Antifungal activity of selected medicinal herbs against canine dermatophytosis

Abstract

The current study was conducted to evaluate the clinical efficacy of the polyherbal combination containing Acalypha indica, Curcuma longa and common salt for the treatment canine dermatophytosis. Qualitative phytochemical screening was carried out to assess the bioactive components present in extracts of Acalypha indica and Curcuma longa. Three different polyherbal combinations were prepared in different ratios and evaluated for its efficacy. The clinical evaluation was carried out in dogs with dermatophytosis brought to Veterinary Clinical Complex, Veterinary College and Research Institute, Orathanadu. The phytochemical screening of the selected herbs revealed the presence of phytoconstituents viz. alkaloids, flavonoids, glycosides, phenols, tannins, terpenoids, and carbohydrate compounds. The clinical recovery in fungal dermatophytosis in pet animals with an application of polyherbal combination in the ratio of 7:2:1 comprising Acalypha indica, Curcuma longa and saturated salt solution for four times a day for 10 days indicated that topical therapy not only eliminates fungal infection but also control the skin problem without any side effects. Also, the polyherbal antifungal combination will be an efficient and economically affordable formulation in the field to treat dermatophyte infection in pet animals.

Full text

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Journal of Entomology and Zoology Studies 2020; 8(6): 1618-1622
E-ISSN: 2320-7078
P-ISSN: 2349-6800
www.entomoljournal.com
JEZS 2020; 8(6): 1618-1622
© 2020 JEZS
Received: 15-08-2020
Accepted: 07-10-2020
K Vijayakaran
Department of Veterinary
Pharmacology and Toxicology,
Veterinary College and Research
Institute, Tamil Nadu
Veterinary and Animal Sciences
University, Orathanadu, Tamil
Nadu, India
V Ranganathan
Department of Veterinary
Pharmacology and Toxicology,
Veterinary College and Research
Institute, Tamil Nadu
Veterinary and Animal Sciences
University, Orathanadu, Tamil
Nadu, India
P Senthil Kumar
Department of Veterinary
Pharmacology and Toxicology,
Veterinary College and Research
Institute, Tamil Nadu
Veterinary and Animal Sciences
University, Orathanadu, Tamil
Nadu, India
Corresponding Author:
K Vijayakaran
Department of Veterinary
Pharmacology and Toxicology,
Veterinary College and Research
Institute, Tamil Nadu
Veterinary and Animal Sciences
University, Orathanadu, Tamil
Nadu, India
Antifungal activity of selected medicinal herbs
against canine dermatophytosis
K Vijayakaran, V Ranganathan and P Senthil Kumar
DOI: https://doi.org/10.22271/j.ento.2020.v8.i6v.8051
Abstract
The current study was conducted to evaluate the clinical efficacy of the polyherbal combination
containing Acalypha indica, Curcuma longa and common salt for the treatment canine dermatophytosis.
Qualitative phytochemical screening was carried out to assess the bioactive components present in
extracts of Acalypha indica and Curcuma longa. Three different polyherbal combinations were prepared
in different ratios and evaluated for its efficacy. The clinical evaluation was carried out in dogs with
dermatophytosis brought to Veterinary Clinical Complex, Veterinary College and Research Institute,
Orathanadu. The phytochemical screening of the selected herbs revealed the presence of
phytoconstituents viz. alkaloids, flavonoids, glycosides, phenols, tannins, terpenoids, and carbohydrate
compounds. The clinical recovery in fungal dermatophytosis in pet animals with an application of
polyherbal combination in the ratio of 7:2:1 comprising Acalypha indica, Curcuma longa and saturated
salt solution for four times a day for 10 days indicated that topical therapy not only eliminates fungal
infection but also control the skin problem without any side effects. Also, the polyherbal antifungal
combination will be an efficient and economically affordable formulation in the field to treat
dermatophyte infection in pet animals.
Keywords: Dermatophytosis, Herbal drugs, Acalypha indica, Curcuma longa, common salt
Introduction
Dermatophytosis in companion animals is a skin disease caused by a superficial fungal
infection of keratinized skin structures by zoophilic, geophilic or anthropophilic fungal
organisms, most commonly Microsporum canis, M. gypseum and Trichophyton
mentagrophytes [17, 22]. These fungi digest the keratin protein complex in the skin by
keratinases and others enzymes that allows the dermatophyte to burrow deeper into the stratum
corneum in the host and elicit an inflammatory reaction [14, 15]. Because of the pleomorphic
presentation of clinical signs, its infectious and contagious nature, and zoonotic potential,
dermatophytosis is an important disease in small animal medicine. Treatment is recommended
with the goal of shortening the course of the disease to prevent spread to other animals and
human being [22].
The incidence of dermatophytes in animals especially in dogs has been high and serious
problem today. This is because the antifungal drug resistance of the pathogen and the side
effect exhibited by the drugs used to cure fungal diseases. Hence there is great demand for an
efficient, alternative and safer treatment. Recently, there has been increasing interest in the use
of various herbal materials as an alternative medicine to treat some of the fungal infections and
many compounds of natural products have been specifically targeted against pathogenic fungi.
Medicinal herbs could be an alternate source for the treatment of canine dermatophytosis.
Herbal medicines are safe and generally free from side effects. Another important reason for
this revival is that the effectiveness of many traditional medicines is now an accepted fact.
With the above background, the current study has been taken to evaluate the antifungal action
of certain herbal plants against canine dermatophytosis. Based on the literature survey and
traditional use,the following herbs having the antifungal action had been selected, viz.
Acalypha indica [1, 7, 26], Curcuma longa [4, 24, 36], sodium chloride (common salt) [10] and
evaluated for the antidermatophytic activity.
Materials and Methods
Selection of Materials
Medicinal plants with antifungal effects like Acalypha indica, Curcuma longa and common
salt were selected for this study.

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Collection of plant materials
Fresh leaves of Acalypha indica and rhizome of Curcuma
longa were collected from Herbal garden at Veterinary
College and Research Institute, Orathanadu, Tamilnadu,
India. The taxonomic identities of plants were confirmed by
Botanical Survey of India, Coimbatore, Tamilnadu, India. The
collected plants were washed with running tap water, air
dried, homogenized to a fine powder and stored in air-tight
container.
Preparation of crude extracts for phytochemical analysis
About 30 g of dried powder material of Acalypha indica and
Curcuma longa were extracted with 100 ml of distilled water
and methanol respectively using soxhlet apparatus. The
extract was then concentrated using Rotary Evaporator (Buchi
Rotavapor R-300) and stored at 4°C until use.
Estimation of yield percentage
The extraction percentage of extracts of Acalypha indica and
Curcuma longa was calculated by using the following
formula:
Phytochemical screening assay
Qualitative phytochemical screening was carried out to assess
the bioactive components present in extracts of Acalypha
indica and Curcuma longa as described by Trease and Evans
(1989) [35].
Preparation of polyherbal combination
The plant leaves were washed twice thoroughly with sterile
distilled water in order to remove traces of dust and soil
particles and the fresh leaves of about 75 g of Acalypha indica
were taken and ground thoroughly using mixer grinder by
adding 100 ml of water (75%) and the ground leaves mixture
was filtered using Whatman filter paper No 1 and the filtrate
was collected. The turmeric powder was prepared by grinding
the dry turmeric rhizome using mixer grinder. Fifty percent
turmeric extract was prepared by adding 50 gram of turmeric
powder to 100 ml distilled water and keeping it in a magnetic
stirrer rotating at 400 rpm at 50 degree celsius. Saturated salt
solution (SSS) was prepared by adding enough quantity of
salt to distilled water until it gets precipitated.
The different combinations of crude extracts of selected plants
and saturated salt solution (SSS) were tried to find out the
most effective combination against canine dermatophytes.
The crude extracts of plants were taken in different ratio
randomly and the clinical trials were carried out for all the
combinations and it is presented in the Table 1. The most
efficient combination was then determined by comparing the
results of clinical trials. The combination was mixed
thoroughly and ready for use in animals.
Table 1: Different ratio of polyherbal combination
Combination
Acalypha indica
Curcuma longa
SSS
I
5
4
1
II
6
3
1
III
7
2
1
Experimental design
The study was carried out in dogs with dermatophytosis
brought to Veterinary Clinical Complex, VCRI, Orathanadu.
Dogs with clinical signs of pruritus with regular and circular
alopecia, scales and crust with erythematous margin,
hyperpigmentation and a thin desquamation were selected.
The Wood’s lamp test was performed on dogs with the
clinical signs of dermatophytes to establish the tentative
diagnosis of dermatophytosis in dogs. The animals with the
clinical signs of dermatophytosis that were brought to
Veterinary Clinical Complex, VCRI, Orathanadu were
divided into four treatment groups with six animals in each
group and it is presented in the Table 2. Group I was treated
with topical Ketaconazole which serves as the positive
control. Group II, III and IV were treated with polyherbal
combination I, II and III respectively.
Table 2: Experimental design
Group
Number of animals
Treatment
Route of administration
Group I
6
Treated with topical Ketaconazole
Topical application, Four times daily on affected area for 10 days
Group II
6
Combination I
Topical application, Four times daily on affected area for 10 days
Group III
6
Combination II
Topical application, Four times daily on affected area for 10 days
Group IV
6
Combination III
Topical application, Four times daily on affected area for 10 days
The clinical efficacy was assessed by topically applying the
polyherbal combination on the dogs with clinical signs of
dermatophytosis. A score card was prepared with selected
clinical signs viz. alopecia, scales and crust, erythema,
hyperpigmentation, thin desquamation for assessing the
clinical efficacy of polyherbal combination.
Results
Plant extraction yield percentage
The ethnobotanical data of the selected plants and their
extract percentage yield are given in Table 3. The percentage
yield of plant extract obtained from Acalypha indica and
Curcuma longa were (28.09%±0.19) and (14.51%±0.23)
respectively.
Table 3: Ethnobotanical data of selected plants and their extract yield percentage
Plant
Family
Local name
Plant part used
Extract yield (%) (±SD)
Acalypha indica
Euphorbiaceae
Kuppaimeni
Leaves
28.09% ±0.19
Curcuma longa
Zingiberaceae
Turmeric, Manjal
Rhizome
14.51%±0.23
Qualitative phytochemical screening
The result of phytochemical screening of extracts of Acalypha
indica and Curcuma longa are presented in Table 4. The
preliminary screening for the phytochemicals showed the
presence of phytoconstituents viz. alkaloids, flavonoids,
glycosides, phenols, tannins, terpenoids, and carbohydrate
compounds.

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Table 4: Qualitative phytochemical screening of extracts
S. No
Phytochemicals
Extract of Curcuma longa
Extract of Acalypha indica
Polyherbal combination
1
Alkaloids
Positive
Positive
Positive
2
Glycosides
Positive
Positive
Positive
4
Phenols
Positive
Negative
Positive
5
Tannins
Negative
Positive
Positive
6
Phlobatannins
Negative
Negative
Negative
8
Flavonoids
Positive
Positive
Positive
9
Terpenoids
Positive
Positive
Positive
10
Saponins
Negative
Negative
Negative
11
Carbohydrates
Positive
Positive
Positive
12
Steroids
Positive
Positive
Positive
Assessment of efficacy of polyherbal formulation
Clinical evaluation was done in the dogs having
dermatophytosis that are brought to the Veterinary Clinical
Complex, Orathanadu. The combination was used four times
daily for 10 days. The dogs recovered from the clinical
lesions in 10-15 days. The efficacy of the polyherbal
combination was assessed with the score card given in Table
5.
Table 5: Score card analysis for clinical evaluation (after 15 days)
Clinical signs
Before treatment (0 day)
Positive control
Combination I
Combination II
Combination III
Alopecia
Present
Absent
Present
Absent
Absent
Scales and crust
Present
Absent
Present
Absent
Absent
Erythema
Present
Absent
Absent
Absent
Absent
Hyperpigmentation
Present
Absent
Present
Present
Absent
Thin desquamation
Present
Absent
Present
Absent
Absent
Woods lamp examination
Positive
Negative
Positive
Negative
Negative
Out of the three combinations prepared combination III
comprising Acalypha indica, Curcuma longa and SSS with
the ratio of 7:2:1 was found to be effective against canine
dermatophytosis when compared to the other two
combinations. In alopecic areas growths of new hairs with
visible improvement in the clinical signs were noticed and
Woods lamp examination also found to be negative. The
results were comparable with that of the positive control
(Group I).
Discussion
Dermatophytosis is a specific fungal disease of the epidermal
tissues in the skin, caused by keratinophilic and keratinolytic
genera such as Microsporum, Trichophyton and
Epidermophyton. It is an endemic infection in many countries
throughout the world affecting companion animals (dogs,
cats), domestic animals (calves), and laboratory animals
(rabbits) as well as humans [23, 30]. The most frequently found
dermatophytes infestation in dogs are Microsporum spp. and
Trichophyton spp. Some dermatophytes are zoonotically
important which infect primarily the animals and are
transmitted from infected animals to human beings on many
occasions [25]. In dogs, nearly 70% of cases are caused by
Microsporum canis, 20% by M. gypseum, and 10% by
Trichophyton mentagrophytes [11]. Dermatophytes are
significant due to their zoonotic potential and the concern of
owner of pets with some time severe inflammatory diseases.
The antifungals commonly used in systemic treatment of
dermatophytosis in dogs and cats include itraconazole,
terbinafine and griseofulvin [19]. The quick development of
resistance and rapid global spread of resistant fungi demands
to find new antifungal agents. Currently emergence of
antifungal resistant clinical isolates leads to failure in the
treatment of mycosis [2]. Keeping this in view, the current
study had been carried out to identify the therapeutic potential
of certain herbal plants for their antifungal activity
particularly against dermatophytes.
Acalypha indica commonly known as kuppaimeni in Tamil
and Curcuma longa (turmeric) were identified to have
antifungal property which is in accordance several researchers
[1, 7, 26].
Acalypha indica belongs to the family Euphorbiaceae, is used
to treat superficial fungal infections [1]. The plants belonging
to Euphorbiaceae family contains high concentration of
flavonoids, phenols and alkaloids. The presence of these
phytochemicals may be responsible for the potent antifungal
activity [26]. Oksana et al. [27] reported that flavonoids
(quercetin, kaempferol, isorhamnetin, isoquercitrin), phenolic
derivatives (gallicin, gallic, syringic, and caffeic acids), and
coumarin (scopoletin) containing plants have potent anti-
fungal activity against Microsporum spp. and Trichophyton
spp. Dinesh kumar et al. [16] reported that leaves and flowers
of Acalypha indica contain flavonoids kaempherol,
glycosides, clitorin, mauritianin and nictiflorin.
Acalypha indica leaf juice is added to oil or lime to treat a
variety of skin disorders and other ailments [33]. Azmahani et
al. [7] reported the leaves and roots of the aqueous extract of
Acalypha indica inhibit the growth of Microsporum canis.
Curcuma longa belongs to Zingiberaceae family, possesses
powerful antifungal activity, as demonstrated in many studies
[4, 24, 36].
Curcumin, demethoxy curcumin and bis-demetho-
hydroxycurcumin, are three pharmacologically important
Curcuminoids that have been isolated from Curcuma longa
[20]. They have been shown to possess anti-oxidant, anti-
inflammatory, anti-carcinogenic, anti-mutagenic, anti-fungal,
anti-viral and anti-cancer properties [12]. The therapeutic
properties of C. longa include insecticidal [3], antimicrobial
[18], antifungal [21], antimalarial, antiviral and antioxidant
properties. Hu et al. [21] reported that the inhibition behaviour
of C. longa on fungal growth (Aspergillus flavus) is involved
in its ability to disrupt the integrity of plasma membrane and
mitochondrial dysfunction, inducing metabolic stagnation.
Chen et al. [13] reported that the ethanol extract of C. longa
can disrupt the synthesis of critical proteins and enzymes,
which may ultimately inhibit the growth of fungi (Fusarium

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graminareum). The antifungal effects were found to be
related to the disruption of fungal cell membrane systems,
specifically the inhibition of ergosterol synthesis and the
respiratory chain.
Sodium chloride salt is a known antifungal agent that acts by
altering osmotic gradients, forcing organisms to expend
energy in osmoregulation, diverting it away from growth at
concentrations beyond tolerance limits, the osmoregulatory
processes are overloaded and death occurs [10].
In the present study, the polyherbal combination containing
Acalypha indica, Curcuma longa and sodium chloride is
found to be clinically effective against dermatophytes in pet
animals. The results of the current study are in accordance
with the reports of the previous studies of antidermatophytic
activity of the herbal plants selected. Leaves and roots of the
aqueous extract of Acalypha indica inhibit the growth of
Microsporum canis [7]. Somchit et al. [32] reported that the
non-polar extract of Acalypha indica showed antifungal
action and at 30 mg/mL chloroform extract, the activity was
statistically similar to the antifungal drug ketoconazole.
Sudhakar Chekuri et al. [33] reported that, methanol, ethanol
and acetone shade dried extract of Acalypha indica leaf
extract showed wide range of antibacterial and antifungal
activity and can be used and administered in medical practice.
Herbal drug preparation containing rhizome powder cured
ringworm infection caused by Trychophyton verrucosum in
12 cattles and Microsporum canis in 21 dogs within12-15
days of treatment [31]. The crude methanol extract of C. longa
has inhibitory effect against some clinical isolates of
dermatophytes. It was demonstrated that 18-month-old and
freshly distilled oil isolated from rhizome of C. longa showed
the most potent antifungal effect against 29 clinical isolates of
dermatophytes with MIC values of 7.2 and 7.8mg/mL,
respectively. Trichophyton rubrum, T. mentagrophytes,
Epidermophyton floccosum, and Microsporum gypseum were
suppressed by 1: 40–1: 320 dilutions of turmeric oil. An in
vivo study on infected guinea pigs with T. rubrum
demonstrated that dermal application of turmeric oil (dilution
1: 80) induced an improvement in healing of the lesions after
2–5 days and it caused the lesions after 6-7 days of
consumption to vanish [6]. Oil from C. Longa proved to be
more effective against dermatophytes than curcumin.
Vaijayanthimala et al. [36] demonstrated that C.longa rhizome
ethanolic extract showed more activity against T. rubrum
(0.57 mg/ml) than T. mentagrophytes (1.15 mg/ml).
In the current study, the phytochemical analysis of the
aqueous extract of Acalypha indica leaves showed the
presence of alkaloids, glycosides, tannins, terpenoids,
flavonoids and carbohydrates. The results are in accordance
with the phytochemical analysis done earlier in several studies
in which the qualitative phytochemical screening indicated
the presence of alkaloids, phenols, saponins, steroids,
flavonoids and catechol [9, 29]. Takle et al. [34] also stated that
qualitative chemical examinations of various extracts and
fractions of Acalypha indica revealed the presence of
alkaloids, steroids, flavonoids, glycosides, tannins and
carbohydrates. The preliminary phytochemical analysis of the
methanolic extract of turmeric also showed the presence of
tannins, alkaloids, saponins, flavonoids, terpenoids and
cardiac glycosides which is in accordance with the study of
Rajesh et al. [28]
Somchit et al. [32] reported that the non-polar extract of
Acalypha indica leaves especially at 30 mg/mL chloroform
extract, has showed antifungal activity which was as potent as
to the conventional antifungal drug ketaconazole and
fluconazole. Hence, they can be used in treatment of fungal
diseases caused by Microsporum canis and some Candida sp
which is in correlation with the current study. Bhadauria and
Kumar [8] studied that the bound flavonoid extracts as well as
free flavonoid extracts present in certain herbal plants showed
prominent antidermatophytic activity against all the
dermatophytes. The phytochemical studies carried out in the
present study indicate the presence of flavonoids in both
Acalypha indica and Curcuma longa as well as in the
polyherbal formulation. The presence of flavonoids in the
selected herbs and in polyherbal antifungal combination could
be attributed for the antidermatophytic action.
The clinical recovery in fungal dermatophytosis in pet
animals with an application of polyherbal combination for
four times a day for 10 days indicated that topical therapy not
only eliminates fungal infection but also control the skin
problem without any side effects. Also, the polyherbal
antifungal combination will be an efficient and economically
affordable formulation in the field to treat dermatophyte
infection in pet animals.
Conclusion
In conclusion, it is suggested that the application of
polyherbal combination containing Acalypha indica, Curcuma
longa, and sodium chloride in the ratio of 7:2:1 for 10 days
has the potential to reduce the fungal infection in pet animals.
Further studies are warranted to explore the possible
mechanisms of antifungal action behind the selected herbs.
Acknowledgments
The authors are thankful to Tamil Nadu Veterinary and
Animal Sciences University (TANUVAS), Chennai and
TANII Scheme, Tamil Nadu Government.
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