MICRO BIO LAB FINAL

Clostridium botulinum
Clostridium tetani
bacillus anthracis

Bacillus subtilis

Prepare smear
place paper towel over the slide
flood with malachite green and heat with flame-- stains the endospore (5-10mins)
rinse well with water (decolorization method)
flood slide for one minute wih safranin to stain the vegetative cell

Endospore stain - b. subtilis

Bacillus
clostridium

Trypticase Soy Agar
testing for general purpose
complex
not selective
not differential

Plate Count Agar
testing for general purpose
complex
not selective
not differential

Testing for amylase activity
complex
not selective
differential:add iodine to detect a clearing of starch around the growth

Starch agar
left side indicates the absence of intact starch which tests positive for the reaction of amylase

right side indicates that it was unable to break down starch, so it tests negative reaction for this test.

Testing for caseinase activity
complex
not selective
differential: clearing of skim milk from around growth indicates a positive reaction

Hydrolysis of the milk protein casein, by the exoenzyme caseinase

negative= negative for caseinase production (not caseinolytic)

positive= positive for caseinase (caseinolytic)

Testing for ammonification
complex
not selective
differential: add Nessler's to detect te production of ammonia

Testing for denitrification
complex
not selective
differential: Bubbles in durham vial indicate nitrogen gas production. add sulfamic acid to detect nitrite production from nitrate. Add zinc to confirm the presence of nitrate.

Testing for coliforms
complex
selective: sodium lauryl sulfate selects for coliforms
differential: Lactose is the fermentable sugar. Gas production in Durham vial. Beta-D- Glucuronidase cleaves MUG to produce a product which fluoresces under UV light

LST - MUG test