ENTEROBACTERIACEAE
- Often referred to as enterics 3. Hektoen Enteric Agar (HEA)
- Gram negative (-)
- Non-spore-forming
- Facultative anaerobe bacilli
- All are oxidase negative (-)
o Except for: Plesiomonas (oxidase
positive +)
- All are catalase positive (+)
- All are glucose fermenters (+)
- All are motile (+)
o Except for: SKY (Shigella, Klebsiella,
Yersinia)
o Klebsiella and Enterobacter – exhibits
mucoid colonies on plated media
o Proteus – swarming motility on
plated media
- EMBL MAC, HEA, XLD – media for
presumptive identification
SELECTIVE MEDIA FOR PRESUMPTIVE
IDENTIFICATION
1. MacConkey Agar
- Selects gram negative (-) bacteria
- Differentiates lactose fermenters (LF) and
non-lactose fermenters (NLF)
➢ Bile salts and crystal violet – inhibit gram
positive (+) bacteria
➢ Carbohydrate source: lactose
➢ Indicator: neutral red
➢ Lactose fermentation
o Positive = pink
o Negative = colorless
2. Eosin Methylene Blue Agar (EMB)
- Selects gram-negative bacteria
- Differentiates LFs from NLFs
➢ Eosin and methylene blue – inhibit gram
positive (+) bacteria
➢ Differentiation on Fermentation – lactose
and sucrose
➢ Lactose fermentation
o Positive = violet colonies
▪ E.coli = metallic green sheen
o Negative = colorless
- Inhibits normal flora coliforms
- Differentiates LFs from NLFs
➢ Bile salts – inhibit gram positive (+) bacteria
➢ Carbohydrate source: lactose, sucrose,
salicin
➢ Indicator: Bromthymol blue
➢ Sulfur source: Sodium thiosulfate
➢ Detect H2S Production: Ferric ammonium
citrate
o Blackening in the media
- Most non-pathogens ferment lactose and
sucrose
o Shows orange color (low pH)
o Green to orange – non-pathogenic
- Pathogens (Salmonella and Shigella)
o Green to blue green color with H2S
gas creating a black precipitate (no
fermentation)
4. Xylose-Lysine-Desoxycholate (XLD) Agar
- Selects for stool pathogens
- Differentiates xylose fermenters (yellow)
from non-xylose fermenters (red)
- Selective for Salmonella and Shigella from
stool specimens
➢ Sodium deoxycholate – inhibit gram positive
(+) cocci, gram negative (-) coliforms,
swarming of Proteus
➢ Detect decarboxylation – Lysine
➢ Carbohydrate source: Xylose
➢ Indicator: Phenol red
➢ Sulfur source: Sodium thiosulfate
➢ Detect H2S: Ferric ammonium citrate
5. Salmonella-Shigella Agar
- Selects for stool pathogens
- Differentiates LFs (pink-red) from NLFs (clear)
➢ Bile salts and brilliant green agar – inhibit
geam positive (+), gram negative (-) coliforms
➢ Carbohydrate source: lactose
➢ Indicator: neutral red
➢ Detect H2S: Ferric ammonium citrate
 MAC EMB HEA XLD S-S
Inhibit gram (+) Bile salts and Eosin and Bile salts Sodium Bile salts and
bacteria crystal violet methylene blue deoxycholate brilliant green
agar
Carbohydrate Lactose Lactose Lactose Xylose Lactose
source Sucrose Sucrose
Salicin

Indicator Neutral red Bromthymol Phenol red Neutral red

blue
Detect Lysine
decarboxylation
Lactose (+) = pink (+) = violet Non-
fermentation (-) = colorless ❖ E. coli = pathogenic:
metallic Orange (low
green sheen pH)
(-) = colorless Green to
orange

Pathogenic
(Salmonella &
Shigella):
Green to blue
green
Sulfur source Sodium Sodium Sodium
thiosulfate thiosulfate thiosulfate
Detect H2S Ferric Ferric Ferric
Production ammonium ammonium ammonium
citrate citrate citrate

ENTEROBACTERIACEAE
H2S (+) UREASE (+) INDOLE (+) MR (-) VP (+) PD (+) MAN (-)
Edwardsielleae Citrobacteriaceae Escherichieae Klebsiellae Klebsiellae Proteeae Edwardsielleae
Citrobacteriaceae Klebsiellae Edwardsielleae
Salmonelleae Proteeae Citrobacteriaceae
Proteeae Yersiniae Proteeae
Yersiniae