Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use:
Recommended for the confirmation of the presumptive tests for coliforms from various samples.
Composition**
| Ingredients | g / L |
|---|---|
| HiVeg™ peptone | 10.000 |
| Lactose | 10.000 |
| Dipotassium hydrogen phosphate | 3.500 |
| Sodium sulphite | 2.500 |
| Agar | 12.000 |
| Final pH (at 25°C) | 7.5±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 38.0 grams in 1000 ml purified / distilled water. Add 4 ml of 10% Basic Fuchsin (FD059). Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. 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. Cool to 45-50°C. Mix well before pouring into sterile Petri plates.
Principle And Interpretation
Endo had first developed a culture medium for differentiation of lactose fermenters and lactose non-fermenters while inhibiting gram-positive bacteria (1). Inhibition of the later was achieved without the use of bile salts as was traditionally used. Endo was successful in inhibiting gram-positive bacteria on his medium by the incorporation of sodium sulphite and basic fuchsin. The resulting Endo Agar, also known as Fuchsin Sulphite and Infusion Agar, was used to isolate the typhoid bacilli. Many modifications of this media have been done over the years. Endo Agar is recommended by APHA as an important medium in the microbiological examination of water and wastewater, dairy products and foods (2,3,4). Endo Agar is used to confirm the detection and enumeration of coliform bacteria following presumptive test of drinking water. It is also used for the detection and isolation of coliforms and fecal coliforms from milk, dairy products and food. Endo HiVeg™ Agar Base, Modified 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™ peptone which provide nitrogen, carbon, vitamins and minerals required for bacterial growth. Sodium sulphite and basic fuchsin (FD) has inhibitory effect on gram-positive microorganisms. Lactose fermenting coliforms produce aldehyde and acid. The aldehyde in turn liberates fuchsin from the fuchsin-sulphite complex, giving rise to a red colouration of colonies. With Escherichia coli, this reaction is very pronounced as the fuchsin crystallizes, exhibiting a permanent greenish metallic lustre (fuchsin lustre) to the colonies.
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 (2). 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
Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.2% Agar gel
Colour and Clarity of prepared medium
After addition of FD059: Orangish pink coloured, After addition of FD059: clear to slightly opalescent gel with fine precipitate forms in Petri plates.
Reaction
Reaction of 3.8% w/v aqueous solution at 25°C. pH: 7.5±0.2
pH
7.30-7.70
Cultural Response
Cultural characteristics observed with added Basic fuchsin (FD059) 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 (5,6).
| Product Name | Endo HiVeg™ Agar Base |
|---|---|
| SKU | MV1077 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Endo, 1904, Zentralbl. Bakteriol., Abt. I. Orig., 35:109. |
| Customized Product Available | No |
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