Name: ALLIAH KATE LANGBIS Score:____________

Date Performed: 4/7/24 Date submitted: Section: NAA

ACTIVITY 7
MICROSCOPIC EXAMINATON OF BACTERIA

Danish Physician Hans Christian Gram (1853-1938) developed the Gram stain. When
freshly grown
bacterial cells are treated with the reagents used in the Gram stain, they can be categorized into one
of two classes: Gram (+) or Gram (-). Gram (+) class will appear blue or violets while the Gram (-)
class will appear red or pink.
The physiology and structure of a microorganism is intimately associated with its ability to
produce disease. The cell wall is a tough and rigid structure surrounding the cell. The rigid part is
the peptidoglycan. Peptidoglycans are the structural elements of almost all bacterial cell walls. They
constitute almost 95% of the cell wall in some Gram positive bacteria and as little as 5-10% of the
cell wall in Gram negative bacteria. The cell walls of gram positive bacteria have simpler chemical
nature than those of gram negative bacteria.
The bacterial cytoplasmic membrane is composed of a PHOSPHOLIPID LAYER and thus has
all of the general functions of a cell membrane such as acting as a permeability barrier for most
molecules and serving as the location for the transport of molecules into the cell.
The cytoplasm is the TOTAL OF EVERYTHING INSIDE of the cytoplasmic membrane. It
contains the Cytosol or the Amorphous Matrix to which nearly all other functions not conducted by
the cell membrane occur and it is where interior structures are suspended. It contains ribosomes,
storage granules, nucleoid, plasmids, and transposons. Other parts may include a capsule, flagella,
fimbriae, glycocalyx, and spores.
1. What is acid fast staining?
- Is a differential stain used to identify acid-fast organisms such as members of the genus
Mycobacterium. Acid-fast organisms are characterized by wax-like, nearly impermeable cell walls;
they contain mycolic acid and large amounts of fatty acids, waxes, and complex
lipids(www.austincc.edu)
- Acid-fast staining is indeed a differential staining technique that is utilized to identify acid-fast
organisms, which include members of the genus Mycobacterium. These organisms are
characterized by their wax-like, nearly impermeable cell walls that contain mycolic acid and large
amounts of fatty acids, waxes, and complex lipids. This staining technique is crucial in microbiology
for the identification of these specific types of bacteria, as their unique cell wall composition makes
them resistant to most common staining techniques.

2. Complete the table for Acid-fast staining

ZIEHL NEELSEN KINYOUN
Common name Hot stain Cold stain
Heating Yes (3-5 minutes) No
Primary stain Carbol Fuchsin Kinyoun Carbol Fuchsin (higher
concentration)
Decolorizer Acid-Alcohol (HCl-Ethanol) Acid-Alcohol (HCl-Ethanol)
Counterstain Methylene Blue/Malachite Green Methylene Blue/Malachite Green
3. Complete the tables
A. Eukaryotic cell functions.
Parts of the cell Function/s
Nuclear envelope Controls movement of materials between nucleus and cytoplasm
Chromatin DNA and protein complex containing genetic information
Nucleolus Site of ribosome production
Peroxisome Breaks down toxic molecules like hydrogen peroxide
Endoplasmic reticulum Network of membranes for protein synthesis (Rough ER) and
lipid/carbohydrate synthesis (Smooth ER)
Smooth E.R. Synthesis of lipids, carbohydrates, and detoxification
Microtubules Part of the cytoskeleton, provides structure and aids in cell movement and
division
Centrosome Organizes microtubules during cell division
Intermediate filaments Provide structural support and maintain cell shape
Microfilaments Involved in cell movement, muscle contraction, and cytokinesis (cell
division)
Plasma membrane Selectively permeable barrier controlling what enters and leaves the cell
Lysosomes Breakdown of waste, cellular debris, and foreign particles
Golgi apparatus Modifies, packages, and transports proteins and lipids
Cytoplasm Gel-like substance that suspends organelles and other cellular components
Mitochondria Powerhouse of the cell, produces energy (ATP) through cellular respiration

B. Prokaryotic Cell
Parts of the Function/s
cell
Plasmid Stores non-essential genes, often carrying genes for antibiotic resistance or toxins
Cell membrane Controls movement of materials in and out of the cell
Cell wall Provides structural support and protection against bursting
Capsule Protects the cell from phagocytosis (by white blood cells) and desiccation (drying out)
in some bacteria
Pili Facilitate attachment to other cells or surfaces
Flagella Enable movement (allow the cell to swim) / Enable movement (motility)

4. Complete the following:

A. Types of flagellum
Flagellum Description Example (name of bacteria)
Monotrichous Single flagellum at one end Pseudomonas aeruginosa
(polar)
Lophotrichous Cluster of flagella at one or both Vibrio cholerae
poles
Amphitrichous Single flagellum at both ends Salmonella typhi
(polar)
Peritrichous Numerous flagella distributed Escherichia coli
around the cell
5. Draw the following:
Coccus Diplococcus

Diplococcic encapsulated Staphylococci

Streptococci Sarcina
Tetrad Coccobacillus

Bacillus Diplobacillus

Palisades Streptobacilli
Vibrios Spirillum

Spirochetes Pleomorphic

Monotrichous Lophotrichous
 Peritrichous Amphitrichous

6. Give example of diseases and name of causative agent according to the following :
A. Diplococus – Streptococcus pneumonia
 Disease: Pneumonia, meningitis, sinusitis, and ear infections

B. Streptococcus – Streptococcus pyogenes (Group A Streptococcus)

 Disease: Strep throat, impetigo, scarlet fever, and rheumatic fever

C. Staphyloccous – Staphylococcus aureus

 Disease: Skin infections (impetigo, boils), pneumonia, food poisoning, and bloodstream
infections

D. Bacillus – Bacillus anthracis

pen_spark

Disease: Anthrax

E. Vibrio – Vibrio cholera

 Disease: Cholera

F. Spirillum – Treponema pallidum

 Disease: Syphilis

G. Spirochetes – Borrelia burgdorferi

 Disease: Lyme disease

H. Gram positive bacteria – Streptococcus pyogenes (Group A Streptococcus)

 Disease: Strep throat
 Other diseases caused by gram-positive bacteria: Pneumonia, impetigo,
endocarditis, osteomyelitis, and toxic shock syndrome.

I. Gram negative bacteria- Escherichia coli (E. coli)

 Disease: Urinary tract infection (UTI)
 Other diseases caused by gram-negative bacteria: Food poisoning, pneumonia,
meningitis, and sepsis.

J. Spore-forming bacteria – Clostridium tetani

 Disease: Tetanus
 Other diseases caused by spore-forming bacteria: Anthrax, botulism, and
pseudomembranous colitis.

7. What are the 10 most dangerous antibiotic-resistant bacteria? Illness caused? Antibiotic
resistance?
Virulence?

1. Methicillin-resistant Staphylococcus aureus (MRSA):

o Illness: MRSA can cause a range of illnesses, from skin infections and pneumonia to
bloodstream infections and sepsis.
o Antibiotic resistance: MRSA is resistant to all beta-lactam antibiotics, including penicillin
and methicillin.
o Virulence: MRSA can be highly virulent, especially in healthcare settings where people are
already weakened by illness.
2. Clostridioides difficile (C. Diff):
o Illness: C. Diff causes diarrhea, often after antibiotic use. It can also cause
pseudomembranous colitis, a severe inflammation of the inner lining of the colon.
o Antibiotic resistance: C. Diff is becoming increasingly resistant to commonly used
antibiotics, such as metronidazole and vancomycin.
o Virulence: C. Diff can be life-threatening, especially for older adults and people with
weakened immune systems.
3. Vancomycin-resistant Enterococci (VRE):
o Illness: VRE can cause a variety of infections, including urinary tract infections,
bloodstream infections, and pneumonia.
o Antibiotic resistance: VRE is resistant to vancomycin, one of the last-resort antibiotics for
treating serious infections.
o Virulence: VRE infections can be difficult to treat because of their antibiotic resistance.
4. Carbapenem-resistant Enterobacteriaceae (CRE):
o Illness: CRE can cause a variety of infections, including pneumonia, bloodstream infections,
and urinary tract infections.
o Antibiotic resistance: CRE are resistant to carbapenems, a class of broad-spectrum
antibiotics that are considered a last resort for treating many infections.
o Virulence: CRE infections can be difficult to treat because of their antibiotic resistance.
5. Drug-resistant Neisseria gonorrhoeae:
o Illness: Gonorrhea is a sexually transmitted infection (STI) that can cause a variety of
symptoms, including pelvic inflammatory disease in women and infertility in both men and
women.
o Antibiotic resistance: Strains of gonorrhea that are resistant to some of the antibiotics
used to treat it have emerged.
o Virulence: Untreated gonorrhea can lead to serious health complications.
6. Multidrug-resistant Pseudomonas aeruginosa:
o Illness: Pseudomonas aeruginosa can cause a variety of infections, including pneumonia,
bloodstream infections, and wound infections. It is a particular concern in healthcare
settings, where it can be difficult to eradicate.
o Antibiotic resistance: Pseudomonas aeruginosa is becoming increasingly resistant to
many antibiotics.
o Virulence: Pseudomonas aeruginosa can be a serious pathogen, especially for people with
weakened immune systems.
7. Drug-resistant Salmonella:
o Illness: Salmonella causes foodborne illness, which can cause symptoms such as diarrhea,
vomiting, and fever.
o Antibiotic resistance: Some strains of Salmonella are becoming resistant to commonly
used antibiotics.
o Virulence: Salmonella can cause serious illness, especially in young children and older
adults.
8. Drug-resistant Campylobacter:
o Illness: Campylobacter is a common cause of foodborne illness, which can cause symptoms
such as diarrhea, abdominal cramps, and fever.
o Antibiotic resistance: Some strains of Campylobacter are becoming resistant to
commonly used antibiotics.
o Virulence: Campylobacter can cause serious illness, especially in young children and older
adults.
9. Drug-resistant Shigella:
o Illness: Shigella causes shigellosis, a diarrheal illness that can be bloody.
o Antibiotic resistance: Some strains of Shigella are becoming resistant to commonly used
antibiotics.
o Virulence: Shigellosis can cause serious illness, especially in young children and older
adults.
10. Candida auris:
o Illness: Candida auris is a fungus that can cause serious infections, including bloodstream
infections and pneumonia.
o Antibiotic resistance: Candida auris is resistant to many antifungal drugs.
o Virulence: Candida auris infections can be difficult to treat because of their antifungal
resistance.

8.

Name Year First Illness caused Antibiotic Virulence (deadly,

documented resistance worrying,
(low, dangerous)
medium,
high)
Streptococcus 1884 Strep throat, Low Deadly (in rare,
pyogenes impetigo, scarlet severe cases)
fever, rheumatic
fever (complication)
Neisseria 1879 Gonorrhea (STD) Increasing, Worrying (can lead to
gonorrheae some strains infertility if
resistant untreated)
Mycobacterium Ancient Tuberculosis (TB) Medium Deadly
tuberculosis (evidence from (growing
9,000 year old concern)
remains)
Acinetobacter 1940s Hospital-acquired High Dangerous (often
baumannii infections difficult to treat due
(pneumonia, to resistance)
bloodstream)
Escherichia coli N/A (commonly Mostly harmless, Varies by Dangerous (some
 found in gut) some strains cause strain strains can cause
food poisoning, UTIs severe illness)
Klebsiella 1884 Pneumonia, Medium Dangerous (often
pneumoniae bloodstream (increasing hospital-acquired and
infections, UTIs concern) can be resistant to
antibiotics)
Clostridium 1938 Antibiotic-associated High Dangerous (can be
difficile diarrhea, life-threatening,
pseudomembranous especially for
colitis vulnerable
populations)
Pseudomonas 1882 Hospital-acquired High Dangerous (often
aeruginosa infections difficult to treat due
(pneumonia, to resistance)
wounds, burns)
Burkholderia 1950s Cystic fibrosis High Dangerous (especially
cepacia complications, lung for
infections immunocompromised
patients)
Staphylococcus 1961 Skin infections, High Deadly (especially in
aureus pneumonia, healthcare settings)
(MRSAmethicillin- bloodstream
resistant infections
Staphylococcus
aureus)

By: Longitude team. August 26, 2014. Longitude prize. http://longitudeprize.org/blog-post/10-

mostdangerous-antibiotic-resistant-bacteria