A Simple Guide to Microbiological Culture Media

Hand Holds Petri Dish with Bacteria Culture stock

Different microorganisms have different nutritional needs, but all of them require carbon, nitrogen, and trace elements to grow. In laboratories, we use culture media to provide these nutrients.

There are two main types of culture media:

  1.     1. Chemically defined or synthetic media: These have known exact ingredients.
  2.     2. Non-synthetic media: These have some unknown components.

Most microbiological tests use non-synthetic media. Synthetic media are usually only used for special experiments or research.

Some microbes can grow in simple media containing just nitrogen or nitrates. Others need more complex ingredients like peptone (a protein digest) as a nitrogen source. Most microbes found in clinical settings need even more complex compounds to grow well.

Culture media can be categorized into several types based on their purpose or function:


I. Enriched Media

These media are supplemented with special nutrients such as blood, serum, or animal/plant extracts to support the growth of fastidious bacteria. Examples include:

  •     ✅Blood Agar Plate (BAP): Supports the growth of Streptococcus pneumoniae.
  •     ✅Brucella Agar (or AnBAP): Supports anaerobic growth.
  •     ✅Chocolate Agar: Supports the growth of Haemophilus influenzae. Other enriched media include Tryptic Soy Agar (TSA), Tryptic Soy Broth (TSB), TGEA, Plate Count Agar (PCA), Nutrient Agar (NA), LB Agar, and Mueller-Hinton (M-H) Agar.


II. Selective Media

These media contain specific chemical substances that inhibit the growth of one group of bacteria while allowing another group to multiply. Examples include:

  •     ✅Eosin Methylene Blue (EMB) Medium and MacConkey Medium: Allow growth of Gram-negative rods (e.g., Escherichia coli) but inhibit Gram-positive cocci (e.g., staphylococci).
  •     ✅Antibiotic-containing isolation media (e.g., CAN): Selectively inhibit certain microorganisms (enterobacteria) while allowing specific microorganisms (staphylococci) to grow well.


III. Differential Media

These media contain special chemical substances that allow different bacteria to display distinct characteristics. For example:

  •     ✅MacConkey Medium: Lactose-fermenting Gram-negative rods form pink colonies, while non-lactose fermenters form colorless colonies.
  •     ✅EMB Agar: Lactose fermenters form deep purple or blue-black colonies, while non-fermenters form colorless colonies. Both MacConkey and EMB agar serve as differential and selective media. Blood Agar Plate (BAP) can differentiate hemolytic and non-hemolytic bacteria, making it both an enriched and differential medium.


IV. Enrichment Media

These media contain special components that favor the growth of desired bacterial species while inhibiting others. They are typically used to isolate slow-growing or less abundant species from samples. Examples include:

  •     ✅GN Broth and Selenite Brilliant Green Broth: Used for fecal sample microbial examination, encouraging the growth of Salmonella while suppressing common bacteria like E. coli.
  •     ✅Alkaline Peptone Water: Promotes the growth of Vibrio cholerae while inhibiting other bacteria due to its high pH.


V. Media for Identifying Bacterial Characteristics

a) Carbohydrate Fermentation Test Media:

  •     - Examples: Bromocresol Purple Broth w/1% Lactose, CTA w/1% Carbohydrate, Phenol Red Broth w/1% Carbohydrate, Andrade's Broth w/1% Carbohydrate, O/F Broth w/1% Carbohydrate.

b) Substrate Utilization Test Media:

  •     - Examples: Simmons Citrate Agar, Acetate Agar.

c) Enzyme-Producing Ability Test Media:

  •     - Examples: DNase Test Medium, Christensen Urea Agar, Ornithine or Lysine Decarboxylase Medium.

d) Special Metabolite Detection Media:

  •     - Examples: Methyl Red, VP Medium.

e) Motility Test Media:

  •     - Example: Motility Test Medium.

f) Chromogenic Agar:

  •     - Example: CHROMagar series for various pathogens.


VI. Preservation and Transport Media

These media are used to maintain the viability and physiological characteristics of pure cultures. Examples include:

  •     ✅Stock Medium: For preserving common enterobacteria.
  •     ✅GermBank Preservation Device: For preserving various pathogens.

Transport media are crucial for clinical specimens to maintain microbial populations without growth or reduction before culturing. Examples include:

  •     ✅Aerobic CultureSwab (containing Stuart, Amies, or Cary-Blair medium)
  •     ✅Anaerobic CultureSwab or Anaerobic Transtube
  •     ✅Buffered Glycerol-Saline Base (for Shigella spp. isolation from feces)
  •     ✅Campy-transtube (for Campylobacter spp. isolation from feces)

Advantages of transport media (using Amies and Stuart as examples):

  1.     1. No nutrients - microorganisms don't multiply.
  2.     2. Buffered semi-solid agar - maintains swab moisture and appropriate pH.
  3.     3. Contains sodium thioglycollate as a reducing agent.
  4.     4. Activated charcoal (if needed) - removes toxic metabolites but may interfere with smear observation.