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Decarboxylase HiVeg® Broth Base, Moeller (Moeller Decarboxylase HiVeg® Broth Base)
To differentiate bacteria on the basis of their ability to decarboxylate the amino acids.
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Intended Use
Recommended with the addition of appropriate L-amino acid for use, to differentiate bacteria on the basis of their ability to decarboxylate the amino acid.
Composition
| Ingredients | g/L |
|---|---|
| HiVeg® peptone | 5.000 |
| HiVeg® extract | 5.000 |
| Dextrose (Glucose) | 0.500 |
| Bromocresol purple | 0.010 |
| Cresol red | 0.005 |
| Pyridoxal | 0.005 |
| Final pH (at 25°C) | 6.0±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Suspend 10.52 grams in 1000 ml purified/distilled water. Add 10 grams of L-Lysine, L-Arginine, L-Ornithine or other L-amino acids as required. When using DL-amino acids, use 2% concentration. When L-Ornithine is added readjustment of pH is required. Heat if necessary to dissolve the medium completely. Dispense in 5 ml amounts in screw-capped tubes and sterilize by autoclaving at 15 lbs pressure (121°C) for 10 minutes.
Principle And Interpretation
Moeller Decarboxylase Broth Base is used for differentiating gram-negative enteric bacilli on the basis of their ability to decarboxylate amino acids. Moeller introduced the Decarboxylase Broth for detecting the production of lysine and ornithine decarboxylase and arginine dihydrolase (1). Prior to Moellers work, bacterial amino acid decarboxylases were studied by Gale (2) and Gale and Epps (3). Production of ornithine decarboxylase is a helpful criterion in differentiating Klebsiella and Enterobacter species. Klebsiella are nonmotile and do not produce ornithine decarboxylase while Enterobacter are motile and produce ornithine decarboxylase except Enterobacter agglomerans (4). Decarboxylase HiVeg® Broth Base, Moeller is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks. This medium contains HiVeg® peptone and HiVeg® extract which provides nitrogenous and cabonaceous compounds, long chain amino acids and other essential nutrients for the growth of bacteria. Dextrose is the fermentable carbohydrate and pyridoxal is the co-factor for the decarboxylase enzyme. Bromo cresol purple and cresol red are the pH indicators in this medium. When the medium is inoculated with the dextrose fermenting bacteria, the pH is lowered due to acid production, which changes the colour of the indicator from purple to yellow. Acid produced stimulates decarboxylase enzyme. Decarboxylation of lysine yields cadaverine while putrescine is produced due to ornithine decarboxylation. Arginine is first hydrolyzed to ornithine which is then decarboxylated to form putrescine. Formation of these amines increases the pH of the medium, changing the colour of the indicator from yellow to purple. If the organisms do not produce the appropriate enzyme, the medium remains acidic, yellow in colour. Each isolate to be tested should also be inoculated into Moeller Decarboxylase Broth Base medium tube lacking the amino acid.
Inoculated tubes must be protected from air with a layer of sterile mineral oil. Exposure to air may cause alkalinization at the surface of the medium which makes the test invalid.
Type of specimen
Food and dairy samples; Water samples
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (5,6,7).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(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 clinical specimens. Safety guidelines may be referred in individual safety data sheets
Limitations
- Some fastidious organisms may show delayed reaction.
- Overlaying with mineral oil is essential for appropraite results.
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: Light yellow to greenish yellow homogeneous free flowing powder
Colour and Clarity of prepared medium: Purple coloured, clear solution without any precipitate in tubes
Reaction: Reaction of 1.05% w/v aqueous solution at 25°C. pH: 6.0±0.2
pH: 5.80-6.20
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for upto 4 days with addition of appropriate amino acids and overlaying with sterile mineral oil.
| Organism | Inoculum (CFU) | Arginine decarboxylation | Ornithine decarboxylation | Lysine decarboxylation |
|---|---|---|---|---|
| Citrobacter freundii ATCC 8090 | 50-100 | variable reaction | variable reaction | negative reaction, yellow colour |
| # Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | negative reaction, yellow colour | positive reaction, purple colour | positive reaction, purple colour |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | variable reaction | variable reaction | positive reaction, purple colour |
| Klebsiella pneumoniae ATCC 13883 (00097*) | 50-100 | negative reaction, yellow colour | negative reaction, yellow colour | positive reaction, purple colour |
| Proteus mirabilis ATCC 25933 | 50-100 | negative reaction, yellow colour | positive reaction, purple colour | negative reaction, yellow colour |
| Proteus vulgaris ATCC 13315 | 50-100 | negative reaction, yellow colour | negative reaction, yellow colour | negative reaction, yellow colour |
| Salmonella Paratyphi A ATCC 9150 | 50-100 | delayed positive reaction/positive reaction,purple colour | positive reaction, purple colour | negative reaction, yellow colour |
| Salmonella Typhi ATCC 6539 | 50-100 | delayed positive reaction/negative reaction | negative reaction, yellow colour | positive reaction, purple colour |
| Serratia marcescens ATCC 8100 | 50-100 | negative reaction, yellow colour | positive reaction, purple colour | positive reaction, purple colour |
| Shigella dysenteriae ATCC 13313 | 50-100 | negative reaction/delayed positive reaction | negative reaction, yellow colour | negative reaction, yellow colour |
| Shigella flexneri ATCC 12022 (00126*) | 50-100 | negative reaction/delayed positive reaction | negative reaction, yellow colour | negative reaction, yellow colour |
| Shigella sonnei ATCC 25931 | 50-100 | variable reaction | positive reaction, purple colour | negative reaction, yellow colour |
Key: (*) Corresponding WDCM numbers. (#) Formerly known as Enterobacter aerogenes
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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (9,10).
| Product Name | Decarboxylase HiVeg® Broth Base, Moeller (Moeller Decarboxylase HiVeg® Broth Base) |
|---|---|
| SKU | MV393 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Moeller V., 1955, Acta Pathol. Microbiol. Scand. 36:158. 2.Gale G. F., 1940, Biochem. J., 34:392. 3.Gale and Epps, 1943, Nature, 152:327. 4.MacFaddin J., 1980, Biochemical Tests for Identification of Medical Bacteria, 2nd ed., Williams and Wilkins, Baltimore. 5.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed.,Washington D.C. 6.Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Inc.,Washington, D.C. 7.Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition. |
| Customized Product Available | No |
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