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Research Article
Article History
Received : 22.04.2015
Revised : 29.04.2015
Accepted : 08.05.2015
1. Introduction
D. Sekar / Life Science Archives (LSA), Volume 1, Issue 4, Page: 250 - 256, 2015
activities (Tuney et al., 2006; Patra et al., 2008).
Many bioactive compounds of marine macroalgae
with pharmacological activity have been isolated
and some of them are still under investigation to
protect the various infections including life style
related diseases. Some of these compounds are
sterols, terpenoids, polysaccharides, peptides,
proteins, vitamins, acrylic acid, terpenes,
chlorophyllides, phenols, heterocyclic compounds,
halo-genated ketones and alkanes and cyclic
polysulphides (Priyadharshini et al., 2011).
251
and
inoculum
The test bacterial isolates were subcultures and maintained on Nutrient agar slants
and stored in refrigerator at 4C.
Bacterial inoculums preparation
Bacterial inoculums was prepared by
inoculating a loopful of test organisms in 5 ml of
Nutrient broth and incubated at 37 C for 3 - 5
hours till a moderate turbidity was developed. The
turbidity was matched with 0.5 McFarland
standards and then used for the determination of
antibacterial activity.
Disc preparation
D. Sekar / Life Science Archives (LSA), Volume 1, Issue 4, Page: 250 - 256, 2015
method proposed by Bauer et al. (1966). Petri
plates were prepared by pouring 20 ml Muller
Hinton agar and allowed to solidify for the use in
susceptibility test against bacteria. Plates were
dried and 0.1 ml of standardized inoculums
suspension was poured and uniformly spreaded.
The excess inoculums were drained and the plates
were allowed to dry for five minutes. After drying,
the discs with extract were placed on the surface
of the plate with sterile forceps and gently pressed
to ensure contact with the agar surface. The
Ampicilin (5 g/disc) was used as positive control
and the 5 % DMSO was used as a blind control in
these assays. The plates were incubated at 37 C
for 24 hours. The zone of inhibition was observed
and measured in millimeters. Each assay in these
experiments was repeated three times for
concordance.
Minimum inhibitory concentration for bacteria
Minimum inhibitory concentration (MIC)
of the seaweed extracts against bacterial isolates
was tested in Muller Hinton broth by Broth macro
dilution method (Ericsson and Sherri, 1971). The
seaweed extracts were dissolved in 5 % DMSO to
obtain 128 mg/ml stock solutions. A quantity of
0.5 ml of stock solution was incorporated into 0.5
ml of Mueller Hinton broth for bacteria to get a
concentration of 64, 32, 16, 8, 4, 2 and 1 mg/ml
for seaweeds extracts and 50 l of standardized
suspension of the test organism was transferred on
to each tube. The control tube contained only
organisms and devoid of seaweed extracts. The
culture tubes were incubated at 37C for 24 hours.
The lowest concentration, which did not show any
growth of tested organism after macroscopic
evaluation was determined as Minimum inhibitory
concentration (MIC).
3. Results and discussion
Antibacterial
activity
of
marine
macroalgae against Staphylococcus aureus
The antibacterial activity of selected
marine macroalgae crude extracts was determined
against pathogenic Gram positive bacteria
Staphylococcus aureus and the results were
252
D. Sekar / Life Science Archives (LSA), Volume 1, Issue 4, Page: 250 - 256, 2015
253
Table - 1: Antibacterial activity of marine macro algae crude extract against Staphylococcus aureus
Concentration of the marine macroalgae crude extract (mg/ml) and Zone of inhibition (mm)
Name of the marine
macroalgae
Turbinaria conoides
Sargassum wightii
Padina gymnospora
Caulerpa racemosa
Acanthophora spicifera
Hexane
Chloroform
Ethyl acetate
Acetone
Positive
control*
Methanol
150
mg/ml
100.8
100.7
110.7
90.7
300
mg/ml
110.6
110.4
130.7
100.6
150
mg/ml
100.6
110.6
140.4
110.5
300
mg/ml
130.5
120.7
150.5
120.4
150
mg/ml
130.4
120.5
140.7
120.2
300
mg/ml
150.7
140.4
160.6
130.6
150
mg/ml
140.4
130.3
150.4
130.6
300
mg/ml
170.4
160.5
180.6
140.3
150
mg/ml
180.6
160.6
190.6
150.6
300
mg/ml
190.4
170.3
200.0
160.5
90.4
100.4
110.4
120.2
110.6
130.0
130.3
140.0
130.5
160.0
20
20.6
Hexane
Chloroform
Ethyl acetate
Acetone
Methanol
Positive Control*
320
320
160
320
640
160
160
80
160
320
80
80
40
80
160
40
40
20
40
80
20
20
20
20
40
20
20
20
20
20
Turbinaria conoides
Sargassum wightii
Padina gymnospora
Caulerpa racemosa
Acanthophora spicifera
*Ampicillin (20 g)
Table - 3: Antibacterial activity of marine macro algae crude extracts against Streptococcus pyogenes
Concentration of the marine macro algae crude extracts (mg/ml) and Zone of inhibition (mm)
Name of the
marine
macroalgae
Hexane
Chloroform
Ethyl acetate
Acetone
Positive
control*
Methanol
150
mg/ml
300
mg/ml
150
mg/ml
300
mg/ml
150
mg/ml
300
mg/ml
150
mg/ml
300
mg/ml
150
mg/ml
300
mg/ml
100.7
120.8
120.6
130.6
130.3
150.6
140.4
160.9
170.5
190.7
100.4
110.7
110.6
130.3
120.5
140.4
130.6
150.7
160.2
180.4
110.4
130.6
130.0
140.4
140.3
160.6
150.6
170.6
180.4
210.5
90.6
110.3
Acanthophora
90.2
100.6
spicifera
Mean SD, *Ampicillin (20 g)
110.3
120.6
120.4
130.7
130.5
150.3
140.5
160.4
100.3
110.7
120.3
130.4
120.8
140.3
130.6
150.3
Turbinaria
conoides
Sargassum wightii
Padina
gymnospora
Caulerpa racemosa
20 g
220.4
Table - 4: Minimum inhibitory concentration of marine macro algae crude extracts against Streptococcus pyogenes
Minimum inhibitory concentration of marine macro algae crude extracts (mg/ml)
Name of the marine
macroalgae
Turbinaria conoides
Sargassum wightii
Padina gymnospora
Caulerpa racemosa
Acanthophora spicifera
*Ampicillin (20 g)
Hexane
Chloroform
Ethyl acetate
Acetone
Methanol
Positive Control*
320
320
160
640
640
160
160
160
320
320
80
80
80
160
160
80
80
40
80
80
40
40
20
40
40
20
20
20
20
20
D. Sekar / Life Science Archives (LSA), Volume 1, Issue 4, Page: 250 - 256, 2015
254
Table - 5: Antibacterial activity of marine macro algae crude extracts against Bacillus cereus
Concentration of the marine macro algae crude extracts (mg/ml) and Zone of inhibition (mm)
Name of the marine
macroalgae
Hexane
150
mg/ml
110.6
100.7
120.6
90.6
Turbinaria conoides
Sargassum wightii
Padina gymnospora
Caulerpa racemosa
Acanthophora
90.2
spicifera
Mean SD, *Ampicillin (20 g)
Chloroform
Ethyl acetate
Acetone
Positive
control*
Methanol
300
mg/ml
130.4
120.4
150.4
110.7
150
mg/ml
130.7
110.7
130.7
110.9
300
mg/ml
140.5
130.6
150.8
130.3
150
mg/ml
140.2
130.4
140.7
110.7
300
mg/ml
160.6
150.5
170.6
140.4
150
mg/ml
140.8
140.4
160.4
130.3
300
mg/ml
160.9
150.8
180.9
150.4
150
mg/ml
160.3
140.6
170.5
140.2
300
mg/ml
180.3
170.6
190.4
160.4
110.3
100.9
120.6
110.2
130.6
120.7
140.5
130.5
150.5
20 g
210.6
Table - 6: Minimum inhibitory concentration of marine macro algae crude extracts against Bacillus cereus
Minimum inhibitory concentration of marine macro algae crude extracts (mg/ml)
Name of the marine
macroalgae
Turbinaria conoides
Sargassum wightii
Padina gymnospora
Caulerpa racemosa
Acanthophora spicifera
*Ampicillin (20 g)
Hexane
Chloroform
Ethyl acetate
Acetone
Methanol
160
160
160
320
640
80
160
80
160
320
80
80
40
160
320
20
40
20
80
80
20
20
20
40
40
Positive
Control*
20
20
20
20
20
D. Sekar / Life Science Archives (LSA), Volume 1, Issue 4, Page: 250 - 256, 2015
followed by Turbinaria conoides (18 0.3 mm),
Sargassum wightii (17 0.6 mm), Caulerpa
racemosa (16 0.4 mm) and Acanthophora
spicifera (15 0.5 mm). The crude hexane extract
of marine macroalgae showed minimum zone of
inhibition against Bacillus cereus when compared
to the other solvent extracts. No zone of inhibition
was observed in DMSO negative control and the
positive control Ampicillin (20 g) showed 21
0.6 mm zone of inhibition against the Bacillus
cereus. The minimum inhibitory concentration
(MIC) of marine macroalgae crude extracts
against Bacillus cereus was ranged between 20
mg/ml to 640 mg/ml and the results were showed
in Table - 6. The crude methanolic extract of
Padina gymnospora, Turbinaria conoides and
Sargassum wightii showed best MIC at 20 mg/ml
against Bacillus cereus. The extract of
Acanthophora spicifera and Caulerpa racemosa
recorded best MIC at 40 mg/ml.
The results of the present research showed
that seaweed extract of Padina gymnospora
showed maximum inhibitory activity against
Gram positive bacteria and the findings obtained
from the study of Salvador et al. (2007) was well
matched with the present study.
In the study carried by Salvador et al.
(2007), the extracts of Padina gymnospora were
prepared and screened for its antibacterial activity.
Padina gymnospora extract that was prepared and
it showed highest inhibitor activity against Gram
positive bacteria when compared to Gram negative
bacteria. The results of the present research
showed that seaweed extract of Padina
gymnospora showed maximum inhibitory activity
against Gram positive bacteria and the findings
obtained from the study of Salvador et al. (2007)
was well matched with the present study.
4. References
1. Baker, J.T. 2014. Seaweeds in Pharmaceutical
studies and applications. Hydrobiology, 116
(117): 29-40.
2. Bauer, A. W., W. M. M. Kirby, J. C. Sherris
and M. Turck. 1966. Antibiotic susceptibility
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