The document summarizes a student project aimed at isolating and identifying Actinomyces bacteria from soil samples taken from a tannery industry. The objectives were to cultivate Actinomyces, isolate pure colonies, identify morphology and staining, and determine biochemical activities. Samples were collected and Actinomyces was cultured using various media and methods over 8 days. Gram staining of isolated colonies showed purple, filamentous, Gram-positive bacteria. Biochemical tests identified abilities including citrate utilization and nitrate reduction. The conclusion was that Actinomyces was identified, which can degrade tannery waste through the tannase enzyme and potentially be used for bioremediation.

1. SCREENING AND
TENTATIVE INDENTIFICATION OF
TANNERY WASTE DEGRADING
Group Members:
Jothishwar . S – 15BBT0190
Keerthic Aswin. S- 15BBT0234
Kannan. C - 15BBT0235
Bhuvanesh .T -15BBT0236
ACTINOMYCES

2. 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

3. 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.

4. Collection of Soil Samples
Ranipet Tannery Industry Effluents

5. Collecting Tannery Effluent & Leather Wastes
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6. 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

7. Results for Third time Cultivation
Growth of
Actinomycetes
First 2 times we did not get the colonies ( WHITE POWDER LIKE COLONIES)

8. Sub culturing- To Isolate the pure Colonies
Actinomyces
Colonies
Streak plate Method
Starch Casein Media

9. 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

10. BIO CHEMICAL TEST
Biochemical Test kit – Hi Media KB005A – Gram positive
KB002 - Gram Negative rods
Broth Culture of Actinomyces

11. Biochemical Test - Results
Colour Changes were observed in
the Rings of the kits
KB002
KB005A

12. 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…

13. 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

14. 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.

15. 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.