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Available online at www.jpsscientificpublications.com

Life Science Archives LSA)

ISSN: 2454-1354

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

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Research Article

I n vitroSTUDIES OF ANTIFUNGAL AND ENZYMATIC ACTIVITY OF

SELECTED FUNGI ISOLATED FROM WATER USING DIFFERENT BAITS

S.A. Aghizion Inbakani1, V. Bhuvaneswari*

2, G. Kathiravan

3and B. Shanmugapriya

2

1S.D.N.B. Vaishnav College for Women, Chromepet, Chennai - 600 044, Tamil Nadu, India.

2Chikkaiah Naicker College, Veerapan chitram, Erode 600 004, Tamil Nadu, India.

3Department of Biotechnology ,Vels University, Pallavaram, Chennai600 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 aredestructive 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 individualgroups 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 methodto 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.Ou

of the methanol extracts of the five medicinal plants tested, Boerhavia diffusa, Lantana camara,andRicinus

communis showed best antifungal activity against Aspergillus flavus, Cladosporium cladosporioides, andDrechslera 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.

ArticleHistoryReceived : 21.03.2015Revised : 01.04.2015

Accepted : 06.04.2015

Key words Aquatic fungi, Enzyme activityMedicinal plants and Plant extract.

1. Introduction

Fungi are universally present in all typesof 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: [email protected]

Several baits autoclaved wood cubes, filter paper

strips, dead fishes, meat piece, dead flies, insect

larvae, cooked egg white, and discs of cork havebeen used to isolate aquatic fungi (Alabi, 1971

Sharp 1978; Agina and Kpu, 1988). The use of

hemp seeds as baits (Sparrow, 1960; Lui and Volz1977; 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; Roberts1963; 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 113


pears are very good as baits for aquatic

phytophthora species (Adenle, 1994). The aim of

this study is to access the suitability of differentkinds 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 ofthe 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 activityof methanol extracts of some Indian medicinal

plants against test fungi was done by agar disc

diffusion method to evaluate its potentialimportance.

2. Methodology

General microbiological and laboratorytechniques followed in the present investigation

were as there outlined by Booth (1971). In thepresent 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 aroundChennai, 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 collectedand used for further experimentation. The list of

location and kind of water samples are presentedin 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 WaterTesting 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, calciumcarbonate, and silica.

2.1. Methodology for recovery of fungi

associated with water samples

Isolation of fungi was carried out by using

the following isolation techniques viz.,

(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 inpetriplates. 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 containingculture media. The following culture media

namely, Water Agar Medium (WAM), PotatoDextrose 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 litrecontainers and covered properly to protect thewater from penetration by bacteria. After a few

days of incubation, microscopically determined

mycelia were removed from the seeds and

transferred to culture media amended withchloramphenicol 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 animalsegments, samples were collected from the same

location and washed thoroughly under tap water toremove the debris and surface sterilized with 1%

mercuric chloride for 10 min and rinsed with

distilled water for about 5 min and then finallydried on a sterilized filter paper. The surface

sterilized segments were platted on culture media

for the enumeration of fungi.


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2.2. Methodology for recovery of fungi

associated with mud samples

The following baiting technique was used

for the recovery of fungi from mud samples (El-Hissy and Abal-Eloah, 1989). A 50 g of each mud

sample was introduced into clean sterilepetridishes. The mud sample in each petridish was

then flooded with sterile distilled water withaddition of few sesame seeds. The submerged mud

samples were also processed using certain

vegetables as bait. In this method, the selectedvegetable 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 coveredwith 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 forthe 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 wereremoved as soon as they appeared on the plates.

The identification of fungi was based on

morphological features such as shape and size ofhyphae, 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 ofpercentage by using the following formula

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 ofoccurrence 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 proceduregiven 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 wereflooded 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. Asthe fungus grows, the colourless medium turned

blue due to the oxidation of 1 naphtol by laccase.

2.6. I n vitroantifungal activity of five medicinal

plants against test fungi

Fresh plant/plant parts were collectedrandomly from S.D.N.B. Vaishnav College

Chrompet, Chennai - 44. The details ofplant/plant parts screened, their familiesvernacular 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 airtightbottles. 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 itwas filtered and centrifuged at 5000 rpm for 15


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min. The supernatant was collected and

evaporated to dryness to give the crude dried

extract which was used for anti fungal assay (JignaParekh and Sumitra Chanda, 2008).

2.7. Antifungal assay

To evaluate the antifungal activity, sterileagar plates were used according to the disc

diffusion assay. The test fungi were inoculated at

the centre of the agar plates. The sterile filterpaper discs (0.5 mm diameter) were impregnated

with plant extracts dissolved in methanol and

dried. Five individual discs of different plantextracts 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 ofinhibition of fungal growth. Clear zones within

which fungal growth was absent were measuredand 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 performedby 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) forcomparison with the disc diffusion method.

Table - 1: List of location and kind of water

sample collected during the course of study

. NoKind of

samplePlace 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 WaterS.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 azedarachL. Neem tree Vembu Anthelmintic, antifungal,antidiabetic, antibacterial,

antiviral, contraceptive and

sedative

2. Boerhavia diffusa

Linn.

Pular Punarnava Scabies, myalgia, aphrodisiac

3. Lantana camara

Linn.

Red Sage Mukkarattai Antipyretic, carminative,

Antidote to snake venom,treatment of malaria, wounds,cuts, ulcers, eczema, and tumours

4. Plumeria rubraLinn.

CommonFrangipani

Champige tree Ulcers, leprosy, inflammations,rube facient

5. Ricinus communis

Linn.

Castor oil plant Aamanakku Antimicrobial, antihistamine,

Anti-inflammatory treatment ofjaundice and sores.
http://en.wikipedia.org/wiki/Anthelmintichttp://en.wikipedia.org/wiki/Antidiabetichttp://en.wikipedia.org/wiki/Antibacterialhttp://en.wikipedia.org/wiki/Antiviral_drughttp://en.wikipedia.org/wiki/Contraceptivehttp://en.wikipedia.org/wiki/Sedativehttp://en.wikipedia.org/wiki/Antipyretichttp://en.wikipedia.org/wiki/Carminativehttp://en.wikipedia.org/wiki/Carminativehttp://en.wikipedia.org/wiki/Antipyretichttp://en.wikipedia.org/wiki/Sedativehttp://en.wikipedia.org/wiki/Contraceptivehttp://en.wikipedia.org/wiki/Antiviral_drughttp://en.wikipedia.org/wiki/Antibacterialhttp://en.wikipedia.org/wiki/Antidiabetichttp://en.wikipedia.org/wiki/Anthelmintic


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3. Results and Discussion

The widespread occurrence of aquatic

fungi present in the freshwater ecosystem has been

gained recognition only rather recently. Thepresent study has been aimed towards isolation

and identification of fungi associated with tendifferent water samples collected from different

locations. Observations were made to variousparameters as mentioned in materials and

methods.

The hydrochemical analysis of ten

different water samples revealed that the lakewater 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 levelof 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 fromErumaiyur and Pazanthandalam (code no. 34662

and 31872) was found to have more iron and

nitrate content and turbidity value exceeded themaximum allowable limit and the water samples

were chemically not potable whereas theremaining two pond samples (code no. 34663 and

34665) collected from Pallavaram andKrishnanagar and two more tap water samples

collected from S.D.N.B. Vaishnav college for

Women, Chromepet and Pallavaram were found tobe 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 and15 Hyphomycetes) and 13 non-sporulating sterile

morphospecies. The most frequently isolated

fungi using seeds as bait from ten experimental

water samples during the study period were

Pythium sp.,Mucorsp.,Rhizopussp, Chaetomium

globosum, Alternaria alternata, Cladosporiumcladosporioides, Fusarium oxysporum

Nigrospora sphaerica and Trichoderma viride

The total numbers of fungi recovered from theexperimental samples using seeds as bait are

presented in Figure 1a.

Of the 983 fungal colonies recovered, 20

taxa (2 Zygomycetes, 2 Ascomycetes and 14Hyphomycetes and 2 Coelomycetes) and 5 non-

sporulating sterile morphospecies were recorded

The commonly encountered fungi using plant

segments as bait during the study period wereChaetomium gracile, Alternaria alternate

Aspergillus flavus, Cladosporium cladosporioides

Geomyces sp., Monilia sp., Penicillium citrinum

Pestalotiopsis sp., and Colletotrichum sp. Thetotal 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, 2Saccharomycetes and 10 Hyphomycetes) and 2

non-sporulating sterile morphospecies were

identified. The fungi namely Saccharomycopsissp., Brettanomyces sp., Acremonium

Paecilomyces sp. and Penicillium oxalicum weremore frequently isolated from the ten experimental

water samples using insects as bait. Theemergences of fungal colony from ten water

samples are presented in Figure 1c.

Altogether 450 fungal colonies (including

non-sporulating colonies) were recorded from theten 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 sterilemorphospecies. The fungi such as

Saccharomycopsis, Acremoniumsp., 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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184

201

147

127

140

172160 164

60

101

0

50

100

150

200

250

Totalnum

beroffungalisolate

Lake water

sample 1

Lake water

sample 2

Canal water

sample 1

Canal water

sample 2

Pond water

sample 1

Pond water

sample 2

Pond water

sample 3

Pond water

sample 4

Tap water

sample 1

Tap water

sample 2

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

Sesame Pepper Chilli Cumin Green gram

2 Ascomycetes, 17 Hyphomycetes, 1

Coelomycetes and 5 non-sporulating sterile

morphospecies. The most frequently isolatedfungi from ten water samples were Mucor sp.,

Rhizopus stolonifer, Curvularia lunata,Fusarium

oxysporum, Geomyces sp., Humicola sp. and

Verticillium sp.

A total of 4614 fungal colonies wererecorded. These colonies were classified into 27

species of fungi belonging to 23 genera (1

oomycetes, 2 zygomycetes, 1 ascomycetes, 2saccharomycetes, 16 hyphomycetes and 2

coelomycetes) and the remaining 27 colonies were

classified under sterile morphospecies. The list of

fungi enumerated from water samples usingdifferent techniques are presented in Table - 3.

The ecological differences in differentgeographical locations play an important role in

the species diversity of the fungi. In the presentinvestigation, the relative percentage occurrence

of different groups of fungi from ten experimental

samples showed the percent contribution of

hyphomycetes was very high followed byzygomycetes, sachharomycetes, ascomycetes,

sterile morphospecies, coelomycetes and

oomycetes (Figure - 2).

A total of 1456 fungal colonies were

recorded from ten test samples using seeds as bait

The seeds differ in their effectiveness as baits forfungi 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 greengram (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 thefungi 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).

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) whencompared to the rest of the insect baits. Among the

animal segments tested for the bait fins showed

maximum number (131) of colonies. Out of thefruits and vegetable baits tested more number of

colonies were observed in carrot (236) followed

by apple, beetroot, lemon and potato.

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


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119

99

119 121

9295

129

97

5062

0

20

40

60

80

100

120

140

Totalnumberoffungalisolate

Lake water

sample 1

Lake water

sample 2

Canal water

sample 1

Canal water

sample 2

Pond water

sample 1

Pond water

sample 2

Pond water

sample 3

Pond water

sample 4

Tap water

sample 1

Tap water

sample 2

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

Pla nt t wi gs Grass L ea f li tt er Nym phea pe tio le W oody ma te ria ls

91

114

69

86

6964

4943

55

47

0

20

40

60

80

100

120

Totalnumberoffungalisolates

Lake water

sample 1

Lake water

sample 2

Canal w ater

sample 1

Canal w ater

sample 2

Pond w ater

sample 1

Pond w ater

sample 2

Pond w ater

sample 3

Pond w ater

sample 4

Tap water

sample 1

Tap water

sample 2

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

Ant Housefly Wings of insects

Figure 1b: Total number of fungi recovered from the experimental samples using

plant segments as bait

Figure 1c: Total number of fungi recovered from the experimental samples

using insects as bait


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2927

46

28

53

46

61

69

49

42

0

10

20

30

40

50

60

70


Lake water

sample 1

Lake water

sample 2

Canal water

sample 1

Canal water

sample 2

Pond water

sample 1

Pond water

sample 2

Pond water

sample 3

Pond water

sample 4

Tap water

sample 1

Tap water

sample 2

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

Fish scales Gills Fins Fish tails

120114

129

125

90 91

134

117

56 62

0

20

40

60

80

100

120

140


Lake water

sample 1

Lake w ater

sample 2

Canal w ater

sample 1

Canal w ater

sample 2

Pond w ater

sample 1

Pond w ater

sample 2

Pond w ater

sample 3

Pond w ater

sample 4

Tap water

sample 1

Tap water

sample 2

Total number of fungi recovered from the experimental samples using fruits and vegetables as a bait

Apple Lemon Potato Carrot Beetroot

Oomycetes

Zygomycetes

Ascomycetes

Sachharomycetes

Hyphomycetes

Coelomycetes

SterileMorphospecies

Seeds

Plant segm ents

Insects

Animal segments

Fruits and vegetables

Percent contribution of different groups of fungi associated with water from

selected baits

Figure 1d: Total number of fungi recovered from the experimental samples

using animal segments as bait

Figure 1e: Total number of fungi recovered from the experimental samples

using fruits and vegetables as bait

Figure 2: Percent contribution of different groups of fungi


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Table - 3: List of fungi enumerated from the

water sample during the course of study

The most commonly encountered fungi invarious ecosystems namely Mucor sp., Rhizopus

sp., Chaetomium sp., Saccharomyces sp.,

Aspergillus sp., and Penicillium sp., have been

previously reported in the bathing sites of the

river, Poland (Bozena Kiziewicz, 2004). In thisstudy, 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., Moniliasp., Acremonium sp., Gliocladium sp., Fusarium

sp., Colletotrichum sp., and Verticillium sp

Bozena Kiziewicz and Alicja Kurzatkowska 2004

have remarked that the fungi associated with thewater samples can be trapped by using animal

segments as bait. In this study, the genera

Saccharomycopsis and Brettanomyces wereisolated 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).

Most of the fungal biomass on decayingleaves consists of vegetative hyphae that cannot be

identified through conventional microscopy

(Nikolcheva et al., 2003). Aquatic hyphomycetesan artificial phylogenitically, heterogenous group

of true fungi are fungi dominating leaf

decomposition in streams. They are anamorphs of

Ascomycota and Basidiomycota (Alexopoulos eal., 1996). The presence of sterile forms continues

to frustrate the mycologists because of their

uncertain taxonomy. Moreover, they demand theuse of molecular techniques for classification

(Bills, 1996). The nonsporulating sterile forms

recovered during the current study were separated

into culture groups based on the colonymorphology, hyphal mat characteristics (texture

zonation), presence of sclerotia and pigmentation

as described by Frohlich et al. (2000).

Apart from trying to understand thebiology of fungi associated with water another

motivation to study these fungi is its ability toproduce enzyme. Farida et al. (2001) reported

that, most of the aquatic fungi produced amylase,cellulase, pectinase, protease, lipase and xylanase.

These enzymes may cause breakdown of

leaf tissues and increase the palatability of the

leaves to leaf eating invertebrates (Barlocher1992; Suber Kropp, 1992). In the present

investigation, amylase activity was observed in

S. No. List of Isolated fungi

OOMYCETES

1 Pythiumsp.

ZYGOMYCETES

2 Mucorsp.

3 Rhizopus stolonifer

ASCOMYCETES

4 Chaetomium globosum

5 Chaetomium gracile

SACCHAROMYCETES

6 Saccharomyccessp.

7 Brettanomycessp.HYPHOMYCETES

8 Acremoniumsp.

9 Alternaria alternate

10 Aspergillus flavus

11 A. fumigates

12 A. niger

13 A. terreus

14 Cladosporium cladosporioides

15 Curvularia lunata16 Curvularia tuberculata

17 Drechslera halodes

18 Fusarium oxysporum

19 Geomyces sp.

20 Gliocladium roseum

21 Humicola sp.

22 Nigrospora sphaerica

23 Penicillium citrinum

24 Penicillium oxalicum25 Trichoderma viride

COELOMYCETES

26 Colletotrichumsp.

27 Pestalotiopsissp.


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almost all the test fungi. Among the 11 fungi

tested, one test fungi namely Trichoderma viride

showed that laccase activity which has beenpreviously reported (Zaldivar et al., 2001).

In general, plants generally produce many

secondary metobolites which constitute animportant 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 andbioremediation, 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.

Aquatic and soil environments play a

significant part in the process of mycotic

infections. A similar study of screening naturaplant 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 oforganic 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 pathogenicfungi. Results revealed that extracts of Boerhavia

diffusa, Lantana camara and Ricinus communis

canbe used as a potent biocide to treat diseases inplants.

Table - 4: Antifungal activities of the test fungi

S.

No.

Antifungal activities of the test fungi

Name of the test fungi

Methanol extracts of selected medicinal plants

Zone of inhibition (mm)

Melia

azedarach

Boerhavia

diffusa

Lantana

camara

Plumeria

rubra

Ricinus

communis

1 Alternaria alternate 5 3 - 10 -

2 Aspergillus flavus 14 - 10 5 16

3Cladosporium

cladosporioides- 14 9 5 11

4 Curvularia lunata - 7 9 5 -

5 Drechslera halodes 11 9 7 - 8

6 Fusarium oxysporum - - - - 5

7 Penicillium oxalicum - 12 11 - 10

8 Pestalotiopsis sp. - - 12 - -

9 Rhizopus stolonifer - 3 5 - 4

10 Trichoderma viride - - - - 7


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