Section II

Dehydrated Culture Media, cont.

XII. Limitations of the Procedure potential significance in a clinical specimen or industrial

Detergent residues can compromise media. After washing
glassware, check for alkali or acid residue with a few drops
of bromthymol blue pH indicator (yellow is acidic; blue is
alkaline).
Quantities of media in excess of one liter may require an
extended autoclave time to achieve sterilization. Longer
sterilization cycles can cause nutrient concentration changes
and generation of inhibitory substances.
sample. The agents in selective media may inhibit some strains
of the desired species or permit growth of a species they were
designed to inhibit, especially if the species are present in large
numbers in the specimen/sample. Cultures of specimens/samples
grown on selective media should, therefore, be compared with
specimens/samples cultured on nonselective media to obtain
additional information and help ensure recovery of potential
pathogens and other significant organisms.

Since the nutritional requirements of organisms vary, a single XIII. Reference

medium is rarely adequate for detecting all organisms of 1. U.S. Public Health Service, Centers for Disease Control and Prevention, and National Institutes of
Health. 2007. Biosafety in microbiological and biomedical laboratories, 5th ed. HHS Publication
No. (CDC) 93-8395. U.S. Government Printing Office, Washington, D.C.

Problem A B C D E F G H Other Causes

Abnormal color of medium • • •
Incorrect pH • • • • • • • Storage at high temperature
Hydrolysis of ingredients
pH determined at wrong temperature
Atypical precipitate • • • • • •
Incomplete solubility • Inadequate heating
Inadequate convection in a too small
flask
Darkening or carmelization • • • •
Toxicity • • Burning or scorching
Trace substances (Vitamins) • Airborne or environmental sources
of vitamins
Loss of gelation property • • • • Hydrolysis of agar due to pH shift
Not boiling medium
Loss of nutritive value or • • • • • • • Burning or scorching
selective or differential Presence of strong electrolytes, sugar
properties solutions, detergents, antiseptics,
metallic poisons, protein materials or
other substances that may inhibit the
inoculum
Contamination • • • Improper sterilization
Poor technique in adding enrichments
and pouring plates
Not boiling agar containing medium

Key
A Deteriorated Dehydrated Medium D Incorrect Weighing G Repeated Remelting
B Improperly Washed Glassware E Incomplete Mixing H Dilution by a Too Large Inoculum
C Impure Water F Overheating

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