Lab 5

Alistair WressniggBIOL 260 Fall 2011

Susanna HeinzeLab 5

Gram Stain Unknowns

1

#1Gram (–) Single rodsNo apparent errors.Does not rule out. All given bacteria have Gram negative single rods.

#2Gram (–) Single rodsNo apparent errors.Does not rule out. All given bacteria have Gram negative single rods.cocci





Susanna HeinzeLab 5

EMB

2

#1 – Good Growth No apparent errors

Rules out E.coli This bacteria shows Green/Brown growth.

#2 – Green/Black Growth No apparent errorsRules out E.aerogenes, K.pneumoniae, P.hauseri, P.aeruginosaDo not have G/B growth.

#3 – Pinkish Growth No apparent errors


#4 – Slightly Pinkish Growth No apparent errors


#5 – Pink Growth No apparent errors



Susanna HeinzeLab 5

SIM

3

#1 - No reaction: No positivesNo apparent errors

Rules out E.coli, E.aerogenes, P.haurseri, P.aeruginosa (have positive reactions)

#2 – Red, growth: Indole, MotilityNo apparent errors

Rules out K.pneumoniae (no reaction), E.aerogenes (no indole), P.hauseri, P.aeruginosa (sulfer pos)

#3 – Red, black prec: Sulfer ,Indole No apparent errors

Rules out K.pneumoniae (no reaction), E.aerogenes (no indole), P.hauseri, (sulfer pos)

#4 – Growth only: MotilityNo apparent errorsRules out K.pneumoniae (no reaction), E.coli, E.aerogenes, P.hauseri, P.aeruginosa (more reactions)

#5 – Black prec, red, growth Indole,Sulfer, Molility No apparent errorsRules out K.pneumoniae (no react), P.aeruginosa (no growth) E.coli, E.aerogenes,(no sulfer)

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Susanna HeinzeLab 5

O/F Glucose

4

#1 – Y/Y:Facultative anaerobe

No apparent errors

Rules out P.aeruginosa(G/G, Obligate aerobe)

#2 – Y/Y: Facultative anaerobe

No apparent errors


#3 – G/G: Obligate aerobe

No apparent errors

Rules out all but P.aeruginosa(Facultative anaerobes)

#4 – Y/Y: Facultative anaerobe

No apparent errors


#5 – Y/Y: Weak Facultative anaerobe

No apparent errors


Min

eral

Oil


Susanna HeinzeLab 5

Urease

5

#1 – Bright pink, growth: Positive

No apparent errors

Rules out E.coli, E.aerogenes (urease neg)

#2 - Yellow: Negative

No apparent errors

Rules out K.pneumoniae, P.hauseri, P.aeruginosa (urease pos)

#3 – Light pink, clear: Weak Positive

No apparent errors


#4 - Yellow: Negative

No apparent errors

Rules out K.pneumoniae, P.hauseri, P.aeruginosa (urease pos)

#5 – Bright pink, clear: Positive

No apparent errors



Susanna HeinzeLab 5

TSI

6

#1 – Yellow, gaps: GlucoseLactoseSucroseGasNo apparent errorsRules out P.aeruginosa(has sulfer), P.hauseri (has sulfer and glucose only)

#2 – Yellow, gaps: GlucoseLactoseSucroseGasNo apparent errorsRules out P.aeruginosa(has sulfer), P.hauseri (has sulfer and glucose only)

#3 – Red, black prec: GlucoseSulferNo apparent errorsRules out E.coli, E.aerogenes, K.pneumoniae (does not have sulfer production)

#4 – Yellow, gaps, pink t: Glucose LactoseSucroseGasNo apparent errorsRules out P.aeruginosa(has sulfer), P.hauseri (has sulfer and glucose only)

#5 – Yellow, cracks, black prec: SulferGLSGasNo apparent errorsRules out E.coli, E.aerogenes, K.pneumoniae (does not have sulfer production)


Susanna HeinzeLab 5

MR-VP

7

#1 - No reaction: MR+VP-

No apparent errors

Rules out E.aerogenes (VP+) P.aeruginosa (MR-)


No apparent errors


#3 - No reaction: MR-VP-

No apparent errors

Rules out E.aerogenes (VP+), E.coli, K.pneumoniae, P.hauseri, (MR+)

#4 - No reaction: MR-VP+

No apparent errors

Rules out E.coli, K.pneumoniae, P.hauseri, P.aeruginosa (VP-)


No apparent errors


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Susanna HeinzeLab 5

Lab 5 Conclusions & Sources of Error

In BIOL260 week five, Lab 5 covered tests to identify gram-negative rods.

The techniques I learned in this lab include demonstrating and completing the SIM,

O/F Glucose, Urease, TSI, and MR-VP tests. Please see the Dichotomous Key on the

next page.

8

#1 Klebsiella pneumonia: Gram(-) Rods, EMB+, SIM-, Facultative anaerobe, MR+, VP-, Urease+, Glucose+, Lactose+, Sucrose+, Gas

#2 Escherichia coli: Gram(-) Rods, EMB+, IM+, Facultative anaerobe, MR+, VP-, Urease-, Glucose+, Lactose+, Sucrose+, Gas

#3 Pseudomonas aeruginosa: Gram(-) Rods, EMB+, SI+, Obligate aerobe, MR-, VP-, Urease+, Glucose+, Sulfer+

#4 Enterobacter aerogenes: Gram(-) Rods, EMB+, Motility, Facultative anaerobe, MR-, VP+, Urease-, Glucose+, Lactose+, Sucrose+, Gas

#5 Proteus hauseri: Gram(-) Rods, EMB+, SIM+, Weak Facultative anaerobe, MR+, VP-, Urease+, Glucose+, Lactose+, Sucrose+, Gas

#1 Only SIM that had no result and all other results match.

#2 EMB had a green/black sheen seen with E.coli and all other results match.

#3 Only SIM with Sulfer and Indole but no growth, only Obligate aerobe, and all other results match.

#4 Only VP positive unknown with a pink top TSI; All other results match.

#5 Only SIM with Sulfer, Indole and growth, and all other results match.


Susanna HeinzeLab 5

9

Escherichia coli

EMB confirmed

Escherichia coli

MR-VP+

MR+VP-

Indole +Sulfer +Motility

Sulfer –Indole –

MR-VP

SIM

E.coliE.aerogenes

K.pneumoniaP.Hauseri

Gram (-) Rods

Escherichia coliEnterobacter aerogenes

Klebsiella pneumoniaProteus Hauseri

Pseudomonas aeruginosa

E.coliE.aerogenes

K.pneumoniaP.Hauseri

O/F Glucose

Urease

OF

Pseudomonas aeruginosa(OXIDIZER)

Proteus Hauseri

(-) (+)



Documents

Lab 5