Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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B.T.B. Lactose HiCynth™ Agar, Modified (Lactose Blue HiCynth™ Agar)
Recommended for diffrentiation of lactose fermenting and non fermenting bacteria belonging to Enterobacteriaceae.
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Intended Use
Recommended for differentiation of lactose fermenting and non-fermenting bacteria belonging to Enterobacteriaceae.
Composition**
| Ingredients | g/L |
|---|---|
| HiCynth™ Peptone No.1* | 3.500 |
| HiCynth™ Peptone No.5* | 3.500 |
| Sodium chloride | 5.000 |
| Lactose | 15.500 |
| Bromo thymol blue | 0.040 |
| Agar | 13.000 |
| Final pH (at 25°C) | 7.0±0.2 |
**Formula adjusted, standardized to suit performance parameters
* -Chemically Defined peptones
Directions
Suspend 40.54 gram 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. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Reactions with lactose are of great practical importance for the primary isolation of Enterobacteria from specimens. The specimens e.g. faeces is usually plated on a lactose-containing medium on which lactose fermenters and lactose non fermenters form coloured and pale colonies respectively due to the dye incorporated. This procedure makes an immediate presumptive distinction between colonies of the true intestinal pathogens possible. Salmonella and Shigella, do not ferment lactose while the common intestinal commensals, Escherichia and Klebsiella, which do ferment lactose (1). Lactose Blue Agar is used for differentiating lactose fermenting and non-fermenting bacteria belonging to the family Enterobacteriaceae.
This medium is prepared by completely replacing animal peptones with chemically defined peptones to avoid BSE/TSE / GMO risks associated with animal peptones. HiCynth™ Peptone No.1 and HiCynth™ Peptone No.5 provide essential nutrients for bacterial metabolism. Lactose provides a fermentable carbohydrate source for the enteric bacteria. Bromo thymol blue is the pH indicator for indicating acid production due to carbohydrate fermentation. The dye turns yellow at acidic pH and imparts yellow colour to the colony. Alkalinization produces a blue coloration. Winkle (2) recommended addition of 0.28g/1 metachrome yellow to suppress the swarming of Proteus species.
Type of specimen
Isolated Microorganisms
Specimen Collection and Handling:
For microbial specimens, follow appropriate techniques for handling specimens as per established guidelines. 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 biochemical and serological tests must be carried out for complete identification.
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 greenish yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.3% Agar gel.
Colour and Clarity of prepared medium
Green coloured, clear to slightly opalescent gel forms in Petri plates.
Reaction
Reaction of 4.05% w/v aqueous solution at 25°C. pH: 7.0±0.2
pH
6.80-7.20
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | >=70% | yellow, opaque |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | >=70% | bluish |
| Salmonella Typhi ATCC 6539 | 50-100 | luxuriant | >=70% | bluish |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | good-luxuriant | >=70% | deep yellow |
Key: (*) Corresponding WDCM numbers.
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 (3,4).
Reference
- Cruikshank R., Duguid J. P., Marmion B. P., Swain R. H. A., (Eds.), 1975, Medical Microbiology, The Practice of Medical Microbiology, 12th Edition, Vol. II, Churchill Livingstone
- Winkle S., 1947, Zbl. Bakt. I. Orig., 152:103.
- Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
- 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.
| Product Name | B.T.B. Lactose HiCynth™ Agar, Modified (Lactose Blue HiCynth™ Agar) |
|---|---|
| SKU | MCD1081 |
| Product Type | HiCynth™ |
| Physical Form | Powder |
| Origin | Chemically defined (HiCynth™) |
| Packaging type | HDPE |
| References | 1. Cruikshank R., Duguid J. P., Marmion B. P., Swain R. H. A., (Eds.), 1975, Medical Microbiology, The Practice of MedicalMicrobiology, 12th Edition, Vol. II, Churchill Livingstone |
| Customized Product Available | No |
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