ISO 11290-2 (2017) medium: ALOA

Media control Food chain

Microorganisms and performance criteria for testing selective media 'ALOA' for Enumeration of Listeria monocytogenes used in Food chain microbiology according to ISO 11290-2.

  • Quantitative control of Productivity

    • Best Practice ALOA

    Quantitative control of Productivity solid medium

    1. Use a quantitative reference material or test suspension with a known number of Listeria monocytogenes 4b.
    2. Dilute this material so 50 to 120 cfu is inoculated on the ALOA by spreading for example 0.1 mL.
    3. Do this also for the reference media (TSA).
    4. Incubate at (37 ± 1) °C for (48 ± 4) h.
    5. Count the colonies on the medium under test (ALOA) and the reference media (TSA).
    6. Calculate the productivity ratio PR. The criteria is: PR ≥ 0,5.
    7. If the PR exceeds 1,4 identify the reason.
    8. For this the total count of colonies obtained on the culture medium under test (ALOA) is divided by the total count of colonies obtained on the defined reference medium (TSA).
    Calculate the productivity ratio PR as follows: PR  NS
    N0
    Ns = total count test medium
    N0 = total count reference medium

    Control strain WDCM number Ref. media Incubation Criteria Characteristic reaction
    Listeria monocytogenes 4b
    		TSA (37 ± 1) °C for (48 ± 4) h PR ≥ 0,5 Blue green colonies with opaque halo
    Listeria monocytogenes 1/2a
    00109 opt
    The best practice for Quantitative control of Productivity for media control of 'ALOA' is based on the data displayed in this table according ISO 11290-2.

  • Qualitative control of Selectivity

    • Best Practice ALOA

    Qualitative control of Selectivity solid medium

    1. Use a reference material or test suspension with a known number of Escherichia coli and Enterococcus faecalis.
    2. Streak the test strain (Escherichia coli and Enterococcus faecalis) as a single line on the media under test (ALOA).
    3. Multiple strains can be streaked on the plate as long no crossing occurs.
    4. Use a 1 µl loop in case of a concentration of at least 10000 cfu or a 10 µl loop in case of a concentration between 1000 and 10000 cfu.
    5. Incubate at (37 ± 1) °C for (48 ± 4) h.
    6. Asses the amount of growth as, 0 for no growth, 1 for weak growth or 2 for good growth.
    7. For this medium (ALOA) and standard (ISO 11290-2) combination the criteria is: "Total inhibition (0).

    Control strain WDCM number Ref. media Incubation Criteria Characteristic reaction
    Escherichia coli
    or
    		(37 ± 1) °C for (48 ± 4) h Total inhibition (0) No growth
    + Enterococcus faecalis
    or
    The best practice for Qualitative control of Selectivity for media control of 'ALOA' is based on the data displayed in this table according ISO 11290-2.

  • Qualitative control of Specificity

    • Best Practice ALOA

    Qualitative control of Specificity solid medium

    1. Use a reference material or test suspension with a known number of Listeria innocua.
    2. Streak the test controlstrain (Listeria innocua) on the media under test (ALOA) to obtain discrete colonies.
    3. Use a 1 µl loop in case of a concentration of at least 1000 cfu/mL.
    4. Incubate at (37 ± 1) °C for (48 ± 4) h.
    5. Asses the presence, absence and/or grade of expression of biochemical responses and colony sizes and morphology.
    6. For this medium (ALOA) and standard (ISO 11290-2) combination the characteristic reaction is: Blue green colonies without opaque halo.

    Control strain WDCM number Ref. media Incubation Criteria Characteristic reaction
    Listeria innocua
    		(37 ± 1) °C for (48 ± 4) h Growth Blue green colonies without opaque halo
    The best practice for Qualitative control of Specificity for media control of 'ALOA' is based on the data displayed in this table according ISO 11290-2.