|Year : 2015 | Volume
| Issue : 2 | Page : 79-85
Anti-candidal activity of homoeopathic drugs: An in-vitro evaluation
Girish Gupta, AK Srivastava, Naveen Gupta, Gaurang Gupta, Sunil Mishra
Medical Mycology Lab, Gaurang Clinic and Centre for Homoeopathic Research, Lucknow, Uttar Pradesh, India
|Date of Submission||13-Jan-2015|
|Date of Acceptance||05-Jun-2015|
|Date of Web Publication||30-Jun-2015|
Dr. Girish Gupta
Gaurang Clinic and Centre for Homoeopathic Research, B-1/41, Sector A, Opp. Novelty (Aliganj), Kapoorthala, Aliganj, Lucknow - 226 024, Uttar Pradesh
Source of Support: None, Conflict of Interest: None
Background: Candida albicans is an opportunistic pathogenic fungus accounting for up to 75% of all candidal infections in human beings. Generally Candida grow and survive as commensals but slight modification of the host defense system can transform Candida albicans into a pathogen.
Materials and Methods: Samples collected from the oral cavity and tongue of the patients suspected of suffering from oral candidiasis were incubated for growth of Candida. Fermentation and assimilation test confirmed the species as Candida albicans. Disc method was used to assess the in-vitro anti-candidal effect of few homoeopathic drugs in 30 and 200 potencies against human pathogenic Candida albicans under in-vitro conditions and compared with standard antifungal drug ketoconazole (control), rectified spirit (control/vehicle) and distilled water (vehicle) by "inhibition zone technique".
Results: Homeopathic drugs namely Acid benzoicum, Apis mellifica, Kali iodatum, Mezereum, Petroleum, Sulphur, Tellurium, Sulphur iodatum, Graphites, Sepia, Silicea and Thuja occidentalis in 30 and 200 potencies were tested against Candida albicans. Mezereum in 200 and 30 potency showed maximum inhibition of growth of Candida albicans followed by Kali iodatum 200 while Kali iodatum 30 and Petroleum 30 had minimum inhibition.
Conclusion: The results of these experiments support the concept of "evidence based medicine" depicting that homoeopathic medicines not only work in in-vivo but are equally effective in in-vitro conditions having definite inhibitory activity against Candida albicans.
Keywords: Antifungal, Candida albicans, homoeopathic drugs, In-vitro inhibitory activity
|How to cite this article:|
Gupta G, Srivastava A K, Gupta N, Gupta G, Mishra S. Anti-candidal activity of homoeopathic drugs: An in-vitro evaluation. Indian J Res Homoeopathy 2015;9:79-85
|How to cite this URL:|
Gupta G, Srivastava A K, Gupta N, Gupta G, Mishra S. Anti-candidal activity of homoeopathic drugs: An in-vitro evaluation. Indian J Res Homoeopathy [serial online] 2015 [cited 2020 Feb 20];9:79-85. Available from: http://www.ijrh.org/text.asp?2015/9/2/79/159522
| Introduction|| |
Fungal infections are of great significance in tropical countries like India where heat and humidity provide conditions favorable for onset, growth and persistence of fungus. Candida albicans is an opportunistic pathogenic fungus found as a part of normal microflora in human digestive tract causing episodic, acute, sub-acute and chronic localized or opportunistic systemic infection in human beings. ,, Adherence, perspiration, dimorphism and/or germ tube formation, phenotype switching, interference with host defense mechanism, hormonal imbalance, synergism with bacteria and production of hydrolases from the metabolites have been identified as factors enhancing the virulence of C. albicans. ,,
Candida albicans is the most important species in the genus Candida and accounts for up to 75% of all candidal infections. In general, innate and acquired host defense mechanisms act in concert with the resident bacterial flora such that Candida organisms grow and survive as commensals. Even a slight modification of the host defense system, or host ecological environment, can assist the transformation of C. albicans into a pathogen capable of causing infections that may be lethal. The most common body sites showing asymptomatic colonization by Candida are the alimentary tract and muco-cutaneous regions, viz. oral cavity, rectum vagina etc. ,,, Oral swabs or rinses are positive for C. albicans in up to 40% of healthy adult subjects, while 20-25% of healthy women carry C. albicans in the vagina. Colonization by Candida is thought to occur at an early age with the organisms being acquired during passage through the birth canal, during nursing or from food. Long-term colonization is probably responsible for eliciting the circulating Immunoglobulin G and mucosal secretory Immunoglobulin A antibodies to C. albicans that are detectable in most healthy individuals. It is these acquired host responses in conjunction with the anti-Candida activities of polymorphonuclear leukocytes and macrophages that probably play a significant part in normally restricting C. albicans to superficial growth at mucosal sites.
Although recent evidence suggests that some hospital-acquired (nosocomial) Candida infections may behave like minor epidemics with selection of more virulent strains  it is often the commensal (endogenous) organisms that are believed to be the initial sources of infection. However, it is important to recognize that C. albicans have the ability to live in harmony with the host, for a lifetime, within the resident complex microflora present on mucosal surfaces. In the oral cavity, C. albicans grows and survives by competing and cooperating with an estimated 300 or more species of bacteria. There is compelling evidence that C. albicans and C. dubliniensis form tight associations with specific oral bacterial species and that these promote adhesion and colonization by mixed-species communities.  Thus, when Candida infections arise, they often occur in association with bacteria. On the other hand, there is also strong evidence to suggest that components of the resident microflora, present in the oral cavity and at other mucosal sites, perform to check C. albicans growth. That is why factors that perturb the normal microflora, such as antibiotic therapy, or changes in hormonal or mucosal secretions, may encourage C. albicans overgrowth.
A considerable number of experimental studies have been conducted to test the efficacy of homoeopathic drugs against fungal and viral diseases of plants, animals ,,,,,,,,,,,, and humans. ,,,,,, However, there seems to be few reports on in-vitro antimycotic effect of homoeopathic drugs against human pathogenic fungi in general and C. albicans in particular. The present study was, therefore, undertaken to determine the inhibitory effect of various homoeopathic drugs against human pathogenic fungi confirming the biological activity of homoeopathic drugs in higher dilutions.
To determine the inhibitory effect of various homoeopathic drugs against human pathogenic fungus C. albicans confirming the biological activity of potentized drugs in higher dilutions.
| Materials and Methods|| |
Isolation of Human Pathogenic Candida albicans
The samples were collected from the oral cavity and tongue of the patients suffering from oral candidiasis who presented themselves at Gaurang Clinic and Centre for Homoeopathic Research for treatment. Part of the oral swab was examined directly in Potassium hydroxide (KOH) (10%) slide mount for the presence of yeast cells. KOH preparation of swab showed fair number of yeast-like cells and fungal mycelium. For isolation, rest part of swab was inoculated in Petri dish More Detailses poured with Sabouraud's Dextrose Agar (SDA)-Emmons modified (HI Media B. No. 9039) incubated at 37°C ± 1°C for 72 hours. Microscopic examination of 4 days old culture showed globose, short, ovoid sometimes elongated blastoconidia (3-6 μm) on corn meal agar. Reynold's braude phenomenon was observed by incubating blastoconidia in human serum at 37°C and germination were found to be more than 70%.
Fermentation and assimilation test further confirmed the identity of the species as C. albicans. Oral swab from healthy persons was kept as control. However, for checking contamination, if any, petri dishes poured with SDA in four replicates were exposed to the environment, gave several mycelial fungi dominated by species of Aspergillus but there were no C. albicans in the working environment.
From the colonies of C. albicans obtained, solitary colony of C. albicans was taken with the help of loop, inoculated in a test tube containing 5 ml Sabouraud's dextrose broth (Test tube 1) and incubated at 37°C ± 1°C for 24 hours. Now 1 ml of this broth containing C. albicans was taken and added in another test tube containing 4 ml of plain Sabouraud's dextrose broth (Test tube 2). Similarly, 1 ml of this broth containing C. albicans was taken and added in another test tube containing 4 ml of plain Sabouraud's dextrose broth (Test tube 3). The same procedure was repeated and Test tube 4 and 5 were prepared. Now this Test tube 5 was kept at room temperature for 24 hours. Broth from Test tube 5 was taken with the help of cotton swab stick and plated on pre-prepared petridishes poured with SDA.
In-vitro Inhibitory Effect of Homoeopathic Drugs Against Human Pathogenic Fungi
Disc method was used to assess the in-vitro anti-candidal effect of homoeopathic drugs [Table 1] and [Table 2] against human pathogenic C. albicans. Homoeopathic drugs namely Acid benzoicum, Apis mellifica, Kali iodatum, Mezereum, Petroleum, Sulphur, Tellurium, Sulphur iodatum, Graphites, Sepia, Silicea and Thuja occidentalis in 30C and 200C potencies were tested against C. albicans under in-vitro conditions and compared with standard antifungal drug ketoconazole (control), rectified spirit (control/vehicle) and distilled water (vehicle). Testing was done by 'inhibition zone technique.' ,
|Table 1: In-vitro inhibitory effect of homoeopathic drugs in 30C potency against C. albicans by "inhibition zone technique" (diameter of disc= 12 mm) |
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|Table 2: In-vitro inhibitory effect of homoeopathic drugs in 200C potency against C. albicans by "inhibition zone technique" (diameter of disc= 12 mm) |
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A volume of 20 ml sterilized SDA was plated on 27 sterilized petridishes and allowed to solidify. 1 ml SDA medium was seeded with the culture broth, mixed well and poured over the surface of all the petri dishes already plated with the medium. Discs (12 mm in diameter) of sterilized Whatman No. 1 filter paper were dipped in different homoeopathic drug potencies, standard antifungal drug ketoconazole ([100 μg/ml]/disc) and rectified spirit (control/vehicle) were placed on the center of each petridish separately.
Petridishes were then incubated at 37°C ± 1°C for 72 hours. Inhibition of growth or no growth of C. albicans indicated the effectiveness of homoeopathic drugs in different potencies. Ketoconazole rectified spirit and distilled water were used for comparison as controls. The experiments were repeated 3 times and the mean effective area of the zone of inhibition was calculated.
| Results|| |
Different potencies of Mezereum, Kali iodatum, Acid benzoicum, Petroleum, Sulphur iodatum, Sulphur and Tellurium [Table 1] and [Table 2] and [Figure 1] and [Figure 2] have variable inhibitory effect against C. albicans. Among different drug potencies, Mezereum in 200C and 30C showed maximum inhibition of growth of C. albicans followed by Kali iodatum 200C while Kali iodatum 30C and Petroleum 30C had minimum. However, ketoconazole showed maximum inhibition. The difference in inhibition of different drug formulations may be ascribed to variations in drug compositions and medicament.
|Figure 1: In-vitro anti-candidal activity of homoeopathic drugs in 30 potency|
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|Figure 2: In-vitro anti-candidal activity of homoeopathic drugs in 200 potency|
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| Discussion|| |
Before the inception of nanoparticle, it was believed that homoeopathic medicines do not have any traces of the original drug substance in dilutions beyond '24 X' or '12 C' potency because repeated dilution steps leave progressively fewer and fewer molecules of bulk-form source material in a true solution, until eventually none should persist in solution diluted past Avogadro's number (6.023 × 10 23 ), that is, potencies higher than '24X' or '12C'. As a result, conventional medical scientists and chemists rejected the plausibility of Homoeopathy because of the presumptive lack of sufficient bulk-form source material to exert a 'usual' pharmacological dose-response effect. In typical clinical pharmacology, lower bulk-form "doses" should exert lesser effects, but contrary to this, the trituration and succussion procedures used in the preparation of homoeopathic remedies generates nanoparticles of the source material. Trituration with mortar and pestle is a manual method for mechanical grinding or milling, similar to ball milling used in modern nanotechnology. , Similarly, manual succussions introduce intense turbulence, particle collisions, and shear forces into solution that break off smaller and smaller particles of remedy source material as well as Silica from the walls of the glass containers or vials  as in modern nanotechnology methods of microfluidization, , sonication , and vortexing.  The combined impact of these mechanical nanosizing procedures  would be able to modify the properties of the remedy, ,, generating remedy source nanoparticles, , as well as Silica crystals and amorphous nanoparticles. ,,
The results of these in-vitro studies are highly encouraging and have shown that homoeopathic drugs have definite inhibitory activity against C. albicans in culture plate confirming that these medicines not only work in-vivo but are equally effective in in-vitro conditions. The results of these experiments in culture plate have proved that though the homoeopathic medicines are biologically active as persistent remedy source nanoparticles have been demonstrated with high resolution types of electron microscopy in metal and plant homoeopathic remedies prepared both below and above Avogadro's number , suggesting that nanoparticles are different from bulk-form materials as a function of their small size, including acquired adsorptive, , electromagnetic, optical, thermal, and quantum properties. ,,,, However, the mode of inhibitory action of homoeopathic medicines is a matter of further research and need of the hour.
| Conclusion|| |
The results achieved in these in-vitro experiments support the concept of the 'evidence based medicine' clearly depicting that homoeopathic drugs have definite inhibitory activity against C. albicans. No doubt the effect is less than that of ketoconazole, but ketoconazole being known to cause many side-effects there is need to test more homoeopathic drugs to find out effective homoeopathic drugs against C. albicans and various other human pathogenic fungi. Further studies may open new vistas even in the treatment of resistant strains as well.
These results will definitely clear the misconceptions that Homoeopathy is a placebo therapy and it will have to be accepted that if modern medicine is the science of today Homoeopathy is the science of future.
The authors acknowledge the effort of Mr. Pawan Mishra, a computer programmer for scanning of photo-plates and composing the paper for publication.
Financial Support and Sponsorship
Conflict of Interest
There are no conflict of interest.
| References|| |
Gupta G, Srivastava AK. Cutaneous candidiasis due to adhesive bindi. Biol Mem 2001;27:79-81.
Mukerji KG, Srivastava AK, Singh KP, Garg KL, editors. Advances in Medical Mycology. Vol. 1. New Delhi: Aditya Books Private Limited; 1992. p. 227.
Emmons W, Binford CH, Utz JP, Kwonchung KJ. Medical Mycology. Philadelphia: Lea and Febiger; 1997. p. 592.
Ashman RB, Fulurija A, Apadimitriou JM. Pathogenesis of Candida Albicans
Infection, Advances in Medical Mycology. Vol. 2. New Delhi: Aditya Books Private Limited; 1997. p. 1-28.
Srivastava AK, Singh KP, Ray PK. Protein A induced protection against experimental candidiasis in mice. Mycopathologia 1997;138:21-8.
Srivastava OP, Srivastava AK, Shukla PK, editors. Advances in Medical Mycology. Vol. 2. Lucknow: Council for Advances in Bioresearch and Evoker Research Perfecting Co.; 1997. p. 188.
Marples MJ, editor. The Ecology of the Human Skin. Springfield, IL: Charles C Thomas; 1965. p. 250.
Marples MJ, Somerville DA. The oral and cutaneous distribution of Candida albicans
and other yeasts in Rarotonga, Cook Islands. Trans R Soc Trop Med Hyg 1968;62:256-62.
Ghoniem NA, Refai M. Incidence of Candida
species in Damietta cheese. Mykosen 1968;11:295-8.
Do Carmo-Sousa L. Distribution of yeasts in nature. In: Rose AH, Harrison JS, editors. The Yeasts. Vol. 1. London: Academic Press; 1969. p. 79-105.
Marco F, Lockhart SR, Pfaller MA, Pujol C, Rangel-Frausto MS, Wiblin T, et al.
Elucidating the origins of nosocomial infections with Candida albicans
by DNA fingerprinting with the complex probe Ca3. J Clin Microbiol 1999;37:2817-28.
Lamont RJ, Jenkinson HF. Adhesion as an ecological determinant in the oral cavity. In: Kuramitsu HK, Ellen RP, edirors. Oral Bacterial Ecology: The Molecular Basis. Wymondham, United Kingdom: Horizon Scientific Press; 2000. p. 131-68.
Verma HN, Verma GS, Verma VK, Krishna R, Srivastava KM. Homoeopathic and pharmacopoeial drugs and inhibitors of tobacco mosaic virus. Indian Phytopathol 1969;22:188-93.
Khurana SM. Effect of homoeopathic drugs on plant viruses. Planta Med 1971;20:142-6.
Khurana SM. Chemotherapeutic Potential of Homoeopathic Drugs against Plant Viruses. Proceedings 225 th
Hahnemann Birthday and Annual Function, HMAI, Lucknow; 11 th
April, 1980. p. 29-35.
Abidi SM, Srivastava KM, Gupta RP, Singh BP. Effect of some medicinal drugs on distortion ring spot virus of papaya. New Botanist 1977;4:13-4.
Goswami N, Das D. Possibilities of homoeopathic treatment in plant and animal diseases. Hahnemannian Gleanings 1980;47:332-41.
Khanna KK, Chandra S. Effect of some homoeopathic drugs on the spore germination of four isolates of Alternaria alternate
. Indian Phytopathol 1976;29:195-7.
Khanna KK, Chandra S. A homoeopathic drug (Lycopodium
) controls mango fruit rot caused by Pestalotia mangiferae
Henn. Hahnemannian Gleanings 1980;XLVII: 441-8.
Khanna KK, Chandra S. Homoeopathic drug induced abnormalities in the germ tubes of Pestalotia mangiferae
and Gloeosporium psidii
. Adv Homoeopath 1981;1:35-6.
Singh BP, Gupta GC, Srivastava KM. Homoeopathic drugs as inhibitors of tobacco mosaic virus. Hahnemannian Gleanings 1980;XLVII: 265-7.
Singh BP, Gupta G. Effect of homoeopathic drugs on the growth of Alternaria tenuis
act. and Curvularia lunata
(Wakker) Boedijin, the common leaf spot pathogens of ornamental and cultivated plants. Hahnemannian Gleanings 1981;XLVII: 411-3.
Verma HN. Protection against viruses by homoeo drugs and plants used in Indian medicine system. Adv Homoeopath 1981;1:69-73.
Khurana KK, Gupta G. Homoeopathy: Promise and prospects for plant protection. Adv Homoeopath 1981;1:107-16.
Singh LM, Gupta G. Antiviral efficacy of homoeopathic drugs against animal viruses. Br Homoeopath J 1985;74:168-74.
Gupta G, Garg KL, Chandra B. Effects of homoeopathic drugs against fungi isolated from human patients. Asian Homoeopath J 1995;5:15-8.
Gupta G, Srivastava AK, Chandra B, Gupta N. Cutaneous candidiasis: Therapeutic potential of homoeopathic drugs. Asian Homoeopath J 1997a; 7:8-32.
Gupta G, Srivastava AK, Chandra B, Gupta N. Mycoses: An update over clinical cure with homoeopathic drugs. Asian Homoeopath J 1997b; 7:67-96.
Gupta G, Chandra B. In-vitro
inhibitory effect of some homoeopathic drugs against Candida albicans
. Biol Mem 2003;29:98-101.
Gupta G, Chandra B. A clinico-mycological study on oral candidiasis cases in response to homoeopathic treatment. Homoeopath Herit 2004;20:5-12.
Gupta G. Inhibitory effect of homoeopathic drugs against Candida albicans
study, published in souvenir of Volume 2, December 2004 of Bhartiya Vaigyanik evam Audyogik Anusandhan Patrika (BVAAP) published during Tritiya Akhil Bharitya Vigyaan Sammeelan (3 rd
ABVS) held at Delhi from February 19-21; 2004. p. 305-7.
Gupta G, Chandra B. Inhibitory effect of homoeopathic drugs against Candida albicans
study. Homoeopathy 2004;5:41-3.
Leben C, Kiett GW. A bioassay for tetramethylthiuram disulphide. Phytopathology 1950;40:950-4.
Nene YL, Thapliyal PN. Fungicides in Plant Disease Control. New Delhi, India: Oxford and IBH Publishing Co.; 1979. p. 507.
DeCastro CL, Mitchell BS. Nanoparticles from mechanical attrition. In: Baraton MI, Valencia CA, editors. Synthesis, Functionalization, and Surface Treatment of Nanoparticles. CCRH, New Delhi: American Scientific Publisher; 2002. p. 1-15.
Merisko-Liversidge E, Liversidge GG. Nanosizing for oral and parenteral drug delivery: A perspective on formulating poorly-water soluble compounds using wet media milling technology. Adv Drug Deliv Rev 2011;63:427-40.
Bhattacharyya SS, Mandal SK, Biswas R, Paul S, Pathak S, Boujedaini N, et al. In vitro
studies demonstrate anticancer activity of an alkaloid of the plant Gelsemium
sempervirens. Exp Biol Med (Maywood) 2008;233:1591-601.
Keck CM, Muller RH. Drug nanocrystals of poorly soluble drugs produced by high pressure homogenization. Eur J Pharm Biopharm 2006;62:3-16.
Liu G, Zhang D, Jiao Y, Zheng D, Liu Y, Duan C, et al
. Comparison of different methods for preparation of a stable riccardin D formulation via nano-technology. Int J Pharm 2012;422:516-22.
Ruan B, Jacobi M. Ultrasonication effects on thermal and rheological properties of carbon nanotube suspensions. Nanoscale Res Lett 2012;7:127.
Tang C, Zhou T, Yang J, Zhang Q, Chen F, Fu Q, et al
. Wet-grinding assisted ultrasonic dispersion of pristine multi-walled carbon nanotubes (MWCNTs) in chitosan solution. Colloids Surf B Biointerfaces 2011;86:189-97.
Chan HK, Kwok PC. Production methods for nanodrug particles using the bottom-up approach. Adv Drug Deliv Rev 2011;63:406-16.
Elia V, Napoli E, Germano R. The ′memory of water′: An almost deciphered enigma. Dissipative structures in extremely dilute aqueous solutions. Homeopathy 2007;96:163-9.
Rey L. Can low-temperature thermo-luminescence cast light on the nature of ultra-high dilutions? Homeopathy 2007;96:170-4.
Demangeat JL. NMR relaxation evidence for solute-induced nanosized superstructures in ultramolecular aqueous dilutions of silica-lactose. J Mol Liq 2010;155:71-9.
Chikramane PS, Suresh AK, Bellare JR, Kane SG. Extreme homeopathic dilutions retain starting materials: A nanoparticulate perspective. Homeopathy 2010;99:231-42.
Upadhyay RP, Nayak C. Homeopathy emerging as nanomedicine. Int J High Dilution Res 2011;10:299-310.
Ives JA, Moffett JR, Arun P, Lam D, Todorov TI, Brothers AB, et al
. Enzyme stabilization by glass-derived silicates in glass-exposed aqueous solutions. Homeopathy 2010;99:15-24.
Liu L, Randolph TW, Carpenter JF. Particles shed from syringe filters and their effects on agitation-induced protein aggregation. J Pharm Sci 2012;101:2952-9.
Chi EY, Weickmann J, Carpenter JF, Manning MC, Randolph TW. Heterogeneous nucleation-controlled particulate formation of recombinant human platelet-activating factor acetyl hydrolase in pharmaceutical formulation. J Pharm Sci 2005;94:256-74.
Montagnier L, Aissa J, Ferris S, Montagnier JL, Lavallee C. Electromagnetic signals are produced by aqueous nanostructures derived from bacterial DNA sequences. Interdiscip Sci Comput Life Sci 2009;1:81-90.
Roduner E. Size matters: Why nanomaterials are different. Chem Soc Rev 2006;35:583-92.
Buzea C, Pacheco II, Robbie K. Nanomaterials and nanoparticles: Sources and toxicity. Biointerphases 2007;2:MR17-71.
Yao P, Hughes S. Macroscopic entanglement and violation of Bell′s inequalities between two spatially separated quantum dots in a planar photonic crystal system. Opt Express 2009;17:11505-14.
Iris RB, Mary K. A model for homeopathic remedy effects: Low dose nanoparticles, allostatic cross-adaptation, and time-dependent sensitization in a complex adaptive system. BMC Complement Alternat Med 2012;12:191. Available from: http://www.biomedcentral.com/1472-6882/12/191
[Figure 1], [Figure 2]
[Table 1], [Table 2]