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Appendix
Antibacterial Activities of Green Algae (Chlorophyceae) From Narmada River
Shailendra Sharma, Taniya Sengupta
Published as a Chapter in Floral diversity and their conservation, D. K Agencies (P) LTD
New Delhi. ISBN- 987-81-7622-236-0
ABSTRACT
This paper deals with the objective to isolate green algae ―Chlorophyceae‖ from Narmada
River and to study its antimicrobial activity. The isolated algae were of two different types
one was the fresh algae and the other was the dried form collected from the shore line area of
Narmada River. Ethanol, chloroform, water, glycerol, acetone, formaldehyde, phenol and
acetic acid crude extract from algae were obtained and tested against one Gram‘s Positive
bacteria ―Bacillus subtilis‖ and another against Gram‘s Negative bacteria ―Escherichia coli‖.
The maximum antibacterial activity was recorded from formaldehyde extracts of both types
of algae against both bacteria. No antibacterial activity was recorded from water extract. The
minimum activity was recorded from dried algae extract against Bacillus subtilis as compared
to fresh one whereas the dried extract was pointed out to give maximum inhibition against
Escherichia coli as compared with the fresh one. The present finding indicates the need for
detailed work on the aspect, especially with reference to antimicrobial activities against
different pathogenic bacteria.
KEY WORDS: Chlorophyceae, Escherichia coli, Bacillus subtilis, Antibacterial activities,
Narmada River.
Correspondence Author
Taniya Sengupta
INTRODUCTION
The Narmada is a river in central India and
the fifth largest river in the Indian
subcontinent. It forms the traditional
boundary between North India and South
India and flows westwards over a length of
1,312 km (815.2 mi) before draining
through the Gulf of Cambey (Khambat)
into the Arabian Sea, 30 km (18.6 mi)
west of Bharuch city of Gujarat.[1] It is
one of only three major rivers in
peninsular India that runs from east to
west (largest west flowing river) along
with the Tapti River and the Mahi River. It
flows through the states of Madhya
Impact of sociobiological activities on Narmada River flowing from Omkareshwar to Khalghat (M.P.)
213
Pradesh (1,077 km (669.2 mi)),
Maharashtra, (74 km (46.0 mi)) – (35 km
(21.7 mi)) border between Madhya
Pradesh and Maharashtra and (39 km (24.2
mi) border between Madhya Pradesh and
Gujarat and in Gujarat (161 km (100.0
mi)). The Narmada basin, hemmed
between Vindya and Satpuda ranges,
extends over an area of 98,796 km2
(38,145.3 sq mi) and lies between east
longitudes 72 degrees 32' to 81 degrees 45'
and north latitudes 21 degrees 20' to 23
degrees 45' lying on the northern extremity
of the Deccan Plateau. The basin covers
large areas in the states of Madhya
Pradesh (86%), Gujarat (14%) and a
comparatively smaller area (2%) in
Maharashtra. In the river course of
1,312 km (815.2 mi) explained above,
there are 41 triburaries, out of which 22
are from the Satpuda range and the rest on
the right bank are from the Vindhya range.
Algae are large and diverse group of
simple typically autotrophic organisms,
ranging from unicellular to multi cellular
forms. They have their pigments localised
in membrane bounded intracellular bodies
(Plastids) have no vascular system and do
not develop from an embryo. Algae are an
extremely important species they can
produce more oxygen then all the plant in
the world. They form an important food
source for many animals. Thus they are at
the bottom of the food chain with many
living thing depending upon them.
Chlorophyceae have primary chloroplasts,
i.e. the chloroplasts are surrounded by two
membranes and probably developed
through a single endosymbiotic event.
Green Algae have chloroplasts with
chlorophyll a and b.Higher plants are
pigmented similarly to Green Algae and
probably developed from them.
To date, there is moderately an assortment
of reports commerce with the antibacterial
activity of solvent extract attained from
freshwater algae. Antimicrobial activities
in freshwater algae have been reported
wide reaching for decades by various
scientist (Debro et al;1979, Barchi et al;
1984, De Mule et al; 1991, Ishida et al;
1997, Ozdemir et al;2001, Noaman et
al;2004). An effort has been derelict in the
present study to explore the antibacterial
activities of freshwater algae of Narmada
River on both Grams positive and Grams
negative bacteria.
MATERIALS AND METHODS
The green algae was collected from the
shore line areas of Narmada River from
Mandleshwar in two forms one is the fresh
and the other war the dried form. The
plants were washed in tap water and
cleaned of all epiphytes and other attached
material and salt as soon as possible after
collection. Then the plants were air dried
for few days in the shade.
2 grams of both fresh and dry algae were
soaked into 10ml of each solvent (Acetic
acid, acetone, chloroform, ethanol,
formaldehyde, glycerol, phenol and water)
for five days. The resultant crude extracts
were filtered. The crude extracts were kept
in 4 until testing their antibacterial
activities against Gram‘s negative Bacteria
Escherichia coli and Gram‘s negative
Bacteria Bacillus subtilis were done. For
testing the antibacterial activities Plates of
Muller Hinton (Himedia) have been used.
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214
RESULTS AND DISCUSSIONS
Table 01: Antibacterial activity of fresh green algae Chlorophyceae obtained from Narmada
Shore line areas.
Sr.
no Extract prepared in
Zone of Inhibition (cm)
Escherichia coli Bacillus subtilis
1 Acetone 00 0.8
2 Acetic Acid 2.8 3.0
3 Chloroform 2.0 00
4 Formaldehyde 6.5 6.0
5 Glycerol 00 00
6 Phenol 4.2 3.0
7 Water 00 00
Table 02: Antibacterial activity of Dried green algae Chlorophyceae obtained from Narmada
Shore line areas.
Sr.
no Extract prepared in
Zone of Inhibition (cm)
Escherichia coli Bacillus subtilis
1 Acetone 1.2 00
2 Acetic Acid 2.3 2.6
3 Chloroform 1.5 0.8
4 Formaldehyde 7.2 7.8
5 Glycerol 3.6 3.4
6 Phenol 3.8 00
7 Water 00 00
On completion of 24 hrs incubation the
zone of inhibition were measured in each
plate for both the kind of green algae.
Table 01shows the inhibition zone for
each extract prepared from fresh green
algae. Algae itself in water does not
release any extracellular components for
inhibiting the growth of bacteria and in
organic solvents except acetone and
glycerol the fresh green algae shows the
activities against Escherichia coli. Against
Bacillus subtilis the fresh algae also shows
remarkable inhibition except chloroform
and Glycerol as a solvent. Maximum
inhibition had seen in both bacteria from
formaldehyde extract that is for
Escherichia coli and Bacillus subtilis 6.5
and 6.0 respectively (Fig 01).
Table 02 shows the results for the dried
form of green algae against both selected
bacterial species. Alike fresh form the
dried form of the green algae also did not
show any anti bacterial activities. The
other organic extracts show efficient
inhibition against Escherichia coli,
whereas in case of Bacillus subtilis the
results were positive except acetone and
phenol extracts. Here also the most
effective results were given by
formaldehyde extracts against Escherichia
coli and Bacillus subtilis as 7.2 and 7.8
respectively (Fig 02).
\
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215
Formaldehyde Fresh Green Algae
Escherichia coli Bacillus subtilis
Formaldehyde Dried Green Algae
Escherichia coli Bacillus subtilis
On comparing it had been concluded that
Formaldehyde extracts has been proved to
be most acceptable antibacterial agent
against pathogenic bacteria. Another thing
which have been concluded was that the
fresh algae extract was much effective on
Gram‘s negative bacteria (Escherichia
coli) and the dried form shows more
response against Gram‘s positive bacteria
(Bacillus subtilis).
Further work have to be done to determine
the effect of different solvents used for
extract preparation on the bacterial growth
and also other different types of bacteria
have to be chosen to find how and to what
Fig 01: Zone of inhibition against both types of bacteria by Formaldehyde
fresh green algae extract.
Fig 02: Zone of inhibition against both types of bacteria by Formaldehyde
dried green algae extract.
Impact of sociobiological activities on Narmada River flowing from Omkareshwar to Khalghat (M.P.)
216
extent these extracts can be used as
antibacterial agent which could control the
growth of lethal pathogenic bacteria.
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