BACTERIOLOGY 231

PLATE MEDIA
MEDIA CARBOHYDRATE INHIBITOR pH INDICATOR RESULT
BAP Alpha hemolysis Incomplete or partial lysis with
Blood agar greening

S. pneumoniae

Beta hemolysis Complete lysis with colorless

Outdated human blood or 5% zone [diffusion of hemoglobin
defribinated sheep blood into the surrounding medium]
{ Nonselective agar for Hemolysis Differentiation }
(preferred)
PINHEAD – Staphylococcus S. pyogenes
Trypticase Soy Agar or S. agalactiae
PINPOINT - Streptococcus
Nutrient agar

Gamma hemolysis Nonhemolytic

CAP Hemolytic
Chocolate Lysed red blood cells
agar
{Nonselective agar for Hemolysis Differentiation}
Trypticase Soy Agar or
Nutrient agar
PINHEAD – Staphylococcus
PINPOINT – Streptococcus Nonhemolytic
X factor (hemin)
V factor or NAD (nicotinamide **S. aureus - 1° src. of X factor
adenine dinucletide,
coenzyme)
PEA Bacillus anthracis
Phenylethyl (24° incubation)
alcohol agar {Selective & Differential agar} **Most Bacillus sp. grow
Gram (-) inhibitor

Inhibits swarming of Proteus

MSA ✔ Yellow colonies Coagulase (+) S. aureus

Mannitol salt [pH 6.8]
agar Some Micrococcus &
Enterococcus
{Selective & Differential agar}
Inhibits Gram (-)

Mannitol 7.5% NaCl Phenol red

✖Small red colonies Coagulase (-) Staph. sp.
with red/purple zone (episodemidis/saprophiticus)
[pH 8.4] Most Gram (-)

Beef extract

EMB ✔Pink to purple E. coli (Rapid Lactose

Eosin colonies with Green Fermenter - RLF)
methylene metallic sheen
blue agar
{Selective & Differential agar}
Eosin
Inhibits Gram (+) Lactose
Methylene blue
 ✔Pink to purple Enterobacter aerogenes (RLF)
colonies with Green
metallic sheen w/ or Motile*
w/o black center
(fisheye)
✔ Pink to purple Klebsiella pneumonia (RLF)
colonies, mucoid
Nonmotile*

✖Yellow to colorless Salmonella typhi

(pink) colonies (NLF) Shigella sonnei
Proteus vulgaris
Psedomonas aeroginosa
MAC ✔Pink to red colonies RLF: Escherichia,
MacConkey [pH <6.8] Enterobacter, Klebsiella
{Differential agar}
agar LLF: Citrobacter, Serratia,
Inhibits Gram
Hafnia alvei, Y. enterocolitica,
S. arizonae, S. sonnei
Crystal violet
Lactose Neutral red ✖Yellow to colorless NLF: Proteus, Providencia,
Bile salts
colonies [pH 6.8-8.0] Morganella, Edwardsiella

Salmonella (r/o) arizonae

(+)
Shigella (r/o) sonnei
Yersenia (r/o) enterocolitica
HEA ✔Yellow to orange w/ Citrobacter freundii
Hektoen black center Salmonella arizonae
Enteric agar ✔Yellow to orange w/o E. coli
{Selective & Differential agar} Bromthymol black center Klebsiella pneumonia
Inhibits normal flora coliform blue Enterobacter aerogenes
of the lower GI tract Lactose [H2S: ferric Shigella sonnei
Bile salts
Sucrose ammonium Serratia marcesens
Citrate
[Salmonella and Shigella Salicin chitrate; ✖Blue to green w/o Providencia rettegri
species from other Gram (-) sodium black center Morganella morganii
enteric organisms] thiosulfate] Shigella dysenteriae
✖Blue to green w/ Salmonella typhi
black center Proteus vulgaris
Proteus mirabilis
XLD Phenol red ✔ Yellow colonies w/ Citrobacter freundii
Xylose-lysine [H2S: ferric black center (acid) Proteus vulgaris
deoxycholate ammonium Proteus mirabilis
chitrate; ✔Yellow colonies w/o E. coli
sodium black center (acid) Klebsiella pneumonia
{Selective & Differential agar} Enterobacter aerogenes
thiosulfate]
Inhibits normal flora coliforms Providencia retteri
Na+
Xylose Serratia marcesens
Deoxycholate [Lysine AA →
[Partially E. coli & swarming of
Lysine
Proteus]
decarboxylase
] ✖Red colonies w/ black Salmonella typhi
(+)LDC = center (alk)
CADAVERIN ✖Red colonies w/o Shigella dysenteriae
(pH) black center (alk)
SSA ✔Pink to red colonies Normal flora coliforms
Salmonella- (LF)
Shigella agar Neutral red *Shigella sonnei
{Selective & Differential agar}
[H2S: ferric *Salmonella arizonae (w/
Inhibition of normal flora Bile salts
ammonium black center)
coliforms Lactose Brilliant
chitrate; ✖Yellow to colorless Shigella dysenteriae
Inhibits Gram (+) & (-) green
sodium w/o black center Shigella flexneri
thiosulfate] Shigella boydii
✖Yellow to colorless Salmonella typhi
w/ black center
TCBS ✔Yellow colonies V. cholera
Thiosulfate V. alginolyticus
Citrate Bile ✖Blue-green colonies V. parahaemolyticus
salt Sucrose V. vulnificus
agar Bromthymol
Sodium blue Aeromonas
{Selective & Differential agar} citrate [H2S: ferric Psuedomonas
Isolate Vibrio sp. Sucrose Sodium ammonium
thiosulfate chitrate;
Oxgall sodium
thiosulfate]
BSA ✖ Black colonies with Salmonella typhi
Bismuth metallic sheen
sulfate agar (w/ ring)

Bismuth
{Selective & Differential agar} sulfate
Isolate Salmonella sp. Brilliant [H2S: ferric
green sulfide]
Inhibit Gram (+), lactose soluble
✔Yellow to orange S. gallinarum
fermenter & shigellase (Inhibts other S. choleraesuis
colonies
enteric) S. paratyphi

CYSTEINE TELLURITE AGAR (CTA)

>gray to black colonies

MODIFIED TYNSDALE AGAR

Corynebacterium diptheriae
>black colonies with brown halos

LOEFFLER SERUM SLANT

>enhance METACHROMATIC granules
Francisella tularensis BLOOD CYSTEIN GLUCOSE AGAR
BUFFERED CHARCOAL YEASE EXTRACT
Legionella pneumophilia (BYCE)
With L-cysteine
CYCLOSERINE CEFOXITIN FRUCTOSE AGAR
Clostridium difficile
(CCFA)
(Brucella) LAKED KANAMYCIN
Prevotella melaninogenica VANCOMYCIN 5% SHEEP BLOOD AGAR
(LKV)
5-6 wks: LEPTOSPIRA BIPHASIC (2 phased media)
3-4 wks: BRUCELLA >SEPTI-CHECK & OLD CASTAÑEDA
2-3 wks: MYCOBACTERIUM *ideal for airborne microbes
BACTERIOLOGY 231
TUBE MEDIA
MEDIA FORMATION CARBOHYDRATE pH INDICATOR H2S INDICATOR RESULT
A/AG E. coli
Enterobacter aerogenes
TSI Glucose [Butt] Ferrous Pantoea agglomerans
Triple Sugar Butt Slant Lactose [Slant] Phenol red ammonium Klebsiella pneumonia
Iron agar Sucrose [Slant] sulfate
Serratia
Salmonella arizonae
Decarboxylation (slant) = Alkaline (red) A/AG+ or K/AG+ Citobacter freundii
Salmonella arizonae

Hafnia alvei
Morganella morganii
Morganella freundii
A/AG or K/AG Citrobacter koseri
Serratia marcescens
Hafnia alvei
K/AG+ Proteus vulgaris
Proteus mirabilis
Salmonella paratyphi
Salmonella choleraesuis
Edwardsiella tarda
A/A or K/A Yersinia entericolitica
Shigella sonnei
K/A Shigella dysenteriae
Shigella flexneri
Shigella boydii
K/A+ Salmonella typhi
K/K Pseudonmonas aeroginosa
Alcaligenes

Legend: A(yellow); K(red); G(gas production); +(H2S production)

 pH H2S
MEDIA FORMATION AMINO ACID CARBOHYDRATE RESULT
INDICATOR INDICATOR
E. coli
LIA Ferric Enterobacter aerogenes
Bromcresol
Lysine Iron Butt Slant Lysine Glucose ammonium K/K Klebsiella pneumonia
purple
Agar citrate Serratia marcescens
Hafnia alvei
Pantoea agglomerans
Citobacter freundii
K/A Shigella dysenteriae
Shigella flexneri
Shigella boydii
Proteus
R/A
Providencia
*deaminase(+)
Morganella
Salmonella typhimurium
Salmonella arizonae
Salmonella typhi
K/AG+ Salmonella parayphi
Salmonella choleraesuis
Edwardsiella tarda
Legend: K(purple); A(yellow); R(red); +(H2S production)
 MEDIA PRINCIPLE FORMATION MEDIUM REAGENT POSITIVE NEGATIVE
The organism ①Ehrlich’s
oxidized tryptophan Add: EtOH and/or
by trytophanase XYLENE (xtract
↓ indol) then p-dab
tryptophan ②Kovac’s
Indol
↓ Tryptone p-aminodimethyl Yellow/Brown
Production Butt Red Ring
pyruvic acid broth benzaldehyde Ring
Test
↓ (p-dab)
amino acid ③Spot Indole:
↓ rapid detection –
indol filter paper:1% pdab
in 4 hours.

The organism
ferments glucose to
Methyl
pyruvic acid resulting MRVP
Red Test
in mixed acid (Methyl
Red Yellow
products leading to Butt Red Voges Methyl red indicator
MIXED solution solution
decreased pH (4.5- Proskauer)
ACID
5.5) of the medium medium
PATHWAY
and takes the color
of the indicator

The organism
ferments glucose
producing VP Reagent A:
Voges
acetylmethylcarbinol Alpha napthol in
Proskauer
(acetoin) and absolute methyl
Test
converted to alcohol Pink to Yellow or
Butt MRVP
dimethyl/diacetyl in Red color copper-like
BUTYLENE
the presence of VP Reagent B:
GLYCOL
oxygen and KOH. 40% KOH with 0.3%
PATHWAY
creatine
KOH → reacts w/
GUANIDINE = color
 MEDIA PRINCIPLE FORMATION MEDIUM INDICATOR POSITIVE NEGATIVE

The organism utilizes

the citrate (sole src.
of carbon) producing
ammonia and
SCA
Citrate converted to
(Simmon Prussian
Utilization ammonium Slant Bromthymol blue Green color
Citrate blue color
Test hydroxide. This
Agar)
alkaline compound
raises the pH of the
medium and takes
the blue color.

Urease splits the

urea molecules into
ammonia, carbon
dioxide and water.
Ammonia reacts in
solution to form
Urease alkaline compound, Urea No color
Butt (Broth) Phenol red Pink-red
Test ammonium broth change
carbonate, which
results in an
increased pH of the
medium and a color
change in the
indicator to pink-red.
Motile
(growth Nonmotile
pattern – (growth
away from pattern is
Sulfide inoculation confined)
H2S: ferric
indole Butt (Semi- line)
ammonium chitrate;
motility solid)
sodium thiosulfate H2S production (Blackening
(SIM)
of the butt)

Indol production (red ring)

 Water Analysis Testing
>sanitary supply of water
>suitability for general use

Method: Multiple Fermentation Tube Technique

Uninocculated
PRESUMPTIVE TEST: LACTOSE BROTH
CONFIRMED TEST: BGBL, EMB, ENDO AGAR
Brilliant COMPLETED TEST: LACTOSE BROTH
Green Bile
Lactose
Broth

Positive E. coli
+ Gas Production
+ Turbidity