9
Gram Stain + Blue Morphology Cocci Catalase Test + Bubbles BAP Beta hemolysis (double zone) Staphylococcus aureus Confirm: MSA - Yellow Gamma hemolysis Staphylococcus epidermidis Confirm: MSA - Pink - No Bubbles BAP Gamma hemolysis Enterococcus faecalis Confirm: TSA Small white colonies Beta hemolysis (single zone) Streptococcus pyogenes Confirm: TSA - Tiny pinpoint white colonies Bacilli Bacillus subtilis Confirm: BAP – Alpha hemolysis BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn

BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

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Page 1: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

Gram Stain + Blue

Morphology

Cocci

Catalase Test

+ Bubbles

BAP

Beta hemolysis (double zone)

Staphylococcus aureus

Confirm: MSA - Yellow

Gamma hemolysis

Staphylococcus epidermidis

Confirm: MSA - Pink

- No Bubbles

BAP

Gamma hemolysis

Enterococcus faecalis

Confirm: TSA Small white

colonies

Beta hemolysis (single zone)

Streptococcus pyogenes

Confirm: TSA - Tiny pinpoint white colonies

Bacilli

Bacillus subtilis

Confirm: BAP – Alpha hemolysis

BIO 260L – Microbiology Gram+ Dichotomous Key

SP 14 - Fairborn

Page 2: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

Gram Stain - Red

Green pigmentation on

TSA (48h)

- Not green

Lactose Test

+ Yellow

Urease Test

+ Pink (delayed)

Klebsiella pneumoniae

Confirm: SIM: -Indole No red ring

- Yellow

Escherichia coli

Confirm: SIM:+Indole, red

ring

- Pink

SIM Test: Indole

+ Red ring

Proteus vulgaris

Confirm: SIM: Moderate Motility

- No red ring

Proteus mirabilis

Confirm: SIM: Highly Motile

+ Green

Pseudomonas aeruginosa

Confirm: Oxidase+ Blue

BIO 260L – Microbiology Gram- Dichotomous Key

SP 14

Page 3: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

Gram Stain + Blue

Morphology

Cocci

Catalase Test

+ Bubbles

BAP

Beta hemeolysis (double zone)

Staphylococcus aureus

Confirm: MSA - Yellow

Gamma hemeolysis

Staphylococcus epidermidis

Confirm: MSA - Pink

- No Bubbles

BAP

Alpha hemolysis

Streptococcus pneumoniae

Confirm: TSA Small white

colonies

Beta hemolysis (single zone)

Streptococcus pyogenes

Confirm: TSA - Tiny pinpoint

colonies

Bacilli

Bacillus subtilis

Confirm: BAP – Alpha

hemeolysis

BIO 260L – Microbiology Gram+ Dichotomous Key

WI 13

Page 4: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

Gram + (Blue- purple) Morphology

Coccus Rods

Staphylococcus aureus Staphylococcus epidermidis

Enterococcu faecalis Streptococcus pyogens Micrococcus leuteus

Bacillus subtilis

Catalase (H2O2) + Bubbles - Bubbles

S. aureus S. epidermidis Micrococcus

Enterococcus S. pyognes

BAP : Alpha hemolysis

TSA-Pigment

Off white Yellow/Orange

S. aureus S. epidermidis

Micrococcus

Confirm: BAP Gama hemolytic

MSA Pink Yellow

S. aureus

Confirm: Double zone of

Beta hemolysis

S. epidermidis

Alpha/Beta hemolysis

BAP Alpha

hemolytic Beta

hemolytic

Enterococcus S. pyogenes

Confirm: TSA Tiny Pinpoint Colonies

Confirm: TSA Small Off White

Colonies

Page 5: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

Gram – (Pink – Red)

Lactose + Yellow - Pink

Pseudomonas aeurginosa Serratia marcescens

Proteus vulgasris Proteus mirabilis

SIM - Sulfur +

Black ppt

- No Black ppt

Proteus vulgaris Proteus mirabilis

SIM -Indole + red ring - No red ring Proteus vulgaris

Confirm: SIM

Slightly motile

Proteus mirabilis

Confirm: SIM

Highly motile

Pseudomonas Serrita

Oxidase + -

Pseudomonas Serratia

Confirm: Green on TSA

48hrs

Confirm: Pink on TSA

25 C

E. coli Confirm: citrate

(blue)

Page 6: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

Gram – (Pink Red) Oxidase    

Blue + -

White

Pseudomonas Confirm: Green on

TSA 48hrs

E. coli Klebsiella

Serrtia Proteus vulgaris Proteus mirablils

TSA pigmentation

White    - Yellow

+

E.coli Klebisella

Serrita

Proteus vulgaris Proteus mirabilis

Lactose +    Yellow

- Pink

E. Coli Confirm: Citrate

Green

Serratia Confirm: TSA 25C

SIM - Motility Motile Highly Motile

Proteus vulgaris Confirm: Indole

+ red ring

Proteus mirabilis Confirm: Indole – No red ring

Klebisella Confirm:

Citrate + Blue

Page 7: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

GRAM  STAIN  

Gram Negative

Rod

Oxidase  TEST  

Lactose  ferm  on  MAC    

H₂S  prod  in  SIM  

+  

P.  vulgaris  

+  

E.  coli  

+  

P.  aeruginosa  

Cocci

Gram Positive

Rod Cocci

Catalase  TEST  

Hemolysis  TEST  

Alpha  

Optochin  InhibiDon  

S.  pneumoniae  

Beta  

S.  pyogenes  

+  

Mannitol  plate  /  acid  

S.  epidermidis  

+  

S.  aureus  

Page 8: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology
Page 9: BIO 260L – Microbiology Gram+ Dichotomous Key Gram … · BIO 260L – Microbiology Gram+ Dichotomous Key SP 14 - Fairborn . ... Enterococcu faecalis Streptococcus pyogens ... Microbiology

Gram Stain +blue -red

S. aureus S. epidemidis

E. faecalis S. pyogens

E. coli Salmonella P. mirabilis

Pseudomonas

Catalase Mackonkey’s + bubbles - bubbles

S. aureus S. epidermidis

E. faecalis S. pyogens

Red Colorless

E. coli Pseudomonas Salmonella

P. mirabilis

MSA Pink Yellow

Staphyloccous epidermidis

Confirm: Coagulase –

BAP: Gamma-hemolytic

Staphyloccous aureus

Confirm: Coagulase + BAP: Double zone of Beta-

hemolysis

Bacitracin Resistant Inhibited

Enteroccous faecalis

Confirm: BAP: Gamma-

hemolysis

Streptococcous pyogens

Confirm: BAP: Beta-hemolysis

TSI A A

Gas +

K K Gas -

Escherichia coli

Confirm: TSA: white

pigmentation

Pseudomonas aeruginosa

Confirm: TSA: green

pigmentation after 48hrs

TSI K A Bppt

K Bppt

Salmonella enteritidis Confirm:

TSA: white pigmentation

Proteus mirabilis

Confirm: TSA: yellow

pigmentation

BIO 260L – Microbiology Lab 8 and 9 Dichotomous Key for Lab Exercises 45, 46, 48 Bacterial Diseases of the Skin, Respiratory, and GI Systems