Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Campylobacter Enrichment HiVeg™ Broth Base (Preston Enrichment HiVeg™ Broth Base)
Recommended for selective enrichment and cultivation of Campylobacter species.
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Intended Use:
Recommended for selective enrichment and cultivation of Campylobacter species from various samples.
Composition
| Ingredients | g/L |
|---|---|
| HiVeg™™ peptone | 10.000 |
| HiVeg™ extract | 10.000 |
| Sodium chloride | 5.000 |
| Final pH (at 25°C) | 7.5±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 12.5 grams in 470 ml purified / distilled water. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to room temperature and aseptically add sterile 25 ml lysed horse blood and reconstituted contents of 1 vial of PRTC Selective Supplement(FD042). Cool to 45-50°C. Mix well and dispense as desired.
Principle And Interpretation
Balton and Robertson (1) described this as a selective medium for the cultivation of Campylobacter species. It is recommended by APHA (2) for enrichment of thermotolerant Campylobacter species from foods. Preston Enrichment Broth has a rich basal medium to aid resuscitation of sublethally damaged Campylobacter. Preliminary incubation of the medium complete with antibiotics for 4 hours at 37°C was recommended to aid resuscitation of injured organisms followed by 42°C for 18-48 hours (3). HiVeg™ peptone and HiVeg™ extract in the medium provide nitrogen, vitamins and minerals necessary to support bacterial growth. Sodium chloride provides essential ions. Campylobacter Enrichment HiVeg™ Broth Base is prepared by completely replacing animal based peptone with vegetable peptones to avoid BSE/TSE risks associate with animal peptones. Preston Selective Supplement (FD042) contains antibacterial and antifungal agents. Polymyxin B is active only against gram-negative bacteria and Proteus species are sometimes resistant. Trimethoprim usually inhibits Proteus species as well as other gram-negative bacteria. Rifampicin is also active against gram-negative organisms. Cycloheximide acts as antifungal agent. Direct plating without enrichment is adequate for fresh faecal samples, fecal contents or intestinal specimens as high numbers of the organisms may be anticipated. For food samples enrichment is required. Humphrey (1989) suggested that pre-enrichment at 37°C should be continued for 4 hours and that addition of all antibiotics should be delayed until the 4 hours pre- enrichment had been completed. Enrichment medium with rifampicin was recommended in parallel with similar plating medium. The Campylobacter species grow well in microaerobic conditions i.e. in 5% O2 at 42°C in about 48 hours. Addition of about 4 drops of glycerol to a filter paper kept within the jar/container will hamper confluent and swarming growth of Campylobacter (1).
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 (2,4). After use, contaminated materials must be sterilized by autoclaving before discarding.
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.
Limitations :
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- Further isolation and biochemical tests must be carried out for 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: Cream to yellow homogeneous free flowing powder
Colour and Clarity: Basal medium : Light yellow coloured clear solution. After addition of sterile lysed horse blood: Cherry red coloured opaque solution in tubes
Reaction: Reaction of 2.5% w/v aqueous solution at 25°C. pH: 7.5±0.2
pH 7.30-7.70
Cultural Response: Cultural characteristics observed with added 25ml sterile lysed horse blood and PRTC Selective Supplement (Preston Selective Supplement), (FD042), after an incubation at 42°C for 48 hours (5% O2+10% CO2 + 85% N2).
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| Bacillus cereus ATCC 10876 | >=104 | inhibited |
| Campylobacter coli ATCC 33559 (00072*) | 50-100 | good-luxuriant |
| Campylobacter jejuni ATCC 29428 (00156*) | 50-100 | good-luxuriant |
| Campylobacter lari ATCC 35221 | 50-100 | good-luxuriant |
| Escherichia coli ATCC 25922 (00013*) | >=104 | inhibited |
| Proteus mirabilis ATCC 25933 | >=104 | inhibited |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=104 | inhibited |
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 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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (5,6).
| Product Name | Campylobacter Enrichment HiVeg™ Broth Base (Preston Enrichment HiVeg™ Broth Base) |
|---|---|
| SKU | MV899 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Balton F.J. and Robertson L., 1982, J. Clin. Pathol., 35:462. |
| Customized Product Available | No |
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