**4.5 Gram staining**

A single colony from culture plate of isolated degrading bacteria was picked using sterile inoculation loop and mixed with a few drops of distilled water to form a smear on microscopic slide. Then, the smear was air dried and heat fixed by passing through the flame on bunsen burner for a few seconds. Next the primary stain, crystal violet was applied on the smear for 1 minute and then rinsed with water. Then the smear was covered with iodine for one minute before rinsing with water and ethyl alcohol was used to decolorize the slide for.

30 seconds. After decolorizing, safranin was added for 30 seconds before rinsing with water. Then the slides were observed under light microscope to identify and distinguish between gram positive and gram-negative bacteria.
