Quality Control of Culture Media by Ecometric Method
Routine quality control of culture media is an essential “Good Laboratory Practice” necessary to maintain
the standards and performance of any culture media. More recently it is a requirement of many laboratory
accreditation schemes such as UKAS (United Kingdom Accreditation Service) and CLAS (Calibration
Laboratory Assessment Service, Canada) etc. A full quality control is recommended when a new batch of
dehydrated medium is introduced in the laboratory. This full quality control should be repeated every 3-4
months on opened containers. Each medium used in the laboratory should have its own quality control
protocol and the necessary organisms should be maintained.
In practice absolute measurements of growth are time-consuming or require sophisticated equipment
whilst colonial appearance is subjective and difficult to record in a culture inoculum. Comparative
methods are the most suitable ones for routine quality control of culture media. They can be used for
comparisons of growth and inhibition. The ecometric technique of Mossel, et al, 1983 is simple and gives
numerical readings that can form the basic records suitable for trend analysis. Both absolute growth index
(AGI) and relative growth index (RGI) can be obtained by this method.
Method:
1. Inoculate 5ml of Brain Heart Infusion Broth (TM 362) with a loopful of the recommended test
organism and incubate for 4 hours at an optimum temperature.
2. Prepare the selective culture medium to be tested as per the instructions and pour into sterile petri
plates. Divide the petri plate to be tested into quarters designated A, B, C and D as shown below:
Figure: Schematic diagram of the Ecometric method
3. Take 1 µl. inoculum of the incubated 4-hour culture and streak the test media plate, going from Al - Bl
- Cl - Dl - A2 - B2…etc. without flaming or recharging the loop and finish at D5.
4. Repeat this process with a control media plate. For a batch of dehydrated media new to the laboratory,
use suitable non-inhibitory and inhibitory organisms. For a new batch of media from the laboratory stock
of dehydrated media, use the same medium from the last batch to check consistency. Incubate both plates
for 18 hours.
5. After incubation, the last point should be noted in both test and control media plate at which growth
occurs, and record the segment and line, e.g. C4 or D5 etc. This is the end point and can be used to
calculate the absolute growth index (AGI) and relative growth index (RGI) of a medium. The AGI is
obtained by noting the end point in the Table below and it gives the AGI. e.g. if the end point is C4 then
the AGI is 75.
A1 = 5 B1 = 10 C1 = 15 D1 = 20
A2 = 25 B2 = 30 C2 = 35 D2 = 40
A3 = 45 B3 = 50 C3 = 55 D3 = 60
A4 = 65 B4 = 70 C4 = 75 D4 = 80
A5 = 85 B5 = 90 C5 = 95 D5 = 100
Table: Absolute Growth Index (AGI)
RGI is a comparison of the AGI of the test media plate and that of the control media plate. For example:
Plate A: Selective test media - End point C4, AGI = 75
Plate B: Non-selective control media - End point A5, AGI = 85
Using the formula RGI = AGI Test x 100
AGI Control
RGI = 75 x 100
85
= 88.24 %
Thus, for this particular organism the test media plate was 88.24% as efficient as the non-selective control
media plate. The performance of a selective agar can be thoroughly tested using an organism which has
been selected to check isolation and using another one which is selected to check inhibition. In order to
check efficiency, the RGI should be close to 100%, whereas for checking inhibition it should be closer to
0%. For example, Bile Esculin Azide Agar (TM038) is a very good selective medium as it will give high
RGI’s for faecal streptococci and achieve suppression of bacteria without inhibiting the organism which
is to be isolated. Through the comparisons of growth and inhibition by RGI, the user is therefore can
decide which medium is best for their use. After finding the best media, use that one as the benchmark to
measure the performance of new batches and to check that the efficiency of all media-making processes
are being maintained. It is important to note that the ecometric method is a simple technique by which
laboratories can check the media they are producing. However, it has to be performed with care, as simple
errors such as holding the inoculation loop at a different angle may introduce errors. This can be
overcome by using a Spiral Plater to inoculate the plates to ensure the plating method is identical for test
and control agars.
