Buffered HiCynth™ Peptone Water w/ NaCl

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MCD1275
Recommended as a diluent for carrying out microbial limit test from clinical and non-clinical specimens.


Intended use

Recommended as a diluent for carrying microbial limit test from various specimens.

Composition**

Ingredients Gms/Litre
HiCynth™ Peptone No.1* 1.000
Potassium dihydrogen phosphate 3.560
Disodium hydrogen phosphate 7.230
Sodium chloride 4.300
Final pH (at 25°C) 7.0±0.2

**Formula adjusted, standardized to suit performance parameters

* Chemically defined peptone

Directions

Suspend 16.09 grams in 1000 ml purified / distilled water. Heat if necessary to dissolve the medium completely. Add 0.1 to 1% w/v polysorbate 20 or 80 if desired (depending on the type of sample to be diluted). Dispense in tube or flasks as desired. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.

Principle And Interpretation

Buffered Peptone Water is a pre-enrichment medium designed to help recovery of sub-lethally damaged Salmonellae before transfer to a selective medium. This pre-enrichment medium is free from inhibitors and is well buffered and provides conditions for resuscitation of the cells that have been injured by processes of food preservation. It was noted by Edel and Kampelmacher (1) that sub-lethal injury to Salmonella may occur due to food preservation techniques involving heat, desiccation, high osmotic pressure, preservatives or pH changes. Buffered Peptone Water during the pre-enrichment period helps in recovery of injured cells that may be sensitive to low pH (2). This is particularly important for vegetable specimens, which have low buffering capacity. These media can be used for testing dry poultry feed (3). In a survey involving isolation of Salmonellae from meat that had been artificially contaminated with sub-lethally injured organisms. Pre-enrichment in Buffered Peptone Water at 37°C for 18 hours before selection in Tetrathionate Brilliant Green Bile Broth (M1255) showed superior results compared with direct selection method. Lactose Broth is frequently used as a pre-enrichment medium but it may be detrimental to recovery of Salmonellae (4). The composition of Buffered Peptone Water with NaCl medium is as per IP and EP specifications recommended to dilute the sample for microbial examination (5,6). Depending on the amount of fat in the sample to examine the kind and quantity of emulsifying agent to be used.

Buffered HiCynth™ Peptone Water is prepared by replacing animal and vegetable peptones with chemically defined peptones to avoid BSE/TSE risks associated with animal peptones. These pre-enrichment media contain HiCynth™ Peptone No.1 as a source of carbon, nitrogen, vitamins and minerals. Sodium chloride maintains the osmotic balance and phosphates buffer the medium. The broth is rich in nutrients and produces high resuscitation rates for sublethally injured bacteria and supports intense growth. The phosphate buffer system prevents bacterial damage due to changes in the pH of the medium. Inoculate 10 grams specimen in 50 ml of these media and incubate at 35-37°C for 18 hours. Transfer 10 ml from this medium to 100 ml of Tetrathionate Broth (M032) and incubate at 43°C for 24-48 hours and then subculture on selective plating media. Examine the plates for characteristic Salmonella colonies.

Type of specimen

Food and dairy samples

Specimen Collection and Handling:

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (3,7,8,9). After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions :

For professional use only. 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.

Limitations :

  1. This medium contains low nutrients and hence is not recommended for the growth of organisms.
  2. Further recovery on solid media and biochemical or serological testing of pure colony is required for complete identification.

Performance and Evaluation

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

Quality Control

Appearance
White to cream homogeneous free flowing powder

Colour and Clarity of prepared medium
Colourless to pale yellow clear solution without any precipitate

pH
6.80-7.20

Cultural response
Cultural characteristics observed after recovery on Soybean Casein Digest Agar after an incubation at 30-35°C for 18-24 hours for bacteria and Sabouraud Dextrose Agar at 30-35°C for 24-48 hours.

Organism Inoculum
(CFU)
Recovery
within 2 hours
of incubation
Recovery
within 4 hours
of incubation
Recovery
within 24
hours of
incubation
Escherichia coli
ATCC 8739 (00012*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Escherichia coli
ATCC 25922 (00013*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Escherichia coli NCTC
9002
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Staphylococcus aureus
subsp. aureus ATCC
6538 (00032*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Staphylococcus aureus
subsp. aureus ATCC
25923 (00034*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Pseudomonas
aeruginosa
ATCC 9027
(00026*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Pseudomonas
aeruginosa
ATCC 27853
(00025*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Salmonella Typhimurium
ATCC 14028 (00031*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Salmonella Abony
NCTC 6017 (00029*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Bacillus subtilis subsp.
spizizenni ATCC 6633
(00003*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Micrococcus luteus
ATCC 9341
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Candida albicans
ATCC 10231 (00054*)
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at
2-8°C)
Candida albicans
ATCC 2091
50-100 no decrease in
colony count
no decrease in
colony count
no decrease in
colony count
(stored at 2-8°C)

Key: (*) - Corresponding WDCM Numbers

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 15-30°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order 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 (10,11).

Reference

  1. Edel and Kampelmacher, 1973, Bull. W.H.O., 48:167.
  2. Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination of Foods, 5th Ed., American Public Health Association, Washington, D.C.
  3. Sadovski, 1977, J. Food Technol., 12:85.
  4. Angelotti, 1963, Academic Press, New York, N.Y.
  5. European Pharmacopoeia, 2022, 10 th volume, European Directorate for the quality of medicines & Healthcare.
  6. Indian Pharmacopoeia, 2022, Indian Pharmacopoeia Commission, Ministry of Health and Family Welfare Government of India.
  7. American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., Washington D.C.
  8. Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed., APHA Inc., Washington, D.C.
  9. Juven, Cox, Bailey, Thomson, Charles and Schutze, 1984, J. Food Prot., 47:299.
  10. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
  11. Jorgensen, J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1.
More Information
Product Name Buffered HiCynth™ Peptone Water w/ NaCl
SKU MCD1275
Product Type HiCynth™
Physical Form Powder
Origin Chemically defined (HiCynth™)
Packaging type HDPE
References 1. Edel and Kampelmacher, 1973, Bull. W.H.O., 48:167.2.Sadovski, 1977, J. Food Technol., 12:85.3.Juven, Cox, Bailey, Thomson, Charles and Schutze, 1984, J. Food Prot., 47:299.4.Angelotti, 1963, Academic Press, New York, N.Y.5.Indian Pharmacopoeia, 1997, Ministry of Health and Family Welfare,Govt. of India, Vol. 2.6.European Pharmacopoeia, 2008, European Directorate For The Quality of Medicine7.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., WashingtonD.C.8.Downes F. P. and Ito K., (Ed.), 2001, Compendium of Methods for the Microbiological Examination of Foods, 4th Ed.,American Public Health Association, Washington, D.C.9.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C.10.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition.11. Jorgensen, J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1.
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