Scribd Logo
Upload

Welcome to Scribd!

  • 92 views

    1056 PDF

    Uploaded by

    Vishwas garg
    This document provides instructions for using STANDARD METHODS AGAR (P.C.A) to enumerate bacteria of sanitary significance found in foods and other materials. The agar contains nutrients necessary for bacterial growth including enzymatic digest of casein, yeast extract, glucose, and bacteriological agar. It is recommended by APHA and ISO standards for total microbial plate counts, with incubation times and temperatures varying depending on the standard. The agar supports growth of several test microorganisms including various Escherichia coli and Staphylococcus species.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    1056 PDF

    Uploaded by

    Vishwas garg
    92 views2 pages
    This document provides instructions for using STANDARD METHODS AGAR (P.C.A) to enumerate bacteria of sanitary significance found in foods and other materials. The agar contains nutrients necessary for bacterial growth including enzymatic digest of casein, yeast extract, glucose, and bacteriological agar. It is recommended by APHA and ISO standards for total microbial plate counts, with incubation times and temperatures varying depending on the standard. The agar supports growth of several test microorganisms including various Escherichia coli and Staphylococcus species.

    Original Title

    1056.pdf

    Copyright

    © © All Rights Reserved

    Available Formats

    PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    This document provides instructions for using STANDARD METHODS AGAR (P.C.A) to enumerate bacteria of sanitary significance found in foods and other materials. The agar contains nutrients necessary for bacterial growth including enzymatic digest of casein, yeast extract, glucose, and bacteriological agar. It is recommended by APHA and ISO standards for total microbial plate counts, with incubation times and temperatures varying depending on the standard. The agar supports growth of several test microorganisms including various Escherichia coli and Staphylococcus species.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    92 views2 pages

    1056 PDF

    Uploaded by

    Vishwas garg
    This document provides instructions for using STANDARD METHODS AGAR (P.C.A) to enumerate bacteria of sanitary significance found in foods and other materials. The agar contains nutrients necessary for bacterial growth including enzymatic digest of casein, yeast extract, glucose, and bacteriological agar. It is recommended by APHA and ISO standards for total microbial plate counts, with incubation times and temperatures varying depending on the standard. The agar supports growth of several test microorganisms including various Escherichia coli and Staphylococcus species.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 2

    STANDARD METHODS AGAR (P.C.A) Acc.

    to APHA and ISO 4833

    CAT Nº: 1056 For total microbial plate count in milk and other materials of sanitary significant
    (APHA* formula) and ISO 4833

    FORMULA IN g/l
    Enzymatic Digest of Casein 5.00 Glucose (anhydrous) 1.00

    Yeast Extract 2.50 Bacteriological Agar 15.00

    Final pH 7.0 ± 0.2 at 25ºC

    PREPARATION
    Suspend 23.5 grams of the medium in one liter of distilled water. Mix well and dissolve by heating with frequent
    agitation. Boil for one minute until complete dissolution. Dispense into appropriate containers and sterilize in autoclave at
    121°C for 15 minutes. The prepared medium should be stored from 8-15°C. The color is clear amber, slightly
    opalescent.

    The dehydrated medium should be homogeneous, free-flowing and light toasted in color. If there are any physical
    changes, discard the medium.

    USES
    STANDARD METHODS AGAR (P.C.A.) (PLATE COUNT AGAR) is recommended by APHA when enumerating bacteria of
    sanitary interest, which are indicators of contamination or microbial load in foods.

    Enzymatic Digest of Casein provides nitrogen, vitamins, minerals and amino acids essential for growth. Yeast extract is
    source of vitamins, particularly the B-group. Dextrose is the fermentable carbohydrate providing carbon and energy.
    Bacteriological agar is the solidifying agent.

    In general, 1 ml of the appropriate test dilution is added to the sterile medium at a temperature of 44 – 45°C, mixed
    gently and poured into sterile Petri dishes. Alternatively, dispense a portion of each test dilution (e.g., 0.1, 0.01 ml) into
    separate sterile Petri dishes. Add 10 – 12 ml of tempered (45°C) Standard Methods Agar to Petri dishes containing test
    dilutions. Swirl the dishes to thoroughly mix the medium and test dilution. Allow plates to cool and solidify.

    Incubate the Petri dishes at 32 ± 2°C for 18 – 48 hours and count the developed colonies. Consult the specific texts of
    APHA for the particular sample applications.

    This medium is recommended by the ISO normative 4833 for the colony count technique of microorganisms at 30ºC.
    Inoculate 1ml of the sample, (if necessary 2 continuous decimal dilutions to be able to count between 15-300 colonies
    per plate), put 12-15 ml per plate of agar cooled to 44 - 47ºC in each Petri dish. The time of preparation shouldn’t
    exceed 45 minutes. Invert the plates and incubate at 30 ± 1ºC for 72 ± 3 hours. Post incubation count the colonies.

    MICROBIOLOGICAL TEST
    The following results were obtained in the performance of the medium from type cultures after incubation at a
    temperature of 32 ± 2ºC and observed after 18-48 hours

    Microorganisms Growth
    *Escherichia coli ATCC 25922 Good
    Escherichia coli ATCC 8739 Good

    LABORATORIOS CONDA, S.A. www.condalab.com


    Staphylococcus aureus ATCC 25923 Good
    *Staphylococcus aureus ATCC 6538 Good
    Staphylococcus epidermidis ATCC 12228 Good
    *Bacillus subtilis ATCC 6633 Good
    * According ISO 4833 Incubate at 30ºC for 72 ± 3 hours

    BIBLIOGRAPHY
    International Standard ISO 4833 Microbiology of food and animal feeding stuffs -- Horizontal method for the enumeration of
    microorganisms -- Colony-count technique at 30 ºC
    Standard Methods for the Examination of Dairy Products, 13th Ed. APHA, 1972. American Public Health Association.
    Recommended Methods for the Microbiological Examination of Foods, APHA Inc. New York, 1958. Standard Methods for the
    Examination of Water and Wastewater, APHA Inc. New York, 1960.
    *APHA: American Public Health Association Inc.

    ISO

    STORAGE 25ºC

    Once opened keep powdered medium closed to avoid hydration.


    2ºC

    LABORATORIOS CONDA, S.A. www.condalab.com

    You might also like