HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended Use
Recommended for determination of susceptibility of Streptococcus species to antimicrobial agents.
Composition**
| Ingredients | g/L |
|---|---|
| HM infusion from 300 g # | 2.000 |
| Acicase ## | 17.500 |
| Starch | 1.500 |
| Agar | 17.000 |
| Sheep Blood | 50.000 ml |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef, infusion from
## Equivalent to Casein acid hydrolysate
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.
Principle And Interpretation
The Mueller Hinton formulation was originally developed as a simple, transparent agar medium for the cultivation of pathogenic Neisseria species (1). 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 (2). Mueller Hinton Agar is recommended for the diffusion of antimicrobial agents impregnated on paper disc through an agar gel as described in CLSI and EUCAST approved Standard (3,4).
Kirby-Bauer et al recommended this medium for performing antibiotic susceptibility tests using a single disc of high concentration (5). WHO Committee on Standardization of Susceptibility Testing has accepted Mueller Hinton Agar for determining the susceptibility of microorganisms because of its reproducibility (6).
HM infusion B from 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 sulfamethoxazole trimethoprim (SXT).
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 (7,8,9). 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 & EUCAST standards (2,4).
Type of specimen
Clinical samples Isolated microorganisms from urine, stool, pus etc. According to CLSI & EUCAST, use only pure cultures. Clinical swab samples may be used directly, but should be confirmed with repeated test with pure cultures.
Specimen Collection and Handling
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 (10).
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 (3,10). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precaution
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
- 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.
- 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.
- Each lot of the medium has been tested for the organisms specified on the COA.
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 Mueller Hinton Agar Plate w/ 5% Sheep Blood in 90 mm disposable plates with convex surface and absence of black particles/cracks/bubbles.
Colour
Red coloured medium
Quantity of medium
25 ml of medium in 90 mm disposable plate
pH
7.10-7.50
Sterility Check
Passes release criteria
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours with added 5% w/v sterile sheep blood.
| Organism | Growth | Recovery | Haemolysis | Clindamycin CD 2 mcg | Erythromycin E 15mcg | Vancomycin VA 30 mcg |
|---|---|---|---|---|---|---|
| Streptococcus pyogenes ATCC 19615 | luxuriant | >=70% | beta | |||
| Streptococcus pneumoniae ATCC 49619 | luxuriant | >=70% | alpha | 19-25 mm | 25-30 mm | 20-28 mm |
Storage and Shelf Life
On receipt store between 2-8°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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (12,13).
| Product Name | Mueller Hinton Agar Plate w/ 5% Sheep Blood |
|---|---|
| SKU | MP1806 |
| Customized Product Available | No |
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