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Lab Analyses. what they can tell you what they can’t and how to know the difference. Sample Types. direct microscopic bulk sample, tape lift, BioStrip culture samples bulk sample, swab, Hycon spore traps Air-O-Cell, MCEM, Micro-5 viable air samples RCS, Andersen. Direct Microscopic.

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Lab analyses

Lab Analyses

what they can tell you

what they can’t

and how to know the difference


Sample types
Sample Types

  • direct microscopic

    • bulk sample, tape lift, BioStrip

  • culture samples

    • bulk sample, swab, Hycon

  • spore traps

    • Air-O-Cell, MCEM, Micro-5

  • viable air samples

    • RCS, Andersen


  • Direct microscopic
    Direct Microscopic

    will tell you

    • is there a fungus growing here?

    • what is it?

      but not

    • how much of it is there?

    • how widespread is it?

    • how long has it been there?


    Sample types1
    Sample Types

    • direct microscopic

      • bulk sample, tape lift, BioStrip

  • culture samples

    • bulk sample, swab, Hycon

  • spore traps

    • Air-O-Cell, MCEM, Micro-5

  • viable air samples

    • RCS, Andersen


  • Cultured samples
    Cultured Samples

    can tell you

    • could viable fungal propagules be isolated from the substrate?

      • will they grow and thrive in vitro?

    • what type of fungi were isolated?

    • how many?

      but not

    • were they actually growing there?


    Sample types2
    Sample Types

    • direct microscopic

      • bulk sample, tape lift, BioStrip

  • culture samples

    • bulk sample, swab, Hycon

  • spore traps

    • Air-O-Cell, MCEM, Micro-5

  • viable air samples

    • RCS, Andersen


  • Spore traps
    Spore Traps

    can tell you

    • how many fungal elements were in the air at the moment of sampling?

    • based on their appearance, to which fungal grouping do they belong?


    Spore traps1
    Spore Traps

    can’t tell you

    • are the elements living or dead?

    • precisely what are they?

    • is there fungal growth in the area, and where?

    • will they still be there tomorrow?

    • is the room safe?


    Sample types3
    Sample types

    • direct microscopic

      • bulk sample, tape lift, BioStrip

  • culture samples

    • bulk sample, swab, Hycon

  • spore traps

    • Air-O-Cell, MCEM, Micro-5

  • viable air samples

    • RCS, Andersen


  • Viable air samples
    Viable Air Samples

    can tell you

    • how many culturable fungal elements were in the air at the moment of sampling?

    • in culture, will they produce enough distinguishing characteristics to name them?

    • if so, to which genus and/or species do they belong?


    Viable air samples1
    Viable Air Samples

    can’t tell you

    • is there non-viable fungal material in the air?

    • is there fungal growth in the area, and where?

    • will they still be there tomorrow?

    • is the room safe?


    Direct microscopy
    Direct Microscopy

    SAMPLE NO.:

    1

    2

    3

    4

    5

    6

    Location:

    wine

    cellar

    ballroom

    library

    drawing room

    powder room

    attic

    FUNGAL IDENTIFICATION: ELEMENTS:

    MICROSCOPIC OBSERVATIONS (RATING):

    Acremonium mycelia

    spores

    -

    -

    -

    -

    -

    -

    -

    -

    -

    -

    1+

    2+

    Cladosporium mycelia

    spores

    2+

    3+

    -

    3+

    -

    1+

    -

    tr

    -

    -

    -

    -

    Penicillium mycelia

    spores

    -

    -

    -

    -

    -

    1+

    -

    -

    -

    -

    -

    2+

    Stachybotrys mycelia

    spores

    -

    -

    -

    -

    -

    -

    -

    -

    tr

    1+

    1+

    1+

    basidiospores NOS

    -

    tr

    -

    tr

    -

    -

    hyphal fragments, pigmented

    -

    tr

    -

    tr

    -

    2+

    unknown spores, pigmented, globose

    -

    tr

    -

    -

    -

    tr

    OTHER OBSERVATIONS:

    inorganic and organic microdebris

    1+

    1+

    2+

    3+

    -

    1+

    mites

    -

    -

    -

    -

    -

    tr

    mite faeces

    -

    -

    -

    -

    -

    1+

    FUNGAL GROWTH INDICATED?:

    Y

    Y

    ?

    N

    Y

    Y


    Cultured samples1
    Cultured Samples

    Cultured Bulk Samples

    SAMPLE NO.:

    B1

    B2

    B3

    B4

    B5

    B6

    Medium:

    RBA

    RBA

    RBA

    RBA

    RBA

    RBA

    LOD (CFU/g):

    3100

    240

    480

    110

    4500

    800

    Location:

    carpet

    air filter

    dust

    pipe wrap

    carpet

    fibreglass

    FUNGAL IDENTIFICATION:

    COMPOSITION (CFU/g)*

    Alternaria alternata

    0

    0

    0

    0

    27,000

    0

    Aspergillus versicolor group

    310,000

    1,200

    5,800

    0

    0

    0

    Cladosporium cladosporioides

    6,200

    720

    0

    0

    9,000

    0

    Penicillium subgenus Penicillium

    0

    240

    0

    0

    9,000

    12,000

    sterile mycelium

    0

    480

    8,200

    0

    0

    0

    yeast

    0

    720

    900

    0

    450,000

    0

    TOTAL (CFU/g):

    316,200

    3,360

    14,900

    NFG

    500,000

    12,000


    Microbial populations in duct dust
    Microbial populations in duct dust

    • Standard method to test duct dust for mould is to calculate CFU/g (NADCA)

    • This method does not work

    • Better to test CFU/100cm2

    • Here's why...


    BEFORE CLEANING:

    Mass of dust sampled = 0.1 g

    Yielded 100 fungal colonies

    Total fungal concentration = 1000 CFU/g

    AFTER CLEANING:

    Mass of dust sampled = 0.01 g

    Yielded 50 fungal colonies

    Total fungal concentration = 2000 CFU/g


    Cultured samples2
    Cultured Samples

    Cultured Swab Samples

    †- = not observed; 1+ = 100-101 CFU/plate; 2+ = 101-102 CFU/plate; 3+ = 102-103 CFU/plate; 4+ = >103



    Spore traps2
    Spore Traps

    AIR-O-CELL SAMPLE NO.:

    1

    2

    3

    4

    5

    6

    Location:

    laundry

    utility rm

    kitchen

    bedroom

    attic

    outdoors

    Serial #:

    XXXXXXX

    XXXXXXX

    XXXXXXX

    XXXXXXX

    XXXXXXX

    XXXXXXX

    Expiry date:

    12/08

    12/08

    12/08

    12/08

    12/08

    12/08

    Volume (L):

    75

    75

    75

    75

    75

    75

    Magnification (x):

    500

    500

    500

    1000

    1000

    1000

    Background (rating)§:

    1+

    1+

    1+

    2+

    3+

    2+

    No. of transects enumerated:

    10

    10

    10

    3

    10

    5

    FUNGAL IDENTIFICATION†:

    COMPOSITION (raw count) approx. % of TOTAL‡

    Alternaria

    (2)

    7%

    (tr)

    tr

    (4)

    4%

    (tr)

    tr

    (4)

    6%

    (50)

    31%

    ascospores NOS

    (3)

    11%

    (2)

    4%

    (16)

    16%

    -

    (16)

    25%

    (20)

    12%

    Aspergillus / Penicillium

    -

    -

    -

    (126)

    69%

    (2)

    3%

    (tr)

    tr

    basidiospores NOS

    (12)

    44%

    (10)

    19%

    (62)

    63%

    -

    (22)

    34%

    (62)

    39%

    Cladosporium

    -

    (30)

    58%

    (11)

    11%

    (3)

    tr

    (10)

    16%

    (12)

    7%

    hyphal fragments, pigmented

    (1)

    4%

    (2)

    4%

    (2)

    2%

    -

    (2)

    3%

    (6)

    4%

    Stachybotrys

    (1)

    4%

    -

    -

    -

    -

    -

    unknown spores, hyaline, globose

    (6)

    22%

    (6)

    12%

    (3)

    3%

    (45)

    25%

    (3)

    5%

    (3)

    tr

    unknown spores, pigmented, globose

    (2)

    7%

    (2)

    4%

    (1)

    tr

    (8)

    4%

    (5)

    8%

    (8)

    5%

    SUMMARY DATA¶:

    TOTAL (raw count):

    27

    52

    99

    182

    64

    161

    LOD (elements / m3):

    44

    44

    44

    291

    87

    175

    TOTAL (elements / m3):

    1,200

    2,300

    4,300

    53,000

    5,600

    28,000


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