US Ph.D. Assistant Professor, College of Pharmaceutical Sciences Govt. Medical College Kozhikode
Chandraja CV Associate Professor, Department of Community Medicine, JIMS Homoeopathic Medical College, Hyderabad.Telangana-509325

DOI:

https://doi.org/10.48165/ahr.2024.9.4.2

Keywords:

Calotropis gigantea, Trichophyton rubrum, Antifungal activity, Minimum Inhibitory Concentration (MIC), Homeopathic medicine

Abstract

Background: The emergence of resistant fungal strains due to irrational use of antifungal agents highlights the need for alternative therapies. Calotropis gigantea, known for its traditional medicinal uses, has been explored for its antifungal properties against Trichophyton rubrum, a common dermatophyte causing skin infections. Materials and Methods: Standard strains of T. rubrum (ATCC MYA-029) were cultured on Sabouraud Dextrose Agar (SDA). Various potencies of C. gigantea (3X, 6X, 3C, 6C, 12C, 30C, 200C, and 1000C) were evaluated for antifungal activity using the disc diffusion method. Griseofulvin served as the positive control, and homeopathic placebo (90% ethanol) as the negative control. Minimum Inhibitory Concentration (MIC) values were determined using turbidometry and analyzed statistically using the Kruskal-Wallis Test. Results: Lower potencies of C. gigantea (3C, 3X, and 12C) demonstrated significant antifungal activity, with inhibition zones ranging from 9 ± 0.5 mm to 10 ± 0.3 mm (p < 0.05). MIC50 and MIC90 values for C. gigantea 3C were 100 µL and 200 µL, respectively, indicating higher efficacy compared to higher potencies (30C, 200C, and 1000C), which showed MIC50 and MIC90 values of 500 µL and 900 µL. Griseofulvin exhibited the highest antifungal activity with MIC50 and MIC90 values of 100 µL each. The negative control showed no significant inhibition. Discussion: The antifungal efficacy of C. gigantea is attributed to its bioactive compounds, including flavonoids and terpenoids. Lower potencies exhibited stronger activity, aligning with traditional applications of the plant. The findings support its potential as a topical antifungal treatment and highlight the value of MIC in guiding effective dosing strategies. Conclusion: Homeopathic medicine Calotropis gigantea demonstrates significant antifungal activity, particularly in lower potencies (3X, 3C, and 12C), offering a promising alternative for managing dermatophytic infections. Further research should focus on optimizing treatment regimens, exploring synergies with other homeopathic medicines, and validating findings in clinical settings to address antifungal resistance effectively.

References

1. World Health Organization. Global report on superficial fungal infections. Geneva: WHO; 2022. 2. Goyal S, Kumar S, Rawat P, Dhaliwal N. Antifungal activity of Calotropis procera towards dermatoplaytes. International Journal of Advances in Pharmacy, Biology and Chemistry. 2013;2(3):2277- 4688.

3. Gupta AK, Lyons DC. The rise and challenge of chronic dermatophytosis: lessons from T. rubrum. Mycopathologia. 2015;180(5-6):387–96.

4. Zhan P, Liu W. Pathogenic fungi in superficial infections. Clin Microbiol Rev. 2017;30(4):942–70. 5. Nenoff P, Krüger C, Schaller M, et al. Mycology – an update. Part 2: dermatomycoses: clinical picture and diagnostics. J Dtsch Dermatol Ges. 2015;13(8):778–803.

6. Havlickova B, Czaika VA, Friedrich M. Epidemiological trends in skin mycoses worldwide. Mycoses. 2008;51 Suppl 4:2–15.

7. Verma S, Madhu R. The great Indian epidemic of superficial dermatophytosis: an appraisal. Indian J Dermatol Venereol Leprol. 2017;83(3):227–36.

8. Rudramurthy SM, Shankarnarayan SA, Dogra S, et al. Recurrent dermatophytosis: a growing concern in India. Clin Microbiol Rev. 2018;31(1):e00047-17.

9. Bauer, A.W., W. M. M. Kirby., J.C., Sherris and M. Turck. 1966. Antibiotic Susceptibility Testing by a Standardized Single Disk Method. Am. J. Clin. Pathol. 45: 493-496.

10. Wiegand I, Hilpert K, Hancock RE. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc. 2008;3(2):163-75.

11. MTCC. Microbial Type Culture Collection & Gene Bank. Institute of Microbial Technology, Chandigarh.

Available from: https://www.imtech.res.in.

12. Larone DH. Medically Important Fungi: A Guide to Identification. 5th ed. ASM Press; 2011.

13. SBL India Pvt. Ltd. Product Catalogue. Available from: https://www.sblglobal.com.

14. Granade TC, Artis WM. Factors affecting griseofulvin susceptibility testing of Trichophyton rubrum in microcultures. J Clin Microbiol. 1982 Dec;16(6):1043-7. doi: 10.1128/jcm.16.6.1043-

1047.1982. PMID: 7161372; PMCID: PMC272536. 15. HiMedia Laboratories. Product Information for Griseofulvin. Available from: https://www. himedialabs.com.

16. National Assessment and Accreditation Council (NAAC). Accreditation details. Available from: https://www.naac.gov.in.

17. CLSI. Clinical and Laboratory Standards Institute. Methods for Antifungal Susceptibility Testing. M38-A2. Wayne, PA: CLSI; 2008.

18. NCCLS. Performance Standards for Antifungal Disk Diffusion Susceptibility Testing of Yeasts. Approved Guideline M44-A. NCCLS, Wayne, PA; 2004.

19. Andrews JM. Determination of minimum inhibitory concentrations. J Antimicrob Chemother. 2001;48 Suppl 1:5-16.

20. Shimadzu Corporation. UV-1900i UV-Vis Spectrophotometer. Product Information. Available from: https://www.shimadzu.com.

21. Pfaller MA, Diekema DJ. Epidemiology of invasive candidiasis: a persistent public health problem. Clin Microbiol Rev. 2007;20(1):133-63.

22. Wiegand I, Hilpert K, Hancock RE. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc. 2008;3(2):163-75.

23. Scholz R, Meinhof W. Susceptibility of Trichophyton rubrum to griseofulvin. Mycoses. 1991 Sep Oct;34(9-10):411-4. doi: 10.1111/j.1439-0507.1991. tb00804.x. PMID: 1820520.

24. PS Kumar, E Suresh, S Kalavathy. Review on a potential herb Calotropis gigantea (L.) R. Br Sch Acad J Pharm. 2013;2:135–43.

25. Santos DD, Barros ME, Hamdan JS. Establishing a method of inoculum preparation for susceptibility testing of Trichophyton rubrum and Trichophyton mentagrophytes. Journal of clinical microbiology. 2006 Jan;44(1):98-101.

26. . Kumar G, Karthic L, Bhaskara Rao KV. In vitro anti candida activity of Calotropis

27. gigentica against clinical isolates of Candida. J Pharm Res. 2010;3:539–42.