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Lactose BrothArt. No. 02-105

References

 APHA-AWWA-WPCF (1998) Standard methods or the examination o

water and wastewater. 20th ed. APHA Washington

DOWNES, F.P. & K. ITO (2001) Compendium o Methods or theMicrobiological Examination o Foods. 4th ed. APHA. Washington.

FDA (Food and Drug Adminstrations) (1998) Bacteriological Analytical

Manual 8th ed. Rev A. AOAC International. Gaithersburg. VA. USA.

ISO 9308-2 Standard. (1990) Water Quality - Detection and enumeration

o coliorm organisms, thermotolerant coliorm and presumptive E. coli

- MPN technique.

ISO/TS 11133-1: 2009. Microbiology o ood and animal eeding stus.-

Guidelines on preparation and production o culture media. Part 1:

General guidelines on quality assurance or the preparation o culture

media in the laboratory.

ISO/TS 11133-2: 2003 Corr. 2004. Microbiology o ood and animal

eeding stus.- Guidelines on preparation and production o culture

media. Part 2: Practical guidelines on perormance testing o culturemedia.

ISO 21150:2006 Standard. Cosmetics - Detection o Escherichia coli.

US PHARMACOPOEIA (2005) <61> Microbial limit test. US

Pharmacopeial Conv. Inc. Rockville. MD. USA.

VANDERZANT & SPLITTSTOESSER (1992) Compendium o Methods or

the Microbiological Examination o Foods. 3rd ed. APHA. Washington.

Storage

For laboratory use only. Keep tightly closed, away rom bright light, in a

cool dry place (+4ºC to 30ºC and <60% RH).

Specification

Medium or pre-enrichment and detection o enterobacteria and

coliorms in milk and water according to ISO standards.

Formula* in g/L

Peptone 5,00

Meat extract 3,00

Lactose 5,00

Final pH 6,9 ± 0,2 at 25ºC

* Adjusted and /or supplemented as required to meet perormance criteria

Directions

 Add 13 g o powder to 1 L o distilled water, or in the quantity required or

the desired concentration.

Dissolve and distribute into containers ftted with Durham tubes. Sterilize

in the autoclave at 121ºC or 15 minutes. Avoid further reheating.

Description

Lactose Broth is a classical medium or use in the presumptive testing or

coliorms and or the enrichment o Salmonella. This ormulation is per the

standards recommended by APHA, AWWA, USP-NF and ISO.

It is commonly used with Durham ermentation tubes or the detection

o gas ormation. I a specifc volume o sample is to be inoculated this

must be taken into consideration when making up the medium as the

concentration must not be alteredon addition o the inoculum.

  Although it is not Eijkman’s original ormulation, this broth providesexcellent results in assays o gas production at 45ºC, which is a

characteristic o Escherichia coli.

While preparing this medium it is important to avoid overheating and to

distribute it into tubes beore sterilization.

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Quality control

Incubation temperature: 35°C ± 2,0

Incubation time: 24 - 48 h

Inoculum: 10-100 CFU (Productivity) (ISO/TS 11133-1/2).

Microorganism Growth Remarks

Pseudomonas aeruginosa ATCC 27853 Good Gas (-)

Enterococcus faecalis ATCC 29212 Good Gas (-)

Escherichia coli ATCC 8739 Good Gas (+)

Escherichia coli ATCC 25922 Good Gas (+)

Salmonella typhimurium ATCC 14028 Good Gas (-)

Citrobacter freundii ATCC 43864 Good Gas (+)

Left: Uninoculate tube (Control)

Centre: Escherichia coli  ATCC 25922

Right: Salmonella typhimurium ATCC 14028

“Detail”

Lactose BrothArt. No. 02-105