Received: 29th August-2013 Revised: 12th Sept-2013

Accepted: 16th Sept-2013

Research article
ANTIMICROBIAL ACTIVITY OF ALOE VERA LEAF EXTRACT

Kedarnath1, Kamble Kaveri M2, Vishwanath B Chimkod and C S Patil3

1
Singhania University, Pacheri Bari, Rajasthan
2
Plant Tissue Culture and Genetic Engineering Lab, Department of P. G and Research Studies
in Botany, Gulbarga University, Gulbarga- 585106, Karnataka, INDIA
3
Department of Biotechnology, B.V. Bhoomreddy College of UG and PG Studies, Bidar,
Karnataka, INDIA
E-mail: kaverikamble@gmail.com
ABSTRACT: The antimicrobial activity of aloe vera extract was tested against pathogenic bacteria like
Staphylococcus aureus, Klebisella pneumonia and E.coli and fungi like Aspergillus niger and Candida at a dose of
1:20 mg/ml and 2:40 mg/ml by using cup plate diffusion method. Various solvents such as petroleum ether, chloroform
and methanol were used for extracts. The results reveal that, methanol and petroleum ether at a dose of 20 mg/ml has
showed significant activity against Klebisella pneumonia and E.coli whereas in fungi, methanol extract showed
significant activity against Aspergillus niger and Candida. Methanol extract has showed maximum inhibitory activity
against E.coli and Candida. Petroleum ether has showed moderate inhibitory activity against Klebisella pneumonia and
Candida. The zone of inhibition was measured and compared with standard Gentamycin (1 mg/ml). However, in none
of the above mentioned extracts the inhibition zone was not more than that found in standard i.e., Gentamycin.
Key words: Aloe vera, antimicrobial activity, Gentamycin

INTRODUCTION
Traditional medicine is in practice for many centuries by a substantial proportion of the population of many centuries.
It is recognized that in some developing countries, plants are the main medicinal source to treat various infectious
diseases. Plant extracts represent a continuous effort to find new compound against pathogens. Approximately 20% of
the plants are found in the world have been submitted to pharmacological or biological test, and a substantial number
of new antibiotics introduced on the market are obtained from natural or semisynthetic resources (Mothana and
Linclequist, 2005). Aloe vera (Aloe barbadensis miller) is a plant, which belongs to the family of Liliaceae and is
mostly succulent with a whorl of elongated, pointed leaves (Strickland et al., 2004; Beckford and Badrie, 2000). The
name is derived from the Arabic word ‘alloeh’ which means ‘bitter’, referring to the taste of the liquid contained in the
leaves. Aloe that is believed to have originated in the Sudan. Aloe vera grows in arid climates and is widely distributed
in Africa, India and other arid areas. The species is frequently cited as being used in herbal medicine. Aloe vera is a
perennial, drought resisting, succulent plant. It has stiff green, lance-shaped leaves containing clear gel in a central
mucilaginous pulp. Its thick leaves contain the water supply for the plant to survive long periods of drought (Foster,
1999). The leaves have a high capacity of retaining water also in very warm dry climates and it can survive very harsh
circumstances. When a leaf in cut, an orange-yellow sap drips from the open end. When the green skin of a leaf is
removed a clear mucilaginous substances appears that contains fibres, water and the ingredient to retain the water in
the leaf. The gel contains 99.3% of water, the remaining 0.7% is made up of solids with carbohydrates constituting for
a large components (Foster, 1999). Concentrated extracts of Aloe leaves are used as laxative and as a haemorrhoid
treatment. Aloe gel can help to stimulate the body’s immune system (Davis, 1997). The use of plant product for
pharmaceutical purpose has been gradually increased. According to World Health Organisation, medicinal plants
would be the best source for obtaining a variety of drugs (Santos et al., 1995). The use of plant extracts, with known
antimicrobial properties, can be of great significance in the treatment of various microbial infections. In the last
decade, numerous studies have been conducted in different countries to prove such efficiency in number of medicinal
plants. Most of the studies are restricted with crude extracts (Reddy et al., 2006; Erdo Urul, 2002; Atefl et al., 2003).

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Many scientific studies of the use of aloe vera have been undertaken, some of them conflicting. Despite these
limitations, there is some preliminary evidence that Aloe vera extracts may be useful in the treatment of wound and
burn healing, minor skin infections, Sebaceous cyst, diabetes, and elevated blood lipids in humans. These positive
effects are thought to be due to the presence of compounds such as polysaccharides, mannans, anthraquinones, and
lectins.
"The use of Aloes, the common musabbar, for external application to inflamed painful parts of the body and for
causing purgation [internal cleansing] are too well known in India to need any special mention."

MATERIALS AND METHODS

Collection of Plant Material:
The plant Aloe vera leaves were collected from in and around the city of Gulbarga, Karnataka, India.. This plant was
botanically authenticated in the Department of Botany, B.V. B college of UG and PG Bidar. The leaves were shade
dried and used for the extraction.
Extraction of Plant Material:
The leaves of Aloe vera was air dried and crushed to small piece using Mortar and Pestle and powdered in an electric
grinder. The powdered plant material was subjected for successive soxhlet extraction starting from non polar to polar
solvents such as petroleum ether [PE].Chloroform [CHCl3] and methanol [MeOH] by using soxhlet extracts. The
extracts were concentrated to dryness.
Preparation of the Extract:
The plant material were shade dried, powdered and subjected to Soxhlet extraction (1kg) with solvents ranging from
non-polar that is Petroleum ether (60-800), Chloroform (70-800), Methanol (60-900) respectively. The extracts were
concentrated to dryness in a flask evaporator under reduced pressure and controlled temperature. The petroleum ether
extract (20gm), chloroform (20gm), and ethanol extract (40gm). All the extracts were prepared in Tween-80 (1%)
suspended in distilled water. The extracts preparations were done as previously described by Alade and Irobi [6]. The
plant extracts were prepared by using soxhlet apparatus collected and stored in a vial for further studies.
Disc Preparation: The 6mm (diameter) discs were prepared from whatmann No. 1 filter Paper the discs were
sterilized by autoclave at 12°C. After the sterilization the moisture discs were dried on hot air oven at 50°C.Then
various solvent extract discs and control discs were prepared.
Antibacterial and Antifungal Activity Of Aloe Vera:
The antibacterial and antifungal activity studies were carried out by disc diffusion technique [12]. The sterile nutrient
agar plates and potato dextrose agar plates were prepared. The bacterial test organisms like Staphylococcus aureus,
Klebsiella pneumonia and Escherichia coli were spread over the nutrient agar plates by using separate sterile cotton
buds. Then the fungal test organism like Aspergillus niger and Candida were spread over the potato dextrose agar
plates After the microbial lawn preparation three different extracts of plant disc were placed on the organism
inoculated plates with equal significant difference between extract used and also distance control discs were also
prepared. All bacterial plates were incubated at 27°C for 24 hrs and fungal plates at 24°C for 72hrs. The diameter of
the minimum zone of inhibition was measured in mm. For each test, three replicates were performed.
Statistical Analysis: Data were expressed as mean±standard deviation. The data obtained were subjected to ANOVA
test to determine whether there was significant difference between extract used and also between the lengths of
incubation.

RESULTS
The present study carried out on the Aloe vera revealed to evaluate antimicrobial activities of various extracts of Aloe
vera.The successive leaf extracts using petroleum ether, chloroform and methanol of Alovera were tested for their
antimicrobial efficiency against pathogenic bacteria and fungi (Staphylococcus aureus, Klebsiella pneumonia, E.coli,)
and fungi like (Aspergillus Niger, Candida) at a dose 1: 20mg/ml and 2:40mg/ml. The standard drugs used for
comparison were Streptomycin and Fluconazole against bacteria and fungi. Among the extracted tested for their
antibacterial activity, the leaf extracts showed moderate to high activity against both gram positive and gram negative
bacteria. The extracts using petroleum ether, chloroform, and methanol of Aloe vera showed active antimicrobial
activity against Staphylococcus aureus, Klebsiella pneumonia, E.coli and, and antifungal activity against Candida and
Aspergillus niger.

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The chloroform and methanolic extract showed highest inhibition zone at higher concentration (i.e. 40mg/ml). Overall
the methanolic extracts showed greater inhibition of all pathogenic microorganisms used when compared to
chloroform and petroleum ether extracts.
The extracts of petroleum ether at the dose level of 20mg/ml showed the inhibition zone of Staphylococcus aureus
(15mm), Klebsiella (19mm), E.coli (14mm), (Fig. 1) whereas the extracts of petroleum ether at the dose level of
40mg/ml showed the inhibition zone of Staphylococcus aureus (20mm), Klebsiella (20mm), E-coli (10mm) (Plate-I,
& Fig. 2). The extracts of chloroform at dose level of 20mg/ml showed the inhibition zone of staphylococcus aureus
(15mm), Klebsiella (11mm), Escherichia coli (14mm) where as extracts of chloroform at the dose level of 40mg/ml
showed the diameter by zone of inhibition of Staphylococcus Aureus (14 mm), Klebsiella(10mm), E-coil (12mm)
(Table-1). The extract of methanol at the dose level of 20mg/ml showed the diameter of zone of inhibition of
Staphylococcus aureus (13mm), Klebsiella (13mm), E.coli (22mm) where as extracts of methanol at the dose level of
40mg/ml showed the diameter of zone of inhibition of Staphylococcus aureus (15mm), Klebsiella (15mm), E.coli
(16mm).

Table 1: Antimicrobial Activity Of Leaf Extract of Aloe vera

Zone Of Inhibition (mm)*
Leaf extracts Staphylococcus
Klebsiella sp. E. coil A. niger Candida
aureus
Chloroform (20mg) 15mm 11mm 14mm 10mm 15mm
Chloroform (40mg) 4mm 10mm 12mm 12mm 20mm
Petroleum ether (20mg) 15mm 19mm 14mm 12mm 14mm
Petroleum ether (40mg) 20mm 20mm 10mm 14mm 19mm
Methanol (20mg) 13mm 13mm 22mm 15mm 16mm
Methanol (40mg) 15mm 15mm 16mm 17mm 15mm
Standard (40mg) 12mm 20mm 30mm 15mm 28mm
Standard (20mg) 15mm 20mm 28mm 18mm 25mm
Data represents average of three replicates., mm* = Mean of three replicates.
Dose – 20 mg/ml and 40 mg/ml.

a b c

d e
a) Staphylococcus aureus b) Klebsiella pneumonia c) E. coli d) Candida e) Aspergillus niger.
Fig: I Plates Showing Zone of Inhibition

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32
30 28
28 25
26
24 22
22 19 20 Chloroform(20m
20 18 g)
18 1515 15 15 151416
16 1414 Petroleum(20mg)
14 13 13 12
12 11 10 Methanol(20mg)
10
8
6
4
2
0
S.Aureus Klebsiella E-coli A .niger Candida

Figure 1: Antimicrobial Activity of Leaf Extract of Aloe vera in 20mg Concentration

32 30
30
28
26
24
22 20 20 20 2019 Chloroform(40m
20
18 17 g)
15 15 16
16 14 15 15 Petroleum(40mg
14 12 12 12 )
12 10 10 Methanol(40mg)
10
8
6 4 4
4
2
0
S.Aureus Klebsiella E-coli A .niger Candida

Figure 2: Antimicrobial Activity of Leaf Extract of Aloe vera in 40mg Concentration

DISCUSSION
In this present study the petroleum ether extract, has shown high zone of inhibition in Escherichia coli, Klebseilla
pnemoniae, the fungi Aspergillus niger and moderate zone for Staphylococcus aureus, and Candida . Chloroform
extract has shown a high zone of inhibition in Klebsiella pneumonia, Escherichia coli and Aspergillus Niger but
moderate zone of inhibition in Staphylococcus aureus, and Candida. Methanol extract as shown high zone of
inhibition in Staphylococcus aureus, Klebseilla pnemoniae and Candida but moderate zone in Escherichia coli and
Aspergillus Niger. When compared the zone of inhibition with the standard drugs like streptomycin and flucanozole.
The plant extracts have shown almost equal to the standard drug. The above parameter supports the strong scientific
basis for the use of these plants in traditional treatment of microbial diseases.

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The antimicrobial activity of the extracts and their potency was quantitatively assessed by the presence or absence of
inhibition zone and zone diameter. Only alcoholic extract was found to be a better solvent for extraction of
antimicrobially active substances compared to water and hexane (Ahmad et al., 1998). Agarry et al., (2005) compared
the antimicrobial activities of the gel and leaf of Aloe vera against Staphylococcus aureus, Pseudomonas aeruginosa,
Trichophyton mentagraphytes, T. schoeleinii, Microsporium canis and Candida albicans. The antimicrobial analysis it
was confirmed that this plant leaf extracts showed positive results against bacterial species such as Klebsiella
pneumoniae, Staphylococcus aureus and Escherichia coli and fungi Aspergillus niger and candida. Hence, it can be
concluded that the leaf extracts of Aloe Vera can effectively act as an antimicrobial agent which have ability to replace
most of medium medicines of this era.

CONCLUSION
The present study has revealed the importance of natural products to control antibiotic resistant bacteria, which have
been a threat to human health. It is, therefore highly essential that medicinal plants whose properties have not been
fully characterized should form a top agenda of top management in developing nations whose citizens are sometimes
unable to afford expensive orthodox medicine. This study has revealed the presence of many secondary metabolites in
the leaves of Aloe vera. It has the further confirmed that the plant extracts could be used for the treatment of various
infections including skin transmitted infections. The results lend credence to the folkloric use, if this plant in treating
microbial infection and shows that Aloe vera could be exploited for new potent antimicrobial agents.

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