Modified Scholten's Broth

SKU:
M2165I
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for detection and enumeration of bacteriophages from water samples. The composition and performance criteria are as per specifications laid down in ISO 10705-2:2000.

g/L : 29.03 17.22 Lit/500G



Intended Use

Recommended for detection and enumeration of bacteriophages from water samples. The composition and performance criteria are as per specifications laid down in ISO 10705-2:2000.

Composition

As per ISO 10705-2:2000 Modified Scholten's Broth M2165I
Ingredients g / L Ingredients g / L
Peptone 10.000 Peptone 10.000
Yeast extract 3.000 Yeast extract 3.000
Meat Extract 12.000 HM Extract# 12.000
Sodium chloride 3.000 Sodium chloride 3.000
Sodium carbonate (15%) 5.00 ml Sodium carbonate 0.750
Magnesium chloride (100 g in 50 ml water) 0.30 ml Magnesium chloride, hexahydrate 0.600
pH after sterilization 7.2±0.5 pH after sterilization 7.2±0.5

**Formula adjusted, standardized to suit performance parameters.

# Equivalent to Meat extract.

Directions

Suspend 29.03 grams (the equivalent weight of dehydrated medium per litre) in 1000 ml of purified/distilled water. Heat if necessary to dissolve the medium completely. Dispense into tubes or flasks or as desired. Sterilize by autoclaving at 121±3°C for 15 minutes.

Principle and Interpretation

The formulation of Modified Scholten's Broth is as described in ISO 10705-2 (1). Bacteriophages, also known as phages, are viruses that infect and replicate only in bacterial cells. Like all viruses, bacteriophages are very species-specific with regard to their hosts and usually only infect a single bacterial species or even specific strains within a species. Phages are clinically significant for several reasons.

Peptone, HM Extract and yeast extract in the medium provides the necessary nitrogen compounds, carbon, vitamins and also some trace ingredients necessary for the growth of bacteria. Sodium chloride maintains the osmotic equilibrium of the medium. Magnesium chloride present in the medium raises the osmotic pressure. Sodium carbonate and magnesium chloride help to maintain the pH of the medium.

Type of Specimen

Water samples.

Specimen Collection and Handling

ISO 10705-2:2000 (1)

Rehydrate the content of a lyophilized ampoule of the reference culture of the host strain in 3 ml of Modified Scholten's Broth (M2165I). Transfer the suspension to 50 ml of Modified Scholten's Broth (M2165I) and incubate at 36±2°C for (20±4) h.

Add 1 ml of inoculum culture to each culture tube containing the aliquots of sample and Scholten's Agar (M2134I). Incubate the plates at 36±2°C for (18±2) h. Count the number of plaques on each plate within 4 h after finishing incubation, using indirect oblique light.

Warning and Precautions

Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/face protection. Follow good microbiological lab practices while handling specimens and culture.

Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.

Performance and Evaluation

Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.

Quality Control

Appearance: Cream to yellow homogeneous free flowing powder.

Colour and Clarity of prepared medium: Light amber colored clear to slightly opalescent solution.

Reaction: Reaction of 2.90% w/v aqueous solution at 25°C. pH after sterilization: 7.2±0.5.

pH after sterilization: 6.70-7.70.

Cultural Response: Cultural characteristics observed after an incubation at 36±2°C for (20±4) hours.

Technical Specifications

Inoculum Escherichia coli ATCC 25922 (00013*)
Inoculum (CFU) 50-100
Growth good-Luxuriant
Key : (*) Corresponding WDCM numbers.

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle inorder to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.

Disposal

User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques

More Information
Product Name Modified Scholten's Broth
SKU M2165I
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