Mueller Hinton Agar

Mueller Hinton Agar: A Comprehensive Guide for Microbiological Applications

Published: 30th Jan 2024, 11:28

Mueller Hinton Agar (MHA) is a non-selective solid media widely used in microbiology for various applications, with its primary focus on antimicrobial susceptibility testing. In this blog, we will explore the composition, principle, uses, and quality control of MHA, shedding light on its significance in the field of microbiology.

Composition of Mueller Hinton Agar

Mueller Hinton Agar is composed of carefully selected ingredients to provide a suitable environment for the growth of a variety of bacteria. The key components and their concentrations are outlined in the table below:

Beef infusion300
Casein acid hydrolysate17.500


  • Beef infusion and Casein acid hydrolysate supply nitrogenous compounds, carbon, sulphur, and essential nutrients.
  • Starch acts as a protective colloid against toxic substances, ensuring a conducive environment for bacterial growth.
  • Starch hydrolysis yields dextrose, serving as a source of energy.

Uses of Mueller Hinton Agar

  1. Antimicrobial Susceptibility Testing

The primary and most crucial application of MHA is in antimicrobial susceptibility testing. It has become the standard medium for the Kirby Bauer method, with its performance specified by the Clinical and Laboratory Standards Institute (CLSI).

  1. Cultivation of Neisseria

Initially developed for the cultivation of pathogenic Neisseria species, MHA is still being used for the cultivation of Neisseria.

  1. Food Testing and Other Bacteriological Procedures

MHA is specified in the FDA Bacteriological Analytical Manual for food testing and commonly used procedures on aerobic and facultative anaerobic bacteria.

Why MHA for Antibiotic Susceptibility Testing?

Non-selective and non-differential: 

  • MHA supports the growth of almost all organisms plated on it, making it a versatile medium for susceptibility testing.

Starch Content: 

  • Starch in MHA absorbs toxins released from bacteria, preventing interference with antibiotics.
  • Mediates the rate of antibiotic diffusion through the agar.

Loose Agar:

  • The loose agar structure allows for better diffusion of antibiotics, resulting in more accurate zone of inhibition measurements.

Batch-to-Batch Reproducibility:

  • MHA demonstrates good reproducibility for susceptibility testing, ensuring reliable results across different batches.

Low in Inhibitors:

  • MHA is low in sulfonamide, trimethoprim, and tetracycline inhibitors, minimizing interference in susceptibility testing.

Quality Control of MHA

To ensure the reliability of MHA in microbiological applications, positive controls can be employed:

  • Positive Controls:
    • Escherichia coli: Good growth with pale straw-colored colonies.
    • Pseudomonas aeruginosa: Good growth with straw-colored colonies.
    • Staphylococcus aureus: Good growth with cream-colored colonies.

By regularly performing quality control measures, laboratories can maintain the effectiveness and consistency of Mueller Hinton Agar for antimicrobial susceptibility testing.

Mueller Hinton Agar remains a cornerstone in microbiology laboratories, especially in antimicrobial susceptibility testing. Its well-balanced composition, coupled with ease of use and reliable performance, makes it an indispensable tool for researchers and healthcare professionals alike. As technology advances, MHA continues to play a crucial role in the accurate determination of bacterial susceptibility to antimicrobial agents, contributing to the ongoing battle against infectious diseases.

At TM Media , we take pride in offering high-quality Mueller Hinton Agar (MHA) and its variants, meticulously designed for precise antimicrobial susceptibility testing and microbial cultivation. Our MHA formulations, including those with 5% sheep blood and hemoglobin, adhere to CLSI standards, ensuring reliability and reproducibility. Trust TM Media for all your microbiological needs, where excellence meets your laboratory requirements. Visit our website for more information.

More Blogs