Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate, Thane(W)-400604, MH, India
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HiEncap™ Mueller Hinton Agar
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SKU:
EC173CCL
For determination of susceptibility of microorganisms to antimicrobial agents.
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Intended Use:
Recommended for determination of susceptibility of microorganisms to antimicrobial agents.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| HM infusion B from $ | 300.000 |
| Acicaserm # | 17.500 |
| Starch | 1.500 |
| Agar | 17.000 |
| Final pH (at 25°C) | 7.3±0.1 |
**Formula adjusted, standardized to suit performance parameters
$ Equivalent to Beef, infusion from
# Equivalent to Casein acid hydrolysate
Directions
Each capsule contains 9.5 gms of medium. Suspend 1 capsule in 250 ml (4 capsules in 1000 ml) distilled or purified water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Mix well before pouring.
Note: The performance of this batch has been tested and standardized as per the current CLSI (formerly,NCCLS) document M6-Protocols for Evaluating Dehydrated Mueller Hinton Agar.
Principle And Interpretation
The Mueller Hinton formulation was originally developed as a simple, transparent agar medium for the cultivation of pathogenic Neisseria species (5). Other media were subsequently developed that replaced the use of Mueller Hinton Agar for the cultivation of pathogenic Neisseria species, but it became widely used in the determination of sulfonamide resistance of gonococci and other organisms. Mueller Hinton Agar is now used as a test medium for antimicrobial susceptibility testing (9). Mueller Hinton Agar is recommended for the diffusion of antimicrobial agents impregnated on paper disc through an agar gel as described in CLSI Approved Standard (10). Mueller Hinton Agar has been selected by the CLSI for several reasons:
- It demonstrates good batch-to-batch reproducibility for susceptible testing.
- It is low in sulfonamide, trimethoprim and tetracycline inhibitors.
- It supports the growth of most non-fastidious bacterial pathogens and
- Many data and much experience regarding its performance have been recorded (6).
Kirby-Bauer et al recommended this medium for performing antibiotic susceptibility tests using a single disc of high concentration (1). WHO Committee on Standardization of Susceptibility Testing has accepted Mueller Hinton Agar for determining the susceptibility of microorganisms because of its reproducibility (11). Mueller Hinton Agar with 5% sheep blood and Mueller Hinton Agar with Hemoglobin have been recommended for antimicrobial susceptibility testing of Streptococcus pneumoniae and Haemophilus influenzae.
HM infusion B and Acicase™ provide nitrogenous compounds, carbon, sulphur and other essential nutrients. Starch acts as a protective colloid against toxic substances present in the medium. Starch hydrolysis yields dextrose, which serves as a source of energy. These ingredients are selected for low thymine and thymidine content as determined by MIC values for Enterococcus faecalis with sulfamethoxazoletrimethoprim (SXT). Calcium and magnesium ion concentrations are adjusted to provide the amounts recommended by CLSI to give the correct MIC values with aminoglycosides and Pseudomonas aeruginosa (9).
The Kirby-Bauer procedure is based on agar diffusion of antimicrobial substances impregnated on paper discs. This method employs disc with a single concentration of antimicrobial agent and the zone diameters observed are correlated with minimum inhibitory concentration (MIC) values (2,5,9).
Mueller Hinton Agar is not appropriate for assay by disc diffusion method with slow growing organisms, anaerobes and capnophiles. With slow growing organisms, increased incubation may cause deterioration of diffusing antibiotic and produce unprecise readings (7).
Test Method Description
A standardized suspension of the organism is swabbed over the entire surface of the medium. Paper discs impregnated with specific amounts of antimicrobial agents are then placed on the surface of the medium, incubated and zones of inhibition around each disc are measured. The susceptibility is determined by comparing with CLSI standards (8). The various factors, which influence disc diffusion susceptibility tests, are agar depth, disc potency, inoculum concentration, pH of the medium and beta-lactamase production by test organisms (6,8).
Type of specimen
Clinical isolates
Warning and Precautions
In Vitro diagnostic 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
- This medium is recommended for susceptibility testing of pure cultures only.
- Inoculum density may affect the zone size. Heavy inoculum may result in smaller zones or too less inoculum may result in bigger zones.
- Fastidious organisms may not grow on this medium and may require supplementation of blood.
- Fastidious anaerobes may not grow on this medium.
- As antimicrobial susceptibility is carried with antibiotic disc, proper storage of the disc is desired which may affect the potency of the disc.
- Under certain circumstances, the in vitro results of antibiotic susceptibility may not show the same in vivo.
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
Gelatin capsule containing cream to yellow coloured granular media
Quantity
Each capsule contains 9.5 grams of medium sufficient for 250 ml medium
Gelling
Firm, comparable with 1.7% agar gel.
Colour and Clarity of prepared medium
Light amber coloured clear to slight opalscent gel froms in Petri plates.
Reaction
Reaction of 3.8% w/v aqueous solution at 25°C. pH: 7.3±0.1
pH
7.20-7.40
Cultural Response
Cultural characteristics observed after incubation at 35-37°C for 18-24 hours
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | >=70% |
| Haemophilus influenzae ATCC 49247 | 50-100 | good-luxuriant (on Mueller Hinton Chocolate Agar) | >=70% |
| Neisseria gonorrhoeae ATCC 49226 | 50-100 | luxuriant | >=70% |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | luxuriant | >=70% |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | luxuriant | >=70% |
| Enterococcus faecalis ATCC 29212 (00087*) | 50-100 | luxuriant | >=70% |
| Streptococcus pneumoniae ATCC 6305 | 50-100 | luxuriant (on Mueller Hinton Blood Agar) | >=70% |
| Escherichia coli ATCC 35218 | 50-100 | luxuriant | >=70% |
| Staphylococcus aureus ATCC 43300 (00211*) | 50-100 | luxuriant | >=70% |
Key: *Corresponding WDCM numbers.
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 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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (3,4).
| Product Name | HiEncap™ Mueller Hinton Agar |
|---|---|
| SKU | EC173CCL |
| Application/ Industry | Clinical |
| Product Type | HiEncap™ |
| Physical Form | Granulated Media in Gelatin Capsule |
| Origin | Animal |
| Packaging type | HDPE Jar |
| References | 1. Mueller J. H. and Hinton J., 1941, Proc. Soc. Exp. Biol. Med., 48:330. 2.National Committee for Clinical Laboratory Standards, 2000, Approved Standard: M7-A 5.Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that grow aerobically, 5th Ed., NCCLS, Wayne, Pa. 3.NCCLS Approved Standard: ASM-2, 1979, Performance Standards for Antimicrobic disc Susceptibility Tests, 2nd Ed., National Committee for Clin. Lab. Standards. 4.Bauer A. W., Kirby W. M., Sherris J. L. and Turck M., 1966, Am. J. Clin. Pathol., 45:493. 5.Present Status and Future Work, WHO Sponsored collaborative study, Chicago, Oct. 1967. 6.Ericsson H. M. and Sherris J. L., 1971, Acta Pathol. Microbiol., Scand. Sect B Suppl., 217:1. 7.National Committee for Clinical Laboratory Standards, 1986, Proposed Standards, M6-P, NCCLS, Villanova, Pa. 8.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williams and Wilkins, Baltimore |
| Customized Product Available | No |
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