Anti-candidal activity of homoeopathic drugs: An in-vitro evaluation

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Anti­candidal activity of homoeopathic drugs: An in­vitro evaluation
Gupta G, Srivastava A K, Gupta N, Gupta G, Mishra S. Anti­candidal activity of homoeopathic drugs: An in­vitro
evaluation.Indian J Res Homoeopathy 2015;9:79­85

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 invitro
anti­candidal effect of few homoeopathic drugs in 30 and 200 potencies against human pathogenic Candida
albicans under in­vitro conditions and compared with standard antifungal drug ketoconazole (control), rectified spirit
(control/vehicle) and distilled water (vehicle) by «DQ»inhibition zone technique«DQ».

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 «DQ»evidence based medicine«DQ» depicting that homoeopathic medicines not only work in invivo
but are equally effective in in­vitro conditions having definite inhibitory activity against Candida albicans.
How to cite this article:

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, sub­acute and chronic localized or
opportunistic systemic infection in human beings. [1],[2],[3] 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. [4],[5],[6]
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 muco­cutaneous regions, viz. oral cavity, rectum vagina etc. [7],[8],[9],[10] Oral swabs or rinses are positive for
C. albicans in up to 40% of healthy adult subjects, while 20­25% 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. Long­term 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 anti­Candida 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 hospita l ­acquired (nosocomial) Candida infections may behave like
minor epidemics with selection of more virulent strains [11] 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 mixed­species communities. [12] 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 [13],[14],[15],[16],[17],[18],[19],[20],[21],[22],[23],[24],[25] and humans. [26],
[27],[28],[29],[30],[31],[32] However, there seems to be few reports on in­vitro 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 yeast­like cells and fungal mycelium. For isolation, rest part of swab was
inoculated in petri dishes 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 (3­6 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 pre­prepared petridishes poured with SDA.
In­vitro Inhibitory Effect of Homoeopathic Drugs Against Human Pathogenic Fungi
Disc method was used to assess the in­vitro anti­candidal 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 in­vitro conditions and compared with standard antifungal drug
ketoconazole (control), rectified spirit (control/vehicle) and distilled water (vehicle). Testing was done by ‘inhibition zone
technique.’ [33],[34]{Table 1}{Table 2}
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.
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}{Figure 2}
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 bulk­form 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
bulk­form source material to exert a ‘usual’ pharmacological dose­response effect. In typical clinical pharmacology,
lower bulk­form “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.
[35],[36] 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 [37] as in modern nanotechnology methods of microfluidization, [38],[39] sonication [40],[41] and
vortexing. [42] The combined impact of these mechanical nanosizing procedures [41] would be able to modify the
properties of the remedy, [43],[44],[45] generating remedy source nanoparticles, [46],[47] as well as Silica crystals and
amorphous nanoparticles. [45],[47],[48]
The results of these in­vitro 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 in­vivo but are
equally effective in in­vitro 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 [46],[47] suggesting that nanoparticles are different from bulk­form materials as a function of
their small size, including acquired adsorptive, [49],[50] electromagnetic, optical, thermal, and quantum properties. [51],
[52],[53],[54],[55] However, the mode of inhibitory action of homoeopathic medicines is a matter of further research and
need of the hour.
The results achieved in these in­vitro 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 side­effects 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 photo­plates and
composing the paper for publication.
Financial Support and Sponsorship
Conflict of Interest
There are no conflict of interest.
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