HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended Use:
Recommended for sterility testing of turbid or viscous biological products and for cultivation of anaerobes and microaerophiles from various samples.
Composition**
| Ingredients | g/L |
|---|---|
| HiCynth™ Peptone No.3* | 15.000 |
| HiCynth™ Peptone No.5* | 5.000 |
| Dextrose (Glucose) | 5.500 |
| Sodium chloride | 2.500 |
| L-cystine | 0.500 |
| Sodium thioglycollate | 0.500 |
| Final pH (at 25°C) | 7.1±0.2 |
**Formula adjusted, standardized to suit performance parameters
*Chemically defined peptones
Directions
Suspend 29.0 gram in 1000 ml purified / distilled water. Heat if necessary to dissolve the medium completely. Mix well and dispense into sterile tubes or flasks as desired. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
Note: It is preferable to use freshly prepared medium, alternatively it should be boiled and cooled just once prior to use as on reheating, toxic oxygen radicles are formed.
Principle And Interpretation
Alternative Thioglycollate Medium is formulated as described in the N.I.H. memorandum (1). Alternative Thioglycollate HiCynth™ Medium is prepared by completely replacing animal based peptones with chemically defined peptones to avoid BSE/TSE/GMO risks associated with animal peptones. It is used for the sterility testing of certain biological products which are turbid or viscous and can be tested using Fluid Thioglycollate HiCynth™ Medium (MCD009)(2). Both the media have similar composition, except agar and resazurin that are not included in Alternative Thioglycollate HiCynth™ Medium. This deletion makes it suitable for sterility testing of viscous products. HiCynth™ Peptone No.3 serves as a source of nitrogen and carbon compounds, long chain amino acids and other essential nutrients. HiCynth™ Peptone No.5 serve as source of essential nutrients to the contaminants, if present. Dextrose serves as the energy source. Sodium chloride maintains the osmotic equilibrium of the medium whereas L-cystine, an amino acid, also serves as source of essential growth factors. Sodium thioglycollate and L-cystine lower the oxidation-reduction potential of the medium by removing oxygen to maintain a low Eh. Sodium thioglycollate also helps to neutralize the toxic effects of mercurial preservatives (3,4).
Type of specimen
Pharmaceutical: Sterility testing of viscous products.
Specimen Collection and Handling
For pharmaceutical products, follow appropriate techniques for sample processing in case of viscous materials as mentioned under sterility. (5)
Warning and Precautions
For professional use only. 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
- The tubes should not be reheated as frequent boiling leads to development of toxic products.
- Prior to use the medium should be boiled once to remove the absorbed oxygen.
- Before inoculation, the tubes should be brought to room temperature
- Further recovery on agar media and biochemical testing of pure colony is necessary 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 yellow homogeneous free flowing powder
Colour and Clarity of prepared medium
yellow coloured clear solution without any precipitate.
Reaction
Reaction of 2.9% 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 30-35°C for not more than 3 days.(#Incubated anaerobically).
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| #Clostridium sporogenes ATCC 19404 (00008*) | 50-100 | luxuriant |
| #Clostridium sporogenes ATCC 11437 | 50-100 | luxuriant |
| #Clostridium sporogenes NBRC 14293 | 50-100 | luxuriant |
| #Clostridium perfringens ATCC 13124 (00007*) | 50-100 | luxuriant |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | luxuriant |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | 50-100 | luxuriant |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | luxuriant |
| ^ Pseudomonas paraeruginosa ATCC 9027 (00026*) | 50-100 | luxuriant |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | luxuriant |
| Salmonella Abony NCTC 6017 (00029*) | 50-100 | luxuriant |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | luxuriant |
| #Bacteroides fragilis ATCC 23745 | 50-100 | luxuriant |
| #*Phocaeicola vulgatus ATCC 8482 | 50-100 | luxuriant |
Key: (*) Corresponding WDCM numbers, ^ Formerly known as Pseudomonas aeruginosa, * Formerly known as Bacteroides vulgatus ATCC 8482
Storage and Shelf Life
Store between 10-30°C in tightly closed container and the prepared medium at 15-30°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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (6,7).
| Product Name | Alternative Thioglycollate HiCynth™ Medium (Thioglycollate HiCynth™ Broth Alternative) |
|---|---|
| SKU | MCD010 |
| Product Type | HiCynth™ |
| Physical Form | Powder |
| Origin | Chemically defined (HiCynth™) |
| Packaging type | HDPE |
| References | 1. N.I.H. Memorandum, 1955: Culture Media for Sterility Tests, 4th |
| Customized Product Available | No |
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