This document presents research on the in vitro degradation of plastic via the microorganisms Micrococcus luteus and Masoniella species. Polystyrene plastic samples and soil samples were collected and various bacteria and fungi were isolated. M. luteus and Masoniella sp. were identified and found to degrade plastic at varying rates when incubated with the plastic for 35, 45, and 55 days. M. luteus degraded up to 32% of plastic weight while Masoniella sp. degraded up to 27.4% of plastic weight over the incubation periods. The study demonstrates that M. luteus and Masoniella sp. have potential for plastic biodegradation.
1. IN VITRO DEGRADATION OF PLASTIC
VIA
MICROCOCCUS LUTEUS AND
MASONIELLA SPECIES
• Presented by – GAURAV GANGWAR
2. INTRODUCTION
• WHAT IS PLASTIC?
- A polymer material which become mobile
on heating and thus can be cast into moulds.
-durable and slowly degradable.
• Adventage and Disadvantages
3. MATERIAL AND METHOD
• COLLECTION OF SAMPLE
1. POLYSTYRENE COLLECTION.
Collection of plastic cups from waste
disposal.
2. SOIL COLLECTION.
Any dumped area with large amount of
plastic waste disposal.
5. PROCESSING OF SOIL SAMPLE
• SERIAL DILUTION OF SOIL
• INOCULATION OF 10-5 AND 10-6 DILUTION –
BACTERIA ON NUTRITENT BROTH
• 10-2 AND 10-3 – FOR FUNGI ON PDA
6. INDENTIFICATION
• FOR BACTERIA
*GRAM STAINING
*MOTILITY TEST (HANGING DROP METHOD)
*BIOCHEMICAL TEST – 1.IMViC TEST
2. CATALASE TEST
3. OXIDASE TEST
• FOR FUNGI
* LECTOPHENOL COTTON BLUE STAINING
7. MICROBIAL DEGRADATION OF PLASTIC
• Surface Sterilization of Polystyrene.
• Media Preparation.
- 250ml of cooled Nutrient broth and potato dextrose broth was poured
into four 500ml conical flasks.
- The sterile pre-weighed plastic pieces were aseptically transformed
into respective medium.
- A loopful of Micrococcus Luteus a Masoniella was inoculated into
respective medium.
- These flasks were incubated at 37OC for 35, 45, and 55 days in shaking
incubators.
8. RECOVERY
• After 35 day of incubation, the removed
pieces from culture were shade diced and
weighed for final weight. The data were
recorded.
• The same procedure was also repeated for 45
and 55 days of incubation.
9. Determination of Degradation of
Plastics
• Measuring the weight lost by plastic after 35
of incubation.
% weight loss = Initial Weight – Final Weight
Initial weight x 100
14. Biodegradation
• The degradation of plastics by M.Luteus exposed at
35 days of incubation. Showed 19.6% of weight loss.
Similarity degradation of 45 and 55 days of incubation
showed 27.6%and 32 % respectively.
• The degradation of plastics by Masoniella sp
exposed at 35 days of incubation showed 8.8% of
weight loss. Similarly degradation of 45 and 55 days of
incubation showed 17.8% and 27.4% respectively.
17. CONCLUSION
• Micrococcus luteus and monosiella are very promising in
degrading the plastic.
• The bacteria caused the biodegration ranging from 19.6%
to 32% for plastics among the bacteria Micrococcus luteus
were found most active in degrade
38% of plastics.
• The fungi caused the biodegradation ranging from 8.8% to
27.4% for plastics. Among the fungi Masoniella sp were
found most active in degrading 27.4% of plastics.