Bioscience Discovery, 7(1):25-29, Jan - 2016

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Research Article

Alectra parasitica A. Rich. – An Unexplored Parasitic Plant with

Potential as Antimicrobial Agent
Kakpure MR1* and Rothe SP2

1.Department of Botany, S.M.D. Bharti Mahavidyalaya, Arni Dist. Yavatmal – 445103.

2.Department of Botany, Shri Shivaji College of Arts, Commerce and Science, Akola- 444001.
*Email - manojkakpure@rediffmail.com

Article Info Abstract

Received: 07-11-2015, The present study was carried out to evaluate antimicrobial activity of three different
Revised: 13-12-2015, extracts of plant - Alectra parasitica A. Rich. The samples were tested against 7
Accepted: 20-12-2015 bacterial strains and 2 fungal strains by well diffusion method. The antibacterial
activity was tested against human pathogenic strains i.e. Staphylococcus aureus,
Bacillus subtilis, Escherichia coli, Proteus vulgaris, Salmonella typhi, Pseudomonas
Keywords:
aeroginosa and Streptococcus pneumoniae while, antifungal activity was tested
Alectra parasitica, against Aspergillus niger and Candida albicans. The results showed that, A.
antimicrobial agent, parasitica acetone and ethanol extracts showed significant antimicrobial activity
parasitic plant and well against microorganism tested. The water extract exhibited no antimicrobial activity
diffusion method. against microorganism tested. Comparatively it was showed that, A. parasitica
acetone extract posses more antibacterial as well as antifungal activity than ethanol
extract.

INTRODUCTION it and it has not been known sufficiently to

Microorganisms are causative agents of practitioners of indigenous medicine in other parts
almost all kinds of acute and chronic diseases. The of the country. Properties and uses of this drug as
plant based antimicrobials have enormous known to local people were also recorded. It is felt
therapeutic potential (Robbers et al., 1996; Barbour that this may prove to be a medicinal plant of
et al., 2004; Devi & Lawrence, 2014). Higher plants economic importance (Awasthi et al., 2008; George
have traditionally been used in folk medicine et al., 2011).
showing inhibition against bacteria, fungi and Alectra parasitica A. Rich is effective in the
yeasts (Hulin et al., 1998). Alectra parasitica A. treatment of various infectious diseases.
Rich is an unexplored parasitic plant locally known Kakpure & Rothe (2012) reported the presence
as Nirgunda (family-Scrophulariaceae) parasite on of alkaloids, carbohydrates, sterols, glycosides,
the roots of Vitex negundo L. This plant is saponin, flavonoids, quinone, coumarins and
indigenous to India and first reported by Kamble &
phenolics compounds in A. parasitica which is
Pradhan (1988) from Akola district of Maharashtra.
It has been used in the treatment of leprosy, useful for treating different diseases and
tuberculosis, paralysis, swellings, fever, expulsion infections as having a potential of providing
of intestinal worms and constipation for centuries in useful drugs of human use. Earlier, only Saxena
traditional Ayurvedic medicinal practices, & Vyas (1993) reported different extracts of A.
remaining strictly confined to a limited areas parasitica var. chitrakutensis (Rau.) R. Prasad
(Anonymous, 1986; Chopra et al., 1956; Rangari, possesses antibacterial activity and none of the
2006; Saxena & Saxena 2009; Sikarwar et al., extracts of the samples shows antifungal
2007). Only Vaidoos and mendicants practiced with activity.
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 Kakpure and Rothe
Despite the intense uses of A. parasitica as incubated overnight at 37 ºC for 24 hrs each
medicinal plant and the researches concerning the bacterial strain. Amoxicillin, Gentamycin and
pharmacological importance of this plant, the Chloramphenicol standard antibiotics were used as
thorough knowledge of antimicrobial activity is still a positive reference while, DMSO was used as a
scarce. So, in the present study an attempt has been negative control.
made the laboratory evaluations to assess the For fungi, Sabouraud’s Dextrose Agar for
antimicrobial properties of Alectra parasitica A. Candida albicans and Aspergillus niger were
Rich. prepared in plates as the media. Fungal strain
Candida albicans plates were incubated at 37°C for
MATERIALS AND METHODS 48 hrs while, Aspergillus niger plates were
The antimicrobial activity of Alectra incubated at RT 25-30°C for 72 hrs. Clotrimazole,
parasitica A. Rich in three different extracts i.e. Ketoconazole and Nystatin standard antibiotics
acetone, ethanol and water extracts were carried out were used as a positive reference while, DMSO was
by using well diffusion method described by used as a negative control. The antimicrobial
(Mukharjee, 2002; Satish et. al., 2008 and Nitha et. activities were then assessed by measuring the
al., 2012). diameter of the growth–zone of inhibition in
Collection and identification of plant materials: millimeters (including well diameter of 6 mm) for
The plant Alectra parasitica A. Rich was collected the test organisms comparing to the standard
from Popatkhed, Dhargad, Patur, Shahanur and antibiotics. The diameters of zone of inhibition (in
Shirla forest areas of Akola district, Maharashtra. mm) are presented in results.
The collected plants were identified with the help of
standard floras (Kamble & Pradhan, 1988; Naik, Table-1: Composition of bacterial maintenance
1998; Singh et. al. 2001) and herbarium specimens Medium (Nutrient agar)
were deposited in Herbarium of Department of S. N. Ingredients Quantity
Botany, Shri Shivaji College, Akola and the 1 Peptic digest of animal
5 gm/ L
collected whole plant materials was shade dried and tissue
grinding into a powder, packed in polythene bags 2 Yeast extract 1.5 gm/ L
until further use. 3 Beef extract 1.5 gm/ L
Preparation of extracts: The dried plant powdered 4 Sodium Chloride 10 gm/ L
material of A. parasitica (100 g) was extracted with 5 pH 7.5
acetone, ethanol and water. The flask kept it on 6 Agar-agar 15 gm/ L
rotary- shaker at 150 rpm for 24 hrs. After 24 hours, 7 Distilled water 1000ml
the supernatant was filtered through Whatman filter
paper No. 41. The acetone and ethanol solvent Table-2: Composition of fungal maintenance
suspension was completely evaporated using Medium (SDA)
vacuum while, water suspension was boiled in S. N. Ingredients Quantity
water bath and evaporated. Then, the residues 1 Peptone (Meat & Casein) 10 gm/ L
obtained was dissolved in 1% DMSO4 (Dimethyl 2 Dextrose monohydrate 20 gm/ L
Sulphoxide) and used for further study. 3 Agar- Agar 15 gm/ L
Bacterial cultures: Bacterial cultures were 4 pH 5.8
obtained from Microbial Type Culture Collection 5 Distilled water 1000 ml
(MTCC), Chandigarh, India. All the bacterial
cultures were maintained in nutrient agar and stored RESULTS AND DISCUSSION
at 40C. Naturally occurring substances of plant origin have
Antimicrobial Assay: The antimicrobial assay was been reported to inhibit the growth of
performed by well diffusion method. Nutrient Broth microorganisms. Bacterial infection seems
was prepared in tubes as the media for test bacteria. especially controllable due to good hygiene and the
The bacterial inoculums were spread evenly on the availability of effective antibacterial drugs (Dhale &
surface of the nutrient agar plates using a sterilized Mogle, 2011). In the present study, the
cotton swab. For agar well diffusion method, wells antimicrobial activity of Alectra parasitica A. Rich
were prepared in the plates with the help of a cork- of acetone, ethanol and water extracts were tested
borer (0.6 cm). 100 μl of the test compound was by using well diffusion method against 7 bacterial
introduced into the each well. The plates were and 2 fungal strains.
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 Bioscience Discovery, 7(1):25-29, Jan - 2016
The antibacterial activity was tested against the following human pathogenic strains viz. Staphylococcus
aureus, Bacillus subtilis, Escherichia coli, Proteus vulgaris, Salmonella typhi,

Table 3 - Antibacterial activity of A. parasitica showing zone of inhibition in mm.

Acetone Ethanol Water

S. N. Organisms AMXC GNTM CRMP
extract extract extract
1 S. aureus 16 17 -- 18 18 21
2 Bacillus subtilis 15 12 -- 12 25 28
3 Escherichia coli 07 06 -- 15 24 22
4 Proteus vulgaris 18 13 -- 07 26 23
5 Salmonella typhi 15 13 -- 14 25 23
6 P. aeroginosa 11 09 -- 19 28 24
7 S. pneumoniae 16 14 -- 10 25 23
Where, AMXC = Amoxicillin, GNTM = Gentamycin and CRMP = Chloramphenicol.

Table 4 - Antifungal activity of A. parasitica showing zone of inhibition in mm.

Acetone Ethanol Water

S.N. Organisms CTMZ KTCZ NYST
extract extract extract
1 Aspergillus niger 12 15 -- 10 14 27
2 Candida albicans 18 12 -- 08 18 22
Where, CTMZ = Clotrimazole, KTCZ = Ketoconazole and NYST = Nystatin.

Graph 1 – showing antibacterial activity of A. parasitica showing zone of inhibition in mm.

30
Zone of inhabition in mm

25
20
Acetone
15 Ethanol
10 Water
Amoxicillin
5
Gentamycin
0 Chlorampheni

Pseudomonas aeroginosa and It was found that, acetone and ethanol extracts
Streptococcus pneumoniaeBacterial Strains
while, antifungal showed significant antibacterial as well as
activity was tested against Aspergillus niger and antifungal activity of A. parasitica against the above
Candida albicans. The results were compared with mentioned microorganism. The water extract of A.
the antibacterial activity of three standard parasitica exhibited no antimicrobial activity
antibiotics Amoxicillin, Gentamycin, against microorganism tested (Table-3 & 4). The
Chloramphenicol and antifungal activity of three results showed that, A. parasitica acetone extract
standard antibiotics Clotrimazole, Ketoconazole and posses more antibacterial activity than ethanol
Nystatin respectively. extract.

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 Kakpure and Rothe

Graph 2 – showing antifungal activity of A. parasitica showing zone of inhibition in mm.

m
m
n

n
o

o
Z

n
h

n
e

o
t
f

i
i

i
30
25 Acetone
20
Ethanol
15
10 Water
5
0
Aspergillus niger Candida albicans
Fungal strains

The significant and highest antibacterial activity possesses antibacterial activity and none of the
(zone of inhibition 18 mm) was shown by acetone extracts of the samples shows antifungal activity.
extract of A. parasitica against Proteus vulgaris, This antimicrobial activity is due to the presence of
successively zone of inhibition 16 and 15 mm were different secondary metabolites (aromatic
shown by Staphylococcus aureus, Streptococcus substances) which can synthesize by plants (Borde
pneumonia and Bacillus subtilis, Salmonella typhi et al., 2013; Mogle, 2013). In the present study both
respectively. Whereas, the minimum zone of the acetone and ethanol extracts shows the
inhibition 07 mm was shown by acetone extract significant antifungal activity. This is due to the
against E.coli. However, ethanol extract shown higher solubility of the active compounds into
moderate zone of inhibition 17, 14, 13, 12, 09 & 06 acetone and ethanol solvents (Kakpure & Rothe,
mm against Staphylococcus aureus, Streptococcus 2012; Devi & Lawrence, 2014). So, the result
pneumonia, Proteus vulgaris, Salmonella typhi, obtained in the present study showed that A.
Bacillus subtilis, Pseudomonas aeroginosa and parasitica is effective against several bacterial
E.coli respectively. The highest antibacterial infection and fungal pathogens.
activity (zone of inhibition 28 mm) was shown by The results obtained in the present study are in
standard antibiotic Gentamycin against agreement to a certain degree with the
Pseudomonas aeroginosa and Chloramphenicol traditional uses of the plant. Plant based
against Bacillus subtilis while, least antibacterial antimicrobials have enormous therapeutic
activity (zone of inhibition 07 mm) was shown by potential as they can serve the purpose with
standard antibiotic Amoxicillin against Proteus
lesser side effects that are often associated with
vulgaris. The detailed results are depicted in (table-
3 & graph-1). synthetic antimicrobials. From the above results
The highest antifungal activity (zone of it can be concluded that, A. parasitica whole
inhibition 18 mm) was shown by acetone extract of plant powder extracts have great potential as
A. parasitica against Candida albicans while, the antimicrobial compounds against
least (zone of inhibition 12 mm) was showed by microorganisms and that they can be used in
both acetone and ethanol extract of A. parasitica the treatment of various infectious diseases
against Aspergillus niger and Candida albicans caused by resistant microorganisms. Alectra
respectively. Water extract was not seen the parasitica A.
antifungal activity against tested organisms. The significant antibacterial as well as
highest antifungal activity (zone of inhibition 27 antifungal activity and so this plant may be serve as
mm) was shown by standard antibiotic Nystatin leads for the development of new pharmaceuticals
against Aspergillus niger, while least antifungal that address hither to unmet therapeutic needs.
activity (zone of inhibition 8 mm) was shown by However, further investigation on isolation and
standard antibiotic Clotrimazole against Candida characterization of the active principles of this plant
albicans (table-4 & graph-2). extracts responsible for the antimicrobial activity is
However, the earlier workers Saxena & necessary and it would give a comprehensive
Vyas (1993) reported different extracts of A. evidence of bioactive potential of this medicinal
parasitica var. chitrakutensis (Rau.) R. Prasad plant.
http://jbsd.in 28 ISSN: 2229-3469 (Print)
 Bioscience Discovery, 7(1):25-29, Jan - 2016
The millenarian use of this plant in folk Kakpure MR and Rothe SP, 2012. Phytochemical
medicine suggests that it represents an economic screening of Alectra parasitica A. Rich. – A rare
and safe alternative to treat infectious diseases. parasitic medicinal plant. J. Adv. Res. Pharmac. &
Biol., 2(1):103-111.
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How to Cite this Article:

Kakpure MR and Rothe SP, 2016. Alectra parasitica A. Rich. – An Unexplored Parasitic Plant with
Potential as Antimicrobial Agent. Bioscience Discovery, 7(1):25-29.

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