SCREENING AND TENTATIVE INDENTIFICATION OF TANNERY WASTE DEGRADING Group Members: Jothishwar . S – 15BBT0190 Keerthic Aswin. S- 15BBT0234 Kannan. C - 15BBT0235 Bhuvanesh .T -15BBT0236 ACTINOMYCES
SCREENING AND TENTATIVE INDENTIFICATION OF TANNERY WASTE DEGRADING
Group Members:Jothishwar . S – 15BBT0190Keerthic Aswin. S- 15BBT0234Kannan. C - 15BBT0235Bhuvanesh .T -15BBT0236
ACTINOMYCES
Objectives Of Our Project
• Cultivation of Actinomyces from the Soil Sample , Which is Taken from a Tannery industry in Ranipet.
• Isolating pure colonies of Actinomyces• Identification of Actinomyces ( Morphology & Staining)• Identifying Biochemical Activities of the culture
Actinomyces• It is a GRAM-POSITIVE bacteria.• Primarily , it is a Soil Bacteria.• Actinomyces Colonies form Fungus like Branches (hyphae)• Facultatively Anaerobic ; They form Endospores ;• Not easy to culture and Isolate• They have various Economical Values – Enzymes,
Antibiotics, Soil Ecology.
Collection of Soil Samples Ranipet Tannery Industry Effluents
Collecting Tannery Effluent & Leather Wastes• Add your first bullet point
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Cultivation of Actinomycets
Media We Used• Starch Casein Agar• Kenknight & Munaier's Medium• Actinomycetes Isolation Agar (INCUBATION PEROID – 8 Day degree celciuss at 30 – 35 degree celcius ))
Methods for Screening• Pour Plate Method
• streak Plate method
• Serial Dilution
Results for Third time Cultivation
Growth of Actinomycetes
First 2 times we did not get the colonies ( WHITE POWDER LIKE COLONIES)
Sub culturing- To Isolate the pure Colonies
Actinomyces Colonies
Streak plate Method
Starch Casein Media
Gram Staining - (identification)• Smear Preparation, Staining, decolourization was done.• Purple colour was Observed under Microscope.• It Proves that , it is a Gram + microorganism. • Also Filamentous Growth Of Colonies Were Observed
BIO CHEMICAL TESTBiochemical Test kit – Hi Media KB005A – Gram positive KB002 - Gram Negative rods
Broth Culture of Actinomyces
Biochemical Test - ResultsColour Changes were observed in the Rings of the kits
KB002
KB005A
BIOCHEMICAL TEST POSITIVE RESULTS
• Citrate utilization • Leucine utilization• Ornithine utilization• Urease• Phenylalanine deamination• Nitrate Reduction• Arginine Utilization
All other Biochemical test results are Negative only…
CONCLUSION• Staining and Biochemical tests proves that the Microorganism in the media is ACTINOMYCES
• These microbes are able to produce TANNASE enzyme
(Tannin acyl hydrolase) , Which will degrade the components present in tannery waste and effluents.
• We can go for Enzyme Activity Test , Which will give the results of degradation process.
• We have many microbes which helps us to protect the Environment. We Must know them and Use them in the Large scale
• Tannase enzyme is also having higher Economical Values
References• Bergey's Manual of Determinative Bacteriology• Screening of Tannase -Producing Fungi Isolated from Tannin-
Rich Sources ., Hamada A. Abou-Bakr*, Malak A. El-Sahn and Amr A. El-Banna
• Tannase Production By Aspergillus niger ., N. LOKESWARI* and K. JAYA RAJU.
ACKNOWLEDGEMENT• We would like to express our special thanks of gratitude to
our faculty, Prof.Dr.V. Suneetha ,who gave us the opportunity and Inspiration to do the project on Bioremediation topic which is also helped us in doing a lot of Research and We came to know about so many new things we are really thankful to her. Also we would like to Thank Research Scholars who gave us the helps in laboratories.