Pt.

RAVISHANKAR UNIVERSITY RAIPUR (CHHATTISGARH)

SEMINAR ON

GUIDED BY DR. K. RAMASWAMY FPIC, CFTRI PRANITA SHARMA FIVE YEAR INTEGRATED X SEMESTER MICROBIOLOGY SOS IN LIFE SCIENCE

Introduction History Objectives Materials and Method Results Discussion Summary References

INTRODUCTION

INTRODUCTION
The term filamentous fungi is used for species producing filament like hyphae. E.g. Essentially unicellular fungi with vegetative cells capable of repeated budding.

The development of antifungal agents can be useful in the control of infections caused by Phytopathogenic fungi.

Secondary metabolites - Storage and defense mechanism. Not necessary for their development and growth.

HISTORY

HISTORY
Originally noticed by a French medical student, Ernest Duchesne, in 1896. Penicillin was re-discovered by bacteriologist Alexander Fleming working at St. Mary's Hospital in London in 1928.

On May 25, 1948, Andrew J Moyer was granted a patent for a method of the mass production of penicillin.

OBJECTIVES

OBJECTIVE OF THE PRESENT STUDY

Isolation, screening and identification from food grains and soil samples.

Production of secondary metabolites in antibiotic test broth (ATB). Extraction and separation of crude metabolites using standard methods.
Testing (fungicidal activity) of crude metabolites against some plant pathogenic fungi (Aspergillus flavus, Colletotrichum sp., C.paradoxa, Fusarium sp., Penicillium rubrum, Rhizopus sp.). Testing for some human pathogenic bacteria (antibacterial activity) (Escherichia coli, Listeria monocytogenes, Micrococcus leuteus, Staphylococcus aureus).

MATERIALS AND METHOD

MATERIALS AND METHOD

Source of soil sample

Totally 24 different soil samples were collected from three different locations in and around Mysore city (Karnataka) which are: i. CFTRI Campus ii. Mysore University iii. Kukkarahalli Lake

Isolation of fungi from soil and other sources Identification of fungi isolated Isolation of pure culture Preliminary test against fungal pathogens Preparation of fungal spore inoculum (0.1% Tween 80) Transferred to ATB Extraction of crude metabolite Antifungal activity test Antibacterial activity test Identification assay for the most active fungal compound Thin Layer Chromatography Column Chromatography Chemical analysis (infra red spectra)

RESULTS

RESULTS
Different mycobiota of sample collected sites
S.No. Fungal genera S1 S2 S3 S4 Studied Samples (+, Present ; -, Absent) S5 Grains Leaf Vegetable

01
02 03 04 05 06 07

Alternaria sp.
Aspergillus sp. Ceratocystis sp. Cladosporium sp. Colletotrichum sp. Fusarium sp. Helminthosporium sp.

+ + + -

+
+ + + + + +

+
+ + + +

+ + + + -

+ + -

+ +

+
+ + + +

+ + + + -

08 09 10 11

Nigrospora sp. Penicillium sp. Rhizopus sp. Syncephalastrum sp.

+ + + -

+ + -

+ -

+ + +

+ + + +

+ + -

+ + + +

+ + -

12

Torula sp.

 Soil S2 was richest in fungal colony whereas S5 contains least number of fungus. Grains Few fungi were found in grains e.g. Aspergillus sp., Helminthosporium sp. Leaf Various kinds of plant pathogenic fungi was seen on leaf isolates e.g. AlternariaI sp., Aspergillus sp., Cladosporium sp., Nigrospora sp. and Torula sp.

Vegetable It was also rich in fungal isolates.

Preliminary Antimicrobial activity of some Penicillium isolates against plant pathogenic fungi Zone of Inhibition Isolates No. A.flavus Colletotrichum C.paradoxa P.rubrum P.soppi _ + + _ _ _ Fusarium Rhizopus P.rubrum sp. sp. + _ _ _ _ _

P.brevicampactum
P2 P19 P20 P21 P22 P24 P27 P2(N)

+
+ _ _ _ _ _ + +

+
_ _ + + _ + _ +

+
+ + + + _ + _ +

+
+ _ + + + + _ +

_
_ _ _ _ _ _ _ _

_
_ _ _ _ _ _ _ _

Thin Layer Chromatography

Column Chromatography

UV Spectra of P. rubrum

UV Spectra of P2

SUMMARY

Soil Isolates

Pure Culture of Penicillium

Preliminary Test

Antibiotic Test Broth

REFERENCES

REFERENCES
Chutia, M. and Ahmed, G.U. (2012): Diversity, antimicrobial activities and associated microbiota of soil Penicillia from Virgin Forest Floor. Journal of Microbiology, Biotechnology and Food Sciences, 5, 1279-1294. Dayalan, S.A.J., Darwin, P., Prakash, S., (2011): Comparative study on production, purification of penicillin by Penicillium chrysogenum isolated from soil and citrus samples. Asian Pacific Journal of Tropical Biomedicine, 15-19. Devi, P. DSouza, L., Kamat, T., Rodrigues, C. and Naik, C.G. (2009): Batch culture fermentation of Penicillium chrysogenum and a report in the isolation, purification, identification and antibiotic activity of citrinin. Indian Journal of Marine Sciences, 38, 38-44. Fleming, A. (1929): On the antibacterial action of cultures of a Penicillium sp., with special reference to their use in the isolation of B. influenzae. British Journal of Experimental Pathology, 10, 226-236. Khaddor, M., Saidi, R., Aidoun, A., Lamarti, A., Elaraki, A.T., Ezziyyani, M., Castillo, M.E.C. and Badoc, A. (2007): Antibacterial effects and toxigenesis of Penicillium aurantiogriseum and P. viridicatum. African Journal of Biotechnology, 6, 2314-2318. Zhelifonova, V. P., Antipova, T. V., Ozerskaya, S. M., Kochkina, G. A., and Kozlovsky, A. G. (2008): Secondary Metabolites of Penicillium Fungi Isolated from Permafrost Deposits as Chemotaxonomic Markers. Microbiology, 78, 350354.

SEARCH ENGINE
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