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While performing additional validation of the PDATR procedure, we found that when dry foods such as spices were filtered, mold colonies did not always develop within 48 h at
25C; rather, reincubation of the filter for an additional 24 h at
25C was often required to obtain results that correlated to the
5-day PDA pour plate method. However, reincubating also resulted in overgrowth of the filter by more rapidly growing components of the flora. Therefore, we designed a new culture medium (YM-11 agar) to improve the development of the slower
growing organisms so that the incubation period could be
maintained at 48 h.
A comprehensive precollaborative study of the 2-day hydrophobic grid membrane filter method incorporating YM-11 agar
was performed with both pure cultures and 26 food product
categories. The new method performed equivalently to the 5day PDA pour plate method (3). After successful completion of
the precollaborative study, we conducted a collaborative study,
the results of which are reported here.
Collaborative Study
A. Principle
Method uses membrane filter imprinted with hydrophobic
material in grid pattern. Hydrophobic lines act as barriers to
pain powder with 100 mL sterile H2O. Unused portion of solution can be stored frozen up to 3 months.
(l) Hemicellulase stock solution.1500 HCU (viscometric
hemicellulase unit)/g. Reconstitute 20.0 g presterilized
hemicellulase powder with 100 mL sterile H2O. Unused portion of solution can be stored frozen up to 3 months.
Items (a), (b), (i), (j), (k), and (l) are available from QA Life
Sciences, Inc., 6645 Nancy Ridge Dr., San Diego, CA 92121.
37C. Correct for additional dilution by filtering 1.2 mL enzyme-treated sample homogenate.
(e) Other foods.Prepare 1:10 dilution by aseptically
weighing 10 g sample into sterile blender jar. Add 90 mL peptone, B(g), or PT diluent, B(h) (see Table 995.21B), and blend
2 min at low speed (10 00012 000 rpm). If enzyme treatment
is needed, combine 5 mL sample homogenate (1:10 dilution)
with 1 mL appropriate enzyme stock solution, mix well, and
incubate 2030 min in H2O bath at 3537C. Correct for additional dilution by filtering 1.2 mL enzyme-treated sample homogenate.
D. Analysis
See Figures 986.32A and 986.32B for filtration unit and filtration unit clamp.
Turn on vacuum source. Place sterile filtration unit on manifold or vacuum flask. Open clamp A. Rotate back funnel portion C. Aseptically place sterile filter, B(a), on surface of
base D. Rotate funnel forward. Clamp shut by sliding jaws L of
stainless steel clamp over entire length of flanges B that extend
from both sides of funnel C and base D and rotating moving
arm K into horizontal (locked) position.
Aseptically add ca 1520 mL sterile H2O to funnel. Pipet
1.0 mL sample homogenate (1:10 dilution) or appropriate volume of enzyme-treated sample homogenate into funnel. Apply
free end of vacuum tubing E to suction hole F to draw liquid
through prefilter mesh G. Aseptically add additional 1015 mL
sterile H2O to funnel and draw through mesh as before. Close
clamp A to direct vacuum to base of filtration unit and draw
liquid through filter.
Rotate moving arm K of stainless steel clamp into unlocked
(ca 45 angle) position and slide jaws L off flanges B. Rotate
back funnel C. Open clamp A.
Place filter on surface of predried YM-11 plate, B(i). Avoid
trapping air bubbles between filter and agar. Incubate plates
50 2 h at 25 1C.
Count all squares containing one or more colonies (positive
squares). Colonies are usually some shade of blue. Examine
filter with illuminated magnification, as some colonies may be
only pinpoint in size. To confirm that filter does not contain any
positive squares, hold up Petri dish and examine horizon of
filter for raised bumps or areas that reflect light differently.
These may be either very small or very pale colonies. Although
hydrophobic lines act as barriers to spread of colonies, some
fast-growing molds produce too much mycelium to remain
completely confined within a single square. If sample contains
fast-growing molds, some colonies may cover more than one
square (spreaders). In case of spreader, count only the square of
origin, which usually has the densest or tallest growth. Do not
count each individual square into which colony has spread.
Count squares containing one or more colonies as described
above. Convert total number of positive squares to MPN index
as follows:
MPN = 1600 loge
1600
1600 x
Garlic Powder
The collaborative study results for garlic powder are presented in Table 1. Collaborator 11 was a statistical outlier, having reported results for all 6 garlic powder samples 23 orders
of magnitude higher than any other collaborator for both the
test and reference methods. Data from this collaborator were
excluded from statistical analysis.
Contamination levels were very low in all 3 independent
lots of this food and consisted virtually entirely of molds.
Counts obtained by the test and reference methods did not differ significantly from each other (Pr > F of 0.4610), and the
repeatability and reproducibility results were similar by both
methods (Table 995.21A).
Walnut Pieces
The collaborative study results for walnut pieces are presented in Table 3. The following data were excluded from statistical analysis: Collaborators 1 and 5 were found to be statistical outliers for the reference method by the rank score test (5),
and collaborators 12 and 14 were statistical outliers for the test
method.
The flora in these samples consisted almost entirely of
molds. The test and reference methods produced results which,
although numerically not very different (differences between
the log10 means were 0.15 or less for each of the 3 lots), were
significantly different based on ANOVA (Pr > F of 0.0001).
Repeatability and reproducibility values were similar for both
methods (Table 995.21A)
Flour/Meal
The collaborative study results for flour/meal are presented
in Table 4. Flora in these samples consisted principally of
molds. There were no significant method deviations or statistical outliers associated with this food product. Differences
between the test and reference methods were not statistically
significant (Pr > F of 0.2905), and repeatability and reproducibility results were similar for both methods (Table 995.21A).
Orange Juice
The collaborative study results for orange juice are presented in Table 5. The following data were excluded from statistical analysis: Collaborator 1 set up the orange juice samples
approximately 1 week late. Data from this collaborator were
found to be statistical outliers for both methods by the rank
score test (5). Collaborator 2 was a statistical outlier for the orange juice for both the test and reference methods, and collaborator 4 was a statistical outlier for the orange juice for the test
method, on the basis of the rank score test (5). Collaborator 14
reported a significant technique error for the reference method
(see description of the error in Raw Ground Beef).
The majority of the flora in these samples consisted of
yeasts, although some molds were also present. The test
method produced results that were significantly higher than the
reference method, based on ANOVA (Pr > F of 0.0001). Results from 1 lot were similar by both methods, but the test
method produced counts more than one-half of a log10 cycle
higher than the reference method for the other 2 lots. Repeatability and reproducibility values were in a similar range for
both methods (Table 995.21A).
Yogurt
The collaborative study results for yogurt are presented in
Table 6. The natural flora in all 3 lots of yogurt multiplied far
more rapidly in the product than expected between the time of
prescreening in the coordinating laboratory and the day specified for initiation of analyses. As a result, many collaborators
reported indeterminately high counts for the test method, which
had an absolute upper counting limit of 107/g, based on the
dilution specified for analysis by the Associate Referee,
whereas the reference method was often able to yield estimated
counts beyond 107/g. Collaborator 1 set up the yogurt samples
approximately 1 week late, and data from this collaborator
were excluded from statistical analysis. Collaborator 14 reported a significant technique error with the reference method
(see description of the error in Raw Ground Beef), and data
from this collaborator were also excluded from statistical
analysis.
Results from 2 of the 3 independent lots of yogurt were numerically higher by the reference method, and results from the
third lot were numerically higher by the test method. The largest difference between the method log10 means for any lot was
0.20. Although the numerical differences were not large, the
methods differed significantly, on the basis of ANOVA (Pr > F
of 0.0465). Repeatability and reproducibility values were in a
similar range for both methods (Table 995.21A).
Yogurt was one of 26 products included in the precollaborative study (3), and differences between the test and reference
methods were not significant for the precollaborative study
samples. Other dairy products included in the precollaborative
study were sour cream, cream cheese, and grated Parmesan
cheese. In all of these cases, differences between the methods
were not statistically significant.
Table 995.21A. Method performance for enumeration of yeast and mold in foods by ISO-GRID and reference
methods
Food
Garlic powder
Lot
Method
Log10 mean,
count/g or
mL
ISO-GRID
Reference
1.23
1.08
0.95
0.90
1.12
1.23
0.34
0.32
0.40
0.44
27.49
29.12
32.09
40.19
0.76
1.01
0.27
0.36
22.35
30.03
1.20
1.01
0.35
0.19
0.43
0.36
28.95
16.11
36.10
30.42
Reference
1.20
1.34
1.48
0.48
0.53
40.00
44.18
ISO-GRID
Reference
1.20
1.18
ISO-GRID
3.61
0.76
1.06
0.27
0.38
7.50
10.60
Reference
ISO-GRID
3.59
3.91
0.81
0.95
1.06
1.32
0.29
0.34
0.38
0.47
8.23
8.70
10.61
11.99
Reference
3.84
0.64
1.01
0.23
0.36
5.94
9.36
ISO-GRID
Reference
4.43
4.28
1.09
0.81
1.68
1.46
0.39
0.29
0.60
0.52
8.82
6.88
13.60
12.03
Overall
ISO-GRID
3.99
Reference
ISO-GRID
3.90
3.57
0.67
0.73
0.24
0.26
6.77
7.15
Reference
3.68
0.53
0.53
0.19
0.19
4.93
5.07
ISO-GRID
Reference
4.08
4.23
0.28
0.22
0.64
0.45
0.10
0.08
0.23
0.16
2.47
1.90
5.59
3.81
ISO-GRID
3.66
0.62
0.70
0.22
0.25
5.97
6.74
3.79
3.76
0.59
0.59
0.21
0.21
5.43
5.43
Overallb
Reference
ISO-GRID
Reference
3.90
ISO-GRID
Reference
3.36
3.52
1.20
1.01
1.20
1.01
0.43
0.36
0.43
0.36
12.80
10.23
12.80
10.23
ISO-GRID
4.63
0.59
1.18
0.21
0.42
4.45
9.05
Reference
4.76
0.76
0.92
0.27
0.33
5.66
6.97
ISO-GRID
Reference
3.38
3.20
1.01
0.73
1.18
1.09
0.36
0.26
0.42
0.39
10.57
8.19
12.46
12.23
Overall
ISO-GRID
3.80
Reference
ISO-GRID
3.83
3.11
0.53
0.64
0.19
0.23
5.91
7.19
Reference
3.32
0.42
0.64
0.15
0.23
4.39
6.85
ISO-GRID
Reference
7.20
6.66
1.04
1.12
1.18
2.10
0.37
0.40
0.42
0.75
5.16
6.06
5.87
11.31
ISO-GRID
6.00
0.73
1.74
0.26
0.62
4.39
10.26
5.45
5.45
1.29
1.54
0.46
0.55
8.35
10.08
Overallb
Reference
ISO-GRID
Reference
5.15
ISO-GRID
Reference
6.54
6.75
0.48
0.31
1.37
1.82
0.17
0.11
0.49
0.65
2.51
1.59
7.44
9.58
A
B
ISO-GRID
6.46
0.45
1.85
0.16
0.66
2.46
10.25
Reference
ISO-GRID
6.61
6.72
0.34
0.59
2.38
1.48
0.12
0.21
0.85
0.53
1.81
3.07
12.85
7.83
Reference
6.66
0.62
1.93
0.22
0.69
3.25
10.36
ISO-GRID
Reference
6.60
6.67
Overallb
RSDR %
0.98
0.53
RSDr %
1.20
Yogurt
sR
1.20
1.18
Orange juice
sr
ISO-GRID
Flour/meal
Reference
ISO-GRID
Walnuts
Overall
Raw ground beef
Precision estimates
Precision estimates were calculated after conversion of the data to log10; r = 2.8 sr; R = 2.8 sR; sr = repeatability standard deviation; sR =
reproducibility standard deviation; RSDr = repeatability relative standard deviation; RSDR = reproducibility relative standard deviation.
Difference between the methods is statistically significant at the 5% level.
Diluent
Enzyme
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
PTb
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
Peptone
PT
PT
PT
None
None
Papain
Papain
None
None
None
None
None
Hemicellulase
Papain
Papain
None
None
None
None
None
None
Papain
Papain
None
None
None
Table 1. Collaborative study results for yeast and mold counts in garlic powder by test and reference methods
Results of reference method, count/g
1 (C)a
Laboratory
1
2
3
4
5
6
7
8
9
10
d
11
12
13
14
15
16
17
18
19
20
a
b
c
d
2 (A)
1.0 101b
3.3 10
1b
2.3 101b
6.7 100b
1.7 10
3.3 100b
1.0 10
1b
6.7 101b
3.3 10
0b
3.3 101b
3.3 10
0b
5.5 103
b
1.7 10
<1.0 101
b
2.7 10
1.2 104
2b
1.0 101b
1.0 10
<1.0 10
3.3 100b
<1.0 10
1.3 101b
3.3 10
1b
<1.0 10
1.3 102b
1.7 10
<1.0 10
2.3 101b
4.7 10
1b
3.3 100b
6.7 10
6.7 100b
0b
<1.0 10
<1.0 10
b
1.7 101b
6.1 10
6.7 100b
3.0 10
1.0 10
2b
3.3 100b
6.7 10
0b
0b
9.4 103
b
2.7 101b
1b
6.7 10
1.7 101b
3.3 10
1b
1.0 102b
6.7 10
0b
0b
3.3 10
8.9 103
b
2.3 101b
3.3 101b
3.3 10
0b
3.3 100b
1.0 10
3.3 10
1b
2b
1b
3.3 10
1b
6.0 103
1.7 101b
1.7 101b
<1.0 10
4.3 10
6.7 101b
1.0 10
1b
1.0 101b
3.3 100b
1b
3.3 100b
1.7 10
6.7 10
0b
3.3 100b
0b
1b
1b
3.3 10
3.3 100b
1.7 10
4.0 101b
1.0 10
1.0 101b
4.7 10
1.0 102b
1b
1b
1b
3.3 10
1.0 101b
1.0 104b
3.3 101b
b
b
1.0 10
1.0 10
1b
1.7 10
6.7 10
1.3 101b
6.7 101b
1b,c
6.7 100b
1.0 10
1 (C)
5.7 10
1b
1b
b,c
1.3 101b
1b
3.0 10
<1.0 101
3.3 100b
6 (A)
1b
2b
1b
8.3 10
1b
6.7 100b
1b
2.7 10
3.3 100b
1b
<1.0 10
2.0 101b
2.7 101b
3.3 10
5 (C)
0b
0b
6.7 10
1b
3.3 100b
b
b
1.0 10
1b
<1.0 101
3.7 10
2.3 101b
1.0 10
4 (B)
1b
<1.0 101
1b
3 (B)
3.3 10
1b
4.3 101b
<1.0 10
3.3 100b
4.7 10
1b
3.3 100b
<1.0 10
2.0 101b
3.3 10
0b
2 (A)
3 (B)
4 (B)
5 (C)
6 (A)
1.0 101
2.0 101
<1.0 101
4.0 101
<1.0 101
<1.0 101
2.0 102
6.0 10
1.0 10
7.0 10
<1.0 10
1.0 10
<1.0 101
<1.0 101
<1.0 101
<1.0 101
1.0 101
<1.0 101
2.0 10
1.0 10
<1.0 10
1.0 10
1.0 10
2.0 101
6.0 101
1.0 101
7.0 101
<1.0 10
2.0 101
<1.0 10
2.0 101
<1.0 10
1.0 101
<1.0 10
<1.0 10
1.0 101
1.0 101
<1.0 101
2.0 101
3.0 101
<1.0 101
<1.0 101
<1.0 101
<1.0 10
1.0 10
<1.0 10
<1.0 10
<1.0 10
2.0 101
2.0 101
1.0 101
2.0 101
1.0 101
2.0 101
1.0 101
8.9 103
9.4 103
3.0 10
5.8 103
<1.0 10
1.3 10
1.2 104
<1.0 10
4.0 10
1.9 104
5.0 10
1.0 10
6.0 103
<1.0 10
<1.0 10
<1.0 10
<1.0 101
<1.0 101
<1.0 101
1.0 101
1.0 101
3.0 101
5.0 101
2.6 102
1.0 101
2.0 101
<1.0 101
<1.0 101
<1.0 101
<1.0 10
<1.0 101
1.2 10
1.1 10
3.0 10
1.0 101
4.0 101
4.0 101
<1.0 10
1.0 101
<1.0 10
1.0 10
3.0 101
<1.0 10
<1.0 10
1.0 101
<1.0 10
1.1 10
c
1.0 101c
1.0 10
1.0 101
<1.0 10
2.5 10
<1.0 10
<1.0 101
<1.0 101
<1.0 101
<1.0 101
<1.0 101
1.0 101
1.0 10
<1.0 10
1.0 10
1.0 10
<1.0 10
3.0 101
Letter in parentheses refers to the lot from which the sample was drawn.
Estimated count. Counts on one or more pour plates fall outside the specified counting range of 10150 colonies/plate.
Estimated count due to spreader.
Statistical outlier for both methods. Data excluded from statistical analysis.
Table 2. Collaborative study results for yeast and mold counts in raw ground beef by test and reference methods
Results of reference method, count/g
1 (A)a
Laboratory
2 (A)
3 (B)
4 (C)
3 (B)
4 (C)
5 (C)
6 (B)
4.7 103
3.4 104
5.8 10
3.5 103
4.5 103
1.7 104
5.6 10
1.3 10
1.1 10
1.8 103
3.1 103
2.5 103
1.0 104
3.2 103
6.4 103
3.5 103
7.2 103
2.3 103
4.5 104
2.5 10
1.8 10
5.2 10
1.0 10
4.5 10
2.9 10
1.0 10
4.4 10
2.7 10
1.1 10
3.9 103
2.9 103
1.3 104
3.2 104
1.3 105
9.6 103
1.0 103
1.6 103
8.4 103
5.9 104
1.1 105
3.9 103
8.6 10
5.6 103
3.7 103
10
3.4 10
1.4 104b
11
12
1.7 10
13
5.7 103
5.4 10
3.5 103
b
1.5 10
3b
1.4 10
7.3 10
6.1 10
5.6 10
4.3 103
1.4 105
8.4 104
1.7 104
7.2 103
2.9 10
9.8 10
2.8 10
4.2 10
2.5 10
3.6 103
1.4 104
9.0 104
9.1 104
7.3 103
3.1 10
7.1 10
4.3 10
5.0 10
1.0 10
1.1 10
3.1 10
7.5 10
4.6 103
3.7 104
5.8 103
5.6 10
2.6 10
b
1.6 104b
7.6 10
1.8 104
1.0 10
4.1 104
b
2.7 10
4b
3.9 10
2.5 10
5.6 10
1.7 10
1.6 10
5.0 10
1.2 10
5b
1.7 104
1.3 104
2.7 103
1.4 104
7.9 103
8.0 103
3.2 103
7.2 103
2.1 103
4.1 10
7.2 10
2.7 10
8.7 10
2.9 10
1.1 10
1.8 104b
9.1 103
2.9 104
2.5 104
9.7 103
3.1 104
3.1 104
9.1 103
6.2 10
1.8 10
3.5 10
5.7 10
8.1 10
1.7 10
1.7 10
2.1 103
4.1 103
4.3 103
1.4 104
6.1 103
2.0 104
1.5 103
2.3 103
3.3 104
2.0 10
4.1 10
6.9 10
4.3 10
4.1 10
7.0 104
2.1 103
2.5 104
4.4 103
5.4 103
3.0 103
4.1 103
6.1 103
3.8 104
4.5 104
4.1 103
16
6.2 103
17
18
2.0 10
3.9 104
2.2 103
2.1 103
3.8 105
2.1 104
4.4 104
1.4 104
5.4 10
2.9 10
6.5 10
1.9 10
7.3 10
5.1 10
3.1 10
1.8 10
1.9 10
7.3 10
7.0 10
1.4 103
20
2.4 10
2.1 10
2.0 10
9.6 10
3.0 10
3.9 104
2.0 104
4.8 10
1.9 104
1.2 104
2.3 10
1.4 104
3.0 103
1.2 10
1.3 103
5.2 103
5.7 10
3.3 102b
19
9.7 10
N/A
2.6 103
8.0 10
N/A
4.2 103
15
5.6 10
N/A
N/A
2.4 10
N/A
6.1 104
14
3.9 10
N/A
1.1 104
3.0 103
3.8 103
2.2 103
6.7 103
2.5 104
9.6 103
8.7 10
2b
1.4 104
7.8 10
2.5 10
4.1 104b
4.1 103
1.2 104b
4
9.7 10
1.7 104
2 (A)
3.4 103
2b
4.4 104
1 (A)
5.3 10
1.5 105
6 (B)
1.4 104b
3.5 103
1.4 103
<1.0 102
<1.0 102
<1.0 102
1.4 104
2.0 103
5.9 10
9.8 10
2.6 10
3.7 10
2.4 10
1.9 10
6.2 10
1.8 104
Letter in parentheses following sample number refers to the lot from which the sample was drawn.
Estimated count. Counts on one or more pour plates fall outside the specified counting range of 10150 colonies or number of positive squares on filter exceeded 1520 (>95% of squares positive).
Statistical outlier for the reference method. Data excluded from statistical analysis.
N/A, valid counts not available for reference method. Collaborator used a single pipette to transfer all dilutions into Petri dishes and proceeded from lowest dilution to highest dilution, resulting in
carryover of significant inoculum from each dilution to the next higher. Counts on lowest dilution were too numerous to count on all 6 samples, and results of higher dilutions were compromised by
this technique error. Data excluded from statistical analysis.
Statistical outlier for test method. Data excluded from statistical analysis.
Table 3. Collaborative study results for yeast and mold counts in walnut pieces by test and reference methods
Results of reference method, count/g
1 (A)a
Laboratory
2 (A)
3 (B)
4 (B)
6 (C)
1 (A)
2 (A)
3 (B)
4 (B)
5 (C)
6 (C)
9.4 103
3.1 104
2.5 104
3.1 104
2.2 104
1.8 104
1.4 104
4.1 103
1.1 104
8.7 103
4.8 103
4.1 103
3.8 10
3.9 103
2.3 103
7.9 103
2.5 104
2.4 104
2.7 103
9.4 103
1.6 103
1.0 104
2.2 104
2.3 104
2.8 103
5.3 103
3.1 10
3.0 10
4.6 10
8.5 10
6.6 10
3.3 10
2.5 10
9.0 10
4.9 10
3.5 10
3.8 10
1.0 103
4.5 10
1.6 10
1.6 10
4.7 10
4.9 10
3.7 10
1.3 10
1.2 10
3.9 10
2.4 103
2.7 103
5.3 103
4.7 103
2.5 103
5.5 10
3.8 10
1.9 10
1.8 10
1.3 10
5.3 103
2.9 103
1.5 104
5.8 10
4.5 103
3.4 103
2.5 104
2.5 104
5.1 103
7.4 103
2.0 103
3.2 103
9.4 103
10
1.1 10
11
1.9 104
4.3 103
1.7 104
1.7 104
2.0 104
5.6 103
1.2 104
4.4 103
1.3 104
12
3.9 10
2.0 10
2.2 10
2.8 10
3.6 10
5.3 10
9.0 10
7.0 10
4.8 10
13
2.8 103
4.0 103
1.9 104
2.0 104
4.3 103
5.4 103
2.4 103
3.4 103
7.7 103
6.9 103
<1.0 10
5.3 10
5.6 10
1.5 10
2.1 10
1.8 104c
1.8 10
2.3 10
1.2 10
4c
3.4 103
3.1 103
8.7 103
8.5 103
4.6 103
6.1 103
1.4 10
3.3 10
9.7 10
7.2 10
4.1 10
4.5 103
3.2 103
5.3 103
3.4 103
3.5 103
1.3 104
2.9 104
2.8 103
6.0 103
3.0 103
1.1 104
4.3 103
7.5 103
5.7 103
1.0 104
2.2 104
5.7 103
4.6 10
1.9 10
1.2 103
4.6 103
5.9 103
4.4 10
9.3 10
1.2 10
1.8 10
2.0 10
1.4 10
9.5 10
5.6 10
1.0 102
1.3 104
4.0 103
8.6 103
6.9 103
3.5 103
1.5 104
1.3 104
3.5 103
7.7 103
16
4.1 10
8.7 103
17
4.5 103
4.7 103
1.2 104
1.6 104
4.0 103
7.0 103
2.6 103
4.3 103
4.6 103
6.4 103
2.2 103
3.2 103
18
5.5 10
4.4 10
1.1 10
1.0 10
3.4 10
1.1 10
6.6 10
4.4 10
1.0 10
1.8 10
3.7 10
1.3 104
19
20
a
b
c
d
e
5.1 103e
1.1 10
7.0 103e
1.6 104
3.0 104
5.3 103
5.5 10
1.8 10
2.4 10
6.0 10
1.5 10
1.3 10
2.7 10
1.5 104
3.3 10
1.0 10
2.7 10
4.5 10
4.0 10
2.5 10
4.6 103
6.6 10
1.0 10
3.3 10
4.3 10
6.3 10
4.8 10
4.3 103
1.7 10
5.3 10
6.2 10
15
2.1 10
1.3 10
3.0 10
3.5 10
1.3 10
14
2.8 10
4.0 103
3.1 10
1.1 10
4.7 10
1.0 104e
2.1 103
3.5 103
9.6 103
9.8 103
2.2 103
4.1 103
5.3 10
3.9 10
8.6 10
1.8 10
2.1 10
7.5 10
7.4 103
Letter in parentheses following the sample number refers to the lot from which the sample was drawn.
Statistical outlier for reference method. Data excluded from statistical analysis.
Estimated count. Counts on one or more pour plates fall outside the specified counting range of 10150 colonies.
Statistical outlier for test method. Data excluded from statistical analysis.
Estimated count due to spreader.
Table 4. Collaborative study results for yeast and mold counts in flour/meal by test and reference methods
Results of reference method, count/g
1 (B)a
Laboratory
2 (A)
3 (C)
4 (A)
8.7 102b
6 (C)
1 (B)
2 (A)
3 (C)
4 (A)
3.3 104
2.7 103
4.0 10
2.8 104
1.5 103
1.8 103
4.8 10
3.8 10
3.0 10
1.9 104
4.4 103
2.8 10
2.5 10
3.1 104
3.1 103
5.0 103
1.1 10
3.4 104
3.6 103
3.0 103
3.8 103
8.9 104
3.3 103
9.0 103
3.6 103
3.0 102
10
4.2 10
11
9.9 104
9.3 103
3.8 103
12
5.4 10
3.5 10
3.3 10
13
8.4 104
5.1 103
14
15
16
3.5 10
8.3 104
b
4.3 10
4c
3.1 10
3.4 10
4.1 10
2.7 10
3.4 103
2.8 10
18
2.1 10
3.9 10
20
a
b
c
5.0 104c
4.0 103
3.7 10
3.3 10
3.9 10
4.1 10
2b
1.6 105
1.3 103
2.2 104
1.8 103
1.2 103
3.2 103
1.7 104
2.2 103
4.9 103
3.5 10
2.4 103
b
5.7 10
2b
4.2 10
2.4 104
b
3.2 10
5b
4.0 10
2.3 103
b
1.6 10
2.3 103
4.4 104
2.6 10
3.2 10
3.2 103
6.1 104
3.2 10
4.0 10
6.0 104
b
4.0 10
2b
7.7 10
5.9 10
8.5 104
4.3 10
8.3 10
2.5 10
1.5 10
1.0 10
5.7 104
1.3 105
b,c
1.8 10
3b
4.0 103
3.4 10
5.5 10
4c
2.5 103b
2.6 103
1.0 105
3.5 10
4.4 10
3.2 10
6.3 102b
4.2 10
1.7 103c
9.6 104
2.8 10
6.3 10
1.6 103
2.5 10
2b
7.7 102b
3.4 103
2.4 10
4b
2.7 102b
3b
3.3 102
6 (C)
3.3 103
9.3 102c
1.1 10
2.2 103
19
4.3 10
1.8 104
4.0 102b
17
4.9 10
5 (B)
3b
1.0 103b
8.0 10
2b
7.0 102b,c
2.9 10
3c
4.5 10
1.6 10
4.2 10
1.5 10
3.3 10
2.6 104
1.4 103
1.6 103
2.2 103
2.4 104
2.6 103
6.0 10
3.3 10
2.8 10
4.0 10
7.5 10
3.3 104
1.1 104
1.4 103
6.0 102
2.9 103
2.4 104
8.0 102
1.8 10
1.1 10
8.0 10
2.4 10
1.7 10
1.5 103
2.6 104
2.8 103
3.3 103
3.1 103
2.1 104
3.3 103
4.8 103
2.2 103
2.0 104
1.3 103
1.4 103
5.8 10
3.2 10
2.8 10
1.9 10
1.0 10
4.0 10
3.4 10
3.1 10
7.2 10
2.3 10
6.1 104
4.2 103
3.7 103
2.3 104
6.2 104
1.6 103
1.6 10
2.2 10
1.1 10
7.0 10
2.0 10
1.0 103
4.9 104
1.7 103
2.6 103
1.8 103
5.5 104
5.6 103
6.2 103
1.9 10
6.0 10
9.0 10
1.5 10
2.9 10
1.1 105
2.3 103
5.0 103
3.4 103
7.8 104
6.5 103
4.4 103
9.1 10
6.0 10
4.3 10
1.9 10
8.6 10
4.7 102b
2.3 104
2.8 103
7.6 103
1.6 103
1.0 105
1.0 103
3.1 10
2.8 10
2.1 10
1.6 10
5.9 10
3.5 10
9.4 103
7.7 102b
1.7 105
1.8 103
1.3 103
1.8 103
6.3 104
5.1 103
2.2 10
3.3 10
1.0 10
1.0 10
1.1 10
6.3 10
3.2 103
Letter in parentheses following the sample number refers to the lot from which the sample was drawn.
Estimated count. Counts on one or more pour plates fall outside the specified counting range of 10150 colonies.
Estimated count due to spreader.
Table 5. Collaborative study results for yeast and mold counts in orange juice by test and reference methods
Results of reference method, count/mL
1 (A)a
Laboratory
2 (B)
3 (A)
1.1 103
2.5 105
1.1 10
4.2 10
4 (C)
9.0 102c
1.2 10
6 (C)
2.3 104
3.3 105
4.7 105
1.0 103
3.9 106
1.0 103
2.9 106
1.2 107
1.8 106
3.6 106
9.5 10
3.9 10
1.4 10
1.2 10
3.4 10
2.0 10
3.5 10
2.8 10
9.6 102
1.1 107
1.3 103
1.2 106
7.6 106
2.2 106
1.2 10
1.4 10
5.0 10
3.9 10
1.0 10
7.1 10
9.3 10
2.4 10
5.6 10
6.6 10
9.2 10
6.2 105
>1.5 106
5.5 105
>1.5 106
2.1 105
4.9 105
4.4 10
3.5 10
3.0 10
3.3 10
2.0 10
2.0 103
3.0 107
2.3 103
2.8 106
2.1 107
2.5 106
9.5 102
2.1 107
1.4 103
11
9.1 10
12
13
h
14
2.4 103
c
1.2 10
3c
N/A
>1.5 10
2.1 107
2.1 10
2.9 107
c
9.3 10
5c
1.2 10
1.6 103c
2.9 10
1.1 103
2.6 105
1.2 103
8.1 105
1.5 107
5.3 105
1.0 10
2.0 10
2.6 10
3.7 10
1.6 10
5.2 106
3.0 106
2.2 107
3.7 106
>1.0 108
1.7 10
>1.0 10
1.2 10
>1.0 10
>1.0 10
1.7 104
7.2 105
3.2 105
1.8 103
1.8 107
9.1 102
2.3 106
1.9 107
3.1 106
7c
2.2 106
1.2 10
3.2 10
1.1 10
6.9 10
1.9 10
2.5 10
1.2 10
5.0 10
5.1 102
2.7 105
2.6 107
2.0 104
1.5 10
1.8 10
3.5 10
1.5 10
2.1 10
3.1 10
1.6 10
2.0 10
1.1 10
1.0 105
17
2.5 10
18
3.6 103
3.4 107
2.8 103
19
1.5 10
8.1 10
1.1 10
20
3.6 103
3.4 10
1.6 103
c
1.5 10
4.8 105
3
1.2 106
2.0 107
>1.5 10
3.1 107
1.3 10
5c
1.4 103
1.8 103
2.4 105
c
2.7 105
16
3.2 10
>1.5 10
5.0 107
1.2 10
6.1 10
2.6 105
N/A
5
>1.5 10
2.1 103
N/A
3
15
5 (B)
1.2 106
4 (C)
9.1 105
1.8 103
3 (A)
1.4 106
10
2 (B)
1.3 103
1.1 10
1 (A)
1.5 106
6 (C)
1.0 103
5 (B)
3c
3.5 103
1.0 10
5c
1.3 105
c
6.0 10
4c
4.3 105
c
1.0 10
5c
1.3 105
N/A
N/A
5
1.2 106
5.4 10
c
1.3 10
7c
3.1 107
7.1 10
1.7 107
5.7 10
4c
2.9 105c
3.3 10
6.7 105
c
1.2 10
5c
7.2 105
7.3 107c
3.2 103
2.4 107
5.3 103
1.1 107
8.2 106
1.7 105
1.7 10
1.1 10
1.3 10
8.0 10
2.0 10
1.0 103
1.7 107
1.0 103
2.3 106
1.7 107
2.3 106
1.8 106
6.7 10
1.0 10
8.2 10
2.1 10
3.3 10
2.5 103
2.9 107
8.9 102
3.1 106
2.8 107
4.8 106
1.6 10
4.2 10
1.6 10
8.2 10
2.7 10
2.3 106
1.7 107
7.3 105
8.6 102
1.7 107
1.0 103
5.6 105
Letter in parentheses following the sample number refers to the lot from which the sample was drawn.
Statistical outlier for reference method. Data excluded from statistical analysis. Collaborator also reported a scheduling deviation. Samples were initiated on December 13, 1994, instead of
December 7, 1994.
Estimated count. Counts on one or more pour plates fall outside the specified counting range of 10150 colonies or number of positive squares on filter exceeded 1520 (>95% of squares positive).
Statistical outlier for both methods. Data excluded from statistical analysis.
Statistical outlier for test method. Data excluded from statistical analysis.
Samples 1 and 3 are statistical outliers for reference method. Data from these 2 samples excluded from statistical analysis.
Samples 2, 4, 5, and 6 are indeterminately high for both methods. Data from these 4 samples excluded from statistical analysis.
N/A, valid counts not available for reference method. Collaborator used a single pipette to transfer all dilutions into Petri dishes and proceeded from lowest dilution to highest dilution, resulting in
carryover of significant inoculum from each dilution to the next higher. Counts on lowest dilution were too numerous to count on 5 of 6 samples, and results of higher dilutions were compromised by
this technique error. Data excluded from statistical analysis.
Table 6. Collaborative study results for yeast and mold counts in yogurt by test and reference methods a
Results of reference method, count/g
1 (A)b
Laboratory
c
2.6 10
7d
2.2 106
d
1.8 10
7d
2.5 107d
2.3 10
7d
>1.5 107
9.9 106
1.2 107
3.7 10
9.7 10
2.0 10
5.0 10
1.2 107
d
1.8 107d
>1.5 10
2.1 10
12
13
16
17
18
19
20
5.3 106
d
2.1 10
>1.5 10
1.6 107d
1.5 10
7d
1.6 107d
1.8 10
3.3 10
1.6 107d
2.2 10
7d
7d
2.0 107d
1.9 10
1.5 10
1.3 107
>1.5 10
1.6 107d
2.6 10
9.9 106
9.5 10
5.5 105
2.7 10
>1.5 10
1.6 107d
1.2 10
7d
9.8 106
2.2 10
1.3 107
2.2 10
1.4 10
2.3 107d
1.5 107d
2.4 10
1 (A)
7.7 106
2.2 10
7d
2 (B)
>1.0 107
d
8.0 10
6d
3 (C)
>1.0 107
1.2 106
>1.0 107
>1.0 107
>1.0 10
>1.0 107
2.2 10
1.1 10
2.8 10
8.8 10
7.4 107d
1.4 10
2.0 107d
>1.5 10
1.8 107d
1.7 10
1.7 106
d
3.2 10
7d
4.4 106
2.3 10
1.3 10
1.4 107
>1.5 10
2.1 107d
1.8 107d
2.7 10
7d
1.8 107d
2.2 107d
1.9 10
7d
1.3 107
7.5 106d
2.4 10
9.6 106d
1.7 105
>1.0 107
>1.0 10
>1.0 10
>1.0 107
7.4 10
5.8 10
6d
9.6 10
6d
>1.0 107
>1.0 107
>1.0 107
>1.0 107
>1.0 107
>1.0 107
>1.0 107
>1.0 10
2.2 10
>1.0 10
2.4 107d
1.1 10
1.6 10
6.0 10
>1.0 107
7d
7.8 106d
5.9 105
7.9 106
>1.0 107
>1.0 10
1.1 10
2.9 10
>1.0 107
6d
>1.0 107
8.5 106d
>1.0 10
>1.0 10
>1.0 107
>1.0 107
1.4 10
>1.0 10
>1.0 107
1.3 107
7d
1.5 10
N/A
7
7d
6 (C)
1.5 106
>1.0 107
5 (B)
1.0 107
4 (A)
>1.0 107
1.7 107d
7d
2.9 10
7d
N/A
7d
1.7 107d
7d
1.7 10
7d
6 (C)
4.3 106
d
N/A
1.4 107
d
3.4 106
1.3 10
6.8 10
7d
2.4 107d
1.1 10
1.3 107d
N/A
7d
1.8 107d
1.3 10
7.3 106
N/A
1.4 10
1.4 106
N/A
d
2.8 10
7d
1.7 107d
7d
15
2.1 107d
2.0 10
7d
1.7 107d
11
14
10
3.5 106
d
>1.5 107
5 (B)
2.6 107d
4 (A)
4.9 10
3 (C)
2 (B)
5.4 106d
7
1.8 10
2.9 106
>1.0 10
>1.0 10
>1.0 107
d
9.2 10
6d
4.7 106
>1.0 10
>1.0 10
>1.0 10
8.9 106d
>1.0 107
>1.0 107
1.4 106
3.2 106
3.5 106
1.2 10
3.1 106
>1.0 10
1.7 10
>1.0 10
>1.0 107
2.9 105
1.9 105
1.5 105
3.4 105
1.0 106
1.7 105
>1.0 107
>1.0 107
>1.0 107
>1.0 107
>1.0 10
>1.0 10
>1.0 107
>1.0 107
8.3 10
6d
8.0 10
6d
>1.0 107
>1.0 107
>1.0 10
4.5 10
6.4 106d
>1.0 107
>1.0 10
7.7 106
d
8.9 10
6d
>1.0 10
9.6 10
6d
>1.0 10
8.5 10
6d
9.2 106d
>1.0 107
>1.0 107
>1.0 107
>1.0 107
9.6 10
6d
>1.0 107
8.5 10
6d
4.3 106
9.2 10
6d
>1.0 107
7.6 106d
8.7 106d
Data set contains many samples for which the test method was indeterminately high (i.e., >1.0 107/g) and for which the reference gave either an indeterminately high value or else gave a
determinately high result in excess of 1.0 107/g. All such samples were excluded from statistical analysis.
Letter in parentheses following the sample number refers to the lot from which the sample was drawn.
Scheduling deviation. Samples initiated on December 13, 1994, instead of December 7, 1994. Data excluded from statistical analysis.
Estimated count. Counts on one or more pour plates fall outside the specified counting range of 10150 colonies or number of positive squares on filter exceeded 1520 (>95% of squares positive).
N/A, valid counts not available for reference method from this collaborator. Collaborator used a single pipette to transfer all dilutions into Petri dishes and proceeded from lowest dilution to highest
dilution, resulting in carryover of significant inoculum from each dilution to the next higher. Counts on the lowest dilution were too numerous to count on all 6 samples, and results of higher dilutions
were compromised by this technique error. Data excluded from statistical analysis.