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Endo HiVeg™ Agar w/ NaCl
Intended Use
Recommended for detection and isolation of pathogenic enteric bacilli.
Composition
| Ingredients | g/L |
|---|---|
| HiVeg™ special peptone | 8.000 |
| Lactose | 10.000 |
| Sodium chloride | 3.000 |
| Dipotassium hydrogen phosphate | 2.000 |
| Sodium sulphite | 2.500 |
| Basic Fuchsin | 0.200 |
| Agar | 12.000 |
| Final pH (at 25°C) | 7.5±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Suspend 37.7 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. Mix well and pour into sterile Petri Plates. If the solidified culture medium is somewhat too red, then to remove the colour, add a few drops (Max. 1 ml/litre) of a freshly prepared 10% Sodium sulphite solution and boil.
Principle And Interpretation
Endo Agar was developed by Endo to differentiate gram-negative bacteria on the basis of lactose fermentation, while inhibiting gram-positive bacteria (1). Endo was successful in inhibiting gram-positive bacteria on this medium by the incorporation of sodium sulphite and basic fuchsin Endo Agar w/ NaCl is prescribed in the regulations for the execution of the German Meat Inspection Law (2). Endo HiVeg™ Agar w/ NaCl is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks. The medium contains HiVeg™ special peptone special which provide nitrogen, carbon, vitamins and minerals required for bacterial growth. Sodium sulphite and basic fuchsin inhibits most of the gram-positive bacteria. Lactose fermenting Escherichia coli and coliforms produce aldehyde and acid. The aldehyde liberates fuchsin from the fuchsin-sulphite complex and colonies of lactose fermenters appear dark red. Non-lactose fermenters show colourless colonies. With Escherichia coli, this reaction is very pronounced as the fuchsin crystallizes, exhibiting a permanent greenish metallic luster (fuchsin luster) to the colonies. Medium should be stored away from light to avoid photo-oxidation.
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 (3,4). For water samples follow appropriate techniques for sample collection, processing as per guidelines and local standards (5). 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
- Besides Enterobacteriaceae, other gram negative bacteria and yeasts may also grow.
- Avoid exposure of the medium to light, as it may lead to photo oxidation and decrease productivity of the medium.
- Overheating of the medium must be avoided, as it may destroy the productivity of the medium.
- Further biochemical tests must 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: Light pink to purple homogeneous free flowing powder
Gelling: Firm, comparable with 1.2% Agar gel
Colour and Clarity of prepared medium: Orangish pink coloured, clear to slightly opalescent gel with fine precipitate forms in Petri plates.
Reaction: Reaction of 3.77% w/v aqueous solution at 25°C. pH: 7.5±0.2
pH: 7.30-7.70
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony |
|---|---|---|---|---|
| **Bacillus spizizenii ATCC 6633 (00003*) | >=104 | inhibited | 0% | |
| Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | good-luxuriant | >=50% | pink |
| Enterococcus faecalis ATCC 29212 (00087*) | 50-100 | none-poor | <=10% | pink, small |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | good-luxuriant | >=50% | pink to rose red with metallic sheen |
| Klebsiella pneumoniae ATCC 13883 (00097*) | 50-100 | good-luxuriant | >=50% | pink, mucoid |
| ##Proteus hauseri ATCC 13315 | 50-100 | good-luxuriant | >=50% | colourless to pale pink |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | good-luxuriant | >=50% | colourless, irregular |
| Salmonella Typhi ATCC 6539 | 50-100 | good-luxuriant | >=50% | colourless to pale pink |
| Shigella sonnei ATCC 25931 | 50-100 | good-luxuriant | >=50% | colourless to pale pink |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=104 | inhibited | 0% | |
| Enterobacter cloacae ATCC 13047 (00083*) | 50-100 | good | 40-50% | pink |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | good-luxuriant | >=50% | colourless |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | good-luxuriant | >=50% | colourless |
| Shigella flexneri ATCC 12022 (00126*) | 50-100 | good-luxuriant | >=50% | colourless |
Key: (*) Corresponding WDCM numbers **- Formerly known as Bacillus subtilis subsp. spizizenii ## - Formerly known as Proteus vulgaris.
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 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 | Endo HiVeg™ Agar w/ NaCl |
|---|---|
| SKU | MV1258 |
| Customized Product Available | No |

