EC Medium with MUG

References Availability
1. Hajna and Perry. 1943. Am. J. Public Health 33:550.
2. Tennant, Reid, Rockwell and Bynoe. 1961. Can. J. Microbiol. 1:733.
Difco™ EC Medium
3. Fishbein and Surkiewicz. 1964. Appl. Microbiol. 12:127. AOAC BAM CCAM COMPF EPA ISO SMD SMWW
4. Eaton, Rice and Baird (ed.). 2005. Standard methods for the examination of water and wastewater,
21st ed., online. American Public Health Association, Washington, D.C. Cat No. 231420 Dehydrated – 100 g
5. Downes and Ito (ed.). 2001. Compendium of methods for the microbiological examination of foods, 231430 Dehydrated – 500 g
4th ed. American Public Health Association, Washington, D.C.
6. Wehr and Frank (ed.). 2004. Standard methods for the examination of dairy product, online. American
231410 Dehydrated – 10 kg
Public Health Association, Washington, D.C.
7. U.S. Food and Drug Administration. 2001. Bacteriological analytical manual, online. AOAC Interna-
tional, Gaithersburg, Md.
8. Horwitz (ed.). 2007. Official methods of analysis of AOAC International, 18th ed., online. AOAC
International, Gaithersburg, Md.
9. Ray. 1986. J. Food. Prot. 49:651.

EC Medium with MUG

Intended Use
EC Medium with MUG is used for detecting Escherichia coli
in water, food and milk.
Summary and Explanation
EC Medium was developed by Hajna and Perry1 to improve
the methods for the detection of coliforms and E. coli. This
medium consists of a buffered lactose broth with the addition
of 0.15% Bile Salts No. 3. Growth of sporeformers and fe-
cal streptococci is inhibited by the bile salts, while growth of
E. coli is enhanced. EC Medium with MUG is the same formula
as EC Medium with the addition of 4 methyl-umbelliferyl-
β-D-glucuronide.
Feng and Hartman2 developed a rapid assay for E. coli by
incorporating 4-methylumbelliferyl-β-D-glucuronide (MUG)
into Lauryl Tryptose Broth at a final concentration of
100 µg/mL. Robison3 compared the fluorogenic assay with
present methodology and found that total agreement between
the two methods was 94.8%. Moburg4 determined the amount
of MUG could be reduced to a final concentration of 50 µg/mL
without adversely affecting results. Koburger and Miller5
recommended the incorporation of MUG into EC Broth for use
in testing shellfish.
EC Medium with MUG is prepared according to the formula
specified by the U.S. Environmental Protection Agency6 and
standard methods for water and food testing.7,8
Principles of the Procedure
Peptone provides the nitrogen, vitamins and amino acids in
EC Medium with MUG. Lactose is the carbon source in this
medium. Bile Salts No. 3 is the selective agent against gram- E
positive bacteria, particularly bacilli and fecal streptococci.
Dipotassium phosphate and monopotassium phosphate are
buffering agents. Sodium chloride maintains the osmotic balance
of the medium.
E. coli produces the enzyme glucuronidase that hydrolyzes MUG
to yield a fluorogenic product that is detectable under long
wave (366 nm) UV light. The addition of MUG to EC Medium
provides another criterion, in addition to growth response and
gas production, to determine the presence of E. coli in food and
environmental samples.

User Quality Control

Identity Specifications
Difco™ EC Medium with MUG
Dehydrated Appearance: Light beige, free-flowing, homogeneous.
Solution: 3.71% solution, soluble in purified water. Solution is light amber, clear.
Prepared Appearance: Light amber, clear.
Reaction of 3.71%
Solution at 25°C: pH 6.9 ± 0.2

Cultural Response
Difco™ EC Medium with MUG
Prepare the medium per label directions. Inoculate tubes in duplicate with fresh 18-24 hour cultures. Incubate
the first set at 35 ± 2°C for 24 ± 2 hours and the second set at 44.5 ± 0.2°C for 24 ± 2 hours. Read fluorescence
under a long-wave UV light.
RECOVERY at RECOVERY at
Organism ATCC™ 35°C/Gas 44.5°C/Gas Fluorescence
Enterobacter aerogenes 13048 Good/± Inhibition to good/– –
Enterococcus faecalis 19433 Inhibition/– Inhibition to good/– –
Escherichia coli 25922 Good/+ Good/+ + Escherichia coli
ATCC™ 25922

201

Difco Manual Sect III E.indd 201 3/16/09 4:14:02 PM

 Section III
E EC Medium with MUG, cont.
Formula
Difco™ EC Medium with MUG
Approximate Formula* Per Liter
Tryptose..................................................................... 20.0 g
Lactose........................................................................ 5.0 g
Bile Salts No. 3............................................................. 1.5 g
Dipotassium Phosphate................................................ 4.0 g
Monopotassium Phosphate.......................................... 1.5 g
Sodium Chloride.......................................................... 5.0 g
MUG (4-methylumbelliferyl-β-D-glucuronide)............... 0.05 g
*Adjusted and/or supplemented as required to meet performance criteria.
Directions for Preparation from
Dehydrated Product
1. Dissolve 37.1 g of the powder in 1 L of purified water.
Mix thoroughly.
2. Warm slightly to completely dissolve the powder.
3. Dispense into test tubes containing inverted fermentation
vials.
4. Autoclave at 121°C for 15 minutes.
5. Test samples of the finished product for performance using
stable, typical control cultures.
Procedure
Follow the methods and procedures as stated in appropriate
references.6-8
Expected Results
Following incubation, observe tubes for growth, production of
gas and fluorescence. Positive gas production is demonstrated
by displacement of the medium from the fermentation vial.
Positive MUG reactions exhibit a bluish fluorescence under
long-wave (approximately 366 nm) UV light. Typical strains
of E. coli are positive for both gas production and fluorescence.
Non-E. coli coliforms that grow may exhibit fluorescence but
will not produce gas.
EC Medium, Modified
Novobiocin Antimicrobic Supplement
Intended Use
EC Medium, Modified is used with Novobiocin Antimicrobic
Supplement in the detection of Escherichia coli O157:H7 in
meat and poultry products.
Summary and Explanation
EC Medium, Modified and Novobiocin Antimicrobic Supple-
ment are based on the formula for modified EC broth with
novobiocin (mEC+n) as described by Okrend and Rose.1 In
modifying the EC Medium formula, Okrend and Rose reduced
the Bile Salts No. 3 from 1.5 g per liter to 1.12 g per liter and
added 20 mg per liter of sodium novobiocin. Okrend et al.
reported that mEC+n was useful in the enrichment and detection
of E. coli O157:H7 from meats and poultry products.2-4
202
Difco Manual Sect III E.indd 202
Strains of Salmonella, Shigella and Yersinia that produce
glucuronidase may be encountered. These strains must be
distinguished from E. coli on the basis of other parameters; i.e.,
gas production, growth at 44.5°C.
Limitations of the Procedure
1. Strains of E. coli that fail to grow in EC Medium with MUG,
fail to produce gas, or fail to produce glucuronidase may
infrequently be encountered.
2. The presence of endogenous glucuronidase in shellfish
samples may cause false positive fluorescent reactions at
the presumptive stage. To prevent this problem, the use of
EC Medium with MUG in the confirmatory stage has been
recommended.5
References
1. Hajna and Perry. 1943. Am. J. Public Health 33:550.
2. Feng and Hartman. 1982. Appl. Environ. Microbiol. 43:1320.
3. Robison. 1984. App. Environ. Microbiol. 48:285.
4. Moberg. 1985. Appl. Environ. Microbiol. 50:1383.
5.
6.
Koburger and Miller. 1985. J. Food Prot. 48:244.
Federal Register. 1991. National primary drinking water regulation; analytical techniques; coliform
bacteria. Fed. Regist. 56:636.
7. Eaton, Rice and Baird (ed.). 2005. Standard methods for the examination of water and wastewater,
21st ed., online. American Public Health Association, Washington, D.C.
8. U.S. Food and Drug Administration. 2001. Bacteriological analytical manual, online. AOAC Interna-
tional, Gaithersburg, Md.
Availability
Difco™ EC Medium with MUG
BAM CCAM EPA SMWW
Cat. No. 222100 Dehydrated – 100 g
222200 Dehydrated – 500 g
Principles of the Procedure
Peptone supports good growth of E. coli O157:H7 and is rich
in peptides and nitrogen. Lactose is an additional source of
carbon for organisms, such as E. coli, that can ferment this
sugar. Dipotassium phosphate and monopotassium phosphate
are buffers that facilitate recovery of injured cells. Sodium
chloride provides a suitable ionic environment for growth of
microorganisms.
Selectivity of the medium is achieved by the incorporation of Bile
Salts No. 3 into the base medium and by the addition of sodium
novobiocin to the complete medium. These agents suppress
the growth of nuisance organisms commonly found in foods.
The sodium novobiocin is provided in the freeze-dried state
as Novobiocin Antimicrobic Supplement. This supplement is
rehydrated before use with sterile purified water.
3/16/09 4:14:04 PM

Lauryl Tryptose Broth • Lauryl Sulfate Broth
Intended Use
Lauryl Tryptose Broth and Lauryl Sulfate Broth, which are
also known as Lauryl Sulfate Tryptose (LST) Broth, are used
for the detection of coliform organisms in materials of sanitary
importance.
Summary and Explanation
Mallmann and Darby developed this medium for the detection
of coliform organisms by American Public Health Association
(APHA) procedures.1 They incorporated sodium lauryl sulfate
into the formulation since it proved to be selective but not
inhibitory for coliforms.
This medium is used for the detection of coliforms in foods 2
and dairy products.3 It is now the medium of choice for use
in the presumptive phase of the Standard Total Coliform
Multiple-Tube (MPN) Test for the microbiological examination
of water.4 It is also listed in the Official Methods of Analysis of
AOAC International.5
Principles of the Procedure
Peptone provides essential growth substances, such as nitrogen
and carbon compounds, sulfur and trace ingredients. The
potassium phosphates provide buffering capacity. Sodium
chloride maintains osmotic equilibrium.
User Quality Control
NOTE: Differences in the Identity Specifications and Cultural Response testing for media offered as both Difco™ and BBL™ brands may reflect differences in the
development and testing of media for industrial and clinical applications, per the referenced publications.
Identity Specifications
Difco™ Lauryl Tryptose Broth
Dehydrated Appearance: Light beige, free-flowing, homogeneous.
Solution: 3.56% solution, soluble in purified water upon
warming. Solution is light to medium amber,
clear to very slightly opalescent.
Prepared Appearance: Light to medium amber, clear to very slightly
opalescent.
Reaction of 3.56%
Solution at 25°C: pH 6.8 ± 0.2
Cultural Response
Difco™ Lauryl Tryptose Broth
Prepare the medium per label directions. Inoculate and incubate at
35 ± 2°C for 24 ± 2 hours or longer, if necessary.
INOCULUM
ORGANISM ATCC™ CFU RECOVERY GAS
Enterobacter aerogenes 13048 30-300 Good +*
Escherichia coli 25922 30-300 Good +
Salmonella enterica
subsp. enterica
serotype Typhimurium 14028 30-300 Good –
Staphylococcus aureus 25923 3×10 -10 Marked to
2 3
– Proteus mirabilis 12453 103-104 Good –
complete inhibition
*Gas production positive within 48 ± 3 hours.
Difco Manual Sect III L.indd 293
Lauryl Tryptose Broth
Lactose provides a source of fermentable carbohydrate
for coliform organisms. The fermentation of lactose with gas
formation is a presumptive test for coliforms. Sodium lauryl
sulfate inhibits organisms other than coliforms.
Formulae
Difco™ Lauryl Tryptose Broth
Approximate Formula* Per Liter
Tryptose..................................................................... 20.0 g
Lactose........................................................................ 5.0 g
Dipotassium Phosphate................................................ 2.75 g
Monopotassium Phosphate.......................................... 2.75 g
Sodium Chloride.......................................................... 5.0 g
Sodium Lauryl Sulfate................................................... 0.1 g
BBL™ Lauryl Sulfate Broth
Approximate Formula* Per Liter
Pancreatic Digest of Casein........................................ 20.0 g
Lactose........................................................................ 5.0 g
Dipotassium Phosphate................................................ 2.75 g
Monopotassium Phosphate.......................................... 2.75 g
Sodium Chloride.......................................................... 5.0 g
Sodium Lauryl Sulfate................................................... 0.1 g
*Adjusted and/or supplemented as required to meet performance criteria.
Identity Specifications
BBL™ Lauryl Sulfate Broth
Dehydrated Appearance: Fine, homogeneous, free of extraneous
material.
Solution: 3.56% solution, soluble in purified water.
Solution is pale to light, tan to yellow, clear
to slightly hazy.
Prepared Appearance: Pale to light, tan to yellow, clear to slightly
Reaction of 3.56%
hazy.
L
Solution at 25°C: pH 6.8 ± 0.2
Cultural Response
BBL™ Lauryl Sulfate Broth
Prepare the medium per label directions. Inoculate and incubate at
35 ± 2°C for 48 hours.
INOCULUM
ORGANISM ATCC™ CFU REcovery gas
Enterobacter aerogenes 13048 103-104 Good +
Enterococcus faecalis 29212 104-105 Partial to –
complete inhibition
Escherichia coli 25922 103-104 Good +
293
3/16/09 4:35:26 PM
 Section III
L Lauryl Tryptose Broth, cont.
Directions for Preparation from
Dehydrated Product
Difco™ Lauryl Tryptose Broth
1. Suspend 35.6 g of the powder in 1 L of purified water. Mix
thoroughly.
2. Warm slightly to completely dissolve the powder.
3. Dispense required amounts into tubes containing inverted
fermentation vials (see table).
4. Autoclave at 121°C for 15 minutes. Cool the broth as quickly
as possible.
5. Test samples of the finished product for performance using
stable, typical control cultures.
BBL™ Lauryl Sulfate Broth
1. Suspend 35.6 g of the powder in 1 L of purified water.
2. Dispense in test tubes, containing inverted Durham tubes,
in 10 mL amounts for testing samples of 1 mL or less. For
testing 10 mL quantities of samples, dissolve 71.2 g of the
powder in 1 L of purified water and distribute in 10 mL
amounts. The concentration of the medium should be
varied according to the size of the test samples (see table).
3. Autoclave at 121°C for 15 minutes. After autoclaving, cool
the broth as quickly as possible.
4. Test samples of the finished product for performance using
stable, typical control cultures.
Preparation of Lauryl Tryptose (Sulfate) Broth4
amount of dehydrated
medium in volume of medium
Inoculum mL the tube mL medium+inoculum mL required g/L
1 10 or more 11 or more 35.6
10 10 20 71.2
10 20 30 53.4
20 10 30 106.8
100 50 150 106.8
100 35 135 137.1
100 20 120 213.6
Lauryl Tryptose Broth with MUG
Lauryl Sulfate Broth with MUG
Intended Use
Lauryl Tryptose Broth with MUG and Lauryl Sulfate Broth
with MUG, which are also known as Lauryl Sulfate Tryptose
Broth with MUG (LST-MUG), are used for the detection
of Escherichia coli in water, food and dairy samples by a
fluorogenic procedure.
Summary and Explanation
E. coli is used as an indicator organism of unsanitary conditions.
A number of selective media are recommended for use in enrich-
294
Difco Manual Sect III L.indd 294
NOTE: Refrigerated broth generally becomes cloudy or forms
precipitates but clears upon warming to room temperature.
However, clarity is not important because only gas production
is significant.
Procedure
Refer to the official test procedures for the detection of coliforms
in the compendia of methods for microbiological examination
of foods, dairy products and waters.2-5
Expected Results
After incubation of the tubes with loosened caps at 35 ± 0.5°C
for 24 hours, examine for turbidity and for gas production in
the Durham fermentation tubes. If no gas has formed and been
trapped in the inverted tube, reincubate and reexamine after
48 hours.2-5
Turbidity of the medium accompanied by formation of gas in
any amount in the Durham tubes within 48 hours is a positive
presumptive test for the presence of coliforms in the sample.2-5
The result should be confirmed by additional standard testing.
References
1. Mallmann and Darby. 1941. Am. J. Public Health 31:127.
2. Downes and Ito. 2001. Compendium of methods for the microbiological examination of foods. 4th
ed. American Public Health Association, Washington, D.C.
3. Wehr and Frank (ed.). 2004. Standard methods for the examination of dairy products, 17th ed.
American Public Health Association. Washington, D.C.
4. Eaton, Rice and Baird (ed.). 2005. Standard methods for the examination of water and wastewater,
21st ed., online. American Public Health Association, Washington, D.C.
5. Horwitz (ed.). 2007. Official methods of analysis of AOAC International. 18th ed., online. AOAC
International. Gaithersburg, Md.
Availability
Difco™ Lauryl Tryptose Broth
AOAC BAM CCAM COMPF ISO SMD SMWW
Cat. No. 224140 Dehydrated – 100 g
224150 Dehydrated – 500 g
224120 Dehydrated – 2 kg
224130 Dehydrated – 10 kg
BBL™ Lauryl Sulfate Broth
AOAC BAM CCAM COMPF EPA ISO SMD SMWW
Cat. No. 211338 Dehydrated – 500 g
211339 Dehydrated – 5 lb (2.3 kg)
294369 Dehydrated – 25 lb (11.3 kg)
ment, presumptive identification and confirmatory procedures
for demonstrating the presence of coliforms in material of
sanitary importance. These procedures require the incubation
of test media for up to 7 days.
The presence of the fluorogenic compound, MUG (4-
methylumbelliferyl-β-D-glucuronide), in this medium permits
the rapid detection of E. coli when the medium is observed for
fluorescence using a long-wave UV light source, and further
confirmation is not required.1,2 MUG detects anaerogenic strains
which may not be detected in the conventional procedure.1
3/16/09 4:35:28 PM

User Quality Control
NOTE: Differences in the Identity Specifications and Cultural Response testing for media offered as both Difco™ and BBL™ brands may reflect differences in the
development and testing of media for industrial and clinical applications, per the referenced publications.
Identity Specifications
Difco™ Lauryl Tryptose Broth with MUG
Dehydrated Appearance: Light beige, free flowing, homogeneous.
Solution: 3.57% solution, soluble in purified water upon
warming. Solution is light to medium amber,
clear to very slightly opalescent.
Prepared Appearance: Light to medium amber, clear to very slightly
opalescent.
Reaction of 3.57%
Solution at 25°C: pH 6.8 ± 0.2
Cultural Response
Difco™ Lauryl Tryptose Broth with MUG
Prepare the medium per label directions. Inoculate and incubate at
35 ± 2°C for 24 ± 2 hours or longer, if necessary.
INOCULUM FLUO-
ORGANISM ATCC™ CFU RECOVERY GAS RESCENCE
Enterobacter
aerogenes 13048 30-100 Good +* –
Escherichia coli 25922 30-100 Good + +
Salmonella enterica
subsp. enterica
serotype Typhimurium 14028 30-100 Good – –
Staphylococcus 25923 3×10 -10 Marked –
2 3
– Escherichia coli 25922 103-104 Good + +
aureus to complete
inhibition
*Gas production positive within 48 ± 3 hours.
Feng and Hartman, using a MUG-containing medium in
microtitration plates, reported β-glucuronidase activity in 96%
of E. coli, 100% of enterotoxigenic E. coli, 17% of Salmonella
spp., and 40% of Shigella spp., while all other genera tested
were negative; most reactions occurred within 4 hours, but
some weakly β-glucuronidase-positive strains required overnight
incubation.1 In the presence of large numbers of Proteus
vulgaris, which may suppress gas production by E. coli, fluores-
cence due to E. coli was detected within 15 hours.1
In a comparison, with conventional methods, Robison reported
94.8% agreement, a false-positive rate of 4.8%, attributable to
the presence of streptococci in the samples, and no false-nega-
tives.2
These media are included in the compendia of methods for
the detection and enumeration of coliform organisms in food 3
and dairy 4 products and in the Official Methods of Analysis of
AOAC International.5
Principles of the Procedure
Lactose is a source of energy for organisms. Peptone provides
additional nutrients. The phosphate compounds provide
buffering capacity. Sodium lauryl sulfate is inhibitory to many
organisms but not for coliforms.
Difco Manual Sect III L.indd 295
Lauryl Tryptose Broth with MUG, cont.
Identity Specifications
BBL™ Lauryl Sulfate Broth with MUG
Dehydrated Appearance: Fine, homogeneous, free of extraneous
material.
Solution: 3.57% solution, soluble in purified water.
Solution is pale to light, tan to yellow, clear
to slightly hazy.
Prepared Appearance: Pale to light, tan to yellow, clear to slightly
hazy.
Reaction of 3.57%
Solution at 25°C: pH 6.8 ± 0.2
Cultural Response
BBL™ Lauryl Sulfate Broth with MUG
Prepare the medium per label directions. Inoculate and incubate at
35 ± 2°C for 48 hours.
INOCULUM fluo-
ORGANISM ATCC™ CFU recovery gas Rescence
Enterobacter
aerogenes 13048 103-104 Good + –
Enterococcus 29212 104-105 Partial to – –
faecalis complete
inhibition
The substrate 4-methylumbelliferyl-β-D-glucuronide is
hydrolyzed by an enzyme, β-glucuronidase, possessed by most
E. coli and a few strains of Salmonella, Shigella and Yersinia,
to yield a fluorescent end product, 4-methylumbelliferone.1,2
Development of fluorescence permits the detection of E. coli
in pure or mixed cultures within 4-24 hours following inocula-
tion and incubation of the medium.
Formulae
Difco™ Lauryl Tryptose Broth with MUG
Approximate Formula* Per Liter
Tryptose..................................................................... 20.0 g L
Lactose........................................................................ 5.0 g
Dipotassium Phosphate................................................ 2.75 g
Monopotassium Phosphate.......................................... 2.75 g
Sodium Chloride.......................................................... 5.0 g
Sodium Lauryl Sulfate................................................... 0.1 g
MUG (4-methylumbelliferyl-β-D-glucuronide)............... 0.05 g
BBL™ Lauryl Sulfate Broth with MUG
Approximate Formula* Per Liter
Pancreatic Digest of Casein........................................ 20.0 g
Lactose........................................................................ 5.0 g
Dipotassium Phosphate................................................ 2.75 g
Monopotassium Phosphate.......................................... 2.75 g
Sodium Chloride.......................................................... 5.0 g
Sodium Lauryl Sulfate................................................... 0.1 g
4-Methylumbelliferyl-β-D-glucuronide ......................... 0.05 g
*Adjusted and/or supplemented as required to meet performance criteria.
295
3/16/09 4:35:29 PM
 Section III
L Lauryl Tryptose Broth with MUG, cont.
Directions for Preparation from
Dehydrated Product
Difco Lauryl Tryptose Broth with MUG
™ Observe the medium periodically during the incubation
1. Suspend 35.7 g of the powder in 1 L of purified water and
warm slightly to dissolve completely.
2. Dispense into test tubes containing inverted fermentation
vials.
3. Autoclave at 121°C for 15 minutes.
4. Test samples of the finished product for performance using
stable, typical control cultures.
BBL™ Lauryl Sulfate Broth with MUG
1. Dissolve 35.7 g of the powder in 1 L of purified water.
2. Dispense in test tubes, containing inverted Durham tubes,
in 10 mL amounts for testing samples of 1 mL or less. For
testing 10 mL quantities of samples, dissolve 71.4 g of the
powder in 1 L of purified water and distribute in 10 mL
amounts. The concentration of the medium should be
varied according to the size of the test samples.
3. Autoclave at 121°C, preferably for 12 minutes, but not
exceeding 15 minutes. After autoclaving, cool the broth as
quickly as possible.
4. Test samples of the finished product for performance using
stable, typical control cultures.
NOTE: Refrigerated broth generally becomes cloudy or forms
precipitates but clears upon warming to room temperature.
However, clarity is not important because only gas production
is significant.
Lecithin Lactose Agar
Intended Use
Lecithin Lactose Agar is recommended for the isolation and
differentiation of histotoxic clostridia from clinical specimens.1
It is particularly useful in differentiating Clostridium perfringens,
C. sordellii, C. novyi, C. septicum and C. histolyticum.
Summary and Explanation
Culture media containing egg yolks were useful in isolating,
cultivating and identifying species of histotoxic clostridia.
In 1948, McClung and Toabe reported on the use of egg yolk
medium for this purpose.1 Willis and Hobbs added milk and
lactose to the egg yolk in a formulation designed to group
clostridia on the basis of production of lecithinase, hydrolysis
of casein and lactose fermentation.2 Neomycin sulfate was
included in their formulation in order to make it a selective
medium.
Willis, in response to problems in the obtaining and processing
of antibiotic-free eggs, developed an egg-free medium in which
purified lecithin was substituted for the egg yolk.3 In order to
further refine the growth-promoting and selective properties,
Ellner and O’Donnell developed the formulation, designated as
Lecithin Lactose Agar, in which a decreased concentration of
neomycin was employed and to which sodium azide was added.4
296
Difco Manual Sect III L.indd 296
Procedure
Follow standard methods for the test being performed.3-5
period for the development of fluorescence, using a long-wave
UV light source (approximately 366 nm) as well as for charac-
teristic growth and/or gas production.
Expected Results
The presence of E. coli is detected by the appearance of fluores-
cence throughout the tube.
References
1. Feng and Hartman. 1982. Appl. Environ. Microbiol. 43:1320.
2. Robison. 1984. Appl. Environ. Microbiol. 48:285.
3. Downes and Ito. 2001. Compendium of methods for the microbiological examination of foods, 4th
ed. American Public Health Association, Washington, D.C.
4. Wehr and Frank (ed.). 2004. Standard methods for the examination of dairy products, 17th ed.
American Public Health Association, Washington, D.C.
5. Horwitz (ed.). 2007. Official methods of analysis of AOAC International, 18th ed., online. AOAC
International, Gaithersburg, Md.
Availability
Difco™ Lauryl Tryptose Broth with MUG
AOAC BAM COMPF SMD SMWW
Cat. No. 211740 Dehydrated –100 g
211744 Dehydrated – 500 g
BBL™ Lauryl Sulfate Broth with MUG
AOAC BAM COMPF SMD SMWW
Cat. No. 298076 Dehydrated –500 g
This medium continues to be an important one for visualizing
and characterizing the histotoxic clostridia.
Principles of the Procedure
Lecithin Lactose Agar provides differentiation of clostridia
by the demonstration of lecithinase production and lactose
fermentation. The medium is rendered selective by the addition
of neomycin and sodium azide. Gram-negative and aerobic
gram-positive rods are inhibited. Growth of gram-positive cocci
is suppressed.
On this medium, the production of a zone of opalescence
surrounding colonies indicates lecithinase production. A yellow
color surrounding colonies indicates lactose fermentation
due to the effect of the lowered pH on the bromcresol purple
indicator.
Procedure
As soon as possible after receipt in the laboratory, inoculate
the specimen onto a reduced Lecithin Lactose Agar plate and
streak for isolation. Since some anaerobes may be inhibited due
to the selective nature of the medium, it is advisable to include a
nonselective medium such as CDC Anaerobe 5% Sheep Blood
Agar.
3/16/09 4:35:31 PM