Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Eijkman Lactose HiVeg™ Broth
For detection and differentiation of Escherichia coli from other coliforms on the basis of their ability to grow and liberate gas from lactose.
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Intended Use
Recommended for the detection and differentiation of Escherichia coli from other coliform organisms on the basis of their ability to grow and liberate gas from lactose.
Composition
| Ingredients | g/L |
|---|---|
| HiVeg® hydrolysate No. 1 | 15.000 |
| Lactose | 3.000 |
| Dipotassium hydrogen phosphate | 4.000 |
| Potassium dihydrogen phosphate | 1.500 |
| Sodium chloride | 5.000 |
| Final pH (at 25°C) | 6.8±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 28.5 grams in 1000 ml purified / distilled water. For examination of 10 ml portions of water samples, use 57 grams per 1000 ml distilled water. Heat if necessary to dissolve the medium completely. Dispense into tubes with inverted Durhams fermentation tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C.
Principle And Interpretation
Coliform organism is a term used to designate the lactose-fermenting Enterobacteria such as Escherichia coli and Enterobacter. Enterobacteriaceae forms a large group of gram-negative bacteria that inhabit intestinal tract of warm-blooded animals. Therefore they constitute the major microbial flora of human faeces. Since coliforms are readily isolated and identified, they are used as indicator organisms to check faecal contamination of food, water and other samples (1).
E.coli is one of the common organisms involved in gram-negative sepsis and endotoxin-induced shock (2). Eijkman (3) described a method for selective isolation of E. coli from faeces of warm-blooded and cold-blooded animals. This method had limitations due to the inability to obtain growth after subculturing from positive tubes incubated at 46°C, as acidity and high temperature resulted in death of the culture within 24-48 hours. Perry and Hajna (4) modified Eijkmans original method by decreasing carbohydrate content and adding a phosphate buffer enabling to subculture E. coli after incubation at 46°C for 96 hours or longer where pH was 5.6 unlike 4.5 of Eijkman Medium. Perry (5) modified Eijkman Medium using lactose for isolation of E. coli. This medium can also be used for bacteriological examination in water filtration control work (6). MacConkey HiVeg® Agar w/ CV,NaCl, 0.005% NR and 1.5% Agar is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks.
HiVeg® hydrolysate No. 1 and lactose in the medium are the energy and the carbon sources respectively. E. coli ferment lactose to form acid and gas. The gas produced gets trapped in the form of gas bubbles in the inverted Durhams tubes. Phosphates buffer the medium whereas sodium chloride helps to maintain the osmotic equilibrium of the medium.
Type of specimen
Water samples.
Specimen Collection and Handling
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards(7). 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
- This method had limitations due to the inability to obtain growth after subculturing from positive tubes incubated at 46°C, as acidity and high temperature resulted in death of the culture within 24-48 hours.
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
Light yellow coloured, clear solution without any precipitate
Reaction
Reaction of 2.85% w/v aqueous solution at 25°C. pH: 6.8±0.2
pH
6.60-7.00
Cultural Response
Cultural characteristics observed after an incubation at 45.5 to 46°C for 24 - 48 hours.
| Organism | Inoculum (CFU) | Growth | Gas |
|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | positive reaction |
| # Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | poor | negative reaction |
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 15-30°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order 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 (8,9).
| Product Name | Eijkman Lactose HiVeg™ Broth |
|---|---|
| SKU | MV086 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg), Lactose |
| Packaging type | HDPE |
| References | 1. Norton C. F., 1940, Microbiology, 2nd Ed., Addison Wesley Publishing Company. 2.Koneman E. W., Allen S. D., Janda W. M., Schreckenberger P. C., Winn W. C. Jr., 1992, Colour Atlas and Textbook ofDiagnostic Microbiology, 4th Ed, J. B. Lippincott Company. 3.Eijkman, 1904, Centr. Bakt., 11th Abst., 37:742. 4.Perry C. A., 1939, Food Research, 4:381. 5.Perry C. A. and Hajna A. A., 1933, J. Bacteriol., 26:419. 6.Standard Methods for the Examination of Water and Wastewater, 11th Ed., 1960, APHA, N.Y. |
| Customized Product Available | No |
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Eijkman Lactose Broth
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