Investigation on antimicrobial and antioxidant

properties of Samanea saman

Thippeswamy, S., Mohana, D.C*and Abhishek, R.U.
Department of Microbiology and Biotechnology, Bangalore University, Jnanabharathi Campus,
Bangalore - 560 056, Karnataka, India. *E-mail: mohanadc@gmail.com
ABSTRACT
The present investigation evaluates the antimicrobial and antioxidant
activities of successive solvent extracts and isolated constituents of Samanea
saman. The methanol extract and alkaloid constituent exhibited the highest
antibacterial activity against Gram positive and Gram negative bacteria, yeast
and moulds, followed by ethanol extract. Among bacterial strains tested, S.
faecalis was most sensitive, while P. vulgaris was highly resistant. The human
pathogenic fungi viz., Candida albicans and Cryptococcus neoformans were
strongly inhibited at 1mg/ml. Among the plant pathogenic fungi tested, Fusarium
moniliforme was highly sensitive, while the most resistant was Penicillium
citrinum.
The antioxidant activity was assessed by DDPH radical scavenging assay.
The methanol extract and alkaloid constituent showed significant DPPH radical
scavenging activity at 1mg/ml (90.78%), whereas synthetic antioxidant BHT
showed 92.64% at 1 mg/ml. The present investigation is an important step in
developing plant based antimicrobial and antioxidant drugs, which are ecofriendly for the management of the diseases caused by pathogenic microbes and
free radicals. Further investigations are necessary for developing commercial
formulation based on animal trails and toxicological experiment.

INTRODUCTION
The number of multi drug resistant microbial strains and the appearance of
the strains with reduced susceptibility to antibiotics are continuously increasing
(Dabur et al., 2007). The increasing failure of chemotherapeutics and antibiotic
resistance exhibited by pathogenic bacteria, need to look alternative strategies
for the management of antibiotic resistant microbes. Thus there is urgent need to
look for alternative strategies for the management of drug resistant pathogenic
microbes. Further, free radicals are the causative agents of protein oxidation,
DNA damage and lipid peroxidation in living cells (Kil et al., 2009). Oxidation of
lipids causes undesirable effects in human beings, it is essential to decrease
lipid peroxidation products in foods. The use of synthetic antioxidnats in food
processing has led to the appearance of remarkable side effects (Ebrahimabadi
et al., 2010). In recent decades, interest considerably increased for finding
naturally occurring antioxidant and antimicrobial compounds suitable for use in
food or medicine.
The plant derived substances have recently become of great interest owing to
their versatile applications in medical and agriculture system (Sharma et al.,
2009). This is also true in India and only a small percentage of plants of this
region have been evaluated for antimicrobial activity against plant and human
pathogenic microbes. Some compounds may have antimicrobial and antioxidant
activities associated with the reported health effects (Bystrom et al., 2009). This
led the authors to screen in vitro, a large number of plants for antimicrobial and
antioxidant activities, with the ultimate aim of developing plant based formulation
for plant and human disease management .
Samanea saman (Jacq.) Merr. is belongs to the family Fabaceae and it is globally
distributated throughout the tropical regions. It is used as folk remedy for curing
various diseases (Prasad et al., 2008). A scientific and systematic investigation
with regards to the various biological activities of this plant is lacking. Thus
considering vast potentiality of plants as a source of new chemotherapeutic
agents, detailed investigations was conducted to test the efficacies of S. saman
against some pathogenic bacteria and fungi, and its antioxidant efficacies.

MATERIALS AND METHODS

The tested pathogenic bacteria and yeast were collected from National
collection of Industrial Microorganisms (NCIM), Pune and seed borne
phytopathogenic moulds were isolated from maize, paddy and sorghum seeds by
standard blotter method and agar plate method. All analytical grade chemicals,
culture media, reagent and solvents were obtained from Hi media and Sisco
Research Laboratory, Mumbai.
The plant material was collected from southern part of Karnataka. The plant
material was shade dried, powdered and subjected to successive solvent
extraction in soxhlet apparatus. All the solvent extracts were tested for their
antimicrobial and antioxidant properties. The active extract was further subjected
to active component separation by fractionation and preparative TLC method.
Disc diffusion method was employed for assessing antimicrobial activity
against bacteria and yeasts. Briefly, 5mm filter paper discs were impregnated
with desired different concentrations of extract and isolated constituents and
placed on pre-inoculated (108 CFU of bacteria and 106 of yeast) media plates and
incubate the plates for specific time and temperature. After incubation
measured the zone of inhibition around the disc. Whereas, poison food technique
was employed for moulds, in this method desired different concentrations of
extract and isolated constituents were incorporated in to the media and the 5 mm
disc of 7 day old culture of the test fungi were inoculated. The petridish
containing media devoid of extract served as control. The plates were incubated
at 26
10C for seven days. The Percentage Inhibition (%) of mycelial growth was
calculated by using the formula,
Percentage Inhibition = dc-dt X 100/dc
Where, dc - Average increase in mycelial growth in control. dt - Average increase in
mycelial growth in treatment

DPPH radical scavenging assay was used to determine the antioxidant

activities of solvent extracts and isolated constituents. Briefly 1 ml of different
concentrations of solvent extracts, isolated constituents and butylatedhydroxy
toluene were mixed with 3 ml of 0.1Mm DPPH solution separately. All the tubes
were incubated in dark for 30 min. After incubation measure the reduction in
purple colour by spectrophotometer at 517nm and caluculated the DPPH radical
scavenging activity by using following formula
I % = {(AcontrolAsample)/ Acontrol} X 100
Where Acontrol is the absorbance of control, Asample is the absorption of the extracts.

RESULTS
Antibacterial and anticandidal activity of S. saman is presented in table-1.
Among the bacteria and yeasts tested S. faecalis was the most sensitive and P.
vulgaris was most resistant. The antifungal activity result is presented in Table-2.
Antioxidant activity of different solvent extract of S. saman is presented in Graph1. Among the extracts tested methanol showed highest antioxidant activity
presented in graph-2.

Table-1: Antibacterial and anticandidal activity of S. saman and synthetic

antibiotic/ antifungal compounds against some human and plant
pathogenic bacteria and yeast

Organisms

Aqueous
extract

E. coli
K. pneumoniae
P. vulgaris
P. aeroginosa
S. typhimurium
S. aureus
S. faecalis
X. campestris
C. albicans
C. neoformans

7.30.3
9.00.3
00.0
00.0
7.30.3
12.60.3
25.60.3
9.10.1
12.80.4
15.30.6

Solvent extracts
(1 mg/ml)
Methanol

Ethanol

Synthetic antibiotic/
antifungal compounds
Augmentin Amphotericin-B
(30mcg)
(100 mcg)

Zone of Inhibition (mm)

12.90.5
13.00.4
12.20.4
15.70.4
11.01.1
14.70.5
30.50.5
150.4
15.60.6
22.30.8

120.7
11.00.4
8.50.2
100.5
9.71.7
13.50.5
26.50.2
11.00.4
10.00.6
17.00.5

8.00.4
6.00.3
6.00.3
28.00.6
16.00.5
20.00.5
23.00.7
0.0
-

10.00.2
17.00.4

Table-2: Antifungal activity of aqueous, different solvent extracts and

isolated constituents of leaves of S. saman against some
phytopathogenic fungi
Test organisms

Aqueous extract

Solvent extracts
IC50 value
(1 mg/ml)
(mg/ml)
Methanol
Ethanol
74.70.2 45.10.4
0.5
64.20.4 50.00.5
1.0
21.30.5 22.50.6
3.0
36.80.5 24.30.5
3.0
15.00.4
7.70.5
4.0
73.00.6 53.20.7
0.5
66.902
39.10.4
0.5
71.90.2 66.93.1
0.5
81.60.4 43.80.6
0.3
43.40.2 28.41.2
0.5
70.20.8 58.80.9
0.5

Synthetic
fungicide
DM-45
79.10.2
68.90.3
47.60.3
63.10.1
24.40.3
78.90.3
74.00.6
58.90.3
69.80.8
69.10.1
68.30.3

Al. brassicicola
Al. geophila
A.flavus
A. fumigatus
A. tamari
C. tetramera
F. equiseti
F. lateratium
F. moniliforme
F. oxysporum
F. udum

10%
50.00.3
5.21.2
6.61.5
22.10.8
15.51.1
53.10.3
61.80.3
46.40.0
63.50.6
8.60.2
51.31.2

50%
72.20.6
91.60.0
42.15.5
46.00.5
62.90.6
77.10.8
89.00.3
87.20.3
91.00.0
35.20.4
90.50.0

P. chrysogenum

20.40.3

52.60.3

180.6

140.4

5.0

72.40.3

P. citrinum

27.90.3

77.90.0

15.50.5

7.80.5

5.0

63.90.3

Graph-1. DPPH radical scavenging

activity of solvent extracts at
1mg/ml.

Graph-2.
DPPH radical scavenging
activity of methanol extract at different
concentrations (0.031-1 mg/ml).

CONCLUSION
In recent decades natural antimicrobials, antioxidants and chemotherapeutics
gaining importance due to sever side effects of synthetic drugs. The present
investigation is an important step in developing plant based antimicrobial and
antioxidant drugs, which are eco-friendly for the management of the disease
caused by pathogenic microbes and free radicals.

ACKNOWLEDGEMENTS
The authors are grateful to thank University Grant Commission (UGC), New
Delhi and Department of Science and Technology (DST), New Delhi for providing
financial support.