WORKSHEET 2 gram positive purple

DETERMINATION OF MICROBIAL gram negative pink

MOTILITY
2 Escherichia coli
(a) Microoganisms in pond true Shape rods / bacilli
motility Arrangement single
(b) Saccharomyces cerevisae Gram reaction Gram (-), pink
Brownian movement
2 Pseudomonas aeruginosa
Brownian movement molecules Shape Rod
suspending liquid colliding with organisms, Arrangement Single
irregular and vibrating Gram rxn Gram (-), pink
True motility consisteth path / maybe
twisting or turning 3 Staphylococcus aureus
Shape cocci
1 Differentiate the motility of Arrangement clusters
microorganisms obsered in wet mount and Gram rxn Gram (+), purpel
hanging drop (movmeent)
Wet mount -> exerts pressure on the 4 Bacillus subtilis
microorganism, movement in hindered Shape Rod
Hanging drop -> having space would allow Arrangement chains
it to move freely and doesnt affect its Gram rxn Gram (+), purple
ability to move
STAINING
SKETCH Crystal Violet Primary Stain (30
Micrococcus luteus sec)
Proteus vulgaris (+ motile) o + and - Purple
Staphylococcus aureus
Gram Iodine Mandant, binds and
traps crystal (1 min) violet
Saccharomyces cerevisae
o + and - Purple
WORKSHEET 3 Ethanol decolorization,
MICROSCOPIC EXAMINARION OF dehydrates and decolors (15 sec)
STAINED BACTERIA o + Purple
o - Colorless
Teeth scraping (positive staining) Safranin counterstain (30 sec)
Pseudomonas aeruginosa (negative o + Purple
staining) o - Pink or Red

1 Difference between positive staining and Why is it necessary to use a young culture
negative staining based on the structure in gram staining? (24 hour old culture)
extrernal to the cell wall
Gram positive bacteria one membrane is - Young culture have the best form of
surrounded by a thick layer of CW (made of peptidoglycxan in their cell wall. CW
peptidogylcan) becomes heavy as it ages making the gram
Gram negative bacteria inner layer test no longer accurate.
followed by thin layer or peptidoglycan and
outer membrance
difference in thickness of cell wall
gives difference in color