`NAME: JAYSETTE JUNE N.

SACUPASO RATING: ________________________

GROUP NO. 2 DATE: 03/13/21

ACTIVITY NO. 6

GRAM STAINING

While working in a laboratory in the morgue of a Berlin hospital in the 1880s, a Danish
physician named Hans Christian Gram developed what was to become the most
important of all bacterial staining procedures. He was developing a staining technique
that would enable him to see bacteria in the lung tissues of patients who had died of
pneumonia. The procedure he developed—now called the Gram stain—demonstrated
that two general categories of bacteria cause pneumonia: some of them stained blue
and some of them stained red. The blue ones came to be known as Gram-positive
bacteria, and the red ones came to be known as Gram-negative bacteria. It was not
until 1963 that the mechanism of Gram differentiation was explained by M.R.J. Salton.

OBJECTIVES:
At the end of the activity the student would be able to
1. learn the technique in gram staining procedure
2. identify and classify bacteria according to their reaction to gram stain.
3. Know the other methods of bacterial staining.

MATERIALS: Microbiology book and e-book

PROCEDURES: Answer the following research questions. Write legibly.

RESEARCH QUESTIONS:

1. Draw and label the gram staining technique.

Gram Staining Method
 2. Illustrate and label the cell wall of gram negative and gram positive bacteria

Cell wall of Gram Positive Bacteria

Cell Wall of Gram Negative Bacteria

RESEARCH QUESTIONS
3. Discuss the principle of gram staining technique.
- The gram stain technique is based on the differential structure of the cellular
membranes and cell walls of the two groups.
- The cells appear colorless. To make the colorless cells visible, a secondary
stain, safranin, is applied, leaving the gram-negative cells pink.

4. Give similarities of gram positive and gram-negative microorganism.

 - Both lack membrane bound organelles. Both are prokaryotic in nature. In both
groups, cell wall is made up of peptidoglycan
- Gram positive ang gram negative bacteria undergo genetic recombination
through transduction, transformation and conjugation.

5. Tabulate the difference between gram positive and gram-negative

microorganisms.

Gram positive bacteria Gram negative bacteria

Simple cell wall
Thich peptidoglycan cell wall layer
No outer lipopolysaccharide wall layer
Retain crystal violet/iodine
Appear (blue/purple)
More complex cell wall
Thin peptidoglycan cell wall layer
Outer lipopolysaccharide wall layer
Retain safranin
Appear (pink/red)

6. Give examples of common gram positive and gram-negative microorganism and

the disease/s they cause.

Gram negative bacteria and infections and they cause

- (E. coli)- food poisoning urinary tract infections, gastroenteritis and new born
meningitis
- Pseudomonas aeruginosa- lung and urinary tract infections
- Klebsiella- meningitis, and lung, urinary tract, and bloodstream infections
- Acinetobacter baumannii- several types of infections in wounded soldiers
- Neisseria gonorrhea- a sexually transmitted disease
- Enterobacteriaceae- urinary tract, lung and bloodstream infections, and food
poisoning

Gram positive and infections and they cause

- Mycobacterium tuberculosis- cause tuberculosis
- M. leprae, - cause leprosy
- Corynebacterium- cause diphtheria

REFERENCES:
www.gramstain.ppt
www.courses.lumenlearning.com
www.winchesterhospital.com
www.byjus.com