Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for urine bacteria and Enterococci screening.
Composition
| Ingredients | g / L |
|---|---|
| MacConkey Agar w/ 0.15% Bile Salts, CV and NaCl | |
| Gelatin peptone | 17.000 |
| Tryptone | 1.500 |
| Peptone | 1.500 |
| Lactose | 10.000 |
| Bile salts | 1.500 |
| Sodium chloride | 5.000 |
| Neutral red | 0.030 |
| Crystal violet | 0.001 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.1±0.2 |
| C.L.E.D. Agar w/Bromo Thymol Blue | |
| Peptone | 4.000 |
| Tryptone | 4.000 |
| HM Peptone B# | 3.000 |
| Lactose | 10.000 |
| L-Cystine | 0.128 |
| Bromothymol blue | 0.020 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.3±0.2 |
| Bile Esculin Agar | |
| Peptone | 5.000 |
| HM peptone B # | 3.000 |
| Bile. | 40.000 |
| Esculin | 1.000 |
| Ferric citrate | 0.500 |
| Agar | 15.000 |
| Final pH (at 25°C) | 6.7±0.2 |
**Formula adjusted, standardized to suit performance parameters
#- Equivalnet to Beef extract
#- Equivalent to Oxgall
Directions
- Surfaces: Loosen cap and remove HiDip™ slide from container taking care not to touch agar surfaces. Check for dehydration or contamination. Gently lower the slides and press agar to touch the test surface by bending the scull around the hinge line. Apply even and firm pressure for 15-20 seconds. Take care not to smudge agar over the test surface. Repeat procedure using the second agar surface on an area adjacent to the initial test side. Return the slide to the container and close tightly. Incubate in an up right position at indicated temprature.
- Liquids: Loosen cap and remove the HiDip™ slide from container. Check for dehydration or contamination.Dip slide into test fluid for upto 15-20 seconds so that agar surface becomes totally covered. (In case of inadequate liquid sample availability, pour sample over the surface of the slide). Allow to drain. Tab it gently to remove excess fluid from surface. Return the slide to the container and close tightly. Incubate in an upright position at indicated temprature. Label the container for sample number, source, date and time etc. for reference.
Principle And Interpretation
MacConkey Agar is a differential medium for the selection and recovery of the Enterobacteriaceae and related enteric gram-negative bacilli. This medium is prepared in accordance with USP and contains crystal violet, Nacl and bile salts. It is very selective and suppresses growth of a number of gram-positive bacteria including Staphylococci.
C.L.E.D.Agar is recommended for isolation, enumeration and identification of urinary pathogens on the basis of lactose fermentation.
Bile Esculin Medium is in accordance with APHA for the presumptive identification of Group D Streptococci. ISO Committee has also suggested it for the isolation and identification of Yersinia enterocolitica. Esculin hydrolyzers, hydrolyze the glycoside esculin to esculetin and dextrose. Esculetin reacts with ferric citrate to form a black-brown complex.
Type of specimen
Clinical sample : Urine; Food and dairy samples, Water samples.
Specimen Collection and Handling
Refer Directions.
Warning and Precautions
In Vitro diagnostic use only. 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 clinical specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
MacConkey Agar:
- Though the medium is recommended for selective isolation, further biochemical and serological testing must be carried out for further confirmation.
C.L.E.D.Agar
- This medium is recommended for urine infection. Low urine count may be a result of antibiotic therapy, low pH of urine.
- Recovery depends on the urine count.
- Inoculate the medium immediately after urine collection.
Bile Esculin Agar
- This medium is general purpose medium and may not support the growth of fastidious organisms.
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
The HiDip™ slide containing combination of sterile MacConkey Agar Medium, CLED Agar Medium and Bile Esculin Agar Medium on separate individual surfaces.
Colour
Colour of MacConkey Agar
Red coloured gel with puplish tinge
Colour of CLED Agar
Green coloure gel.
Colour of Bile Esculin Agar
Amber coloured gel with slightly bluish tinge
Quantity of medium
1.5ml of medium per surface
Sterility Check
Passes release criteria
pH of MacConkey Agar
pH Range: 6.90-7.30
pH of CLED Agar
pH Range: 7.10- 7.50
pH of Bile Esculin Agar
pH Range: 6.50-6.90
Cultural Response
Cultural characteristics observed after an incubat ion at 35-37° C for 24-72 hours.
| Organism | Growth | Colour of colony |
|---|---|---|
| MacConkey Agar | ||
| Escherichia coli ATCC 25922 (00013*) | luxuriant | pink to red with bile precipitate |
| # Klebsiella aerogenes ATCC 13048 (00175*) | luxuriant | pink to red |
| Enterococcus faecalis ATCC 29212 (00087*) | fair to good | Pale pink to red |
| Staphylococcus aureus subsp.aureus ATCC 25923 (00034*) | inhibited | - |
| Salmonella Enteritidis ATCC 13076 (00030*) | luxuriant | colourless |
| Salmonella Paratyphi B ATCC 8759 | luxuriant | colourless |
| Salmonella Typhi ATCC 6539 | luxuriant | colourless |
| ## Proteus hauseri ATCC 13315 | luxuriant | colourless |
| Shigella flexneri ATCC 12022 (00126*) | fair to good | colourless |
| CLED Agar | ||
| Escherichia coli ATCC 25922 (00013*) | good-luxuriant | yellow, opaque, centre slightly deeper yellow |
| Enterococcus faecalis ATCC 29212 (00087*) | good-luxuriant | slight yellowish or greenish |
| ## Proteus hauseri ATCC 13315 | good-luxuriant | blue |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | good-luxuriant | deep yellow |
| Salmonella Typhi ATCC 6539 | good-luxuriant | bluish |
| Klebsiella pneumoniae ATCC 13883 (00097*) | good-luxuriant | yellow to whitish blue |
| Bile Esculin Agar | ||
| Enterococcus faecalis ATCC 29212 (00087*) | luxuriant | positive reaction, blackening of medium around the colony |
| Proteus mirabilis ATCC 25933 | luxuriant | negative reaction |
| Streptococcus pyogenes ATCC 19615 | none-poor | negative reaction |
Key :- * Corresponding WDCM numbers
# Formerly known as Enterobacter aerogenes
## Formerly known as Proteus vulgaris
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
Used HiDip™ slides should be handled carefully, as it contains live microorganisms. These slides can be best disposed off either by or by immersing in a suitable disinfectant solution (i.e. dettol, phenyl etc.) over night or by autoclaving them after loosening the cap.
| Product Name | HiDip™ Mac-CLED-Bile Esculin Medium |
|---|---|
| SKU | HD003 |
| Customized Product Available | No |
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