W O R K S H E E T

Activity No. 8

Name/s : Jingco, Morones, Daham, Magnaye, Date Performed:

Dichuasido Date Submitted :

Group No. : 1

Observation:

What color would the bacterial cells have when viewed microscopically after
the following treatment?

Gram Ammonium Gram’s iodine Ethyl Alcohol Safranin

reaction oxalate
Crystal violet
Gram positive purple Purple Purple purple

Gram purple purple colorless Pink/red

negative

Questions:
1. Of what value is gram staining?
In microbiology, the Gram stain is the most effective staining technique. It is used to
distinguish between gram positive and gram negative cells. As a result, it is a differential
dye. Differences in cell walls differentiate gram negative and gram positive cells.

2. Why are the steps in gram-staining so carefully standardized?

These are (1) their widespread presence in soil, (2) their resistance to heat in
traditional industrial processes such as pasteurization, (3) the adhesive properties of
specific spores that aid in their attachment to processing equipment, and (4) their ability
to germinate and expand in favorable conditions.

1. Why is it necessary to employ a spore stain rather than rely on a diagnosis of

sporing from examination of gram stain?
Endospore formation is an essential feature of certain bacteria, allowing them to
withstand unfavorable environmental conditions such as desiccation, chemical exposure,
excessive heat, and so on. Acterial endospores are the most immune structures of all
living species, and they can remain dormant and dehydrated for hundreds of years (even
some documented at thousands of years). Endospores are not capable of reproduction:
Within the vegetative cell, one spore develops. One vegetative cell will be formed when
the spore germinates.
 2. Of what importance are spore-forming bacteria in the food industry?
Bacterial spores are a source of concern for the food industry because of their
ability to survive processing, as well as the various measures designed to
destroy vegetative cells, and their ability to germinate and expand in food,
lowering its protection and shelf life.

3. Of what practical significance are capsule forming bacteria in industry and

medicine?
The capsule is referred to as a virulence factor because it increases bacteria's
capacity to induce disease (e.g. prevents phagocytosis). The capsule will shield cells
from eukaryotic cells like macrophages engulfing them. Phagocytosis can involve the
presence of a capsule-specific antibody.

4. Why is mordant necessary for staining of bacterial flagella?

Since bacterial flagella are exceptionally slender and delicate a unique stain
(flagella stain) is readied that contains a stringent. This stringent permits heaping of the
stain on the flagella, expanding the thickness until they become noticeable. Different
plans of flagella are seen on various cells

5. Differentiate differential stain from simple stain.

Simple stain uses 1 dye to increase contrast of cells. A simple stain determines size,
shape, and arrangement of cells but cannot differentiate between types of bacteria. A
differential stain uses 2 or more dyes to differentiate between organisms or between cell
structures. A differential stain is a specific type of staining that allows for microbe
identification and distinguishing between cells in a mixed sample. This is very different
from simple staining techniques that simply add color and contrast to a slide. Simple
staining involves adding a basic, cationic dye to the organism. The positive dye is
attracted to the negative cell wall and cytoplasm, resulting in stained cells. A simple
stain reacts with all microbes in the same way. This is helpful in defining cell
arrangement, size and morphology. This is much different from differential stains. There
are several different kinds of differential staining dyes that can obtain different staining
results, and they can be used in a serial manner. Each dye reacts with a particular
physical property of a cell.

VI. Conclusion
In conclusion, in preparing a smear bacterial cells, we must follow these steps: (1)
Place one needle of solid bacterial growth or two loops. of liquid bacterial growth in
the center of a clean slide.
(2) If working from a solid medium, add one drop (and only one drop)
(3) Now, with your inoculating loop, mix the specimen with the water
(4) Place the slide on a slide warmer and wait for it to dry. Differential staining is a
staining technique that employs the use of several chemical stains. Multiple stains
may help distinguish between various microorganisms or different structures/cellular
components within a single organism. Lastly, in classifying gram positive-negative,
gram-positive bacteria look purple-blue under a microscope because their dense
peptidoglycan membrane will store the pigment. Because of the positive outcome, the
bacteria is known as gram-positive bacteria. Gram-bad bacteria produce a pink-red
dye. Since their peptidoglycan coating is thinner, the blue colour is lost. The outcome
of the test is negative.