Schleifer-Krไmer Agar

(SK Agar)

Composition per liter:

Agar .............................................................. ...13.0g

Glycerol ........................................................ ...10.0g

Sodium pyruvate............................................. ..10.0g

Pancreatic digest of casein............................... ..10.0g

Beef extract...................................................... ...5.0g

Yeast extract...................................................... ..3.0g

Potassium isothiocyanate................................. .2.25g

LiCl.................................................................. ...2.0g

Na2HPO4*2H2O............................................... ...0.9g

NaH2PO4*H2O.................................................. ..0.6g

Glycine............................................................... .0.5g

NaN3 solution ............................................ .. 10.0mL

pH 7.2 ± 0.2 at 25oC

NaN3 Solution:

Composition per 10.0mL:

NaN3 ........................................................... ...0.045g

Preparation of NaN3 Solution: Add NaN3 to distilled/deionized  water  and  bring  volume  to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept NaN3 solution, to distilled/deionized water and
bring volume to 990.0mL. Mix thoroughly. Adjust
pH to 7.2. Gently heat and bring to boiling. Auto-
clave for 15 min at 15 psi pressure-121oC. Cool to
45o-50oC. Aseptically add sterile NaN3 solution.
Mix thoroughly. Pour into sterile Petri dishes or dis-
tribute into sterile tubes.

Use: For the isolation and cultivation of Staphylo-
coccus species.


Selenite Broth

(Selenite Broth, Lactose)

(Selenite F Enrichment Medium)

(Sodium Biselenite Medium)
   (Sodium Hydrogen Selenite Medium)
Composition per liter:

Na2HPO4........................................................ ..10.0g

Pancreatic digest of casein.................................. .5.0g

Lactose............................................................... .4.0g

NaHSeO3*5H2O .............................................. ..4.0g

pH 7.0 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems and a prepared medium from Oxoid Unipath.

Caution: Sodium biselenite is toxic and a potential

teratogen and care must be taken to avoid inhalation of the powdered dye, contact with the skin, or inges-
tion, especially in pregnant laboratory workers.

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling. Do
not autoclave. Distribute into sterile tubes in 10.0mL
volumes.

Use: For the isolation and enrichment of Salmonella species from clinical specimens.


Selenite Broth Base, Mannitol

Composition per liter:

Na2HPO4........................................................ ..10.0g

Peptone.............................................................. .5.0g

Mannitol........................................................... ..4.0g

NaHSeO3*5H2O .............................................. ..4.0g

pH 7.1 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from Oxoid Unipath.

Caution: Sodium selenite is toxic and a potential
teratogen and care must be taken to avoid inhalation
of the powdered dye, contact with the skin, or inges-
tion, especially in pregnant laboratory workers.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat. Do not autoclave. Dis-
tribute into sterile tubes in 10.0mL volumes. Sterilize for 10 min at 0 psi pressure-100oC.

Use: For the isolation and cultivation of Salmonella typhi and Salmonella paratyphi.


Selenite Cystine Broth

Composition per liter:

Na2HPO4........................................................ ..10.0g

Pancreatic digest of casein................................. .5.0g

Lactose............................................................... .4.0g

Na2SeO3*5H2O.................................................. .4.0g

L-Cystine ........................................................ ..0.02g

pH 7.0 ± 0.2 at 25oC

Source: This medium is available as a premixed
powder from  BD Diagnostic Systems and Oxoid Un-
ipath.

Caution: Sodium selenite is toxic and a potential
teratogen and care must be taken to avoid inhalation
of the powdered dye, contact with the skin, or inges-
tion, especially in pregnant laboratory workers.

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat. Do not autoclave. Dis- 

tribute into sterile tubes in 10.0mL volumes. Sterilize

for 15 min at 0 psi pressure-100oC.

Use: For the isolation and cultivation of Salmonella species from feces and other specimens.


Selenite F Broth

Composition per liter:

KH2PO4........................................................... ...7.0g

Pancreatic digest of casein.................................. .5.0g

Lactose............................................................... .4.0g

Na2SeO3*5H2O.................................................. .4.0g

Na2HPO4........................................................... .3.0g

pH 7.0 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Caution: Sodium selenite is toxic and a potential
teratogen and care must be taken to avoid inhalation
of the powdered dye, contact with the skin, or inges-
tion, especially in pregnant laboratory workers.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat. Do not autoclave. Dis-
tribute into sterile tubes in 10.0mL volumes. Sterilize for 30 min at 0 psi pressure-100oC.

Use: For the isolation and cultivation of Salmonella species from feces and other specimens.


Semisolid Brucella Broth

Composition per liter:

Peptone ........................................................... .10.0g

Pancreatic digest of casein............................... ..10.0g

NaCl.................................................................. ..5.0g

Yeast extract...................................................... ..2.0g

Agar ................................................................. ..1.6g

D-Glucose........................................................ ...1.0g

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Auto-
clave for 15 min at 15 psi pressure-121oC.

Use: For the cultivation of Campylobacter species.


Semisolid Medium for Motility

Composition per liter:

Biosate .............................................................. .5.0g

Polypeptone ..................................................... ..5.0g

NaCl.................................................................. ..5.0g

Agar ................................................................. ..4.0g

Myosate........................................................... ...1.5g

Triphenyltetrazolium chloride solution .......... . 2.5mL

pH 6.9-7.1at 25oC

Triphenyltetrazolium Chloride Solution:

Composition per 10.0mL:

Triphenyltetrazolium chloride ........................... .0.1g

Ethanol (95% solution).............................. ...10.0mL

Preparation of Triphenyltetrazolium Chlo-
ride Solution: Add triphenyltetrazolium chloride to 10.0mL of ethanol. Mix thoroughly.

Preparation of Medium: Add components, ex-
cept triphenyltetrazolium chloride solution, to dis-
tilled/deionized water and bring volume to 997.5mL.
Mix thoroughly. Gently heat and bring to boiling.
Add 2.5mL of triphenyltetrazolium chloride solu-
tion. Mix thoroughly. Distribute into tubes in 10.0mL
volumes. Autoclave for 15 min at 15 psi pressure-
121oC.

Use: For the differentiation of bacteria based on mo-
tility.


Sensitest Agar

Composition per liter:

Pancreatic digest of casein.............................. ..11.0g

Agar ................................................................. ..8.0g

Buffer salts........................................................ .3.3g

Peptone.............................................................. .3.0g

NaCl................................................................. ..3.0g

Glucose ........................................................... ...2.0g

Starch .............................................................. ...1.0g

Nucleoside bases............................................ ..0.02g

Thiamine...................................................... ..0.02mg

pH 7.4 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from Oxoid Unipath.

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es.

Use: For the performance of antibiotic sensitivity as-
says.


Serratia Differential Medium

(SD Medium)

Composition per 102.0mL:

Solution A.................................................. ..92.0mL

Solution B .................................................. ..10.0mL

pH 6.7 ± 0.2 at 25oC

Solution A:

Composition per 92.0mL:

Yeast extract..................................................... ..1.0g

L-Ornithine ........................................................ .1.0g

NaCl................................................................. ..0.5g


 

Agar ................................................................. ..0.4g

Irgasan inhibitor............................................. . 1.0mL

Indicator solution.......................................... .. 1.0mL

Preparation of Solution A: Add components to
distilled/deionized  water  and  bring  volume  to
92.0mL. Mix thoroughly. Adjust pH to 6.7 with 1N
NaOH.

Irgasan Inhibitor:

Composition per 100.0mL:

Irgasan-DP-300 (4,2ด, 4ด-trichloro-

2-hydroxydiphenylether)........................... ...0.1g

NaOH (1N solution)................................... .. 10.0mL

Preparation of Irgasan Inhibitor: Add irgasan to 10.0mL of NaOH solution. Mix thoroughly. Gen-
tly heat to dissolve. Bring volume to 100.0mL with distilled/deionized water.

Indicator Solution:

Composition per 100.0mL:

Bromthymol Blue ............................................. ..0.2g

Phenol Red......................................................... .0.1g

Preparation of Indicator Solution: Add com-
ponents to 50.0mL of distilled/deionized water. Mix thoroughly for 1h. Bring volume to 100.0mL with distilled/deionized water.

Solution B:

Composition per 10.0mL:

L-Arabinose...................................................... ...1.0g

Preparation of Solution B: Add L-arabinose to distilled/deionized  water  and  bring  volume  to 10.0mL. Mix thoroughly.

Preparation of Medium: Combine 92.0mL of
solution A with 10.0mL of solution B. Mix thorough-
ly. Distribute into tubes. Autoclave for 15 min at 15
psi pressure-121oC. Allow tubes to cool in an up-
right position.

Use: For the cultivation and differentiation of Serratia
species based on the fermentation of arabinose and pro-
duction of ornithine decarboxylase. Serratia marce-
scens changes the medium to purple throughout the
tube. Serratia liquefaciens changes the medium to a
band of purple at the top of the tube with a green/yellow
butt. Serratia rubidaea changes the medium to yellow
throughout the tube.


Serratia Hd-MHr

Composition per liter:

Agar .............................................................. ...15.0g

K2HPO4........................................................... ...7.0g

Glucose ........................................................... ...5.0g

KH2PO4........................................................... ...3.0g

2-Methyl-DL-histidine*2HCl ............................. .1.0g

(NH4)2SO4........................................................ ..1.0g

MgSO4*7H2O.................................................. ...0.5g

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.

Use: For the cultivation and maintenance of Serratia marcescens.


Serratia Medium

(ATCC Medium 181)

Composition per liter:

Agar .............................................................. ...20.0g

Pancreatic digest of casein................................. .5.0g

Yeast extract..................................................... ..5.0g

Glucose ........................................................... ...1.0g

K2HPO4........................................................... ...1.0g

pH 7.0 ± 0.2 at 25oC

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.

Use: For the cultivation and maintenance of Serratia marcescens.


Serratia Medium

(ATCC Medium 1399)

Composition per liter:

Agar .............................................................. ...15.0g

K2HPO4........................................................... ...7.0g

Glucose ........................................................... ...5.0g

KH2PO4........................................................... ...3.0g

Casein hydrolysate............................................. .1.0g

(NH4)2SO4........................................................ ..1.0g

Yeast extract..................................................... ..1.0g

MgSO4*7H2O.................................................. ...0.1g

pH 7.0 ± 0.2 at 25oC

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.

Use: For the cultivation and maintenance of Serratia marcescens.


 

Serum Glucose Agar

(Serum Dextrose Agar)

(ATCC Medium 287)

Composition per 1060.0mL:

Agar .............................................................. ...15.0g

Peptone ........................................................... .10.0g

Beef extract...................................................... ...5.0g

NaCl.................................................................. ..5.0g

Serum-glucose solution .............................. .. 60.0mL

pH 7.3 ± 0.2 at 25oC

Serum-Glucose Solution:

Composition per 60.0mL:

D-Glucose......................................................... .10.0g

Serum (inactivated at 56oC, 30 min) ......... ... 50.0mL

Preparation of Serum-Glucose Solution: Add
glucose to 50.0mL of heat-inactivated serum. Horse
serum or ox serum may be used. Mix thoroughly. Fil-
ter sterilize.

Preparation of Medium: Add components, ex-
cept serum-glucose solution, to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly.
Gently heat and bring to boiling. Autoclave for 15
min at 10 psi pressure-115oC. Cool to 50oC. Asepti-
cally add 60.0mL of sterile serum-glucose solution.
Mix thoroughly. Pour into sterile Petri dishes or dis-
tribute into sterile tubes. Allow tubes to cool in a
slanted position.

Use: For the cultivation and maintenance of Brucel-
la species.


Serum Glucose Agar, Farrell Modified

Composition per 1086.9mL:

Agar .............................................................. ...15.0g

Peptone ........................................................... .10.0g

Beef extract...................................................... ...5.0g

NaCl.................................................................. ..5.0g

Serum-glucose solution .............................. .. 60.0mL

Bacitracin solution........................................ . 12.5mL

Cycloheximide solution............................... .. 10.0mL

Nystatin solution.......................................... ... 2.0mL

Polymyxin B solution ................................... . 1.0mL

Nalidixic acid solution.................................. ... 1.0mL

Vancomycin solution ................................... .. 0.4mL

pH 7.3 ± 0.2 at 25oC

Serum-Glucose Solution:

Composition per 60.0mL:

D-Glucose......................................................... .10.0g

Serum (inactivated at 56oC, 30 min) ......... ... 50.0mL

Preparation of Serum-Glucose Solution: Add
glucose to 50.0mL of heat-inactivated serum. Horse
serum or ox serum may be used. Mix thoroughly. Fil-
ter sterilize.


 

Bacitracin Solution:

Composition per 12.5mL:

Bacitracin................................................... ..25,000U

Preparation of Bacitracin Solution: Add baci-
tracin to distilled/deionized water and bring volume to 12.5mL. Mix thoroughly. Filter sterilize.

Cycloheximide Solution:

Composition per 100.0mL:

Cycloheximide................................................... .1.0g

Acetone......................................................... ..5.0mL

Preparation of Cycloheximide Solution: Add cycloheximide to 5.0mL of acetone. Mix thoroughly. Bring volume to 100.0mL with distilled/deionized water. Mix thoroughly. Filter sterilize.

Nystatin Solution:

Composition per 5.0mL:

Nystatin.................................................. ...250,000U

Preparation of Nystatin Solution: Add nystatin to distilled/deionized water and bring volume to 5.0mL. Mix thoroughly. Filter sterilize.

Polymyxin B Solution:

Composition per 2.0mL:

Polymyxin B ............................................ ...10,000U

Preparation of Polymyxin B Solution: Add
polymyxin B to distilled/deionized water and bring
volume to 2.0mL. Mix thoroughly. Filter sterilize.

Nalidixic Acid Solution:

Composition per 2.0mL:

Nalidixic acid................................................... ...0.1g

NaOH (0.5N solution)................................... .2.0mL

Preparation of Nalidixic Acid Solution: Add nalidixic acid to 2.0mL of NaOH solution. Mix thor-
oughly. Immediately before use, add 1.0mL of this stock solution to 9.0mL of distilled/deionized water. Mix thoroughly. Filter sterilize.

Vancomycin Solution:

Composition per 1.0mL:

Vancomycin .................................................. ...0.05g

Preparation of Vancomycin Solution: Add
vancomycin to distilled/deionized water and bring
volume to 1.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components—ex-
cept serum-glucose solution, bacitracin solution, cy-
cloheximide solution, nystatin solution, polymyxin B
solution, nalidixic acid solution, and vancomycin so-
lution—to distilled/deionized water and bring vol-
ume to 1.0L. Mix thoroughly. Gently heat and bring
to boiling. Autoclave for 15 min at 10 psi pressure-
115oC. Cool to 50oC. Aseptically add 60.0mL of
sterile serum-glucose solution, 12.5mL of sterile ba-

citracin solution, 10.0mL of sterile cycloheximide
solution, 2.0mL of sterile nystatin solution, 1.0mL of
sterile polymyxin B solution, 1.0mL of sterile nalid-
ixic acid solution, and 0.4mL of sterile vancomycin
solution. Mix thoroughly. Pour into sterile Petri dish-
es or distribute into sterile tubes. Allow tubes to cool
in a slanted position.

Use: For the selective isolation and cultivation of Brucella species.


Serum Potato Infusion Agar

Composition per 1120.0mL:

Agar .............................................................. ...15.0g

Peptone ........................................................... .10.0g

Meat extract ...................................................... ..5.0g

NaCl.................................................................. ..5.0g

Potato infusion................................................ ... 1.0L

Horse serum, heat inactivated................... ... 100.0mL

Glycerol ..................................................... .. 20.0mL

pH 6.8 ± 0.2 at 25oC

Potato Infusion:

Composition per 10.0mL:

Potatoes......................................................... ..250.0g

Preparation of Potato Infusion: Add peeled, thinly sliced potatoes to 1.0L of distilled/deionized water. Infuse overnight at 60oC. Filter through What-
man no. 1 filter paper. Bring volume to 1.0L with dis-
tilled/deionized water.

Preparation of Medium: Combine components, except horse serum. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pres-
sure-121oC. Cool to 45o-50oC. Aseptically add

100.0mL of sterile horse serum. Mix thoroughly.
Pour into sterile Petri dishes or distribute into sterile
tubes.

Use: For the cultivation of Brucella species.


Serum Tellurite Agar

Composition per liter:

Agar .............................................................. ...20.0g

Pancreatic digest of casein............................... ..10.0g

Peptic digest of animal tissue ........................ ...10.0g

NaCl.................................................................. ..5.0g

Glucose ........................................................... ...2.0g

Lamb serum ............................................... .. 50.0mL

Chapman tellurite solution............................ . 10.0mL

pH 7.5 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Chapman Tellurite Solution:

Composition per 100.0mL:

K2TeO3............................................................... .1.0g

Preparation of Chapman Tellurite Solution:

Add K2TeO3 to distilled/deionized water and bring
volume to 100.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept lamb serum and Chapman tellurite solution, to distilled/deionized  water  and  bring  volume  to 940.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 45o-50oC. Aseptically add sterile lamb serum and 10.0mL of sterile Chapman tellurite solution. Mix thoroughly. Pour into sterile Petri dish-
es or distribute into sterile tubes.

Use: For the isolation and cultivation of Corynebac-
terium species, especially in the laboratory diagnosis of diphtheria.


Seven H11 Agar

(Selective 7H11 Agar)

Composition per 1010.0mL:

Agar .............................................................. ...13.5g

KH2PO4........................................................... ...1.5g

Na2HPO4........................................................... .1.5g

Pancreatic digest of casein................................. .1.0g

NaCl.............................................................. ...0.85g

Monosodium glutamate ................................... ..0.5g

(NH4)2SO4........................................................ ..0.5g

Sodium citrate................................................... ..0.4g

MgSO4*7H2O.................................................. .0.05g

Ferric ammonium citrate................................. ..0.04g

CuSO4*5H2O.................................................. .1.0mg

Pyridoxine..................................................... ..1.0mg

ZnSO4*7H2O ................................................. .1.0mg

Biotin ........................................................... ...0.5mg

CaCl2*2H2O.................................................. ...0.5mg

Malachite Green.......................................... ...0.25mg

Middlebrook OADC enrichment .............. .100.0mL

Antibiotic inhibitor .................................... ...10.0mL

Glycerol ........................................................ .5.0mL

pH 6.6 ± 0.2 at 25oC

Source: This medium is available as a prepared me-
dium from  BD Diagnostic Systems.

Middlebrook OADC Enrichment:

Bovine albumin fraction V .............................. ...5.0g

Glucose ........................................................... ...2.0g

NaCl.............................................................. ...0.85g

Catalase......................................................... ...3.0mg

Oleic acid................................................... ...0.06mL

Preparation of Middlebrook OADC Enrich-
ment: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.

Antibiotic Inhibitor:

Composition per 10.0mL:

Carbenicillin ................................................... ..0.05g

Trimethoprim lactate........................................ ..0.02g

Amphotericin B ............................................. ...0.01g

Polymyxin B............................................. .200,000U

Preparation of Antibiotic Inhibitor: Add com-
ponents to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation  of  Medium:  Add  glycerol  to 900.0mL of distilled/deionized water. Mix thorough-
ly. Add remaining components, except Middlebrook OADC enrichment and antibiotic inhibitor. Mix thor-
oughly. Gently heat. Do not boil. Autoclave for 10 min at 15 psi pressure-121oC. Cool to 50o-55oC. Aseptically add 100.0mL of sterile Middlebrook

OADC enrichment and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour into sterile Petri dish-
es or distribute into sterile tubes.

Use: For the isolation and cultivation of Mycobacte-
rium species from specimens with a mixed flora.


 

SF Broth

(Streptococcus faecalis Broth)

Composition per liter:

Pancreatic digest of casein............................... ..20.0g

Glucose ........................................................... ...5.0g

NaCl.................................................................. ..5.0g

K2HPO4........................................................... ...4.0g

KH2PO4........................................................... ...1.5g

NaN3 ................................................................. .0.5g

Bromcresol Purple ...................................... ...0.032g

pH 6.9 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Auto-
clave for 15 min at 15 psi pressure-121oC.

Use: For the cultivation and differentiation of group
D enterococci (Streptococcus faecalis and Strepto-
coccus faecium) from group D nonenterococci and
from other Streptococcus species. Group D entero-
cocci turn the medium turbid and yellow-brown.


Shigella Broth

Composition per liter:

Pancreatic digest of casein............................... ..20.0g

NaCl.................................................................. ..5.0g

K2HPO4........................................................... ...2.0g

KH2PO4........................................................... ...2.0g

Glucose ........................................................... ...1.0g

Novobiocin solution................................... .. 11.1mL

Tween 80..................................................... ...1.5mL

pH 7.0 ± 0.2 at 25oC

Novobiocin Solution:

Composition per liter:

Novobiocin..................................................... ..0.05g

Preparation of Novobiocin Solution: Add no-
vobiocin to distilled/deionized water and bring vol-
ume to 1.0L. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept novobiocin solution, to distilled/deionized water and bring volume to 988.9mL. Mix thoroughly. Gen-
tly heat and bring to boiling. Autoclave for 15 min at 15 psi pressure-121oC. Cool to 45o-50oC. Aseptical-
ly add sterile novobiocin solution. Mix thoroughly. Aseptically distribute into sterile tubes.

Use: For the isolation and cultivation of Shigella species.


Sierra Medium

Composition per liter:

Agar .............................................................. ...15.0g

Peptone........................................................... ..10.0g

NaCl................................................................. ..5.0g

CaCl2*H2O........................................................ ..0.1g

Tween 80..................................................... .10.0mL

pH 7.4 ± 0.2 at 25oC

Preparation of Medium: Add components, ex-
cept Tween 80, to distilled/deionized water and bring
volume to 990.0mL. Mix thoroughly. Gently heat and
bring to boiling. Autoclave for 15 min at 15 psi pres-
sure-121oC. Cool to 45o-50oC. Separately autoclave
Tween 80 for 15 min at 15 psi pressure-121oC. Cool
to 45o-50oC. Aseptically add 10.0mL of sterile Tween
80. Mix thoroughly. Pour into sterile Petri dishes.

Use: For the differentiation of bacteria based on li-
pase activity. Bacteria with lipase activity form colo-
nies surrounded by a white precipitate.


SIM Medium

Composition per liter:

Peptone........................................................... ..30.0g

Agar ................................................................. ..3.0g

Beef extract...................................................... ...3.0g

Peptonized iron ............................................... ...0.2g

Na2S2O3*5H2O ........................................... ...0.025g

pH 7.3 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.

Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 15.0mL volumes. Autoclave for 15 min at 15 psi pressure-121oC. Allow tubes to cool in an upright position.

Use: For the differentiation of members of the Entero-
bacteriaceae based on H2S production, indole pro-
duction, and motility.


SIM Medium

Composition per liter:

Pancreatic digest of casein............................... ..20.0g

Peptic digest of animal tissue ........................... ..6.1g

Agar ................................................................. ..3.5g

Fe(NH4)2(SO4)2*6H2O ..................................... .0.2g

Na2S2O3*5H2O.................................................. .0.2g

pH 7.3 ± 0.2 at 25oC

Source: This medium is available as a premixed
powder from  BD Diagnostic Systems and Oxoid Un-
ipath.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes in 15.0mL volumes. Autoclave for 15 min at 15 psi pressure-121oC. Allow tubes to cool in an upright position.

Use: For the differentiation of members of the Entero-
bacteriaceae based on H2S production, indole pro-
duction, and motility.


Simmons’ Citrate Agar

(Citrate Agar)

Composition per liter:

Agar .............................................................. ...15.0g

NaCl.................................................................. ..5.0g

Sodium citrate................................................... ..2.0g

K2HPO4........................................................... ...1.0g

(NH4)H2PO4..................................................... ..1.0g

MgSO4*7H2O.................................................. ...0.2g

Bromthymol Blue .......................................... ...0.08g

pH 6.9 ± 0.2 at 25oC

Source: This medium is available as a premixed
powder from  BD Diagnostic Systems and Oxoid Un-
ipath.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat while stirring and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure-121oC. Pour into sterile Petri dishes or leave in tubes.

Use: For the differentiation of Gram-negative bacte-
ria on the basis of citrate utilization. Bacteria that can

utilize citrate as sole carbon source turn the medium

blue.


Skim Milk Agar

Composition per 1100.0mL:

Agar .............................................................. ...15.0g

Pancretic digest of casein................................. ...5.0g

Yeast extract..................................................... ..2.5g

Glucose ........................................................... ...1.0g

Skim milk solution.................................... ..100.0mL

pH 7.0 ± 0.1 at 25oC

Preparation of Skim Milk Solution: Add skim
milk solids to distilled/deionized water and bring
volume to 100.0mL. Mix thoroughly. Autoclave for
15 min at 15 psi pressure-121oC. Cool to 45o-50oC.

Preparation of Medium: Add components, ex-
cept skim milk solution, to distilled/deionized water
and bring volume to 1.0L. Mix thoroughly. Gently
heat and bring to boiling. Distribute into tubes or
flasks. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 45o-50oC. Aseptically add 100.0mL
of cooled, sterile skim milk solution. Mix thoroughly.
Pour into sterile Petri dishes or aseptically distribute
into sterile tubes.

Use: For the cultivation and differentiation of bacte-
ria based on proteolytic activity.


Skirrow Brucella Medium

Composition per liter:

Blood agar base no. 2................................ .940.0mL

Horse blood, lysed defibrinated.................. ..50.0mL

Antibiotic solution ...................................... .10.0mL

pH 7.4 ± 0.2 at 25oC

Blood Agar Base No. 2

Composition per 940.0mL:

Proteose peptone............................................. ..15.0g

Agar .............................................................. ...12.0g

NaCl................................................................. ..5.0g

Yeast extract..................................................... ..5.0g

Liver digest ..................................................... ...2.5g

pH 7.4 ± 0.2 at 25oC

Preparation of Blood Agar Base No. 2: Add
components to distilled/deionized water and bring
volume to 940.0mL. Mix thoroughly. Gently heat
while stirring and bring to boiling. Autoclave for 15
min at 15 psi pressure-121oC. Cool to 45o-50oC.

Antibiotic Solution:

Composition per 10.0mL:

Vancomycin .................................................. ...0.01g

Trimethoprim................................................ ...5.0mg

Polymyxin B ............................................... ...2500U

Preparation of Antibiotic Solution: Add com-

ponents to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: To 940.0mL of sterile cooled blood agar base no. 2, aseptically add 50.0mL of sterile, lysed defibrinated horse blood and 10.0mL of sterile antibiotic solution. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the selective isolation and cultivation of Campylobacter species.


SM ID2 Agar

Composition per liter:

Proprietary

Source: This medium is available from bioM้rieux.

Use: A new chromogenic medium for the selective
isolation and detection of Salmonella, S. typhi, and S.
paratyphi and most Lactose(+) Salmonella present
pale pink to mauve colonies. Other organisms are ei-
ther inhibited, colorless, or pale blue in appearance.


Snyder Agar

Composition per liter:

Glucose ........................................................... .20.0g

Agar .............................................................. ...16.0g

Pancreatic digest of casein............................... ..13.5g

Yeast extract...................................................... ..6.5g

NaCl.................................................................. ..5.0g

Bromcresol Green.......................................... ...0.02g

pH 4.8 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes in 10.0mL volumes. Autoclave
for 15 min at 13 psi pressure-118oC. Do not over-
heat. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and enumeration of lactoba-
cilli in saliva and indication of dental caries activity.


Snyder Test Agar

Composition per liter:

Agar .............................................................. ...20.0g

Glucose ........................................................... .20.0g

Tryptose ........................................................ ...20.0g

NaCl.................................................................. ..5.0g

Bromcresol Green.......................................... ...0.02g

pH 4.8 ± 0.2 at 25oC

Source: This medium is available as a premixed powder from  BD Diagnostic Systems.


Preparation of Medium: Add components to

distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes in 10.0mL volumes. Autoclave
for 15 min at 13 psi pressure-118oC. Do not over-
heat. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and enumeration of lactoba-
cilli in saliva and indication of dental caries activity.


Sodium Chloride Broth, 6.5

Composition per liter:

Beef heart, solids from infusion..................... .500.0g

NaCl.............................................................. ...65.0g

Tryptose ........................................................ ...10.0g

pH 7.4 ± 0.2 at 25oC

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Distribute into tubes or flasks. Auto-
clave for 15 min at 15 psi pressure-121oC.

Use: For the cultivation of enterococci and other salt-tolerant organisms. For the differentiation of mi-
croorganisms based on salt tolerance.


Sodium Chloride Sucrose Medium 900
   (Sodium Chloride SUC Medium 900)

Composition per liter:

Sucrose........................................................... ..97.3g

Pancreatic digest of gelatin............................... .14.5g

NaCl.............................................................. ...14.3g

Agar .............................................................. ...13.3g

Brain heart, solids from infusion .................... ...6.0g

Peptic digest of animal tissue........................... ...6.0g

Yeast extract..................................................... ..5.0g

Glucose ........................................................... ...3.0g

Na2HPO4........................................................... .2.5g

MgSO4 .......................................................... ..0.25g

Horse serum (γ-globulin free,

inactivated 30 min at 56oC)............ ..100.0mL

Carbenicillin solution................................. ...10.0mL

pH 7.4 ± 0.2 at 25oC

Carbenicillin Solution:

Composition per 10.0mL:

Carbenicillin...................................................... ..5.0g

Preparation of Carbenicillin Solution: Add
carbenicillin to distilled/deionized water and bring
volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, ex-
cept carbenicillin solution and horse serum, to dis-
tilled/deionized water and bring volume to 890.0mL.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure-121oC. Cool
to 45o-50oC. Aseptically add carbenicillin solution


 

and horse serum. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of Pseudomonas aerugi-
nosa.


Sodium Chloride Sucrose

Medium 900 with Penicillin G Composition per liter:

Sucrose........................................................... ..97.3g

Pancreatic digest of gelatin............................... .14.5g

NaCl............................................................... ...14.3g

Agar .............................................................. ...13.3g

Brain heart, solids from infusion ..................... ...6.0g

Peptic digest of animal tissue ........................... ..6.0g

Yeast extract...................................................... ..5.0g

Glucose ........................................................... ...3.0g

Na2HPO4........................................................... .2.5g

MgSO4 ........................................................... ..0.25g

Horse serum (γ-globulin free,

inactivated 30 min at 56oC)............ .. 100.0mL

Penicillin solution........................................ .. 10.0mL

pH 7.4 ± 0.2 at 25oC

Penicillin Solution:

Composition per 10.0mL:

Penicillin G............................................. ...500,000U

Preparation of Penicillin Solution: Add peni-
cillin G to distilled/deionized water and bring vol-
ume to 10.0mL. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept penicillin solution and horse serum, to distilled/ deionized water and bring volume to 900.0mL. Mix thoroughly. Gently heat and bring to boiling. Auto-
clave for 15 min at 15 psi pressure-121oC. Cool to 45o-50oC. Aseptically add penicillin solution and horse serum. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use: For the cultivation of Pseudomonas aerugi-
nosa.


Sodium Hippurate Broth

(Hippurate Broth)

Composition per liter:

Beef heart, solids from infusion..................... .500.0g

Tryptose ........................................................ ...10.0g

Sodium hippurate............................................. .10.0g

NaCl.................................................................. ..5.0g

pH 7.4 ± 0.2 at 25oC

Source: Heart infusion broth is available as a pre-
mixed powder from BD Diagnostic Systems.

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.


 

Mix thoroughly. Gently heat and bring to boiling.

Distribute into screw-capped tubes or flasks. Auto-
clave for 15 min at 15 psi pressure-121oC. Tighten caps to prevent drying.

Use: For the identification and differentiation of β-
hemolytic streptococci based on hippurate hydroly-
sis. After inoculation and incubation, tubes are treat-
ed with FeCl3 reagent. A heavy precipitate remaining
after 10-15 min indicates that hippurate has been hy-
drolyzed.


Soft Agar Gelatin Overlay

Composition per plate:

Base agar..................................................... .15.0mL

Soft agar gelatin overlay.............................. ...2.5mL

pH 7.0 ± 0.2 at 25oC

Base Agar:

Composition per liter:

Agar .............................................................. ...15.0g

Peptone.............................................................. .5.0g

NaCl................................................................. ..5.0g

Beef extract...................................................... ...3.0g

MnSO4*H2O.................................................. ...0.05g

Preparation of Base Agar: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Autoclave for 15 min at 15 psi pressure-121oC. Cool
to 45o-50oC.

Soft Agar Gelatin Overlay:

Composition per liter:

Gelatin........................................................... ...15.0g

Agar ................................................................. ..8.0g

Peptone.............................................................. .5.0g

NaCl................................................................. ..5.0g

Beef extract...................................................... ...3.0g

MnSO4*H2O.................................................. ...0.05g

Preparation of Soft Agar Gelatin Overlay:

Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure-121oC. Cool to 45o-50oC.

Preparation  of  Medium:  Aseptically  pour cooled, sterile base agar into sterile Petri dishes in 15.0mL volumes. Allow agar to solidify. Inoculate plates with samples. Overlay each plate with 2.5mL of soft agar gelatin overlay.

Use: For the cultivation and differentiation of micro-
organisms based on proteolytic activity.


Sorbitol MacConkey Agar

(MacConkey Agar with Sorbitol)

  Composition per liter:

Peptone........................................................... ..20.0g 

Agar .............................................................. ...15.0g

Sorbitol ........................................................... .10.0g

NaCl.................................................................. ..5.0g

Bile salts no. 3................................................... ..1.5g

Neutral Red...................................................... .0.03g

Crystal Violet................................................ ...1.0mg

pH 7.1 ± 0.2 at 25oC

Source: This medium is available as a premixed
powder from  BD Diagnostic Systems and Oxoid Un-
ipath.

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.

Use: For the isolation and cultivation of pathogenic Escherichia coli.


Sorbitol MacConkey Agar with BCIG

(SMAC with BCIG)

Composition per liter:

Peptone ........................................................... .20.0g

Agar .............................................................. ...15.0g

Sorbitol ........................................................... .10.0g

NaCl.................................................................. ..5.0g

Proteose peptone................................................ .3.0g

Bile salts mixture.............................................. ...1.5g

5-Bromo-4-chloro-3-indolyl-β-D-glucuronide
       sodium salt... .............................................. .0.1g

Neutral Red...................................................... .0.03g

pH 7.1 ± 0.2 at 25oC

Source: This medium is available from Oxoid.

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat while stirring and bring to boiling. Autoclave for 15 min at 15 psi pressure-
121oC. Pour into sterile Petri dishes.

Use: A selective and differential medium for the de-
tection of Escherichia coli O157 incorporating the chromogen 5-bromo-4-chloro-3-indolyl-β-D-glucu-

ronide (BCIG). The medium combines two different
screening mechanisms for the detection of E. coli
O157, the failure to ferment sorbitol and the absence
of  β-glucuronidase  activity.  The  nonsorbitol-fer-
menting and β-glucuronidase-negative E. coli O157
will appear as straw colored colonies. Organisms
with β-glucuronidase activity will cleave the sub-
strate, leading to a distinct blue-green coloration of
the colonies.


Soy Peptone Broth

Composition per liter:

Papaic digest of soybean meal........................ ..20.0g

NaCl................................................................. ..5.0g

Phenol Red (2% solution).............................. .1.0mL

pH 7.3 ± 0.2 at 25oC

Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Adjust pH to 7.3. Distribute into
tubes or flasks. Autoclave for 15 min at 15 psi pres-
sure-121oC.

Use: For the isolation and cultivation of Mycoplas-
ma species and Ureaplasma species.


Specimen Preservative Medium

Composition per liter:

NaCl................................................................. ..5.0g

Sodium citrate*2H2O......................................... .5.0g

(NH4)2HPO4..................................................... ..4.0g

KH2PO4........................................................... ...2.0g

Yeast extract..................................................... ..1.0g

Sodium deoxycholate....................................... ...0.5g

MgSO4*7H2O.................................................. ...0.4g

Glycerol .................................................. ...300.0mL

pH 7.0 ± 0.2 at 25oC

Preparation of Medium: Add components, ex-
cept glycerol, to distilled/deionized water and bring volume to 700.0mL. Mix thoroughly. Gently heat and bring to boiling. Add 300.0mL of glycerol. Mix thoroughly. Distribute into tubes or flasks. Autoclave for 10 min at 11 psi pressure-116oC.

Use: For the preservation of viable microorganisms
in stool specimens. For the transport of fecal materi-
al.


Spray’s Fermentation Medium

Composition per 1100.0mL:

Neopeptone...................................................... .10.0g

Pancreatic digest of casein.............................. ..10.0g

Agar ................................................................. ..2.0g

Sodium thioglycolate.................................... ..0.025g

Carbohydrate solution.............................. ... 110.0mL

pH 7.4 ± 0.1 at 25oC

Carbohydrate Solution:

Composition per 200.0mL:

Carbohydrate.................................................. ..20.0g

Preparation of Carbohydrate Solution: Add carbohydrate to distilled/deionized water and bring volume to 200.0mL. Glucose or glycerol may be used. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, ex-
cept agar and carbohydrate solution, to distilled/

deionized water and bring volume to 990.0mL. Mix
thoroughly. Adjust pH to 7.4. Add agar. Gently heat
and bring to boiling. Distribute into tubes in 9.0mL
volumes. Autoclave for 15 min at 15 psi pressure-
121oC. Cool to 25oC. Immediately prior to use heat
tubes in a boiling water bath for 10 min. Cool to
45oC. Aseptically add 1.0mL of sterile carbohydrate
solution. Mix thoroughly.

Use:  For  the  cultivation  and  differentiation  of Clostridium perfringens based on carbohydrate fer-
mentation patterns.