231 CARBOHYDRATE Alpha hemolysis INHIBITOR pH INDICATOR

P RESULT Incomplete or partial lysis with greening

M D

MEDIA

BAP Blood agar

Outdated human blood or 5% defribinated sheep blood (preferred) {Nonselective Differential agar}

Beta hemolysis

Complete lysis with colorless zone [diffusion of hemoglobin into the surrounding medium]

Trypticase Soy Agar or Nutrient agar Gamma hemolysis Nonhemolytic

Hemolytic

CAP Chocolate agar

Lysed red blood cells

Trypticase Soy Agar or Nutrient agar {Nonselective Differential agar} Nonhemolytic

X factor (hemin) V factor or NAD ( nicotinamide adenine dinucletide, coenzyme) {Selective agar} Staphylococcus sp.

PEA Phenylethyl

{Selective & Differential agar} Gram (-) inhibitor

alcohol agar

Inhibits swarming of Proteus

MSA Mannitol salt agar Mannitol Small red colonies with red/purple zone [pH 8.4] Green metallic sheen (LF) 7.5% NaCl Phenol red

{Selective & Differential agar} Inhibits Gram (+)

Yellow colonies [pH 6.8]

Coagulase (+) Staph. sp. Some Micrococcus & Enterococcus Most Gram (-) Coagulase (-) Staph. Sp.

Beef extract

EMB Eosin methylene blue agar

E. coli (lactose fermenter)

{Selective & Differential agar} Inhibits Gram (+) Lactose Eosin Methylene blue

Enterobacter aerogenes (RLF) Klebsiella pneumoniae Salmonella typhi Shigella sonnei Proteus vulgaris Psedomonas aeroginosa Lactose fermenting Gram (-) Pink to red colonies [pH <6.8]

Green metallic sheen w/ fisheye Large purple mucoid (LF) Yellow to colorless (pink) colonies (NLF)

MAC MacConkey agar Lactose

{Differential agar}

Crystal violet Bile salts

Neutral red

Yellow to colorless colonies [pH 6.8-8.0]

Non-lactose fermenting Gram (-)

HEA Hektoen enteric agar Blue to green w/o black center Lactose Sucrose Salicin Bile salts Citrate Yellow to orange w/ black center Yellow to orange w/o black center Bromthymol blue

Blue to green w/ black center

{Selective & Differential agar} Inhibits normal flora coliform of the lower GI tract

[Salmonella and Shigella species from other Gram (-) enteric organisms]

XLD Xylose-lysine deoxycholate Yellow colonies w/o black center

Yellow colonies w/ black center

{Selective & Differential agar} Inhibits normal flora coliforms Xylose Deoxycholate

[Partially E. coli & swarming of Proteus]

Phenol red [H2S: ferric ammonium chitrate; sodium thiosulfate]

Providencia rettegri Morganella morganii Shigella dysenteriae Salmonella typhi Proteus vulgaris Proteus mirabilis Citrobacter freundii Salmonella arizonae E. coli Klebsiella pneumonia Enterobacter aerogenes Shigella sonnei Serratia marcesens Citrobacter freundii Proteus vulgaris Proteus mirabilis E. coli Klebsiella pneumonia Enterobacter aerogenes Providencia retteri Serratia marcesens Salmonella typhi Shigella dysenteriae Normal flora coliforms

SSA SalmonellaShigella agar Lactose Bile salts Brilliant green

{Selective & Differential agar} Inhibition of normal flora coliforms Inhibits Gram (+) & (-)

Red colonies w/ black center Red colonies w/o black center Pink to red colonies w/o black center Yellow to colorless w/o black center Neutral red [H2S: ferric ammonium chitrate; sodium thiosulfate] Yellow to colorless w/ black center Yellow colonies Blue-green colonies

Shigella dysenteriae Shigella flexneri Shigella boydii Salmonella typhi

TCBS Thiosulfate Citrate Bile salt Sucrose agar Sucrose

{Selective & Differential agar} Isolate Vibrio sp.

Sodium citrate Sodium thiosulfate Oxgall

Bromthymol blue [H2S: ferric ammonium chitrate; sodium thiosulfate]

V. cholera V. alginolyticus V. parahaemolyticus V. vulnificus Aeromonas Psuedomonas

BSA Bismuth sulfate agar Bismuth sulfate Brilliant green [H2S: ferric sulfide] soluble No growth S. gallinarum S. choleraesuis S. paratyphi Other enterics

{Selective & Differential agar} Isolate Salmonella sp.

Black colonies with metallic cheen Light green colonies

Salmonella typhi

Inhibit Gram (+), lactose fermenter & shigellase

231 CARBOHYDRATE Glucose Lactose Sucrose A/AG+ or K/AG+ (5) A/AG or K/AG (4a) pH INDICATOR H2S INDICATOR Phenol red Ferrous sulfate RESULT A/AG (4)

U M D MEDIA TSI Triple Sugar Iron agar

FORMATION Butt Slant

K/AG+

A/A or K/A K/A (2)

K/A+ (3) K/K (1)

E. coli Enterobacter aerogenes Pantoea agglomerans Klebsiella pneumonia Citobacter freundii Salmonella arizonae Citrobacter koseri Serratia marcescens Hafnia alvei Proteus vulgaris Proteus mirabilis Salmonella paratyphi Salmonella choleraesuis Edwardsiella tarda Yersinia entericolitica Shigella sonnei Shigella dysenteriae Shigella flexneri Shigella boydii Salmonella typhi Pseudonmonas aeroginosa Alcaligenes

MEDIA

FORMATION

AMINO ACID

CARBOHYDRATE

pH INDICATOR

H2S INDICATOR

RESULT

LIA Lysine Iron Agar [Lysine deamination] K/A

Butt Slant

Lysine

Glucose

Bromcresol purple

Ferric ammonium citrate

K/K

R/A

K/AG+

E. coli Enterobacter aerogenes Klebsiella pneumonia Serratia marcescens Hafnia alvei Pantoea agglomerans Citobacter freundii Shigella dysenteriae Shigella flexneri Shigella boydii Proteus Providencia Morganella 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 to indol by trytophanase.

Butt

Tryptone broth

Ehrlichs or Kovacs (paminobenzaldehyde)

Red Ring

Yellow/Brown Ring

Methyl Red Test

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 VP Reagent A: Alpha napthol in absolute methyl alcohol VP Reagent B: KOH with creatine

Butt

MRVP (Methyl Red Voges Proskauer) medium

Methyl red indicator

Red solution

Yellow solution

Voges Proskauer Test

The organism ferments glucose producing acetylmethylcarbinol (acetoin) and converted to diacetyl in the presence of oxygen and KOH.

Pink to Red color

Yellow or copper-like

Citrate Utilization Test

The organism utilizes the citrate 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 (indicator)

Prussian blue color

Green color

Urease Test Urea broth Phenol red (indicator)

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 pinkred.

Butt (Broth)

Pink-red

No color change

Sulfide indole motility (SIM)

Butt (Semisolid)

Motile Nonmotile (growth (growth pattern pattern is away from confined) inoculation line) H2S production (Blackening of the butt) Indol production (red ring)