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Life Science Archives (LSA)

ISSN: 2454-1354
Volume 1; Issue - 2; Year 2015; Page: 112 - 123

Research Article
In vitro STUDIES OF ANTIFUNGAL AND ENZYMATIC ACTIVITY OF
SELECTED FUNGI ISOLATED FROM WATER USING DIFFERENT BAITS
S.A. Aghizion Inbakani 1, V. Bhuvaneswari*2, G. Kathiravan3 and B. Shanmugapriya2
1
S.D.N.B. Vaishnav College for Women, Chromepet, Chennai - 600 044, Tamil Nadu, India.
2
Chikkaiah Naicker College, Veerapan chitram, Erode 600 004, Tamil Nadu, India.
3
Department of Biotechnology ,Vels University, Pallavaram, Chennai 600 117, Tamil Nadu, India.
Abstract
Aquatic fungi present in the fresh water ecosystem are of biological importance. Some serve as a
food for freshwater crustacean and other organisms. Some are parasitic on freshwater fishes while many are
destructive in aquarium and fish hatcheries. In this study, enumerations of fungi were carried out using moist
chamber incubation and baiting technique. Seeds, plants and animal segments, insects, fruits and vegetables
were used as baits. A total of 4614 fungal colonies were isolated. The relative percentage of the individual
groups of fungi revealed that hyphomycetes was maximum followed by zygomycetes, sachharomycetes,
ascomycetes, sterile morphospecies, coelomycetes and oomycetes. In addition to that the isolated fungi were
tested for enzyme activities such as amylase, laccase, and lipolytic activity. In vitro antifungal activity of
methanol extracts of some Indian medicinal plants against test fungi was done by agar disc diffusion method
to evaluate its potential importance. All the experimental test fungi subjected to enzyme assay showed
positive results for amylase activity whereas laccase activity was observed in only Trichoderma viride. Out
of the methanol extracts of the five medicinal plants tested, Boerhavia diffusa, Lantana camara, and Ricinus
communis showed best antifungal activity against Aspergillus flavus, Cladosporium cladosporioides, and
Drechslera halodes. Thus, this work shows that the aquatic environment is blessed with abundant supply of
microorganisms waiting to be explored in various areas such as biodegradation, waste management etc.
Key words Aquatic fungi, Enzyme activity,
Medicinal plants and Plant extract.

Article History
Received : 21.03.2015
Revised : 01.04.2015
Accepted : 06.04.2015
1. Introduction

Fungi are universally present in all types

of natural waters and form one of the most
important components of an ecosystem as
decomposers. Mycelia frequently appear on seeds,
fruits, petals, leaves, twigs and other elements of
plants fallen into water (Kiziewic, 2005). Baiting
techniques have provided a wealth of information
on isolation & distribution of aquatic fungi.
* Corresponding author: V. Bhuvaneswari
Tel.: +91-9176599550
E-mail: bhuvneshwari.v@gmail.com

Several baits autoclaved wood cubes, filter paper

strips, dead fishes, meat piece, dead flies, insect
larvae, cooked egg white, and discs of cork have
been used to isolate aquatic fungi (Alabi, 1971;
Sharp 1978; Agina and Kpu, 1988). The use of
hemp seeds as baits (Sparrow, 1960; Lui and Volz,
1977; Sharp, 1978) for aquatic fungi is popular.
Zoosporic fungi from different water bodies have
been studied in many parts of the world by
numerous researchers (Ziegler, 1958; Roberts,
1963; Alabi, 1971 a & b, 1974; El-Hissy, 1994).
Some fruits such as lemon, oranges, apples &

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

pears are very good as baits for aquatic

phytophthora species (Adenle, 1994). The aim of
this study is to access the suitability of different
kinds of baits for the enumeration of fungi
associated with water and the effect of the
ecological factors including certain physical &
chemical parameters of water & composition of
the substratum from which the fungi were isolated.
In addition to that the isolated fungi were tested
for enzyme activities such as amylase, laccase,
and lipolytic activity. In vitro antifungal activity
of methanol extracts of some Indian medicinal
plants against test fungi was done by agar disc
diffusion method to evaluate its potential
importance.
2. Methodology
General microbiological and laboratory
techniques followed in the present investigation
were as there outlined by Booth (1971). In the
present investigation, four kinds of water samples
like pond, canal, lake and tap water were collected
from ten different locations. Water samples
employed in the study was collected in and around
Chennai, early in the morning usually between
5.00 a.m. and 7.00 a.m. and stored in polythene
bags. About 1000 ml water samples were collected
and used for further experimentation. The list of
location and kind of water samples are presented
in Table - 1.
Attempts were made to study and analyze
the physicochemical characteristics of the water
(El-Hissy et al, 1990). The following parameters
were analyzed in Tamil Nadu Water Supply and
Drainage (TWAD) Board, State Level Water
Testing
Laboratory,
Chennai
600 005. Physical examinations - Colour, odour,
turbidity, electrical conductivity, total dissolved
solids & suspended solids. Chemical examinations
- Alkalinity, pH, hardness, sodium, potassium,
iron, manganese, ammonia, nitrate, nitrite,
chloride, fluoride, sulphate, phosphate, calcium
carbonate, and silica.
2.1. Methodology for recovery
associated with water samples
the

of

fungi

Isolation of fungi was carried out by using

following isolation techniques
viz.,

113

(a) Incubation and (b) baiting techniques in the

laboratory.
2.1.1. Incubation Method
In incubation method, small fragments of
substrates of plant decaying leaf litter, aquatic
plant parts, woody materials and animal origin
(fish scales, gills, fins and fish tails) were
collected from the lake. The materials were broken
into small pieces and incubated on wet blotters in
petriplates. The materials along with petriplates
were kept in the incubator under laboratory
condition (222C temperature) for about 8 days
(Czeczuga, 1991 a & b). At the end of the
incubation period, the colonized fungi on
incubated
materials
were
examined
microscopically and the fungal hyphae were
transferred to a sterilized petriplate containing
culture media. The following culture media
namely, Water Agar Medium (WAM), Potato
Dextrose Agar (PDA) Medium, Corn Meal Agar
(CMA) Medium, Glucose Yeast (GY) Medium
and Glucose Yeast Peptone (GYP) Medium was
used for the enumeration of fungi.
2.1.2. Baiting Technique
Seeds were used as baits to isolate
zoosporic fungi (Farida et al., 2001; Kiziewicz,
2005). The selected seeds were placed in one litre
containers and covered properly to protect the
water from penetration by bacteria. After a few
days of incubation, microscopically determined
mycelia were removed from the seeds and
transferred to culture media amended with
chloramphenicol and cycloheximide (150 mg/L)
to inhibit bacterial growth (Roberts, 1963). These
petridishes were examined periodically for
identification.
For the isolation of zoosporic fungi
associated with aquatic plant and animal
segments, samples were collected from the same
location and washed thoroughly under tap water to
remove the debris and surface sterilized with 1%
mercuric chloride for 10 min and rinsed with
distilled water for about 5 min and then finally
dried on a sterilized filter paper. The surface
sterilized segments were platted on culture media
for the enumeration of fungi.

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

2.2. Methodology for recovery

associated with mud samples

of

fungi

The following baiting technique was used

for the recovery of fungi from mud samples (ElHissy and Abal-Eloah, 1989). A 50 g of each mud
sample was introduced into clean sterile
petridishes. The mud sample in each petridish was
then flooded with sterile distilled water with
addition of few sesame seeds. The submerged mud
samples were also processed using certain
vegetables as bait. In this method, the selected
vegetable baits were swabbed with alcohol and a
hole of approximately 10 mm diameter was cut
through to the core on one side using a sterile cork
borer. The hole was packed with soil and covered
with sticky tape to retain the soil. The baits were
incubated at room temperature in the light for 4 to
5 days. Isolations were made after the emergence
of mycelium from the baits.
2.3. Incubation, isolation and identification of
fungi
The petriplates were incubated in a light
chamber and observations were done from the
second day onwards for a period of 3 - 4 weeks for
the fungal colonies (Kiziewicz, 2005). The light
regime was 12 hours light followed by 12 hours
darkness. The hyphae, which grew out from the
bait samples were transferred to fresh PDA slants.
They were maintained by sub-culturing. To
prevent the rapidly growing fungi from inhibiting
the slow growing species, the former were
removed as soon as they appeared on the plates.
The identification of fungi was based on
morphological features such as shape and size of
hyphae, shape of sporangium and spores, structure
of oogonium, oosporum, and antheridium.
Identification and characterization of fungi were
made as per the key and standard monographs.
2.4. Analysis
Calculations were made in terms of
percentage by using the following formula

114

2.4.1. Colonization frequency (CF %)

Individual fungal colonies that appeared in
the bait samples
CF % =

100
Total number of colonized segments

2.4.2. Relative percentage occurrence (RPO %)

The distribution and percentage of
occurrence of different groups of fungi was
calculated by using the following formula.
Density of colonization of single group
RPO % =

X 100
Total density of colonization

2.5. Enzyme activity of selected test fungi

The enzyme activity of the selected test
fungi were carried out according to the procedure
given by Chamier, 1985).
2.5.1. Amylolytic activity
Glucose Yeast Peptone (GYP) medium
with 0.2% soluble starch with pH 6 was used.
After 3 to 5 days of colony growth, the plates were
flooded with iodine solution. A yellow zone
around the fungal colony in an otherwise blue
medium indicated amylolytic activity.
2.5.2. Laccase activity
Glucose Yeast Peptone (GYP) medium
with 0.05 g, 1 naphtol/L with pH 6 was used. As
the fungus grows, the colourless medium turned
blue due to the oxidation of 1 naphtol by laccase.
2.6. In vitro antifungal activity of five medicinal
plants against test fungi
Fresh plant/plant parts were collected
randomly from S.D.N.B. Vaishnav College,
Chrompet, Chennai - 44.
The details of
plant/plant parts screened, their families,
vernacular names and their therapeutic uses are
given in Table - 2. Fresh plant materials were
washed under running tap water, air dried and then
homogenized to fine powder and stored in airtight
bottles. The air dried and powdered plant material
(10 g of each) was extracted with 100 ml of
methanol, kept on rotary shakes for 24 hrs and it
was filtered and centrifuged at 5000 rpm for 15

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

min. The supernatant was collected and

evaporated to dryness to give the crude dried
extract which was used for anti fungal assay (Jigna
Parekh and Sumitra Chanda, 2008).

Table - 1: List of location and kind of water

sample collected during the course of study
S. No

2.7. Antifungal assay

To evaluate the antifungal activity, sterile
agar plates were used according to the disc
diffusion assay. The test fungi were inoculated at
the centre of the agar plates. The sterile filter
paper discs (0.5 mm diameter) were impregnated
with plant extracts dissolved in methanol and
dried. Five individual discs of different plant
extracts were placed around the inoculated test
fungi and incubated at 28C for 48 hrs. Following
an incubation period of 4 days, the plates were
removed from the incubator and antifungal
activity was evaluated by measuring zones of
inhibition of fungal growth. Clear zones within
which fungal growth was absent were measured
and recorded as the diameter (mm) of complete
growth inhibition. Blank disc impregnated with
solvent methanol followed by drying off was used
as control. The whole experiment was performed
by making 0.5 cm wells on agar plates around the
test fungi and the wells were filled with 2 ml plant
extracts in DMSO (Dimethyl Sulphoxide) for
comparison with the disc diffusion method.

115

Kind
sample

of

Place of collection

1.

Lake water

Erumaiyur

2.

Lake water

Poonamallee

3.

Canal Water

Thiruneermalai

4.

Canal Water

Kundrathur

5.

Pond Water

Pallavaram

6.

Pond Water

Krishna Nagar

7.

Pond Water

Pazanthandalam

8.

Pond Water

Pallavaram

9.

Tap Water

S.D.N.B. Vaishnav
College, Chrompet

10.

Tap Water

Pallavaram

Table - 2: List of selected medicinal plants and their therapeutic uses

S. No.

Host plants

Common Name

Tamil Name

Medicinal Uses

1.

Melia azedarach L.

Neem tree

Vembu

2.

Boerhavia diffusa
Linn.
Lantana camara
Linn.

Pular

Punarnava

Anthelmintic,
antifungal,
antidiabetic,
antibacterial,
antiviral,
contraceptive
and
sedative
Scabies, myalgia, aphrodisiac

Red Sage

Mukkarattai

Plumeria rubra
Linn.
Ricinus communis
Linn.

Common
Frangipani
Castor oil plant

Champige tree

3.

4.
5.

Aamanakku

Antipyretic,
carminative,
Antidote to snake venom,
treatment of malaria, wounds,
cuts, ulcers, eczema, and tumours
Ulcers, leprosy, inflammations,
rube facient
Antimicrobial,
antihistamine,
Anti-inflammatory treatment of
jaundice and sores.

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

3. Results and Discussion

The widespread occurrence of aquatic
fungi present in the freshwater ecosystem has been
gained recognition only rather recently. The
present study has been aimed towards isolation
and identification of fungi associated with ten
different water samples collected from different
locations. Observations were made to various
parameters as mentioned in materials and
methods.
The hydrochemical analysis of ten
different water samples revealed that the lake
water samples of Poonamallee and Erumaiyur
(code no. 34659 and 34660) showed high level of
Ammonia. This indicates that the water is
extraneously polluted and the water sample is
chemically not potable. Similarly, the canal water
(code no. 34657 and 34664) collected from
Thiruneermalai and Kundrathur showed high level
of ammonia and nitrate value, which exceeds the
maximum allowable limit and the water samples
are chemically not potable. Among the four pond
water samples analyzed, two samples from
Erumaiyur and Pazanthandalam (code no. 34662
and 31872) was found to have more iron and
nitrate content and turbidity value exceeded the
maximum allowable limit and the water samples
were chemically not potable whereas the
remaining two pond samples (code no. 34663 and
34665)
collected
from
Pallavaram
and
Krishnanagar and two more tap water samples
collected from S.D.N.B. Vaishnav college for
Women, Chromepet and Pallavaram were found to
be potable.
Further, the water analysis of ten samples
indicates that the pH range was between 6.9 7.9
and it has almost all the nutrients namely calcium,
magnesium, sodium, potassium, iron, manganese,
free ammonia, nitrate, nitrite, chloride, fluoride,
sulfate and phosphate which supports growth of
the aquatic microorganism in natural environment.
A total of 1456 fungal colonies were obtained
using seeds as bait. These colonies were classified
into 33 species of fungi belonging to 20 genera (1
Oomycetes, 2 Zygomycetes, 2 Ascomycetes and
15 Hyphomycetes) and 13 non-sporulating sterile
morphospecies. The most frequently isolated

116

fungi using seeds as bait from ten experimental

water samples during the study period were
Pythium sp., Mucor sp., Rhizopus sp, Chaetomium
globosum, Alternaria alternata, Cladosporium
cladosporioides,
Fusarium
oxysporum,
Nigrospora sphaerica and Trichoderma viride.
The total numbers of fungi recovered from the
experimental samples using seeds as bait are
presented in Figure 1a.
Of the 983 fungal colonies recovered, 20
taxa (2 Zygomycetes, 2 Ascomycetes and 14
Hyphomycetes and 2 Coelomycetes) and 5 nonsporulating sterile morphospecies were recorded.
The commonly encountered fungi using plant
segments as bait during the study period were
Chaetomium gracile, Alternaria alternate,
Aspergillus flavus, Cladosporium cladosporioides,
Geomyces sp., Monilia sp., Penicillium citrinum,
Pestalotiopsis sp., and Colletotrichum sp. The
total numbers of fungal colonies from ten water
samples are presented in Figure 1b.
In this study of the 687 colonies recovered,
16 genera (2 Zygomycetes, 2 Ascomycetes, 2
Saccharomycetes and 10 Hyphomycetes) and 2
non-sporulating sterile morphospecies were
identified. The fungi namely Saccharomycopsis
sp.,
Brettanomyces
sp.,
Acremonium,
Paecilomyces sp. and Penicillium oxalicum were
more frequently isolated from the ten experimental
water samples using insects as bait. The
emergences of fungal colony from ten water
samples are presented in Figure 1c.
Altogether 450 fungal colonies (including
non-sporulating colonies) were recorded from the
ten experimental water samples using aquatic
animal segments as bait (Figure 1d).
The
colonies were classified under 13 genera (2
Saccharomycetes,
1
Ascomycetes,
10
Hyphomycetes) and 2 non-sporulating sterile
morphospecies.
The
fungi
such
as
Saccharomycopsis, Acremonium sp., Trichoderma
viride and Verticillium sp. were repeatedly
recovered during this study.
A total of 1038 fungal colonies belonging
to 22 genera were isolated from water samples
using fruits and vegetables as bait (Figure 1e).
The fungal composition included 2 Zygomycetes,

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

2
Ascomycetes,
17
Hyphomycetes,
1
Coelomycetes and 5 non-sporulating sterile
morphospecies. The most frequently isolated
fungi from ten water samples were Mucor sp.,
Rhizopus stolonifer, Curvularia lunata, Fusarium
oxysporum, Geomyces sp., Humicola sp. and
Verticillium sp.

117

A total of 1456 fungal colonies were

recorded from ten test samples using seeds as bait.
The seeds differ in their effectiveness as baits for
fungi associated with water. Among the five seed
baits tested seasame seeds trapped maximum
number of fungal colonies (361) followed by
pepper (304), chilli (285), cumin (277), and green
gram (229). Some of the seeds belong to starchy,
oily and proteinous food, hence their suitability as
bait may be due to their food content, which may
be comparable with the food requirements of the
fungi isolated. The effectiveness may also be
attributed to the kind of seeds used as well as the
exposed nature of the endosperm of the seeds
(Sparrow 1960).

A total of 4614 fungal colonies were

recorded. These colonies were classified into 27
species of fungi belonging to 23 genera (1
oomycetes, 2 zygomycetes, 1 ascomycetes, 2
saccharomycetes, 16 hyphomycetes and 2
coelomycetes) and the remaining 27 colonies were
classified under sterile morphospecies. The list of
fungi enumerated from water samples using
different techniques are presented in Table - 3.

Likewise, the total number of colonies

observed was maximum in plant twigs (225)
among the plant baits tested. Similarly, ant bait
trapped more fungal colonies (245) when
compared to the rest of the insect baits. Among the
animal segments tested for the bait fins showed
maximum number (131) of colonies. Out of the
fruits and vegetable baits tested more number of
colonies were observed in carrot (236) followed
by apple, beetroot, lemon and potato.

The ecological differences in different

geographical locations play an important role in
the species diversity of the fungi. In the present
investigation, the relative percentage occurrence
of different groups of fungi from ten experimental
samples showed the percent contribution of
hyphomycetes was very high followed by
zygomycetes, sachharomycetes, ascomycetes,
sterile
morphospecies,
coelomycetes
and
oomycetes (Figure - 2).

Figure 1a: Total number of fungi recovered from the experimental samples using seeds as bait

Total number of fungi recovered from the experimental samples using seeds as a bait

250

201

Total number of fungal isolates

184
200

172
147

160

164

140
127

150

101
100
60

50

0
Lake water
sample 1

Lake water
sample 2

Canal water Canal water

sample 1
sample 2

Sesame

Pepper

Pond water
sample 1

Chilli

Pond water
sample 2

Cumin

Pond water
sample 3

Pond water
sample 4

T ap water
sample 1

T ap water
sample 2

Green gram

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

118

Figure 1b: Total number of fungi recovered from the experimental samples using
plant segments as bait
Total number of fungi recovered from the experimental samples using plant segments as a bait
129
140
121

119

119

Total number of fungal isolates

120
99

95

92

97

100

80
62

50
60

40

20

0
Lake water
sample 1

Lake water
sample 2

Canal water
sample 1

Canal water
sample 2

Plant twigs

Grass

Pond water
sample 1
Leaf litter

Pond water
sample 2

Pond water
sample 3

Nymphea petiole

Pond water
sample 4

T ap water
sample 1

T ap water
sample 2

Woody materials

Figure 1c: Total number of fungi recovered from the experimental samples
using insects as bait
Total number of fungi recovered from the experimental samples using insects as a bait
114

Total number of fungal isolates

120

91

100

86

80

69

69

64
55
49

60

47

43

40

20

0
Lake w ater
sample 1

Lake w ater Canal w ater Canal w ater Pond w ater

sample 2
sample 1
sample 2
sample 1
Ant

Housefly

Pond w ater
sample 2

Pond w ater
sample 3

Pond w ater
sample 4

Tap w ater
sample 1

Tap w ater
sample 2

Wings of insects

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

119

Figure 1d: Total number of fungi recovered from the experimental samples
using animal segments as bait
Total number of fungi recovered from the experimental samples using animal segments as a bait
69
70

61

53

Total number of fungal isolates

60

49

46

46
50

42
40

29

28

27

30

20

10

0
Lake w ater
sample 1

Lake w ater
sample 2

Canal w ater Canal w ater

sample 1
sample 2
Fish scales

Pond w ater
sample 1
Gills

Pond w ater
sample 2
Fins

Pond w ater
sample 3

Pond w ater
sample 4

Tap w ater
sample 1

Tap w ater
sample 2

Fish t ails

Figure 1e: Total number of fungi recovered from the experimental samples
using fruits and vegetables as bait
Total number of fungi recovered from the experimental samples using fruits and vegetables as a bait
134
129

140

120

125

114

117

91

90

100

80

56

62

60

40

20

0
Lake w ater
sample 1

Lake w ater
sample 2

Canal w ater
sample 1

Canal w ater
sample 2
Apple

Pond w ater
sample 1

Lemon

Potato

Pond w ater
sample 2
Carrot

Pond w ater
sample 3

Pond w ater
sample 4

Tap w ater
sample 1

Tap w ater
sample 2

Beetroot

Figure 2: Percent contribution of different groups of fungi

Percent contribution of different groups of fungi associated with water from
selected baits

Fr uits and ve ge table s

Anim al s e gm e nts

Ins e cts

Plant s e gm e nts

Sterile Morphospecies

Coelomycetes

Hyphomycetes

Sachharomycetes

Ascomycetes

Zygomycetes

Se e ds

Oomycetes

Total number of fungal isolates

120

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Aspergillus sp., and Penicillium sp., have been

previously reported in the bathing sites of the
river, Poland (Bozena Kiziewicz, 2004). In this
study, the experimental baits such as aquatic plant
twigs, grass, woody materials and vegetables like
carrot, beetroot act as a good nutritive source for
the isolation of fungi like Humicola sp., Monilia
sp., Acremonium sp., Gliocladium sp., Fusarium
sp., Colletotrichum sp., and Verticillium sp.
Bozena Kiziewicz and Alicja Kurzatkowska 2004
have remarked that the fungi associated with the
water samples can be trapped by using animal
segments as bait. In this study, the genera
Saccharomycopsis and Brettanomyces were
isolated more frequently by using insects as bait.
The fungi Pythium, which is reported here has
already been recorded as an aquatic fungi (Bozena
Kiziewicz, 2004).

Table - 3: List of fungi enumerated from the

water sample during the course of study
S. No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27

120

List of Isolated fungi

OOMYCETES
Pythium sp.
ZYGOMYCETES
Mucor sp.
Rhizopus stolonifer
ASCOMYCETES
Chaetomium globosum
Chaetomium gracile
SACCHAROMYCETES
Saccharomycces sp.
Brettanomyces sp.
HYPHOMYCETES
Acremonium sp.
Alternaria alternate
Aspergillus flavus
A. fumigates
A. niger
A. terreus
Cladosporium cladosporioides
Curvularia lunata
Curvularia tuberculata
Drechslera halodes
Fusarium oxysporum
Geomyces sp.
Gliocladium roseum
Humicola sp.
Nigrospora sphaerica
Penicillium citrinum
Penicillium oxalicum
Trichoderma viride
COELOMYCETES
Colletotrichum sp.
Pestalotiopsis sp.

Most of the fungal biomass on decaying

leaves consists of vegetative hyphae that cannot be
identified through conventional microscopy
(Nikolcheva et al., 2003). Aquatic hyphomycetes,
an artificial phylogenitically, heterogenous group
of true fungi are fungi dominating leaf
decomposition in streams. They are anamorphs of
Ascomycota and Basidiomycota (Alexopoulos et
al., 1996). The presence of sterile forms continues
to frustrate the mycologists because of their
uncertain taxonomy. Moreover, they demand the
use of molecular techniques for classification
(Bills, 1996). The nonsporulating sterile forms
recovered during the current study were separated
into culture groups based on the colony
morphology, hyphal mat characteristics (texture,
zonation), presence of sclerotia and pigmentation
as described by Frohlich et al. (2000).
Apart from trying to understand the
biology of fungi associated with water another
motivation to study these fungi is its ability to
produce enzyme. Farida et al. (2001) reported
that, most of the aquatic fungi produced amylase,
cellulase, pectinase, protease, lipase and xylanase.

The most commonly encountered fungi in

various ecosystems namely Mucor sp., Rhizopus
sp., Chaetomium sp., Saccharomyces sp.,

These enzymes may cause breakdown of

leaf tissues and increase the palatability of the
leaves to leaf eating invertebrates (Barlocher,
1992; Suber Kropp, 1992).
In the present
investigation, amylase activity was observed in

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

almost all the test fungi. Among the 11 fungi

tested, one test fungi namely Trichoderma viride
showed that laccase activity which has been
previously reported (Zaldivar et al., 2001).

121

Aquatic and soil environments play a

significant part in the process of mycotic
infections. A similar study of screening natural
plant extracts against different fungal pathogens
was well recorded in literature (Banso and
Adeyemo, 2007). In this study, attempts were
made to investigate the inhibitory effects of
organic solvent extracts from 5 medicinal plant
species against 10 test fungi (Table - 4) to evaluate
the potential application of medicinal plant based
treatments to control disease caused by pathogenic
fungi. Results revealed that extracts of Boerhavia
diffusa, Lantana camara and Ricinus communis
can be used as a potent biocide to treat diseases in
plants.

In general, plants generally produce many

secondary metobolites which constitute an
important source of microbicides, pesticides and
many pharmaceutical drugs (Satish et al., 2008;
Vetrivel Rajan et al., 2009). Although most of the
fungi associated with water have been reported to
play an important role in decomposition and
bioremediation, certain fungi can cause diseases in
plants and animals. Their spores infect plants
most frequently perforating the tissue that covers
stems, leaves, fruits and roots.

Table - 4: Antifungal activities of the test fungi

Antifungal activities of the test fungi
S.
No.

Methanol extracts of selected medicinal plants

Name of the test fungi

Zone of inhibition (mm)

Melia
azedarach

Boerhavia
diffusa

Lantana
camara

Plumeria
rubra

Ricinus
communis

Alternaria alternate

10

Aspergillus flavus

14

10

16

Cladosporium
cladosporioides

14

11

Curvularia lunata

Drechslera halodes

11

Fusarium oxysporum

Penicillium oxalicum

12

11

10

Pestalotiopsis sp.

12

Rhizopus stolonifer

10

Trichoderma viride

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V. Bhuvaneswari/ Life Science Archives (LSA), Volume 1, Issue 2, Page 112 to 123, 2015

122

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