Section: Microbiology Methods » Diagnostic Microbiology In Action

Simple Carbohydrate Fermentations

Content

Diagnostic Microbiology In Action
  Principles of Diagnostic Microbiology
    Primary Media for Isolation of Microorganisms
    Some Metabolic Activities of Bacteria
      Simple Carbohydrate Fermentations
      Starch Hydrolysis
      Production of Indole and Hydrogen Sulfide, and Motility
    Activities of Bacterial Enzymes
      The Activity of Urease
      The Activity of Catalase
      The Activity of Gelatinase
      The Activity of Deoxyribonuclease (Dnase)
      The Activity of a Deaminase
    Principles of Antigen Detection and Nucleic Acid Assays for Detection Identification of Microorganisms
      Antigen Detection Assays
      Enzyme Immunoassay (Eia)
      Nucleic Acid Detection Assays

Media for testing carbohydrate fermentation are often prepared as tubed broths, each tube containing a small inverted “fermentation” (or Durham) tube for trapping any gas formed when the broth is inoculated and incubated (see colorplate 18). Each broth contains essential nutrients, a specific carbohydrate, and a color reagent to indicate a change in pH if acid is produced in the culture (the broth is adjusted to a neutral pH when prepared). Organisms that grow in the broth but do not ferment the carbohydrate produce no change in the color of the medium, and no gas is formed. Some organisms may produce acid products in fermenting the sugar, but no gas, whereas others may form both acid and gas. In some cases, organisms that do not ferment the carbohydrate use the protein nutrients in the broth, thereby producing alkaline end products, a result that is also evidenced by a change in indicator color (see colorplate 19).


Purpose To distinguish bacterial species on the basis of simple carbohydrate fermentation
Materials Tubed phenol red glucose broth
Tubed phenol red lactose broth
Tubed phenol red sucrose broth
Slant cultures of Escherichia coli, Serratia marcescens, Pseudomonas aeruginosa, and Proteus vulgaris

Procedures
  1. Inoculate growth from each of the four cultures into separate tubes of each of the three carbohydrate broths. Be certain no bubbles are inside the Durham tubes before inoculation.
  2. Label each of the 12 inoculated tubes with the name of the carbohydrate it contains and the name of the bacterial culture.
  3. Incubate at 35°C for 24 hours.


Results
Record your results in the following table. Use these symbols to indicate specific changes observed in the broths.
  • A = acid production
  • K = alkaline color change
  • N = neutral (no change in color)
  • G = gas formation