Asparagine Proline Broth

Asparagine Proline Broth
Asparagine Proline Broth for cultivation of Pseudomonas aeruginosa from water samples by membrane filter technique.

Directions
Suspend 14.5 grams or 23.2 grams in 1000 ml distilled water containing 25 ml or 40 ml ethanol respectively. Heat to boiling to dissolve the medium completely. Distribute as desired in screw-capped bottles. Close the caps so that the seal in the lid just touches the lip of the bottle. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Tighten the caps of the bottles immediately after removal from the autoclave to prevent loss of ethanol by evaporation. It is not advisable to use polypropylene caps without seals. Alternatively, ethanol may be sterilized separately by filtration and then added aseptically to the sterile cooled medium.

Principle And Interpretation
Asparagine Proline Broth contains both the enantiomeric forms of Asparagine, which is readily utilized by Pseudomonas for their growth. Phosphate and sulphates provide the ions for the growth as well as buffers the medium to promote the growth of the organism. When 1 ml of sample is to be analyzed, add 1 ml of sample to 4 ml of single strength medium. If larger portions of the sample (10 ml, 50 ml) are to be used, add the sample to an equal volume of the concentrated medium (23.2 g/l). Incubate at 37 ±1°C for 48 hours. Examine for growth and fluorescence. The growth is further sub cultured on Milk Agar w/ Cetrimide

Quality Control
Reaction: Reaction of 1.45% w/v aqueous solution at 25°C. pH : 7.2±0.2
Appearance: White to cream homogeneous free flowing powder
pH: 7.20-7.40
Colour and Clarity of prepared medium: Colourless clear solution, without any precipitate.
Storage: Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C.

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