Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate, Thane(W)-400604, MH, India
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HiEncap™ Buffered Peptone Water
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EC1494ICCL
Recommended as a pre-enrichment medium of Enterobacteriaceae organisms such as Salmonella and Cronobacterium species from food and animal feeding stuffs, water, milk, milk products and other products. The composition and performance criteria of this medium are as per the specifications laid down in ISO 6579-2017, ISO 21528-1:2004, ISO 22964-2017.
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Intended use
Recommended as a pre-enrichment medium of Enterobacteriaceae organisms such as Salmonella and Cronobacterium species from food and animal feeding stuffs, water, milk, milk products and other products. The composition and performance criteria of this medium are as per the specifications laid down in ISO 6579-2017, ISO 21528-1:2004, ISO 22964-2017.
Composition
ISO 6579-1:2017, ISO 6887-1:2017 (Ε), ISO 21528-1:2017, ISO 22964:2017, Specification - Buffered peptone water (BPW)
| Ingredients | Gms / Litre |
|---|---|
| Enzymatic digest of casein | 10.000 |
| Sodium chloride | 5.000 |
| Disodium hydrogen phosphate, dodecahydrate, (Na2HPO4.12H2O) | 9.000 |
| Potassium dihydrogen phosphate (KH2PO4) | 1.500 |
| Final pH (at 25°C) | 7.0±0.2 |
Formula adjusted, standardized to suit performance parameters
# Equivalent to Enzymatic digest of casein
Directions
Each capsule contains 5.02 gms of dehydrated medium. Suspend 1 capsule in 250ml (4 capsules in 1000 ml) distilled or purified water. Heat to boiling to dissolve the medium completely. Dispense as desired and sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
Principle And Interpretation
Microorganisms that are subjected to environmental stresses may become structurally or metabolically damaged or injured. These microorganisms are unable to replicate in selective environments. Therefore these injured organisms must be resuscitated or permitted to repair the damage by incubation in an appropriate, non-selective environment (10). Edel and Kampelmacher (1) noted that sub-lethal injury to Salmonellae may occur in many food preservation processes. Pre-enrichment in Buffered Peptone Water (M14941) at 35°C for 18-24 hours results in repair of injured cells (9). The buffering system prevents bacterial damage due to change in the pH of the medium. ISO committee has also recommended this pre-enrichment medium for the detection of Enterobacteriaceae (3), Salmonella (4) and Cronobacter (6) species from from food stuffs and other materials. It is also recommended as a diluent for enumerations of all microorganisms (7) and Listeria species(8).
Type of specimen
ISO 6579-1:2017/ ISO 6887-1:2017/ ISO 11290-1:2017/ ISO 21528-1:2017/ ISO 22964:2017
Food samples including milk and milk products, in animal feed, in animal faeces, and in environmental samples from the primary production stage.
Specimen Collection and Handling:
Processesing: ISO 6887-1:2017 (7) / ISO 11290-1:2017 (8)
Dilution samples: Preparation of test samples, initial suspension and decimal dilutions for microbiological examination
Processesing: ISO 6579-1:2017 (4)
Pre-enrichment: Samples (25 grams in 225 ml) are preenriched in Buffered Peptone Water (M14941) and incubated at 34°C to 38°C for 18 h ± 2 hours.
Selective enrichment: 0.1 ml of pre-enriched sample is inoculated in 10 ml RVS Broth (M14481) or MSRV Agar (M1428) and incubated at 41.5 ± 1°C for 24 ± 3 hours and 1 ml of culture is inoculated in MKTTn broth (M14961) and incubated at 37±1°C for 24 ± 3 hours.
Isolation: The culture thus obtained is then plated on XLD Agar, Modified (M031I) and incubated at 37° 1°C for 24 ± 3 hours. Simultaneously plating on second isolation agar is carried out.
Confirmation: Biochemical and serological tests are performed for confirmation.
Processesing: ISO 21528-1:2017 (6)
Pre-enrichment: Samples (10 grams in 90 ml) are preenriched in Buffered Peptone Water (M14941) and incubated at 37° 1°C for 18 h ± 2 hours.
Isolation: The culture thus obtained is then plated on Violet red bile glucose (VRBG) agar (M1684) and incubated at 37 ± 1 °C for 24±2hours.
Confirmation: Biochemical and serological tests are performed for confirmation.
Processesing: ISO 22964:2017 (5)
Pre-enrichment: Samples (10 grams in 90 ml) are preenriched in Buffered Peptone Water (M14941) and incubated at 34° C to 38°C for 18 h ± 2 hours.
Selective enrichment: 0.1 ml of pre- enriched sample is inoculated in 10ml Cronobacter Selective Broth (M17861) and incubated at 41.5 ± 1°C for 24±2hours.
Isolation: The culture thus obtained is then plated on HiCrome™ Cronobacter Isolation Agar(CCI Agar)(M20621) and incubated at 41.5 ± 1°C for 24±2hours.
Confirmation: Biochemical and serological tests are performed for confirmation.
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 to individual safety data sheets.
Limitations :
- Individual organisms differ in their growth requirement and may show variable growth patterns in the medium.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
- Further biochemical tests must be carried outfor confirmation.
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: Gelatin capsule containing,cream to yellow granular media
Colour and Clarity of prepared medium: Light yellow coloured clear solution without any precipitate
Reaction: Reaction of 2.0% w/v aqueous solution at 25°C. pH: 7.0±0.2
pH: 6.80-7.20
Cultural Response
ISO 6887-1:2017
Dilution: Recovery of ±30% of the original count (recovered on Tryptone Soya Agar, M290), when the inoculated sample holding time is 45 minutes to 1 hour at 20-25°C. The plates are incubated at at 37°±2°C for 18 h ± 2 hours.
| Organism | Inoculum CFU) | Recovery |
|---|---|---|
| Escherichia coli ATCC 8739 (00012*) | 50-100 | ±30% of the original count |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | ±30% of the original count |
| Staphylococcus aureus ATCC 6538 (00032*) | 50-100 | ±30% of the original count |
| Staphylococcus aureus ATCC 25923 (00034*) | 50-100 | ±30% of the original count |
Dilution: Recovery of ±30% of the original count (recovered on Tryptone Soya Agar, M290), when the inoculated sample holding time is 1 hour ± 5 minutes at 20 ± 2°C. The plates are incubated at at 37°±2°C for 18 h ± 2 hours.
| Organism | Inoculum | Recovery |
|---|---|---|
| Listeria monocytogenes ATCC 13932 (00021*) | 50-100 | ±30% of the original count |
| Listeria monocytogenes ATCC 35152 (00109*) | 50-100 | ±30% of the original count |
ISO 6579-1:2017 & ISO 21528-1:2017
Productivity: Cultural characteristics observed after an incubation at at 34°C to 38°C for 18 h ± 2 hours.
| Organism | Inoculum CFU) | Growth |
|---|---|---|
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | good-luxuriant |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | good-luxuriant |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | good-luxuriant |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | good-luxuriant |
ISO 22964:2017
Productivity: Cultural characteristics observed after an incubation at at 34°C to 38°C for 18 h ± 2 hours.
| Organism | Inoculum | Growth |
|---|---|---|
| Cronobacter sakazakii ATCC 29544 (00214*) | 50-100 | good-luxuriant |
| Cronobacter muytjensii ATCC 51329 (00213*) | 50-100 | 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 15-25°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 (2,3).
| Product Name | HiEncap™ Buffered Peptone Water |
|---|---|
| SKU | EC1494ICCL |
| Application/ Industry | Food |
| Product Type | HiEncap™ |
| Physical Form | Granulated Media in Gelatin Capsule |
| Origin | Animal |
| Packaging type | HDPE Jar |
| References | 1. Vanderzant C. and Splittstoesser D. F., (Eds.), 1992, Compendium of Methods for the Microbiological Examination of Foods, 3rd Ed., APHA, Washington, D.C. 2.Edel W. and Kampelmacher E. H., 1973, Bull. Wld. Hlth. Org., 48: 167. 3.Angelotti R., 1963, “Microbiological Quality of Foods”, Academic Press, New York. 4.Sadovski A. Y., 1977, J. Food Technol., 12.85. 5.International Organization for Standardization (ISO), 2002, Draft ISO/DIS, 6579 |
| Customized Product Available | No |
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