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HiCrome™ Bacillus HiCynth™ Agar Base
Recommended for isolation and differentiation between various species of Bacillus from a mixed culture in food, clinical and non-clinical samples by chromogenic method.
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Intended use
Recommended for isolation and differentiation between various species of Bacillus from a mixed culture by chromogenic method.
Composition**
| Ingredients | Gms/Litre |
|---|---|
| HiCynth™ Peptone No.3* | 11.000 |
| D-Mannitol | 10.000 |
| Sodium chloride | 10.000 |
| Chromogenic mixture | 3.200 |
| Phenol red | 0.025 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.1±0.2 |
**Formula adjusted, standardized to suit performance parameters
*Chemically defined peptone
Directions
Suspend 49.22 grams in 1000 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. For selective isolation of B.cereus and B.thuringiensis aseptically add rehydrated contents of one vial of PB Selective Supplement (FD324). Mix well and pour into sterile Petri plates.
Principle And Interpretation
Majority of Bacillus species apparently have little or no pathogenic potential and are rarely associated with disease in humans or lower animals. The principal exception to this are Bacillus anthracis, the agent of anthrax, and Bacillus cereus, but a number of other species, particularly those of the B.subtilis group, have been implicated in food poisoning and other human and animal infections (1). Bacillus cereus causes food poisoning due to consumption of contaminated rice (2,3,4), other starchy foods such as potato, pasta and cheese have also been implicated, eye infections and a wide range of other conditions like abscess formation, meningitis, septicemia and wound infection.
HiCrome™ Bacillus Agar is based on the formulation of MYP Agar formulated by Mossel et al (5) used for enumeration of Bacillus cereus and Bacillus thuringiensis when present in large number in certain foodstuffs. HiCrome™ Bacillus HiCynth™ Agar is the modification of the same, prepared by completely replacing animal based peptones with vegetable peptones to avoid BSE/TSE risks associated with animal peptones.
The medium contains HiCynth™ Peptone No.3 which provide nitrogenous compounds. Mannitol serves as the fermentable carbohydrate, fermentation of which can be detected by phenol red. Mannitol fermenting organisms like B.megaterium yield yellow coloured colonies. The chromogenic mixture present in the medium is cleaved by the enzyme β-glucosidase found in B.cereus resulting in the formation of blue colonies. B.thuringiensis also grows as blue/green colonies on this medium as B.cereus and B.thuringiensis are biochemically identical, however B.cereus shows flat colonies with distinct blue centers, while B.thuringiensis shows irregular margins.
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 (6,7,8). 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.
- Slight variation in colour may be observed depending upon the utilization of the substrate by the organism.
- Further serological or biochemical testing can be performed for complete identification.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within expiry period when stored at therecommended temperature.
Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Red coloured, clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 4.92% w/v aqueous solution at 25°C. pH: 7.1±0.2
pH
6.90-7.30
Cultural Response
Cultural characteristics observed after an incubation at 30°C for 24-48 hours.
| Organism | Inoculum (CFU) | Growth w/o addition of FD003 | Recovery w/o addition of FD003 | Growth w/ addition of FD003 | Recovery w/ addition of FD003 | Colour of Colony |
|---|---|---|---|---|---|---|
| Bacillus subtilis subsp. spizizenii ATCC 6633 (00003*) | 50-100 | fair | 20-30% | inhibited | 0% | yellowish green to green colonies |
| Bacillus cereus ATCC 10876 | 50-100 | good-luxuriant | >=50% | good-luxuriant | >=50% | light blue, large, flat colonies with blue centre |
| Bacillus thuringiensis ATCC 10792 | 50-100 | good-luxuriant | >=50% | good-luxuriant | >=50% | light blue, large, flat colonies with irregular margins |
| Bacillus megaterium ATCC 14581 | 50-100 | good-luxuriant | >=50% | inhibited | 0% | yellow, mucoid colonies |
| Bacillus coagulans ATCC 7050 (00002*) | 50-100 | good-luxuriant | >=50% | inhibited | 0% | pink, small, raised colonies |
| Bacillus pumilis ATCC 14884 | 50-100 | good-luxuriant | >=50% | poor | 10-20% | light green to green colonies |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | luxuriant | >=50% | inhibited | 0% | yellow colonies |
| Enterococcus faecalis ATCC 29212 (00087*) | 50-100 | luxuriant | >=50% | inhibited | 0% | light green to green colonies |
Key: (*) - Corresponding WDCM numbers
Storage and Shelf Life
Store between 15-25°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 (9,10).
| Product Name | HiCrome™ Bacillus HiCynth™ Agar Base |
|---|---|
| SKU | MCD1651 |
| Product Type | HiCynth™, HiCrome™ |
| Physical Form | Powder |
| Origin | Chemically defined, HiCynth™ |
| Packaging type | HDPE |
| References | 1.Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Eds.), 2003, Manual of Clinical Microbiology,8th Ed. American Society for Microbiology, Washington, D.C.2.Mossel D. A. A., Koopman M. J. and Jongerium E., 1967, Appl. Microbiol., 15:650.3.Mortimer P. R. and McCann G., 1974, Lancet, 1043.4.Bouza E., Grant S., Jordan C. et al, 1979, Arch. Ophthamol., 97:488.5.Wohlgemuth K., Kirkbride C. A., Bicknell E. J. and Ellis R. P., 1972 Am. Vet. Met, Ass. 161:1691.6.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition. |
| Customized Product Available | No |
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