Exercise 5: Case Studies Flashcards

1
Q

If you receive a swab from a wound, pus, bite, or from the eye or ear, and the bacteria in question is known to survive in aerobic conditions, which medias would you perform the preliminary cultures on?

A
  1. HBA
  2. CBA
  3. MAC
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2
Q

Give the name of a selective media that you would use to isolate faecal samples

A

MAC (MacConkey agar)

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3
Q

Describe MacConkey agar

A
  • Is considered both selective and differential
  • Selectively isolates gram negative and enteric bacilli (the bile salts are selective for enteric organisms)
  • Differentiates these enteric organisms based on lactose fermentation ->
    Lactose fermenter = PINK
    Non-lactose fermenter = CLEAR
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4
Q

Describe the features of S. aureus

A

Morphology:

  • Gram positive
  • Cocci
  • Clusters

Media & details on media:

  • HBA - forms yellow colonies, beta haemolytic
  • CBA - forms yellow colonies
  • MAC - no colonies formed (as the bile salts are selective for gram negative enteric organisms)
  • MSA - forms yellow colonies

Test results:

  • Catalase +
  • Bactistaph +
  • All Staphylococci are facultative anaerobes

Key virulence factors:

  • Produces toxins (haemolysins, leukotoxins, leukocidin)
  • Catalase (enhances survival in phagocytes)
  • Invasins (hyaluronidase)

Effects of infection:

  • Toxic shock syndrome
  • Septic shock

Treatment:
- Antibiotics - Vancomycin and Tetracycline

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5
Q

Describe the features of Streptococcus pneumoniae.

A

Morphology:

  • Gram positive
  • Cocci
  • Paired

Media & details on media:

  • HBA - forms colonies, alpha haemolytic
  • CBA - forms colonies
  • MAC - no colonies formed (as the bile salts are selective for gram negative enteric organisms)
  • Facultative anaerobe

Key virulence factors:

  • CAPSULE
  • Invasins
  • Adhesins
  • Exotoxins

Effects of infection:

  • Pneumonia
  • Meningitis
  • Bronchitis

People most susceptible to infection:

  • Elderly people
  • Children under the age of 5
  • Immunocompromised individuals

Treatment:

  • Pneumococcal vaccines
  • Antibiotics - Penicillin
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6
Q

How do you confirm a S. pneumoniae infection?

A
  1. Obtain a swab from a patient and perform a gram stain (should observe Gram positive cocci in a paired arrangement)
  2. Culture on a HBA plate (should observe alpha haemolysis)
  3. Place a optochin disc on the HBA (should be sensitive to optichon)
  4. Capsule stain (Maneval’s stain) to confirm that it is S. pneumoniae
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7
Q

Define what a fastidious organism is

A
  • An organism that has complex nutritional requirements
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8
Q

Describe what CBA is

A
  • Chocolate blood agar
  • Is non-selective
  • A variant of the blood agar plate, containing red blood cells which have been lysed slowly
  • Used for growing fastidious respiratory bacteria, such as Haemophilus influenzae
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9
Q

Describe the oxidase test

A
  • Used to identify bacteria that produce cytochrome c oxidase (enzyme of the bacterial electron transport chain)
  • Positive results indicate that the bacteria is aerobic

+ve test = dark purple colour
-ve test = no colour change

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10
Q

How do you differentiate between Staphylococcus and Streptococcus?

A
  • Catalase test

- Staphylococci are catalase positive, while Streptococci are catalase negative

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11
Q

Describe the features of Streptococcus pyogenes

A

Morphology:

  • Gram positive
  • Cocci
  • Short chains

Media & details on media:

  • HBA - forms colonies, beta haemolytic
  • CBA - forms colonies
  • MAC - no colonies formed (as the bile salts are selective for gram negative enteric organisms)

Test results:

  • Catalase -
  • Facultative anaerobe

Key virulence factors:

  • M protein
  • Hyaluronidase
  • Exotoxins (Streptolycin O, Streptolycin S)
  • Invasins (Streptokinase)

Effects of infection:

  • Toxic shock syndrome
  • Scarlet fever

Treatment:
- Antibiotics - Penicillin

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12
Q

How do you confirm a S. pyogenes infection?

A
  1. Gram stain (should observe gram positive cocci in short chains)
  2. Perform a catalase test (should be catalase negative)
  3. Observe colonies on HBA plate (should be beta haemolytic)
  4. Place bacitracin disc on HBA (should be bacitracin sensitive)
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13
Q

Describe the features of Salmonella enterica

A

Morphology:

  • Gram negative
  • Rods
  • Flagellated (motile)

Media & details on media:

  • MAC - clear colonies formed (ie lac negative)
  • XLD - red colonies with black centres

Test results:

  • Oxidase -
  • Facultative anaerobe

Key virulence factors:

  • Type III secretion system
  • Flagella
  • Invasins
  • Enterotoxins

Effects of infection:

  • Typhoid
  • Gastroenteritis

Treatment:
- Antibiotics

Prevention:

  • Hand washing
  • Cooking food well
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14
Q

How do you confirm an infection by Salmonella enterica?

A
  1. Grow on MAC agar (should be Lac negative - Clear)
  2. Perform an oxidase test (should be oxidase negative)
  3. Grow on XLD agar (should be red colonies with BLACK centres)
  4. Identify using agglutination (should agglutinate)
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15
Q

Describe Vibrio cholerae

A

Morphology:

  • Gram negative
  • Rods
  • Flagellated (motile)

Media & details on media:

  • MAC - clear colonies formed (ie lac negative)
  • TCBS - yellow colonies

Test results:

  • Oxidase +
  • Facultative anaerobe

Key virulence factors:

  • Cholera toxin
  • Flagella
  • Pilus

Effects of infection:
- Cholera

Treatment:
- Antibiotics

Prevention:

  • Water purification
  • Proper sanitation
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16
Q

How do you confirm an infection by Vibrio cholera?

A
  1. Grow on MAC agar (should be Lac negative - Clear)
  2. Perform an oxidase test (should be oxidase positive)
  3. Grow on TCBS agar (should observe yellow colonies)
17
Q

Describe XLD agar

A
  • Xylose lysine deoxycholate agar
  • Used to isolate Salmonella and Shigella species

Red colonies with black centres - Salmonella (forms black spot due to the production of H2S)

Red colonies - Shigella

Yellow colonies - normal gut flora

18
Q

Describe TCBS agar

A
  • Thiosulfate citrate bile sucrose agar
  • Selective media for pathogenic Vibrio sp.

Yellow - Vibrio cholera
Green - Other Vibrios

19
Q

Describe Mycobacterium tuberculosis

A

Morphology:

  • Can appear gram poitive or negative
  • Small rod

Media & details on media:
- Lowenstein jensen media - brown colonies

Tests:

  • Acid fast +
  • Catalase +
  • Strict aerobe

Key virulence factors:

  • Mycolic acid
  • Slow generation time
  • Intracellular growth

Effects of infection:
- Tuberculosis

Treatment:
- Drugs

20
Q

How do you confirm an infection by M. tuberculosis?

A
  1. Perform an acid-fast stain (should be red)
  2. Observe on Lowenstein-jensen medium (3-4 weeks incubation due to slow doubling time of TB; should observe brown colonies)
21
Q

What are acid fast stains?

A
  • Ziehl Neelsen or Kinyoun’s stain
    Differential stains used to detect acid- and alcohol- fast organisms
  • Useful for detection of Mycobacterium sp.

Red - acid fast organisms
Blue/green - non acid fast organisms