Download - Bacterial Culture Media in Plate & Tube
BACTERIOLOGY 231 PLATE MEDIA
MEDIA CARBOHYDRATE INHIBITOR pH INDICATOR RESULT
BAP Blood agar
Outdated human blood or 5% defribinated sheep blood
(preferred)
Trypticase Soy Agar or Nutrient agar
{ Nonselective agar for Hemolysis Differentiation }
PINHEAD – Staphylococcus PINPOINT - Streptococcus
Alpha hemolysis
Incomplete or partial lysis with greening S. pneumoniae
Beta hemolysis
Complete lysis with colorless zone [diffusion of hemoglobin into the surrounding medium] S. pyogenes S. agalactiae
Gamma hemolysis
Nonhemolytic
CAP Chocolate agar
Lysed red blood cells
{Nonselective agar for Hemolysis Differentiation}
PINHEAD – Staphylococcus PINPOINT – Streptococcus
**S. aureus - 1° src. of X factor
Hemolytic
Trypticase Soy Agar or Nutrient agar
X factor (hemin)
V factor or NAD (nicotinamide adenine dinucletide,
coenzyme)
Nonhemolytic
PEA Phenylethyl alcohol agar
{Selective & Differential agar} Gram (-) inhibitor
Inhibits swarming of Proteus
Bacillus anthracis (24° incubation) **Most Bacillus sp. grow
MSA Mannitol salt agar
{Selective & Differential agar} Inhibits Gram (-)
Mannitol 7.5% NaCl Phenol red
✔ Yellow colonies [pH 6.8]
Coagulase (+) S. aureus Some Micrococcus & Enterococcus
Beef extract
✖Small red colonies with red/purple zone [pH 8.4]
Coagulase (-) Staph. sp. (episodemidis/saprophiticus) Most Gram (-)
EMB Eosin methylene blue agar
{Selective & Differential agar} Inhibits Gram (+)
Lactose
Eosin Methylene blue
✔Pink to purple colonies with Green metallic sheen
E. coli (Rapid Lactose Fermenter - RLF)
✔Pink to purple colonies with Green metallic sheen w/ or w/o black center (fisheye)
Enterobacter aerogenes (RLF) Motile*
✔ Pink to purple colonies, mucoid
Klebsiella pneumonia (RLF) Nonmotile*
✖Yellow to colorless (pink) colonies (NLF)
Salmonella typhi Shigella sonnei Proteus vulgaris Psedomonas aeroginosa
MAC MacConkey agar
{Differential agar} Inhibits Gram
(+)
Lactose Crystal violet
Bile salts Neutral red
✔Pink to red colonies [pH <6.8]
RLF: Escherichia, Enterobacter, Klebsiella LLF: Citrobacter, Serratia, Hafnia alvei, Y. enterocolitica, S. arizonae, S. sonnei
✖Yellow to colorless colonies [pH 6.8-8.0]
NLF: Proteus, Providencia, Morganella, Edwardsiella Salmonella (r/o) arizonae Shigella (r/o) sonnei Yersenia (r/o) enterocolitica
HEA Hektoen Enteric agar
{Selective & Differential agar} Inhibits normal flora coliform
of the lower GI tract
[Salmonella and Shigella species from other Gram (-)
enteric organisms]
Lactose Sucrose Salicin
Bile salts Citrate
Bromthymol blue
[H2S: ferric ammonium
chitrate; sodium
thiosulfate]
✔Yellow to orange w/ black center
Citrobacter freundii Salmonella arizonae
✔Yellow to orange w/o black center
E. coli Klebsiella pneumonia Enterobacter aerogenes Shigella sonnei Serratia marcesens
✖Blue to green w/o black center
Providencia rettegri Morganella morganii Shigella dysenteriae
✖Blue to green w/ black center
Salmonella typhi Proteus vulgaris Proteus mirabilis
XLD Xylose-lysine deoxycholate
{Selective & Differential agar} Inhibits normal flora coliforms
[Partially E. coli & swarming of
Proteus]
Xylose Na+
Deoxycholate
Phenol red [H2S: ferric ammonium
chitrate; sodium
thiosulfate]
[Lysine AA → Lysine
decarboxylase]
(+)LDC = CADAVERIN
(pH)
✔ Yellow colonies w/ black center (acid)
Citrobacter freundii Proteus vulgaris Proteus mirabilis
✔Yellow colonies w/o black center (acid)
E. coli Klebsiella pneumonia Enterobacter aerogenes Providencia retteri Serratia marcesens
✖Red colonies w/ black center (alk)
Salmonella typhi
✖Red colonies w/o black center (alk)
Shigella dysenteriae
SSA Salmonella-Shigella agar
{Selective & Differential agar} Inhibition of normal flora
coliforms Inhibits Gram (+) & (-)
Lactose Bile salts Brilliant green
Neutral red [H2S: ferric ammonium
chitrate; sodium
thiosulfate]
✔Pink to red colonies (LF)
Normal flora coliforms *Shigella sonnei *Salmonella arizonae (w/ black center)
✖Yellow to colorless w/o black center
Shigella dysenteriae Shigella flexneri Shigella boydii
✖Yellow to colorless w/ black center
Salmonella typhi
TCBS Thiosulfate Citrate Bile salt Sucrose agar
{Selective & Differential agar} Isolate Vibrio sp.
Sucrose
Sodium citrate Sodium
thiosulfate Oxgall
Bromthymol blue
[H2S: ferric ammonium
chitrate; sodium
thiosulfate]
✔Yellow colonies V. cholera V. alginolyticus
✖Blue-green colonies V. parahaemolyticus V. vulnificus Aeromonas Psuedomonas
BSA Bismuth sulfate agar
{Selective & Differential agar} Isolate Salmonella sp.
Inhibit Gram (+), lactose fermenter & shigellase
Bismuth sulfate Brilliant green
(Inhibts other
enteric)
[H2S: ferric sulfide] soluble
✖ Black colonies with metallic sheen (w/ ring)
Salmonella typhi
✔Yellow to orange colonies
S. gallinarum S. choleraesuis S. paratyphi
Corynebacterium diptheriae
CYSTEINE TELLURITE AGAR (CTA) >gray to black colonies MODIFIED TYNSDALE AGAR >black colonies with brown halos LOEFFLER SERUM SLANT >enhance METACHROMATIC granules
Francisella tularensis BLOOD CYSTEIN GLUCOSE AGAR
Legionella pneumophilia BUFFERED CHARCOAL YEASE EXTRACT (BYCE) With L-cysteine
Clostridium difficile CYCLOSERINE CEFOXITIN FRUCTOSE AGAR (CCFA)
Prevotella melaninogenica (Brucella) LAKED KANAMYCIN VANCOMYCIN 5% SHEEP BLOOD AGAR (LKV)
5-6 wks: LEPTOSPIRA 3-4 wks: BRUCELLA 2-3 wks: MYCOBACTERIUM
BIPHASIC (2 phased media) >SEPTI-CHECK & OLD CASTAÑEDA *ideal for airborne microbes
BACTERIOLOGY 231
TUBE MEDIA
MEDIA FORMATION CARBOHYDRATE pH INDICATOR H2S INDICATOR RESULT
TSI Triple Sugar
Iron agar Butt Slant
Glucose [Butt] Lactose [Slant] Sucrose [Slant]
Phenol red Ferrous
ammonium sulfate
A/AG E. coli Enterobacter aerogenes Pantoea agglomerans Klebsiella pneumonia Serratia Salmonella arizonae
Decarboxylation (slant) = Alkaline (red)
Legend: A(yellow); K(red); G(gas production); +(H2S production)
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
MEDIA FORMATION AMINO ACID CARBOHYDRATE pH
INDICATOR H2S
INDICATOR RESULT
LIA Lysine Iron
Agar Butt Slant Lysine Glucose
Bromcresol purple
Ferric ammonium
citrate K/K
E. coli Enterobacter aerogenes Klebsiella pneumonia Serratia marcescens Hafnia alvei
Legend: K(purple); A(yellow); R(red); +(H2S production)
K/A
Pantoea agglomerans Citobacter freundii Shigella dysenteriae Shigella flexneri Shigella boydii
R/A *deaminase(+)
Proteus Providencia Morganella
K/AG+
Salmonella typhimurium Salmonella arizonae Salmonella typhi Salmonella parayphi Salmonella choleraesuis Edwardsiella tarda
MEDIA PRINCIPLE FORMATION MEDIUM REAGENT POSITIVE NEGATIVE
Indol Production
Test
The organism oxidized tryptophan
by trytophanase ↓
tryptophan ↓
pyruvic acid ↓
amino acid ↓
indol
Butt Tryptone
broth
①Ehrlich’s Add: EtOH and/or
XYLENE (xtract indol) then p-dab
②Kovac’s p-aminodimethyl
benzaldehyde (p-dab)
③Spot Indole: rapid detection –
filter paper:1% pdab in 4 hours.
Red Ring Yellow/Brown
Ring
Methyl Red Test
MIXED ACID
PATHWAY
The organism ferments glucose to
pyruvic acid resulting in mixed acid
products leading to decreased pH (4.5-5.5) of the medium and takes the color
of the indicator
Butt
MRVP (Methyl
Red Voges Proskauer)
medium
Methyl red indicator
Red solution
Yellow solution
Voges Proskauer
Test
BUTYLENE GLYCOL
PATHWAY
The organism ferments glucose
producing acetylmethylcarbinol
(acetoin) and converted to
dimethyl/diacetyl in the presence of
oxygen and KOH.
KOH → reacts w/ GUANIDINE = color
Butt MRVP
VP Reagent A: Alpha napthol in absolute methyl
alcohol
VP Reagent B: 40% KOH with 0.3%
creatine
Pink to Red color
Yellow or copper-like
MEDIA PRINCIPLE FORMATION MEDIUM INDICATOR POSITIVE NEGATIVE
Citrate Utilization
Test
The organism utilizes the citrate (sole src. of carbon) producing
ammonia and converted to ammonium
hydroxide. This alkaline compound raises the pH of the medium and takes
the blue color.
Slant
SCA (Simmon
Citrate Agar)
Bromthymol blue
Prussian blue color
Green color
Urease Test
Urease splits the urea molecules into
ammonia, carbon dioxide and water. Ammonia reacts in
solution to form alkaline compound,
ammonium carbonate, which
results in an increased pH of the medium and a color
change in the indicator to pink-red.
Butt (Broth) Urea broth
Phenol red
Pink-red No color change
Sulfide indole
motility (SIM)
Butt (Semi-
solid)
H2S: ferric ammonium chitrate; sodium thiosulfate
Motile (growth
pattern – away from inoculation
line)
Nonmotile (growth
pattern is confined)
H2S production (Blackening of the butt)
Indol production (red ring)
Brilliant Green Bile
Lactose Broth
Water Analysis Testing >sanitary supply of water
>suitability for general use
Method: Multiple Fermentation Tube Technique
PRESUMPTIVE TEST: LACTOSE BROTH CONFIRMED TEST: BGBL, EMB, ENDO AGAR
COMPLETED TEST: LACTOSE BROTH
Uninocculated
Positive E. coli + Gas Production
+ Turbidity