Maizatul Mohd.

MLT/DP/10-3/0027

MICROBIOLOGY

Short Question

1. What are the instrument used to sterilize serum and air containing medium?
Hot air oven

2. What is a smear?
A sample spread over a microscope slide to be studied for any pathology
eg. blood sample
types
thin - to study the morphology, blood is spread over the glass slide, examination
of parasite, type of parasite
thick - blood is drop and cover with cover slip (no spreading), examination of
malaria, detection of malaria
wet - to study the motility

3. What is the function of the illuminator?

Light source, electric light provided by a lamp built inside the microscope beneath
the stage

4. How should you fix the smear on the glass slide?

By using heat fixation method, after a smear has dried at room temperature, the
slide is gripped by tongs or a clothespin and passed through the flame of a Bunsen
burner several times to heat-kill and adhere the organism to the slide. Routinely
used with bacteria and archaea. Heat fixation generally preserves overall
morphology but not internal structures. Heat denatures the proteolytic enzyme and
prevent autolysis. Heat fixing helps attach the cells to the slide so they are not
washed off during the staining process, kills the cells so the slide is not hazardous
to handle, and alters the cell wall for staining.

5. When would you use the wet mount technique?

To study the motility of microscopic organism, studying their movement and
behavior. It is also used to examine physiological liquids like blood, urine, saliva,
semen, and vaginal discharge. Care must be taken to exclude air bubbles that
would interfere with the viewing and hamper the organisms' movements. Wet
mount microscope slides are used with live organisms when the observer needs to
view the subject while it is in motion or is reacting to some sort of environmental
stimulus.

Maizatul Mohd. MLT/DP/10-3/0027

Maizatul Mohd. MLT/DP/10-3/0027

6. What is an inoculating loop/needle?

Inoculating loops are used to transfer microorganisms to growth media or for
staining slides. They are an important part of the sterile technique as their use
permits transfer only of the material of interest. The loop is also used in the
cultivation of microbes on plates by transferring inoculum for streaking. The
inoculation loop is sterilized in a flame until it becomes red hot before and after
each use. By doing this, the same tool can be reused in different experiments
without fear of cross-contamination. After flame sterilization, the loop must be
cooled so that the next cells the loop touches are not killed by the hot metal.

7. Microscope Parts & Function

a) supporting
- body tube: tube with objective lens at the lower end, eyepiece at the upper end
- revolving nose piece: rotating disc, having holes for objective lens, can be
rotated to change the power of magnification
- stage: placement of slide, have hole in the center to allow light to fall on the
specimen through the hole
- arm: holds body tube and coarse adjustment
- coarse adjustment: moves stage up and down (adjust the focal length)
- fine adjustment: to obtain exact focussing

b) illuminating
- illuminator: light source, electric light provided by lamp built inside the
microscope beneath the stage
- reflecting mirror: reflects light
- condenser: focuses the reflected light
- iris/diaphragm: allows the required amount of light to pass through

c) magnification
- eyepiece/ocular lens: allows observer to look the magnified image through the
body tube
- objective lens: magnifies the specimen

d) other parts
- stage clips: holds the slide
- coxial stage control: to adjust the stage left/right and up/down (horizontally)
- brightness control: to adjust the brightness of the light
- switch: on/off the microscope

Maizatul Mohd. MLT/DP/10-3/0027

Maizatul Mohd. MLT/DP/10-3/0027

8. What part of compound microscope regulates the amount of light passing through
the slide specimen on the microscope stage?
Iris/diaphragm

9. Why must small object seen through under ocular lens be centered in the field
view before changing to higher objective lens?
- when the magnification increases, the lens will zoom in and cut the periphery
view of the specimen
- as the magnification increases, the diameter of the field of view decreases. this
phenomenon will become more important when we use compound microscopes,
and is often the reason that objects or structures seemingly “disappear” when the
magnification is increased
- in other words, if an object is not centered in the field of view under low
magnification, it may not appear in the (more limited) field of view under higher
magnification

10. Why do you adjust if you can’t see two overlapping circle with parts of the object
in each circle?
Change the distance between the ocular lens

11. Who is the father of microbiology?

Louis Pasteur

Maizatul Mohd. MLT/DP/10-3/0027

Maizatul Mohd. MLT/DP/10-3/0027

12. Who is the father of bacteriology?

Robert Koch

13. Who discovered the antiseptic method?

Joseph Lipster (father of surgery)

14. Which substance is used to sterilize the work bench?

Phenol

15. How do you find out whether the bacteria is gram-positive or gram- negative?
- Gram-positive bacteria appear blue or violet
- Gram-negative bacteria appear pinkish red

16. Which is the ideal place to do bacterial culture?

LAMINAR air flow

17. Define the following terms:

a) bactericidal - refers to a substance (or a condition) capable of killing bacteria.
Preventing infection by inhibiting the growth or action of microorganisms

b) antiseptic - an antiseptic is a substance which inhibits the growth and

development of microorganisms. An agent that kills or prevents the growth of
bacteria.

c) disinfectant - an agent, such as heat, radiation, or a chemical, that disinfects by

destroying, neutralizing, or inhibiting the growth of disease-carrying
microorganisms. An agent that destroys infection-producing organisms. Heat and
certain other physical agents such as live steam can be disinfectants, but in
common usage the term is reserved for chemical substances such as mercury
bichloride or phenol. Disinfectants are usually applied to inanimate objects since
they are too strong to be used on living tissues. Chemical disinfectants are not
always effective against spore-forming bacteria.

d) disinfection - the process of destroying pathogenic organisms or rendering

them inert. The process or act of destroying pathogenic microorganisms.

e) bacteriostatic - preventing bacteria from growing and multiplying but possibly

not killing them. Arresting the growth or multiplication of bacteria; also, an agent
that so acts. A term describing substances such as sulfonamides and tetracycline
which inhibit the growth and propagation of bacteria, but do not actually destroy
bacteria.

18. In which color appear the gram-positive bacteria at the end of staining procedure?
Blue

Maizatul Mohd. MLT/DP/10-3/0027

Maizatul Mohd. MLT/DP/10-3/0027

19. Arrangement of bacteria:

Cocci are spherical or oval shaped bacteria
a) streptococci – linear arrangement of cocci bacteria
b) staphylococci – in a cluster arrangement of cocci bacteria
c) corny bacterium – rod-like shaped (bacilli)
d) vibria cholera – spiral-like shaped (spirillum, vibrio, spirals)
e) neilsen gonorrhoeae – irregular shaped

20. What is the holding time and temperature for hot air oven?
Temperature (°C) Time (minutes)
120 480
140 180
150 150
160 120
170 60
180 30

21. What is the holding time and temperature of autoclave?

120-121°C for 15-20 minutes

22. What kind of filter present in the LAMINAR air flow chamber?
HEPA filter

23. What is HEPA filter?

High efficiency particulate air

24. What is the function of HEPA filter?

HEPA filter acts like a sieve where particles smaller than the largest opening can
pass through is incorrect. Unlike membrane filters, where particles as wide as the
largest opening or distance between fibers cannot pass in between them at all,
HEPA filters are designed to target much smaller pollutants and particles. HEPA
filters are critical in the prevention of the spread of airborne bacterial and viral
organisms and, therefore, infection. Typically, medical-use HEPA filtration
systems also incorporate high-energy ultra-violet light units to kill off the live
bacteria and viruses trapped by the filter media. Some of the best-rated HEPA
units have an efficiency rating of 99.995%, which assures a very high level of
protection against airborne disease transmission.

25. Who conducted the experiment with swan-neck flask?

Louis Pasteur

26. Which bacteria is not having peptidoglycan in the cell wall?

Gram-negative bacteria

Maizatul Mohd. MLT/DP/10-3/0027

Maizatul Mohd. MLT/DP/10-3/0027

27. What is a mordant?

A chemical compound which reacts with the stain to form an insoluble, coloured
precipitate. When excess dye solution is washed away, the mordanted stain
remains.

28. Give one example of a mordant?

Iodine crystals

29. What are the uses of mordant?

Mordant is used in gram staining. Mordant is used as fixation to intensifying or
deepening the staining action of crystal violet to the organism.

30. What is the primary stain used in the gram staining procedure?
Crystal violet

31. What is the counter stain used in the gram staining procedure?
Safranin

32. What is animalicules?

Microscopic organism such as an amoeba or paramecium. An animal, invisible, or
nearly so, to the naked eye.

33. Who used the word animalicules?

Anthony Van Leeuwenhoek

34. Who proposed the Koch’s Postulate?

Robert Koch

35. Describe Koch’s Postulates

a) Bacteria should always be present with the lesions of the disease
b) it should be possible to isolate the bacterium in pure culture from the lesion
c) when we inoculate the above culture into lab animals, it should reproduce the
lesions of the disease
d) it should be possible to reisolate the bacterium in pure culture from the lesions
produced in the experimental animals

36. What is the meaning of pasteurization?

Pasteurization is a process of heating a food, usually liquid, to a specific
temperature for a definite length of time, and then cooling it immediately. This
process slows microbial growth in food. Pasteurization is not intended to kill all
micro-organisms in the food. Instead pasteurization aims to reduce the number of
viable pathogens so they are unlikely to cause disease.

37. What is the temperature prepared for pasteurization?

63°C for 30 minutes

Maizatul Mohd. MLT/DP/10-3/0027