NEED OF ASSIGNMENT:

Composting is the biological conversion of organic wastes to a stabilized product that can be used as soil amendment. Different types of MOS present in agricultural compost. This assignment basically done for the learning of isolation technique of MOS.

AIM:

Isolation of bacteria from Agriculture compost.

REQUIREMENT:

  1. Agricultural compost
  2. Petri plates
  3. Conical flasks
  4. Spreader
  5. Test tubes
  6. Autoclave
  7. Incubator
  8. LAF
  9. Microtips and micropipete.

Protocol:

  1. Prepare PDA and NA media .
  2. Autoclaved media,petri plates,distilled water,test tubes,spreader,microtips at 121C temp and 15 lbp pressure for 20 min.
  3. For Maintaining asceptic condition : At the same time, clean LAF(laminar air flow) using aceton /80% ethanol and ON UV light for 15-20 min.after 20 min OFF UV light and and ON blower and day light.
Image 1: Autoclave.
  1. Then all remaining procedure carried out under asceptic contion
  2. After sterilization, media poured into petri plates (approximately 30 ml/plate)
Image 2: Pouring of media into petri plates.
  1. Serial dilution: for serial dilution ,weigh 1 gm of trichoderma powder.Take 10 sterile test tube and filled it with 9 ml distilled water each.Dissolve 1 gm Agricultural compost in it and subsequently transfer 1 ml aliquot from test tube 1 to last.Then finally we have concentration of trichoderma would be 10-1,10-2,…..10-9
  2. After solidification of media, add 0.2 ml of aliquot from test tube (with concentration 10-4 ) on petri plate.And spread equally on whole petri plate.
  3. Follow same protocol for all the test tubes with different concentration and innoculate on the different petri plate.
  4. Packed petri plates with parafilm. And kept it in incubator at 37 C temp for 3/4 days.
Image 3 & 4 : Microbial growth on media plates.