Lab Practical 2 Flashcards
All GN Cocci
key characteristic
Neisseria gonorrhoeae
Neisseria meningitidis
Neisseria lactamica
Moraxella catarrhalis
kidney bean shaped
All GN dipplococci and fastidious GN bacilli
Haemophilus influenzae
Haemophilus parainfluenzae
Brucella
Francisella tularensis
Legionella
All Enterobacterales
Citrobacter freundii
Enterbacter cloacae
Escherichia coli
Klebsiella pnemoniae
Proteus mirabilus
Vibrio
Shigella
Salmonella
E. coli O157
All Remaining MISC GNR
Acinetobacter baumannii (ACBA)
Burkholderia cepacia (BUCE)
Stenotrophomonas maltophilia (STMA)
Pseudomonas aeruginosa (PSAE)
Yersinia
MAC Agar
type, additives, growth indicators
Slective differential
Selective: bile salt + crystal violet, selects for G-
Differential: lactose fermentation > turns agar pink
Growth: enterobacterales mostly
All Mediums for Neisseria
3 type, reagents
Thayer Martin, Modified Thayer Martin, Martin Lewis
All are modified CHOC with vancomycin
What Agar is Used to Isolate Bordatella?
2 name, type, reagents
Bordet-gengou (B-G) blood agar
- Enriched + selective
- Blood and glycerol for enrichment, antimicrobials for selectivity
- For B. pertussis and parapertussis
Regan-Lowe agar
- B. pertussis
- Charcoal
Legionella Isolating Medium
Name, type, how
BCYE-alpha (buffered charcoal-yeast extract) Agar
- Enriched
- L-cysteine added which Legionella needs
- Charcoal absorbes toxic compounds
SMAC Agar
type, how, what for
Selective differential
- Sorbitol replaces lactose
- Normal E. coli flora can ferment sorbitol, E coli. O157 is colorless
HE Agar
what species, reagents,
Salmonella and Shigella
- Lactose, sucrose, bile salt, ferric ammonium sulfate for H2S
- Salmonella produces H2S, Shigella does not
CIN Agar
2 species, reagent
Isolation of Yersinia, sometimes Citrobacter can grow
- Yersinia appears translucent without dark red centers
- Novobiocin antimicrobial
Medium for Vibrio Isolation
Thiosulfate Citrate Bile salts Sucrose (TCBS) agar
- Thiosulfate
- oxgall
- sucrose use = yellow color
Neisseria + Moraxella Lactose
Neisseria gonorrhoea: -
Neisseria lactamica: +
Moraxella: -
Neisseria meningitidis: -
Neisseria + Moraxella Glucose
All Neisserias can use
Moraxella cannot use
Neisseria + Moraxella Maltose
Lactamica and meningitidis can
Gonorrhoea and Moraxella cannot
Butyrate test
method and who is +
Production of butyrate esterase causes release in indoxyl. Forms indigo, blue green color change
Only Moraxella catarrhalis is +
Oxidase
method
Kovac reagent oxidized by electrons, makes purple color
Porphyrin
method, what for, results
Porphobilinogen presence, ALA will convert to porphyrin, fluorescent
- Separate H. influenzae and parainfluenzae
- Tests for hemin
+: parainfluenzae
-: influenzae
Satellite Growth
H. influenzae needs both NAD (V) and hemin (X) that it will grow around S. aureus bc it lyses the blood and releases V factor
Oxidation Fermentation (O/F)
how, what species and results
How: sugar and pH indicator, oil layer for anaerobic
Fermenter: E. coli
Oxidizer: Pseudomonas aeruginosa
Pigment Production of Pseudomonas Aeruginosa
blue and fluorescent coloring
Spot Indole
reagents, interpretation
Trypyophanase enzyme detection, degrade indole because of degradation of tryptophan
- Red color +
Separate E. coli from other enterbacterales becaue E.coli is +
ONPG Test
method, results
B- galactosidase cleaves ONPG into galactose
- Positive if broth turns yellow
E.coli is + while Proteus mirabilis is -
MUG test
method, results
Tests for beta glucuronidase, fluorescence
Only E. coli is +
TSI Test
method, results
Glucose, sucrose, lactose, ferric and sodium thiosulfate
A/A: glucose + something else
K/A: only glucose
K/K: none of the sugars
H2S: has to be either A/A or K/A because enterbacterales always uses glucose, only ones that can produce H2S
Motility Test
Nonmotile enterobacterales: Klebsiella pnuemoniae and Shigella
Nonmotile Misc GNR: acinetobacter baumannii
Citrate Agar
Can utilize citrate, slant turns blue
Klebsiella pneumoniae +
E. colli -
Good Test to Differentiate Proteus Mirabilis
But also what?
Urea hydrolysis
- CIFR, ENCL, and KLPN and also split urea into ammonia
MR/VP
2 reagents in VP, enterobacterales
VP: KOH and alpha-naphthol
Positive for VP: ENCL and KLPN
- Everything else negative
- Vice versa for MR: ENCL and KLPN are negative for MR while everything else +
Nitrate Reduction
what kinds of reduction? 2 reagents in broth
All enterobacterales can reduce nitrate
- Can go directly to N2 gas or
- alpha napthylamine and sulfanilic acid for reduction to nitrite
- Turns red if nitrite
- If colorness, add zinc and red color means there is still nitrate, no color means N2 produced and no nitrate left
Yersinia Bench Aid
- Catalase +
- Nonmotile at 25 and 35/37
- Urease, Oxidase, Indole -
- ## Bipolar staining safety pin appearance
Burkholderia Bench Aid
B. mallei: oxidase -, non motile
B. pseudomallei: opposite of that
Francisella Bench Aid
- No SBA growth
- Oxidase, urease -
- Catalase weak +
Brucella Bench Aid
SBA and CHOC
- Oxidase, urease +
- Non motile
Oxidase + Rods
2, differentiate
Burkholderia cepecia
Pseudomonas aerogunosa
- Then do colony morph, pseudomonas smell and sheen
Nonmotile Rods
3, differentiate
Klebsiella pneumoniae
Shigella
Acinetobacter baumannii
- MAC lactose, ACBA is non fermenter
Motile Rods
5, how to differentiate
ESCO
ENCL
STMA
PRMI
CIFR
- Then do tsi h2s production
TSI H2S Production in Rods
+ : PRMI, CIFR
- : ENCL, ESCO, STMA
Differentiate ESCO and ENCL
Indole
- E. coli is +
- Enterobacter cloacae is -
Differentiate CIFR and PRMI
Proteus mirabilis swarms on SBA, Citrobacter does not
