M Endo Broth MF: Intended Use Formula

m Endo Broth MF™
Formula
Intended Use Difco™ m Endo Broth MF™
m Endo Broth MF™* is used for enumerating coliform organisms Approximate Formula* Per Liter
in water by membrane filtration. Yeast Extract ............................................................... 1.5 g
*MF is a trademark of Millipore Corporation. Casitone...................................................................... 5.0 g
Thiopeptone................................................................ 5.0 g
Tryptose..................................................................... 10.0 g
Summary and Explanation Lactose...................................................................... 12.5 g
m Endo Broth MF is prepared according to the formulation Sodium Desoxycholate................................................. 0.1 g
of the Millipore Corporation1 for selectively isolating coliform Dipotassium Phosphate................................................ 4.375 g
Monopotassium Phosphate.......................................... 1.375 g
bacteria from water and other specimens using the membrane Sodium Chloride.......................................................... 5.0 g
filtration technique. The medium is a combination of the former Sodium Lauryl Sulfate................................................... 0.05 g
m HD Endo Medium and Lauryl Tryptose Broth. Sodium Sulfite.............................................................. 2.1 g
Basic Fuchsin................................................................ 1.05 g
The American Public Health Association (APHA) specifies *Adjusted and/or supplemented as required to meet performance criteria.
using m Endo Broth MF in the standard total coliform

membrane filtration procedure for testing water2 and bottled Directions for Preparation from
water.3 APHA also specifies using m Endo Broth MF in the Dehydrated Product
delayed-incubation total coliform procedure by adding sodium 1. Dissolve 48 g of the powder in 1 L of purified water
benzoate to make m Endo preservative medium.2 The coliform containing 20 mL nondenatured ethanol.
bacteria are defined as bacteria that produce a red colony with 2. Heat to boiling but avoid over-heating. DO NOT
a metallic sheen within 24 hours incubation at 35°C on an AUTOCLAVE.
Endo-type medium. 3. Test samples of the finished product for performance using
stable, typical control cultures.
Principles of the Procedure
CAUTION: Protect from light. Use immediately.
m Endo Broth MF contains peptones as sources of carbon,
nitrogen, vitamins and minerals. Yeast extract supplies
Procedure
B-complex vitamins, which stimulate bacterial growth. 1. Place a membrane filter absorbent pad inside a sterile
Lactose is the carbohydrate. Phosphates are buffering agents. 60 mm Petri dish.
Sodium chloride maintains the osmotic balance of the medium. 2. Add 1.8-2.0 mL m Endo Broth MF to each pad.
Sodium desoxycholate and sodium lauryl sulfate are added 3. Filter the water sample through a membrane filter.
as inhibitors of gram-positive bacteria. Basic fuchsin is a pH 4. Place filter top side up on the pad using a rolling motion to
indicator. Sodium sulfite is added to decolorize the basic fuchsin avoid entrapping air bubbles.
solution. The ethanol additive increases the antibacterial 5. Invert the dish and incubate for 22-24 hours at 35 ± 0.5°C.
nature of the formulation. 6. Observe and count all colonies that are red and have a
Lactose-fermenting bacteria produce acetaldehyde that reacts metallic sheen.
with the sodium sulfite and fuchsin to form red colonies. The
development of a metallic sheen occurs when the organism Expected Results
produces aldehydes with the rapid fermentation of lactose. All colonies that are red and have the characteristic metallic
If the inoculum is too heavy, the sheen will be suppressed. sheen are considered coliforms. The sheen may cover the
Lactose-nonfermenting bacteria form clear, colorless colonies. entire colony, may only be in the center or may appear only
around the edges.
Limitations of the Procedure

Occasionally, noncoliform organisms may produce typical
sheen colonies. Coliform organisms may also occasionally
produce atypical colonies (dark red or nucleated colonies
without sheen). It is advisable to verify both colony types.2
Difco™ & BBL™ Manual, 2nd Edition

User Quality Control References
1. Fifield and Schaufus. 1958. J. Am. Water Works Assoc. 50:193.
Identity Specifications 2. 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.
Difco™ m Endo Broth MF™ 3. Kim and Feng. 2001. In Downes and Ito (ed.), Compendium of methods for the microbiological
examination of foods, 4th ed. American Public Health Association, Washington, D.C.
Dehydrated Appearance: Pinkish purple, free-flowing, homogeneous.
Solution: 4.8% solution, partially soluble in purified water
containing 2% ethanol upon boiling. Solution is Availability
pinkish-red, opalescent with precipitate. Difco™ m Endo Broth MF™
Prepared Appearance: Pinkish-red, opalescent with precipitate. COMPF SMD SMWW
Reaction of 4.8% Cat. No. 274920 Dehydrated – 100 g
Solution at 25°C: pH 7.2 ± 0.1 274930 Dehydrated – 500 g
Cultural Response
Difco™ m Endo Broth MF™
Prepare the medium per label directions. Use the membrane filter technique
to inoculate filters. Incubate on pads saturated with m Endo Broth MF at
35 ± 2°C for 24 ± 2 hours.
Inoculum COLONY
Organism ATCC™ CFU RECOVERY COLOR
Escherichia coli 25922 20-80 Good Red with green
metallic sheen
Salmonella enterica
subsp. enterica Colorless
serotype Typhimurium 14028 20-80 Good to pink
Staphylococcus Marked
aureus 25923 103-2×103 inhibition –
Escherichia coli Salmonella

ATCC™ 25922 Typhimurium
ATCC™ 14028
Difco™ & BBL™ Manual, 2nd Edition

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m Endo Broth MF™

using m Endo Broth MF in the standard total coliform

Limitations of the Procedure

Difco™ & BBL™ Manual, 2nd Edition

Escherichia coli Salmonella

Difco™ & BBL™ Manual, 2nd Edition

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