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Basic Dye
Methylene Blue, Safranin, Crystal Violet
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Acidic Dye
What does it stain?
Eosin, Congo Red, Nigrosine
Used to stain Background
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Gram Stain
Order?
- Crystal Violet
- Iodine,
- Alcohol
- Safranin
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Capsul Stain
Stains bacteria with capsules
Use Congo Red or Crystal Violet
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Endospore Stain
What organism does it test for?
Stains spores with Malachite Green
Tests for Bacillus Clostridium
Spores are green, vegetative cells are pink
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Motility Test
What Media?
Organisms?
Mot-S or Sim tube
Flagellates or other motile organisms
Cloudiness is a positive test.
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Sulfur Utilization?
Media?
Organisms?
- Black is positive result
- SIM Tube or KIA
- Proteus Vulgaris
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Aerobe vs Anaerobe?
Media?
Use a Thioglycolate tube.
Aerobes on surface, anaerobes grow through medium. Faculitive Anaerobes near surface
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Antimicrobial/ Antibiotic Sensitivity
Medium?
Kirby Bauer Method on Mueller Hinton Agar plate
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Oxidase
Organisms?
Medium?
Positive Result?
- + Neisseria
- + Pseudomonas
- - Enterobacteria
- TSA Plate
- Pink, Maroon color. Turns blue after 60 sec
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Phenylalanine Deamination
Organisms?
Medium
Positive Result
- Proteus
- Providentia
- Morganella
Slant Tube
Green on slant
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Staphylococci Identification
Tests
- Catalase: Staph Aureus is +
- MSA: Yellow= aureus
- Blood Agar: Clear Zone is Aureus
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Streptococci Identification
Tests
- Blood Agar:
- Bacitracin A=pyogenes, CAMP=agalactiae, P=Pneumonia
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Hemolysis
Beta
Alpha
Gamma
- Beta: Clear zone
- Alpha: Green Zone
- Gamma: No clear zone
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Starch Hydrolysis
Organism
Medium
- Bacillus subtilis
- Starch agar plate
- Clear zones is positive result
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Tryptophan
Organism
Media
Reagent
Positive Result
- Citrobacter, some Shigella
- SIM
- Kovac's Reagent
- Red Layer on top
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Urea Hydrolysis
Organism
Media
Positive
- Proteus Providencia Morganella
- Urea Broth (Phenol Red in media)
- Red color
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Yogurt Production
Organism
Strep Thermophilus
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Lactose Positive Organisms
- Escheracia
- Enterobacter
- Klebsia
- Citrobacter
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Total Magnification
Ocular x Objective
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Limit of Resolution
Minimum distance between two objects that are discernable to see them both
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Acid Stains have what charge?
Basic Stains have what charge?
Bacterial Cell walls have what charge?
- Negative Charge
- Positive Charge
- Negative Charge
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Differential Stain Purpose
Example
- Two different stains
- Gram stain or Acid fast stain
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Flagella
Monotrichous
Lophotrichous
Amphitrichous
Peritrichous
- M: Single Flagella
- L: Many on one side
- A: One on each side
- P: Many all over
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Catalase
Organisms
Results
- + Staphylococcus
- - Enterococcus
- -Strep
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Enterics
Gram?
Oxidase
Glucose
Nitrite
Examples
- Gram negative
- Oxidase Negative
- Glucose Positive
- Nitrate Positive
- Examples: E. Coli, Klebsia, Enterobacter, Salmonella, Shigella
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Psuedemonas
Oxidase
Motility
Fermentation
- Oxidase Positive
- Motile
- Fermentation Negative
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Differential Media
Examples
Some metabolic characteristic of an organism is detected due to media
KIA, MaConkie Agar, Urea, Starch Hydrolysis
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Selective Media
Allows the growth of certain organisms and inhibits others
MSA, Bile Eschulen
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Enriched Media
Rich in nutrients
Blood Agar, Chocolate Agar,
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Lactose Fermenter
All Coliforms
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Countable Plates
20-80 coliform colonies
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Protozoa
- Amoebas:Entamoeba
- Ciliates: Balantidium
- Flagellates: Giardia, Trichomonas
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Platyhelminthes
- Fasciola hepatica
- Shistosoma
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Bacillus
Gram
Morphology
Media
Pos Results
- Positive
- Rod
- TSA Catalase +
- Starch Digestion
- H2O2 and Gas Bubbles
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Citrobacter
Gram
Morphology
Media
Positive Results
- Gram Negative
- Morphology: Rod
- Media:
- MAC- Red or pink colonies
- EMB- same as above
- KIA -A/A++
- H2S +
- Lactose+
- Glucose +
- Oxidase -
Urea-No growth
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Clostridium
Gram
Morphology
Media
Positive Results
- Gram Positive
- Morphology Rod
- Media:
- TSA-growth
- Blood Agar-hemolysis
- Catalase -
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Micrococcus luteus
Gram
Morphology
Media
- Gram Positive
- coccus
- TSA-Yellow colonies
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Neiseria
Gram
Morphology
Media
- Gram Negative
- Diplococcus
- Thayer Martin
- Chocolate Agar
- Oxidase positive
- purple color change
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Proteus Vulgaris
Gram
Morphology
Media
- Gram Negative
- Rod
- MAC/EMB-colorless
- KIA -black
- Urea Red
- Lactose -
- Glucose +
- Oxidase -
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Pseudomonas
Gram
Morphology
Media
- Gram Negative Rod
- KIA-Alk/Alk. - -
- TSA -green pigment
- Glucose -
- oxidase +
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Salmonella
Gram
Morphology
Media
- Gram Negative rod
- KIA-Alk/Acid +- Black
- Urea -
- Lactose -
- Glucose +
- Oxidase -
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Serratia
Gram
Morphology
Media
- Gram Negative Rod
- KIA-Acid/Acid --
- TSA 25 degree -red pigment
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Staph Aureus
Gram
Morphology
Media
- Gram Positive Rod
- Blood Agar: Beta Hemolysis
- MSA: Yellow media
- Catalase +
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Staph epidermis
Gram
Morphology
Media
- Gram Positive Coccus
- Blood Agar: Gamma Hemolysis
- MSA: Pink media and colonies
- Catalase +
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Strep feacalis
Gram
Morphology
Media
- Gram Positive Coccus
- Blood agar: alpha hemolysis
- Bile Eschulen: more than 1/2 slant is black +
- catalase -
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Strep Pyogenes
Gram
Morphology
Media
- Gram positive Coccus
- Blood agar: beta hemolysis, bacitracin sensitive
- Bile Esculin -
- catalase-
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E. Coli
Gram
Morphology
Media
- Gram Negative Rod
- EMB: metallic green
- MAC: pinka
- H2S:-
- Lactose +
- Glucose+
- Oxidase -
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Enterobacter
Gram
Morphology
Media
- Gram Negative Rod
- EMB: Color
- Mac: Color
- Motility+
- Indole -
- Lactose +
- Glucose +
- Oxidase -
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Klebsia
Gram
Morphology
Media
- Gram Negative Rod
- EMB: Color
- MAC: Mucoid
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Shigella
Gram
Morphology
Media
- Gram Negative Rod
- EMB: Colorless
- MAC Colorless
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