Pir Mehr Ali Shah

Arid Agriculture University, Rawalpindi

Office of the controller of Examinations
Final Exam / Spring 2021 (Paper Duration 12 hours)
To be filled by Teacher

Course No.: BCH 306 Course Title: General Microbiology (Theory)

Total Marks: 24 Date of Exam: 8-7-2021
Degree: BS (MIC) Semester: 2 nd Section: Morning * /Evening
Marks
Q.No. 1 2 3 4 5 6 7 8 9 10 Obtained/
Total Marks
Marks
Obtained
Total Marks in Words:
Name of the teacher: Sadia Mehmood Satti
Who taught the course:Signature of teacher / Examiner:

To be filled by Student

Registration No.: 20-ARID-2257 Name: Bisma Abid.

Answer the following questions.

Q.No.1. Explain the effectiveness of heat as a sterilizing agent? (3)

Answer:
Heat Sterilization.
Heat is mostly used and highly effective method for controlling the microbial growth.
In research and medical field, as well as to sterilizie food applying heat to bacterial
media is the most common method for the control of the bacterial growth. For
sterilization process, different techniques and tools are used.
Types Of Heat Sterilization.
Heat sterilization is of two types.
 Dry Heat
 Moist Heat
Dry Heat.
In dry heat sterilization, microbes are killed by charring. It causes oxidative damage
and denaturation of cellular proteins. Dry heat sterilization is carried out in hot air
oven. The temperature ranges from 160 to 180 degree Celsius for 2 hours.
Benefits.
 It provides good penetrability.
 It provides non-corrosiveness .
 It is used for sterilization of glass ware’s, surgical instruments and chemicals such as
glycerol, fats etc.
Moist heat.
Moist heat sterilization used hot water vapour as a sterilization agent. In it, three types
of temperate are used. It causes destruction of microorganisms by denaturation of
macro molecules, and proteins.
 Moist heat at temperature above 1000C
Autoclaving.
It is a common method for moist sterilization. It helps to kill bacteria , fungi and
spores but don't eliminate prions. In this, samples are placed in a srteam chamber. The
chamber is closed and heat up so that steam exhausts. Then pressure is applied so that
the temperature inside reaches to 121 degree.the temperature is set and maintained for
15 to 30 mints. The chamber is then allowed to cool or by passive heat dissipation.
 Moist heat at temperature below 1000C
Pasteurization.
The process is used for sterilization of food like fruits, vegetable juices and milk etc.
Water Batch.
For sterilization of vaccines, water bath is used. For sterilization of body fluids and
serum samples at 160 degrees Celsius for 60 minutes.
 Moist heat at temperature 1000C.
Boiling.
Boiling of samples in water for 15 minutes kills the microorganisms except some
fungal spores.

Q.No.2. How does antimicrobial agents work. (3)

Answer:
Antimicrobial Agents.
An antimicrobial agent is a natural or synthetic substance that kills or stop the growth of the
microorganisms such as fungi, algae and bacteria. They are used to destroy microorganisms
to prevent them for developing.
Example.
Penicillin G, penicillin V and syphills etc.
Working.
They work at cellular level to destroy and to prevent the growth of microorganisms. An
inhospitable environment is created for microorganisms. Antimicrobial protect products like
toys and textiles etc.
Agents used.
The agents that are used includes.
 Disinfectants
 Antiseptic.
 Sanitizer.
 Cidal and static agent.
Disinfectants.
It is an agent that kills or destroys bacteria. It inactivates or destroy microorganism on inert
surface. It is used in hospitals, kitchens and clinics etc.
Antiseptic.
It kills or inhibits growth of the microbes but it is safe to used on human tissues. It is used to
reduce infections and sepsis etc.
Sanitizer.
Sanitizer is used to reduce microbial numbers to a safe level. It is used to kill viruses and
bacteria etc.
Cidal And Static Agent.
It kills microorganism and viruses. They temporarily reduce the growth of
microorganisms.static agent inhibit the growth.

Q.No.3. Justify the protein synthesis inhibition using antibiotics? (3)

Answer:
Protein synthesis Inhibition.
Antibiotics inhibits the protein synthesis inhibition by binding to the 30S subunit of bacterial
ribosome. Aminoglycosides are protein synthesis inhibitors that acts by impairing bacterial
protein synthesis through binding to prokaryotic ribosomes.
Protein synthesis is an important process necessary for both human cells and bacterial cells
multiplication and survival. Enzymes and other cellular structures are made of proteins.
Antibiotics should be selected very carefully so that they not interfere with protein synthesis
in human cells but to do in bacterial cell. Humans have 80S ribosomes whereas bacterium
have 70S ribosome. These antibacterial agent target bacterial protein synthesis by binding to
the 30S or 50S subunits of the intracellular ribosomes. This activity does nothing for the 80S
ribosomes in human but results in disruption of normal cellular metabolism of bacteria.
Example.
Chloramphenicol is responsible to block the peptidyl transfer step of elongation on 50S
ribosomal subunit in both bacteria and mitochondria.

No.4. Explain the mode of action of beta-lactams. (3)

Answer:

Beta-Lactams.

They are bactericidal and act by inhibiting the synthesis of peptidoglycan layer of bacterial
cell walls. The beta-lactam ring stands alone and not fused to another ring.  They work by
inactivating serine beta-lactamases,the enzymes that hydrolyze and inactivate the beta-
lactam ring .

Mode of Action.

Antibiotic inhibitors of cell wall synthesis block the production of  peptidoglycan.The cell
wall is important for normal functioning of the bacterial cell. Cross-linking between
peptidoglycan chains forms a strong, mesh-like structure that gives the cell wall structure
rigidity, and protects the underlying cell membrane from osmotic damage when water
moving into the cell by osmosis could cause it to burst, or lyse. Disruption of the
peptidoglycan layer of the cell wall can result in cell lysis.

The structural similarity between B lactam antibiotics and D alanyl facilitate their binding to
active of penicillin binding protein PBPS.

Example.

ß-lactam antibiotics contain a core chemical structure called a ß-lactam ring which
determines the mode of action of this class of antibiotics.The ß-lactam antibiotics
interfere with the formation of the peptidoglycan cross-links, thereby weakening the cell
wall.These drugs contain Beta lactam rings in their structure. These all are bactericidal in
nature .

BETA LACTAM COMPOUNDS :

 Penicillin.
Mode of action :Prevents bacterial cell wall synthesis by binding to and inhibiting cell
wall transpeptidases.
 Cephalosporin.
Mode of action :Prevents bacterial cell wall synthesis by binding to and inhibiting cell
wall transpeptidases.
 Monobactams.
Mode of action :Prevents bacterial cell wall synthesis by binding to and inhibiting cell
wall transpeptidases
 Carbapenems
Mode of action :Prevents bacterial cell wall synthesis by binding to and inhibiting cell
wall transpeptidases
Q. No.5. State the antibiotic resistance mechanisms. (3
Answer:

Antibiotic resistance.
Antibiotic resistance occurs when the germs are not responding to the antibiotics that are
designed to kill them. It is the phenomenon that susceptibility of pathogenic microorganisms
to antibiotic becomes so low and even loses after microorganism comes in contact with
antibiotic a lot of time.
Cross antibiotic resistance.
Cross antibiotic resistance occurs between antibiotic of the same family, to which bacteria
may not have been exposed. Bacteria show resistance to one antibiotic, they are also resistant
to some other antibiotics.
Mechanism of Antibiotic resistance.
Enzyme inactivation of the antibiotic.
The first ever resistance to developed is resistance to penicillin that is beta-lactam
antibiotics that can be inactivated by beta-lactamases. They are family of enzymes
that are found in many bacterial pathogens.
Alteration of the target side of the Antibiotic.
It is a common mechanism that is used by bacteria to become resistant to antibiotic by
modifying the target of the antibiotic.methicillin resistances among staphylococcus
aureus.
Active transport of the antibiotic out of the bacterial cell.
Bacteria will be resistant to its action if an antibiotic cannot reach its target. Actively
transporting antibiotics out of the cell decreases their concentration inside the cell, so
that they cannot build up to a high enough concentration to exert the effect on their
target.
Example. Tetracycline etc.
Decreased Permeability of bacterial cell wall to antibiotic.
Alteration in porin proteins that forms channel in the cell membrane. Example :
resistance of Pseudomonas aeruginosa to a variety of penicillin and Cephalosporin.

Q.No.6. How would you argue transformation is different from generalized

transduction? (3)
Answer:
Transformation. Generalized transduction.
Transformation is a transfer of genetic In generalized transduction defective virus
material from one cell to another and can particle incorporate fragments of host DNA
change the genetic makeup of recipient cell. to other recipient cell.

In transformation a non-reciprocal type of Generalized transduction may occur via

recombination takes place. recombination.
It happens as a time limited response to It happens when a phage is in the lytic stage,
environmental conditions like cell density the moment when the viral DNA is packaged
and starvation. into phage heads.

Q.No.7. How specialized transduction is different from generalized transduction? (3)

Specialized transduction. Generalized transduction.
Specialized transduction is the process by In generalized transduction defective virus
which a restricted set of bacterial genes is particle incorporate fragments of host DNA
to other recipient cell.
transferred to another bacterium. 

Specialized transduction is done by Generalized transduction is done by virulent

temperate bacteriophages in which bacterial bacteriophages in which bacterial cell is lysed
cell is not lysed. when new bacteriophages are released.
Specialized transducing phages carry only Generalized transducing phages can carry
restricted parts of the bacterial chromosome any part of the chromosome
Certain donor gene. Any donor gene.
Temperate phage. Virulent phage.

R. No.8. State how striking features of Archaea making them different from bacteria.
(3)

Archaea are tiny organisms, less than one micron long. When observed under high-power
light microscope, they still look like dots. Archaeal physical features can be distinguished by
electron microscope. Similar to bacteria, archaea have no internal membranes and their
genome exists as single loop called plasmid. Several archaea have outer membrane separating
cell from environment. Within cytoplasm Archaeal cellular functions are going on. All
archaea have cell wall, semi-rigid maintaining cell shape and chemical equilibrium.

All structural features of archaea may have similarities with other bacteria or eukaryotes but
chemical composition is very different. Like the cell wall in bacteria is composed of
peptidoglycan while archaea do not have this cell wall composition.

Striking Features.
The absence or presence of peptidoglycan is a distinguishing feature between the archaea and
bacteria. Different type of cell walls are present in archaea.
Bacteria. Archaea.
Bacteria contain peptidoglycan in their cell Archaea lack peptidoglycan.
wall.
The cell membrane in bacteria is a lipid In archaea, it can be a mono-layer.
bilayer.
Bacteria contains fatty acids. Archaea contains phytanyl.

Bacteria are divided into gram positive and Archaea are divided into different groups like
gram negative depending on gram staining methanogens, thermophiles etc
Example. Example.
Staphylococcus aureus. Staphylothermus marinus
Histones are absent. Histones are present.