Scc-Mlt-Microbiology 2, Unit 1

Front 1

Tribe I

Escherichieae

Back 1

Escherichia coli

Escherichia coli 0157: H7

Shigella boydii

Shigella dysenteriae

Shigella flexnerii

Shigella sonnei

Front 2

Tribe II

Edwardsielleae

Back 2

Edwardsiella

Front 3

Tribe III

Salmonelleae

Back 3

Salmonella typhimurium

Front 4

Tribe IV

Citrobactereae

Back 4

Citrobacter freundii

Front 5

Tribe V

Klebsielleae

Back 5

Enterobacter aerogenes

Enterobacter cloacae

Klebsiella oxytoca

Klebsiella pneumoniae

Serratia marcescens

Hafnia species

Front 6

Tribe VI

Proteeae

Back 6

Proteus mirabilis

Proteus vulgaris

Morganella morganii

Providencia stuartii

Front 7

Tribe VII

Yersinieae

Back 7

Yersinia

Front 8

Enterics virulence factors

Back 8

- produce endotoxins

- often antibiotic resistant

- colonize or invade tissues (invasive)

Front 9

Enterobacteriaceae characteristics

"Enterics"

Back 9

- Gram negative rods (short & chubby)

- All ferment glucose

- All reduce nitrate to nitrite

- Motile (except Klebsiella, Shigella and Yersinia)

- Non fastidious (dont need special nutrient media to grow)

Front 10

MAC

MacConkey agar

Back 10

- Mild to moderately selective (grows gram -)

- Differential for lactose fermenters

- Lactose ferment = pink

- no lactose ferment = colorless

pH indicator = neutral red

Bile salt/crystal violet = suppresses gram +

Good for Enteric stool pathogen/urine culture

Front 11

EMB

Eosin methylene blue

Back 11

- Mild to moderately selective (grows gram -)

- Differential for lactose fermenters

- Lactose ferment = metallic green (E. coli) or opaque purple

- no lactose ferment = colorless

methylene blue/eosin = suppresses gram +

Front 12

HE

Hektoen enteric agar

Back 12

- Moderate to highly selective (grows gram -)

- Differential for lactose/sucrose fermenters

---common use to diff. Salmonella/Shigella

- Lactose/sucrose ferment = salmon/orange

- non fermenters of these = clear green

- H2S = black dot

- Bile salts = suppresses gram + and nonpathogenic Enterics

- sodium thiosulfate/ferric sulfate = H2S producers cause black dot

Front 13

XLD

Xylose Lysine Desoxycholate agar

Back 13

- Moderate to highly selective (grows gram -)

---less selective than HE or S-S media

- Differential for lactose/sucrose/xylose fermenters

---good media to isolate Shigella from enrichment broth

- Lactose/sucrose/xylose ferment = yellow

- non fermenters of these = clear red

- H2S = black dot

- sodium thiosulfate/ferric sulfate = H2S producers cause black dot

Front 14

S-S

Salmonella-Shigella agar

Back 14

- Moderate to highly selective (grows gram -)

- Differential for lactose fermenters

---common use to diff. Salmonella/Shigella

- Lactose ferment = pink

- non fermenters of these = colorless

- H2S = black dot

- Bile salts/sodium citrate = suppresses gram + and nonpathogenic Enterics

- sodium thiosulfate/ferric sulfate = H2S producers cause black dot

Front 15

MAC-S

MacConkey with sorbital agar

Back 15

- Mild to moderately selective (grows gram -)

- Differential for sorbital fermenters

- Sorbital ferment = pink

- no sorbital ferment = colorless

---used to identify E. coli 0157:H7 (negative)

pH indicator = neutral red

Bile salt/crystal violet = suppresses gram +

Good for Enteric stool pathogen/urine culture

Front 16

BG

Brilliant Green agar

Back 16

Highly selective media

--only grows Salmonella

Very expensive, not used in labs often

Front 17

Selenite broth

(for pathogenic Enterics)

Back 17

Isolate Salmonella from feces, urine or sewage

Enriched broth

- Contains lactose and peptones

Front 18

Gram Negative broth

(for pathogenic Enterics)

Back 18

Inhibits gram positive growth

- may inhibit Proteus

- increase isolation of Shigella/Salmonella

Enriched broth

- contains glucose, mannitol, deoxycholate

Front 19

KIA

Kligler Iron agar tube

sugar fermentation examples

Back 19

Top slant = lactose

Bottom slant = glucose

1-no sugars fermented (K/K)

2-glu and lac (A/A)

3-glu and lac and gas (A/A)

4-only glu (K/A)

5-only glu and H2S (K/A)

**K=alkaline A=acid

Front 20

TSI

Triple Sugar Iron agar tube

sugar fermentation examples

Back 20

Top slant = lactose &/or sucrose

Bottom slant = glucose

Front 21

MR-VP broth

Methyl Red / Voges-Proskauer test

glucose metabolism examples

Back 21

A = MR negative

B = MR positive

C = VP positive

D = VP negative

Front 22

Citrate slant

Citrate utilization test example

Back 22

A = negative

B = positive

Salmonella vs C. freundii

Providencia vs Morganella

Front 23

Indole production test

Back 23

E. coli = positive

Indole is the reason for E. coli's glue smell

Reagents - Kovac's or Ehrlich's

primary ingredient = 10% para-dimethylaminobenzaldehyde

K. pneumonia vs K. oxytoca

P. vulgaris vs P. mirabilis

Front 24

Gelatin test

Back 24

Bacteria with enzyme gelatinase, can break down gelatin into a liquid

Serratia = positive

Front 25

Malonate utilization test

Back 25

- Organisms that use malonate as source of carbon will grow

- Malonate utilization = blue (due to ammonia salts)

- Sodium hydroxide NaOH is the end product

**Klebsiella = positive (blue)

Front 26

Motility tests

SIM (sulfide, indole, motility)

MIO (motility, indole, ornithine)

Back 26

SIM test - room temp

Left = non motile

Right = motile

**Shigella and Klebsiella are non-motile

Front 27

Urease

Christensen's urea slant

Back 27

Pink = positive

Peach = negative

Organisms with enzyme urease will hydrolyze urea, releasing ammonia (an alkaline end product, causing pink color)

**Proteus = positive

Front 28

ONPG

Ortho-nitrophenyl-B-D-galactopyranoside

Back 28

C. freundii vs Proteus spp

C. freundii vs Salmonella

S. sonnei vs S. flexneri

Front 29

Escherichia coli

Back 29

Lactose +

Sucrose +

Sorbitol +

IMViC + + - -

Positive tubes:

LDC, Motile, Ind, ODC, MR, ONPG

*normal GI flora

*pathogenic when in cut or wound

*ESBL- Extended Spectrum Beta Lactamase producer (resistant to penicillin)

*Dysentery like diarrheal disease, UTI, endocarditis

Front 30

Escherichia coli 0157:H7

Back 30

Lactose +

Sucrose +

Sorbitol -

​IMViC + + - -

Positive tubes:

LDC, Motile, Ind, ODC, MR, ONPG

*Produces shiga-toxins

-Verotoxin I and II, Enterohemolysin (bloody diarrhea, kidney failure)

Toxic Categories:

**Enterohemorrhagic (EHEC) -associated with hemolytic uremic syndrome. Can cause hemorrhagic colitis.

-Enterotoxigenic (ETEC), Enteropathogenic (EPEC), and Enteroinvasive (EIEC)

Front 31

Citrobacter freundii

Back 31

Lactose +

Sucrose +

Sorbitol +

Positive tubes:

Motile, MR, Cit, Mal, H2S, ONPG

Front 32

Enterobacter aerogenes

Back 32

Lactose +

Sucrose +

Sorbitol +

Positive tubes:

LDC, Motile, ODC, VP, Cit, Mal, ONPG

Front 33

Enterobacter cloacae

Back 33

Lactose +

Sucrose +

Sorbitol +

Positive tubes:

Motile, ODC, VP, Cit, Mal, Urea, ONPG

Front 34

Enterobacter sakazakii

Back 34

Lactose +

Sucrose +

Sorbitol +

Front 35

Klebsiella pneumoniae

Back 35

Lactose +

Sucrose +

Sorbitol +

*ESBL- Extended Spectrum Beta Lactamase producer (resistant to penicillin)

Positive tubes:

LDC(or -), VP, Cit, Mal, Urea(weak), ONPG

Front 36

Klebsiella oxytoca

Back 36

Lactose +

Sucrose +

Sorbitol +

Positive tubes:

LDC, Ind, VP, Cit, Mal, Urea, ONPG

Front 37

Serratia marcescens

Back 37

Lactose -

Sucrose - (slow or variable)

Sorbitol +

Positive tubes:

LDC, Motile, ODC, MR, VP, Cit, ONPG

Front 38

Morganella morganii

Back 38

Lactose -

Sucrose -

Sorbitol -

Positive tubes:

Motile, Ind, ODC, MR, Urea, PDA

Front 39

Providencia stuartii

Back 39

Lactose -

Sucrose -

Sorbitol -

Positive tubes:

LDA, Motile, Ind, MR, Cit, PDA

Front 40

Proteus vulgaris

Back 40

Lactose -

Sucrose +

Sorbitol -

Swarms on BAP or CAP

Positive tubes:

Motile, Ind, MR, Urea, PDA, H2S

Front 41

Proteus mirabilis

Back 41

Lactose -

Sucrose -

Sorbitol -

Swarms on BAP or CAP

Positive tubes:

LDA, Motile, ODC, MR, Cit, Urea, PDA, H2S

Front 42

Salmonella typhimurium

Back 42

Lactose -

Sucrose -

Sorbitol +

Positive tubes:

LDC, Motile, MR, H2S

Front 43

Shigella boydii

Back 43

Lactose -

Sucrose -

Sorbitol + (only + Shigella)

Positive tubes:

MR

Front 44

Shigella dysenteriae

Back 44

Lactose -

Sucrose -

Sorbitol -

Positive tubes:

MR

Front 45

Shigella flexneri

Back 45

Lactose -

Sucrose -

Sorbitol -

Positive tubes:

MR

Front 46

Shigella sonnei

Back 46

Lactose -

Sucrose -

Sorbitol -

Positive tubes:

ODC, MR, ONPG

Front 47

Hafnia species

Back 47

Lactose -

Sucrose -

Sorbitol -

Positive tubes:

LDC, Motile, ODC, VP, Mal(or -), ONPG(or -)

Front 48

Shigella groups

Back 48

Shigella boydii - group C

Shigella dysenteriae - group A

Shigella flexneri - group B

Shigella sonnei - group D

To remember:

Shigella names in alphabetic order - groups spell out CABbeD.

**Shigella made me sick, so I CABbeD it home!!**

Front 49

Salmonella

Back 49

* Always pathogenic

*communicable and REPORTABLE

*2400 serotypes

- including all subspecies of Salmonella enterica

- we only work with Salmonella typhimurium

LDC + (diff from C. freundii)

Urea - (diff from Proteus)

Motile (diff from non-motile Shigella)

Isolate with MAC/EMB and HE/XLD/S-S

Front 50

IMViC

Back 50

I - Indole

M - Methyl Red

V - Voges Prauskauer

i

C - Citrate

Front 51

E. coli vs. Shigella spp

Back 51

E. coli vs. Shigella spp

LDC + -

Front 52

C. freundii vs. Proteus spp

Back 52

C. freundii vs. Proteus spp

PDA - +

ONPG + -

Front 53

Salmonella spp vs. C. freundii

Back 53

Salmonella spp vs. C. freundii

LDC + -

ONPG - +

KCN - +

Cit - +

Front 54

Providencia spp vs. Morganella spp

Back 54

Providencia spp vs. Morganella spp

LDA + -

ODC - +

Cit + -

Front 55

K. pneumonia vs. K. oxytoca

Back 55

K. pneumonia vs. K. oxytoca

Indole - +

Front 56

S. sonnei vs. S. flexneri

Back 56

S. sonnei vs. S. flexneri

ODC + -

ONPG + -

Front 57

P. vulgaris vs. P. mirabilis

Back 57

P. vulgaris vs. P. mirabilis

Indole + -

ODC - +

LDA - +