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Methods in Microbiology

Description: Microbiology GATE Stain Life Sciences Microorganisms Gram staining Acid fast staining Agar Voges-Proskauer test Indole test Urease test Catalase test Methyl red test Microbiology
Number of Questions: 15
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Tags: Microbiology GATE Stain Life Sciences Microorganisms Gram staining Acid fast staining Agar Voges-Proskauer test Indole test Urease test Catalase test Methyl red test
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Which of the following are used as counterstain in Gram staining technique? P. Crystal violet Q. Safranin R. Iodine solution S. Carbol fuchsin Select the correct answer.

  1. P and Q

  2. P and R

  3. Q and R

  4. Q and S

  5. R and S


Correct Option: D
Explanation:

Safranin is used as secondary stain (counterstain) in Gram staining technique. Carbol fuchsin is sometimes substituted for safranin as counterstain, since it  more intensely stains anaerobic bacteria.

Gram staining is a common technique used to differentiate Gram-positive and Gram-negative bacteria based on their different cell wall constituents. Colours of Gram-positive and Gram-negative bacteria on Gram staining respectively are _________.

  1. pink, purple

  2. purple, pink

  3. purple, colourless

  4. colourless, purple

  5. colourless, pink


Correct Option: B
Explanation:

The Gram stain technique distinguishes between Gram-positive and Gram-negative bacteria by colouring these cells purple or pink. Gram positive bacteria stains purple due to the presence of a thick layer of peptidoglycan in their cell walls, which retains the crystal violet these cells during the decolouring process. Gram negative bacteria stains pink, which is attributed to a thinner peptidoglycan wall, which does not retain the crystal violet during the decolouring process.

Which of the following is/are example(s) of both selective media as well as differential media?

  1. Nutrient Agar
  2. Mannitol Salt Agar
  3. MacConkey agar
  1. Only 1

  2. Only 2

  3. Only 3

  4. Only 1 and 2

  5. Only 2 and 3


Correct Option: E
Explanation:

Mannitol salt agar is both a selective and differential media used for the isolation of pathogenic Staphylococci from mixed cultures. MacConkey’s Agar is both a selective and differential media. It is selective for Gram-negative bacteria and can differentiate those bacteria that have the ability to ferment lactose.

Which of the following are bacteriological stains used to identify acid-fast organisms?

  1. Auramine-rhodamine stain
  2. Gram's stain
  3. Hoechst stain
  4. Kinyoun stain
  5. Ziehl-Neelsen stain Select the correct answer.
  1. 1, 2 and 3

  2. 1, 3 and 5

  3. 1, 4 and 5

  4. 2, 3 and 4

  5. 3, 4 and 5


Correct Option: C
Explanation:

Auramine-rhodamine stain is a histological technique used to visualize acid-fast bacilli using fluorescence microscopy. Kinyoun stain is a method of staining acid-fast microbes, specifically Mycobacterium and Nocardia. Ziehl-Neelsen stain is used to identify the acid-fast bacteria mainly Mycobacterium.

Which of the following staining methods uses carbol fuchsin and methylene blue as primary stain and counterstain respectively?

  1. Gram's Stain
  2. Kinyoun stain
  3. Ziehl–Neelsen stain
  1. Only 1

  2. Only 2

  3. Only 3

  4. 1 and 2

  5. 2 and 3


Correct Option: E
Explanation:

The reagents used in Ziehl–Neelsen staining method are carbol fuchsin (primary stain), acid alcohol, and methylene blue (counterstain). The Kinyoun staining method uses carbol fuchsin as a primary stain, followed by decolourization with an acid-alcohol solution and methylene blue as a counterstain.

Which of the following test determines microbial ability to perform mixed-acid fermentation?

  1. Citrate test

  2. Voges-Proskauer test

  3. Methyl red test

  4. Indole test

  5. Rapid urease test


Correct Option: C
Explanation:

Methyl red test detects production of acids formed during metabolism using mixed acid fermentation. The pH indicator Methyl Red is added to one tube and a red colour appears at pH's lower than 4.2, indicating a positive test (mixed acid fermentation is used). The solution remaining yellow (pH = 6.2 or above) indicates a negative test, meaning the butanediol fermentation is used.

Examples of enriched media are ____________.

  1. hektoen enteric agar, mannitol salt agar

  2. eosin Methylene Blue agar, macConkey agar

  3. blood agar, chocolate agar

  4. eosin Methylene Blue agar, mannitol salt agar

  5. blood agar, hektoen enteric agar


Correct Option: C
Explanation:

Blood agar is both differential and enriched medium. The blood that is incorporated into this medium is an enrichment ingredient for the cultivation of fastidious organisms such as the Streptococcus species. Chocolate agar is enriched culture media commonly used for primary isolation of Haemophilus species from clinical specimens.

Which of the following tests can be used to differentiate between Proteus mirabilis and Proteus vulgaris?

  1. Urease test

  2. Phenylalanine deaminase test

  3. Indole test

  4. Methyl red test

  5. Catalase test


Correct Option: C
Explanation:

Proteus mirabilis and Proteus vulgaris can be differentiated by an indole test. Proteus vulgaris tests positive for indole test while Proteus mirabilis test negative for indole test.

Which of the following are examples of Gram-negative bacteria?

  1. Alcaligenes faecalis
  2. Bacillus subtilis
  3. Enterobacter aerogenes
  4. Staphylococcus aureus
  5. Klebsiella pneumoniae
  6. Streptococcus pyogenes
  1. 1, 2 and 3

  2. 1, 3 and 5

  3. 2, 3 and 5

  4. 2, 4 and 6

  5. 3, 4 and 5


Correct Option: B
Explanation:

Alcaligenes faecalis, Enterobacter aerogenes, Klebsiella pneumoniae: All are Gram-negative bacteria.

Which of the following sets represents catalase-positive bacteria only?

  1. Enterobacter, Klebsiella, Nocardia, Staphylococcus

  2. Enterococcus, Klebsiella, Nocardia, Streptococcus

  3. Enterobacter, Enterococcus, Staphylococcus, Streptococcus

  4. Enterococcus, Klebsiella, Nocardia, Staphylococcus

  5. Klebsiella, Nocardia, Staphylococcus, Streptococcus


Correct Option: A
Explanation:

Catalase Test: The catalase test is done by placing a drop of hydrogen peroxide on a microscope slide. If the bacteria possess catalase, when a small amount of bacterial isolate is added to hydrogen peroxide, bubbles of oxygen are observed. If the mixture produces bubbles or froth, the organism is said to be 'catalase-positive' otherwise it is 'catalase-negative’. Enterobacter, Klebsiella, Nocardia, and Staphylococcus: All are catalase-positive bacteria.

Which of the following tests can be used in the differentiation of Staphylococcus aureus and Streptococcus pyogenes?

  1. Indole test

  2. Voges-Proskauer test

  3. Cytochrome C oxidase test

  4. Catalase test

  5. All of the above


Correct Option: D
Explanation:

Catalase test is used to detect the presence of the enzyme catalase in bacteria. Catalase serves to neutralize the bactericidal effects of hydrogen peroxide. It is a primary test used in the differentiation of Staphylococci and Streptococci. Staphylococcus aureus tests positive while Streptococcus pyogenes tests negative for catalase test.

All of the following are the examples of differential media except
P. Blood agar Q. Chocolate agar R. Eosin methylene blue agar S. Hektoen Enteric Agar T. Phenylethyl alcohol agar

  1. P and Q

  2. Q and R

  3. Q and T

  4. R and S

  5. S and T


Correct Option: C
Explanation:

Chocolate agar: Enriched media Phenylethyl alcohol agar: Selective media.

Which of the following mediums are selective for Gram-negative bacteria? P. Hektoen enteric agar Q. MacConkey agar R. Mannitol salt agar S. Yeast Malt Agar

  1. P and Q

  2. P and S

  3. Q and R

  4. Q and S

  5. R and S


Correct Option: A
Explanation:

Hektoen Enteric Agar is a selective and differential agar for the isolation and cultivation of gram-negative enteric microorganisms, especially Salmonella and Shigella. MacConkey Agar is a selective and differential medium designed to isolate and differentiate Gram-negative enterics based on their ability to ferment lactose.

Which of the following statements are FALSE regarding Transmission Electron Microscopy (TEM)?

  1. First Transmission Electron microscope was built by Knoll and Ruska.
  2. The electronic beam is focused by electromagnetic lenses.
  3. TEM is capable of imaging at a significantly higher resolution than light microscopes.
  1. Only 1

  2. Only 2

  3. Only 3

  4. 2 and 3

  5. None


Correct Option: E
Explanation:

All are true statements. First TEM was built by Max Knoll and Ernst Ruska in 1931. TEMs are capable of imaging at a significantly higher resolution than light microscopes, owing to the small de Broglie wavelength of electrons. In TEM electrons are accelerated by an electric potential and focused by electrostatic and electromagnetic lenses onto the sample.

Which of the following pairs of bacteria shows positive Voges-Proskauer test?

  1. Enterobacter aerogenes and Escherichia coli

  2. Enterobacter aerogenes and Proteus mirabilis

  3. Escherichia coli and Klebsiella pneumoniae

  4. Escherichia coli and Proteus mirabilis

  5. Enterobacter aerogenes and Klebsiella pneumoniae


Correct Option: E
Explanation:

Voges–Proskauer test is used to detect acetoin in bacterial broth culture. The test is performed by adding alpha-naphthol and potassium hydroxide to the Voges-Proskauer broth,  which has been inoculated with bacteria. A cherry red colour indicates a positive result, while a yellow-brown color indicates a negative result. Both Enterobacter aerogenes and Klebsiella pneumoniae give positive Voges–Proskauer test.

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