Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Dey-Engley Neutralizing Agar Plate recommended in disinfectant testing where neutralization of the chemical is important for determining its bactericidal activity.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Tryptone | 5.000 |
| Yeast extract | 2.500 |
| Dextrose (Glucose) | 10.000 |
| Sodium thiosulphate | 6.000 |
| Sodium thioglycollate | 1.000 |
| Sodium bisulphite | 2.500 |
| Lecithin | 7.000 |
| Polysorbate 80 | 5.000 |
| Bromocresol purple | 0.020 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.6±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.
Principle And Interpretation
Dey-Engley Neutralizing Agar Plate is formulated as per the procedure described by Engley and Dey (1). A strongly bacteriostatic substance inhibits the growth and reproduction of bacteria without killing them. These bacteria hold the ability to cause infection under favorable conditions. Dey-Engley Neutralizing Agar neutralizes a broad spectrum of antiseptics and disinfectants including quaternary ammonium compounds, phenolic, iodine and chlorine preparations, mercurial, formaldehyde and glutaraldehyde (2).
Tryptone provide nitrogen and carbon source, long chain amino acids, vitamins and other essential nutrients. Dextrose is an energy source. Yeast extract is also a rich source of vitamin B-complex. The present formulation incorporate neutralizing substances for almost all the active products used as antiseptics and disinfectants. Sodium bisulfite neutralizes aldehydes; sodium thioglycollate neutralizes mercurial; sodium thiosulfate neutralizes iodine and chlorine (1); lecithin neutralizes quaternary ammonium compounds; and polysorbate 80, a non-ionic surface-active agent, neutralizes substituted phenolic (1,2,3,4). Bromocresol purple is an indicator for dextrose utilization. Due to the high concentration of lecithin in the broth medium, turbidity cannot be used to detect growth. Therefore, bromocresol purple and dextrose are added to the medium. Those organisms that ferment dextrose will turn the medium from purple to yellow (1).
Neutralization Test: Growth in Neutralizing Broth and no growth in Neutralizing Broth Base indicate neutralization of disinfectant. To check bactericidal activity, both broth tubes are inoculated on D/E Neutralizing Agar. Positive growth from negative tubes of Neutralizing Broth Base indicates bacteriostatic substance while negative growth indicates a bactericidal disinfectant. All positive tubes should show growth on Dey-Engley Neutralizing Agar. The control disinfectants used in test procedure are 2% chlorine, 2% formaldehyde, 1% glutaraldehyde, 2% iodine, 2% phenol, 1/750 quaternary ammonium compounds, 1/1000 mercurial etc.
Type of specimen
Food and dairy samples; Environmental samples.
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (4,5).
For environmental samples follow appropriate techniques for handling specimens as per established guidelines (6,7).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
Read the label before opening the pack. 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
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
- It is recommended to store the plates at 24-30°C to avoid minimum condensation.
- Further biochemical identification is necessary 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
Sterile Dey/Engley Neutralizing Agar in 90 mm Sterile disposable plate with smooth surface and absence of black
particles/ cracks/bubbles (Gamma Irradiated, Triple Pack)
Colour of medium
Purple coloured medium
Quantity of medium
30 ml of medium in 90mm disposable plate
pH
7.40-7.80
Dose of irradiation (Kgy)
13.00-20.00
Sterility Check
Passes release criteria
Cultural Response
Cultural characteristics observed after an incubation at 30 - 35°C for <=3 days in case of bacteria and <=5 days in case of
fungi
| Organism (ATCC) | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Grovth at 30-35°C for < = 3 days | |||
| **Bacillus spizizenii ATCC 6633 (00003*) |
50-100 | luxuriant | >=70% |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) |
50-100 | luxuriant | >=70% |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) |
50-100 | luxuriant | >=70% |
| Escherichia coli ATCC 25922 (00013*) |
50-100 | luxuriant | >=70% |
| Escherichia coli ATCC 8739 (00012*) |
50-100 | luxuriant | >=70% |
| Pseudomonas aeruginosa ATCC 27853 (00025*) |
50-100 | luxuriant | >=70% |
| ^Pseudomonas paraaeruginosa ATCC 9027 (00026*) |
50-100 | luxuriant | >=70% |
| Salmonella Abony NCTC 6017 (00029*) |
50-100 | luxuriant | >=70% |
| $Kocuria rhizophila ATCC 9341 |
50-100 | luxuriant | >=70% |
| ^^Bacillus spizizenii ATCC 19659 |
50-100 | luxuriant | >=70% |
| Growth at 20-25°C for < = 5days | |||
| Candida albicans ATCC 10231 (00054*) |
50-100 | luxuriant | >=70% |
| Candida albicans ATCC 2091 (00055*) |
50-100 | luxuriant | >=70% |
| # Aspergillus brasiliensis ATCC 16404 (00053*) |
50-100 | luxuriant | >=70% |
| ##Penicillium rubens ATCC11709 |
50-100 | luxuriant | >=70% |
Key-*Corresponding WDCM numbers
^ Formerly known as Pseudomonas aeruginosa
# Formerly known as Aspergillus niger
##Formerly known as Penicillium chrysogenum
**Formerly known as Bacillus subtilis subsp. spizizenii
$ Formerly known as Micrococcus luteus
^^Formerly known as Bacillus subtilis
Storage and Shelf Life
On receipt store between 20-30°C. Use before expiry date on the label. 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 | Dey Engley Neutralizing Agar Plate (γ-irradiated) (Triple Pack) |
|---|---|
| SKU | MP186GT |
| Customized Product Available | No |
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