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Sabouraud Dextrose Agar
Intended Use
Recommended for cultivation of C.albicans in accordance with the harmonized method of USP/EP/BP/JP.
Composition
| Ingredients | g/L |
|---|---|
| Dextrose (Glucose) | 40.000 |
| Mixture of Peptone and Tryptone (1:1)# | 10.000 |
| Agar | 15.000 |
| pH after sterilization( at 25°C) | 5.6±0.2 |
Mixture of Peptic digest of animal tissue and Pancreatic digest of casein (1:1)#
**Formula adjusted, standardized to suit performance parameters
Directions
Sabouraud Dextrose Agar is a ready to use solid media in glass bottle. The medium is pre-sterilized, hence it does not need sterilization. Medium in the bottle can be melted either by using a pre-heated water bath or any other method. Slightly loosen the cap before melting. When complete melting of medium is observed dispense the medium in tubes as butts /slants or in plates as desired and allow to solidify. If on plate, either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically.
Principle And Interpretation
Fungi were among the first microorganisms recognized because some of the fruiting structures, such as the mushrooms, are large enough to be seen without a microscope. Fungi can be grouped simply on the basis of morphology as either yeasts or moulds (1). Fungal diseases that occur on the skin, hair and mucous membrane are called superficial mycoses, and the organism that cause them, the dermatophytes (2). Where fungi are to be isolated, it is good practice to use a medium that favors their growth but is not optimal for the growth of bacteria.
Sabouraud Dextrose Agar is Carliers modification (3) of the formulation described by Sabouraud (4) for the cultivation of fungi (yeasts, moulds), and aciduric microorganisms. Sabouraud Dextrose Agar is recommended for microbiological examination of non-sterile products in accordance with the harmonized method of USP/EP/BP/JP (5,6,7,8). This medium is also employed in microbial limit tests in pharmaceutical testing, food, cosmetics.
Peptone and Tryptone provides carbonaceous, nitrogenous compounds, long chain amino acids, vitamins and other essential growth nutrients. Dextrose (Glucose) provides an energy source. High dextrose concentration and low pH favors fungal growth and inhibits contaminating bacteria from clinical specimens (10).
Some pathogenic fungi may produce infective spores, which are easily dispersed in air, so examination should be carried out in safety cabinet. For heavily contaminated samples, the plate must be supplemented with inhibitory agents for inhibiting bacterial growth. Growth of white colonies may be indicative of presence of Candida albicans. The total combined yeast and molds count is considered to be equal to the number of colony forming unit found using this medium, if bacterial colonies are detected they are counted as part of total yeast and mold count. In case the bacterial colonies exceeds the acceptance criterion, then antibiotics can be supplemented in this medium.
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per pharmaceutical guidelines (5,6,7,8). 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 handing specimens and culture. Standard guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
- Some pathogenic fungi may produce infective spores which are easily dispersed in air, so examination should be carried out in safety cabinet
- Further biochemical tests should 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: Sterile glass bottle containing clear to slightly opalscent Sabouraud Dextrose Agar.
Colour of medium: Light yellow to amber coloured medium.
Quantity of medium: 25ml of medium in glass bottle.
pH: 5.40-5.80
Sterility Check: Passes release criteria
Growth Promotion Test
Growth Promotion was carried out in accordance with the harmonized method of ICH (USP/EP/BP/JP), after an incubation at 30-35 °C for 24-48 hours. Recovery rate is considered as 100% for bacteria growth on Soybean Casein Digest Agar and fungus growth on Sabouraud Dextrose Agar
Growth Promoting Properties
Growth of microorganism comparable to that previously obtained with previously tested and approved lot of medium occurs at the specified temperature for not more than the shortest period of time specified inoculating <= 100 cfu (at 30-35°C for <=24 hours).
Indicative properties
Colonies are comparable in appearance and indication reaction to those previously obtained with previously tested and approved lot of medium occurs for the specified temperature for a period of time within the range specified inoculating <=100cfu (at 30-35°C for 24-48 hours).
Cultural Response
| Organism | Inoculum (CFU) | Growth | Observed Lot Recovery value (CFU) | Incubation temperature | Incubation period | |
|---|---|---|---|---|---|---|
| Growth Promotion + Indicative | ||||||
| Candida albicans ATCC 10231 (00054*) | 50-100 | Luxuriant (white colonies) | 35-100 | >=70% | 30-35 °C | 24-48 hrs |
| Growth Promotion + Total yeast and mould count | ||||||
| Candida albicans ATCC 10231 (00054*) | 50-100 | luxuriant | 35-100 | >=70% | 20-25 °C | <=5 d |
| Aspergillus brasiliensis ATCC 16404 (00053*) | 50-100 | luxuriant | 35-100 | >=70% | 20-25 °C | <=5 d |
Key: (#) - Formerly known as Aspergillus niger, (*) - corresponding WDCM numbers
Storage and Shelf life
On receipt store between 15-30°C. Use before expiry date on the label. 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 (10,11).
| Product Name | Sabouraud Dextrose Agar |
|---|---|
| SKU | SMH063XXV |
| Customized Product Available | No |

