Common Manual QUESSTIONS ALONE

AGM 101 Fundamentals of Microbiology
Questions in Practical record

1. Safety in microbiology laboratory - Study of Microscopes - Handling Light
Microscope
1. Write the purpose for which you will adjust the following parts of microscope
a. Diaphragm; b. Condenser; c. Fine adjustment; d. Stage control
2. Write the principle of using immersion oil in microscope
3. How will you differentiate SEM and TEM?
4. Define refractive index
5. Oil immersion objective lens is superior to dry objective lens. Why?
6. In a compound microscope, the objective lens has NA of 0.6 and the source lamp
has the light of 550 nm wave length. Calculate the resolving power (RP).
7. What is the maximum magnifying power of light microscope?
8. Why quartz lenses are used in place of glass lenses in UV microscope?
2. Micrometry‐Measurement of Microorganisms
1. How can you differentiate the stage micrometer and ocular micrometer?
2. We have to do calibration for every objective lens. Why?
3. If one stage micrometer division equals 17 ocular divisions, calculate the
calibration factor.
4. If one stage micrometer division occupies 22 ocular divisions and a bacterial cell
occupies 3 ocular divisions, calculate the length of the bacterium.
5. You got different lengths under different objectives (10x and 40x) viewing same
cell. Explain the possible reason.
3. Aseptic techniques – working with different equipment

1. What do you mean by sterilization and sterilant?
2. What is the temperature required for sterilizing in hot air oven?
3. What is the standard sterilization temperature in autoclave?
4. Why moist heat is better than dry heat?
5. What is the mechanism of action of moist heat?
6. What are all the sterilizing agents used in Laminar Air Flow Chamber?
7. Why heat labile substances can’t be sterilized in hot air oven?
8. What is the mechanism of action of chemicals viz., ethanol, mercuric chloride,
ethylene oxide and formaldehyde?
4. Preparation of Culture media

1. Why do we need to prepare medium for microorganisms?
2. Name the carbon and nitrogen sources of Nutrient agar medium.
3. What is the significance of adding streptomycin to RBA?
4. What are the advantages of defined, synthetic and complex medium?
5. What is the principle behind the use of cotton plugs?
6. What is the significance of leaving air space in the flasks while preparing media?
7. Why agar is preferred as solidifying agent than gelatin?
8. Whether the pH of the medium will be changed after sterilization? Why?
9. Can we sterilize the media in hot air oven? Substantiate.
5. Isolation of microorganisms by Serial dilution plate technique

1. Why higher dilution sample is used for bacterial enumeration?
2. What is the role of Rose Bengal stain?
3. Why we use Streptomycin during fungal enumeration?
4. What will you do if no bacteria found in 10-6 dilution?
Purification of Bacteria and Actinobacteria

1. Name the techniques used for purifying bacteria.
2. How frequent a culture is sub cultured when storing at refrigerated conditions?
3. What is the role of cryopreservant?
4. What is the temperature of solid carbon and liquid nitrogen?
5. Name the equipment used for freezing and thawing of culture.
6. Illustrate the way to retrieve cultures from cryoprotected vials.
6. PURIFICATION OF YEASTS AND MOLDS

1. List the ways to purify molds and yeasts.
2. On what basis you will decide the appropriate purification method of molds?
7. PRESERVATION OF BACTERIA, FUNGI AND ACTINOBACTERIA

----------
8. STAINING TECHNIQUES‐ POSITIVE AND NEGATIVE STAINING
1. What is the need to stain bacteria?
2. Basic dyes are commonly used to stain bacteria. Why?
3. Is heat fixation necessary for proper staining? Reason out?
4. Why the background rather than the cells got stained in negative staining?
5. Give examples for basic, acidic and neutral dyes.
9. DIFFERENTIAL STAINING ‐ GRAM STAINING

1. Describe the function of each of the following in the Gram stain
a. Mordant b. Primary stain c. Decolorizer d. Counterstain:
2. Which step in the Gram stain is most likely to cause poor results if done
incorrectly?
3. Why must fresh bacterial cultures be used in a Gram stain?
4. Briefly describe the mechanism of Gram staining.
10 Turbidometric assessment of growth of bacteria

.
1. Is it possible to get decreased OD values against time course? Why?
2. Why variations occur in generation time among the bacterial species?
3. The growth curve of bacteria will be same at different carbon source used.
True or false. Explain?
4. What are the factors that influence the growth curve of bacteria?
5. Is it possible to extend the log phase of bacteria by any means?
6. In which phase bacterial growth will be more?
7. Is it possible to get decreased OD values against time course? Why?
11 Morphological characteristics of bacteria and actinobacteria

.
1. Classify bacteria based on shape and arrangement with examples
2. List out the colony and cell morphological characters used as criteria for
identification
12 Biochemical Characteristics of Bacteria and Actinobacteria
.
1. Distinguish between aerobic, anaerobic and fermentative modes of respiration
2. Explain the mechanism behind sherry red compound formation in Indole test
3. Why the color of the medium changes from green to blue in citrate utilization
test?
4. Illustrate the mechanism involved in the degradation of H 2O2 in the presence of
catalase
5. How can gas production be detected in carbohydrate fermentation test?
13 Identification of Yeasts, Molds and Algae - Morphological characterization

.
1. Name two pigment producing fungi.
2. Name the fungi producing sickle shaped conidia.
3. List some of the culture collection centres for fungi and algae.
4. Describe the features of heterocyst.
5. What are different types of sexual and asexual spores produced by fungi?
6. Give examples for heterocystous and non heterocystous blue green algae.
7. Differentiate green algae from blue green algae.
8. List out the products of industrial and agricultural importance from fungi and
algae.
14 Isolation of Bacteriophages
.
1. What type of phages can be isolated from this experiment, lytic or lysogenic
phages?
2. Can temperate phages be demonstrated in this experiment? Why.
3. What is the principle behind plaque formation in the bacterial plates?
4. Why do you need to inoculate the phage samples collected from any environment
into bacteria before isolating them?
15 Development of bacterial mutants by employing UV Irradiation / Chemical

. mutagens.
1. Define mutation
2. How mutations occur in bacteria?
3. What is a mutagen? Give several examples.

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AGM 101 Fundamentals of Microbiology

Questions in Practical record

3. Aseptic techniques – working with different equipment

4. Preparation of Culture media

5. Isolation of microorganisms by Serial dilution plate technique

Purification of Bacteria and Actinobacteria

6. PURIFICATION OF YEASTS AND MOLDS

7. PRESERVATION OF BACTERIA, FUNGI AND ACTINOBACTERIA

9. DIFFERENTIAL STAINING ‐ GRAM STAINING

10 Turbidometric assessment of growth of bacteria

11 Morphological characteristics of bacteria and actinobacteria

13 Identification of Yeasts, Molds and Algae - Morphological characterization

15 Development of bacterial mutants by employing UV Irradiation / Chemical

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