Pir Mehr Ali Shah

Arid Agriculture University, Rawalpindi

Office of the controller of Examinations
Mid Exam / Spring 2020 (Paper Duration 48 hours)
To be filled by Teacher

Course No.: MICRO-102 Course title: General Veterinary Microbiology

Total Marks: 12 Date of Exam: 19-06-2020
Degree: DVM Semester: 2nd Section: Morning/Evening
Marks
Q. No. 1 2 3 4 5 6 7 8 9 10 Obtained/
TotalMarks
Marks
Obtained

Total Marks in Words:

Name of the teacher: Dr. Saif-ur-Rehman/Dr. Zahid Manzoor

Who taught the course: Signature of teacher / Examiner:

To be filled by Student

Registration No.: 19-ARID-2248 Name: Sidra Zamir

Answer the following questions.

Q. No. 1. Describe bacterial structures that might be used as antibiotic targets & explain why?
(Marks 04)
Answer: The structures of bacteria which act as targets for antibiotics may include their
1) Cell walls and cell membranes
Peptidoglycan is the major component of the cell wall of bacteria which is made up of
disaccharides (NAM and NAG) and amino acids. These structures are involved in the
maintenance of shape of cell and its integrity. Also cell wall is a barrier between the outside
and inside environment which allows only certain molecules to pass through it. Thus, when
antibiotics (such as penicillin’s) attacks on the cell wall, it interferes with the synthesis of
peptidoglycan and the bacteria loses its cellular integrity with increasing pressure of outside
fluid on the bacteria. Also the cell wall of bacteria is involved in binary fission, so when the
cell wall is destroyed bacteria will not be able to divide thus, causing its death. Plasma
membrane of bacteria is made up of phospholipids and proteins and is important for the
bacterial cell because it allow only selective molecules to pass through it thereby blocking
large molecules such as proteins. Thus, antibiotics attacks plasma membrane thereby
affecting its permeability. Loss of plasma membrane will cause any chemical to enter into
 the bacteria as there will be no barrier left between the interior of bacteria and the
extracellular fluid thus, causing its death.
2) Nucleic acids (DNA AND RNA)
The DNA of bacteria is made up of a single strand of chromosome which is arranged in
circular pattern with associated proteins and RNA called as bacterial chromosome. The
chromosome contains the DNA made up of genes. The genes carries the genetic
information. So antibiotics attack the DNA to cease all the vital functions occurring within
the cell and to inhibit the production of RNA from DNA (transcription). Thereby, affecting the
expression of gene.
3) Ribosomes
A ribosome (made up of ribonucleoproteins) is actually the site where proteins are
synthesized they are present in two forms within the cell that is either attached to the
membrane of cytoplasm or present freely within the cytoplasm. A ribosome is involved
in the synthesis of proteins from RNA. Antibiotics target ribosomes because when no
proteins will be formed in the body then all the cell processes will stop as proteins are
integral for structure and many biological processes. Thus, the RNA will not get
translated into proteins and protein synthesis will be inhibited.
Thus, antibiotics acts on these sites because these are the major sites on which the survival of
bacteria depends. Without any of these structures bacteria will die. So they target the core areas of
bacteria.
Q. No. 2. What type of conditions & chemicals will you provide to grow bacteria in the laboratory?
(Marks 04)
Answer: In order to grow microorganisms in vitro, a number of its requirements (physical and
chemical) must be fulfilled.
Physical requirements: It includes,
1) Temperature
Optimum temperature (37 degrees) is best for the growth of microorganisms because at this
temperature the microbes grows best. Depending upon microorganisms, their temperature
preference varies from type to type. Such as,
Psychrophilic (Which requires cold temperatures for growth such as 0-20 degrees)
Mesophilic (Which requires moderate temperature for growth such as 25-40 degrees)
Thermophilic (Which requires high temperatures for growth such as50-60 degrees)
Archaea bacteria (requires very high temperature of 80 degrees for growth)

2) pH
Optimum pH is best for the growth of microorganisms as it shows its maximum activity at
this pH. Different microbes require different pH for its growth such as,
 Acidophilic (these microbes requires an acidic pH for their growth)
Neutrophilic (these microbes requires neutral pH of 7 for their growth)
Alkaphiles (these microbes requires a low pH that is, lightly basic pH for their growth)

3) Osmotic Pressure
Hypertonic environments (containing large amount of dissolved substances) slows down the
growth of bacteria this is because when the environment contains large amounts of
dissolved substance than water then it will cause the water from the cells to move out in the
environment thereby shrinking the cytoplasm, a process termed as plasmolysis.
Halophiles (such microbes require salt concentration to be high in order to work)
Obligate halophiles (Salt concentration in the environment is necessary for them)
Facultative halophiles (Salt concentration is not necessary for them but are able to grow at
2-15 percent of concentrations)
Chemical Requirements: Along with water microbes also requires some other chemicals including,
1) Carbon (required to fulfill the needs of organic compounds which forms the living cell and is
important for the structure of cell)
2) Nitrogen and sulfur (important elements required for the synthesis of proteins and vitamins)
3) Phosphorous and nitrogen (required for making up the DNA and RNA and for synthesizing
the storehouse of energy called as ATP)
4) Potassium, Calcium and Magnesium (important elements because enzymes uses them as
cofactors)
5) Trace elements including zinc, molybdenum, copper and iron (generally found in tap water
and functions as cofactors)
6) Organic growth factors ( These factors must be provided to some bacteria’s which are
unable to synthesize on their own such as vitamins and other may be amino acids,
pyrimidines and purines)
7) Oxygen ( High oxygen concentration must be provided to obligate aerobes, Facultative
aerobes are able to grow anaerobically but best grows where oxygen resides, Obligate
anaerobes requires no oxygen and will stop growing if oxygen is provided, Aero tolerant
Anaerobes grows best without oxygen but can tolerate oxygen if present, Microaerophilic
grows when the concentration of oxygen is very low)
8) Superoxide Dismutase (used to prevent the toxicity caused by superoxide)

Q. No. 3. Give brief answers of the following questions; (Marks 03)

a. What is the logic behind presence of outer membrane in the cell wall of Gram negative bacteria
only?
 The outer membrane of gram negative bacteria contains Lipopolysaccharide (LPS) , a
complex molecule made up of three parts that are:
1) Lipid A (lipid part of lipopolysaccharide)
2) Core polysaccharide (composed of unusual sugar molecules)
3) O polysaccharide (composed of sugar molecules)
The Lipid A portion of lipopolysaccharide is released when the bacteria dies after causing
infection. Lipid A acts as an endotoxin which is involved in producing symptoms including
fever, clotting of blood, shock and blood vessels dilation Thus, symptoms caused by Lipid A
makes body aware of the infection. Also, this portion makes the bacteria virulent and less
susceptible to the antibiotics. Due to their outer membrane they are not penetrable.

b. Describe three reserve materials synthesized by microorganisms.

The cytoplasm of prokaryotic cells consists of storage bodies which are termed as inclusions.
These inclusions gather the nutrients and store them so that they may be used by cells in
time of need or when the environment becomes unfavorable. The three reserve materials
synthesized by microorganisms are:
1) Inorganic phosphate: Polyphosphate is another name used for this reserve. It is
present in metachromatic granules inclusions. This reserve is used in ATP synthesis
which acts as an energy source for cells. These inclusions are collectively called as
volutin. Methylene blue which is a blue dye stains these inclusions red.
2) Sulfur: This reserve is contained within the inclusions which are called sulfur
granules. Sulphur is synthesized by bacteria, which obtains energy by the oxidation
of sulfur and sulfur-containing compounds.
3) Iron Oxide: This reserve is present within the inclusions called Magnetosomes,
which are formed by magnet-like gram-negative bacteria such as Magnetospirillum
magnetotacticum.

c. Carl Woese suggested a phylogenetic classification scheme in 1978 that contained how many
domains? Describe these domains and explain the bases of his classification.
Carl woese in 1978 solved the classification problem of newly discovered organisms by
introducing a classification system for all organisms except for the viruses. He classified the
organisms into three domains on the basis of their cellular organization that is the
arrangement of different cellular components (presence or absence of nuclei, organelles
etc.). The three domains are Bacteria, Archaea and Eukaryotes.
1) Bacteria: All prokaryotic organisms having cell walls made up of peptidoglycan
(which is made up of proteins and carbohydrates) were included in this domain.
 2) Archaea: All prokaryotic organisms which lack peptidoglycan in their cell walls were
included in this domain. Polysaccharide (pseudomurein) is the component which
makes up their cell wall.
3) Eukaryotes: This domain consisted of Protists (All types of protozoa and algae were
included in this group), Fungi (It included all unicellular yeasts, multicellular yeasts
and Mushrooms), Plants ( It included trees, ferns etc.) and Animals( All vertebrates
including insects, flies etc. were included in this group).

d. Which molecules are responsible for the negative charge on bacterial cell?
Bacterial cell bears a negative charge. The molecules which imparts this negative charge differs in
gram-positive as well as in gram-negative bacteria.
1) Gram-positive bacteria: The negative charge on gram-positive bacteria is due to the teichoic
acids which are found either attached to the peptidoglycan or along the whole length of
peptidoglycan layer as lipoteichoic acid. Teichoic acid is made up of alcohol and phosphate.
Thus, the phosphate imparts the negative charge to the teichoic acid which in turn gives
negative charge to the bacterial cell.
2) Gram-negative bacteria: The negative charge on gram-negative bacteria is due to the
presence of lipopolysaccharides in their structure. These lipopolysaccharides are negatively
charged which imparts negative charge to bacterial cell.

Q. No. 4. Briefly state the role microorganisms play in each of the following; (Mark 01)
a. Biological control of pests
Microorganisms are effective in controlling the pests and bugs which attack crops, vegetation etc.
They completely kill the pests without affecting the nearby plants. Due to their specificity they are
used as an alternative for pesticides, insecticides. Their harmful effect on the environment is
minimal as compared with other pesticides. Their minute quantities can completely control pests.
Microorganisms which are involved in biological control of pests include protozoans, fungi, bacteria
and viruses.

b. Recycling of elements
Bacteria are the decomposers which are involved in the breakdown of dead organic material by
releasing enzymes thereby freeing up some elements such as carbons, potassium etc. Bacteria use
the organic carbon as their source of energy and the remaining elements are useful for plants.
Nitrogen fixing bacteria (cyanobacteria) convert the free nitrogen in the atmosphere into fixed
nitrogen, making it available for the plants. Bacteria are also involved in the nitrification in nitrogen
cycle such as Nitrosomonas and Nitrobacter. Then ammonifying and denitrification bacteria are also
involved in the nitrogen fixation cycle.

c. Human insulin production

Bacterial plasmid (which is an extrachromosomal DNA carrying antibiotic resistance genes) is widely
used as a vector in the laboratory to produce human insulin. The gene of interest is attached with
the plasmid (recombinant DNA) which then carries the gene of interest into the bacteria. Secondly,
bacterial cells are used as an expression system for the multiplication of the gene of interest.
Bacteria is chosen as expression system because its genetic makeup is simple and its generation time
is also small. Thus, bacteria are widely used in genetic engineering techniques.