Brucal, Mei Bhell P.

BSEd Science 3
Activity 2
Cell and Molecular Biology

Answer the following:

1. Compare and contrast the following using standard bases:
a. Plant and animal cells:

Animal Cell

Plant cell

Cell wall Absent Present (formed of cellulose)

Shape Round (irregular shape) Rectangular (fixed shape)

Vacuole One or more small One, large central vacuole taking up

vacuoles (much smaller to 90% of cell volume.
than plant cells).

Centrioles Present in all animal Only present in lower plant forms

cells (e.g. chlamydomonas)

Chloroplast Absent Plant cells have chloroplasts to

make their own food.

Cytoplasm Present Present

Ribosomes Present Present

Mitochondria Present Present

Plastids Absent Present

Endoplasmic Present Present

Reticulum (Smooth
and Rough)

Peroxisomes Present Present

Golgi Apparatus Present Present

Plasma Membrane Only cell membrane Cell wall and a cell membrane

Microtubules/ Present Present

Microfilaments

Flagella Present in some cells Present in some cells (e.g. sperm of

( e.g. mammalian sperm bryophytes and pteridophytes,
cells) cycads and Ginkgo)

Lysosomes Lysosomes occur in Lysosomes usually not evident.

cytoplasm.

Nucleus Present Present

Cilia Present Most plant cells do not contain cilia.

b. Prokaryotic and eukaryotic cells:

Similarities:

1. Cell Membrane:Both have a lipid bilayer that acts as a barrier.

2. Genetic Material:Both use DNA as the basis for their genetic information.

3. Ribosomes:Both have ribosomes which help in protein synthesis.

4. Cytoplasm:Both have cytoplasm, where biochemical reactions take place.

Differences:

1. Cell Size:Eukaryotic cells are generally larger than prokaryotic cells.

2. Cell Arrangement:Eukaryotes can be multicellular, while prokaryotes are usually

unicellular.

3. Nucleus:Eukaryotic cells have a true nucleus, while prokaryotic cells have a nucleoid.

4. DNA Structure:Eukaryotic DNA is linear and organized into chromosomes, while

prokaryotic DNA is circular.
5. Organelles:Eukaryotic cells have membrane-bound organelles, while prokaryotic cells
do not.

6. Ribosome Size:Eukaryotic ribosomes are larger than prokaryotic ribosomes.

7.Cytoskeleton: Only eukaryotic cells have a complex cytoskeleton.

8. Reproduction:Most eukaryotes reproduce sexually, while prokaryotes reproduce

asexually.

9. Cell Division:Eukaryotic cells divide by mitosis, while prokaryotic cells divide by binary
fission.

c. Gram-positive and Gram-negative bacteria:

Gram positive bacteria Gram negative bacteria

Distinctive purple appearance after gram Pale reddish color after gram staining
staining

Bacteria include all staphylococci, all Bacteria include enterobacter species,

streptococci and some listeria species salmonella species and pseudomonas
species

Thick peptidoglycan layer Thin peptidoglycan layer

No outer lipid membrane Outer lipid membrane present

No O-specific side chains present O-specific side chains present

Teichoic and lipoteichoic acids present Teichoic and lipoteichoic acids not present

2. Draw the structure of Gram + and Gram – bacterial cell wall. Label the parts.
a. Differentiate as to (a) shape, (b)appendages, (c) diseases caused by them and (d)
antimicrobial resistance;
a. Shape:
- Gram-positive bacteria
Have various shapes, including cocci (spherical), bacilli (rod-shaped), and spirilla
(spiral-shaped). Examples include Staphylococcus and Streptococcus.
- Gram-negative bacteria
Also exhibit various shapes, such as cocci, bacilli, and spirilla. Examples include
Escherichia coli and Pseudomonas aeruginosa.
b. Appendages:
- Gram-positive bacteria
Generally have fewer appendages compared to Gram-negative bacteria. They
possess flagella for movement and pili for attachment.
- Gram-negative bacteria
Have more complex appendages. They have flagella for movement, pili for
attachment, and an outer membrane that contains lipopolysaccharides (LPS).
c. Diseases caused by them:
- Gram-positive bacteria:
Responsible for various diseases. For example, Staphylococcus aureus can
cause skin infections, pneumonia, and bloodstream infections. Streptococcus
pneumoniae is known for causing pneumonia, meningitis, and ear infections.
Clostridium difficile is associated with antibiotic-associated diarrhea.
- Gram-negative bacteria:
Also implicated in a wide range of diseases. Escherichia coli can cause urinary
tract infections, gastrointestinal infections, and bloodstream infections. Salmonella
species are responsible for foodborne illnesses. Neisseria meningitidis causes
meningitis.
d. Antimicrobial resistance:
- Gram-positive bacteria:
Shown varying degrees of antimicrobial resistance. Methicillin-resistant
Staphylococcus aureus (MRSA) is a well-known example of resistance in Gram-positive
bacteria. Some strains of Streptococcus pneumoniae have also developed resistance to
antibiotics.
- Gram-negative bacteria:
Known for their higher propensity for antimicrobial resistance. They possess
mechanisms such as efflux pumps and enzymes that can degrade antibiotics. Examples
include carbapenem-resistant Enterobacteriaceae (CRE) and multidrug-resistant
Pseudomonas aeruginosa.

b. Give examples of Gram + and Gram – bacteria and their specific staining
techniques.
Gram positive bacteria are surrounded by a single thick peptidoglycan cell wall
and are therefore termed monoderms.On the other hand, gram negative bacteria have a
much thinner peptidoglycan cell wall, but in addition they have an outer membrane
containing lipopolysaccharides surrounding the cell and are consequently termed
diderms.

B. Gram Positive Bacteria:

1.Actinomyces:
 Actinomyces can be stained using the Gram staining technique, where crystal
violet is applied, followed by iodine, decolorization with alcohol or acetone, and a
counterstain such as safranin.

3. Bacillus:

Bacillus can also be stained using the Gram staining technique, which involves
crystal violet, iodine, decolorization, and a counterstain.

4. Clostridium:

Clostridium can be stained using the Gram staining technique as well. The steps
include crystal violet, iodine, decolorization, and a counterstain.

Gram Negative Bacteria:

1. Escherichia coli (E. coli):

E. coli can be stained using the Gram staining technique. It involves crystal
violet, iodine, decolorization, and a counterstain.

2. Salmonella:

Salmonella can also be stained using the Gram staining technique. The steps
include crystal violet, iodine, decolorization, and a counterstain.

2. Shigella:

Shigella is typically stained using the Gram staining technique. Crystal violet,
iodine, decolorization, and a counterstain are applied.

3. What are the different steps in counting cells using hemocytometer?

The following are the steps using hemocytometer:

1. Bring adherent and semi-adherent cells into suspension using trypsin/EDTA. Make
sure to resuspend them in a fresh medium with a volume at least equal to the trypsin
volume. If the cells grow in clumps, centrifuge and resuspend them in a small volume,
then gently break up the clumps by pipetting.

2. Under sterile conditions, take 100-200 μL of the cell suspension.

3. Add an equal volume of Trypan Blue (dilution factor = 2) and mix gently by pipetting.

4. Clean the hemocytometer.

5. Moisten the coverslip with water or breath, then slide it over the chamber using slight
pressure until Newton's refraction rings appear (rainbow-like rings).

6. Fill both sides of the chamber with approximately 5-10 μL of cell suspension.

7. View the chamber under an inverted phase contrast microscope with 20x
magnification.

8. Count the number of viable cells (bright cells) and non-viable cells (stained blue). It is
recommended to count more than 100 cells to improve accuracy. Take note of the
number of squares counted to reach over 100 cells.

9. Use the provided equations to calculate the concentration of viable and non-viable
cells, as well as the percentage of viable cells.

4.What are the best things to do if there are too many cells in the hemocytometer
chamber thatone will be counting?

Many researchers have studied how to handle too many cells in a

hemocytometer chamber. One common method is to dilute the cell sample by mixing it
with a buffer or culture medium. This reduces the cell concentration. Another option is to
count fewer squares in the grid and then estimate the total cell count. If these methods
don't work, using a different counting chamber designed for high cell concentrations
could be helpful. Further,increasing the magnification can also make it easier to see
individual cells. To ensure accurate results, it's recommended to count multiple times
and calculate the average. If available, automated cell counting instruments can be fast
and accurate. Consistent practice and a methodical approach are the key to successful
cell counting. It is said that it is important to take enough time and not rush the process.

References:
 https://www.diffen.com/difference/Animal_Cell_vs_Plant_Cell#:~:text=Animal%20cells
%20are%20mostly%20round,nucleus%2C%20mitochondria%20and%20endoplasmic
%20reticulum.

 https://www.news-medical.net/life-sciences/Eukaryotic-and-Prokaryotic-Cells-
Similarities-and-Differences.aspx

 Gram Positive vs Gram Negative | Technology Networks

 https://microbiologyinfo.com/gram-staining-principle-procedure-interpretation-examples-
and-animation/

 https://www.ncbi.nlm.nih.gov/books/NBK562156/
 https://microbeonline.com/gram-staining-principle-procedure-results/

 https://openstax.org/books/microbiology/pages/2-4-staining-microscopic-specimens

 https://www.sigmaaldrich.com/PH/en

 https://chemometec.com/how-to-count-cells-with-a-hemocytometer/