Milk Safety References
National Conference on Interstate Milk Shipments (NCIMS)

CULTURAL PROCEDURES -
GENERAL REQUIREMENTS
[Unless otherwise stated all tolerances are ±5%]

1. __________ Work Area
   1. __________ Level table or bench, ample working space and utilities
   2. __________ Clean, well ventilated, temperature 16 27C reasonably free from dust and drafts
   3. __________ Well lighted, > 50 foot candles at working surface (pref. 100)
   4. __________ Microbic density of air £ 15 colonies/plate in 15 min exposure, or £ 10 colonies/PAC plate in 15 min exposure, if not corrective actions taken
   5. __________ Freedom from congestion and traffic, only compatible laboratory functions performed
   6. __________ Safe working environment - Refer to OSHA
      1. __________ Eating and drinking not permitted in laboratory
      2. __________ Food and drinks for consumption not stored in laboratory
      3. __________ Analysts wear buttoned/snapped lab coats/uniforms and protective eye-wear, lab coats/uniforms remain on-site
      4. __________ Safety equipment available
      5. __________ MSDS sheets in laboratory available to analysts
      6. __________ Has functioning fume hood with acceptable sash (if necessary, see DMSCC procedure)
      7. __________ Flammable solvent areas continuously well ventilated and temperature controlled
      8. __________ Proper disposal of potentially hazardous materials
         1. __________ Contaminated samples disposed of properly
         2. __________ Contaminated glassware or plasticware disposed of or decontaminated properly
         3. __________ Hazardous chemical disposed of properly
   7. __________ Storage Space
      1. __________ Cabinets, drawers, and shelves adequate
   8. __________ Areas neat, clean and orderly
   9. __________ Floors clean, walls and ceilings in good repair
   10. __________ Laboratory free of insects and rodents
2. __________ Records
   1. __________ All laboratory related records maintained and available for announced surveys
      1. __________ Three (3) years for state central labs
      2. __________ Two (2) years for other labs, minimum requirement, States may require longer periods)
   2. __________ Quality control and sample records available to laboratory evaluation officer during survey
   3. __________ Records contain written corrective actions when taken
   4. __________ Records written in ink or other indelible substance, pencil or erasable ink not allowed
   5. __________ Corrections to quality control records, bench sheets and reports follow the requirements below:
      1. __________ Make a single line through the incorrect information
      2. __________ Write in the correct information next to the incorrect information
      3. __________ Person making the correction initials the information
      4. __________ If not obvious, include reason for correction
   6. __________ Requirements for electronic/computer records
      1. __________ Software must be well documented
      2. __________ Protocols and policies must be clearly documented
      3. __________ Records must be indexed and cross referenced to allow easy review, or must be printed and made available
      4. __________ Records must be secure from unauthorized access and changes
      5. __________ When corrections are necessary the old information must be retained, the person making the correction must be identified and the reason for the change recorded
      6. __________ If records are not available at time of audit, facility will be cited for not having records and will be subject to penalties

   APPARATUS & MATERIALS

3. __________ Thermometers
   1. __________ National Institute of Standards and Testing (NIST) Certified Thermometer, or equivalent, with certificate Serial Number ___________________ ________ ___________________
      1. __________ Graduation interval not more than 0.5C (0-100C) otherwise not more than 1.0C (< 0 or > 100C)
      2. __________ Calibration date on certificate ______________
      3. __________ Annually, checked at the ice point Date _______
   2. __________ Range of test thermometers appropriate for designated use
      1. __________ Mercury-in-glass, alcohol/spirit or digital in degrees centigrade
      2. __________ Plastic lamination recommended for mercury thermometers
   3. __________ raduation interval not more than 0.5C (0-100C) otherwise not more than 1.0C (< 0 or > 100C)
   4. __________ Accuracy of test thermometers checked against certified thermometer
      1. __________ Accurate to ±1C when checked at temperature(s) of use
      2. __________ Results recorded and thermometers tagged
         1. __________ Tag includes identification/location, date of check, calibration temperature and correction factor(s) (read to within ±0.5C)
   5. __________ All test thermometers accuracy checked before initial use and annually, including autoclave maximum registering and hot air oven thermometers
   6. __________ Electronic thermometers checked before initial use and annually as described above
   7. __________ Automatic temperature recording instruments, if used, compared weekly against an accurate thermometer, results recorded
   8. __________ Dial thermometers not used in the laboratory
4. __________ Refrigeration (Sample ___________________________) ________

   __________(Reagent ___________________________)

   1. __________ Size adequate for workload
   2. __________ Maintains samples at 0 4.4C; if temperature out of range, record samples as not analyzed (NA)
   3. __________ Used for storage of milk or milk products, media and reagents only
      1. __________ Not to be used to store food or drink
   4. __________ Record temperature (corrected) daily, in AM and PM, from two thermometers with bulbs immersed in liquid (in sealed containers)
   5. __________ Thermometers located on upper and lower shelves of us
5. __________ Freezer (__________________________)
   1. __________ Size adequate for workload
   2. __________ Maintains -15C or below
   3. __________ Used for storage of frozen milk products, controls, media and reagents only
      1. __________ Not to be used to store food or drink
   4. __________ Record temperature (corrected) daily, in AM and PM, thermometer with bulb immersed in antifreeze liquid (in sealed containers)
6. __________ Pipets (Glass ________ Plastic ________ Pipettor ________)
   1. __________ Appropriate capacity.
   2. __________ Must conform to APHA specifications
   3. __________ Graduations distinctly marked with contrasting color
   4. __________ Discard those with broken tips, scratches or other defects
   5. __________ Pipettors, calibrated, fixed volume or electronic only
      1. __________ Calibrate with ten (10) consecutive weighings once every 6 months (using separate tip for each weighing), average of all 10 weighings must be ±5% of specified delivery volume (by weight, or ³ 1.0 mL by volume using class A graduated cylinder), records maintained
      2. __________ Or, calibrate with 10 consecutive readings once every 6 months using the Artel PCS Pipette Calibration System, average of all 10 readings must be ±5% of specified delivery volume, records/ printouts maintained
         1. __________ Instrument, printer connected by manufacturer's supplied cable or instrument connected to computer via serial cable
         2. __________ Instrument and printer (if applicable) connected to 120v/60Hz power
         3. __________ Reagent kits and Instrument Calibrator kits stored at room temperature

            __________1. Lot # __________ Exp. Date __________

         4. __________ Reagent Blanks and Sample solutions are the same lot
         5. __________ Certificates of Calibration for Reagent Kit and Instrument Calibrator kit maintained in records
         6. __________ Instrument Validation Guide available
         7. __________ PCS Pipette Calibration System Procedure, follow manufacturer's Procedure Guide and instrument prompts
            1. __________ Uncover and insert Blank into the instrument
            2. __________ Determine which volumes are to be calibrated
            3. __________ Select the correct Sample Solution and aliquot sufficient amount into working vessel provided
            4. __________ Using the Pipettor to be verified, aspirate the Sample Solution from the working vessel and deliver it into the Blank seated in the instrument
            5. __________ When appropriate number of data are collected, press 'End of Run' button
            6. __________ Record results and file Pipette Calibration Certificate (printout)
         8. __________ PCS Pipette System Quality Control
            1. __________ Following manufacturer's Procedure Guide and instrument prompts, perform an instrument calibration every 30 days or just prior to use
            2. __________ Record results and file Calibration Certificate (printout)
         9. __________ PCS Calibration System Validation
            1. __________ Upon receipt, validate the instrument by following the manufacturer's protocol
      3. __________ Pipettors etched with identification (imprinted serial numbers acceptable) and tagged with date calibrated
      4. __________ Tips (sterile for plate counts) appropriate to pipettor(s) being used
7. __________ Pipet Containers
   1. __________ Used for sterilization, storage; non-toxic
8. __________ Dilution Bottles and Closures, reusable
   1. __________ Bottles of borosilicate glass __________ or approved plastic __________ with smooth tops
   2. __________ Capacity 150 mL, indelibly marked at 99±1 mL level
   3. __________ Closure non toxic rubber stopper or plastic screw cap with liner
   4. __________ New Bakelite type plastic caps and closures treated to remove toxic residues, tested using a B. stearothermophilus type assay
   5. __________ Discard bottles and caps with chips, cracks, scratches or other defects
9. __________ Petri Dishes (Glass ______ or Plastic ______)
   1. __________ Bottom at least 80 mm I.D., and 12 mm deep for plate counts

      __________Brand __________________

   2. __________ Scanner (_____________________)

      __________Brand __________________

   3. __________ Bottom flat and free from bubbles, scratches, or other defects
10. __________ Petri Dish Container
    1. __________ Used for sterilization, storage; non toxic
11. __________ Hot-Air Sterilizing Oven (________________________)
    1. __________ Sufficient size to prevent crowding of interior in normal usage
    2. __________ Constructed to provide uniform temperature in chamber
    3. __________ Thermometer or temperature recorder with adequate range (to 220C)
       1. __________ Thermometer checked at temperature of use for accuracy before initial use, records maintained
       2. __________ Thermometer bulb immersed in sand
    4. __________ Records maintained for each sterilization cycle including date, start up time, time sterilization temperature reached, and length of time at sterilization temperature
    5. __________ Temperature indicator used each load
    6. __________ Performance checked with full load and recorded quarterly (preferably weekly) using spore (B. subtilus) strips, include positive control check, results maintained
       1. __________ Brand: _____________________________
       2. __________ Lot #: __________ Exp. Date: _____
12. __________ Sterilization by Dry Heat
    1. __________ Material in center of load heated to ³ 170C for ³ 2 hrs
    2. __________ Oven not crowded (< 75% of shelf in gravity type, 90% in forced air type)
13. __________ Autoclave (Media _______________________________)

    __________ (Waste _______________________________)

    1. __________ Sufficient size to prevent crowding of chamber
    2. __________ Thermometer or temperature recorder controller properly located to register chamber temperature
    3. __________ Has pressure gauge and properly adjusted safety valve
    4. __________ Connected to suitable saturated steam line or steam generator
    5. __________ Chamber temperature checked at least quarterly (preferably more frequently, ex. weekly with sterility check) with full load with maximum registering thermometer and results recorded
    6. __________ Cycle timing checked quarterly and found to be accurate, record maintained
    7. __________ Records maintained for each sterilization cycle including date, start up time, temperature and time temperature reached, length of time at temperature, time at end of run, time removed and item(s) autoclaved (including waste)
       1. __________ Strip recorders that provide the above information are acceptable if strips (or copies) are maintained in permanent record, include items autoclaved, time removed and initials
       2. __________ Circular charts must be interpreted and must have written records to verify the information stated above
    8. __________ Temperature indicator used each load
    9. __________ Performance checked with full load and results recorded weekly using spore (B. stearothermophilus) strips or suspensions, include positive control check, results maintained
       1. __________ Brand: _____________________________
       2. __________ Lot #: __________ Exp. Date: _____
    10. __________ Routine maintenance performed and records maintained
14. __________ Sterilization by Moist Heat
    1. __________ Media autoclaved at 120±1C
       1. __________ Dilution buffer blanks for 15 min (30 min optional)
       2. __________ Media for 15 min (sugar broths as per manufacturer instructions)
    2. __________ Media autoclaved within 1 hr of preparation
    3. __________ Dilution buffer autoclaved on same day prepared
    4. __________ Stoppers or caps slightly loosened to permit passage of steam and air
    5. __________ All air expelled from autoclave before pressure allowed to rise
    6. __________ Autoclave will reach 120±1C within 15 min (5 min pref) of starting air-exhaust
    7. __________ Properly operating and calibrated temperature gauge (not a pressure gauge) relied on to insure sterilization
    8. __________ After sterilization, pressure gradually reduced (³ 15 min) and media removed promptly when atmospheric pressure is reached
    9. __________ Total time in autoclave less than 1 hour
15. __________ Incubator and/or Incubator Room (SPC, PAC and Coliform)

    __________(#1 ______________________________)

    __________(#2 ______________________________)

    1. __________ Sufficient size to prevent crowding of interior
    2. __________ Shelves placed to assure uniformity at 32C±1C
    3. __________ Chamber temperatures measured by not less than two thermometers with bulbs immersed in liquid (in sealed containers)
    4. __________ Thermometer located on the top and bottom shelves of use
    5. __________ Temperature (corrected) recorded from each thermometer twice daily (AM and PM)
    6. __________ Agar (10 - 12 mL) in SPC plates and/or (1 mL) in PAC plates must not lose more than 15% weight after 48 hrs incubation
       1. __________ Agar weight loss of SPC and/or PAC plates tested quarterly and results recorded
          1. __________ Test minimum of two (2) plates/films per shelf in use, one on each side of shelf, preferably test 10 plates evenly distributed throughout the incubator
       2. __________ Corrective action taken when criteria not met and records of corrective actions maintained
          1. __________ If weight loss is out of compliance take corrective actions (humidify incubator, reduce air flow, etc.) and retest as above and record
          2. __________ Use more agar (15 - 20 mL), if this option used laboratory must document that this amount of of agar is routinely used for plating
16. __________ Colony Counter
    1. __________ Quebec dark field model or equivalent with satisfactory grid plate
17. __________ Hand Tally, accurate
18. __________ pH Meter (Milk Lab _________________________)

    __________ (Media Prep _________________________)

    1. __________ Electronic only, readable to 0.1 pH units
    2. __________ Daily calibration and slope records and maintenance log maintained when in use
    3. __________ Record date electrodes (double junction reference pref) put into service (write in QC record and tag probe)
19. __________ pH Measurement
    1. __________ All measurements made at room temperature
    2. __________ Instrument standardized with known buffer solutions
       1. __________ Three commercially prepared standard solutions used
       2. __________ Each aliquot used once and discarded
       3. __________ pH 4, 7 and 10 suggested for linearity and proper function of meter
       4. __________ Slope determined (95 - 102%) ____________ each time meter calibrated, records maintained
    3. __________ Medium pH recorded each time measured
    4. __________ Final (after sterilization) pH of each batch of medium determined before use, records maintained
       1. __________ Standard Methods Agar, pH 7.0±0.2
       2. __________ Violet Red Bile Agar, pH 7.4±0.2
       3. __________ Brilliant Green Bile Broth, pH 7.2±0.2
       4. __________ PM Indicator Agar, pH 7.8±0.2
       5. __________ Buffered rinse solution, 7.2±0.2
       6. __________ Nutrient broth, pH 6.8±0.2
       7. __________ Letheen Broth, pH 7.0±0.2
       8. __________ Lauryl Tryptose Broth (LST), pH 6.8±0.2
       9. __________ M-Endo Agar or Broth, pH 7.2±0.2
       10. __________ MMO-MUG Medium, pH 7.4±0.2
       11. __________ Stock phosphate buffer, pH 7.2±0.2
       12. __________ Dilution buffer, pH 7.2±0.2
20. __________ Balance (Milk _____________________________)

    __________(Media _____________________________)

    __________(Analytical _____________________________)

    1. __________ Electronic only, sensitive to ³ 0.1g for general laboratory purposes and proper sensitivity for calibrations and antibiotics
    2. __________ Class S or S1, or equivalent ASTM 1, 2, or 3, weights
       1. __________ Certificate or other verification of authenticity
       2. __________ Free from excessive wear, filth and corrosion
       3. __________ Weights within class tolerance
    3. __________ Checked monthly with weights corresponding to normal use of balance (ex. 100, 200, 500, 1000 mg, etc. for analytical balances, and 5, 10, 25, 50, 100, 150g, etc. for other balances), records maintained
    4. __________ Checked at least annually, or when weights out of tolerance, by a qualified representative for good working order with proof of check in laboratory
       1. __________ Date of last check __________
21. __________ Water Baths
    1. __________ Thermostatically controlled to appropriate temperature(s)
    2. __________ Water circulation capability, baths up to 64C
    3. __________ Appropriate size for work loads
    4. __________ Suitable water level maintained
22. __________ Mechanical Dilution Bottle Shaker
    1. __________ Type described in SMEDP, 11th Edition
    2. __________ Other acceptable ____________________
23. __________ Microwave Oven for Melting Media
    1. __________ Analysts instructed to take extreme caution as media expands rapidly at the boiling point
24. __________ Microbiologically Suitable (MS) Water
    1. __________ Type _________________________________
    2. __________ System used _______________________________
    3. __________ Monthly testing criteria
       1. __________ Standard plate count < 1,000 colonies/mL(< 10,000 colonies/mL if stored)
       2. __________ Total chlorine residual negative, recorded as less than the detection limit of test used
       3. __________ Resistance exceeds 0.5 megohm/cm or conductance is less than 2.0 umhos/cm (at 25C)
          1. __________ Brand: _______________ Std. ____________
          2. __________ Test performed in another lab
    4. __________ Tested annually for total metals (Pb, Cd, Cr, Cu, Ni and Zn), not to exceed 0.05 mg/L for each metal and not to exceed 0.1 mg/L total for all metals
    5. __________ If criteria not met, corrective action(s) taken and recorded in QC record
    6. __________ Records maintained
25. __________ Dilution Buffer and Blanks
    1. __________ Stock phosphate buffer (Date prepared ______________)
       1. Prepared in laboratory (34g KH₂PO₄/liter) with MS water
       2. __________ Purchased commercially prepared _______________
       3. __________ Lot No. __________ Exp. Date __________

          __________ Rcd. Date __________ Date Opened __________

       4. __________ Place in small containers (£ 100 mL), autoclave and store in refrigerator
    2. __________ Stock MgCl2 Solution, Optional (Date prepared ___________)
       1. __________ Prepared in laboratory (38g MgCl₂/liter or 81.1 g MgC1₂·6H₂0/liter) with MS water
       2. __________ Purchased commercially prepared ________________
       3. __________ Lot No. __________ Exp Date __________

          __________ Rcd. Date __________ Date Opened __________

       4. __________ Place in small containers (£ 100 mL), autoclave and store in refrigerator
    3. __________ Prepare dilution buffer with 1.25 mL stock buffer/liter of MS water
       1. __________ Optionally, add 5 mL of stock MgCl₂/liter of MS water
    4. __________ Dilution bottles filled to contain 99±2 mL dilution buffer after sterilization
       1. __________ After sterilization and after cool visually observe and discard any blanks with < 97 or > 101 mL
       2. __________ Of remaining blanks appearing to have the correct volume, check 1 blank for every 25 that were made using a class A graduate cylinder (or equivalent)
       3. __________ Maintain records of volume checks, including batch size
       4. __________ If any blanks out of tolerance, discard entire lot, record lot as discarded
    5. __________ Blanks tested at 6 month intervals for toxic substances
       1. __________ Plate milk dilution at 0, 15, 30, 45 min
       2. __________ If the 45 min count is 20% less than 0 min count, determine cause and retest after correction made, records maintained
    6. __________ Store

       __________ Lot No. __________ Exp. date __________

       __________ Rcd. Date __________

       1. __________ Volume records maintained as above
       2. __________ Toxicity checked as above on each new lot received
       3. __________ Check pH and record
    7. __________ Records maintained
    8. __________ Corrective action taken when criteria not met, records maintained
26. __________ Reagent Chemicals of ACS Grade
27. __________ Media
    1. __________ Use dehydrated medium of correct composition
       1. __________ Each bottle dated on receipt (in lab or by central receiving, which ever first) and when first opened for use
       2. __________ Stored as specified by manufacturer; after opening, each bottle tightly capped following each use
       3. __________ Commercially sealed medium kept no longer than manufacturer's expiration date
       4. __________ Opened bottles used until manufacturer's expiration date
       5. __________ Discarded if any change is noted in appearance or hydration regardless of manufacturer's expiration date
    2. __________ Plate Count Agar _______________
       1. __________ Composition Pancreatic Digest of Casein 5 g
          Yeast Extract 2.5 g
          Glucose 1 g
          Agar 15 g
          MS water to make 1 L
       2. __________ Lot No. _________ Exp. Date __________
          Rcd. Date __________ Date Opened __________
    3. __________ Petrifilm Aerobic Count (PAC) Plate
       1. __________ Lot No. _________ Exp. Date __________
          Rcd. Date __________ Date Opened __________
    4. __________ Violet Red Bile Agar _______________
       1. __________ Composition
          Yeast Extract 3 g
          Peptone or Gelysate 7 g
          Bile Salts 1.5 g
          Lactose 10 g
          Sodium Chloride 5 g
          Neutral Red 0.03 g
          Crystal Violet 0.002 g
          Agar 15 g
          MS water to make
       2. __________ Boil 2 min, temper and use within 3 hours (do not autoclave)
       3. __________ Lot No. _________ Exp. Date __________
          Rcd. Date __________ Date Opened __________
    5. __________ Petrifilm Coliform Count (PCC) Plate
       1. __________ Lot No. _________ Exp. Date __________
          Rcd. Date __________ Date Opened __________
    6. __________ Petrifilm High Sensitivity Coliform Count (HSCC) Plate
       1. __________ Lot No. _________ Exp. Date __________
          Rcd. Date __________ Date Opened __________
    7. __________ Brilliant Green Lactose Bile Broth _______________
       1. __________ Composition
          Peptone or Gelysate 10 g
          Lactose 10 g
          Oxgall 20 g
          Brilliant Green 0.0133 g
          MS water to make 1 L
       2. __________ Lot No. _________ Exp. Date __________
          Rcd. Date __________ Date Opened __________
    8. __________ PM Indicator Agar _______________
       1. __________ Composition
          Beef Extract 3 g
          Peptone 5 g
          Tryptone 1.7 g
          Soytone 0.3 g
          Dextrose 5.25 g
          Sodium Chloride 0.5 g
          Dipotassium Phosphate 0.25 g
          Polysorbate 80 1 g
          Brom Cresol Purple 0.06 g
          Agar 15 g
          MS water to make 1 L
       2. __________ Lot No. _________ Exp. Date __________
          Rcd. Date __________ Date Opened __________
    9. __________ Buffered Rinse Solution _______________
       1. __________ Composition
          Stock Phosphate Buffer 1.25 mL
          10% Na Thiosulfate Solution 5 mL
          Azolectin 4 g
          Tween 20 10 g
          MS water to make 1 L
       2. __________ Weigh hygroscopic Azolectin rapidly and dissolve by heating over boiling water
       3. __________Date prepared
    10. __________ Nutrient Broth (laboratory use only) _______________
        
        1. __________ Composition
           Beef Extract 3 g
           Peptone 5 g
           MS water to make 1 L
        2. __________ Lot No. _________ Exp. Date __________
           Rcd. Date __________ Date Opened __________
    11. __________ Letheen Broth _______________
        (For use with Petrifilm, Do not use diluents containing thiosulfate or sodium citrate)
        1. __________ Composition
           Peptamin 10 g
           Beef Extract 5 g
           Lecithin 0.5 g
           Sorbitan Monooleate 5 g
           Sodium Chloride 5 g
           MS water to make 1 L
        2. __________ Lot No. _________ Exp. Date __________
           Rcd. Date __________ Date Opened __________
    12. __________ Lauryl Tryptose Broth (LST) ________________
        1. __________ Composition
           Tryptose 20 g
           Lactose 5 g
           Dipotassium Phosphate 2.75 g
           Monopotassium Phosphate 2.75 g
           Sodium Chloride 5 g
           Sodium Lauryl Sulfate 0.1 g
           MS water to make 1 L
        2. __________ Lot No. _________ Exp. Date __________
           Rcd. Date __________ Date Opened __________
    13. __________ M-Endo Agar _______________
        1. __________ Composition
           Yeast Extract 1.2 g
           Casitone 3.7 g
           Thiopeptone 3.7 g
           Tryptose 7.5 g
           Lactose 9.4 g
           Dipotassium Phosphate 3.3 g
           Monopotassium Phosphate 1 g
           Sodium Chloride 3.7 g
           Sodium Desoxycholate 0.1 g
           Sodium Lauryl Sulfate 0.05 g
           Sodium Sulfite 1.6 g
           Basic Fuchsin 0.8 g
           Agar 15 g
           MS water to make 1 L
        2. __________ Lot No. _________ Exp. Date __________
           Rcd. Date __________ Date Opened __________
    14. __________ M Endo Broth _______________
        1. __________ Composition
           Yeast Extract 1.5 g
           Casitone 5 g
           Thiopeptone 5 g
           Tryptose 10 g
           Lactose 12.5 g
           Dipotassium Phosphate 4.375 g
           Monopotassium Phosphate 1.375 g
           Sodium Chloride 5 g
           Sodium Desoxycholate 0.1 g
           Sodium Lauryl Sulfate 0.05 g
           Sodium Sulfite 2.1 g
           Basic Fuchsin 1.05 g
           MS water to make 1 L
        2. __________ Lot No. _________ Exp. Date __________
           Rcd. Date __________ Date Opened __________
    15. __________ MMO-MUG Medium _______________
        1. __________ Commercial or lab prepared media containing MMO-MUG
        2. __________ Composition
           Ammonium Sulfate 5 g
           Manganese Sulfate 0.0005 g
           Zinc Sulfate 0.0005 g
           Magnesium Sulfate 0.1 g
           Sodium Chloride 10 g
           Calcium Chloride 0.05 g
           Sodium Sulfite 0.04 g
           Amphotercin B 0.001 g
           o-nitrophenyl-ß-D- galactopyranoside 0.5 g
           4-methylumbellifery-ß-D- glucuronide 0.075 g
           Solanium 0.5 g
           Hepes Buffer
           Sodium Salt 5.3 g
           Organic Acid 6.9 g
           MS water to make 1 L
        3. __________ Lot No. _________ Exp. Date __________
           Rcd. Date __________ Date Opened __________
    16. __________ Charm E*Colite
        1. __________ Lot No. _________ Exp. Date __________
           Rcd. Date __________
28. __________ Medium Preparation
    1. __________ Media making utensils borosilicate glass, stainless steel, or other non-corrosive equipment
    2. __________ Weigh required amount of dehydrated medium or ingredients
    3. __________ Combined with required amount MS water, dissolved and mixed in a suitable container
    4. __________ pH adjusted if necessary
    5. __________ Heated (covered), not under pressure, if necessary, to complete solution (microwave preparation not allowed)
    6. __________ Water restored, as necessary, to compensate for loss due to evaporation
    7. __________ Distributed into suitable containers so that no part of medium is more than 2.5 cm from any surface
       1. __________ In general, containers filled no more than half of total volume .
    8. __________ Suitable container closures used and autoclaved as necessary
29. __________ Prepared Media Storage
    1. __________ Protected from water loss and light
    2. __________ Only screw capped containers kept no more than 6 months
    3. __________ Prepared Charm PMI plates, kept no more than 5 days in sealed container at 0 4.4C (tag with date of preparation)
    4. __________ BGB broth at room temperature
       1. __________ Screw capped tubes for 3 months
       2. __________ Loose (slip) capped tubes for 1 week
       3. __________ Stored in dark
    5. __________ Petrifilm plate storage
       1. __________ Refrigerate unopened packages of Petrifilm plates at or below 8C, if frozen allow 30 min room temperature thaw time before opening packages
       2. __________ Use before expiration date on package
       3. __________ After opening, return unused plates to foil pouch, seal pouch by folding and taping/clipping open end shut
       4. __________ Store re-sealed packages £ 21C, £ 50% relative humidity. Do not refrigerate opened packages.
       5. __________ Use Petrifilm plates within one month after opening package (tag with date opened
    6. __________ Pre-dispensed rinse solutions for containers
       1. __________ Dispense in appropriate volume (20, 50, 100 mL, or other) and sterilize
       2. __________ Perform quality control checks for volume (100±2 mL) as described in cultural procedures item 25d
30. __________ Detergent Suitability Test
    1. __________ Detergent residue test performed if laboratory washes and re-uses glassware (not required if only disposable items used)
    2. __________ Detergent is suitable for laboratory use

       __________ Brand ____________________ Brand ____________________

    3. __________ Test each new brand/lot, records maintained
31. __________ Cleaning Pipets
    1. __________ Used pipets discarded in disinfectant
    2. __________ Rinsed in tap water at 15-30C
    3. __________ Thoroughly washed with suitable detergent and rinsed
    4. __________ Cleaned with strong cleaning solution such as acid dairy cleaner as necessary
    5. __________ Final rinse with MS water
    6. __________ Several pieces from each batch tested (preferably while still wet) for residual acid or alkali with aqueous 0.4% bromthymol blue. If color reaction not dark green to light blue, re-rinse and test again. Records maintained
32. __________ Cleaning Other Glassware and Apparatus
    1. __________ Heated to 85C or disinfected unless pathogens suspected; then sterilization required prior to washing
    2. __________ Washed with hot water and suitable detergent and rinsed
    3. __________ Machine washed (__________________________)
    4. __________ Hand washed ______
    5. __________ Final rinse with MS water
    6. __________ Several pieces from each batch tested (preferably while still wet) for residual acid or alkali with aqueous 0.04% bromthymol blue. If color reaction not dark green to light blue, re-rinse and test again. Records maintained

    SAMPLES

33. __________ Laboratory Requirements
    1. __________ Record time, date, and temperature of samples as received
    2. __________ Determine sample temperature
       1. __________ Insert a pre-cooled thermometer into temperature control (pre-cooling of electronic/digital thermometer probes is not necessary)
       2. __________ Temperature control must be at least half the size of the largest test container
       3. __________ Performed by trained personnel, not by collector, establish record to indicate training performed
    3. __________ Do not accept samples if temperature control for each tank truck and each plant or delivery truck group of samples is missing
    4. __________ Do not accept or test samples if sample containers are leaking
    5. __________ Do not accept or test samples if samples are unprotected and/or submerged in ice/ice water slush
    6. __________ Do not accept fluid samples, which are frozen, for microbial or somatic cell analysis
    7. __________ Do not accept raw samples if sample containers have no head space (about ¾ full)
    8. __________ If milk sample temperature control exceeds 4.4C on receipt, do not test microbiologically (samples may be tested if temperature does not exceed 7C and time of receipt is £ 3 hours from collection and sample receipt temperature is no greater than that at collection)
    9. __________ Store fluid samples at 0 4.4C until tested, if storage temperature exceeds 4.4C prior to testing record as LA
    10. __________ Do not proceed with analysis if above criteria have not been met
34. __________ Sample Bench Sheet Requirements
    1. __________ Must show date, time and temperature collected, along with name of official sampler
    2. __________ Must show date, time and temperature when brought to the laboratory, along with whom received the samples
    3. __________ Must show date and time of analysis, temperature of samples at start of analysis, and names of analyst(s) performing test(s)
    4. __________ Sample bench sheets or records must contain all results (raw and calculated in their proper format) of tests performed and the results of all controls that apply to each test
       1. __________ Plate count procedure controls include:
          1. __________ Microbic air density
          2. __________ Dilution buffer
          3. __________ Pipets
          4. __________ Agar
          5. __________ Temperature of agar at plating (45±1C)
       2. __________ Results of inhibitor test(s) accompany all plate count and coliform results
       3. __________ Control results recorded for each inhibitor test performed
       4. __________ All above recorded on sample bench sheets

    MISCELLANEOUS

35. __________ Laboratory Practices
    1. __________ Personnel adequately trained and/or supervised
    2. __________ Satisfactory participation in annual split samples
    3. __________ Copies of current, applicable FDA 2400 series survey forms in laboratory
    4. __________ Copy of current edition Standard Methods in laboratory
    5. __________ Copy of AOAC Manual of Methods in laboratory if necessary
    6. __________ Copy of written Quality Assurance Plan, required for state central laboratories
    7. __________ Laboratory management has signed and returned the agreement to abide by the provisions of the NCIMS and the procedures for the Evaluation of Milk Laboratories
    8. __________ Laboratory evaluation officer conducted survey unobstructed by laboratory or facility personnel