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Dehydrated culture media

WL Nutrient Broth

For the determination of microbial flora in beer fermentation processes and manufacturing.

Cat. No.
LC-1572
Format
500 g
Storage
2 to 25 °C
Minimum guaranteed shelf life
15 months

WL Nutrient Broth, based on the Green and Gray formulation, is recommended for the control of industrial fermentations, particularly the manufacturing of beer. With a pH of 5.5, true counts of beer yeasts can be made. With a pH of 6.5, the medium is ideal for bakery and distilled spirit yeasts.

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Cat. No.FormatPrice
LC-1572500 g

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Key features

Applications

Selective enumeration of microorganisms of the brewing industry.

Industry

Alcoholic beverages, fermentation, food.

Preparation

60.25 g in one liter of distilled water; boil for one minute, sterilize in autoclave at 121 °C for 15 minutes.

Principles and uses

Tryptone provides nitrogen, vitamins, minerals and amino acids essential for growth. Dextrose is the fermentable carbohydrate providing carbon and energy. Yeast extract is a source of vitamins, particularly of the B-group. Monopotassium phosphate is the buffer. Potassium, calcium and ferric chlorides all provide essential ions for the osmotic balance. Manganese and magnesium sulfates are a source of divalent cations. Bromocresol green is the pH indicator.

Technical specifications

SolubilityWithout residues
AppearanceFine powder
Color of the dehydrated mediumClear toasted with a blue tint
Color of the prepared mediumLight blue
Final pH (25 °C)5.5 ± 0.2
Amount per litre60.25 g
Storage2 to 25 °C

Formula

Ingredientg/L
Dextrose50
Bromocresol green0.022
Calcium chloride0.125
Ferric chloride0.0025
Magnesium sulfate0.125
Manganese sulfate0.0025
Monopotassium phosphate0.55
Potassium chloride0.425
Tryptone5
Yeast extract4

Preparation and use

Preparation

Suspend 60.25 grams of the medium in one liter of distilled water. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Sterilize in autoclave at 121 °C for 15 minutes. Cool to 45-50 °C, mix well and dispense into tubes.

Instructions for use

Inoculate and incubate at a temperature of 30 °C and observe after 24-48 hours.

The formulae of WL Nutrient (WLN) and WL Differential (WLD) are used in conjunction, as with WLN Agar the bacteria may not be detected unless the number of yeast cells is very small. 1 plate of WLN and 2 plates of WLD must be used.

  • The WLN Agar plate is incubated aerobically for the total plate count of yeasts.
  • One of the WLD Agar plates is incubated aerobically for acetic acid bacteria: Flavobacterium, Proteus, thermophilic bacteria and others.
  • The second WLD plate is incubated anaerobically for the investigation of lactic acid bacteria and species of Pediococcus.

All plates are incubated, in general, at 25 °C in the case of beer, and at 30 °C for bakery and malt alcoholic yeasts. Plates are incubated for 2-10 days and up to 2 weeks, according to the flora present. Counts are made at regular intervals during this period.

Quality control

Microbiological test

Incubation conditions: 30 °C / 24-48 h.

MicroorganismSpecification
Escherichia coliATCC 25922Moderate growth
Lactobacillus fermentumATCC 9338Moderate growth
Saccharomyces cerevisiaeATCC 9763Good growth

Bibliography

  • Green, S.R. and P.P. Gray 1950. Paper read at American Society of Brewing Chemists Meeting. Wallerstein Lab. Commun. 12:43.
  • Green, S.R. and P.P. Gray 1950. A differential procedure applicable to bacteriological investigation in brewing. Wallerstein Lab. Commun. 13:357.
  • MacFaddin J.D. 1985. Media for isolation-cultivation-identification-maintenance of medical bacteria, vol. 1. p. 854-856. Williams & Wilkins, Baltimore, MD.

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Shipped under controlled temperature when the product requires it.

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