Microbiology Lab continues28 cards

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1

Pseudomonas Aeruginosa

STAIN:
    -Gram: negative

2

Pseudomonas Fluorescens

COMMENTS:
     -Monotrichous

3

Proteus Vulgaris

SPECIAL MEDIA:
      -MCA: Colorless
      -EMB: Green/Purple

4

Proteus Mirabilis

SPECIAL MEDIA:
      -PEA: no growth
COMMENTS:
      -Peritrichous, motile

5

Bacillus Stearothermophilus

STAIN:
    -Spore: sub-terminal
COMMENTS:
    -Aerobes

6

Bacillus Anthracis

STAINS:
     -Spore: Yes
DISEASE:
     -Anthrax

7

Bacillus Cereus

STAIN:
    -Spore: yes
DISEASE:
    -food poisoning

8

Clostridium Sporgenes

STAIN:
     -Spore: terminal
COMMENTS:
     -Anaerobes

9

Clostridium Tetani

STAIN:
     -Spore: yes
DISEASE:
     -Tetanus

10

Clostridium Perfringens

STAIN:
     -Spore: yes
DISEASE:
     -Gas gangrene
COMMENTS:
     -also food poisoning

11

Clostridium Botulinum

STAIN:
     -Spore: yes
DISEASE:
     -Botulism

12

Spirillum Volutans

COMMENTS:
     -Amphitrichous

13

Serratia Marcescens

DISEASE:
     -Nosocomial infections
COMMENTS:
     -at 25-30 degrees C, it produces a red pigment: "Prodifiosin"
     -White at higher temperatures

14

Escherichia Coli

SPECIAL MEDIA:
     -MSA: no growth
     -HEA: Salmon-pink
DISEASE:
      -Food poisoning
COMMENTS:
      - Cream colored

15

Micrococcus Luteus

COMMENTS:
     -Yellow due to carotenoid pigments

16

Enterococcus Faecalis

SPECIAL MEDIA:
     -PEA: growth
     -HEA: no growth

17

Salmonella Typhimurium

SPECIAL MEDIA:
       -HEA: black

18

Phenyl Ethyl Alcohol (PEA)

PURPOSE:
     -To isolate streptococci & staphylococci from mixed cultures
COMPOSITION:
     -contains phenyl ethy alcohol
FUNCTION:
     -Inhibits Gram Negative bacteria (Proteus Mirabillis)
     -Selects for Gram Positive bacteria
RESULT:
     -Growth: Gram positive (Staph. Epidermidis and Enterococcus Faecalis)
     -No growth: Gram Negative (Proteus Mirabillis)

19

Mannitol Salt Agar

PURPOSE:
      -to isolate and differentiate pathogenic organisms like staphylococcus aeurus
COMPOSITION:
      -Mannitol, 7.5% NaCl (organisms cant grow in this amount of salt), and Phenol Red
FUNCTION:
      -Inhibits most organisms
      -fermentation substrate
      -detects pH levels (3 levels)
      -Selects for staphylococcus
RESULTS:
      -No growth--->not a staphylococcus (E. coli)
      -Yellow--> strong fermenter (pH<6.8) (Staph. Aeurus)
      -Red--->slow fermenter (7.4<pH<8.4)
      -Pink--->Non-fermenter (pH>8.4) (staph. epidermidis).

20

MacConkey Agar (MCA)

PURPOSE:
      -to select & differentiate members of Enteriobacteria
COMPOSITION:
      -Lactose, Bile salts, Neutral Red, Crystal violet
FUNCTION:
      -inhibits Gram positive bacteria
      -fermentation substrate
      -detects pH levels (2 levels)
      -Selects for Gram Negative bacteria
RESULT:
      -No growth---> Gram Positive (staph. epidermidis)
      -Red---> Lactose Fermenter (pH<6.8) (Klebsiella Pneumonia)
      -Colorless---> Non-fermenter (pH>6.8) (Proteus Vulgaris)

21

Eosin Methylene Blue (EMB)

PURPOSE:
       -to isolate fecal coliforms
COMPOSITION:
       -eosin, methylene blue, lactose, sucrose, peptone
FUNCTION:
       -differentiates on the basis of fermetation
       -inhibits Gram Positive bacteria
       -selects for Gram Negative bacteria
       -fermentation substrate
       -nutrition source
RESULT:
       -No growth---> Gram Positive (staph. aureus)
       -Green metallic/deep purple-->fecal coliforms (Proteus Vulgaris)
       -Pink--> slow fermenter (low acid production) (Klebsiella Pneumonia)
       -colorless-->non-fermenters

22

Hektoen Enteric Agar (HEA)

PURPOSE:
      -to isolate salmonella and shigella species from other enterics
COMPOSITION:
      -Bile salts, peptone, lactose, sucrose, salicin, Na Thiolsulfate, Fe Ammonium Citrate, Bromothymol Blue & Acid Fuchsin
FUNCTION:
      -inhibits Gram Positive bacteria
      -selects for Gram Negative bacteria
      -Nutrition source
      -fermentation substrates that promote the growth of fecal coliforms
      -source of sulfur
      -reduces S in H2S to form a black precipitate
      -detects pH levels, differentiates on the basis of fermentation of sugars and production of acids
RESULT:
      -No growth--->Gram Positive (Enterococcus Faecalis)
      -Black--->Sulfur reducer (Salmonella Typhimurium)
      -Blue-Green--->Non-fermenter
      -Yellow to Salmon-Pink-->sugar fermenters, acid producers (E. Coli)

23

Negative Stain

PRIMARY STAIN:
      -Nigrosine or India Ink
RESULTS:
      -Acidic dye (- Chromophores) do not penetrate organism (- surface)
      -It stains the background only
PURPOSE & COMMENTS:
      -to observe cell morphology and size
      -no heat fixing-->no cell distortion or shrinkage

24

Capsular Stain

PRIMARY STAIN:
      -Congo Red
DECOLORIZER:
      -Water
SECONDARY STAIN:
      -Maneval's Stain
RESULTS:
      -Background stained with congo red
      -Cell proper stained with Maneval's Stain
      -Capsule is colorless
PURPOSE & COMMENTS:
      -to detect the presence of capsule
      -combination of Negative Stain and Simple Stain

25

Acid-Fast Stain

PRIMARY STAIN:
      -Carbolfuchsin + Phenol
MORDANT:
      -Heat/Steam
DECOLORIZER:
      -Acid Alcohol
SECONDARY STAIN:
      -Methylene Blue
RESULTS:
      -Acid-fast bacteria appear red
      -Non acid-fast bacteria appear blue
PURPOSE & COMMENTS:
      -to detect the presence of mycolic acid
      -differential staining technique

26

Gram Stain

PRIMARY STAIN:
     -Crystal Violet
MORDANT:
     -Gram's Iodine
DECOLORIZER:
     -95% Ethanol
SECONDARY STAIN:
     -Safranin
RESULTS:
     -Gram positive bacteria appear purple
     -Gram negative bacteria appear red
PURPOSE & COMMENTS;
     -Most common differential stain
     -to detect high peptidoglycan cell wall composition (Gram +: =90%, Gram -: =10%)

27

Spore Stain
(Schaefer-Fulton method)

PRIMARY STAIN:
     -Malchite Green
MORDANT:
     -Heat/Steam
SECONDARY STAIN:
     -Safranin
RESULTS:
     -Red cells: no spores
     -Red cells with green spots inside: spore forming bacteria
PURPOSE & COMMENTS:
     -differential stain
     -to detect the presence & location of spores
     -all endospore formers are typically Gram positive

28

Flagellar Stain
(Leifson's method)

PRIMARY STAIN:
      -Pararosaniline
MORDANT:
      -Tannic Acid
RESULTS:
      -observe flagella location and distribution
      -Amphitrichous, Monotrichous, Lophotrichous, Peritrichous
PURPOSE & COMMENTS:
      -difficult to do
      -detect the presence & arrangement of flagella
      -primary stain & mordant are collectively called Leifson's stain