Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended for selective isolation of Campylobacter species from food and environmental specimens and other materials.
Composition**
| Ingredients | g/L |
|---|---|
| HiVeg™™ peptone No. 3 | 15.000 |
| HiVeg™™ extract No. 2 | 2.500 |
| Yeast extract | 5.000 |
| Sodium chloride | 5.000 |
| Agar | 12.000 |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 19.75 grams in 500 ml purified / distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C and aseptically add 5-7 %v/v sterile lysed horse blood or 10%sterile defibrinated sheep blood and rehydrated contents of 1 vial of Blaser-Wang Selective Supplement (FD006) or Skirrow Selective Supplement (FD008). Mix well and pour into sterile Petri plates.
Principle And Interpretation
Campylobacter species are ubiquitous in the environment inhabiting a wide variety of ecological niches (1). Infection with a Campylobacter species is one of the most common causes of human bacterial gastroenteritis (1). Most species are found in animals (cattle, swine) and cause infertility and abortion (2). C. jejuni was originally isolated on a blood-containing media with antibiotics (3). Skirrow described a selective medium for Campylobacter species consisting of Blood Agar Base No. 2 supplemented with horse blood and antibiotics (4). Subsequently, Blaser et al isolated C.jejuni on Brucella Agar supplemented with sheep blood and four antibiotics (5). Later on, a fifth antibiotic, cephalothin was added to improve the selectivity of the medium by inhibition of accompanying faecal bacteria (6). Campylobacter Agar Base is recommended by APHA for selective isolation of Campylobacter species (7). Campylobacter HiVeg™™ Agar Base is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/ TSE risks. Campylobacter Agar Base is well supplemented to support luxuriant growth of Campylobacter species. Osmotic equilibrium of the medium is maintained by sodium chloride. Blood serves as an additional source of nutrients including X-factor. The antibiotic supplements namely Blaser-Wang (FD006) and Skirrow (FD008) markedly reduce the growth of normal enteric bacteria while enhancing the growth and recovery of C.jejuni from faecal specimens. Amphotericin B in Blaser-Wang supplement greatly or completely inhibits growth of fungi. C.jejuni colonies appear non-haemolytic, flat and gray with an irregular edge or raised and round with a mucoid appearance. Some strains may appear tan or slightly pink. Swarming may be observed on moist surfaces. Incubation at 35-37°C may show a delayed growth of C.jejuni cultures. Incubating the plates at 42°C can fasten this. The contaminated food sample (10 to 25 grams) is enriched in Campylobacter Enrichment Broth Base (M899 + FD042). The broth is incubated with agitation under a micro aerobic atmosphere for 16-18 hrs. The enrichment culture is then plated onto the selective media i.e. Campylobacter Agar Base (MV994) (7).
Type of specimen
Food and dairy samples; Environmental samples.
Specimen Collection and Handling:
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (8,9,10). 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 :
- Due to nutritional variations, some strains may show poor growth.
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
Gelling
Firm, comparable with 1.2% Agar gel.
Colour and Clarity of prepared medium
Basal medium: Yellow coloured clear gel After addition of 5-7% v/v lysed blood: Reddish brown coloured opaque gel forms in Petri plates
Reaction
Reaction of 3.95% w/v aqueous solution at 25°C. pH: 7.4±0.2
pH
7.20-7.60
Cultural Response
Cultural characteristics observed under reduced oxygen atmosphere after an incubation at 35-37°C for 24-48 hours. Blaser-Wang Selective Supplement (FD006) or Skirrow Selective Supplement (FD008)
| Organism | Growth w/ added FD006 | Growth w/ added FD008 |
|---|---|---|
| Candida albicans ATCC 10231 (00054*) | none - poor | moderate |
| Campylobacter jejuni ATCC 29428 (00156*) | good-luxuriant | good-luxuriant |
| Escherichia coli ATCC 25922 (00013*) | none - poor | none - poor |
| Enterococcus faecalis ATCC 29212 (00087*) | none - poor | none - poor |
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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (11,12).
| Product Name | Campylobacter HiVeg™ Agar Base |
|---|---|
| SKU | MV994 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Koneman E. W, Allen S. D., Janda W. M, Schreckenberger P. C., Winn W. C. Jr, 1992, Colour Atlas and Textbook ofClinical Microbiology, 4th Edition, J. B. Lippincott Company. |
| Customized Product Available | No |
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