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Laboratory 7 & 8 Bacteriology - PowerPoint PPT Presentation


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Laboratory 7 & 8 Bacteriology. Bacteria. Small Unicellular Organisms Can be grown in nutrient enriched environments (Agar, Broth) Standard Medias: Tryptic Soy Agar (TSA), Brain Heart Infusion (BHI), Tryptic Yeast Extract and Salt (TYES).

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bacteria
Bacteria
  • Small Unicellular Organisms
  • Can be grown in nutrient enriched environments (Agar, Broth)
  • Standard Medias:Tryptic Soy Agar (TSA), Brain Heart Infusion (BHI), Tryptic Yeast Extract and Salt (TYES)
common bacterial fish pathogens
Flavobacterium psychrophilum-Coldwater Disease-Gram Negative Rods

Renibacterium salmoninarum-Bacterial Kidney Disease-Gram Positive Rod

Yersinia Ruckeri-Enteric Red Mouth Disease-Gram Negative Rods

Aeromonas salmonicida-Furunculosis-Gram Negative Rod

Streptococcus iniae-O –sherical shape-Gram Postive

Common Bacterial Fish Pathogens
aseptic technique
Aseptic Technique
  • Minimizes contamination from outside sources
  • Prevents work surfaces from becoming contaminated
bacterial isolation
Bacterial Isolation
  • Streaking technique used to isolate individual bacterial colonies.
  • Can be used to isolate colonies in pure or mixed cultures
wet mounts
Wet Mounts
  • Used to determine motility of organism
  • Can help in bacterial identification
  • Do not confuse with Brownian motion
bacterial morphology
Bacterial Morphology
  • Colony Morphology
    • Size
    • Shape
    • Color
bacterial morphology8
Bacterial Morphology
  • Cell Morphology
    • Cocci
    • Bacillus
    • Spirochete
interpreting antibiotic sensitivity test
Interpreting Antibiotic Sensitivity Test
  • Measure diameter of zone of inhibition (mm)
  • The larger the zone, the more effective an antibiotic will be.
gram staining

+ -

Gram Staining

Heat Fixed

  • Gram positive organisms retain the primary stain
  • During decolorization gram negative organisms will lose pigment and pick up the counter stain

Crystal Violet

Gram’s Iodine

Ethyl Alcohol

Safranin

gram staining11
Gram Staining

Disadvantages:

  • Some bacteria are Gram stain variable (positive or negative results)
  • Some bacteria are resistant to Gram stain (i.e. acid-fast bacteria)
  • False results may occur if over-decolorized
  • Older cultures may give false results
koh potassium hydroxide test
KOH (Potassium Hydroxide) Test
  • Alternative to Gram stain
  • Stringy and viscous = Gram negative
  • No viscous or stringy = Gram positive
  • KOH is caustic: wear gloves and immediately flush skin or eyes with water if they come into contact with KOH solution
cytochrome oxidase test
Cytochrome Oxidase Test
  • Identifies organisms that produce cytochrome oxidase
  • Solution remains colorless = negative (doesn’t produce cytochrome oxidase)
  • Solution turns purple or blue = positive (does produce cytochrome oxidase)
  • Results appear within 15 seconds